FRESHWATER BIOLOGY AND
POLLUTION ECOLOGY
TRAINING MANUAL
U.S. ENVIRONMENTAL PROTECTION AGENCY
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FRESHWATER BIOLOGY
This course is designed as an introduction to aquatic
biolog} for samtary engineei s, scientists, and othei s
who are invoked in freshwater pollution studies,
surveillance, and i_onttol Biologists new to (ho field
of aquatic biology and pollution problems may find it
useful foi orientation.
AND
FRESHWATER POLLUTION ECOLOGY
This course is oliered lor aquatic biologists 01 persons
with comparable experience concerned with and/oi
involved in the application of biological principles,
techniques and parameters to pollution studies and
abatement piograms
ENVIRONMENTAL PROTECTION AGENCY
Water Programs Operations
TRAINING PROGRAM
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TRAINING PROGRAM
Through the Water Programs Operations Office, Environmental
Protection Agency conducts programs of research, technical
assistance, enforcement, and technical training for water
pollution control.
The objectives of the Training Program are to provide specialized
training in the field of water pollution control which will lead to
rapid application of new research findings through updating of
skills of technical and professional personnel, and to train
new employees recruited from other professional or technical
areas in the special skills required. Increasing attention is
being given to development of special courses providing an
overview of the nature, causes, prevention, and control of
water pollution.
Scientists, engineers, and recognized authorities from other
Agency programs, from other government agencies, universities,
and industry supplement the training staff by serving as guest
lecturers. Most training is conducted in the form of short-term
courses of one or two weeks' duration. Subject matter includes
selected practical features of plant operation and design, and
water quality evaluation in field and laboratory. Specialized
aspects and recent developments of sanitary engineering, chemistry,
aquatic biology, microbiology, and field and laboratory techniques
not generally available elsewhere, are included.
The primary role and the responsibility of the states in the
training of wastewater treatment plant operators are recognized.
Technical support of operator-training programs of the states is
available through technical consultations m the planning and
development of operator-training courses. Guest appearances
of instructors from the Environmental Protecticn Agency, and
the loan of instructional materials such as lesson plans and
visual training aids, may be available through special arrangement.
These training aids, including reference training manuals, may
be reproduced freely by the states for their own training programs.
Special categories of training for personnel engaged in treatment
plant operations may be developed and made available to the states
for their own further production and presentation.
A bulletin of courses is prepared and distributed periodically
by the National Training Center. The bulletin includes descriptions
of courses, schedules, application blanks, and other appropriate
information. Organizations and interested individuals not on
the mailing list should request a copy from The National Training
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FOREWORD
These manuals are prepared for reference use of students enrolled in
scheduled training courses of the Office of Water Programs, Environmental
Protection Agency.
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COURSE DESCRIPTION
FRESHWATER BIOLOGY AND
POLLUTION ECOLOGY (140)
2 weeks
CINCINNATI, OHIO June 4-15, 1973
This course is designed for technical
personnel (other than biologists) engaged
in water quality analysis and management
Within the framework of his personal
background capacities, and experience, the
student should be able, on completion of
this course to
Understand many basic environmental
factors impinging on aquatic
communities
Recognize or identify to broad groups
most freshwater organisms
commonly encountered, using
correct procedures and appropriate
literature when available and also
using judgment in assesing his own
technical capacity in regard to the
degree of identification attempted
Select and use appropriate common types
of biological field collection equip-
ment and procedures
Select and use appropriate types of
biological laboratory analytical
equipment and procedures
Perform simple analyses of an aquatic
community in order to assess the
likelihood that it may have been
disturbed by pollution
Recognize gross biological indications
of particular types of pollution
when present
Predict possible effects of a given type
of pollutant on a given habitat
Organize a field survey to determine
the severity and extent of pollution.
Course work includes lectures, discussions,
problem assignments, and laboratory sessions.
Field work is included to allow student partici-
pation in selecting and using biological field
collection equipment and familiarization with
biological communities.
Representative topics usually include:
Types of aquatic organisms
Aquatic organisms or significance
m water quality
Lake, reservoir, and stream sampling
techniques
Use of artificial substrates
Thermal pollution
Investigation of fish kills
Environmental quality
Eutrophication in the freshwater
environment
Biological magnification
Participants should bring appropriate
clothing for field work, including rainwear.
Boots will be supplied locally unless notice
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U.S. Environmental Protection Agency
OFFICE OF WATER PROGRAMS
MANPOWER DEVELOPMENT STAFF
R. F. Guay, Director
Academic Training Branch
State and Local Operator Training
Programs
Office of Environmental Activities
Direct Technical Training Branch
National Training Center
Cincinnati, OH 45268
REGIONAL MANPOWER OFFICES
REGION I
Manpower Development Branch
Division of Air and Water Programs
424 Trapelo Road
Waltham, MA 02514
REGION VI
Manpower Development Branch
Air and Water Programs Division
1600 Patterson
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REGION II REGION VII
Manpower Development and Training Office Manpower Development Branch
Air and Water Programs Air and Water Programs
26 Federal Plaza 1735 Baltimore
New York, NY 10007 Kansas City, MO 64108
REGION III
Manpower Development Office
Air and Water Programs
Curtis Building
6th and Walnut Streets
Philadelphia, PA 19106
REGION IV
Manpower Development Branch
Division of Air and Water Programs
1421 Peachtree Street, NE, Fourth Floor
Atlanta, GA 30309
REGION V
Manpower Development Branch
Office of Air and Water Programs
1 N, Wacker Drive
Chicago, IL 60606
REGION VIU
Manpower Development Branch
Air and Water Division
1860 Lincoln Street - 9th Floor
Denver, CO 80203
REGION IX
Manpower Development Branch
Air and Water Division
100 California Street
San Francisco, CA 94111
REGION X
Manpower and Training Branch
Division of Air and Water Programs
1200 Sixth Avenue - Mail Stop 345
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CONTENTS
Title or Description Outline Number
BASIC FRESHWATER BIOLOGY
A INTRODUCTION
The Aquatic Environment 1
Significance of "Limiting Factors" to Population Variation 2
INTRODUCTION TO CLASSIFICATION OF THE BIOTA
The System of Biological Classification 3
Aquatic Organisms of Significance in Pollution Surveys 4
B PLANTS- PRODUCERS
Types of Algae 5
Blue-Green Algae 6
Green and Other Pigmented Flagellates 7
Filamentous Green Algae 8
Coccoid Green Algae 9
Diatoms 10
Key to Algae of Importance in Water Pollution 11
Aquatic Macrophytes 12
An Artificial Key to Some Common Plants 13
(Freshwater, Estuanne, and Marine)
C ANIMALS: CONSUMERS
Key to Selected Groups of Freshwater Animals 14
Biota of Wastewater Treatment Plants (Microscopic Invertebrates) 15
Biology of Zooplankton Communities 16
Macro Invertebrates 17
Aquatic Insects 18
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2
CONTENTS
Title or Description Outline Number
Freshwater Crustacea 19
Freshwater Mollusca 20
Fishes 21
Classification of Fishes 22
D BACTERIA AND FUNGI- REDUCERS
Biological Reducers (The Role of Fungi in the 23
Biodegradation of Organic Matter)
Bacteriological Indicators of Water Pollution 24
Fungi and the "Sewage Fungus" Community 25
FRESHWATER POLLUTION ECOLOGY
E WATER QUALITY AFFECTS AQUATIC COMMUNITIES
Biological Aspects of Natural Self Purification 26
Ecology of Waste Stabilization Processes 26
Effects of Pollution on Fish 27
The Interpretation of Biological Data with Reference to 28
Water Quality
F WATER QUALITY AND AQUATIC LIFE
Section III - Fish, Other Aquatic Life, and Wildlife 29
Research Needs - Fish, Other Aquatic Life, and Wildlife 30
G SOME CURRENT POLLUTION PROBLEMS
Effects of Pollution on Aquatic Life 31
The Effects of Organisms on Pollution and the Environment 31
The Effects of Pollution on Lakes 32
Water Temperature and Water Quality 33
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CONTENTS
3
Title or Description Outline Number
Biotic Effects of Solids 35
Global Deterioration and our Ecological Crisis 36
H BIOLOGICAL METHODS AND TECHNIQUES
Fundamentals of the Toxicity Bioassay 37
Biomonitormg of Industrial Effluents 40
Biological Field Methods 41
Stream Invertebrate Drift 42
Artificial Substrates 43
Attached Growths (Periphyton or Aufwuchs) 44
Application of Biological Data 46
Procedures for Fish Kill Investigations 47
An Initiation into Statistics 48
Using Benthic Biota in Water Quality Evaluations 49
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SURVEY OF THE BIOTA
The first half of this manual is a synoptic review of the components
of the aquatic community to aid the observer in recognizing many
common types of organisms encountered both in the field, laboratory,
and treatment facilities.
A comprehension of the system of biological nomenclature and
ecological classifications, is basic to an understanding of aquatic
life communities. The first exercise in ecology is systematics.
Application of systematics will depend on one's background, present
limitations, and program objectives.
SystematicB, broadly defined, is the study of the diversity of organisms.
In connotation it is a wedding between taxonomy and ecology. In applied
biology or ecology, good systematics is indispensable and is an immensely
useful system of information storage and retrieval. The following
definitions are basic.
SYSTEMATICS: "The scientific study of the kinds and diversity of
organisms and of any and all relationships among them. "
CLASSIFICATION: "The ordering of organisms into groups (or sets)
on the basis of their relationships, that is, of their associations by
contiguity, similarity, or both."
TAXONOMY: "The theoretical study of classification, including its
bases, principles, procedures, and rules."
IDENTIFICATION: "The use of a key (or key substitute like an expert)
to place an unknown organism into a specific taxonomic rank. "
Section A INTRODUCTION
Section B PLANTS: PRODUCERS
Section C ANIMALS: CONSUMERS
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PAGE NOT
AVAILABLE
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THE AQUATIC ENVIRONMENT
Part 1: The Nature and Behavior of Water
I INTRODUCTION
The earth is physically divisible into the
lithosphere or land masses, and the
hydrosphere which includes the oceans,
lakes, streams, and subterranean waters,
A Upon the hydrosphere are based a number
of sciences which represent different
approaches. Hydrology is the general
science of water itself with its various
special fields such as hydrography,
hydraulics, etc. These in turn merge
into physical chemistry and chemistry.
B Limnology and oceanography combine
aspects of all of "these, and deal not only
with the physical liquid water and its
various naturally occurring solutions and
forms, but also with living organisms
and the infinite interactions that occur
between them and their environment.
C Water quality management, including
pollution control, thus looks to all
branches of aquatic science in efforts
to cooi dinate and improve man's
relationship with his aquatic enwronmcnt.
II SOME FA CTb A BOUT \VA TErt
A Wateti^is the only abundant liquid on our
plan_et. It has many pi operties most
unusual for liquids, upon which depend
most of the iamiliar aspects ot the world
about us Jb we know it.
TABLE 1
UNIQUE PROPERTIES OF WATER
Property
Significance
Highest heat capacity (specific heat) of any
solid or liquid (except NH^)
Stabilizes temperatures of organisms and
geographical regions
Highest latent heat of fusion (except NH )
Thermostatic effect at freezing point
Highest heat of evaporation of any substance
Important in heat and water transfer of
atmosphere
The only substance that has its maximum
density as a liquid (40C)
Fresh and brackish waters have maximum
density above freezing point. This is
important in vertical circulation pattern
in lakes.
Highest surface tension of any liquid
Controls surface and drop phenomena,
important in cellular physiology
Dissolves more substances in greater
quantity than any other liquid
Makes complex biological system possible.
Important for transportation of materials
in solution.
Pure water has the highest di-electric
constant of any liquid
Leads to high dissociation of inorganic
substances in solution
Very little electrolytic dissociation
Neutral, yet contains both H+ and OH ions
Relatively transparent
Absorbs much energy in infra red and ultra
violet ranges, but little in visible range.
Hence "colorless"
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The Aquatic Environment
B Physical Factors of Significance
1 Water substance
Water is not simply "HgO" but in
reality is a mixture of some 33
different substances involving three
isotopes each of hydrogen and oxygen
(ordinary hydrogen H^", deuterium H ,
and tritium H^, ordinary oxygen O*®,
oxygen 17, and oxygen 18) plus 15 known
types of ions. The molecules of a
water mass tend to associate themselves
as polymers rather than to remain as
discrete units. (See Figure 1)
2 Density
a Temperature and density Ice.
Water is the only known substance
in which the solid state will float
on the liquid state. (See Table 2)
SUBSTANCE OF WATER
0 0 0
0 0 0
Figure 1
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The Aquatic Environment
TABLE 2
EFFECTS OF TEMPERATURE ON DENSITY
OF PURE WATER AND ICE*
Temperature (°C) Density
Water Ice **
-10
.99815
.9397
- 8
99869
. 9360
- 6
.99912
.9020
- 4
.99945
.9277
- 2
.99970
.9229
0
.99987
.9168
2
.99997
4
1.00000
6
.00997
8
.00988
10
.00973
* Tabular values for density, etc., represent
statistical estimates by various workers
rather than absolute values, due to the
variability of water.
** Regular ice is known as "ice I". Four or
more other "forms" of ice are known to
exist (ice II, ice EI, etc.), having densities
at 1 atm. pressure ranging from 1.1595
to 1.67. These are of extremely restricted
occurrence and may be ignored m most
routine operations.
This ensures that ice usually
forms on top of a body of water
and tends to insulate the remain-
ing water mass from further loss
of heat. Did ice sink, there
could be little or no carryover of
aquatic life from season to season
in the higher latitudes. Frazil or
needle ice forms colloidally at a
few thousandths of a degree
below 0° C. It is adhesive and
may build up on submerged objects
as "anchor ice", but it is still
typical ice.
1) Seasonal increase m solar
radiation annually warms
surface waters in summer
while other factors result in
winter cooling. The density
differences resulting estab-
lish two classic layers: the
epilimnion or surface layer,
and the hypolimnion or lower
layer, and in between is the
thermoclme or shear-plane.
2) While for certain theoretical
purposes a thermoclme is
defined as a zone in which the
temperature changes one
degree centigrade for each
meter of depth, in practice,
any transitional layer between
two relatively stable masses
of water of different temper-
atures (and probably other
qualities too) may be regarded
as a thermoclme.
3) Obviously the greater the
temperature differences
between epilimnion and
hypolimnion and the sharper
the gradient in the thermoclme,
the more stable will the
situation be.
4) From information given above,
it should be evident that while
the temperature of the
hypolimnion rarely drops
much below 4° C, the
epilimnion may range from
0o C upward.
5) It should also be emphasized
that when epilimnion and
hypolimnion achieve the same
temperature, stratification no
longer exists, and the entire
body of water behaves
hydrologically as a unit, and
tends to assume uniform
cnemical and physical
characteristics. Such periods
are called overturns and
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The Aquatic Environment
usually result in considerable
water quality changes of
physical, chemical, and
biological significance.
6) When stratification is present,
however, each layer behaves
relatively independently, and
considerable quality differences
may develop.
7) Thermal stratification as
described above has no
reference to the size of the
water mass, it is found in
oceans and puddles
8) The relative densities of the
various isotopes of water also
influence its molecular com-
position For example, the
lighter 016 tends to go off
first in the process of
evaporation, leading to the
relative enrichment of air by
O^g and the enrichment of
water by and Ojg. This
can lead to a measurably
higher content in warmer
climates Also, the temper-
ature of water in past geologic
ages can be closely estimated
from the ratio of in the
carbonate of mollusc snells.
b Dissolved and/or suspended solids
may also affect the density of
natural waters
TABLE 3
EFFECTS OF DISSOLVED SOLIDS
ON DENSITY
Dissolved Solids
Density
(Grams per liter)
(at 40 C)
0
1.00000
1
1.00085
2
1.00169
3
1.00251
10
1.00818
35 (mean for sea water) 1. 02822
c Density caused stratification
1) Density differences produce
stratification which may be
permanent, transient, or
seasonal.
2) Permanent stratification
exists for example where
there is a heavy mass of
brine in the deeper areas of
a basin which does not respond
to seasonal or other changing
conditions.
3) Transient stratification may
occur with the recurrent
influx of tidal water in an
estuary for example, or the
occasional influx of cold
muddy water into a deep lake
or reservoir.
4) Seasonal stratification involves
the annual establishment of
the epilimnion, hypolimnion,
and thermocline as described
above. The spring and fall
overturns of such waters
materially affect biological
productivity.
5) Density stratification is not
limited to two-layered systems,
three, four, or even more
layers may be encountered in
larger bodies of water.
d A "plunge line" may develop at
the mouth of a stream. Heavier
water flowing into a lake or
reservoir plunges below the
lighter water mass of the epiliminium
to flow along at a lower level. Such
a line is usually marked by an
accumulation of floating debris.
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The Aquatic Environment
The viscosity of water is greater at
lower temperatures (see Table 4).
This is important not only m situations
involving the control of flowing water
as in a sand filter, but also since
overcoming resistance to flow gen-
erates heat, it is significant in the
heating of water by internal friction
from wave and current action.
Living organisms more easily support
themselves in the more viscous
(and also denser) cold waters of the
arctic than in the less viscous warm
tropical waters.
TABLE 4
VISCOSITY OF WATER (In millipoises at 1 atm)
Temp. 0 C
Dissolved solids in g/ L
0
5
10
30
-10
26.0
- 5
21.4
0
17.94
18. 1
18.24
18.7
5
15. 19
15. 3
15.5
16.0
10
13. 10
13.2
13.4
13. 8
30
8.00
8. 1
8.2
8.6
100
2.84
Surface tension has biological as well
as physical significance Organisms
whose body surfaces cannot be wet by
water can either ride on the surface filrr
or m some instances may be "trapped"
on the surface film and be unable to
re-enter the water
4 Incident solar radiation is the prime
source of energy for virtually all
organic and most inorganic processes
on earth. For the earth as a whole,
the total amount (of energy) received
annually must exactly balance that
lost by reflection and radiation into
space if climatic and related con-
ditions are to remain relatively
constant over geologic time.
a For a given body of water,
immediate sources of energy
include in addition to solar
irradiation: terrestrial heat,
transformation of kinetic energy
(wave and current action) to heat,
chemical and biochemical
reactions, convection from the
atmosphere, and condensation of
water vapor.
b The proportion of light reflected
depends on the angle of incidence,
the temperature, color, and other
qualities of the water. In general,
as the depth increases arithmet-
lcally, the light tends to decrease
geometrically. Blues, greens,
and yellows tend to penetrate most
deeply while ultra violet, violets,
and orange-reds are most quickly
absorbed. On the order of 90%
of the total illumination which
penetrates the surface film is
absorbed in the first 10 meters of
even the clearest water, thus
tending to warm the upper layers.
5 Water movements
a Waves or rhythmic movement
The best known are traveling
waves caused by wind. These are
effective only against objects near
the surface. They have little
effect on the movement of large
masses of water.
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The Aquatic Environment
Standing waves or seiches occur
in lakes, estuaries, and other
enclosed bodies of water, but are
seldom large enough to be
observed. An "internal wave or
seich" is an oscillation in a
submersed mass of water such
as a hypolimmon, accompanied
by compensating oscillation in the
overlying water such that no
significant change in surface level
is detected. Shifts in submerged
water masses of this type can have
severe effects on the biota and
also on human water uses where
withdrawals are confined to a given
depth. Descriptions and analyses
of many other types and sub-types
of waves and wave-like movements
may be found in the literature.
b Tides
Tides are the longest waves known
in the ocean, and are evident along
the coast by the rhythmic rise and
fall of the water. While part and
parcel of the same phenomenon, it
is often convenient to refer to the
rise and fall of the water level as
"tide", and to the accompanying
currents as "tidal currents"
Tides are basically caused by the
attraction of the sun and moon on
water masses, large and small,
however, it is only in the oceans
and certain of the larger lakes that
true tidal action has been demonstrated.
The patterns of tidal action are
enormously complicated by local
topography, interaction with seiches,
and other factors. The literature
on tides is very large.
c Currents (except tidal currents)
are steady a rhythmic water
movements which have had major
study only in oceanography although
they are best known from rivers
and streams. They primarily are
concerned with the translocation of
water masses. They may be
generated internally by virtue of
density changes, or externally by
wind or terrestrial topography.
Turbulence phenomena or eddy
currents are largely responsible for
lateral mixing in a current. These
are of far more importance in the
economy of a body of water than
mere laminar flow.
d Coriolis force is a result of inter-
action between the rotation of the
earth, and the movement of masses
or bodies on the earth. The net
result is a slight tendency for moving
objects to veer to the right in the
northern hemisphere, and to the
left in the southern hemisphere.
While the result in fresh waters is
usually negligible, it may be con-
siderable in marine waters. For
example, other factors permitting,
there is a tendency in estuarips for
fresh waters to move toward the
ocean faster along the right bank,
while salt tidal waters tend to
intrude farther inland along the
left bank. Effects are even more
dramatic in the open oceans.
e Langmuir circulation (or L. spirals)
is the interlocking rotation of
somewhat cylindrical masses of
surface water under the influence
of wind action. The axes of the
cylinders are parallel to the
direction of the wind.
To somewhat oversimplify the
concept, a series of adjoining cells
might be thought of as chains of
interlocking gears in which at every
other contact the teeth are rising
while at the alternate contacts, they
are sinking (Figure 2).
The result is elongated masses of
waste rising or sinking together.
This produces the familiar "wind
rows" of foam, flotsam and jetsam,
or plankton often seen streaking
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The Aquatic Environment
windblown lakes or oceans. Certain
zoo-plankton struggling to maintain
a position near the surface often
collect in the down current between
two Langmuir cells, causing such
an area to be called the "red dance",
while the clear upwelling water
between is the "blue dance".
This phenomenon may be important
in water or plankton sampling on
a windy day.
6 The pH of pure water has been deter-
mined between 5. 7 and 7. 01 by various
workers. The latter value is most
widely accepted at the present time.
Natural waters of course vary widely
according to circumstances.
C The elements of hydrology mentioned
above represent a selection of some of
the more conspicuous physical factors
involved in working with water quality
Other items not specifically mentioned
include- molecular structure of waters,
interaction of water and radiation,
internal pressure, acoustical charac-
teristics, pressure-volume-temperature
relationships, refractivity, luminescence,
color, dielectrical characteristics and
phenomena, solubility, action and inter-
actions of gases, liquids and solids,
water vapor, ices, phenomena of
hydrostatics and hydrodynamics in general.
REFERENCES
1 Buswell, A.M. and Rodebush, W.H.
Water. Sci. Am. April 1956.
2 Dorsey, N. Ernest. Properties of
Ordinary Water - Substance.
Reinhold Publ. Corp. New York,
pp. 1-673. 1940.
3 Hutcheson, George E. A Treatise on
Limnology. John Wiley Company.
1957.
This outline was prepared by H. W. Jackson,
Chief Biologist, National Training Center,
Water Programs Operations, EPA, Cincinnati,
OH 45268.
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THE AQUATIC ENVIRONMENT
Part 2 The Aquatic Environment as an Ecosystem
I INTRODUCTION
Part 1 introduced the lithosphere and the
hydrosphere. Part 2 will deal with certain
general aspects of the biosphere, or the
sphere of life on this earth, which photo-
graphs from space have shown is a finite
globe in infinite space.
This is the habitat of man and the other
organisms. His relationships with the
aquatic biosphere are our common concern,
II THE BIOLOGICAL NATURE OF THE
WORLD WE LIVE IN
A We can only imagine what this world
must have been like before there was life.
B The world as we know it is largely shaped
by the forces of life.
Ill ECOLOGY IS THE STUDY OF THE
INTERRELATIONSHIPS BETWEEN
ORGANISMS, AND BETWEEN ORGA-
NISMS AND THEIR ENVIRONMENT.
A The ecosystem is the basic functional
unit of ecology. Any area of nature that
includes living organisms and nonliving
substances interacting to produce an
exchange of materials between the living
and nonliving parts constitutes and
ecosystem. (Odum, 1959)
1 From a structural standpoint, it is
convenient to recognize four
constituents as composing an
ecosystem (Figure 1).
a Abiotic NUTRIENT NUMERALS
which are the physical stuff of
which living protoplasm will be
synthesized.
1 Primitive forms of life created organic
matter and established soil.
2 Plants cover the lands and enormously
influence the forces of erosion.
3 The nature and rate of erosion affect
the redistribution of materials
(and mass) on the surface of the
earth (topographic changes).
4 Organisms tie up vast quantities of
certain chemicals, such as carbon
and oxygen.
5 Respiration of plants and animals
releases carbon dioxide to the
atmosphere in influential quantities.
6 COg affects the heat transmission of
the atmosphere.
C Organisms respond to and in turn affect
their environment. Man is one of the
most influential.
b Autotrophic (self-nourishing) or
PRODUCER organisms. These
are largely the green plants
(holophytes), but other minor
groups must also be included
(See Figure 2). They assimilate
the nutrient minerals, by the use
of considerable energy, combine
them into living organic substance.
c Heterotrophic (other-nourishing)
CONSUMERS (holozoic), are chiefly
the animals. They ingest (or eat)
and digest organic matter, releasing
considerable energy in the process.
d Heterotrophic REDUCERS are chiefly
bacterial and fungi that return
complex organic compounds back to
the original abiotic mineral condition,
thereby releasing the remaining
chemical energy.
2 From a functional standpoint, an
ecosystem has two parts (Figure 2)
1-9
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r
The Aquatic Environment
w CO NSUMERS
/>S\
PRO 0 U CERS
REDUCERS
NUTRIENT
MINERALS
FIGURE 1
a The autotrophic or producer
organisms, which construct
organic substance.
b The heterotrophic or consumer and
reducer organisms which destroy
organic substance.
3 Unless the autotrophic and hetero-
trophic phases of the cycle approximate
a dynamic equilibrium, the ecosystem
and the environment will change.
B Each of these groups includes simple,
single-celled representatives, persisting
at lower levels on the evolutionary stems
of the higher organisms. (Figure 2)
1 These groups span the gaps between the
higher kingdoms with a multitude of
transitional forms. They are collectively
called the PROTISTA.
2 Within the protista, two principal sub-
groups can be defined on the basis of
relative complexity of structure.
a The bacteria and blue-green algae,
lacking a nuclear membrane may
be considered as the lower protista
(or Monera).
b The single-celled algae and
protozoa are best referred to as
the higher protista.
C Distributed throughout these groups will
be found most of the traditional "phyla"
of classic biology.
IV FUNCTIONING OF THE ECOSYSTEM
A A food chain is the transfer of food energy
from plants through a series of organisms
with repeated eating and being eaten.
Food chains are not isolated sequences but
are interconnected.
-------
The Aquatic Environment
RELATIONSHIPS BETWEEN FREE LIVING AQUATIC ORGANISMS
Energy Flows from Left to Right, General Evolutionary Sequence is Upward
PRODUCERS 1
1 CONSUMERS
REDUCERS
Organic Material Produced,
' Organic Material Ingested or
¦ Consumed
1 Digested Internally
Organic Material Reduced
by Extracellular Digestion
and Intracellular Metabolism
to Mineral Condition
ENERGY STORED
ENERGY RELEASED
ENERGY RELEASED
Flowering Plants and
Gymnoaperms
Arachnids Mammals
Insects Birds
Basidiomycetes
Club Mosses, Ferns
Crustaceans Reptiles
Liverworts, Mosses
Segmented Worms Amphibians
Molluscs Fishes
Fungi Imperfecti
Multicellular Green
Algae
Bryozoa Primitive
Chordates
Rotifers
Roundworms Echinoderms
Ascomycetes
Red Algae
Flatworms
Brown Algae
Coelenterates
Sponges
Higher Phycomycetes
DEVELOPMENT OF MULTICELLULAR OR COENOCYTIC STRUCTURE
H 1 G
HER P R 0 T 1
Protozoa
S T A
Unicellular Green Algae
Diatoms
Pigmented flagellates
Amoeboid Cilliated
Flagellated, Suctoria
(non-pigmented)
Lower
Phycomycetes
(Chytridiales, et al )
DEVELOPMENT OF A NUCLEAR MEMBRANE
LOWER PROTISTA
Blue Green Algae
Phototropic Bacteria
(or M o n e r a )
I I
I I
I I
I I
Chemoti opic Bacteria | |
I I
Actinomycetes
Spir ochaetes
Saprophytic
Bacterial
Types
FIGURE 2
BI ECO pi 2a 1 69
-------
The Aquatic Environment
B A food web is the interlocking pattern of
food chains in an ecosystem. (Figures 3, 4)
In complex natural communities, organisms
whose food is obtained by the same number
of steps are said to belong to the same
trophic (feeding) level.
C Trophic Levels
1 First - Green plants (producers)
(Figure 5) fix biochemical energy and
synthesize basic organic substances.
2 Second - Plant eating animals (herbivores)
depend on the producer organisms for
food.
3 Third - Primary carnivores, animals
which feed on herbivores.
4 Fourth - Secondary carnivores feed on
primary carnivores.
5 Last - Ultimate carnivores are the last
or ultimate level of consumers.
D Total Assimilation
The amount of energy which flows through
a trophic level is distributed between the
production of biomass and the demands of
respiration in a ratio of approximately
1:10.
E Trophic Structure of the Ecosystem
The interaction of the food chain
phenomena (with energy loss at each
transfer) results in various communities
having definite trophic structure or energy
levels. Trophic structure maybe
measured and described either in terms
of the standing crop per unit area or in
terms of energy fixed per unit area per
unit time at successive trophic levels.
Trophic structure and function can be
shown graphically by means of ecological
pyramids (Figure 5).
Figure 3. Diagram of the pond ecosystem. Basic units are as follows: I, abiotic substances—basic inorganic and
organic compounds; IIA, producers-rooted vegetation; IIB, producers-phytoplankton; III-1A, primary consumers
(herbivores)—bottom forms; Ill-IB, primary consumers (herbivores)— zooplankton; III-2, secondary consumers (car-
nivores); III-3, tertiary consumers (secondary carnivores); IV, decomposers-bacteria and fungi of decay.
1-12
I ,u" I
I (HCMt I
-------
The Aquatic Environment
Nutrient
supply
Phytoplankton
Zooplankton
Predatory
animals
Death and decay
Bacterial
action
Mollusks
Worms
Figure 4. A MARINE ECOSYSTEM (After Clark, 1954 and Patten, 1966)
-------
The Aquatic Environment
Figure 5. HYPOTHETICAL PYRAMIDS of
(a) Numbers of individuals, (b) Biomass, and
(c) Energy (Shading Indicates Energy Loss).
Decomposers
(a)
1
Carnivores (Secondary)
Carnivores (Primary
Herbivoi es
1_
Producers
(b)
A
(c)
Q
ZZ-ZZZZ
A/i ii i) i irn
///r/zllllllll III / / / /
Includes bacteria, algae, protozoa, and
other microscopic animals, and often the
young or embryonic stages of algae and
other organisms that normally grow up
to become a part of the benthos (see below).
Many planktonic types will also adhere
to surfaces as periphyton, and some
typical periphyton may break off and
be collected as plankters.
C Benthos are the plants and animals living
on, in, or closely associated with the
bottom. They include plants and
invertebrates.
D Nekton are the community of strong
aggressive swimmers of the open waters,
often called pellagic. Certain fishes,
whales, and invertebrates such as
shrimps and squids are included here.
E The marsh community is based on
larger "higher" plants, floating and
emergent. Both marine and freshwater
marshes are areas of enormous bio-
logical production.
VI PRODUCTIVITY
V BIOTIC COMMUNITIES
A Plankton are the macroscopic and
microscopic animals, plants, bacteria,
etc., floating free in the open water.
Many clog filters, cause tastes, odors,
and other troubles in water supplies.
Eggs and larvae of larger forms are
often present.
1 Phytoplankton are plant-like. These
are the dominant producers of the
waters, fresh and salt, "the grass
of the seas".
2 Zooplankton are animal-like.
Includes many different animal types,
range in size from minute protozoa
to gigantic marine jellyfishes.
B Periphyton (or Aufwuchs) - The communities
of microscopic organisms associated with
submerged surfaces of any type or depth.
A The biological resultant of all physical
and chemical factors is the quantity of
life that may actually be present. The
ability to produce this "biomass" is
often referred to as the "productivity"
of a body of water. This is neither good
nor bad per se. A water of low pro-
ductivity is a "poor" water biologically,
and also a relatively "pure" or "clean"
water, hence desirable as a water supply
or a bathing beach. A productive water
on the other hand may be a nuisance to
man or highly desirable. It is a nuisance
if foul odors and/or weed-chocked
waterways result, it ls desirable if
bumper crops of bass, catfish, or
oysters are produced. Open oceans on
the other hand have a very low level of
productivity in general
-------
The Aquatic Environment
REFERENCES
1 Clarke, G. L. Elements of Ecology.
John Wiley & Sons, New York. 1954.
2 Cooke, W. B. Trickling Filter Ecology.
Ecology 40(2):273-291. 1959.
3 Hanson, E. D. Animal Diversity.
Prentice-Hall, Inc., New Jersey. 1964.
4 Hedgpeth, J.W. Aspects of the Estuarine
Ecosystem. Amer. Fish. Soc., Spec.
Publ. No. 3. 1966.
5 Odum, E.P. Fundamentals of Ecology.
W.B. Saunders Company,
Philadelphia and London. 1959.
6 Patten, B.C. Systems Ecology
Bio-Science. 16(9). 1966.
7 Whittaker, R.H. New Concepts of
Kingdoms. Science 163:150-160. 1969.
This outline was prepared by H. W. Jackson,
Chief Biologist, National Training Center,
Water Programs Operations, EPA,
Cincinnati, OH 45268.
-------
THE AQUATIC ENVIRONMENT
Part 3. The Freshwater Environment
I INTRODUCTION
The freshwater environment as considered
herein refers to those inland waters not
detectably diluted by ocean waters, although
the lower portions of rivers are subject to
certain tidal flow effects.
Certain atypical inland waters such as saline
or alkaline lakes, springs, etc., are not
treated, as the main objective is typical
inland water.
All waters have certain basic biological cycles
and types of interactions most of which have
already been presented. Hence this outline
will concentrate on aspects essentially
peculiar to fresh inland waters.
II PRESENT WATER QUA LITY AS A
FUNCTION OF THE EVOLUTION OF
FRESH WATERS
A The history of a body of water determines
its present condition. Natural waters have
evolved in the course of geologic time
into what we know today.
B Streams
In the course of their evolution, streams
in general pass through four general
stages of development which may be
called, birth, youth, maturity, and old
age.
1 Establishment or birth. In an extant
stream, this might be a "dry run" or
headwater stream-bed, before it had
eroded down to the level of ground
water.
2 Youthful streams, when the stream-
bed is eroded below the ground water
level, spring water enters and the
stream becomes permanent.
3 Mature streams, have wide valleys,
a developed flood plain, deeper,
more turbid, and usually warmer
water, sand, mud, silt, or clay
bottom materials which shift with
increase in flow.
4 In old age, streams have approached
geologic base level. During flood
stage they scour their beds and deposit
materials on the flood plain which
may be very broad and flat. During
normal flow the channel is refilled
and many shifting bars are developed.
(Under the influence of man this
pattern may be broken up, or
temporarily interrupted. Thus an
essentially "youthful" stream might
take on some of the characteristics
of a "mature" stream following soil
erosion, organic enrichment, and
increased surface runoff. Correction
of these conditions might likewise be
followed by at least a partial reversion
to the "original" condition).
C Lakes and Reservoirs
Geological factors which significantly
affect the nature of either a stream or
lake include the following
1 The geographical location of the
drainage basin or watershed.
2 The size and shape of the drainage
basin.
3 The general topography, i.e.,
mountainous or plains.
4 The character of the bedrocks and
soils.
5 The character, amount, annual
distribution, and rate of precipitation.
BI. 21d. 9. 71
-------
The Aquatic Environment
The natural vegetative cover of the
land is of course responsible to many
of the above factors and is also
severely subject to the whims of
civilization. This is one of the major
factors determining runoff versus
soil absorption, etc.
D Lakes have a developmental history which
somewhat parallels that of streams.
1 The method of formation greatly
influences the character and sub-
sequent history of lakes.
2 Maturing or natural eutrophication of
lakes.
a If not already present shoal areas
are developed through erosion of
the shore by wave action and
undertow.
b Currents produce bars across bays
and thus cut off irregular areas.
c Silt brought in by tributary streams
settles out in the quiet lake water.
d Rooted aquatic plants grow on
shoals and bars, and in doing so
cut off bays and contribute to the
filling of the lake.
e Dissolved carbonates and other
materials are precipitated in the
deeper portions of the lake in part
through the action of plants.
f When filling is well advanced,
mats of sphagnum moss may extend
outward from the shore. These
mats are followed by sedges and
grasses which finally convert the
lake into a marsh.
3 Extinction of lakes. After lakes reach
maturity, their progress toward
filling up is accelerated. They become
extinct through
a The downcutting of the outlet.
Filling with detritus eroded from
the shores or brought in by
tributary streams.
Filling by the accumulation of the
remains of vegetable materials
growing in the lake itself.
(Often two or three processes may
act concurrently)
III PRODUCTIVITY IN FRESH WATERS
A Fresh waters in general and under
natural conditions by definition have a
lesser supply of dissolved substances
than marine waters, and thus a lesser
basic potential for the growth of aquatic
organisms. By the same token, they
may be said to be more sensitive to the
addition of extraneous materials
(pollutants, nutrients, etc.) The
following notes are directed toward
natural geological and other environ-
mental factors as they affect the
productivity of fresh waters.
B Factors Affecting Stream Productivity
(See Table 1)
TABLE 1
EFFECT OF SUBSTRATE ON STREAM
PRODUCTIVITY*
(The productivity of sand bottoms is
taken as 1)
Bottom MaienaL
Relative Productivity
Sand
1
Mail
6
Fine Gravel
q
Gravel and silt
14
Coarse gravel
32
Moss on fine gravel
B9
Fissidens (moss) on coarse gravel
11 1
Ranunculus (water buttercup)
194
Watercress
301
Anacharis (waterweed)
45 2
-'Selected from Tarzwell 1937
To be productive of aquatic life, a
stream must provide adequate nutrients,
light, a suitable temperature, and time
for growth to take place.
-------
The Aquatic Environment
1 Youthful streams, especially on rock
or sand substrates are low in essential
nutrients. Temperatures in moun-
tainous regions are usually low, and
due to the steep gradient, time for
growth is short. Although ample
light is available, growth of true
plankton is thus greatly limited.
2 As the stream flows toward a more
"mature" condition, nutrients tend to
accumulate, and gradient diminishes
and so time of flow increases, tem-
perature tends to increase, and
plankton flourish.
Should a heavy load of inert silt
develop on the other hand, the
turbidity would reduce the light
penetration and consequently the
general plankton production would
diminish.
3 As the stream approaches base level
(old age) and the time available for
plankton growth increases, the
balance between turbidity, nutrient
levels, and temperature and other
seasonal conditions, determines the
overall productivity.
C Factors Affecting the Productivity of
Lakes
1 The size, shape, and depth of the lake
basin. Shallow water is more pro-
ductive than deeper water since more
light will reach the bottom to stimulate
rooted plant growth. Asa corollary,
lakes with more shoreline, having
more shallow water, are in general
more productive. Broad shallow lakes
and reservoirs have the greatest
production potential (and hence should
be avoided for water supplies).
2 Hard waters are generally more
productive than soft waters as there
are more plant nutrient minerals
available. This is often greatly in-
fluenced by the character of the soil
and rocks in the watershed and the
quality and quantity of ground water
entering the lake. In general, pH
ranges of 6. 8 to 8. 2 appear to be
most productive.
TABLE 2
EFFECT OF SUBSTRATE
ON LAKE PRODUCTIVITY *
(The productivity of sand bottoms is taken as 1)
Bottom Material
Relative Productivity
Sand
1
Pebbles
4
Clay
8
Flat rubble
9
Block rubble
11
Shelving rock
77
^Selected from Tarzwell 1937
3 Turbidity reduces productivity as
light penetration is reduced.
4 The presence or absence of thermal
stratification with its semi-annual
turnovers affects productivity by
distributing nutrients throughout the
water mass.
5 Climate, temperature, prevalence of
ice and snow, are also of course
important.
D Factors Affecting the Productivity of
Reservoirs
1 The productivity of reservoirs is
governed by much the same principles
as that of lakes, with the difference
that the water level is much more
under the control of man Fluctuations
in water level can be used to de-
liberately increase or decrease
productivity. This can be
demonstrated by a comparison of
the TVA reservoirs which practice
a summer drawdown with some of
those in the west where a winter
drawdown is the rule.
2 The level at which water is removed
from a reservoir is important to the
productivity of the stream below
-------
The Aquatic Environment
The hypolimnion may be anaerobic
while the epilimnion is aerobic, for
example, or the epilimnion is poor in
nutrients while the hypolimnion is
relatively rich.
3 Reservoir discharges also profoundly
affect the DO, temperature, and
turbidity in the stream below a dam.
Too much fluctuation irj flow may
permit sections of the stream to dry,
or provide inadequate dilution for
toxic waste.
VII CLASSIFICATION OF LAKES AND
RESERVOIRS
A The productivity of lakes and impound-
ments is such a conspicuous feature that
it is often used as a convenient means of
classification.
1 Oligotrophic lakes are the younger,
less productive lakes, which are deep,
have clear water, and usually support
Salmonoid fishes in their deeper waters.
2 Eutrophic lakes are more mature,
more turbid, and richer They are
usually shallower They are richer
in dissolved solids, N, P, and Ca are
abundant. Plankton is abundant and
there is often a rich bottom fauna.
3 Dystrophic lakes, such as bog lakes,
are low in pH, water yellow to brown,
dissolved solids, N, P, and Ca scanty
but humic materials abundant, bottom
fauna and plankton poor, and fish
species are limited.
B Reservoirs may also be classified as
storage and run of the river.
1 Storage reservoirs have a large
volume in relation to their inflow.
2 Run of the river reservoirs have a
large flow-through in relation to their
storage value.
C According to location, lakes and
reservoirs may be classified as polar,
temperate, or tropical. Differences in
climatic and geographic conditions
result in differences in their biology.
VIII SUMMARY
A A body of water such as a lake, stream,
or estuary represents an intricately
balanced system in a state of dynamic
equilibrium. Modification imposed at
one point in the system automatically
results in compensatory adjustments at
associated points.
B The more thorough our knowledge of the
entire system, the better we can judge
where to impose control measures to
achieve a desired result.
REFERENCES
1 Chamberlin, Thomas C. and Salisburg,
Rollin P Geological Processes and
Their Results. Geology 1 pp i-xdc,
and 1-654. Henry Holt and Company.
New York. 1904.
2 Frey, David G. Limnology in North
America. Univ. Wise. Press. 1963.
3 Hutcheson, George E. A Treatise on
Limnology Vol. I Geography, Physics
and Chemistry, 1957. Vol. II.
Introduction to Lake Biology and the
Limnoplankton. 1115 pp. 1967.
John Wiley Co.
4 Hynes, H.B.N. The Ecology of Running
Waters. Univ. Toronto Press.
555 pp. 1970.
5 Ruttner, Franz. Fundamentals of
Limnology. University of Toronto
Press, pp. 1-242. 1953.
-------
The Aquatic Environment
6 Tarzwell, Clarence M. Experimental
Evidence on the Value of Trout 1937
Stream Improvement in Michigan.
American Fisheries Society Trans.
66:177-187. 1936.
8 Ward and Whipple. Fresh Water
Biology. (Introduction). John
Wiley Company. 1918.
7 U.S. Dept. of Health, Education, and
Welfare. Public Health Service.
Algae and Metropolitan Wastes.
Transactions of a seminar held
April 27-29, I960 at the Robert A.
Taft Samtary Engineering Center.
Cincinnati, OH. No. SEC TR W61-3.
This outline was prepared by H.W. Jackson,
Chief Biologist, National Training Center,
Water Programs Operations, EPA, Cincinnati,
OH 452 68.
-------
THE AQUATIC ENVIRONMENT
Part 4. The Marine Environment and its Role in the Total Aquatic Environment
I INTRODUCTION
A The marine environment is arbitrarily
defined as the water mass extending
beyond the continental land masses,
including the plants and animals harbored
within. This water mass is large and
deep, covering about 70 percent of the
earth's surface and being as deep as
7 miles. The salt content averages
about 35 parts per thousand. Life extends
to all depths.
B The general nature of the water cycle on
earth is well known. Because the rel-
atively large surface area of the earth
is covered with water, roughly 70 percent
of the earth's rainfall is on the seas.
(Figure 1)
70*
rUun 1. THR VATK8 CIC1I
Since roughly one third of the earth's
rain which falls on the land is again
recycled through the stratosphere
(see Figure 1 again), the total amount
of water washing over the earth's surface
is significantly greater than one third of
the total world rainfall. It is thus not
surprising to note that the rivers which
finally empty into the sea carry a con-
siderable burden of dissolved and
suspended solids picked up from the land.
This is the substance of geological
erosion. (Table 1)
TABLE 1
PERCENTAGE COMPOSITION OF THE MAJOR IONS
OF TWO STREAMS AND SEA WATER
(Data from Clark, F.W., 1924, "The Composition of River
and Lake Waters of the United States", U.S. Geol. Surv.,
Prof. Paper No. 135, Harvey, H.W., 1957, "The Chemistry
and Fertility of Sea Waters", Cambridge University Press,
Cambridge)
Ion
Delaware River
at
Lambertville, N.J.
Rio Grande
at
Laredo, Texas
Sea Water
Na
6 70
14.78
30 4
K
1.46
85
1. 1
Ca
17.49
13.73
1. 16
Mg
4 81
3.03
3 7
CI
4 23
21 65
55 2
so4
17 49
30. 10
7 7
C°3
32. 95
11.55
mco3 0 35
C For this presentation, the marine
environment will be (1) described using
an ecological approach, (2) characterized
ecologically by comparing it with fresh-
water and estuarine environments, and
(3) considered as a functional ecological
system (ecosystem).
II FRESHWATER, ESTUARINE. AND
MARINE ENVIRONMENTS
Distinct differences are found in physical,
chemical, and biotic factors in going from
a freshwater to an oceanic environment.
In general, environmental factors are more
constant in freshwater (river) and oceanic
environments when compared to the highly
variable and harsh environments of estuarine
and coastal waters.
A Physical and Chemical Factors
(Figure 2)
1 Rivers
2 Estuary and coastal waters
3 Oceans
-------
The Aquatic Environment
Degree of instability
Avail-
ability
of
nutrients
(degree)
Type of environment
and general direction
of water movement
Salinity
Temperature
Water
elevation
Vertical
strati-
fication
Turbidity
?!'
1
1
-
-
BB
—
( \+
/Eatuarine \
3 J
—
—
-
—
—
-
Oceanic 4^"r
¦
¦
¦
¦
¦
I
Figure2 . RELATIVE VALUES OF VARIOUS PHYSICAL AND CHEMICAL FACTORS
FOR RIVER, ESTUARINE, AND OCEANIC ENVIRONMENTS
B Biotic Factors
1 A complex of physical and chemical
factors determine the biotic composi-
tion of an environment. In general,
the number of species in a highly
variable environment tends to be less
than the number in a more stable
environment (Hedgpeth, 1966).
2 The dominant animal species (in
terms of total biomass) which occur
in estuaries are often transient,
spending only a part of their lives in
the estuaries. This results in better
utilization of a rich environment.
C Zones of the Sea
The nearshore environment is often
classified in relation to tide level and
water depth. The nearshore and oceanic
regions together are often classified in
relation to light penetration and water
depth.
1 Neritic - Relatively shallow-water
zone which extends from the high-
tide mark to the edge of the
continental shelf. (Figure 3)
-------
The Aquatic Environment
Ben thai
Pelagia)
__ Neritic
Region
Illuminated —
Dark _
200
400
-J 600
Primaiy subdivisions of the marine habitat.
Figure 3.
a Stability of physical factors is
intermediate between estuarine
and oceanic environments.
b Phytoplankters are the dominant
producers but in some locations
attached algae are also important
as producers.
c The animal consumers are
zooplankton, nekton, and benthic
forms.
Oceanic - The region of the ocean
beyond the continental shelf. Divided
into three parts, all relatively
poorly populated compared to the
neritic zone.
1) Physical factors fluctuate
less than in the neritic zone.
2) Producers are the phyto-
plankton and consumers are
the zooplankton and nekton.
b Bathyal zone - From the bottom
of the euphotic zone to about
6, 000 feet.
1) Physical factors relatively
constant but light is absent.
2) Producers are absent and
consumers are scarce.
c Abyssal zone - All the sea below
the bathyal zone.
Euphotic zone - Waters into which
sunlight penetrates (often to the
bottom m the neritic zone). The
zone of basic productivity. Often
extends to 600 feet below the
surface.
1) Physical factors more con-
stant than in bathyal zone.
2) Producers absent and
consumers not as abundant
as in the bathyal zone.
-------
The Aquatic Environment
HI SEA WATER AND THE BODY FLUIDS
A Sea water is a most suitable environment
for living cells, because it contains all
of the chemical elements essential to the
growth and maintenance of plants and
animals. The ratio and often the con-
centration of the major salts of sea water
are strikingly similar in the cytoplasma
and body fluids of marine organisms.
This similarity is also evident, although
modified somewhat in the body fluids of
both fresh water and terrestrial animals.
For example, sea water may be used in
emergencies as a substitute for blood
plasma in man.
B Since marine organisms have an internal
salt content similar to that of their
surrounding medium (isotonic condition)
osmoregulation poses no problem. On the
other hand, fresh water organisms are
hypertonic (osmotic pressure of body
fluids is higher than that of the surround-
ing water). Hence, fresh water animals
must constantly expend more energy to
keep water out (i. e., high osmotic
pressure fluids contain more salts, the
action being then to dilute this concen-
tration with more water).
1 Generally, marine invertebrates are
narrowly poikilosmotic, i.e., the salt
concentration of the body fluids changes
with that of the external medium. This
has special significance in estuarine
situations where salt concentrations
of the water often vary considerably
in short periods of time.
2 Marine bony fish (teleosts) have lower
salt content internally than externally
(hypotonic). In order to prevent
dehydration, water is ingested and salts
are excreted through special cells in
the gills.
IV
FACTORS AFFECTING THE DISTRI-
BUTION OF MARINE ORGANISMS
Salinity - The concentration of salts is
not the same everywhere in the sea; in
the open ocean salinity is much less
variable than in the ever changing
estuary or coastal water. Organisms
have different tolerances to salinity
which limit their distribution. The
distributions may be in large water
masses, such as the Gulf Stream,
Sargasso Sea, etc., or in bays and
estuaries.
1 In general, animals m the estuarine
environment are able to withstand
large and rapid changes in salinity
and temperature. These animals are
classified as
a Euryhaline ("eury" meaning wide)
wide tolerance to salinity changes.
EURYHALINE
Freeh Water
Stenohaline
Marine
Stenohaline
Salinity
ca. 35
Figure 4. Salinity Tolerance of Organisms
b Eurythermal - wide tolerance to
temperature changes.
-------
The Aquatic Environment
a
0
0
6
o
©
ffl
Figure 5
Zonation of plants, snails, and barnacles on a rocky shore. While
this diagram is based on the situation on the southwest coast of
England, the general idea of zonation may be applied to any temper-
ate rocky ocean shore, though the species will differ. The gray
zone consists largely of lichens. At the left is the zonation of rocks
with exposure too extreme to support algae; at the right, on a less
exposed situation, the animals are mostly obscured by the algae.
Figures at the right hand margin refer to the percent of time that
the zone is exposed to the air, i. e., the time that the tide is out.
Three major zones can be recognized: the Littorina zone (above the
gray zone); the Balanoid zone (between the gray zone and the
laminarias); and the Laminaria zone. a. Pelvetia canaliculata;
b. Fucus spiralis; c. Ascophyllum nodosum; d. Fucus serratus;
e. Laminaria digitata. (Based on Stephenson)
SNAILS
Littorina neritoides
L. rudis
L. obtusata
L. littorea
BAiiNACLES
Chthamalus stellatus
Balanus balanoides
B. perforatus
-------
The Aquatic Environment
2 In general animals in river and
oceanic environments cannot withstand
large and rapid changes in salinity and
temperature. These animals are
classified as
a Stenohaline {"steno" meaning narrow) -
narrow tolerance to salinity changes.
b Stenothernal - narrow tolerance to
temperature changes.
3 Among euryhaline animals, those living
in lowered salinities often have a
smaller maximum size than those of
the same species living in more saline
waters. For example, the lamprey
(Petromyzon marinus) attains a length
of 30 - 36 in the sea, while in the
Great Lakes the length is 18 - 24".
4 Usually the larvae of marine organisms
are more sensitive to changes in salinity
than are the adults This character-
istic limits both the distribution and
size of populations.
B Tides
Tidal fluctuation is a phenomenon unique
to the seas (with minor exceptions). It is
a twice daily rise and fall m the sea level
caused by the complicated interaction of
many factors including sun, moon, and the
daily rotation of the earth. Tidal heights
vary from day to day and place to place,
and are often accentuated by local
meteorological conditions. The rise and
fall may range from a few inches or less
to fifty feet or more.
V FACTORS AFFECTING THE
PRODUCTIVITY OF THE MARINE
ENVIRONMENT
The sea is in continuous circulation. With-
out circulation, nutrients of the ocean would
eventually become a part of the bottom and
biomass production would cease. Generally,
in all oceans there exists a warm surface
layer which overlies the colder water and
forms a two-layer system of persistent
stability. Nutrient concentration is usually
greatest in the lower zone. Wherever a
mixing or disturbance of these two layers
occurs, biomass production is greatest.
Factors causing this breakup are, therefore,
of utmost importance concerning productivity.
ACKNOWLEDGEMENT-
This outline contains selected material
from other outlines prepared by C.M.
Tarzwell, Charles L. Brown, Jr.,
C.G. Gunnerson, W.Lee Trent, W.B.
Cooke, B. H. Ketchum, J.K. McNulty,
J. L. Taylor, R. M. Sinclair, and others.
REFERENCES
1 Harvey, H. W. The Chemistry and
Fertility of Sea Water (2nd Ed.).
Cambridge Univ. Press, New York.
234 pp. 1957.
2 Hedgpeth, J.W. (Ed.). Treatise on
Marine Ecology and Paleoecology.
Vol. I. Ecology Mem. 67 Geol.
Soc. Amer., New York. 1296 pp.
1957.
3 Hill, M.N. (Ed.). The Sea. Vol. II.
The Composition of Sea Water
Comparative and Descriptive
Oceanography. Interscience Pubis.
John Wiley & Sons, New York.
554 pp. 1963.
4 Moore, H. B. Marine Ecology. John
Wiley &. Sons, Inc., New York.
493 pp. 1958.
5 Reid, G.K. Ecology of Inland Waters
and Estuaries. Reinhold Publ.
Corp. New York. 375 pp. 1961.
6 Sverdrup, Johnson, and Fleming.
The Oceans. Prentice-Hall, Inc.,
New York. 1087 pp. 1942.
This outline was prepared by H.W. Jackson,
Chief Biologist, National Training Center,
Water Programs Operations, EPA, Cincinnati,
OH 45268.
-------
THE AQUATIC ENVIRONMENT
Part 5: Tidal Marshes
I INTRODUCTION: The Marsh and the
Estuary
A "There is no other case in nature, save
in the coral reefs, where the adjustment
of organic relations to physical condition
is seen in such a beautiful way as the
balance between the growing marshes
and the tidal streams by which they are
at once nourished and worn away. "
(Shaler, 1886)
B Estuarine pollution studies are usually
devoted to the dynamics of the circulating
water, its chemical, physical, and
biological parameters, bottom deposits, etc.
C It is easy to overlook the intimate relation-
whips which exist between the bordering
marshland, the moving waters, the tidal
flats, subtidal deposition, and seston
whether of local, oceanic, or riverine
origin.
D The tidal marsh (some inland areas also
have salt marshes) is generally considered
to be the marginal areas of estuaries and
coasts in the intertidal zone which are
dominated by emergent vegetation. They
generally extend inland to the farthest
point reached by the spring tides, where
they merge into freshwater swamps and
marshes (Figure 1). They may range in
width from nonexistent on rocky coasts to'
many kilometers.
H MARSH ORIGINS AND STRUCTURES
A In general, marsh substrates are high in
organic content, relatively low in minerals
and trace elements. The upper layers
bound together with living roots called
turf, underlaid by more compacted peat
type material.
1 Rising or eroding coastlines may
expose peat from ancient marsh
growth to wave action which cuts
into the soft peat rapidly (Figure 2).
Such banks are likely to be cliff-like,
and are often undercut. Chunks of
peat are often found lying about on
harder substrate below high tide line.
If face of cliff is well above high water,
overlying vegetation is likely to be
typically terrestrial of the area.
Marsh type vegetation is probably
absent.
2 Low lying deltaic, or sinking coast-
lines, or those with low energy wave
action are likely to have active marsh
formation in progress (Figure 3).
Sand dunes are also common in such
areas (Figure 4). General coastal
configuration is a factor.
a Rugged or precipitous coasts or
slowly rising coasts, typically
exhibit narrow shelves, sea cliffs,
fjords, massive beaches, and
relatively less marsh area (Figure 5).
An Alaskan fjord subject to recent
catastrophic subsidence and rapid
deposition of glacial flour shows
evidence of the recent encroachment
of saline waters in the presence of
recently buried trees and other
terrestrial vegetation, exposure
of layers of salt marsh peat along
the edges of channels, and a poorly
compacted young marsh turf developing
at the new high water level (Figure 6).
b Low lying coastal plains tend to be
fringed by barrier islands, broad
estuaries and deltas, and broad
associated marshlands (Figure 7, 14).
Deep tidal channels fan out through
innumerable branching and often
interconnecting rivulets. The
intervening grassy plains are
essentially at mean high tide level.
BI. 21d. 9.71
-------
CO
o
Freshwater
Mud Flat
Tidal Marsh
^ Marsh ^
Spartina
Juncus
Trans ltional
Blue Clay
Figure 1 Zonation in a positive New England estuary 1. Spring tide level, 2. Mean high tide,
3. Mean low tide, 4. Bog hole, 5. Ice cleavage pool, 6. Chunk of Spartina turf deposited by ice,
7. Organic ooze with associated community, 8. eelgrass (Zostera), 9. Ribbed mussels (modiolus)-
-------
The Aquatic Environment
/i! MVf)\%
Figure 2. Diagrammatic section of eroding peat cliff
>E
UJtt Y' ^i-,J'. -^ aSsan wrjojia.C' fr.lTr~ Yr!":?'
Figure 3. Effects of deltaic subsidence
during distributary system abandonment
c Tropical and subtropical regions
such as Florida, the Gulf Coast,
and Central America, are frequented
by mangrove swamps. This unique
type of growth is able to establish
itself in shallow water and move out
into progressively deeper areas
(Figure 8). The strong deeply
embedded roots enable the mangrove
to resist considerable wave action
at times, and the tangle of roots
quickly accumulates a deep layer of
organic sediment. Mangroves are
often considered to be effective as
land builders. When fully developed,
a mangrove swamp is an impene-
trable thicket of roots over the tidal
flat affording shelter to a sort of
semi-aquatic organism such as
various molluscs and crustaceans,
and providing access from the
nearby land to predaceous birds,
reptiles and mammals.
Mangroves are not restricted to
estuaries, but may develop out into
shallow oceanic lagoons, or upstream
into relatively fresh waters.
in PRODUCTIVITY OF MARSHES
A Measuring the productivity of grasslands
is not easy, because grass is seldom used
directly as such by man. It is thus
usually expressed as production of meat,
milk, or in the case of salt marshes, the
total crop of animals that obtain food per
unit of area. The primary producer in a
tidal marsh is the marsh grass, but very
little of it is used as grass. (Table 1)
The actual nutritional analysis of several
marsh grasses as compared to dry land
bay is shown in Table 2. A study of the
yield of Juncus per square meter in a
North Carolina marsh is shown in Figure 9.
B The actual utilization of marsh grass is
accomplished primarily by its decom-
position and ingestion by micro flora and
fauna. A small quantity of seeds and
solids is probably consumed directly by
birds (Figure 10).
1 The quantity of micro invertebrates which
thrive on this wealth of decaying marsh
hay has not been estimated, nor has the
actual production of small fishes such
as tb' ;_,p minnows (Fundulus) which
swarm in at high tide, or the mud
snails (Nassa) and others. Many forms
of oceanic life migrate into the estuaries,
especially the marsh areas, for impor-
tant portions of their life histories as
has been mentioned elsewhere (Figure 11).
-------
The Aquatic Environment
MHW -18 1300SBC
Figure 4
Development of a Massachusetts Marsh since 1300 BC, involving an
18 foot rise in water level. Shaded area indicates sand dunes. Note
meandering marsh tidal drainage. A: 1300 BC, B: 1950 AD.
-------
The Aquatic Environment
NEWPORT
OREGON
Figure 5. A River Mouth on a Slowly Rising Coast. Note absence
of deltaic development and relatively little marshland,
although mud flats stippled are extensive.
2 An indirect approach in Rhode Island
revealed in a single August day on a
relatively small marsh area, between
700 and 1000 wild birds of 12 species,
ranging from 100 least sandpipers to
uncountable numbers of seagulls. One
food requirement estimate for three-
pound poultry in the confined inactivity
of a poultry yard is approximately one
ounce per pound of bird per day.
One-hundred (100) black bellied plovers
at approximately ten (10) ounces each
would weigh on the order of sixty (60)
pounds. At the same rate of food
consumption, this would indicate nearly
four (4) pounds of food required for
this species alone. The much
greater activity of the wild birds
would obviously greatly increase their
food requirements, as would their
relatively smaller size.
Considering the range of foods con-
sumed, the sizes of the birds, and the
fact that at certain seasons, thousands
of migrating ducks and others pause
to feed here, the enormous productivity
of such a marsh can be better under-
stood.
-------
The Aquatic Environment
Terrestria I
Tidal marsh
Shifting flats
Figure 6. Some general relationships in a northern fjord with a rising water level. 1. mean low
water, 2. maximum high tide, 3. Bedrock, 4. Glacial flour to depths in excess of
400 meters, 5. Shifting flats and channels, 6. Channel against bedrock, 7. Buried
terrestrial vegetation, 8. Outcroppings of salt marsh peat.
Figure 7. A Coastal Plain Marsh
in India subject to a high
tidal range.
r 1If mil.
TROPICAL CONOCARPUS AVICENNIA RHIZOPHORA
FOREST TRANSITION ASSOCIES SALT-MARSH ASSOCIES CONSOOES
Figure 8. Diagrammatic transect of a mangrove swamp
showing transition from marine to terrestrial
habitat.
-------
The Aquatic Environment
TABLE 1. General Orders of Magnitude of Gross Primary Productivity in Terms
of Dry Weight of Organic Matter Fixed Annually
Ecosystem
gms/M /year
(grams/square meters/year)
lbs/acre/year
Land deserts, deep oceans Tens
Grasslands, forests, cutrophic Hundreds
lakes, ordinary agriculture
Estuaries, deltas, coral reefs. Thousands
intensive agriculture (sugar
cane, rice)
Hundreds
Thousands
Ten-thousands
TABLE 2. Analyses of Some Tidal Marsh Grasses
T/A Petccntane Composition
Dry Wt Proiein fat fiber Water
Ash
N-free Extract
DisnrMis spicarj (pure stand, dry)
28 53 17 32 4 82 6 7
Short Spartma jllcrniilorj and Sohcorilio t'uropoej (in standinf; water)
1 2 7 7 2 5 31 1 8 8 12 0
Spjrhns jltcrnillrjrj (tall, pure stand in standing water)
3 5 7 f, 2 0 29 0 8 3 15 5
Spjr'ind pd" ns 'u'l'' s'.ind, dry)
J 2 1,0 11 JO 0 81 9 0
Spirimj a/i'vnif/oM and Sp.iriitu (mined stand, wet)
i 4 61! 1 'J 2*J 1) (11
Sp.iHin.) .ihrrniWiu (short, wet)
11 0 fl 2 4
Comparable Analyses for Hay
1 t i ui r. 0 2 0
Aid nit 110 17
20 1)
30 4
V, 2
211 5
I) 7
fi 7
10 4
104
13 3
4 2
ri<)
45 5
37 7
17 3
44 5
428
36 3
44 9
«) S
Analyses performed by Roland W, Gilbert, Department
of Agricultural Chemistry, U. R. I
-------
The Aquatic Environment
2.200
1 800
1 400
o.
O
cc
u
a
z
o
z
1 000
600
Dying
400
Live
JULY
Figure 9. Standing crop of Juncus. Solid line represents observed
values, broken line represents seasonal cycle calculated
on the basis of an assumed constant total biomass.
SPARTlNJk
!MARSH|
N-FRCt (XTRAC1
YOUNG
CRUDE FIBfiC
[SOUNDl
lAflVAl
/ STAGES
IQPENr OCEAN]
EGGS
ADULT
Figure 10. The nutritive composition of
successive stages of decomposition of
Spartina marsh grass, showing increase
in protein and decrease in carbohydrate
with increasing age and decreasing size
of detritus particles.
Figure 11. Diagram of the life cycle
of white shrimp (after Anderson and
Lunz 1965).
-------
The Aquatic Environment
Greater yellow legs (left)
and black duck
Great blue heron
Figure 12. Some Common Marsh Birds
5 Odurn, E.P. The Role of Tidal Marshes
in Marine Production. The Conservationist
(NY), June-July. 1961.
6 Odum, E.P. and Dela Crug, A.A.
Particulate Organic Detritus m a
Georgia Salt Marsh - Estuarine
Ecosystem, in: Estuaries, pp. 383-
388, Publ. No. 83, Am. Assoc. Adv.
Sci. Washington, DC. 1967.
7 Redfield, A. C. The Ontogeny of a Salt
Marsh Estuary.^n: Estuaries, pp.
108-114. Publ. No. 83, Am. Assoc.
Adv. Sci. Washington, DC. 1967,
8 Stuckey, O.H. Measuring the Productivity
of Salt Marshes. Mantimes (Grad
School of Ocean., U.R.I.) Vol. 14(1)-
9-11. February 1970.
REFERENCES 9
1 Anderson, W.W. The Shrimp and the
Shrimp Fishery of the Southern
United States. USDI, FWS, BCF.
Fishery Leaflet 589. 1966.
2 Dewey, E. S., Jr. Bogs. Sci. Am. Vol.
100 (4): 115-122. October 1958.
Williams, R.B. Compartmental
Analysis of Production and Decay
of Juncus roemerianus. Prog.
Report, Radiobiol. Lab., Beaufort, NC,
Fiscal Year 1968, USDI, BCF, pp. 10-
12.
3 Emery, K, O. and Stevenson. Estuaries
and Lagoons. Part n, Biological
Aspects by J. W. Hedgepeth, pp. 693-
728. in: Treatise on Marine Ecology
and Paleoecology. Geol. Soc. Am.
Mem. 67. Washington, DC. 1957.
4 Morgan, J. P. Ephemeral Estuaries of the
Deltaic Environment irr Estuaries,
pp. 115-120. Publ. No. 83, Am.
Assoc. Adv. Sci. Washington, DC. 1967.
This outline was prepared by H.W. Jackson,
Chief Biologist, National Training Center,
Water Programs Operations, EPA, Cincinnati,
OH 45268.
-------
SIGNIFICANCE OF "LIMITING FACTORS" TO POPULATION VARIATION
I INTRODUCTION
A All aquatic organisms do not react uniformly
to the various chemical, physical and
biological features in their environment
Through normal evolutionary processes
various organisms have become adapted
to certain combinations of environmental
conditions. The successful development
and maintenance of a population or community
depend upon harmonious ecological balance
between environmental conditions and
tolerance of the organisms to variations
in one or more of these conditions.
B A factor whose presence or absence exerts
some restraining influence upon a population
through incompatibility with species
requirements or tolerance is said to be a
limiting factor. The principle of limiting
factors is one of the major aspects of the
environmental control of aquatic organisms
(Figure 1).
A Liebig's Law of the Minimum enunciates
the first basic concept In order for an
organism to inhabit a particular environ-
ment, specified levels of the materials
necessary for growth and development
(nutrients, respiratory gases, etc. ) must
be present. If one of these materials is
absent from the environment or present
in minimal quantities, a given species
will only survive in limited numbers, if
at all (Figure 2).
ui
u
z
¦<
a
z
3
CO
optimum
<
Ui
>
f-
<
UJ
ac
CRITICAL RANGE
LOW
MAGNITUDE OF FACTOR
HIGH
II PRINCIPLE OF LIMITING FACTORS
This principle rests essentially upon two basic
concepts. One of these relates organisms to
the environmental supply of materials essential
for their growth and development The second
pertains to the tolerance which organisms
exhibit toward environmental conditions.
Figure 2. Relationships of environmental
factors and the abundance of organisms.
1 The subsidiary principle of factor
interaction states that high concentration
or availability of some substance, or
the action of some factor in the environ-
ment, may modify utilization of the
minimum one. For example
/UNLIMITED GROWTH
/ DECREASE IN
iTWTfiTIOMNS
EQUILIBRIUM WITH
ENVIRONMENT
\
INCREASE IN
\ "TimTATions
* POPULATION DECLINE
TIME
The uptake of phosphorus by the
algae Nitzchia closterium is influenced
by the relative quantities of nitrate
and phosphate in the environment;
however, nitrate utilization appears
to be unaffected by the phosphate
(Reid, 1961).
The assimilation of some algae is
closely related to temperature.
Figure 1 The relationships of limiting factors
to population growth and development
The rate of oxygen utilization by fish
may be affected by many other sub-
stances or factors in the environment.
BI. ECO. 20a. 7. 69
-------
Signilicance of "Limiting Factors" to Population Variation
d Where strontium is abundant, mollusks
are able to substitute it, to a partial
extent, for calcium in their shells
(Odum, 1959).
2 If a material is present in large amounts,
but only a small amount is available for
use by the organism, the amount available
and not the total amount present deter-
mines whether or not the particular
material is limiting (calcium in the form
of CaC03).
B Shelford pointed out in his Law of Tolerance
that there are maximum as well as minimum
values of most environmental factors which
can be tolerated Absence or failure of an
organism can be controlled by the deficiency
or excess of any factor which may approach
the limits of tolerance for that organism
(Figure 3).
Figure 3. Shelford's Law of Tolerance.
1 Organisms have an ecological minimum
and maximum for each environmental
factor with a range in between called
the critical range which represents the
range of tolerance (Figure 2). The
actual range thru which an organism can
grow, develop and reproduce normally
is usually much smaller than its total
range of tolerance.
2 Purely deleterious factors (heavy metals,
pesticides, etc.) have a maximum
tolerable value, but no optimum (Figure 4).
HI
U
z
<
a
z
3
A
<
Ul
>
t-
<
-J
tu
at
CONCENTRATION
Figure 4. Relationship of purely harmful
factors and the abundance of
organisms.
3 Tolerance to environmental factors
varies widely among aquatic organisms.
a A species may exhibit a wide range
of tolerance toward one factor and a
narrow range toward another. Trout,
for instance, have a wide range of
tolerance for salinity and a narrow
range for temperature.
b All stages in the life history of an
organism do not necessarily have the
same ranges of tolerance. The
period of reproduction is a critical
time in the life cycle of most
organisms.
c The range of tolerance toward one
factor may be modified by another
factor. The toxicity of most sub-
stances increases as the temperature
increases.
d The range of tolerance toward a given
factor may vary geographically within
the same species. Organisms that
adjust to local conditions are called
ecotypes.
Minimum Limit of
Toleration
Range of Optimum
of Factors
Maximum Limit of
Toleration
Absent
Decreasing
Abundance
Greatest Abundance
Decreasing
Abundance
Absent
-------
Significance of "Limiting Factors" to Population Variation
e The range of tolerance toward a given
factor may vary seasonally. In general
organisms tend to be more sensitive
to environmental changes in summer
than in other seasons. This is
primarily due to the higher summer
temperatures.
4 A wide range of distribution of a species
is usually the result of a wide range of
tolerances. Organisms with a wide
range of tolerance for all factors are
likely to be the most widely distributed,
although their growth rate may vary
greatly. A one-year old carp, for
instance, may vary in size from less
than an ounce to more than a pound
depending on the habitat.
5 To express the relative degree of
tolerance for a particular environmental
factor the prefix eury (wide) or steno
(narrow) is added to a term for that
feature (Figure 5).
C The law of the minimum as it pertains to
factors affecting metabolism, and the law
of tolerance as it relates to density and
distribution, can be combined to form a
broad principle of limiting factors.
1 The abundance, distribution, activity
and growth of a population are deter-
mined by a combination of factors, any
one of which may through scarcity or
overabundance be limiting.
2 The artificial introduction of various
substances into the environment tends
to eliminate limiting minimums for
some species and create intolerable
maximums for others.
3 The biological productivity of any body
of water is the end result of interaction
of the organisms present with the
surrounding environment.
Ill VALUE AND USE OF THE PRINCIPLE OF
LIMITING FACTORS
STENOTHERMAL STENOTHERMAL
(OLIGOTHERMAL! CURT THERMAL (POLYTHERMAL)
TEMPERATURE
The organism-environment relationship
is apt to be so complex that not all factors
are of equal importance in a given situation;
some links of the chain guiding the organism
are weaker than others. Understanding
the broad principle of limiting factors and
the subsidiary principles involved make
the task of ferreting out the weak link in
a given situation much easier and possibly
less time consuming and expensive.
1 If an organism has a wide range of
tolerance for a factor which is
relatively constant in the environment
that factor is not likely to be limiting.
The factor cannot be completely
eliminated from consideration, however,
because of factor interaction.
Figure 5. Comparison of relative limits of
tolerance of stenothermal and
eurythermal organisms.
2 If an organism is known to have narrow
limits of tolerance for a factor which is
also variable in the environment, that
factor merits careful study since it
might be limiting.
-------
Significance of "Limiting Factors" to Population Variation
B Because of the complexity of the aquatic
environment, it is not always easy to
isolate the factor in the environment that
is limiting a particular population.
Premature conclusions may result from
limited observations of a particular
situations. Many important factors may
be overlooked unless a sufficiently long
period of time is covered to permit the
factors to fluctuate within their ranges of
possible variation. Much time and money
may be wasted on control measures jvithout
the real limiting factor ever being dis-
covered or the situation being improved.
C Knowledge of the principle of limiting
factors may be used to limit the number
of parameters that need to be measured or
observed for a particular study. Not all
of the numerous physical, chemical and
biological parameters need to be measured
or observed for each study undertaken.
The aims of a pollution survey are not to
make and observe long lists of possible
limiting factors but to discover which
factors are significant, how they bring
about their effects, the source or sources
of the problem, and what control measures
should be taken.
D Specific factors in the aquatic environment
determine rather precisely what kinds of
organisms will be present in a particular
area. Therefore, organisms present or
absent can be used to indicate environ-
mental conditions. The diversity of
organisms provides a better indication of
environmental conditions than does any
single species. Strong physio-chemical
limiting factors tend to reduce the diversity
within a community; more tolerant species
are then able to undergo population growth.
REFERENCES
1 Odum, Eugene P. Fundamentals of
Ecology, W. B. Saunders Company,
Philadelphia. (1959)
2 Reid, George K. Ecology of Inland Waters
and Estuaries. Reinhold Publishing
Corporation, New York. (1961)
This outline was prepared by John E.
Matthews, Aquatic Biologist, Robert S. Kerr
Water Research Center, Ada, Oklahoma.
-------
THE SYSTEM OF BIOLOGICAL CLASSIFICATION
I INTRODUCTION
There are few major groups of organisms
that are either exclusively terrestrial or
generally aquatic. The following remarks
apply to both, however, primary attention
will be directed to aquatic types.
II CLASSIFICATION
One of the first questions usually posed
about an organism seen for the first time
is "what is it'" usually meaning, "what is
its name' " The naming or classification of
biological organisms is a science in itself
(taxonomy). Some of the principles involved
need to be understood by anyone working
with organisms however.
A Names are the "key number", "code
designation", or "file references" which
we must have to find information about
an unknown organism.
B Why are they so long and why must they
be in Latin and Greek' File references
in large systems have to be long in order
to designate the many divisions and sub-
divisions. There are over a million and
a half items (or species) included in the
system of biological nomenclature (very
few libraries have as many as a million
books to classify).
C Common names are rarely available for
most invertebrates and algae. Exceptions
to this are common among the molluscs,
many of which have common names which
are fairly standard for the same species
throughout its range. This may be due
to their status as a commercial harvest
or to the activities of devoted groups of
amateur collectors. Certain scientific
societies have also assigned "official"
common names to particular species,
for example, aquatic weeds - American
Weed Society, fish - American Fisheries
Society, amphibians (salamanders and
frogs) - American Society of
Ichthyologists and Herpetologists.
D The system of biological nomenclature
is regulated by international congresses.
1 It is based on a system of groups and
super groups, of which the foundation
(which actually exists in nature) is
the species.
2 The taxa (categories) employed are
as follows
The species is the foundation
(plural species)
Similar species are grouped into
genera (singular- genus).
Similar genera are grouped into
families.
Similar families are grouped into
orders.
Similar orders are grouped into
classes.
Similar classes are grouped into
phyla (phylum).
Similar phyla are grouped into
kingdoms.
Other categories such as sub-species,
variety,strain, division, tribe, etc.
are employed in special circumstances.
D The scientific name of an organism is its
generic name plus its species name.
This is analogous to our system of
surnames (family names) and given
names (Christian names).
1 The generic (genus) name is always
capitalized and the species name
written with a small letter. They
BI.AQ. 15c. 7.69
-------
The System of Biological Classification
should also be underlined or printed
in italics when used in a technical
sense. For example:
Homo sapiens - (asentiens) modern man
Homo heidelbergensis - heidelberg man
Homo neanderthalis - neanderthal man
Oncorhynchus gorbuscha - pink salmon
Oncorhynchus kisutch - coho salmon
Oncorhynchus tshawytscha - chinook
salmon
Common names do not exist for most
of the smaller and less familiar
organisms. For example, if we wish
to refer to members of the genus
Gomphonema {a diatom) we must
simply use the generic name, and:
Gomphonema olivaceum
Gomphonema parvulum
Gomphonema abbreviatum
three distinct species which nave
different significances to algologists
interpreting water quality
A complete list of the various
categories to which an organism
belongs is known as its "classification"
For example, the classification of a
type of diatom and a midge larva or
"bloodworm" are shown side by side
below Their scientific names are
Gomphonema olivaceum and Chironomus
riparius.
Examples of the Classification of
an animal and a plant:
Kingdom Plantae Animalia
Phylum Chrysophyta Arthropoda
Class Bacillariophyceae Insecta
Order Pennales Diptera
Family Gomphonemaceae Chironomida
Genus Gomphonema Chironomus
Species olivaceum riparius
b These seven basic levels of
organization are often not enough
for the complete designation of
one species among thousands,
however, and so additional
echelons of terms are provided
by grouping the various categories
into "super.. . " groups and sub-
dividing them into "sub. .. " groups
Superorder, Order, Suborder, etc..
Still other category names such
as "tribe", "division"," variety
"race", "section", etc., are used
on occasion.
Additional accuracy is gained by
citing the name of the authority
who first described a species
(and the date) immediately
following the species name.
Authors are also often cited for
genera or other groups.
-------
The System of Biological Classification
d A more complete classification of
the above midge, follows-
Kingdom Animalia
Superphylum Annelid
Phylum Arthropoda
Class Insecta
Order Diptera
Suborder Nematocera
Family Chironomidae
Subfamily Chironominae
Tribe Chironomini
Genus Chironomus
Species riparius Meigen 1804
e It should be emphasized that since
all categories above the species
level are essentially human con-
cepts, there is often divergence of
opinion ui regard to how certain
organisms should be grouped.
Changes result as knowledge
grows.
f The most appropriate or correct
names too are subject to change.
The species itself, however, as
an entity in nature, is relatively
timeless and so does not change
to man's eye.
This outline was prepared by H.W. Jackson,
Chief Biologist and R. M. Sinclair, Aquatic
Biologist, National Training Center,
Water Programs Operations, EPA, Cincinnati,
OH 45268.
-------
AQUATIC ORGANISMS OF SIGNIFICANCE IN POLLUTION SURVEYS
I INTRODUCTION
A Any organism encountered in a survey is
of significance Our problem is thus not
to determine which are of significance
but rather to decide"what is the signifi-
cance of each? "
B The first step in interpretation is
recognition "The first exercise in
ecology is systematics "
C Recognition implies identification and an
understanding of general relationships
(systematics) The following outline will
thus review the general relationships of
living (as contrasted to fossil) organisms
and briefly describe the various types
II THE GENERAL RELATIONSHIPS OF
LIVING ORGANISMS
A Living organisms have long been grouped
into two kingdoms' plant and animal
Modern developments, however, have
made this simple pattern technically
untenable It has become evident that
there are as great and fundamental
differences between certain other groups
and these (two), as there are between the
traditional "plant" and "animal. " The
accompanying chart consequently shows
the fungi as a third "kingdom "
B The three groups are essentially defined
as follows on the basis of their nutritional
mechanisms (see figure
1 Plantae- photosynthetic, synthetizmg
their own organic substance from
inorganic minerals Ecologically
known as PRODUCERS.
2 Animalia ingest and digest solid
particles of organic food material
Ecologically known as CONSUMERS.
3 Fungi extracellular digestion
(enzymes secreted externally )
Food material then taken in through
cell membrane where it is metab-
olized and reduced to the mineral
condition Ecologically known as
REDUCERS.
C Each of these groups includes simple,
single-celled representatives, persisting
at lower levels on the evolutionary stems
of the higher organisms.
1 These groups span the gaps between
the higher kingdoms with a multitude
of transitional forms They are
collectively called the PROTISTA.
2 Within the protista, two principal
sub-groups can be defined on the
basis of relative complexity of
structure
a The bacteria and blue-green algae,
lacking a nuclear membrane may
be considered as the lower
protista (or Monera)
b The single-celled algae and
protozoa are best referred to as
the higher protista
D Distributed throughout these groups will
be found most of the traditional "phyla"
of classic biology.
Ill PLANTS
A The vascular plants are usually larger
and possess roots, stems and leaves.
1 Some types emerge above the surface
(emersed).
2 Submersed types typically do not
extend to the surface.
BI. AQ. 22b. 7 69
-------
Aquatic Organisms of Significance
MINERALS
Figure 1. BASIC CYCLES OF LIFE
-------
Aquatic Organisms of Significance
3 Floating types may be rooted or free-
floating.
B Algae generally smaller, more delicate,
less complex in structure, possess
chlorophyll like other green plants.
For convenience the following artificial
grouping is used in sanitary science.
1 "Blue-green algae" are typically small
and lack an organized nucleus, pigments
are dissolved in cell sap Structure
very simple.
2 "Pigmented flagellates" possess nuclei,
chloroplasts, flagellae and a red eye
spot This is an artificial group con-
taining several remotely related organ-
isms, may be green, red, brown, etc.
3 "Diatoms"have "pillbox" structure of
S1O2 - may move. Extremely common
Many minute in size, but colonial forms
may produce hair-like filaments
Golden brown in color
2 Heterotrophic bacteria are most
common. They require organic
material on which to feed.
B "True fungi" usually exhibit hyphae as the
basis of structure
V ANIMALS
A Lack chlorophyll and consequently feed on
or consume other organisms. Typically
ingest and digest their food.
B The Animal Phyla
1 PROTOZOA are single celled organisms,
many resembling algae but lacking
chlorophyll (cf illustration in "Oxygen"
lecture)
2 PORIFERA are the sponges, both marine
and freshwater representatives.
4 "Non-motile green algae" have no loco-
motor structure or ability in mature
condition Another artificial group
a Unicellular representatives may be
extremely small
b Multicellular forms may produce
great floating mats of material
IV FUNGI
Lack chlorophyll and consequently most are
dependent on other organisms They secrete
extracellular enzymes and reduce complex
organic material to simple compounds which
they can absorb directly through the cell wall.
A Schizomycetes or bacteria are typically
very small and do not have an organized
nucleus
1 Autotrophic bacteria utilize basic food
materials from inorganic substrates.
They may be photo-synthetic or
chemosynthetic.
3 CNIDARIA (= COE LENT ERA TA)
include corals, marine and fresh-
water jelly fishes, marine and
freshwater hydroids.
4 PLATYHELMTNTHES are the flat worms
such as tape worms, flukes and Planaria.
5 NEMATHELMNTHES are the round
worms and include both free-living
forms and many dangerous parasites.
6 ROTIFERS are multicellular micro-
scopic predators
7 BRYOZOA are small colonial sessile
forms, marine or freshwater
8 MOLLUSCA include snails and slugs,
clams, mussels and oysters, squids
and octopi
9 BRACHIOPODS are bivalved marine
organisms usually observed as fossils
10 ANNELIDS are the segmented worms
such as earthworms, sludge worms and
many marine species
-------
Aquatic Organisms of Significance
11 ECHINODERMS include starfish, sea
urchins and brittle stars. They are
exclusively marine.
12 CTENOPHORES, or comb jellies, are
delicate jelly-like marine organisms.
13 ARTHROPODA, the largest of all
animal phyla. They have jointed ap-
pendages and a chitinous exoskelton.
a CRUSTACEA are divided into a
cephalothorax and abdomen, and
have many pairs of appendages,
including paired antennae.
1) CLADOCERA include Daphnia
a common freshwater micro-
crustacean, swim by means of
branched antennae.
2) ANOSTRACA ( = PHYLLOPODS)
are the fairy shrimps, given
to eruptive appearances in
temporary pools.
3) COPEPODES are marine and
freshwater microcrustacea--
swim by means of unbranched
antennae.
4) OSTRACODS are like micro-
scopic "clams with legs. "
5) ISOPODS are dorsoventrally
compressed, called sowbugs.
Terrestrial and aquatic,
marine and freshwater.
6) AMPHIPODA - known as scuds,
laterally compressed. Marine
and freshwater.
7) DECAPODA - crabs, shrimp,
crayfish, lobsters, etc.
Marine and freshwater.
b INSECTA - body divided into head,
thorax and abdomen, 3 pairs of legs,
adults typically with 2 pairs of
wings and one pair of antennae.
No common marine species. Nine
of the twenty-odd orders include
species with freshwater-inhabiting
stages in their life history as follows
1) DIPTERA - two-winged flies
2) COLEOPTERA - beetles
3) EPHEMEROPTERA - may flies
4) TRICHOPTERA - caddis flies
5) PLECOPTERA - stone flies
6) ODONATA - dragon flies and
damsel flies
7) NEUROPTERA - alder flies,
Dobson flies and fish flies
8) HEMIPTERA - true bugs,
sucking insects such as water
striders, electric light bugs
and water boatman
9) LEPIDOPTERA - butterflies
and moths, includes a few
freshwater moths
c ARACHINIDA - body divided into
cephalothorax and abdomen, 4 pairs
of legs - spiders, scorpions, ticks
and mites. Few aquatic repre-
sentatives except for the freshwater
mites and tardigrades.
C CHORDATA
1 PROTOCHORDATES-primitive marine
forms such as acorn worms, sea
squirts and lancelets
2 VERTEBRATES - all animals which
have a backbone
a PISCES or fishes- including such
forms as sharks and rays,
lampreys, and higher fishes, both
marine and freshwater
b AMPHIBIA - frogs, toads, and
salamanders - marine species
rare
c REPTILA - snakes, lizards and
turtles
d MAMMALS - whales and other
warm-blooded vertebrates with
hair
e AVES - birds - warm-blooded
vertebrates with feathers
-------
Aquatic Organisms of Significance
REFERENCE
Whittaker, R.H. New Concepts of
Kingdoms of Organisms. Science
163-150-160. 1969.
This outline was prepared by H. W. Jackson
Chief Biologist, National Training Center,
Water Programs Operations, EPA, Cincinnati,
OH 45268.
-------
Aquatic Organisms of Significance
FUNGI
3/4
Sohizonyoetes - Bacteria, free living representatives
§ * >
' »
-------
Aquatic Organisms of Significance
BLUE GREEN ALGAE
3/k
Oaoillatoria spp.. filaments (triohomes) rang# from .6 to over
60/i in diameter. Ubiquitous, pollution tollerant.
Lyngbia spp.. similar to Osoillatorla but has a sheath.
A, Lvnebia aontorta. reported to be generally intollerant
of pollution ( Bc L. Mrgei.
Aphanizomenon flos-aouae
A, oolony; B,filament
toabaena flos-aauaa
A, akinete; B,heterooyst
H.W.Jaokson
-------
Aquatic Organisms of Significance
NON-MOTILE GREEN ALGAE: COCCOID
1/67 (CHLOROPHYCEAE)
Pediaatrum
Species of the Genus Scenedesmus
S. almorphuB
audatuB
Desmids
Cloaterlun
i
Staur-
aatrvun
-------
Aquatic Organisms of Significance
NON-MOTILE GREEN ALGAE: FILAMENTOUS
(Chlorophyceae)
jtydro die tyon
-------
Aquatic Organisms of Significance
PIGMENTED FLAGELLATES
3/4
A, for* of oolonyi B,o«ll la lorloa.
H.W. Jaokiea
-------
Aquatic Organisms of Significance
DIATOMS
Valva viawa
A dlsoold or oentral
diatom aaoh as
Stephanodisou a
Girdle views,
stylized to show
basic diatom
stroetar*.
£
A pennnta or navioalar
diatom suoh as
Fraglllarla
A oolony of Aatarlonella
«)
(girdle views]
rczDi
T ¥T"¥ T
A oolony of Fraglllarla
(girdle Tim)
w A
1 B
A,valve view; B,girdle
view.
ir.r*:?
Diagram showing progressive diminution in the site of oertaln
frustules tnxough stooeaaive oell generations of a diatom.
H.V. J lea on
-------
Aquatic Organisms of Significance
3/4
FREE LIVING PROTOZOA
1. Flagellated Protozoa, Class Mastigophora
Aathophvsla
Pollution tolerant
6/1
Pollution tolerant
19/t
Colony or
Pollution tolerant ,
II. Anabold Prototoe, Class Saroodlna
Dimastlgamoefra
Pollution tolerant
10-50 /i
Huolearla.reported
to be intolerant of
pollution, 45 yu.
III. Ciliated Protoroa, Class Clllopbora
Colpoda
Pollution tolerant
20-120 yu
Holophrya.reported
to be intolerant of
pollution, 35
PLATE VII
Dlfflugla
Pollution tolerant
60-500 jk
tplstylls. pollution
tolerant Colonies often
¦seroeoopio.
-------
Aquatic Organisms of Significance
PLANKTONIC PROTOZOA
Peranema trichophorum
Arcella
vulgaris
Actinosphaerium
Chaos
Tintinnidium
Codonella
Vorticella
fluviatle
-------
Aquatic Organisms of Significance
PLANKTONIC ROTIFERS
Various Forms of Keratella cochlearis
Rotaria sp
Synchaeta
pectinata
Polygarthra
vulgaris
Brachionug
quadridentata
-------
Aquatic Organisms of Significance
FREE LIVING NEMATHELMINTHES, OR ROUND WORMS
Honhyetera !j?
0
BhaMltls cf
AchromaAora
PLATE X
ejaculatory
ectal
ring
excretory pore
salivary gland
renette
intestine
testis
gland
gland
spicule
gubernaculuin
seminal vesicle
-------
Aquatic Organisms of Significance
FRESH WATER ANNELID WORMS
Phylum Annelida
anas
tube*
- mouth
Class Oligochaeta, earthworms
Ex: TubIfex , the sludgevorm
(After Liebman)
H .W .Jackson
mouth
posterior sucker disc
Class Hirudinea, leeches
(After Hegner)
anterior end
Class Polychaeta , polychaet worms
Ex.- Manayunkla, a minute, rare, tube
building vor».
(After Leidy)
-------
Aquatic Organisms of Significance
SOME MOLLUSCAN TYPES
Class: Cephalopoda*.
Squids, octopus,
cuttlefish.
Exclusively marine.
The giant squid shown
was captured in the
Atlantic in the early
ninteenth century.
(After Hegner)
liaax, Lymnaea Campeloma
a slug an air breathing mail a water breathing
snail
Class: Gaetifopodai snails and slugs. (After Buchsbanm)
Class: Pelecypodaj clans, mussels, oysters.
Locomotion of a freshwater clam, showing how foot is extended, the tip
expanded, and the animal pulled along to its own anchor. (After Bucha-
baum) H.W.Jackson
-------
Aquatic Organisms of Significance
Class CRUSTACEA
3/4
Fairy Shrimp;
Eubranohipus. Order
Phyllopoda
20m2p am
Crayfish, or ornrdad;
Caabarua. Order Oeoapoda
10-20 cm
Sow Bug* Aaallus.
Order Isopafia
10-20 a
2-5 mm
Water Flea;
Daphnla
Ordor
Cladooera
Sbttd; Hyalalia
Ordar taphipoda
10-15 ™
Fish Louse, Argulua:
a parasitio Copepod
5-6m
H.W.Jaolc son
Copspod; Cvolops. Srder Copepoda
2-5 u
-------
Aquatic Organisms of Significance
Two Winged Flies
Order DIPTERA
Adult nidge
(Chlronomld)
Eat-tailed maggot
(Iristalls)
A, adult; B,larva.
Adult sewage fly
(Peychoda)
Midge pupa
Sewage fly papa
H.W.Jackson
After various authors
-------
Aquatic Organisms of Significance
Beetles
Order COLEOPTERA
upper
lover ey
I
Whirligig beetle (Oyrinua) A. Side view
of head of adult showing divided eye;
B, Larva; C, Adult. Carnivorous.
A diving beetle (Dytlacue)
taking air at the eurface.
The riffle beetle (Paephenua);
A, adult; B. doraal tide of larra;
C, ventral elde of larva,
herbIvoroua.
Predominantly
A diving beetle (Oyblater). She div-
ing beetlee include aome of the largeat
and ooat voracloua of aquatic lnaecta.
A, larra; B,adult.
PLATE XV
Crawling water beetle;
A;adult; B.larva. Predominantly
herblvoroua.
H.W.Jackson. After
-------
Aquatic Organisms of Significance
MINOR PHYLA
Phylum Coelenterata
Hydra with bud;
extended, and contracted^
Medusa of
Cragpsdftsusta
Phylum Bryozoa
Massive colony on
stick
Statoblast
Creeping colony
on rock
Single zooid, young statoblasts in tube
-------
Aquatic Organisms of Significance
SOME PRIMITIVE FISHES
Class Agnatha, jawless fishes (lampreys and hagfishes) - Family
PETHOMYZONTIDAE, the lampreys. Lampetra aepyptera, the
Brook Lamprey A: adult, B: larva (enlarged)
,4-
Class Chondrichthyes - cartilagenous fishes (sharks, skates, rays)
Family DASYATIDAE - stingrays. Dasyatis centroura, the Roughtail Stingray
Class Osteichthyes - bony fishes - Family ACIPENSERIDAE, sturgeon.
Acipenser fulvescens, the Lake Sturgeon
Class Osteichthyes - bony fishes - Family POLYODONTIDAE, the
paddlefishes. Polyodon spathula, the Paddlefish. A:side view B:top view
Class Osteichthyes - bony fishes - Family LEPISOSTEIDAE - gars
Lepisosteus osseus, the Longnose Gar
Class Osteichthyes - bony fishes - Family AMIIDAE, bowfins
Aniia calva, the Bowfin
Reproduced with permission; Trautman, 1957.
BI.AQ. pi. 91. 6. 60
-------
TYPES OF BONY FISHES
Family CLUPEIDAE - herrings
Dorosoma cepedianum - the eastern gizzard shad
Family ANGUILLEDAE - freshwater eels
Anguilla rostrata - the American eel
Family ESOCIDAE - pikes
Esox lucius - the northern pike
Reproduced with permission; Trautman, 1957.
BI.AQ.pl. 9m. 6. 60
Family POECILIIDAE - livebearers
Gambusia affinis - the mosquitofish
Family GADDIDAE - codfishes, hakes, haddock, burbot
Lota lota - the eastern burbot
Family SCIAENIDAE - drums
Aplodinotus grunniens - the freshwater drum
-------
SECTION B
PLANTS: PRODUCERS
The producers in both aquatic and terrestrial communities are the photosynthetic
plants, autotrophic in function, which produce "fixed" energy and carbon in the
forms of their own structure, and also oxygen that consumers can then use.
Included in this section are surveys of the aquatic plant types from algae to
vascular plants with keys for identification. All studies of productivity must
take into account the four major plant life groups phytoplankton, benthic algae,
periphyton, (in part) and macrophytes. Excessive or unwanted production of
any one or combinations of these plant life groups is now a serious problem in
environmental management.
Contents of Section B
Outline No.
Types of Algae
5
Blue-Green Algae
6
Green and Other Pigmented Flagellates
7
Filamentous Green Algae
8
Coccoid Green Algae
9
Diatoms
10
Key to Algae of Importance in Water Pollution
11
Aquatic Macrophytes
12
An Artificial Key to Some Common Plants
(Freshwater, Estuarine, and Marine)
-------
TYPES OF ALGAE
INTRODUCTION
Algae in general may be defined as small
pigmented plant-like organisms of rela-
tively simple structure Actually the
size range is extreme from only a
few microns to over three hundred
feet in length Commonly observed
examples include the greenish pond
scum or frog spittle of freshwater
ponds, much of the golden brown
slime covering rocks m a trout stream,
and the great marine kelps and sea-
weeds Large freshwater forms as
Nitella and Chara or stonewort are also
included.
Algae approach ubiquity in distribution,
In addition to the commonly observed
bodies of water, certain algae also
live m such unlikely places as thermal
springs, the surface of melting snow,
on the hair of the three toed sloth m
Central America, and m conjunction
with certain fungi to form lichens
ALGAE WILL BE GROUPED FOR THE
SAKE OF CONVENIENCE INTO FOUR
GENERAL TYPES
Blue-greens (See plate Blue-Green
Algae, Cyanophyceac) This is a valid
technical group The size range is not
very great, some being so small as to
approach the size range of the bacteria
1 These are the only algae in which
the pigments are not localized in
definite bodies but dissolved through-
out the cell Blue, red, or other
pigments are present in addition to
chlorophyll thus giving the cells a
bluish green, yellow, or red color,
at least enmasse
2 The nucleus lacks a nuclear membrane
3 Tend to achieve nuisance concentrations
more frequently in the warm summer
months and in the richer -waters.
4 Vegetative reproduction, in addition
to cell division, includes the forma-
tion of "hormogones, " or short specif-
ically delimited sections of trichomes
(filaments)
5 Spores of three types are encountered
a Akmetes are usually larger, thick
walled resting spores
b Heterocysts appear like empty
cell walls, but are actually filled
with protoplasm, have occasionally
been observed to germinate
c Endospores, also called "gonidia"
or comdia, are formed by repeated
division of the protoplast within a
given cell wall Present in only a
few genera.
£> Some common examples of blue
green algae are
Anacystis (Microcystis or
Polycystis), Anabaena, Apham-
zomenon, and Qscillatoria,
B The Pigmented flagellates fin contrast
to the non-pigmented or animal-like
flagellates) are a heterogeneous
collection of motile forms from several
different algal groups (See plate
Flagellated algae)
1 There may be one, two, four, or
more flagella per cell
2 There is a well organized nucleus
3 A light-sensitive red eyespot usually
present
MIC.cla. 19a.8.69
-------
Types of Algae
4 The chlorophyll is contained in one
or more distinctive bodies called
plastids.
5 Two or more cells may be associated
in a colony
6 Non-motile life history stages may
be encountered
7 Masses of stored starch called
pyrenoid bodies are often conspicuous
8 Some examples of pigmented flag-
ellates are- Euglena, Phacus,
Chlamydomonas, Gonium, Volvox,
Peridinium, Ceratium Mallomonas,
Synura and Dinobryon
C The Non-motile green algae constitute
another heterogeneous assembly of un-
related forms (See plate Non-Motile
Green Algae)
1 Like the flagellates they have well
organized nuclei and chloroplasts.
The shape of the chloroplast is often
distinctive
2 They lack flagella or any other loco-
motor device.
3 There is extreme structural variation
among the group.
4 Some types tend to occur as a general
planktonic mass or "bloom, " often
m combinations of two or more species
Some examples are Sphaerocystis,
Pediastrum, Scenedesmus, and the
desmid Cosmarium
5 Threadlike (filamentous) green algae may
form masses or blankets, cutting off
light, and reducing water circulation.
They also add considerably to the total
mass of organic matter Some examples
of this type are Spirogyra, Hydrodictyon,
Cladophora, Oedogonium, and Chara.
D The Diatoms constitute another valid
technical group (See plate Diatoms-
Bacillariophyceae)
1 In appearance, they are geometrically
regular in shape The presence of a
brownish pigment in addition to the
chlorophyll gives them a golden to
greenish color
2 Motile forms have a distinctive
hesitating progression.
3 The most distinctive structural
feature is the two-part shell
(frustule ) composed of silicon
dioxide (glass).
a One part fits inside the other as
the two halves of a pill box, or a
petri dish.
b The surface of these shells are
sculptured with minute pits and
lines arranged with geometrical
perfection
c The view from the side is called
the "girdle view, " that from above
or below, the "valve view "
4 There are two general shapes of
diatoms, circular (centric) and
elongate (pennate) The elongate
forms may be motile, the circular
ones are not.
5 Diatoms may associate in colonies
in various ways.
6 Examples of diatoms frequently en-
countered are Stephanodiscus
Cyclotella, Asterionella, Fragilaria,
Tabellaria, Synedra, and Nitzschia
This outline was prepared by H. W. Jackson,
Chief Biologist, National Training Center,
Water Programs Operations, EPA, Cincinnati*
OH 45268.
-------
Types of Algae
KEY FOR IDENTIFICATION OF GROUPS OF FRESHWATER A LGAE
Beginning with "la" and "lb", choose one of the two contrasting
statements and follow this procedure with the "a" and "b" state-
ments of the number given at the end of the chosen statement.
Continue until the name of the algal group is given instead of
another key number.
la. Plastid (separate color body) absent, complete protoplast
pigmented, generally blue-green, iodine starch test*
negative Blue-green algae
lb. Plastid or plastids present, parts of protoplast free of some
or all pigments, generally green, brown, red, etc., but not
blue-green, iodine starch test* positive or negative 2
2a. Cell wall permanently rigid (never showing evidence of
collapse), and with regular pattern of fine markings
(striations, etc.), plastids brown to green, iodine starch test*
negative, flagella absent, wall of two essentially similar halves,
one placed over the other as a cover Diatoms
2b. Cell wall, if present, capable of sagging, wrinkling, bulging
or rigidity, depending on existing turgor pressure of cell
protoplast, regular pattern of fine markings on wall generally
absent, plastids green, red, brown, etc., iodine starch test*
positive or negative, flagella present or absent, cell wall
continuous and generally not of two parts 3
3a. Cell or colony motile, flagella present (often not readily visible),
anterior and posterior ends of cell different from one another in
contents and often in shape Flagellate algae
3b. Non-motile, true flagella absent, ends of cells often not
differentiated Green algae and associated forms
^Add one drop Lugol's (iodine) solution, diluted 1-1 with distilled water. In about 1 minute,
if positive, starch is stained blue and, later black. Other structures (such as nucleus,
plastids, cell wall) may also stain, but turn brown to yellow.
-------
Types of Algae
CMP
COMPARISON OF FOUR MAJOR GROUPS OF ALGAE
Blue-Green
Pigmented
flagellates
Greens
Diatoms
Color
Blue-Green
(Brown)
Green
Brown
Green
Brown
(Light-Green)
Location
of pigment
Throughout
cell
In plastids
In plastids
In plastids
Starch
Absent
Present or
Absent
Present
Absent
Slimy
coating
Present
Absent
in most
Absent
in most
Absent
in most
Nucleus
Absent
Present
Present
Present
Flagellum
Absent
Present
Absent
Absent
Cell Wall
Inseparable
from slimy
coating
Thin or
Absent
Semi-rigid
smooth or
with spines
Very rigid,
with regular
markings
"Eye " spot
Absent
Present
Absent
Absent
-------
BLUE-GREEN ALGAE
I WHAT ARE THE BLUE-GREEN ALGAE"?
The blue-green algae (Myxophyceae) comprise
that large group of microscopic organisms
living in aquatic or moist habitats, carrying
on photosynthesis and having differentiation
of cells which is a little more complex than
bacteria, and simpler than all of the other
plants called algae
II WHY ARE THEY CALLED BLUE-GREEN:
In addition to the green photosynthetic pigment
(chlorophyll-a) they always have a blue pig-
ment (phyocyanin-c) which tends to give the
cushions or mats they may form a blue-green
tinge. '
III WHERE ARE THE BLUE-GREENS FOUND0
Some are free floating (pelagic and planktonic),
others grow from submerged or moist soil,
rocks, wood and other objects in both fresh-
water and marine habitats.
IV WHAT ARE SOME OF THEIR GENERAL
CHARACTERISTICS'?
Some are gelatinous masses of various shapes
floating in water. Others, microscopic in
size, grow in great numbers so as to color
the water in which they live. Structurally
their cells are similar to bacteria. Their
protoplasts may be sheathed or imbedded in
gelatin, making them slimy Cells of blue-
green algae are without organized nuclei,
central vacuoles, or cilia and flagella.
No sexual reproduction is known. Asexual
reproduction may be effected by fragmentation,
in which case special separation devices are
formed (dead cells, and heterocysts). Some
species are preserved over unfavorable
periods by special spores (akinetes and endo-
spores).
V OF WHAT IMPORTANCE ARE BLUE-
GREEN ALGAE"?
They have both positive and negative economic
significance Because they can convert
radient energy into chemical energy, they
are producers forming a first link at the base
of the food chain. Because many very in-
tricate nutritional relationships exist among
the myraids of organisms it is difficult to
know the value of the blue-greens However,
people who know what the blue-greens can do
to drinking and recreational water classify
them as of negative economic importance,
because they are often nuisances when they
impart color, bad odors, and fishy tastes,
or toxins. Some of them can foul pipes
and clog filters.
VI WHEN ARE THEY MOST COMMON"?
They are widely distributed in time and space,
but tend to reach nuisance concentrations more
frequently in the late summer and in eutrophic
waters.
VII WHAT DO BLUE-GREEN ALGAE DO
FOR A LIVING"?
The pioneer-forms are of great ecological
importance because they live in habitats fre-
quented by few other forms of live, synthesiz-
ing organic substances and building substrata
that can support other kinds of life.
A Some blue-greens live in association
with other organisms as symbionts.
Still others are found in polluted
waters because they are able to
exist in habitats poor in oxygen. The
growth of these kinds of algae under
such conditions tends to make a pol-
luted condition worse
B On the other hand some species
should be promoted because they
provide oxygen and food through photo-
synthesis. The first evident product
of photosynthesis is glycogen, and
is the cause of the brown coloration
with the iodine test. Some of the
glycogen is used to produce glycopro-
teins. The gelatinous sheath is com-
posed of pectic substances, cellulose
and related compounds
BI MIC.cla 10a.8.69
-------
Blue-Green Algae
C When blue-greens mat at the surface
of the water the increased lighting
may be too strong, resulting in a
kill. At this time they may turn
from a blue-green to a yellow-green
color. Here they decompose in
mass. The resulting intermediate
products of decomposition may be
highly undesirable, because of bad
looks, four odors, bad tastes and
toxins. Under these conditions the
BOD may produce conditions not
unlike raw sewage.
may develop a great number of "pseudovac-
uoles" filled with gas. These gas bubbles make
the algae buoyant in such a way that they may
"flower" or bloom by rising to the surface
(planktonic, heaJthy blue-greens normally
possess pseudovacuoles, which are here
excepted). Soon they begin to stink because
of the odors produced from putrefaction.
The lack of dissolved oxygen during this
period may affect other organisms.
X ARE ALL BLOOMS PUTREFACTIVE ">
VIII WHAT DO BLUE-GREEN ALGAE LOOK
LIKE UNDER THE MICROSCOPE ?
A A cross section of a typical cell
would show an outside nonliving
gelatinous layer surrounding a woody
cell wall, which is bulging from
turgor pressure from the cell (plasma)
membrane, pushing the wall outward-
ly. The protoplasm, contained with-
in the plasma membrane, is divided
into two regions The peripheral
pigmented portion called chroma-
toplasm, and an inner centroplasm,
the centroplasm contains chromatins,
which is also known as in incipient
nucleus or central body, containing
chromosomes and genes. Structures
(chromatophores or plastids) con-
taining pigments have not been found
in the blue-greens The photosyn-
thetic pigments are dissolved in the
peripheral cytoplasm, which is known
as the chromatoplasm.
B A simple way to understand the cross
section would be to compare it with
a doughnut, with the hole represent-
ing the colorless central body or
incipient nucleus, which houses the
chromatoplasm, having the charac-
teristic blue-green color from its
dissolved photosynthetic pigments
IX WHAT CAUSES THESE FOUL-TO-SMELL
UNSIGHTLY BLOOMS''
When the protoplasts become sick or old they
No. Healthy blooms are produced by myraids
of cells living near the surface of the water
at times when environmental conditions are
especiall-s#favorable for them. Putrefactive
blooms are usually from masses of algae
undergoing degradation.
XI WHAT ARE SOME OF THE MAJOR
KINDS OF BLUE-GREENS'
Most species of blue-greens may be placed
into two major groups the nonfilamentous
(coccoid) forms, and the filamentous forms.
See the set of drawings following this treat-
ment to get a graphic concept of the two
groups.
XU WHAT ARE SOME OF THE MORE
DISTINCTIVE FEATURES OF BLUE-
GREENS?
A In comparing the blue-greens with
other algae it is easier to tell what
they do not possess than what they
do. They do nQt have chromatophores
or plastids, cilia, flagella, organized
nuclei, gametes, central vacuoles,
chlorophyll-b, or true starch.
B Many of the filamentous forms, es-
pecially the Oscillatoriaceae, exhibit
an unexplained movement. When the
filamentous forms are surrounded
by a gelatinous sheath the row of cells
inside is called a trichome, and the
trichome with its enclosing sheath is
called a filament. There may be more
than one trichome within a sheath.
-------
Blue-Green Algae
True branching occurs when a cell
of the series divides lengthwise and
the outer-formed cell adds cells to
form a true branch. However, two
or more trichomes within a single
sheath may be so arranged that though
they appear to be branches, their cells
actually have all divided in the same
plane, and the trichomes have pushed
out from growth to form false branch-
ing, as in Tolypothnx.
C An occasional reticulated or bubbly
appearance is referred to as pseudov-
acuolation, and en mass imparts a
pale, yellowish color to the algae.
Under low powers these vacuoles
appear dark, under higher magnifi-
cations they are reddish.
D Vegetative reproduction in addition
to cell division for the unicellular
forms, is by special kinds of frag-
mentation. This includes the for-
mation of hormogones, which are
specifically delimited sections of
trichomes, and are characteristic
of some taxonomic entities.
E Spores of three types are encountered.
1 Akinetes are usually larger, non-
motile, thick-walled resting spores.
2 Heterocysts appear like empty cell
walls, but are filled with colorless
protoplasm and have been occasion-
ally observed to germinate.
3 Endospores, also called gonidia,
are formed by a repeated division
of the protoplast within a cell wall
container.
XIII WHAT ARE SOME EXAMPLES OF BLUE -
GREEN ALGAE?
A Anacystis (Microcystis) is common
in hard waters.
1 Colonies are always free floating.
2 Their shapes may be roughly
spherical or irregular, micro-
scopic or macroscopic.
3 The gelatinous matrix may be
extremely transparent, easily
broken up on preservation.
4 They frequently contain pseudov-
acuoles.
B Anabaena is an example of a fila-
mentous form.
1 Filaments may occur singly or
in irregular colonies, and free
floating or in a delicate nucous
matrix.
2 Trichomes have practically
the same diameter throughout,
may be straight, spiral, or
irregularly contorted
3 Cells are usually spherical,
or barrel shaped, rarely cy-
lindrical and never discoid.
4 Heterocysts are usually the same
shape but are slightly larger
than the vegetative cells.
5 Akinetes are always larger than
the vegetative cells, roughly
cylindrical, and with rounded
ends.
6 It may be readily distinguished
from Nostoc by the lack of a
firm gelatinous envelope.
7 It may produce an undesirable
grassy, moldy or other odor.
C Aphanizomenon is a strictly plank-
tonic filamentous form.
1 Trichomes are relatively straight,
and laterally joined into loose
macroscopic free-floating flake-
like colonies
2 Cells are cylindrical or barrel
shaped, longer than broad.
3 Heterocysts occur within the
filament (l. e., not terminal).
4 Akinetes are cylindrical and
relatively long.
-------
Blue-Green Algae
SOME BLUE-GREEN ALGAE
I. Norvfiiamentotis ( coccoid) .Blue-Green Algae:
W m
Anacystis (Chrdioccus) X600.
Agtnenellurn
(MerismopediuiT)^..3t600 .
sheath 'V/.'I \
/j';_p^pr(»topla8ta
/ QO°cS:3CH^r - ¦
;%<; ra/
v.-.r-'
Coccochloris (Gloeocapsa) X600.
II. Filamentous blue-green algaq:
Microcystis (X600). Polrcy8ti8
(X825)
TTFTTTIB
Trichomes of Spirulina. (X600).
Phormidium (with sheath)
(X825).
: I •*.
;'I; : A. V:^
Trichomes of Arthrospira
(X6
-------
Blue-Green Algae
D
5 Often imparts grassy or nastur-
tium-like odors to water.
Oscillatoria is a large and ubiquitous
genus.
1 Filaments may occur singly or
interwoven to form mats of
indefinite extent.
2 Trichomes are unbranched, cy-
lindrical, and practically with-
out sheaths.
3 Species with narrow trichomes
have long cylindrical cells
while those with broader tri-
chomes have short broad cells.
4 No heterocysts or akinetes are
known in Oscillatoria. It re-
produces by fragmentation from
hormongonia only.
5 Live species exhibit "oscillatoria"
movements, which are oscillating.
6 Species of Oscillatoria may be
readily distinguished from
Lyngbya by the absence of a
sheath.
E Nodularia is an occasional producer
of blooms.
Trichomes are practically the
same diameter throughout.
Sheaths are usually distinct,
fairly firm, and with a single
trichome.
REFERENCES
1 Bartsch, A. F. (ed.) Environmental
Requirements of Blue-Green Algae
FWPCA. Pacific Northwest Water
Laboratory, Corvallis, Oregon.
Ill pp. 1967
2 Desikachary, T. V. Cyanophyta, Indian
Council Agric. Res New Delhi. 1959.
3 Drouet, Francis. Mxyophyceae. Chapter
5 in Edmondson. Freshwater Biology,
p. 95-114. Wiley. 1959.
4 Drouet, Francis. Revision of the Classifi-
cation of the Oscillariaceae. Monograph
15. Acad. Nat. Sci. Phil 370 pp. 1968.
5 Jackson, Daniel F. (ed.) Algae, Man, and
the Environment. Univ. Syracuse Press
554 pp. 1968.
This outline was prepared by L. G Williams,
Formerly Aquatic Biologist, Aquatic Biology
Activities, Research and Development,
Cincinnati Water Research Laboratory, FWPCA
1 Vegetative cells, heterocysts,
and even the akinetes are broader
than long.
-------
GREEN AND OTHER PIGMENTED FLAGELLATES
I INTRODUCTION
A A flagellate is a free swimming cell
(or colony) with one or more flagella.
B Motile flagellated cells occur in most
(not all) great groups of plants and animals.
C Out main concern will be with "mature"
flagellated algae.
H THE STRUCTURE OF A PIGMENTED OR
PLANT-LIKE FLAGELLATE
A There is a well organized nucleus.
B The flagellum is a long whip-like process
which acts as a propeller.
1 It has a distinctive structure.
2 There may be one or several per cell.
C The chlorophyll is contained in one or
more chloroplasts.
D Two or more cells may be associated in
a colony.
E Non-Motile Life history stages may be
encountered.
F Size is of little use in identification.
G Pyrenoid bodies are often conspicuous.
IE The Euglenophyta or Euglena -like algae
(Figures 1-4) are almost exclusively single
celled free swimming flagellates. Nutrition
may be holophytic, holozoic, or saprophytic,
even within the same species. Referred to
by zoologists as mastigophora, many animal
like forms are parasitic or commensalistic.
Food reserves of plant-like forms are as
paramylm (an insoluble carbohydrate) and
fats (do not respond to starch test). Thick
walled resting stages (cysts) are common.
"Metabolic movement" characteristics of
some genera (Euglena).
Eyespot usually present in anterior end,
rarely more than one flagellum.
A Euglena is a large genus with pronounced
metabolic movement (Figure 1).
1 Cells spindle shaped
2 Single flagellum
3 Eyespot usually present
4 Chloroplasts numerous, discoid
to band shaped
5 E. sanguinea has red pigment.
6 E. viridis generally favors water
rich in organic matter.
7 13. gracilis is less tolerant of pollution.
B Phacus cells maintain a rigid shape
(Figure 2).
1 Often flattened and twisted, with
pointed tip or tail end.
2 Cell wall (periplast) often marked
with fine ridges.
3 P. pyrum favored by polluted water.
4 P. pleuronectes relatively intolerant
of pollution.
C Trachelomonas cells surrounded by a
distinct shell (lorica) with flagellum
sticking through hole or collar (Figure 4).
1 Surface may be smooth or rough
2 Usually brown in color
3 Some species such as T. cerebea
known to clog filters
BI, MIC, cla. 6c. 3. 70
-------
Green and Other Pigmented Flagellates
D Lepocinclis has rigid naked cells with
longitudinal or spiral ridges (Figure 3).
1 Cells uncompressed, elipsoidal to oval
(in contrast to phacus)
2 Only two species with pointed tails
3 L. texta often associated with waters
of high organic content
IV The Chlorophyta or grass green algae
(Figures 5-9) are the largest and most varied
group. Non-flagellated forms predominate but
many conspicuous flagellates are included.
Food reserves are usually stored as starch
which is readily identified with iodine.
Usually two flagella of equal length are
present. More planktonic forms are included
than in any other group, predominating in the
late spring and early autumn.
The cell is typically surrounded by a definite
wall and usually has a definite shape. Cell
pigments closely resemble those of higher
plants, but some have accessory pigments
and a few forms have little or none. The
chloroplasts always have a shape charac-
teristic of the genus.
The flagellated chlorophyta are contained in
the Order Volvocales, the Volcocine algae.
All are actively motile during vegetative
phases. May be unicellular or colonial. All
have an eyespot near the base of the flagella.
Colonies may range from a simple plate
(Gonium sociale) to a complete hollow sphere
(Volvox spp ).
A Chlamydomonas is a solitary free swimming
genus (Figure 5).
1 Species range from cylindrical to
pearshaped.
2 Some species have a gelatinous sheath.
3 There are two flagella inserted close
together.
4 Generally favored by polluted waters.
B Carteria resembles Chlamydomonas very
closely except that it has four flagella
instead of two. Generally favored by
polluted water (Figure 7).
C Phacotus usually has free swimming
biflagellate cells surrounded by biconcave
envelopes resembling two clam shells.
These are usually sculptured, dark
colored, and impregnated with calcium
carbonate.
1 The eyespot ranges from anterior
to posterior.
2 Several daughter cells may be retained
within the old envelopes of the parent
cell.
3 A clean water indicator.
D Chlorogonium is a distinctive genus in
which the cell is fusiform, the tail end
pointed, and the anterior end slightly
blunt (Figure 6).
1 The two flagella only about half as
long as the cell.
2 The cell wall is rather delicate.
3 An eyespot usually present near the
anterior end.
4 Favored by pollution.
E Gonium colonies typically have 4 to 32
cells arranged in a plate (Figure 8).
1 The cells are imbedded in a gelatinous
matrix.
2 Sixteen celled colonies move through
the water with a somersault-like
motion.
3 Four and eight celled colonies swim
flagella end first.
4 Gonium pectorale is typically a
plankton form.
F Pandorina colonies range up to 32 cells,
usually roughly spherical (Figure 9).
-------
Green and Other Pigmented Flagellates
1 Cells arranged in a hollow sphere
within a gelatinous matrix.
2 Often encountered especially in hard-
water lakes, but seldom abundant.
3 P. morum may cause a faintly fishy
odor.
G Eudorina has up to 64 cells in roughly
spherical colonies.
1 The cells may be deeply imbedded in
a gelatinous matrix.
2 Common in the plankton of soft water
lakes.
3 E. elegans is widely distributed.
4 May cause faintly fishy oaor.
H Pleodorina has up to 128 cells located
near the surface of the gelatinous matrix.
It is widespread in the United States.
I Volvox rarely has less than 500 cells
per colony.
1 Central portion of the mature colony
may contain only water.
2 Daughter colonies form inside the
parent colony.
3 V. aureus imparts a fishy odor to the
water when present in abundance.
J Chlamydobotrys has "mulberry shaped"
colonies, with biflagellate cells alternately
arranged in tiers of four each.
(Spondylomorum has quadriflagellate cells),
1 There is no enveloping sheath.
2 C. stellata is favored by pollution.
V The Pyrrhophyta includes principally the
armored or dinoflagellates (Dinophyceae)
(Figures 14-16). This group is almost
exclusively flagellated and is characterized
by chromatophores which are yellow-brown
m color. Food reserves are stored as
starch or oil. Naked, holozoic, and
saprozoic representatives are found.
Both "unarmored", and "armored" forms
with chromatophores are found to ingest
solid food readily, and holozoic nutrition
may be as important as holophytic.
The great majority have walls of cellulose
consisting of a definite number of articulated
plates which may be very elaborate in
structure. There is always a groove
girdling the cell in which one flagellum
operates, the other extends backward from
the point of origin.
Most of the dino-flagellates are marine and
some are parasitic. There are six fresh
water genera of importance in this country.
A Gymnodinum species are generally naked
except for a few freshwater species.
G. brevis (marine) is a toxic form
considered to be responsible for the
"red tide" episodes in Florida and
elsewhere.
B Species of Gonyaulax (catanella and
tamarensis) are responsible for the
paralytic shellfish poisoning.
C Ceratium is distinctive in that the
anterior and posterior ends are con-
tinued as long horns (Figure 16).
1 Seasonal temperature changes have a
pronounced effect on the shape of the
cells of this species.
2 C. hirudinella in high concentration is
reported to produce a "vile stench".
-------
Green and Other Pigmented Flagellates
D Peridinium is a circular, oval, or
angular form, depending on the view
(Figure 15).
1 Cell wall is thick and heavy.
2 Plates are usually much ornamented.
3 P^ cxnctum has been charged with a
fishy odor.
VI The Division Chrysophyta contains two
classes which include flagellates, the
Xanthophyceae or Heterokontae (yellow -
green algae) and the Chrysophyceae (golden-
green algae) (Figures 10-13). The third
class, the diatoms (Bacillarieae or
Bacillariophyceae), is not flagellated.
A None of the Xanthophyceae are included
in the present discussion.
B The Chrysophyceae possess chroma -
tophores of a golden brown color, usually
without pyrenoids. Food reserves are
stored as fats and leucosin. One or two
flagella, if two, they may be of equal or
unequal length. Internal silicious cysts
may be formed. Tend to occur in
relatively pure water. Both holozoic and
holophytic types of nutrition are found.
Certain minute forms considered to be
highly sensitive to pollution.
1 Mallomonas is a solitary, free
swimming genus with one flagellum
(Figure 13).
a Covered with silicious plates, many
of which bear long silicious spines.
b Tends to inhabit clear water lakes
at moderate depths.
c M. caudata imparts a fishy odor
to the water.
2 Chrysococcus cells are minute, with
two yellowish brown chromatophores
and one flagellum.
a Droplets of stored oil present
b Lonca distinct
c C. rufesceus a clean water form
3 Chromulina has a single flagellum,
may accumulate single large granule
of leucosin at posterior end of cell
(Figure 10).
C. rosanoffii is a clean water indicator.
4 Synura is a biflagellate form growing
in radially arranged, naked colonies
(Figure 11).
a Flagella equal in length
b Cells pyriform or egg shaped
c S. uvella produces a cucumber or
muskmelon odor
5 Uroglenopsis forms free swimming
colonies of approximately spherical
biflagellate cells embedded near the
periphery of a roughly spherical
gelatinous matrix.
a Flagella are unequal in length.
b U. americana may range up to
. 5 mm in diameter, and contain
1000 or more cells.
c U. am. also causes strong fishy
odor.
6 Dinobryon may be solitary or colonial,
free floating or attached. Colonies
are arborescent (Figure 12).
a Cells attached to bottom of open
roughly cylindrical lorica or sheath.
b Two flagella of unequal length.
c Conspicuous eyespot usually present.
d Taxonomy of the group is involved.
e D. sertularia may clog filters.
f D. divergens may cause a fishy odor.
-------
Green and Other Pigmented Flagellates
(fig 1-13 from Lackey and Callaway)
Euglena
Phacus
Lepocinclis
Trachelomonas
GREEN EUGLENOIDS
0
Chlaaydononas
Gonium
Chlorogonium
9 X/
Pandorina
GREEN PHYTOMONADS
Synura
Chromulina
Dinobryon
Mallomonas
YELLOW CHRYSOMONADS
14
s+m* 70
15
Peridinium
16
Ceratium
YELLOW-BROWN DINOFLAGELLATES
-------
Green and Other Pigmented Flagellates
Flagellates
(mastigophora)
Plant Flagellates
(Phytomastigina)
Chrysomonadina
Cryptomonadina
Phytomonadina
Animal Flagellates
(z oomastigina)
RhIZOMASTIGINA
Protomonadina
Euglenoidina
Polymastigina
Figure 17 Phylogenetic Family Tree of the Flagellates
(from Calaway and Lackey)
VII There are two distinctive groups whose
systematic position is uncertain, the chloro-
monads and the cryptomonads. Only one
genus of the latter group is included here.
A Rhodomonas may range from bright red
through pale brown to olive green.
1 Cells compressed, narrow at the
posterior end
2 Two flagella of unequal length
3 R. lacustris a small form intolerant
of pollution
REFERENCES
1 Calaway, Wilson T. and Lackey, James
B. Waste Treatment Protozoa
Flagellata. Series No. 3. Univ. Fla.
140 pp. 1962.
2 Gojdics, M. The Genus Euglena.
Univ. of Wisconsin Press, Madison.
1953.
This outline was prepared by H. W. Jackson,
Chief Biologist, National Training Center,
MDS, Water Programs Operations, EPA,
Cincinnati, OH 45268.
-------
FILAMENTOUS GREEN ALGAE 1
I MANY OF THESE FORMS ARE VISIBLE
TO THE UNAIDED EYE
A They may be several inches or even a foot
or more in length. In many cases they are
not found as isolated filaments but develop
in large aggregations to form floating or
attached mats or tufts. The attached
forms are generally capable of remaining
alive after being broken away from the
substrate.
B Included in the group are some of the most
common and most conspicuous algae in
freshwater habitats. A few of them have
been given common names such as pond
silk, green felt, frog-spawn algae, and
stone worts.
E CHARACTERISTICS OF FILAMENTOUS
ALGAE
A These algae are in the form of cylindrical
cells held together as a thread ("filament"),
which may be in large clusters or growing
separately. Some are attached to rocks
or other materials while others are free.
They may be unbranched ("simple") or
branched, the tips are gradually narrowed
("attenuated") to a point. Some are
surrounded by a mucilaginous envelope.
B Each cell is a short or long cylinder with
a distinct wall. The protoplast contains
a nucleus which is generally inconspicuous.
1 The plastid or chloroplast is the
prominent structure. It contains
chlorophyll and starch centers
("pyrenoids"), and varies in size,
shape, and number per cell. It may
be pressed against the wall ("parietal")
or extend through the central axis of
the cell ("axial").
2 Clear areas of cell sap ("vacuoles") are
generally present in the cell.
1 Including a few yellow-brown and red algae.
C Specialized structures are present in
some filaments.
1 Some filaments break up into "H"
sections.
2 Apical caps are present in others.
3 Replicate end walls are present in
some.
4 Some filaments are overgrown with a
cortex.
5 Attached filaments have the basal cell
developed into a "hold fast cell"
(hapteron).
Ill REPRODUCTION MAY TAKE PLACE
BY SEVERAL METHODS
A Cell division may occur m all cells or
in certain selected ones.
B Spores called akinetes may be formed.
C Zoospores (motile) and aplanospores
(non-motile) are common.
D Fragmentation of filaments may occur.
E Many kinds reproduce sexually, often
with specialized gamete forming cells.
IV EXAMPLES OF FILAMENTOUS GREEN
ALGAE ARE-
A Unbranched forms
^Spirogyra
*Mougeotia
Zygnema
Ulothrix
Microspora
Tribonema
Desmidium
Oedogonium
*Planktonic or occasionally planktonic
BI.MIC.cla. 14b. 3. 70
-------
Filamentous Green Algae
B Branched forms
Cladophora
Pithopora
Stigeoclonium
Chaetophora
Draparnaldia
Rhizoclonium
Audouinella
Bulbochaete
Nitella
C Specialized and related forms
Schizomeris
Comsopogon
Batrachospermum
Char a
Lemanea
Vaucheria
V Habitats include the planktonic growths as
well as surface mats or blankets and benthic
attached forms on rocks in riffles of streams,
at the shoreline of lakes and reservoirs,
concrete walls, etc.
A Attached forms may break loose to
become mixed with plankton or to form
floating mats.
B Cladophora mats are a nuisance on many-
beaches on the Great Lakes.
VI IMPORTANCE OF FILAMENTOUS
GREEN ALGAE
A They may cause clogging of sand filters,
intake screens, and canals.
B They may produce tastes and odors in
water or putrid odor (also producing
HgS which damage painted surfaces) when
washed ashore around lakes and reservoirs.
C They may cause unsightly growths or
interfere with fishing and swimming in
recreation areas.
D Some are useful as indicators of water
quality in relation to pollution.
E Together with other algae, they release
oxygen required by fish, and for self-
purification of streams.
F They may produce a slime which inter-
feres with some industrial uses of water
such as in paper manufacture and in
cooling towers.
VII CLASSIFICATION
A Ulotrichaceae
Ulothrix, Microspora, Hormidium
B Cladophoraceae
Cladophora, Pithophora. Rhizoclonium
C Chaetophoraceae
Chaetophora. Stigeoclonium. Draparnaldia
D Oedogeniaceae
Oedogonium. Bulbochaete
E Schizomeridaceae
1 Schizomeris
F Ulvaceae
Enteromorpha. Mono stroma
G Zygnemataceae
Zygnema. Spirogyra. Mougeotia
H Desmidiaceae
Desmidium. Hyalotheca
I Tribonemataceae
Tribonema. Bumilleria
J Characeae
Chara. Nitella. Tolypella
-------
Filamentous Green Algae
13. GREENS, FILAMENTOUS
WtOTHHIX
^ SPlftQGYM
STfGtOCLONH*
5 60
-------
Filamentous Green Algae
VIII IDENTIFICATION
A Branching and attenuation are of primary
importance.
B Plastids: shape, location and number per
cell are essential.
C Other characteristics include grouping
of filaments, gelatinous envelope and
special features such as "H" shaped
fragments.
REFERENCES
1 Collins, F.S. 1909. The green algae
of North America. Tufts College
Studies, Scientific Series 2:79-480.
Reprinted Hafner Publ. Co., 1928
(Reprinted, 1968) Lew's Books,
San Francisco.
2 Faridi, M. A monograph of the fresh-
water species of Cladophora and
Rhizoclonium. Ph.D. Thesis.
University Microfilms, Ann Arbor.
3 Hirn, K.E. Monograph of the
Oedogoniaceae, Hafner Publ.,
New York. 1960.
4 Pal, B.P., Kundu, B.C., Sundaralmgam,
V.S., and Venkataraman, G. S.
Charophyta. Indian Coun. Agric.
Res., New Delhi. 1962.
5 Soderstrom, J. Studies in Cladophora.
Almquist, Uppsala. 1963.
6 Tilden, J. The Myxophyceae of North
America. Minn. Geol. Surv.
(Reprinted 1967, J. Cramer, Lehre,
Germany) 1910.
7 Transeau, E.N. The Zygnemataceae.
Ohio State Univ. Press. 1951.
8 Van der Hoek, C. Revision of the
European species of Cladophora.
Brill Publ, Leiden, Netherlands. 1963.
9 Wood, R.D. and Imahari, K. A revision
of the Characeae. Volume I.
Monograph (by Wood). Vol. II,
Iconograph (by Wood & Imahari). 1964.
This outline was prepared by C. M. Palmer,
Former Aquatic Biologist, In Charge,
Interference Organisms Studies, Micro-
biology Activities, Research and
Development, Cincinnati Water Research
Laboratory, FWPCA.
-------
COCCOID GREEN ALGAE
I INTRODUCTION
For the sake of convenience, the non-motile
green algae are to be discussed in two
sections: those that tend to live as relatively
discrete or free floating planktonic units,
and those that tend to grow in masses or
mats of material, often filamentous in nature,
attached or free floating.
II The green or "grass green" algae is one
of the most varied and conspicuous groups
with which we have to deal. The forms
mentioned below have been artificially grouped
for convenience according to cell shape.
Botanists would list these genera in several
different categories in the family "Chloro-
phyceae."
These algae typically have a relatively high
chlorophyll content, and the food reserves
accumulated are typically starch. Thus
these forms will usually give a typical black
or deep purple color when treated with iodine.
A Individual cells of the following genera are
perfectly round, or nearly so. The first
does not form organized colonies. In the
next two the colonies themselves tend to
be round, and in the last, the colonies are
triangular or irregular, and the cells bear
long slender spines.
1 Chlorella cells are small and spherical
to broadly elliptical. They have a
single parietal chloroplast. This is a
very large genus with an unknown
number of similar appearing species,
living in a great variety of habitats.
Although often accumulating in great
numbers, organized colonies are not
formed.
a Chlorella ellipsoides is reported to
be a common plankton form.
b Chlorella pyrenoidosa and Chlorella
vulgaris are often found in
organically enriched waters.
Indeed a dominance of Chlorella
species is considered in some
places to be an indication that a
sewage stabilization pond is func-
tioning to maximum capacity.
c Chlorella pyrenoidosa is reported
as a filter clogger in water treat-
ment plants.
2 Sphaerocystis colonies are free floating
and almost always with a perfectly
spherical, homogeneous gelatinous
envelope. Up to 32 spherical cells
may be included. Sphaerocystis
scheoeteri, the only species, is of
wide occurrence in the plankton of
lakes and reservoirs.
3 Coelastrum forms coenobial* colonies
of up to 128 cells. Generally spherical
or polygonal in shape--both cells and
colony. Cells connected by protoplasmic
processes of varying length.
Coelastrum microporum is often
reported in the plankton of water
supplies. Not surrounded by gelatinous
envelope as in Sphaerocystis.
4 Micractmium. The cells of this alga
are spherical to broadly ellipsoidal and
are usually united in irregular 4-celled
coenobes. These in turn are almost
always united with other coenobes to
form multiple associations of up to
100 or more cells. The free face of
1 Including miscellaneous yellow-brown algae.
:;'A coenobe is a colony in which the number of cells does not increase during the life of the
colony. It was established by the union of several independent swimming cells which simply
stick together and increase in size.
BI. MIC. cla. 9c. 3. 70
-------
Coccoid Green Algae
each cell in a coenobe bears from one
to seven very long slender setae or
hairs.
Micractinium pusillum. This is a
strictly planktonic genus.
B Individual cells of the following genera
are elongate. In the first two they are
relatively straight or irregular and pointed.
The next two are also long and pointed,
but bent into a tight "C" shape (one in a
gelatinous envelope, one naked). The last
one (Actinastrum) is long and straight,
but with blunt ends, and with the cells of
a coenobe attached at a point.
1 Ankistrodesmus cells are usually long
and slender, tapering to sharp point at
both ends. They may be straight,
curved, or twisted into loose aggregations.
Ankistrodesmus falcatus is often found
in the plankton in water supplies and is
considered to be one of the forms
indicative of clean water.
2 Schroederia is a solitary, free floating
alga. Cells are long and pointed at
both ends. May be bent in various ways.
Terminal points are continued as long
slender spines which may be forked and
bent back, or end as a plate. Of the
three species reported in this country,
Schroederia setigera has been reported
in water supplies.
3 Selenastrum cells are pointed at both
ends, and bent so that their tips approach
each other. They tend to occur in groups
of 4, 8, or 16, which may be associated
with other groups to form masses of a
hundred or more cells. There is no
gelatinous envelope. Selenastrum
gracile occurs in the plankton of water
supplies.
4 Kirchneriella. The cells of this genus
are generally relatively broad, tapering
to a sharp or rounded point at each end,
and the whole cell bent into a C-shape.
They usually occur in groups of four
to eight in a broad, homogeneous,
gelatinous matrix. Kirchneriella
lunaris is known principally from the
plankton.
5 Actinastrum colonies or "coenobes"
are composed of 4, 8, or 16 elongate
cells that radiate in all directions from
a common center.
Actinastrum is a widely distributed
plankton organism. There are two
species:
Actinastrum gracillimum and
Actinastrum Hantzschu differ only
in the sharpness of the taper toward
the tips of the cells. The former has
relatively little taper, and the latter,
more.
C Cells of the following genera are
associated in simple naked colonies.
The first has elongate cells arranged
with their long axes parallel (although
some cells may be curved). The last
two are flat plate-like coenobes.
Crucigenia has four-celled coenobes
while Pediastrum coenobes may be
larger, appear plate-like, and are much
more ornate.
1 Scenedesmus is a flat plate of elliptical
to double ended pointed cells arranged
with their long axes parallel. Coenobes
consist of up to 32, but usually 4 to 8
cells. The number of cells in a
coenobe may vary from mother to
daughter colony. The appearance of
cells may vary considerably with the
species.
a Scenedesmus bijuga, S. dimorphus,
and S. quadricauda are common
planktonic forms.
b Scenedesmus quadricauda is also
common in organically enriched
water, and may become dominant.
c Scenedemus abundans is reported
to impart a grassy odor to drinking
water.
-------
Coccoid Green Algae
2 Crucigenia forms free floating four-
celled coenobes that are solitary or
joined to one another to form plate-
like multiple coenobes of 16 or more
cells. The cells may be elliptical,
triangular, trapezoidal, or semi-
circular in surface view. Crucigenia
quadrata is a species often reported
from water supplies.
3 Pediastrum. Colonies are free floating
with up to 128 polygonal cells arranged
in a single plane. There may or may
not be open spaces between the cells.
The exact arrangement of the cells
seems to depend largely on the chance
distribution of the original motile
swarming zoaspores at the time the
coenobe was formed. Peripheral cells
may differ in shape from interior cells.
a Pediastrum boryanum and P. duplex
are frequently found in the plankton,
but seldom dominate.
b Pediastrum tetras has been reported
to impart a grassy odor to water
supplies.
D Cells of the following Genera are slightly
elongated.
1 Oocystis. The cells of Oocystis may
be solitary, or up to 16 cells may be
surrounded by a partially gellatmized
and greatly expanded mother cell wall.
Cells may be ellipsoidal or almost
cylindrical, cell wall thin, no spines
or other ornamentation. Oocystis
borgei, for example, is of frequent
occurrence in the plankton.
2 Dimorphococcus cells are arranged in
groups of four, and these tetrads are
united to one another in irregularly
shaped free floating colonies by the
branching remains of old mother-cell
walls. Two shapes of cell are normally
found in each tetrad (hence the name), two
longer ovate cells end to end, and a
pair of slightly shorter, C-shaped cells
on either side. Dimorphococcus
lunatus is a widely distributed plankton
organism, sometimes reported in
considerable numbers.
E A distinctive group of green algae
characterized by a median constriction
dividing the cell into two geometrically
similar halves is known generally as the
"desmids." (Closterium and Penium do
not have this construction). Each half
of the cell is known as a "semicell, "
The nucleus lies in the "isthmus. "
Extremes of ornamentation and structural
i
variety exist. Most are unicellular, but
a few are filamentous or have the cells
associated in shapeless colonies. They
are found sparingly in the plankton almost
everywhere, but predominate m acid
waters.
1 Closterium is one of the exceptional
genera without a median constriction.
The cells are elongate, attenuated
toward the tips but not sharply pointed,
usually somewhat bent.
a Closterium aciculare is a planktonic
species.
b Closterium moniliforme is reported
as a filter clogging organism.
2 Cosmarium is a large, poorly defined
genus of over 280 species, many of
which apparently intergrade with other
genera such as Staurastrum. In
general, it can be said that Cosmarium
species are relatively small, with a
length only slightly greater than the
width, and with a deep median con-
striction. Shapes of the semicells
may vary greatly. Although shallow
surface ornamentation may occur,
long spines do not occur.
a Cosmarium botrytis is reported in
plankton from water supply
reservoirs.
b Cosmarium portianum is said to
impart a grassy odor to water.
c Other species have been reported
to be sufficiently resistant to
chlorine to penetrate rapid sand
filters and occur in distribution
systems in considerable numbers.
-------
Coccoid Green Algae
3 Micrasterias is relatively common,
ornate.
4 Euastrum cells tend to be at least twice
as long as broad, with a deeply con-
stricted isthmus, and a dip or incision
at the tip of each semicell. The cell
wall may be smooth, granulate, or
spined.
Euastrum ob Ion gum is reported as a
planktonic species from water reser-
voirs. It has also been noted as
intolerant of pollution, and hence an
indicator of clean water.
5 Staurastrum is the commonest of the
desmids in the plankton of fresh waters,
the genus contains upwards of 245 species
in the United States alone. Inter-
gradation with other genera such as
Cosmarium make it a difficult group
to define. Most of the species are
radially symmetrical, and almost all
have a deeply constricted isthmus.
The cell wall may be smooth, orna-
mented, or spined in a variety of ways.
Relatively long truncated processes
extending from the cell body in
symmetrical patterns are common.
a Staurastrum polymorphum is a
typical planktonic form.
b
c Staurastrum paradoxicum causes a
grassy odor in water,
IE A type of "green" alga known as "golden
green" (Xanthophyceae) is represented in the
plankton by two genera. In these algae there
is a predominance of yellow over green pig-
ments, hence frequently imparting a yellowish
or golden tint to the cell. Reserve food
material is stored as oil and leucosin, rather
than as starch, hence giving a negative test
with iodine in most cases,
A Botryococcus braunii is a widely dis-
tributed plankton alga, though it is
rarely abundant.
1 The plant body is a free floating colony
of indefinite shape, with a cartilag-
inous and hyaline or orange-colored
envelope, surface greatly wrinkled
and folded.
2 Individual cells lie close together, in
several aggregates connected in
reticular fashion by strands of the
colonial envelope.
3 The envelope structure tends to
obscure cell structure. Considerable
deep orange colored oil may collect
within the envelope, outside of the
cells, obscuring cell structure.
B Ophiocytium capitatum like Botryococcus.
is widely distributed, but seldom abundant.
1 Both ends of cylindrical cell are
rounded, with a sharp spine extending
therefrom.
2 Many nuclei and several chloroplasts
are present.
REFERENCES
1 Palmer, C. M. Algae in Water Supplies.
Government Printing Office. PHS
Publication No. 657. 1959.
Smith, G.S. Phytoplankton of the
Inland Lakes of Wisconsin. Part I.
Bulletin No. 57, Scientific Series
No. 12. 1920.
This outline was prepared by H.W. Jackson,
Chief Biologist, National Training Center,
Water Programs Operations, EPA, Cincinnati,
OH 45268.
Staurastrum punctulatum is reported 2
as an indicator of clean water.
-------
DIATOMS
I GENERAL CHARACTERISTICS
A Diatoms have cells of very rigid form due
to the presence of silica in the wall. They
contain a brown pigment in addition to the
chlorophyll. Their walls are ornamented
with markings which have a specific pattern
for each kind.
1 The cells often are isolated but others
are in filaments or other shapes of
colonies.
2 The protoplast contains normal cell
parts, the most conspicuous being the
plastids. No starch is present.
B Cell shapes include the elongate ("pennate")
and the short cyhndric ("centric") one view
of which is circular.
1 Pennate diatoms may be symmetrical,
transversely unsymetrical, or longitudi-
nally unsymmetrical.
C Wall is formed like a box with a flanged
cover fitting over it.
1 "Valve" view is that of the top of the
cover or the bottom of the box.
2 "Girdle" view is that of the side where
flange of cover fits over the box.
3 End view is also possible for pennate
types.
D Cell markings include-
1 Raphe or false raphe extending
longitudinally.
2 Stnations which are lines of pores
extending from the area of the raphe to
the margin. Coarse ones are "costae".
3 Nodules which may be terminal and
central.
4 Internal shelves ("septae") extending
longitudinally or transversely.
II REPRODUCTION
A The common method is by cell division.
Two new half cells are formed between the
halves of the parent cell.
B Auxospores and gametes may also be
formed.
Ill EXAMPLES OF COMMON DIATOMS.
A Pennate, symmetrical
Navicula
Pinnularia
Synedra
Nitzschia
Diatoma
Fragilaria
Tabe liana
Coccone is
B Pennate, unsymmetrical
GonVphonema
Surirella
Cymbella
Achnanthes
Asterionella
Meridion
C Centric
Cyclote 11a
Stephanodiscus
Melosira
IV Habitats include fresh and salt water. Both
planktonic and attached forms occur, the latter
often are broken loose. They may be attached
by stalks or by their slimy surface.
BI. MIC. cla. 10a. 8. 69
-------
Diatoms
A Many diatoms are more abundant in late
autumn, winter, and early spring than in
the warmer season.
B The walls of dead diatoms generally remain
undecomposed and may be common m water.
Many deposits of fossil diatoms exist.
V Importance of diatoms is in part due to
their great abundance and their rigid walls.
A They are the most important group of
organisms causing clogging of sand filters.
B Several produce tastes and odors in water,
including the obnoxious fishy flavor.
C Mats of growth may cause floors or steps
of swimming pools to be slippery.
D They may be significant in determining
water quality in relation to pollution.
E They release oxygen into the water.
VI Classification. There are several thou-
sand species of diatoms. Only the most com-
mon of the freshwater forms are considered
here.
A Centrales Group
1 Representative genera.
Cyclotella
Stephanodiscus
Melosira
Rhizosolema
Biddulphia
B Pennals Group
1 Fragilanneae. The false raphe group.
Representative genera
Tabellaria
Meridion
Diatoma
Fragilaria
Synedra
Asterionella
2 Achnanthineae. Group with cells
having one false and one true raphe.
a Representative genera
Cocconeis
Achnanthes
3 Naviculmeae. True raphe group with
raphe in center of valve.
a Representative genera.
Navicula
Pinnularia
Stauroneis
Pleurosigma
Amphiprora
Gomphonema
Cymbella
Epithemia
4 Surirellineae. True raphe group with
raphe near one side of valve.
a Representative genera.
Nitzschia
Cymatopleura
Surirella
Campylodiscus
VII IDENTIFICATION OF DIATOMS
A Some genera are easily recognized by their
distinctive shape.
B Many genera and most species can be
determined only after diatoms are freed
of their contents and observed under the
high magnification of an oil immersion
lens of the compound microscope.
C Contents of the cell are generally not
used in identification. Only the char-
aracteristics of the wall are used.
-------
Diatoms
D For identification of genera, most im-
portant features include.
1 Cell shape, and form of colony
2 Raphe and false raphe
3 Striations
4 Septa
E For identification of species, measure-
ments involving the number of striae per
10 microns, the direction of the striae
and many other characteristics may be
needed.
REFERENCES
1 Boyer, C.S. The Diatomaceae of
Philadelphia and Vicinity. J. B. Lippin-
cott Co. Philadelphia. 1916. p 143.
2 Boyer, C.S. Synopsis of North America
Diatomaceae. Parts 1 ( 1927) and II
(1928). Proceedings of the Academy
of Natural Sciences. Philadelphia.
3 Elmore, C. J. The Diatoms of Nebraska.
University of Nebraska Studies. 21-
22-215. 1921.
4 Hohn, M. H. A Study of the Distribution
of Diatoms in Western New York
State. Cornell University Agricultural
Experimental Station. Memoir 308.
pp 1-39. 1951.
5 Pascher, A. Bacillariophyta (Diatomeae).
Heft 10 in Die Susswasser-Flora
Mitteleuropas, Jena. 1930. p 466.
6 Patrick, R. A Taxonomic and Ecological
Study of Some Diatoms from the
Pocono Plateau and Adjacent Regions.
Farlowia. 2.143-221. 1945.
7 Patrick, Ruth and Reimer, Charles W.
The Diatoms of the United States.
Vol. 1 Fragilariaceae, Eunotiaceae,
Achnanthaceae, Naviculaceae.
Monog. 13. Acad. Nat. Sci.
Philadelphia. 688 pp. 1966.
8 Smith, G.M. Class Bacillariophyceae.
Freshwater Algae of the United
States, pp 440-510, 2nd Edition.
McGraw Hill Book Co. New York.
1950.
9 Tiffany, L. H. and Britton, M.E. Class
Bacillariophyceae. The Algae of
Illinois, pp 214-296. University
of Chicago Press. 1952.
10 Ward, H. B. and Whipple, G.C. Class
I, Bacillariaceae (Diatoms). Fresh-
water Biology. pp 171-189. John
Wiley & Sons. New York. 1948.
11 Weber, C. I. A Guide to the Common
Diatoms at Water Pollution
Surveillance System Stations.
FWPCA. Cincinnati. 101 pp. 1966.
12 Whipple, G.C., Fair, G. M., and
Whipple, M.C. Diatomaceae.
Microscopy of Drinking Water.
Chapter 21. 4th Edition. John Wiley
& Sons. New York. 1948.
This outline was prepared by C. M. Palmer,
Former Aquatic Biologist, In Charge,
Interference Organisms Studies, Microbiology
Activities, Research and Development,
Cincinnati Water Research Laboratory,
FWPCA.
-------
II KEY TO ALGAE OF IMPORTANCE IN WATER POLLUTION
1 Plant a tube, thread. strand, ribbon, or membrane, frequently visible to the unaided eye 2
]' Plants of microscopic cells which are isolated or in irregular, spherical, or microscopic
clusters, cells not grouped into threads 123
2 (1) Plant a tube, strand, ribbon, thread, or membrane composed of cells 3
2' Plant a branching tube with continuous protoplasm, not divided into cells 120
3 (2) Plant a tube, strand, ribbon, thread, or a mat of threads 4
3' Plant a membrane of cells one cell thick (and 2 or more cells wide) 116
4 (3) Cells in isolated or clustered threads or ribbons which are only one cell thick or wide 5
4' Cells in a tube, strand, or thread all (or a part) of which is more than one cell thick or
wide 108
5 (4) Heterocysts present 6
5' Heterocysts absent 23
6 (5) Threads gradually narrowed to a point at one end 7
6' Threads same width throughout 12
7 (6) Threads as radii, in a gelatinous bead or mass 8
7' Threads not in a gelatinous bead or mass 11
8 (7) Spore (akinete) present, adjacent to the terminal heterocyst (Gloeotrichia) 9
8' No spore (akinete) present (Rivularia) 10
9 (8) Gelatinous colony a smooth bead Gloeotrichia echinulatr.
9' Gelatinous colony irregular Gloeotrichia natans
10 (8') Cells near the narrow end as long as wide Rivularia dura
10' Cells near the narrow end twice as long as wide Rivularia haematites
11 (7') Cells adjacent to heterocyst wider than heterocyst Calothrix braunn
11' Cells adjacent to heterocyst narrower than heterocyst Calothrix panetina
12 (6') Branching present 13
12' Branching absent 14
13 (12) Branches in pairs Scytonema tolypothncoides
13' Branches arising singly . Tolypothrix tenuis
14 (121) Heterocyst terminal only (Cyclindrospermum) 15
14' Hetrocysts intercalary (within the filament) 16
15 (14) Heterocyst round Cylindrospermum muscicola
15' Heterocyst elongate Cyhndrospermum stagnate
16 (14') Threads encased in a gelatinous bead or mass 17
16' Threads not encased in a definite gelatinous mass 18
17 (16) Heterocysts and vegetative cells rounded Nostoc pruniforme
17' Heterocysts and vegetative cells oblong Nostoc carneum
18 (161) Heterocysts and vegetative cells shorter than the thread width Nodulana spumigena
18' Heterocysts and vegetative cells not shorter than the thread width J9
19 (18') Heterocysts rounded (Anabaena), 20
19' Heterocysts clindnc. Aphanizomenon flos-aquae
20 (19) Cells elongate, depressed in the middle, heterocysts rare. Anabaena constncta
20' Cells rounded, heterocysts common 21
21 (201) Heterocysts with lateral extensions. Anabaena planctonica
21' Heterocysts without lateral extensions 2-2
Bl.MIC.cla. 8b. 8.69
-------
22 (2V)
22'
23 (5')
23'
24 (23)
24'
25 (23)
25'
26 (25')
26'
27 (26')
27'
28 (27)
28'
Threads 4-8y wide
Threads 8-14uwide
Branching absent
Branching (including "false" branching) present
Cell pigments distributed throughout the protoplasm
Cell pigments limited to plastids
Threads short and formed as an even spiral
Threads very long and not forming an even spiral
Several parallel threads of cells in one common sheath
One thread per sheath if present
Sheath or gelatinous matrix present
No sheath nor gelatinous matrix apparent (Oscillatona)
Anabaena fios-aquae
Anabaena circinalis
24
84
25
49
285
26
Microcoleus subtorulosus
27
28
35
Sheath distinct no gelatinous matrix between threads (Lyngbya) 29
Sheath indistinct or absent, threads interwoven with gelatinous matrix between (Phormidium)
32
29 (28) Cells rounded
29' Cells short cyhndnc
30 (29') Threads in part forming spirals
30' Threads straight or bent but not in spirals
31 (30') Maximum cell length 3 5^ , sheath thin
31' Maximum celJ length 6 5m , sheath thick
32 (28r) Ends of some threads with a rounded swollen "cap" cell
32' Ends of all threads without a "cap" cell
33 (32) End of thread (with "cap") abruptly bent
33' End of thread [with "cap") straight
34 (32*) Threads 3-5^ in width
34' Threads 5-l2M in width
35 (27') Cells very short, generally less than 1/3 the thread diameter
35' Cells generally 1/2 as long to longer than the thread diameter
36 (35) Cross walls constricted
36' Cross stalls not constricted
37 (36') Ends of thread, if mature, curved
37' Ends of thread straight
Lyngbya ocracea
30
Lyngbya lagerheimn
31
Lyngbya digueti
Lyngbya versicolor
33
34
Phormidium uncinatum
Phormidium autumnale
Phormidium inundaturn
Phormidium retzn
36
39
Oscillatona ornata
37
38
Oscillatona iimosa
38 (37)
38'
39 (35')
39'
40 (391)
40'
41 (40)
41'
42 (41')
42'
43 (40l)
43'
44 (43)
44'
Threads 10-14^ thick
Threads 16-60»» thick
Threads appearing red to purplish
Threads yellow-green to blue-green
Threads yellow-green
Threads blue-green
Cells 4-7 times as long as tne thread diameter
Cells less than 4 times as long as the thread diameter
Prominent granules ("pseudovacuoles") in center of each cell
No prominent granules in center of cells
Cells 1/2-2 times as long as the thread diameter
Cells 2-3 times as long as the thread diameter
Cel! walls between cells thick and transparent
Cell walls thin, appearing as a dark line
Oscillatoria curvicepa
Oscillatona princeps
Oscillatona rubescens
40
4]
43
Oscillatoria putrvda
42
Oscillatoria lauterbornn
Oscillatona chlonna
44
48
Oscillatona pseudogeminata
45
-------
45 (441) Ends of thread straight
45' Ends of mature threads curved
Oscillatoria agardhn
46
46 (45') Prominent granules present especially at both ends of each cell
46' Cells without prominent granules
47 (46') Cross walls constricted
47' Cross walls not constricted
Oscillatoria tenuis
47
Oscillatoria chalybea
Oscillatoria formosa
48 (43') End of thread long tapering
48' End of thread not tapering
Oscillatoria splendida
Oscillatoria amphibia
49 (24')
49'
Cells separate from one another and enclosed in a tube (Cymbella)
Cells attached lo one another as a thread or ribbon
251'
50
50 (49') Cells separating readily into discs or short cylinders their circular face showing radial
markings . 233
50' Cells either not separating readily, or if so. no circular end wall with radial markings 51
51 (50') Cells in a ribbon attached side by side or by their corners 52
51' Cells in a thread, attached end to end 56
52 (51) Numerous regularly spaced markings in the cell wall
52' Numerous markings in the cell wall absent (Scenedesmus)
53
128
53 (52) Wall markings of two types one coarse, one fine
53' Wall markings all fine (Fragilaria)
185
54
54 (531) CellB attached at middle portion only
54' Cells attached along entire length
55 (54'] Cell length 25-100i>
55' Cell length 7-25(i
56 (51') Plastid in the form of a spiral band (Spirogyra)
56' Plastid not a spiral band
57 (56) One plastid per cell
57' Two or more plastids per cell
58 (57) Threads 18-26|»wide
58' Threads 28-50»i wide
59 (58') Threads 28-40>i wide
59' Threads 40-50|J wide
60 (57') Threads 30-45|< wide, 3-4 plastids per cell
60' Threads 50-80m wide, 5-8 plastids per cell
61 (56') Plastids two per cell
61' Plastids either one or more than two per cell
62 (61) Cells with knobs or granules on the wall
62' Cells with a smooth outer wall
63 (62) Each cell with two central knobs on the wall
63' Each cell with a ring of granules near one end
64(62') Cells dense green, each plastid reaching to the wall
64' Cells light green, plastids not completely filling the cell
65 <64') Width of thread 26-32ji, maximum cell length 60|i
65' Width of thread 30-36n. maximum cell length 120n
FragiJaria crotonen&is
55
Fragilaria capucina
Fragilana construens
57
61
58
60
Spirogyra communis
59
Spirogyra varians
Spirogyra porticalis
Spirpgyra fluviatilis
Spiroeyra maiuscula
62
66
63
64
Desmidium grevilhi
Hyalotheca mucosa
Zygnema sterile
65
Zygnema insigne
Zygnema pectinatum
66 (61') Plastid a wide ribbon, passing through the cell axis (Mougeotta)
66' Plastid or plastids close to the cell wall (parietal)
67
69
-------
67 (66)
67-
68 <67')
68'
69 (66')
69'
70 (69)
70'
7) (70)
71'
72 (70)
72'
73 (69')
73'
74 (73)
74'
75 (74')
75'
76 (75)
76'
77 (7 5')
77'
78 (73')
78'
79 (78)
79'
80 (78')
80'
81 (80)
81'
82 (81)
82'
83 (80'}
83'
84 (23')
84'
Threads with occasional 'knee-ioint" bends
Threads straight
Threads 19-24^ wide, pyrenoids 4-16 per cell
Threads 20-34p wide, pyrenoids 4-10 per cell
Mougeotla genuflexa
68
Mougeotia sphaerocarpa
Mougeotia scalaris
Occasional cells with one to several transverse wall lines near one end (Oedogonium) 70
Occasional terminal transverse wall lines not present 73
Thread diameter less than 24p
Thread diameter 25n or more
Thread diameter 9-14(>
Thread diameter 14-23p
71
72
Oedogonium suecicum
Oedogonium boscn
Dwarf male plants attached to normal thread, when reproducing Oedogonium ldioandrosporum
Oedogonium g rande
No dwarf male plants produced
Cells with one plastid which has a smooth surface
Cells with several plastids or with one nodular plastid
Cells with rounded ends
Colls with flat ends (Ulothrix)
Threads lOp or less in diameter
Threads more than 10(i in diameter
Threads 5- in diamete r
Threads 6-10^ in diameter
Threads in diameter
Threads 20-60p in diameter
Iodine test for starch positive, one nodular plastid per cell
Iodine test for starch n^ative, several plastid9 per cell
Thread when broken, forming "H" shape segments
Thread when fragrrfentei, separating irregularly or between cells (Rhizoclomum)
74
78
Stichococcus bacillariB
75
76
77
Ulothrix variabilis
Ulothrix tenerrima
Ulothrix aequalis
Ulothrix Zonata
79
80
Microspora amoena
100
Side walls of cells straight, not bulging A pattern of fine lines or dots present in the wall
but often indistinct (Meloaira) ' 81
Side walls of cells slightly bulging Pattern of wall markings not present (Tnbonema) 83
Spine-like teeth at margin of end walls
No spine-like teeth present
Wall with fine granules, arranged obliquely
Wall with coarse granules, arranged parallel to sides
Plastids 2-4 per cell
Plastids more than 4 per cell
Plastids present, branching "true"
Plastids absent, branching "false"
82
Melosira varians
Melosira c renulata
Melosira granulata
Tnbonema minus
Tribonema oombycinum
85
Plectonema tomasiniana
85 (84) Branches reconnected, forming a net
85' Branches not forming a distinct net
Hydrodictyon reticulatum
86
86 (85')
86'
Each cell in a conical sheath open at the broad end I Dinobr\on)
No conical sheath around each cell
87
90
87 (86) Branches diverging, often almost at a right angle
87' Branches compace often almost parallel
Dinobryon divergens
88
88 (87') Narrow end of sheath sharp pointed
88r Narrow end of sheath blunt pointed
89
Dinobryon sertulana
-------
89 (88) Narrow end drawn out into a stalk Dinobi yon atipitatum
89' Narrow end diverging at the base Dinobryon sociale
90 (86') Short branches on the main thread in whorls of 4 or more {Nitella) 91
90' Branching commonly single or in pairs 92
91 (90) Short branches on the main thread rebranched once Nitella flcxiha
91' Short branches on the main thread rebranched two to four times Nitella gracilis
92 (90') Terminal cell eacu with a colorless spine having an abruptly swollen base (Bulbochaete) 93
92' No terminal spines with abruptly swollen bases 94
93 Vegetative cells 20-48y long Bulbochaete mirabilis
93' Vegetative cells 48-88jj long Bulbochaete insignis
94 (92') Cells red, brown, or violet Audouinclla violacea
94' Cells green 95
95 (94') threads enclosed in a gelatinous bead or mass 96
95' J breads not surrounded by a gelatinous mass 99
96 (95? Abrupt change in width from main thread to branches (Draparnaldia) 97
96' Gradual change in width from main thread to branches (Chaetophora) 98
97 (96) Branches (from the main thread) with a central, main axis Draparnaldia plumosa
97' Branches diverging and with no central main axis Draparnaldia glomerata
98 (961) End cells long-pointed, with colorless tips Chaetophora attenuata
98' End cells abruptly pointed, mostly without long colorless tips Chaetophora elegans
99 (95') Light and dense dark cells intermingled in the thread Pithophora oedogogoma
99' Most of the cells essentially alike in density 100
100 (991) Branches few in number, and short colorless Rhizoclonium hieroglyphicum
100' Branches numerous and green 101
101 (100') Terminal attenuation gradual, involving two or more cells (Stigeoclonium) 102
101' Terminal attenuation absent or abrupt, involving only one cell (Cladophora) 104
102 (101) Branches frequently in pairs 103
102' Branches mostly single StigeocIonium stagnatile
103 (102) Cells in main thread 1-2 times as long as wide Stigeoclonium lubncum
103' Cells in main thread 2-3 times as long as wide Stigeoclonium tenue
104 (lOl1) Branching often appearing forked, or in threes Cladophora aegagropila
104' Branches distinctly lateral 105
105 (104') Branches forming acute angle with main thread, thus forming clusters Cladophora glomerata
105' Branches forming wide angles with the main thread 106
106 (105') Threads crooked and bent Cladophora fracta
106' Threads straight 107
107 (I061) Branches few, seldom rebranching Cladophora insignis
107' Branches numerous, often rebranching Cladophora crispata
108 (4') Plant or tube with a tight surface layer of cells and with regularly spaced swellings (nodes)
Lemanea annulata
108' Plant not a tube that has both a tight layer of surface cells and nodes 109
109 (108') Cells spherical and loosely arranged in a gelatinous matrix Tetraspora gelatinosa
109' Cells not as loosely arranged spheres 110
110 (109') Plants branch HI
UO' Plants not branched Schizomerts leibleann
111 (110) Clustered branching 112
J1J1 Branches single 115
-------
]IZ (111) Threads embedded in gelatinous matrix (Batrachospermum)
1121 No gelantinous matrix (Chara)
113
114
113 (112)
113'
114 (UZ'l
114'
115 (111')
115'
116 (3')
116'
117 (116')
117'
118 (117')
118'
Nodal masses of branches touching one another
Nodal masses of branches separated by a narrow space
Short branches with 2 naked cells at the tip
Short branches with 3-4 naked cells at the tip
Heterocysts present, plastids absent
Heterocysts absent, plastida present
Red eye spot and two flagella present for each cell
No eye spots nor flagella present
Batrachospermum vagum
Batrachospermum momhforme
Chara globularis
Chara vulgaris
Stigonema minutum
CompBopogon coeruleus
Round to oval cells, held together by a flat gelatinous matrix (Agmenellum)
Cells not round and not enclosed in a gelatinous matrix
125
117
131
118
Cells regularly arranged to an unattached disc Number of cells 2. 4, 8, 16, 32, 64, or
128 .. . . 133'
Cells numerous, membrane attached on one surface 119
119 (118')
119'
120 (2')
120'
121 (120')
121'
122 (121')
122'
123 (]')
123'
124 (123)
124'
125 (124')
125'
126 (125)
126'
127 <126'|
127'
128 (127)
128'
129 (128")
129'
130 (129)
130'
Long hairs extending from upper surface of cells
No hairs extending from cell surfaces
Constriction at the base of every branch
No constrictions present in the tube (Vauche ria)
Chaetopeltis meealocystis
Hildenbrandia nvulariB
Dichotomosiphon tuberosus
121
Egg sac attached directly, without a stalk, to the main vegetative tube yauchena sessihs
Egg sac attached to an abrupt, short, side branch
One egg sac per branch
Two or more egg sacs per branch
Cells in colonies generally of a definite form or arrangement
Cells isolated, in pairs or in loose, irregular aggregates
Cells with many transverse rows of markings on the wall
Cells without transverse rows of markings
Cells arranged bb a layer one cell thick
Cell cluster more than one cell thick and not a flat plate
Red eye spot and two flagella present for each cell
No red eye spotB nor flagella present
Cells elongate, united side by side in 1 or 2 rows (Scenedesmus)
Cells about as long as wide
Middle cells without spines but with pointed ends
Middle cells with rounded ends
Terminal cells with spines
Terminal cells without spines
Terminal cells with two spines each
Terminal cells with three or more spines each
122
Vaucheria terrestris
Vauchena geminata
124
17 3
185
125
126
137
Gonium pectorale
127
128
131
Scenedesmus dimorphus
129
130
Scenedesmus bnuga
Scenedesmus quadricauda
Scenedesmus abundans
131 (117) Cells in regular rows, immersed in colorless matrix (Agmenellum quadriduplicatum) 132
131' CellB not immersed in colorless matrix 133
132 (131)
132'
133 (131')
133'
Cell diameter I 3 to 2 2(i
Cell diameter 3-5(i
AameneHum quadriduplicatum , tenuissima type
Agmenellum quadriduplicatum, glauca type
Cells without spines, projections, or incisions
CellB with spines, projections, or inciBions
Crucigema quadrata
134
-------
134 (133 1) Cells rounded
134' Cells angular (Pediastruml
Micractimum pusillum
135
135 (134') Numerous spaces between cells
135' Cells fitted tightly together
Prdiastrurn duplex
136
136 (135')
136'
Cell incisions deep and narrow
Cell incisions shallow and wide
Pediastrum tetras
Pcdiastrum boryanum
137 (125') Cells sharp-pointed at both ends, often arcuate
1371 Cells not sharp-pointed at both ends, not arcuate
138
141
138 (137) Cells embedded in a gelatinous matrix
138' Cells not embedded in a gelatinous matrix
Kirchneriella lunaris
139
139 (138') Cells all arcuate, arranged back to back Selenastrum g rac lie
13" Cells straight or bent in various ways, loosely arranged or twisted together
f Anki strode smusl 140
140 (139')
140'
Cells bent
Cells straight
Ankistrodesmus falcatus
Ankistrodesmus falcatus var acicularis
141 (137') Flagella present eye spots often present
141' No flagella nor eye spots present
142 (141) Each cell in a conical sheath open at the wide end IDinobrvon)
142' Individual cells not in conical sheaths
142
152
86
143
143 (142") Each cell with 1-2 long straight rods extending
143' No long straight rods extending from the cells
144 (143') CellB touching one another in a dense colony
144' Cells embedded separately in a colorless matrix
145 (144) Cells arranged radially, facing outward
145' Cells all facing in one direction
146 (145) Plastids brown, eye spot absent
146' Plastids green, eye spot present in each cell
Chrysosphaerella longispina
144
145
149
146
147
Synura uvella
Pandorina morum
147 (145')
147'
148 (147')
148'
149 (144')
149'
150 (149')
150'
151 (150')
151'
152 (141')
152'
153 (152)
153'
154 (152')
154'
155 (154)
155'
Each cell with 4 flagella
Each cell with 2 flagella (Pyrobotrys)
Eye spot in the wider (anterior) end of the cell
Eye spot in the narrower (posterior) end of the cell
Plastids brown
Plastids green
Cells 16, 32, or 64 per colony
Cells more than 100 per colony
Colony spherical, each cell with an eye spot.
Colony tubular or irregular no eye spots (Tetraspora)
Spondylomorum quaternanum
148
Pyrobotrys stellata
Pyrobotrys eracilis
Uroglenopsis americana
150
Eudorina elegans
151
Volvox aureus
109
Elongate cells, attached together at one end, arranged radially (Actinastrum)
Cells not elongate, often spherical
153
154
Cells cylindric
Cells distinctly bulging . . . .
Plastids present . . .
Plastids absent, pigment throughout each protoplast
Colonies, including the outer matrix, orange to red-brown
Matrix if any, not bright colored, cell plastids green
Actinastrum g racillimum
Actinastrum hantzschu
J55
168
Botryococcus braunn
156
-------
156 (1551) Colonies round to oval
156' Colonies not round, often irregular in form
160
157
157 (156') Straight (flat) walls between adjacent cells (Phytoconis)
157' Walls between neighboring cells rounded
278
158
158 (1571) Cells arranged as a surface layer in a large gelatinous tube (Tetraspora) 109
158' Colony not a tube, cells in irregular pattern 159
159 (158') Large cells more than twice the diameter of the small cells (Chlorococcum) . 2801
159' Large cells not more than twice the diameter of the small cells (Palme 11a) 281
160 (156) Cells touching one another, tightly grouped
160' Cells loosely grouped
161 (160') Colorless threads extend from center of colony to cells
161' No colorless threads attached to cells in colony.
162 (161) Cells rounded or straight, oval (Dictyosphaerium)
162' Cells elongate, some cells curved
163 (162) Cells rounded
163' Cells straight, oval
164 (161') Cells rounded
164' Cells oval
165 (164) One plastid per cell
165' Two to four plastids per cell
166 (165) Outer matrix divided into layers (Gloeocystis)
166' Outer matrix homogeneous
167 (166) Colonies angular
167' Colonies rounded
168 (1541) Cells equidistant from center of colony (Gomphosphaeria)
168' Cells irregularly distributed in the colony
169 (168) Cells with pseudovacuoles
169' Cells without pseudovacuoles
170 (1691) Cells 2-4(iin diameter (Gomphosphaeria lacustris)
170' Cells ovate
Coelastrum microporum
161
162
164
163
Dimorphococcus lunatus
Dictyosphaerium pulchellum
Dictyosphaerium ehrenbergianum
165
Oocystis borgei
166
Gloeococcus schroeteri
167
Sphaerocystis schroeteri
Gloeocystis planctonica
Gloeocystis gigas
169
172
Gomphospae na wichurae
170
171
Gomphosphaeria aponina
171 (170)
171'
172 (1681)
172'
Cells spherical
Cells 4-15 in diameter
Gomphosphaeria lacustris, kuetzingianum type
Gomphosphaeria lacustris, collinsn type
Cells ovid, division plane perpendicular to long axis (Coccochlorlg)
Cells rounded, or division plane perpendicular to short axis (Anacystis)
286
286'
173 (123') Cells with an abrupt median transverse groove or incision
173' Cells without an abrupt transverse median groove or incision
174
184
174 (173) Cells brown, flagella present (armored flagellates)
174' Cells green, no flagella (desmids)
17 5
178
175 (174)
175'
Cell with 3 or more long horns
Cell without more than 2 horns
Ceratium hirundinella
176
176 (175') Cell wall of very thin smooth plates
176' Cell wall of very thick rough plates (Peridinium)
Glenodlnium paluatre
177
177 (176')
177'
Ends of cell pointed
Ends of cell rounded
Pendinium wisconsinense
Peridinium cinctum
178 (174') Margin of cell with sharp pointed , deeply cut lobes or long spikes
178' Lobes, if present, with rounded ends
179
182
-------
179
179'
180
180'
181 I
181'
182
182'
183
183'
184
184'
185
185
186
186
187
187
188
188
189
189
190
190
191
191'
192
192'
193
193'
194
194'
195
195'
196
196'
197
197'
Median incision narrow, linear
Median incision wide, "V" or "U" shaped (Staurastrum)
Micrasterias truncata
180
Margin of cell with long spikes Staurastrum paradoxum
Margin of cell without long spikes 181
Ends of lobes with short spines Staurastrum polymorphum
Ends of lobes without spines Staurastrum punctulatum
Length of cell about double the width Euastrum oblongum
Length of cell one to one and one-half times the width (Cosmarium) 183
Median incision narrow linear Cosmarium botrytla
Median incision wide, "U" shaped Cosmarium portianum
Cells triangular Tetraedron muticum
Cells not triangular 185
Cells with one end distinctly different from the other 186
Cells with both ends essentially alike 225
Numerous transverse (not spiral) regularly spaced wall markings present (diatoms) 187
No transverse regularly spaced wall markings 193
Cells curved (bent) in girdle view Rhoicosphenia curvata
Cells not curved in girdle view 188
Cells with both fine and coarse transverse linea Meridion circulare
Cells with transverse lines all alike in thickness 189
Cells essentially linear to rectangular, one terminal swelling larger than the other
(Astenone 11a) 190
Cells wedge-shaped, margins sometimes wavy (Comphonema) 191
Larger terminal swelling 1-1/2 to 2 times wider than the other Asterionella formosa
Larger terminal swelling less than 1-1/2 times wider than the other Asterionella gracillima
Narrow end enlarged in valve view Comphonema geminatum
Narrow end not enlarged in valve view 192
Tip of broad end about as wide as tip of narrow end in valve view Gomphonema parvulum
Tip of broad end much wider than tip of narrow end in valve view Comphonema olivaceum
Spine present at each end of cell Schroederia setigera
No spine on both ends of cell 194
Pigments in one or more plastids 195
No plastid, pigments throughout the protoplast Entophysalis lemaniae
Cells in a conical sheath (Dinobryon) 86
Cells not in a conical sheath 196
Cell covered with scales and long spines Mallomonas caudata
Cells not covered with scales and Long spines 197
Protoplasts separated by a space from a rigid sheath (lorica) 198
No loose sheath around the cells 202
Cells compressed (flattened) Phacotus lenticularis
Cells not compressed 199
Lorica opaque, yellow to reddish or brown Trachelomonas c rebea
Lorica transparent, colorless to brownish (Chrysococcus) 200
Outer membrane (lorica) oval Chrysococcus ovalis
Outer membrane (lorica) rounded 201
-------
201 (2001) Lonca thickened around opening
201' Lorica not thickened around opening
202 (197') Front end flattened diagonally
202' Front end not flattened diagonally
203 (202) Plastids bright blue-green (Chroomonas)
203' Plastids brown, red, olive-green or yellowish
204 (203) Cell pointed at one end
204' Cell not pointed at one end
205 (203') Gullet present, furrow absent
205' Furrow present, gullet absent
206 (202') Plastids yellow-brown
206' Plastids not yellow-brown, generally green
207 (206') One plastid per cell
207' Two to several plastids per cell
208 (207) Cells tapering at each end
208' Cells rounded to oval
Chry80coccua rufeacens
.Chrysococcus maior
203
206
204
205
Chroomonas nordstetn
Chroomonas setoniensis
Cryptomonas eroaa
Rhodomonas lacustrls
Chromulina roaanoffi
207
208
211
Chlorogonium euchlorum
209
209 (2081) Two flagella per cell (Ghlamydomonas)
209' Four flagella per cell
210
Catena multifihs
210 (209) Pyrenoid angular, eye spot in front third of cell
210' Pyrenoid circular, eye spot in middle third of cell
211 (207') Two plastids per cell
211' Several plastids per cell
212 (211') Cell compressed (flattened) (Phacus)
212' Cell not compressed
Chlamydomonaa reinhardi
Chlamydomonas globosa
Cryptoglena pigra
212
213
214
213 (212) Posterior spine short, bent
213' Posterior spine long, straight
Phacus pleuronectes
Phacus longicauda
214 (212) Cell margin rigid
214' Cell margin flexible (Euglena)
215
217
215(214) Cell margin with spiral ridges Phacus pyrum
215' Cell margin without ridges, but may have spiral lines (Lepocinclis) 216
216 (215') Posterior end with an abrupt, spine-like tip
216' Posterior end rounded
Lepocinclis ovum
Lepocinclis texta
217 (214') Green plastids hidden by a red pigment in the cell
217' No red pigment except for the eye spot
Euglena eanguinea
218
218 (2171) Plastids at least 1/4 the length of the cell
218' Plastids discoid or at least shorter than 1/4 the length of the cell
219
220
219(218) Plastids two per cell .. . Euglena agilis
219' Plastids several per cell, often extending radiately from the center Euglena viridis
220 (2181) Posterior end extending as an abrupt colorless spine
220' Posterior end rounded or at least with no colorless spine
221
222
221 (220) Spiral markings very prominent and granular
221' Spiral markings fairly prominent, not granular
222 (220') Small, length 35-55(i
222' Medium to large, length 65p or more
Euglena spirogyra
Euglena oxyuris
Euglena gracilis
223
223 (222') Medium in size, length 65-200y
223' Large in size, length 250-290|J
224
Euglena ehrenbergn
-------
224 (223) Plastids with irregular edge, flagellum 2 times as long as cell Euglena polymorpha
224' Plastids with smooth edge, flagellum about 1/2 the length of the cell Cuglena deses
225 (185')
225'
226 (225)
226'
227 (226)
227'
228 (226')
228'
229 (228')
229'
230 (225)
230'
231 (230)
231'
232
232'
233 (50,
232)
233'
Cells distinctly bent (arcuate), with a spine or narrowing to a point at both ends 226
Cells not arcuate 230
Vacuole with particles showing Brownian movement at each end of cell Cells not in
c luste rs (Closterium) 227
No terminal vacuoles Cells may be in clusters or colonies 228
Cell wide, width 30-70ji
Cell long and narrow, width up to 5^
Cell with a narrow abrupt spine at each blunt end
No blunt ended cells with abrupt terminal spines
Sharp pointed ends as separate colorless spines
Sharp pointed ends as part of the green protoplast
One long spine at each end of cell
No long terminal spines
Cell gradually narrowed to the spine
Cell abruptly narrowed to the spine
A regular pattern of fine lines or dots in the wall (diatoms)
No regular pattern of fine lines or dots in the wall
Closterium momliferum
Closte rium aciculare
Ophiocytium capitatum
229
193
137
231
232
J37
Rhizosolema gracilis
233
276
Cells circular in one (valve) view, short rectangular or square in other (girdle) view 234
Cells not circular in one view 240
234 (233) Vaive surface with an inner and outer (marginal) pattern of striae (Cyclotella) 235
234' Valve surface with one continuous pattern of striae (Stephanodibcus) 238
235 (234)
235'
236 (235')
236'
237 (236)
237'
238 (234')
238'
239 (23S)
239'
240 (233')
240'
241 (240)
241'
242 (240')
242'
Cells small, 4-10n in diameter
Cells medium to large, 10-80 in diameter
Outer half of valve with two types of lines, one long, one short
Outer half of valve with radial lines all alike
Outer valve zone constituting more than 1/2 the diameter
Outer valve zone constituting more than 1/2 the diameter
Cell 4-25p in diameter
Cell 25-65(j in diameter
Cell with two transverse bands, in girdle view
Cell without two transverse bands, in girdle view
Cells flat, oval (Cocconeis)
Cells neither flat nor oval
Wall markings (striae) 18-20 in 10(i
Wall markings (striae) 23 -25 in 10M
Cyclotella glome rata
236
237
Cyclotella meneghiniana
Cyclotella bodanica
Cyclotella compta
239
Stephanodi8cus niagarae
Stephanodiacus bmderanus
Stephanodiscus hantzschu
241
242
Cocconeis pediculus
Cocconeis placentula
243
Cell sigmoid in one view
Cell not sigmoid in either round or point ended_(valve) or square ended (girdle) surface
view 244
243 (242)
243'
244 (242')
244'
245 (244)
245'
Cell sigmoid in valve surface view
Cell sigmoid in square ended (girdle) surface view
Cell longitudinally unsymmetrical in at least one view
Cell longitudinally symmetrical
Gyrosigma attenuatum
Nitzschia aciculans
245
254
Cell wall with both fine and coarse transverse lines (striae and costae)
Cell wall with fine transverse lines (striae) only
246
247
-------
246 (245) Valve face about as wide at middle as girdle face
£46* Valve face 1/2 or less as wide at middle as girdle face
247 (245) Line of pores and raphe located at edge of valve face
247' Raphe not at extreme edge of valve face
Epithemia turgida
Rhopalodia gibba
248
250
248 (247) Raphe of each valve adjacent to the same girdle surface Hantzschia amphioxys
248' Raphe of each valve adjacent to different girdle surfaces (Nitzschia) 249
249 (2481)
24^'
Cell 20-65p long
Cell 70-lSOy long
Nitzschia palea
Nitzschia linearis
250(247') Cell longitudinally unsymmetncal in valve view
250' Cell longitudinally unsymmetrical in girdle view
251
Achnanthes microcephala
251 (250)
2sr
2 52 (251')
(246)
252'
Raphe bent toward one side at the middle
Raphe a smooth curve throughout (Cymbella)
Cell only slightly unsymmetrical
Cell distinctly unsymmetrical
Amphora ovalis
252
Cymbella cesati
253
253 (2521)
2 53 1
254 (2441)
254'
Stnations distinctly cross lined, width 10-30^
Stnations indistinctly cross lined, width 5-12^
Cymbella prostrata
Cymbella ventricosa
Longitudinal line (raphe) and prominent marginal markings near both edges of valve 255
No marginal longitudinal line (raphe) nor keel, raphe or pseudoraphe median 257
255 (254) Margin of girdle face wavy
255' Margin of girdle face straight (Sunrella)
Cymatopleura solea
256
256 (255M
256'
257 (254)
2 57'
258 (257)
258'
Cell width 8-23y
Cell width 40-60^1
Sunrella ovata
Sunrella splendida
Gndle face generally in view and with two or more prominent longitudinal lines In valve
view, swollen central oval portion bounded by a line (Tabellana) 258
Girdle face with less than two prominent longitudinal lines In valve view, whole central
portion not bounded by a line
Girdle face less than 1/4 as wide as long
Girdle face more than 1/2 as wide as long
259
Tabellaria fenestrata
Tabellana flocculosa
259 (257') Valve face with both coarse and fine transverse lines
259' Valve face with transverse lines, if visible, alike in thickness
Diatoma vulga re
260
260 (259') Valve face naviculoid true raphe present
Valve face linear to linear-lanceolate, true raphe absent
261
270
261 (260) Valve face with wide transverse lines (costae) (Pinnularia)
2til' Valve face with thin transverse lines (striae)
262
263
262 (261)
? •
Cell 5-6yt broad
Cell 34-50p broad
Pinnularia subcapitata
Pinnularia nobilis
l' 3 (26D Transverse lines (striae) absent across transverse axis of valve face
Stauroneis phoenicenteron
£63' Transverse lines (striae.) present.across transverse axis of valve face 264
264 (263') Raphe strictly median (Navicula)
264' Raphe located slightly to one side
265
252
265 (264) Ends of valve face abruptly narrowed to a beak
265' Ends of valve face giadually narrowed
266 (26S1) Most of stnations strictly transverse
266' Most of stnations radial (oblique)
Navicula exigua var capitata
266
Navicula gracilis
267
-------
267 (2661) Striae distinctly composed of dots (punctae)
267' Striae essentially as continuous lines
268 (2671) Central clear area on valve face rectangular
268' Central clear area on valve face oval
Navicula lanceolata
268
Navicula graciloides
260
269 (268') Cell length 29-40p, ends slightly capitate
269' Cell length 30-120(i, ends not capitate
270 (2601) Knob at one end larger than at the other (Asterionella)
270' Terminal knobs if present equal in size (Synedra)
Navicula cryptocephala
Navicula radiosa
189
271
271 (270')
271'
272 (271')
272'
273 (272')
273'
274
274'
275 (274')
275'
Clear space (pseudonodule) in central area
No pseudonodule in central area
SideB parallel in valve view, each end with an enlarged nodule
Sides converging to the ends in valve view
Valve linear to lanceolate - linea r, 8-12 striae per 10)j
Valve narrowly linear-lanceolate, 12-18 striae per IOja
Valve 5-6>» wide
Valve 2-4 p wide
Synedra pulchella
272
Synedra capitata
273
Syned ra ulna
.274
Synedra acus
27 5
Cells up to 65 times as long as wide, central area absent to small oval
Synedra acus var radians
Cells 90-120 times as long as wide, central area rectangular
Synedra acus var augustis9ima
276 (232') Green to brown pigment in oie or more plastids
276' No pla9tids, blue and green pigments throughout protoplast
277 (276) Cells long and narrow or flat
277' Cells rounded
277
284
233
278
278 (277')
278'
279 (278')
279'
280 (279')
280'
281 C1591)
281'
282 (281')
282'
Straight, flat wall between adjacent cells in colonies
Rounded wall between adjacent cells in colonies
.Phytocoms botryoides
279
Cell either with 2 opposite wall knobs or colony of 2-4 cells surrounded by distinct mem-
brane or both 164
Cell without 2 wall knobs, colony not of 2-4 cells surrounded by distince membrane 280
Cells essentially similar in size within the colony
Cells of very different sizes within the colony
Cells embedded in an extensive gelatinous matrix
Cells with little or no gelatinous matrix around them (Chlorella)
Cells rounded
Cells ellipsoidal to ovoid
281
Chlorococcum humicola
Palmella mucosa
282
283
Chlorella ellipsoidea
283 (282) Cell 5-10p in diameter, pyrenoid indistinct
283' Cell 3-5n in diameter, pyrenoid distinct
284 (276') Cell a spiral rod
284' Cell not a spiral rod
Chlorella vulgaris
Chlorella pyrenoidosa
285
286
285 (25)
285'
286 (172)
(284')
286' (172')
Thread septate (with crosswalls)
Thread non-septate (without crosswalls)
Cells dividing in a plane at right angles to the long axis
Arthroapira lenneri
Spirulina nordstedtn
Coccochloris staRnina
Cells sperical or dividing in a plane parallel to the long axis (Anacystis)
287
287 (2861J ..Cell containing pseudovacuoles
287' Cell not containing pseudovacuoles
Anacystis cyanea
288
-------
288 (2871) Cell 2-6^ in diameter, sheath often colored
288' Cell in diameter, sheath colorless
Anacystia montana
289
289 (288') Cell 6-l2p m diameter, cells in colonies are mostly spherical
289' Cell 12-50p in diameter, cells in colonies are often angular
Anacysti8 thermalis
Anacystis dimldiata
-------
AQUATIC MACROPHYTES
I INTRODUCTION
A This non-taxonomic description of aquatic
plants includes not only the higher vascular
plants, but also the larger algae like
Chara and Lemanea and the mosses and
liverworts. Many manuals on higher
aquatic plants will also include the larger
algae and lower plants for ecological
convenience.
B Ecologically they are primary producers
as are the phytoplankton. Their role in
detrital cycles is considerable.
C The littoral zone of lakes and estuaries
are often dominated by this group while
in streams they fill important niches in
both the riffles and shallow pool areas.
Water depth determines the adjustment
of aquatic seed plants into three principal
categories.
1 Surface or floating weeds generally
grow in deeper water at the front of
(oftentimes commingling with) the
emersed weeds. The larger floating
weeds are waterlilies that may be
rooted in the mud of the bottom and bear
large leaves that float upon the surface.
Smaller types such as the duckweeds
are free-floating.
2 Emersed weeds are those that occupy
shallow water, are rooted in bottom
mud, and support foliage, seeds and
mature fruit one or more feet above
the water surface. Cattails, rushes,
and the marsh grasses are familiar
examples.
3 Submersed aquatic growths often form
a belt or zone of herbage farthest from
shore. Except for those forms that
dwell in quiet waters, they are rooted
to the bottom. Depth varies considerably
within this zone and may extend down
to the limits of effective light penetration.
D In the long-term cycle of the change in the
aquatic terrain there is a continuing
tendency for the land to encroach upon
shallow ponds and shallow areas of lakes,
decrease their size, make them more
shallow and eventually return them to dry
land. Rooted and other aquatic vegetation
plays a prominent role in this gradual
process by
1 Invading shallow water areas through
entrapment of particulate matter that
is carried into lakes and ponds. The
rooted vegetation will continue to
spread as water areas become more
shallow and the bottom mud provides
suitable anchorage for roots.
Mangrove growths in estuaries and
along seashore areas have made
swamps out of once open water areas.
2 Plants contribute also to the filling in
of lakes and estuaries through both
the precipitation of calcium carbonate
and the accumulation of their remains
upon death and decay, e.g., marl and
peat moss.
E While these lake invasions by higher
aquatic plants may sometimes be
sufficiently rapid to be recognized by
those who habitually use the lake, the
common objections to rooted vegetation
stem from their immediate interference
with recreational use such as boating and
encroachment on navigation channels and
swimming beaches.
II MACROPHYTE GROUPS
A Division Algae
The green alga Chara (and related genera
NitelLa and Tolypella) may form "weed
beds (in lakes, ponds, streams, and
estuaries) as extensive as higher plants
and grossly similar. Other algae such
as Lemanea may form dense mats in
BI. PL. lb. 3. 70
-------
Aquatic Macrophytes
riffle areas of streams or springs. In
marine waters the larger marine algae
may form dense mats.
B Division Bryophyta (Liverworts and Mosses)
These are relatively small plants which
lack flowers and conducting tissues
(xylem and phloem).
1 The Life Cycle consists of two phases:
a The leafy green gametophyte which
produces motile gametes.
b The sporophyte generation producing
spores.
2 This is a freshwater group consisting
of less than 50 genera. About one
fourth of these are liverworts.
a The liverworts (Class Hepaticae) are
small flattened green plants which
may lack stems and leaves. Riccia
is a slender, branched surface form
growing in loose clusters of flat
slender sprays. Fragments of these
are often taken in plankton samples.
Ricciocarpus is a notched oval form
about one centimeter in diameter that
is often found in the same environ-
ment.
b The true mosses (Class Musci) have
distinct leaves and stems. Sphagnum
forms extensive bogs in some areas.
In cooler streams Fontinalis may
occupy large areas^ One species of
the latter has been found resistant
to a variety of complex wastes,
including zinc.
c In swamps and along lake shores
many species pass from an aquatic
to terrestrial environment without
showing specific morphological
changes.
C Vascular Plants (Division Tracheophyta)
include the aquatic fern or fern-like plants
(Class Pteridophyta).
1 The aquatic lower tracheophytes are
primarily freshwater. The Quillworts
(Isoetaceae) are related to the club
mosses and may form numerous
rosettes on the silt-sand bottoms of
lakes.
Division Tracheophyta - This division,
like the preceding one (Bryophyta) has
the life cycle of alternation of
gametophyte and sporophyte. In this
group, however, the sporophytee
always become free living, independent
plants with possession of conducting
or vascular tissues (xylem and phloem).
The aquatic lower Tracheophyta are
primarily freshwater and only repre-
sented by a few species.
2 Included in the higher Tracheophyta
are ferns (Class Filicinae) containing
aquatic genera, Azolla, Salvinla,
Marsilea, Ceratopteris.
a Azolla and Salvinia form floating
masses sometimes interspersed
with Duckweed. The former often
forms a red carpet.
b Marsilea occupies the area between
high and low water of stream and
bayou margins.
c Ceratopteris is a floating form
often of nuisance locally.
D Vascular Plants (Division Tracheophyta,
Class Angiospermae)
1 Structural modification - all aquatic
plants are characterized by certain
specific morphological features relative
to adaptation for the aquatic habitat.
a Many heterophyllous - submerged
leaves may be narrow or incised.
Surface leaves have simple com-
pact blades.
b Root system only an anchor, with
adsorption of dissolved salts taking
place over entire plant surface,
lacking root hairs.
-------
Aquatic Macrophytes
c Stomata present only on upper side
of leaves in floating plants.
d Chloroplasts located in the epidermis.
e Flower clusters may be above water
and pollinated by wind or insects or
pollen suspended in the water and
pistils pollinated under water.
f Vegetative reproduction predominates.
Small fragments (turions or winter
buds) or root stocks (tubers) may be
sufficient for propagation.
g A variety of plants are associated
with the aquatic environment making
a clear distinction between terrestrial
and true hydrophytes difficult.
Aquatic plants have been defined by
Reid as "those whose seeds germinate
in either the water phase or the
substrate. . .and must spend part of
their life cycle in water. " This,
therefore, includes submersed as
well as emersed.
2 Classification - Around fifty families
of Angiosperms are primarily aquatic,
including thirty estuarine species.
3 Dicotyledons
a Family Nymphaceae - the water
lilies, Nuphar and Nymphaea
Watershield, Brasenia, and
Cabomba.
b Family Ceratophyllaceae - includes
one genera Ceratophyllum (Coontail
or Hornwort). The stems which are
entirely under water may sometimes
be stiff with a coat of lime.
c Family Trapaceae - the water
chestnut or caltrop, Trapa. The
sharp spined fruits are a staple
crop in parts of Asia. Colonies of
connected rosettes of this plant
quickly form surface floating mats
which may impede navigation m the
littoral region of lakes and in slow-
moving rivers. Examples - Africa,
Rumania, and Massachusetts.
d Family Haloragidaceae - includes
two genera Proserpinica and
Mynophyllum. The latter is found
up to 15 parts per 1000 salinity and
has been a serious nuisance in
Chesapeake Bay and the Tennessee
Valley. In the former area, a virus
has reduced populations as much as
95 percent.
e Podostemaceae - primarily a
tropical family which is highly
adapted to life in rushing water,
even in torrential areas over rocks
worn smooth by the water and
impenetrable by roots. This is
made possible by haptera
(attachment organs) which cement
the plant to the substrate.
Morphologically and ecologically
they bear resemblances to certain
attached algae. This group also
shares the characteristics of other
aquatic plants, but in addition
possesses quantities of silica in the
cell. Where the current in the
stream is less vigorous the
Podostemum will be replaced by
mosses and algae. Podostemum
communities are a challenge in
quantitative sampling of the
associated macromvertebrates.
f Family Lentibulariaceae -
Bladderworts, Utricularia, have
small bladders which keep the stems
afloat and also serve as traps for
the capture of small organisms.
g Family Rhizophoraceae -
Rhizophora mangle. The red man-
grove.
h Family Combretaceae -
Combretaceae - Laguncularia
racemosa. The white mangrove.
i Family Verbenaceae - Avincennia
nitida. The black mangrove
The three mangrove families listed
above are widespread on both the
East and West Coasts. They support
a unique algal vegetation, mainly of
certain red algal genera.
-------
Aquatic Macrophytes
4 Monocotyledons
a Family Butomaceae - Butomus
umbellatus is Eurasian in origin,
and has spread rapidly m the
St. Lawrence River Basin and
around the Great Lakes.
b Family Hydrocharitaceae - includes
the well known Elodea ( = Ana char is)
Hydrilla, Egeria, and Vallisneria
plus two marine genera Halophila
and Thalassia (T. testudinum,
Turtle Grass.) The former fresh
water genera also extend into
brackish water.
c The water plantain family
(Alismaceae) is a large group of
emersed marsh plants and sub-
mersed aquatics. Sagittaria
subulata is found in both fresh and
estuarme areas. In the latter it
matures without much change in
shape or size of leaves (ribbon leaf
form) and the plant and flowers may
be under water at high tide.
d Family Zosteraceae
1) Zostera marina or Eel Grass is
found on both coasts from Alaska
to California and Hudson Bay to
"North Carolina. In the early
thirties, there were great
mortalities of Eel Grass over
extensive areas, particularly the
East Coast. Since that time there
has been considerable recovery
in the east.
2) Phyllospadix scouleri or Surf
Grass is found from British
Columbia to California. The
"sea grasses" include the above
four genera and two others
(discussed in the families
Hydrocharitaceae and
Zannichelliaceae. They are
stenohalme and probably do not
occur where salinities are below
25 7oo for considerable periods.
The remaining genera of this
family are primarily fresh
water although many have species
extending into brackish water.
e Family Potamogetonaceae
Potamogeton includes over forty
species and is recognized generally
as a difficult group to key out.
f Family Ruppiaceae
Ruppia maritima or Widgeon Grass
is found from alkali to fresh water
and salt to fresh coastal water.
g Family Zannichelliaceae
1) Zannichellia palustris is found
from fresh water and fresh to
brackish coastal water.
2) Cymodoceae - Syringodium
filiforme (= Cymodocea
manatorum) or Manatee Grass
is found from Texas to Florida.
The leaves are round in cross-
section and flowers are common.
3) Halodule wrightii (= Diplanthera)
or Shoal Grass is found in North
Carolina and from Texas to
Florida. Leaves are flat in
cross-section and have a three
pointed tip.
h Family Najadaceae - includes the
one genus Naia^, approximately
35 species. Southern Naiad has
long been one of the most trouble-
some submersed aquatic weeds in
Florida. It infests irrigation and
drainage canals.
i Family Pontederiaceae - includes
Eichornia crassipes, waterhyacinth.
It is a native of tropical America
and was probably introduced in the
United States as an ornamental. As
an escapee it has become an
exceedingly troublesome species
by clogging waterways of the
Southern States. Its attractive
-------
Aquatic Macrophytes
blue-purple flowers and charac- III
teristic bulbous leaf stem with
rounded leaf blade make it easy to
identify. The plant is usually found A
floating on the surface of ponds and
quiet streams and growing on mud-
banks. This plant spreads
vegetatively by horizontal stem
growth and rooting at the nodes to B
produce new plants that develop into
mats covering large areas. The
capsulelike fruits contain many
seeds that provide for extensive
spread of the species in suitable
climates.
TYPICAL AQUATIC PLANT
COMMUNITIES
Ponds and lakes not only have a charac-
teristic plant zonation, but over a period
of years exhibit a distinct succession of
aquatic plant communities.
Intertidal salt marshes are so distinctive
that in one state,regulations preventing
the disruption of the ecology of salt
marshes, are defined on the basis of the
specific plants involved. A typical
estuarine shore (from seaward in) might
have successive zones dominated by
j Family Acanthaceae - Dianthera
(= Justicia) or Water Willow forms
large beds at the margins of streams.
k Marsh and shore zones are inhabited
by Bulrushes or Sedges (Family
Cyperaceae), Rushes (Family
Juncaceae), and Grasses
(Gramineae). The latter includes
wild rice (Zizania)and Cord Grass
(Spartina).
1 Family Amaranthaceae - includes
Alternanthera philoxeroides or
Alligator Weed.
m Family Lemnaceae - The Duck weed
family includes the smallest known
flowering plants, all of which are
free floating. Wolffia and Wolffiella
are without roots and Lemna and
Spirodela both possess them.
n Family Typhaceae - one genus
Typha. Cattail or tule.
o Family Iridaceae - Iris pseudacoris
is another European plant becoming
widely established in North America.
p Family Araceae - Acorus calamus,
sweet flag, is an Eurasian plant
which has become widely established
in North America.
1 Suaeda (alkali seepweed)
2 Spartina (cord grass)
3 Halimione
4 Puccinellia (alkaligrass) and others
5 Juncus (rush)
C Mangrove vegetation replaces salt marshes
in tropical and subtropical regions. Two
most widespread genera are-
1 Rhizophora, the red mangrove
occupies the outer pioneer zone.
Roots are borne on downward curving
branches or rhizophores. The
resulting tangle reduces tidal currents
and promotes deposition of solids.
2 Avicennia, the black mangrove,
usually forms a shoreward zone.
It depends on aerial roots, which
emerge from the ground a short
distance from the tree, rather than
prop roots.
3 Examples have been given of open
shoal areas being transformed into
thick swamp forest in some 30 years
through this process.
-------
Aquatic Macrophytes
D Sea grass communities, whether composed
of Zostera (eel grass) in Northern waters
or Thalassia (turtle grass) or Cymodocea
(Manatee grass) in the tropics, are
recognizable as an entity wherever found.
A soft substratum near low water is
required, and each has a similar distinct
assemblage of molluscs, crustacenas,
and other invertebrates.
1 The destruction of this community by
bulkheadmg and filling has far-reaching
effects on the stability of such bottom
areas.
2 Destruction of these plants by disease,
such as the epidemics which decimated
the Zostera eel grass community in the
thirties created both erosion problems
and also brought about far-reaching
biotic community changes which only
became reestablished with the return
of the eel grass.
3 Blackburn, Robert D., White, P.E. and
Weldon, L. W. Ecology of Submersed
Weeds in South Florida Canals.
Weed Science 16 (2) 261-266. 19G8.
4 Dawson, Elmer Y. Marine Botany.
Holt-Rhinehart & Winston, New York,
pp. 371. 1966.
5 Holm, L. G., Weldon, L.W. and
Blackburn, R.D, Aquatic Weeds.
Science 166*699-709. 1969.
(An excellent summary and review of
"the rampart growth of weeds has
become one of the symptoms of our
failure to manage our resources".)
6 Kormandy, Edward J. Comparative
Ecology of Sandspit Ponds. Am. Mid.
Nat. 82:28-61. 1969
7 Lawrence, J.M. Aquatic Herbicide
Data. USDA Handbook No. 231,
pp. 133. 1962.
IV In comparison to other living organisms
the aquatic plants or macrophytes have been
seriously neglected. Man's increased
activities have favored some plants (which
we call "weeds") through eutrophication and
diminished others (which we call a valuable
resource loss) through dredging and filling
operations. These events demand
reassessment of knowledge about the ecology
of these plants and increased contributions
toward increasing that information.
ECOLOGICAL REFERENCES
1 Bayley, Suzanne, Rubin, Harvey, and
Southwick, Charles H. Recent Decline
in the Distribution and Abundance of
Eurasian Milfoil in Chesapeake Bay.
Chesapeake Science 9 (3):173-181.
1968.
8 Mackenthun, Kenneth M. A Review of
Algae, Lake Weeds, and Nutrients.
J.WPC Fed. 34.1077-1085. 1962.
9 Mackenthun, Kenneth M. Nitrogen and
Phosphorus in Water. An Annotated
Selected Bibliography of Their
Biological Effects, USPHS Pub. No.
1305, pp. 111. 1965.
10 Mackenthun, Kenneth M, and Ingram,
William M. Biological Associated
Problems in Freshwater Environ-
ments, FWPCA, pp. 287. 1967.
11 Neel, Joe Kendall. Seasonal Succession
of Benthic Algae and their Macro-
invertebrate Residents in a Headwater
Limestone Stream, Jour. Water Poll.
Cont. Fed., 40 (2) Part 2. R10-30.
1968.
2 Bickel, David. The Role of Aquatic
Plants and Submerged Structures in the
Ecology of a Freshwater Pulmonate
Snail. Physa Integra Hald.
Sterkiana 18 17-20. 1965.
12 Nelson, Daniel J. and Scott, Donald C.
Role of Detritus in the Productivity
of a Rock-Outcrop Community in a
Piedmont Stream, Limn, and Ocean
7(3):396-413. 1962.
-------
Aquatic Macrophytes
6 Fassett, N. C. A Manual of Aquatic
Plants (with Revision Appendix by
Eugene C. Ogden). University of
Wisconsin Press, Madison, 405 pp
1960.
13 Peltier, W.H. and Welch, E.B.
Factors Affecting Growth Rooted
Aquatic Plants, TVA, Chattanooga,
Tennessee, pp. 45. 1968.
14 Rawls, Charles K., Jr. Reefoot Lake
Waterfowl Research, Term. Game
&. Fish Comm., pp. 80. 1954.
15 Smith, Gordon E. and Isom, Billy G.
Investigation of Effects of Large-
Scale Applications on 2, 4-D on
Aquatic Fauna and Water Quality,
Pest. Monit. Journal 1 (3).16-21. 1967.
16 Westlake, D. F. The Biology of Aquatic
Weeds m Relation to their Management.
Proc. 9th Brit. Weed Conf. pp. 372-
381. 1968.
17 Zeiger, C. F. Biological Control of
Alligatorweed with Agasicles n. sp.
in Florida, Hyacinth Cont. Jour.,
6:31-34. 1967.
IDENTIFICATION REFERENCES
1 Arber, Agnes. Water Plants, Wheldon
and Wesley, Lt. and Hafner Pub. Co.,
New York, pp. 436. 1963.
2 Blackburn, Robert D. and Weldon, Lyle
W. Eurasian Watermilfoil-Florida's
new underwater menace. Hyacinth
Cont. Jour. 6.15-18. 1967.
3 Conard, Henry S. How to Know the
Mosses, Wm. C. Brown, Dubuque,
Iowa, pp. 166. 1944.
4 Dawes, Clinton J. Marine Algae in the
vicinity of Tampa Bay, University S.
Florida. 1967. (includes Angiosperms)
5 Eyles, C.E. and Robertson, J. L.
A Guide and Key to the Aquatic Plants
of the Southeastern United States,
USPHS Bull. No. 286, 151 pp. 1944.
(Reprinted as Circular 158, U.S. Bur.
Sport Fisheries and Wildlife, 1963).
7 Fernald, M. L. Gray's Manual of
Botany. 8th ed. Amer. Book Co.
1632 pp. 1950.
8 Hotchkiss, N. Pondweeds and Pondweed-
like Plants of Eastern North America,
U. S. Fish and Wildlife Service,
Circular 187, pp. 1-30. 1964.
9 Hotchkiss, N. Bulrushes and Bulrush-
like Plants of Eastern North America,
U. S. Fish and Wildlife Service,
Circular 221, pp. 1-19. 1965.
10 Hotchkiss, N. Underwater and Floating-
Leaved Plants of the U. S. and Canada,
U. S. Fish and Wildlife Service,
Resource Publ. No. 44, pp. 124. 1967.
11 Humm, H.J. Seagrasses of the Northern
Gulf Coast, Bull. Mar. Sc. Gulf &
Caribbean 6 (3)- 305-308.
12 Muenscher, W. C. Aquatic Plants of the
United States, Comstock Publishing
Company, Ithaca, New York, 374 pp.
1944.
13 Otto, N. E. and Bart ley, T.R. Aquatic
Pests on Irrigation Systems,
Identification Guide, U.S. Bur. of
Reclamation Water Resources.Tech.
Publ., pp. 72. 1965.
14 Stewart, Albert W., Dennis, LaRue J.,
and Gilkey, Helen M. Aquatic Plants
of the Pacific Northwest. Oregon
State Monographs, Corvallis,
Second Edition, 261 pp. 1963.
15 Weldon, L. W., Blackburn, R.D., and
Harrison, D. S. Common Aquatic
Weeds, USDA Agriculture Handbook
No. 352, 43 pp. 1969.
-------
Aquatic Macrophytes
16 Winterringer, Glen S. and Lopinot, Alvin
C. Aquatic Plants of Illinois, Illinois
State Museum Popular Science Series,
Vol. VI, pp. 142. 1966.
17 Correll, Donovan S. and Correll, Helen B.
Aquatic and Wetland Plants of Southwestern
United States. Water Pollution Control
Research Series. 16030 DNL 01/72.
Environmental Protection Agency
Research and Monitoring, Wash., D. C.
1777pp. 1972.
This outline was prepared by R. M. Sinclair,
Aquatic Biologist. National Training Center,
Water Programs Operations, EPA, Cincinnati,
OH 45268.
-------
AN ARTIFICIAL KEY TO SOME COMMON AQUATIC PLANTS
{Freshwater, Estuarine, and Marine)
I Specific identification of aquatic plants
often requires microscopic examination of
parts which may not always be available to
the investigator (for example, fruiting bodies
and/or seeds which are seasonal or produced
only rarely).
Because of this, some groups are more
difficult to identify. Potamogeton is a varied
and difficult genus, although Ogden's Key
gives help where only vegetative parts are
available. Najas and Myriophyllum are often
difficult.
In the Hydrocharitaceae, Hydrilla, Egeria,
and Elodea (Anacharis is an obsolete name
for the latter two) are sometimes confused,
and the serious student may need to turn to
the specialist in this group (see Blackburn
et al for a discussion of this problem.
For a detailed list of references on classification
see Sculthorpe (p. 16-20) where these are
listed by family. The following key will aid
the common plant groups. It is divided as
follows.
A) Plants floating on water surface.
B) Plants submersed beneath water surface.
C) Plants erect and emersed, rooted to the
substratum and extending upward out
of water.
D) Submersed "sea grasses".
Manuals with descriptive comments and
figures for specific identification are listed
in part under references.
To use the key, select the proper group
(A, B, C, D) and read the description in the
first couplet. The description that best fits
the unknown specimen will indicate either the
plant group or genus to which the specimen
may belong or an additional couplet, in which
case the process is repeated until the descrip-
tion for a particular plant or genus best fits
the unidentified specimen. An asterisk *
indicates that distribution may include
brackish or salt water. Common names
used here (with few exceptions) are those
of: Subcommittee on Standardization of
Common and Botanical Names of Weeds,
reprinted from Weeds 14 <4>:347-386 (1966).
A PLANTS FLOATING ON WATER SURFACE
1. Plants very small, seldom 2
over a centimeter along
any dimension.
1. Plants large; usually 8
measuring at least 0. 5
decimeter along some
dimension.
2. Plant body dichotomously 3
2-lobed, or repeatedly
dichotomously branched.
2. Plant body not dichoto- 4
mously branched and if
2-lobed not equally so.
3. Divisions of plant body fine Riccia
and many, plant body usuallyfluitans L.
floating below surface.
3. Divisions of plant body
coarse, 2-lobed, floating
on surface.
4. Plants floating on
surface.
4. Plants floating just
below water surface;
plant body made up of
a clump of short
filaments.
5. Plant body of small over-
lapping scales.
5. Plant body simple or
compound, made up of
rounded floating
leaves.
Ricciocarpus
natans L.
Wolffiella
floridana Sm.
Azolla
BI. PL, 2b. 2.70
-------
An Artificial Key to Some Common Aquatic Plants
6. Plants simple, extremely
minute, appearing as
grains on surface of
water.
6. Plants compound, made
up of several rounded-
oblong disk-like bodies,
floating on surface of
water.
7. Plant body inconspicuously
nerved, rootlets 1 per disk.
7. Plant body conspicuously
nerved, rootlets 2 to
several per disk.
8. Leaves broad and blade-
like, sometimes inflated
near base.
8. Leaves (plant body in
Riccia) narrow or finely
divided.
9. Leaves large and dilated
with inflated petioles.
9. Leaves normally expanded.
10. Leaves bearing plantlets
around the margin.
10. Leaves not bearing
plantlets.
11. Floating-leafed plants with
leaves attached to the
bottom by a bare unbranched
stem of varying length.
11. Roots usually suspended
free in the water, with no
connection to lake bottom;
capable of drifting.
12. Leaves wide to the base,
without petioles.
12. Leaves mostly differ-
entiated into blades
and petioles.
watermeal 13. Plant bociy repeatedly Riccia
Wolffia dichotomously branched, fluitans L.
13. Leaves not dichoto- 14
mously branched.
7
14. Leaves entire not Elodea
dissected or bearing
bladders, and
whorled.
star duckweed
Lemna
14. Leaves dissected and
bearing bladders.
bladderwort
Utricularia
giant duckweed 15. Stem attached to middle 16
Spirodela
polyrhiza L.
of leaf.
15. Stem attached at the
summit of a deep notch
in the leaf.
17
12
waterhyacinth
Eichhornia
crassipes Mart.
11
floating fern
Ceratopteris
11
16. Leaves oval, not
more than 3 inches
wide, with supple
stem attached to the
middle of the leaf.
16. Circular leaf with a
long, fairly rigid
stem attached to the
middle of the leaf,
leaves 6 inches or
more wide some-
times supported by
the stem above the
water level.
watershield,
Brasenia schreberi
Gmelin
american lotus
Nelumbo
15 17. Circular or heart- yellow pond lily
shaped leaf with the Nuphar
veins radiating from
the mid-rib nearly to
the margin without
12 forking, floating yellow
flowers.
17. Circular leaf with much- *white water lily
forked veins radiating to Nymphaea
water lettuce the margin, white, or
Pistia pink floating flowers.
stratiotes L.
frogbit
Limnobium
spongia Base.
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An Artificial Key to Some Common Aquatic Plants
PLANTS SUBMERSED BENEATH WATER
SURFACE
1. Plant body made up of
stems bearing whorled,
smooth, brittle branches,
easily snapped with a
slight pressure, plants
with a musky odor, no
roots, often with a limy
encrustation.
1. Plant body not as
described.
2. Plant attached to sub-
strate by holdfasts or
haptera, not rooted.
2. Plant not so attached.
3. Plant attached to stones
by haptera (roots not
attached) and tough
stems forming a dull-
green tangle.
3. Plants single shoots
attached to substrate
by holdfasts.
4. Without true roots,
flowers or vascular
bundles.
4. Usually with true
roots and with
vascular bundles.
5. Leaves without midrib,
arranged in two opposite
rows, usually with a
part folded under.
5. Leaves most often with
midrib, usually arranged
equally around the stem,
not curved under.
*green alga,
muskgrass,
Chara
6. Submerged leaves bladderwort
bearing small Utricularia
bladders, leaves
irregularly forked.
6. Submerged leaves 7
not bladder bearing.
7. Submerged leaves com- 8
pound, made up of
narrow segments or
leaflets.
riverweed,
Podostemum
ceratophyllum
red alga
Lemanea
Division Bryophyta,
Mosses and
Liverworts
5
Division
Angiospermae.
True flowering
plants
6
Order Junger-
manmales,
Porella
Order Musci,
Leptodictyum,
Fontinalis, and
Fissidens
7. Submerged leaves
simple, made up of
a single narrow blade.
8. Submerged leaves
with one central
axis, leaves feather-
like, branches in
whorls about the
stem, stems
usually very lax.
8. Submerged leaves
irregularly forking.
9. Submerged leaves
singly and alternately
or irregularly borne;
leaves many branches,
irregularly forked and
appearing as tufts of
numerous thread-like
projections attached
to the center stem.
9. Submerged leaves
borne opposite each
other on stem or
whorled.
10. Leaves stalked, fan-
like, extending from
opposite sides of the
stem, leaflets not
toothed.
10, Stems with whorls of
stiff, forked leaves,
leaflets with toothed
or serrated margins
(small barbs)onone
side; plant without
true roots.
11
*water milfoil
Myriophyllum
*water buttercup
Ranunculus
10
fanwort
Cabomba
caraliniana Gr.
coontail
Ceratophyllum
-------
An Artificial Key to Some Common Aquatic Plants
11. Submerged leaves long 12
and ribbon-like, at
least 1/10 inch wide.
11. Submerged leaves not 22
ribbon-like; often
thread-like but if wider
than 1/10 inch, less
than 1 inch long.
12. Leaves scattered along 13
the stem.
12. Leaves all borne from
one point.
13. Leaves with mid-ribs *
evident when held against
bright light, many
species with great
diversity in leaf forms.
13. Leaves without mid-ribs
evident when held against
bright light.
14. Plants with both
floating and submerged
leaves, the floating
leaves with expanded
blades and differing
from those submerged.
14. Plants with all leaves 18
alike and submerged.
21
pondweed
Potamogeton
14
15. Floating leaves, heart-
shaped at the base, 1 to
4 inches long, waxy in
appearance.
15. Floating leaves rounded
at the base.
16. Floating leaves with
30 to 50 nerves,
submerged leaves
about three times
as long as broad.
16. Floating leaves with
less than 30 nerves.
water star grass
Heteranthera
15
17. Upper submerged leaves
with long stalks.
17. Submerged leaves not
as above but with an
abrupt awl-shaped
tip.
18. Margins of the thin
leaves crimped and
toothed, the marginal
serrations visible to
the naked eye.
18. Margins of leaves
not visibly toothed.
19. Leaves minutely toothed
on the margins, visible
when magnified, leaves
extending stiffly in
opposite directions so
that whole plant appears
flat; only midvein
prominent.
19. Not as above
floating pondweed
Potamogeton
natans L.
16
large leaf pondweed
Potamogeton
amplifolius
Tuckerman
17
20. Stems much flattened
and winged, about as
wide as the leaves;
leaves 1/12 to 1/5
inch wide.
20. Leaves threadlike, *
long, rounded, and
slender, rarely
exceeding 1/10 inch
wide, oriented into
a lax, diffuse,
branched spray. The
"bunched" appearance
of the threadlike
rounded leaves as they
float in the water
readily distinguished
sago pondweed from
others of group.
American
pondweed
Potamogeton
nodosus
Poiret
pondweed
Potamogeton
angustifolius
Berchtold
* curlyleaf
pondweed
Potamogeton
crispus L.
19
Robbins
pondweed
Potamogeton
robbinsii
Oakes
20
flatstem
pondweed
Potamogeton
zosteriformis
Fernald
sago pondweed
Potamogeton
pectinatus L.
-------
An Artificial Key to Some Common Aquatic Plants
21. Leaves very long and
ribbonlike; when
examined with hand
lens, showing a central
dense zone and a
peripheral less dense
zone; flowers borne on
a long stem that forms
a spiral after fertili-
zation.
21. Leaf, when examined
with hand lens not
showing zones as above.
22. Leaves whorled, 3-8
in upper whorls.
22. Leaves alternate or
opposite.
23. Leaves alternate, leaf-
base apparently
inflated.
23. Leaves opposite.
24. All leaves long
and narrow.
24. Upper leaves shorter
and broader.
* Vallisneria
waterplantains,
Alismataceae
Elodea, Egeria,
Hydrilla
23
widgeongrass,
Ruppia maritima L.
24
25
waterstarwort,
Callitriche
25. Leaves dilated at base. * naiad, Najas
25. Leaves with narrow
bases.
horned pondweed,
Zannichiellia
palustris L.
2. Base of stem tri-
angular in cross
section, the three
angles in some cases
so rounded as to make
the stem appear al-
most round.
2. Base of stem not
triangular.
3. Three cornered seeds,
usually straw colored,
enclosed within a loose
elongated sac, a low-
growing grasslike plant.
3. Seeds not enclosed in a
loose elongated sac.
4. A single flower or
seed-bearing struture
on the tip of the stem.
sedge, Carex
spikerush,
Eleocharis
Stem with one or more ^bulrush,
leaves extending beyond Scirpus
the spike or seed-
bearing structure.
(The hardstem bulrush
has long, hard, slender,
dark olive-green stems,
1/8 to 3/8 inch at the
base, extending 3 to 5
feet above the water
surface, the softstream
bulrush has soft stems
of light green color,
3/10 to 1 inch thick
at the base.)
C PLANTS ERECT AND EMERSED. ROOTED
TO THE SUBSTRATUM AND EXTENDING
UPWARD OUT OF THE WATER.
1. Leaves more than 10 2
times as long as broad.
1. Leaves less than 10 12
times as long as broad.
5. Leaf with a collarlike
appendage, membranous
or composed of hairs at
the junction of the leaf
blade and that part of the
leaf that is wrapped
around the stem.
5. Leaf without collarlike
appendage mentioned
above.
-------
An Artificial Key to Some Common Aquatic Plants
6. Seed or flower-bearing Cutgrass, 10.
structure composed of Leersia
scales with fringed
margins and over-
lapping in a single row.
6. Flower-bearing struc- 7
ture not as above.
Flowers in spherical
heads, seeds larger,
up to size of corn,
kernel, leaves
shallowly and
broadly triangular
in cross-section.
burreed,
Sparganium
11. Plants aromatic when sweetflag
wildrice,
Zizania
aquatica L.
13.
7. Flowering heads composed common reed,
of small seeds with long Phragmites
silky hairs, appearing as a 11.
silky mass. The root-
stocks are stout, making
it a difficult plant to pull
up. Plants are 6 to 12 feet
taU.
7. Flowering heads not
appearing as a silky
mass.
8. Flowering part of
plant much branched,
but not as closely
packed as in Phragmites.
Seeds much larger,
about 3/4 inch long.
Plants with short
roots and easily pulled
up. 13,
8. Spikelets, 6 mm long *cordgrass,
or more appressed Spartina
along one side of
rachis, grass growing
in clumps or solid
stands.
9. None of the veins more 10
prominent than others.
9. Midvein more prom- 11
inent than others.
crushed.
Plants not aromatic
when crushed,
flowers large and
showy.
12. Leaves arising at
intervals along the
stem.
12. Leaves arismg at
base of the plant.
10. Flowers borne in closely Cattail,
packed cylindrical spikes,
seeds very small. (The
common cattail has flat
leaves about 1 inch
wide, the narrow-leaved
cattail has leaves some-
what rounded on the back
that are 1/8 to 7/8 inch
wide.)
Tj^ha
A corus
calamus L.
yellow iris
Iris
pseudacorus L.
13
Plants with jointed
stems, swollen at the
joints, or with creeping
rootstocks, stems with
alternate, simple
leaves.
Stems prostrate or
creeping, branched,
and often jointed and
rooted at the joints;
leaves opposite,
spreading plant,
often forming floating
mats over extensive
water areas crowding
out other plants,
broken off branch frag-
ments root readily, and
stems may elongate as
much as 200 inches in
one season.
14
smartweed,
Polygonum
alligatorweed,
A lternanthera
philoxeroides
Mart.
15
14. Fleshy or tuber-
bearing rootstocks
and rosettes of sheathing
basal leaves.
14. Not as above, floating 16
plants.
-------
An Artificial Key to Some Common Aquatic Plants
15. None of the veins more ^arrowhead
prominent than others. Sagittaria
15. Midvein and those arrow arum
descending into the Peltandra
lobes more prominent Virginia L.
than others.
16. Plants floating with waterhyacinth
fibrous, branched roots Eichhornia
and rosettes of stalked crassipes
leaves, the leaf stalks Mart,
often inflated and
bladder-like.
5. Leaf tip 3-pointed.
shoalgrass,
Diplanthera
wrightii
water-
chestnut,
Trapa natans L.
16. Plants with floating
rosettes of stalked
leaves, commonly
several rosettes
produced on branches
of the same plant at
the end of flexible,
cardlike, sparsely-
branched submerged
stems; plant thrives
at depths of 2 to 5
feet and favors muddy
bottoms with high
organic content, leaf
stalks inflated, but
not as conspicuously
as in waterhyacinth.
D SUBMERSED "SEA GRASSES"
1. Leaves round in cross manateegrass,
section. Cymodocea
manatorum
2. Leaves flat. 3
3. Leaves widest at or Halophila
above the middle.
3: Leaves uniform width. 4
4. Leaf tip blunt.
turtlegrass,
Thalassia
testudinum
4. Leaf tip pointed. 5
5. Leaf tip single.
eelgrass,
Zostera marina
ACKNOWLEDGMENT:
This outline was adapted from Keys of
Mackenthun and Ingram (1967); Robertson
and Eyles (1963); and Humm (1956). The
figures are from Eyles and Robertson and
Hotchkiss (1967).
REFERENCES
1 Blackburn, R.D., Weldon, L.W., Yeo,
R.R., and Taylor, T.M.
Identification and Distribution of
Certain Similar-Appearing Submersed
Aquatic Weeds m Florida. Hyacinth
Control Jour.8:17-21. 1969.
2 Eyles, Don E. and Robertson, J. L.
Guide and Key to the Aquatic Plants
of the Southeastern United States.
USFWS Circular 158, 151 pp. 1963.
3 Hotchkiss, Neil. Underwater and
Floating-Leaved Plants of the United
States and Canada. USDI. Fish and
Wildlife Service Resource Publication
Number 44. 124 pp. 1967.
4 Humm, H.J. Sea Grasses of the
Northern Gulf Coast, Bull. Mar. Sci.
Gulf and Carib. 6 (4):305-308. 1956.
5 Mackenthun, Kenneth M. and Ingram,
William M. Biological Associated
Problems in Freshwater Environments.
FWPCA. 287 pp. 1967.
6 Ogden, E.C. Key to the North American
Species of Potamogeton. Circ. N.Y.
State Museum. 31:1-11. 1953.
7 Prescott, G.W. How to Know the Aquatic
Plants. Wm. C. Brown. 1969.
8 Sculthorpe, C. D. The Biology of Aquatic
Vascular Plants. St. Martin's Press.
New York. 610 pp. 1967.
-------
An Artificial Key to Some Common Aquatic Plants
9 Ward, H.B. and Whipple, G.C.
(Edited by W, T. Edmondson) 1959,
Fresh Water Biology, John Wiley and
Sons, New York, 1248 pp. (includes
chapters Aquatic Bryophyta by Conrad
and Vascular Plants by Muensher.
10 Weldon, Lyle W. Common Aquatic
Weeds. USDA. Agricultural
Handbook 352, 43 pp. 1969.
This outline was prepared by R.M. Sinclair,
Aquatic Biologist, National Training Center,
Water Programs Operations, EPA, Cincinnati,
OH 45268.
-------
An Artificial Key to Some Common Aquatic Plants
Group A
FLOATING PLANTS
Riccia
Azolla
Ricciocarpus
Salvinia
Ceratopteris
Limnobium
, . Pistia
Eichornia
C? &
Wolffia
q Lemna triscula
y c
Lemna minor Wolffiella
Spirodela
Elodea
-------
An Artificial Key to Some Common Aquatic Plants
Group A
FLOATING PLANTS (Cont.)
Nelumbo
Nymphaea
seedling, life-size
-------
An Artificial Key to Some Common Aquatic Plants
Group B
SUBMERSED
Podostemum
-------
An Artificial Key to Some Common Aquatic Plants
Group B
SUBMERSED (Cont.)
Chara
Myriophyllum
Cabomba
Ranunculus
-------
An Artificial Key to Some Common Aquatic Plants
Group B
SUBMERSED (Cont. )
Tuber
stem
detail
Ruppia
habit
stem
detail
habit
P. pectinatus
habit
POTAMOGETON
stem
detail
N. flexilis
NAJAS
P. crispus
P. natans
-------
An Artificial Key to Some Common Aquatic Plants
Group C
EMERSED
Sparganium
Zizania
stem detail
Cyperus
Spartina
Zizamopsis
Leersia
%
habit
stem
detail
root
detail
stem
detail
habit
Eleocharis
Scirpus
stem
-------
An Artificial Key to Some Common Aquatic Plants
Group D
SEA GRASSES
Diplanthera
Halophila
Phyllospadix
-------
An Artificial Key to Some Common Aquatic Plants
AQUATIC MACROPHYTES - ALGAE AND BRYOPHYTES
Chara
Division ALGAE
Nitella
"Liverworts"
Class Hepaticae
Riccia
Porella
'
-------
An Artificial Key to Some Common Aquatic Plants
AQUATIC VASCULAR PLANTS - TRACHEOPHYTA
Class
Equisetinae
Equisetum
Class Felicinae
F
Ceratopteris
Marsilea
A zolla
Nymphoides
Class Angiospermae
Myriophyllum
DICOTYLEDONS
Podostemum
I
Thalassia
Potamogeton
MONOCOTYLEDONS
-------
SECTION C
ANIMALS: CONSUMERS
Animals are consumers; they exist by eating either plants or other animals.
The consumer's carbon comes directly or indirectly from the carbon in a
plant product; the consumer's utilization of energy comes from the oxidation
of some of the plant material back to carbon dioxide and water.
Included in this section are surveys of all major groups of aquatic animals
together with a key to selected groups. Ecologically they may be placed tn
four groups: zooplankton; benthos, nekton; and periphyton, in part.
Contents of Section C
Key to Selected Groups of Freshwater Animals
Outline No.
14
Biota of Wastewater Treatment Plants
(Microscopic Invertebrates)
15
Biology of Zooplankton Communities
16
Macro Invertebrates
17
Aquatic Insects
18
Freshwater Crustacea
19
Freshwater Mollusca
20
Fishes
21
Classification of Fishes
-------
KEY TO SELECTED GROUPS OF FRESHWATER ANIMALS
The following key is intended to provide
an introduction to some of the more
common freshwater animals Technical
language is kept to a minimum
In using this key, start with the first
couplet (la, lb), and select the alternative
that seems most reasonable. If you
selected "la" you have identified the
animal as a member of the group, Phylum
PROTOZOA If you selected "lb" , proceed
to the couplet indicated Continue this
process until the selected statement is
terminated with the name of a group
If you wish more information about the
group, consult references (See reference
list )
BI. AQ. 21b. 5.71
-------
Key to Selected Groups of Freshwater Animals
la The body of the orgams.n comprising
a single microscopic independent
cell, or many similar and indepen-
dently functioning cells associated
in a colony with little or no differ-
ence between the cells, 1 e , with-
out forming tissues, or body com-
prised of masses of multinucleate
protoplasm Mostly microscopic,
single celled animals.
Phylum PROTOZOA
lb The body of the organism com- 2
prised of many cells of different
kinds, i.e., forming tissues.
May be microscopic or macro-
scopic.
2a Body or colony usually forming 3
irregular masses or layers some-
times cylindrical, goblet shaped,
vase shaped, or tree like. Size
range from barely visible to
large.
2b Body or colony shows some type 4
of definite symmetry.
3a Colony surface rough or bristly
in appearance under microscope
or hand lens. Grey, green, or
brown. Sponges.
Phylum PORIFERA (Fig. 1)
3b Colony surface relatively smooth.
General texture of mass gelatinous,
transparent. Clumps of minute
individual organisms variously
distributed. Moss animals,
bryozoans.
Phylum BRYOZOA (Fig. 2)
4a Microscopic. Action of two
ciliated (fringed) lobes at an-
terior (front) end in life often
gives appearance of wheels.
Body often segmented, accordian-
like. Free swimming or attached.
Rotifers or wheel animalcules.
Phylum TROCHELMINTHES
(Rotifera) (Fig. 3)
4b Larger, wormlike, or having 5
strong skeleton or shell.
5a Skeleton or shell present. Skel- 15
eton may be external or internal.
5b Body soft and/or wormlike. 6
Skin may range from soft to
parchment-like.
6a Three or more pairs of well 19
formed jointed legs present.
Phylum ARTHROPODA (Fig. 4)
6b Legs or appendages, if present, 7
limited to pairs of bumps or hooks.
Lobes or tenacles, if present,
soft and fleshy, not jointed.
7a Body strongly depressed or 8
flattened in cross section.
7b Body oval, round, or shaped like 10
an inverted "U" in cross section.
8a Parasitic inside bodies of higher
animals. Extremely long and flat,
divided into sections like a Roman
girdle. Life history may involve
an intermediate host. Tape worms.
Class CESTODA (Fig. 5)
8b Body a single unit. Mouth and 9
digestive system present, but no
anus.
9a External or internal parasite of
higher animals. Sucking discs
present for attachment. Life his-
tory may involve two or more in-
termediate hosts or stages. Flukes.
Class TREMATODA
9b Free living. Entire body covered
with locomotive cilia. Eye areas
in head often appear "crossed".
Free living flatworms.
Class TURBELLARIA (Fig. 6)
10a Long, slender, with snake-like
motion in life. Covered with glis-
tening cuticle. Parasitic or free-
living. Microscopic to six feet in
length. Round worms.
Phylum NEMATHELMINTHES
(Fig. 7)
10b Divided into sections or segments 11
-------
Key to Selected Groups of Freshwater Animals
10c Unsegmented, head blunt, one 18
or two retractile tentacles.
Flat pointed, tail.
11a Head a more or less well-formed,
hard, capsule with jaws, eyes,
and antennae.
Class INSECTA order DIPTERA
(Figs. 8A, 8C)
lib Head structure soft, except
jaws (if present). Fig. 8E.)
12
12a Head conical or rounded, lateral 13
appendages not conspicuous or
numerous.
12b Head somewhat broad and blunt. 14
Retractile jaws usually present.
Soft fleshy lobes or tentacles,
often somewhat flattened, may be
present in the head region. Tail
usually narrow. Lateral lobes
or fleshy appendages on each
segment unless there is a large
sucker disc at rear end.
Phylum ANNELIDA (Fig. 9)
13a Minute dark colored retractile
jaws present, body tapering
somewhat at both ends, pairs or
rings of bumps or "legs" often
present, even near tail.
Class INSECTA Order DIPTERA
(Fig. 8)
13b No jaws, sides of body generally 14
parallel except at ends. Thicken-
ed area or ring usually present
if not all the way back on body.
Clumps of minute bristles on most
segments. Earthworms, sludge-
worms.
Order OLIGOCHAETA
14a Segments with bristles and/or fleshy
lobes or other extensions. Tube
builders, borers, or burrowers.
Often reddish or greenish in
color. Brackish or fresh water.
Nereid worms.
Order POLYCHAETA (Fig. 9A)
14b Sucker disc at each end, the large
one posterior. External blood-
sucking parasites on higher animals,
often found unattached to host.
Leaches.
Class HIRUDINEA (Fig. 9B)
15a Skeleton internal, of true bone. 40
(Vertebrates)
15b Body covered with an external 16
skeleton or shell.
(Figs. 10, 13, 17, 18, 24,
25, 28)
16a External skeleton jointed, shell 19
covers legs and other appendages,
often leathery in nature.
Phylum ARTHROPODA
16b External shell entire, not jointed, 17
unless composed of two clam-
like halves.
(Figs. 10, 11, 12)
17a Half inch or less in length. Two
leathery, clam-like shells. Soft
parts inside include delicate,
jointed appendages. Phyllopods
or branchiopods.
Class CRUSTACEA, Subclasses
BRANCHIOPODA (Fig. 12)
and OSTRACODA (Fig. 11)
17b Soft parts covered with thin 18
skin, mucous produced, no jointed legs.
Phylum MOLLUSCA
18a Shell single, may be a spiral cone.
Snails.
Class GASTROPODA (Fig. 13)
18b Shell double, two halves, hinged
at one point. Mussels, clams.
Class BIVALVIA (Fig. 10)
19a Three pairs of regular walking 29
legs, or their rudiments. Wings
present m all adults and rudiments
in some larvae.
Class INSECTA (Figs. 22, 24D,
25, 26, 28, 29)
19b More than three pairs of legs 20
apparently present.
20a Body elongated, head broad and flat
-------
Key to Selected Groups of Freshwater Animals
with strong jaws. Appendages follow-
ing first three pairs of legs are round-
ded tapering filaments. Up to 3
inches long. Dobson fly and fish fly
larvae.
Class INSECTA Order
MEGALOPTERA (Fig. 14)
20b Four or more pairs of legs. 21
21a Four pairs of legs. Body rounded ,
bulbous, head minute. Often
brown or red. Water mites.
Phylum ARTHROPODA, Class
ARACHNIDA, Order ACARI
(Fig. 15)
21b Five or more pairs of walking 22
or swimming legs; gills, two
pairs of antennae. Crustaceans.
Phylum ARTHROPODA,
Class CRUSTACEA
22a Ten or more pairs of flattened,
leaflike swimming and respiratory
appendages. Many species swim
constantly in life, some swim
upside down. Fairy shrimps,
phyllopods, or branchipods.
Subclass BR ANCHIOPODA
(Fig. 16)
22b Less than ten pairs of swimming 23
or respiratory appendages.
24
23a Body and legs inclosed in bi-
valved (2 halves) shell which may
or may not completely hide them.
23b Body and legs not enclosed in 26
bivalve shell. May be large or
minute.
(Figs. 17, 18, 19)
24a One pair of branched antennae
enlarged for locomotion, extend
outside of shell (carapace).
Single eye usually visible.
"Water fleas"
Subclass CLADOCERA (Fig. 12)
24b Locomotion accomplished by 25
body legs, not by antennae.
25a Appendages leaflike, flattened,
more than ten pairs.
Subclass BRANCHIOPODA
(See 22 a)
25b Animal less than 3 mm, in length.
Appendages more or less slender
and jointed, often used for walking.
Shells opaque. Ostracods.
(Fig. 11) Subclass OSTRACODA
26a Body a series of six or more 27
similar segments, differing
mainly in size.
26b Front part of body enlarged into 28
a somewhat separate body unit
(cephalothorax) often covered
with a single piece of shell (cara-
pace). Back part (abdomen) may be
relatively small, even folded
underneath front part. (Fig. 19b)
27a Body compressed laterally, i.e.,
organism is tall and thin. Scuds,
amphipods.
Subclass AMPHIPODA (Fig. 17)
27b Body compressed dorsoventrally,
i.e., organism low and broad.
Flat gills contained in chamber
beneath tail. Sowbugs.
Subclass ISOPODA (Fig. 18)
28a Abdomen extending straight out
behind, ending in two small pro-
jections. One or two large masses of
eggs are often attached to female.
Locomotion by means of two enlarged,
unbranched antennae, the only large
appendages on the body. Copepods.
Subclass COPEPODA (Fig, 19)
28b Abdomen extending out behind ending
in an expanded "flipper" or swim-
ming paddle. Crayfish or craw fish.
Eyes on movable stalks. Size range
usually from one to six inches.
Subclass DECAPODA
29a Two pairs of functional wings, 39
one pair may be more or less har-
dened as protection for the other
pair. Adult insects which normally
live on or in the water. (Figs. 25, 28)
-------
Key to Selected Groups of Freshwater Animals
29b No functional wings, though 30
pads in which wings are develop-
ing may be visible. Some may
resemble adult insects very
closely, others may differ ex-
tremely from adults.
30a External pads or cases in which 35
wings develop clearly visible.(Figs.
24, 26, 27)
30b More or less wormlike, or at 31
least no external evidence of
wing development.
31a No jointed legs present. Other
structures such as hooks, sucker
discs, breathing tubes may be
present. Larvae of flies,
midges, etc.
Order DIPTERA (Fig. 8)
31b Three pairs of jointed thoracic 32
legs, head capsule well formed.
32a Minute (2-4mm) living on the
water surface film. Tail a
strong organ that can be hooked
into a "catch" beneath the
thorax. When released animal
jumps into the air. No wings
are ever grown. Adult spring-
tails.
Order COLLEMBOLA (Fig. 20)
32b Larger (usually over 5 mm) 33
wormlike, living beneath the
surface.
33a Live in cases or webs in water.
Cases or webs have a silk
foundation to which tiny sticks,
stones, and/or bits of debris
are attached. Abdominal segments
often with minute gill filaments.
Generally cylindric in shape.
Caddisfly larvae.
Order TRICHOPTERA (Fig. 21)
33b Free living, build no cases. 34
34a Somewhat flattened in cross
section and massive in appear-
ance. Each abdominal segment
with rather stout, tapering, lateral
filaments about as long as body
is wide. Alderflies, fishflies, and
dobsonflies.
Order MEGALOPTERA (Fig. 22, 14)
34b Generally rounded in cross section.
Lateral filaments if present tend
to be long and thin. A few forms
extremely flattened, like a suction
cup. Beetle larvae.
Order COLEOPTERA (Fig. 23)
35a Two or three filaments or other 37
structures extending out from
end of abdomen.
35b Abdomen ending abruptly, unless 36
terminal segment itself is extended
as single structure.(Figs. 24A, 24C)
36a Mouth parts adopted for chewing.
Front of face covered by extensible
folded mouthparts often called a
"mask". Head broad, eyes widely
spaced. Nymphs of dragonflies
or darning needles.
Order ODONATA (Figs.24A, 24C, 24E)
36b Mouthparts for piercing and sucking.
Legs often adapted for water lo-
comotion. Body forms various.
Water bugs, water scorpions, water
boatmen, backswimmers, electric
light bugs, water striders, water
measurers, etc.
Order HEMIPTERA (Fig. 25)
37a Tail extensions (caudal filaments)
two. Stonefly larvae.
Order PLECOPTERA (Fig. 26)
37b Tail extensions three, at times 38
greatly reduced in size.
38a Tail extensions long and slender.
Rows of hairs may give extensions
a feather-like appearance.
Mayfly larvae.
Order EPHEMEROPTERA
(Fig. 27)
38b Tail extensions flat, elongated
plates. Head broad with widely
spaced eyes, abdomen relatively
long and slender. Damselfly
nymths.
Order ODONATA (Fig. 24D^
-------
Key to Selected Groups of Freshwater Animals
39a External wings or wing covers
form a hard protective dome
over the inner wings folded
beneath, and over the abdomen
Beetles.
Order COLEOPTERA
(Fig 28)
39b External wings leathery at base.
Membranaceous at tip Wings
sometimes very short, Mouth-
parts for piercing and sucking
Body form various True bugs.
Order HEMIPTERA (Fig 25)
40a Appendage present in pairs
(fins, legs, wings)
42
42a Paired appendages are legs 43
42b Paired appendages are fins,
S'lls covered by a flap
perculum) True fishes
Class PISCES
43a Digits with claws, nails, or hoofs 44
43b Skin naked. No claws or digits
Frogs, toads, and salamanders
Class AMPHIBIA
44a Warm blooded 45
40b No paired appendages,
a round suction disc
Mouth 41 44b Cold blooded Body covered
with horny scales or plates
Class REPTILIA
41a Body long and slender Several
holes along side of head
Lampreys.
Sub Phylum VERTEBRATA,
Class CYCLOSTOMATA
41b Body plump, oval. Tail extending
out abruptly. Larvae of frogs and
toads Legs appear one at a time
during metamorphosis to adult
form. Tadpoles.
Class AMPHIBIA
45a Body covered with feathers
Birds
Class AVES
45b Body covered with hair
Mammals
Class MAMMALIA
-------
Key to Selected Groups of Freshwater Animals
REFERENCES - Invertebrates
1 Eddy, Samuel and Hodson, A.C.
Taxonomic Keys to the Common
Animals of the North Central States.
Burgess Pub. Company, Minneapolis.
162 p. 1961.
2 Edmondson, W. T.
-------
Key to Selected Groups of Freshwater Animals
1. Spongilla spicules
Up to . 2 mm. long.
3C. Rotifer, Philodina
Up to . 4 mm.
3A. Rotifer, Polyarthra
Up to .3 mm.
3B. Rotifer. Keratella
Up to . 3 mm.
2B. Bryozoal mass. Up to
several feet diam.
Bryozoa, Plumatella. Individuals up
to 2 mm. Intertwined masses maybe
very extensive.
4C. Jointed leg
Ostracod
4B. Jointed leg
Crayfish
4A. Jointed leg
Caddisfly
5. Tapeworm head,
Taenia. Up to
25 yds. long
6B. Turbellaria, Dugesia
Up to 1. 6 cm.
7. Nematodes. Free living
forms commonly up to
1 mm., occasionally
more.
-------
Key to Selected Groups of Freshwater Animals
9
8B Diptera, Mosquito
pupa Up to 5mm,
8A. Dipt(.rd, Mosquito larvae
Up to 15 mm. long.
8C. Diptera, chironomid 8E Dipte";
, pupa Up to 2 5 cm
larvae. Up to 2 cm.
9D. Diptera, Rattailed maggot
Up to 25 mm. without tube.
Annelid
gmented
30B. Alasmidonta, end view.
I OA. Pelecyopod, Alasraidonta
Side view, up to 18 cm.long.
9B. Annelid, leech up to 20 cm.
a7u 12A. Branchiopod,
Daphnia. Up
to 4mm.
UA. Ostraeod, Cypericus
Side view, up to 7 mm.
1 IB. Cypericus, end view,
12B. Branchiopod,
Bosmina. Up
to 2mm.
-------
Key to Selected Groups of Freshwater Animals
13. Gastropod, Campeloma
Up to 3 inches.
15. Water mite,
up to 3 mm.
14. Megaloptera, Sialis
Alderfly larvae
Up to 25 mm.
16. Fairy Shrimp, Eubranchipus
Up to 5 cm.
17. Amphipod, Pontoporeia
Up to 25 mm.
18. Isopod, Asellus
Up to 25 mm.
20. Collembola, Podura
Up to 2 mm. long
10
19A. Calanoid copepod, ,
Female 19B. Cyclopoid copepodr
Up to 3 mm. Female
-------
Key to Selected Groups of Freshwater Animals
21A.
1 IL
21B.
21C,
21D. 21E.
21. Trichoptera, larval cases,
mostly 1-2 cm.
/ /fw!
22. Megaloptera,• alderfly
Up to 2 cm.
23A„ Beetle larvae, 23B. Beetle larvae, 24A. Odonata, dragonfly
Dytisidae, Hydrophilidae nymph up to 3 or
Usually about 2 cm. ^sually about 4 cm
24B. Odonata, tail
of damselfly
nymph
(side view)
Suborder
Zygoptera
(24B, D)
\ 24D. Odonata, damselfly
nymph (top view)
24E, Odonata, front view
/ of dragonfly nymph
showing "mask"
partially extended
Suborder
Anisoptera
(24A, E, C)
24C. Odonata, tail of
dragonfly nymph
(top view)
-------
Key to Selected Groups of Freshwater Animals
25A. Hemiptera,
Water Boatman
About 1 cm.
25B. Hemiptera,
Water Scorpion
About 4 cm.
26. Plecoptera,
St one fly nymph
Up to 5cm.
27.Ephemeroptera.
Mayfly nymph
Up to 3cm.
2BA. Coleoptera,
Water scavenger
beetle. Up to 4 cm.
28B.
Coleoptera,
Dytiscid beetle
Usually up to 4
cm,
29A. Diptera, Crane
fly. Up to 2i cm.
29B. Diptera, Mosquito
Up to 20 mm.
-------
BIOTA OF WASTEWATER TREATMENT PLANTS
(MICROSCOPIC INVERTEBRATES)
I GENERAL CONSIDERATIONS
A Community rather than individual as a
unit for study of the process, quantitative
relationship among different populations -
"population dynamics".
B Sequential transformation of organic
matter through the microbial life - a
transference of materials and energy
between microbial populations led to the
development of functional synecology or
productive ecology
C Microbes considered here include bacteria,
protozoa, and microscopic metazoa, algae
and fungi are important groups included
elsewhere in more detail.
D All microbial groups originate from
a) the waste itself, b) washing waters,
c) soil, d) dust from air, and e) incidental
sources, only those members that can
survive and establish themselves in the
community are important, some are
transient.
E Some variations in composition of the
microbial community in domestic sewage
treatment due to climatic and other
ecological factors, industrial wastes with
specific waste matter may call for develop-
ment of more restricted microbial com-
munity for degradation.
«
F Most active microbial groups are True
bacteria, filamentous bacteria, fungi,
protozoa, nematodes, rotifers, oligochaetes,
and water-mites.
II BACTERIA
A No ideal method for studying distribution
and ecology of bacteria in waste-treatment.
Total bacterial counts made on nutrient
agar or gelatine reflect only a portion of
the bacterial flora present.
B Pseudomonads are probably the most
versatile in their ability to attack a great
variety of organic compounds, including
petroleum products, phenolics, cyanides.
Others, such as Achromobacter, Alca-
ligenes, Chromobacterium, Flavobacterium,
Aerobacter, and Micrococcus, are also
important genera Actinomyces are
prominent in wastes rich in cellulose and
Bacillus organisms are starch attackers.
Sulfur and iron bacteria are predominant
in wastes rich in respective compounds
C Actinomyces, Bacillus spp , Aerobacter
spp. , and nitrogen-fixation bacteria are
primarily soil dwellers and are almost
always present in any type of wastes in
small numbers.
D Parasitic and pathogenic bacteria, if
present, are transient.
E In extended aeration process with high
dissolved oxygen, predominant species
are limited to pseudomonads, Zoogloea
ramigera, and Sphaerotilus
III PROTOZOA
A Classification
1 Single-cell animals in the phylum
Protozoa in the animal kingdom.
or
2 A separate kingdom. Protista, to
include protozoa, algae, fungi, and
bacteria.
a Mastigophora (flagellates) - only
the subclass Zoomastigina (non-
pigmented) included, four orders:
SE BI. 4e. 11. 72
-------
Biota of Wastewater Treatment Plants
1) Rhizomastigina - amoeba-
flagellates with 1 or more
flagella, examples Mastigamoeba
Actinomonas, Rhizomastix
2) Protomonadina - with 1 or 2
flagella, comprising most of the
free-livmg forms, examples-
Peranema, Bodo, Monas,
Pleuromonas
3) Polymastigina - with 3-8 flagella,
mostly parasitic in gut of man
and animals
4) Hypermastigina - with numerous
flagella, all parasitic in insect
intestine
b Ciliophora or Infusoria (ciliates) -
largest class of protozoa, no pig-
mented members, most important
group of protozoa in waste treatment,
2 subclasses.
1) Ciliata - cilia present during the
the entire trophic life, comprising
most of the common ciliates,
examples- Paramecium, Colpi-
dium, Colpoda, Euplotes,
Stylonychia, Vorticella, Oper-
culana, Epistylis, Carchesium
2) Suctona - cilia present while
young and tentacles during
trophic life
c Sarcodina (amoebae) - pseudopodia
(false feet) for locomotion and food-
capturing, cell without cell-wall,
some with test or shell, 2 subclasses
1) Rhizopoda - pseudopodia without
axial filaments, 5 orders-
a) Proteomyxa - with radiating
pseudopodia, no test or shell
b) Mycetazoa (slime-molds)
forming plasmodium, re-
sembling fungi in sporangium
formation.
c) Amoebina - true amoeba,
pseudopodia in the form of
lobopodia, no test or shell,
cyst formation frequent, a few
capable of flagellate trans-
formation, examples: Naegleria,
Amoeba, Hartmannella,
Endamoeba
d) Testacea - amoeba with single
test or shell, examples
Arcella, Difflugia
e) Forammifera - large amoeba
with calcareous shell, all
marine forms
2) Actinopoda - with spinous
pseudopodia, 2 orders
a) Heliozoa - without central
capsule, usually spherical in
form with many radiating
axopodia, examples Actino-
sphaerium, Actinophrys
b) Radiolaria - pelagic in various
oceans
d Sporozoa - no organ of locomotion,
all parasitic (Plasmodium, Coccidia)
B General Morphology
1 Zoomastigina:
With the exception of Rhizomastigina
which is amoeboid, the body has defi-
nite shape (oval, leaf-like, pear-like,
etc.); most free-living forms with l->2
flagella, some with 3 or more flagella,
few forming colonies, cytostome present
in many for feeding on bacteria, rela-
tively small size (15-40 \i)
2 Ciliophora
Most highly developed protozoa, with
few exceptions, a macro- and a micro-
nucleus, adoral zone, mouth, oral
groove, usually present in swimming
and crawling forms, stalked form with
-------
Biota of Wastewater Treatment Plants
conspicuous ciliation of a disc-like
anterior region and little or no body
cilia, cyst formed in most species
3 Sarcodina
Cytoplasmic membrane but not cell-
wall, cytoplasm with distinct ectoplasm
and endoplasm in many common spp ,
nucleus with large nucleolus in most
of the free-living forms, some with
the body enclosed in a test or shell
and moving by protruding pseudopodia
outside of the enclosure through an
opening, few capable of temporary
transformation into flagellate, fresh-
water actinopods usually spherical
with many radiating axopodia, some
Testacea spp. containing symbiotic
algae - mistaken for pigmented amoebae,
cysts with single or double wall and
1-2 nuclei, parasitic amoebae forming
cysts with 4 or more nuclei
C General Physiology
1 Zoomastigina
I
Free-living forms normally holozoic,
food supply mostly bacteria, relatively
aerobic, therefore, among the first to
disappear in anaerobic conditions, re-
production by simple fission and
occasionally by budding.
2 Ciliophora
Holozoic, true ciliates concentrating
food particles, 1. e , bacteria, by
ciliary movement around the mouth-
part, suctoria sucking through tentacles,
bacteria, small algae and protozoa
constituting main food under normal
conditions, not as aerobic as flagellates -
a few surviving under highly anaerobic
conditions, such as Metopus, repro-
ducing by simple fission, conjugation,
or encystation.
3 Sarcodina
Mostly holozoic, feeding through engulf-
ing by pseudopodia, food supply of small
amoebae mostly bacteria, large
amoebae engulfing larger organisms,
shelled amoebae, i.e , Arcella. feeding
on a variety of organisms or saprozoic,
reproduction by simple fission and
encystation.
IV NEMATODA
A Classification
1 All in the phylum Nemata (nonsegmented
round worms), 2 subphyla:
Secernentea (phasmids) 6 orders
Tylenchida (spear in mouth), Rhabditida
(rhabditoid eosophagus), Strongylida
(parasitic), Ascaridida (parasitic),
Spirurida (parasitic), and Camallanida
(parasitic), with the exception of
tylenchids, all with papillae on male
tail
Adenophora (aphasmids) 5 orders-
Dorylaimida (spear in mouth), Chromo-
dorida, Monhysterida, Enoplida, and
Diocytophymatida, no papillae on male
tail, no excretary canal
2 Nematodes encountered in polluted
water and in sewage treatment mostly
belonging to order Rhabditida and few
in orders Dorylaimida and Tylenchida,
those in Rhabditida being bacteria -
feeders and those in the latter two,
feeding on algae and other zoomicrobes,
examples of rhabditids Rhabditis,
Diplogaster, Diplogasteroides, Mono-
choides, Cephalobus, Cylmdrocorpus,
Turbatrix, examples of the other two
Dorylaimus, Aphelenchoides
B General Morphology
Round, slender, nonsegmented (some with
markings on outside), most of the free-
living forms microscopic in size although
dorylaimids up to several mm in length,
sex separated but some parthenogenetic,
complete alimentray tract with elaborate
mouth parts with or without spear (or
stylet), no circulatory or respiratory
system
-------
Biota ot Wastewater Treatment Plants
C General Physiology
Most sewage treatment plant dwellers
feeding on bacteria, others preying on
protozoa, small nematodes, rotifers,
etc , clean water species vegetarians,
DO diffused through cuticle, rhabditids
tolerating lower DO than clean water spp,
reproduction - eggs - larvae 4 molts) -
adults
V ROTIFERS
A Classification.
1 Classified either as a class of the
phylum Aschelminthes (various forms
of worms) or as a separate phylum
(Rotifera), commonly called wheel
animalcules, on account of circular appearing
movement of cilia around head (corona),
corona contracted when crawling or
swimming and expanded when attached
to catch food.
2 Of the 3 classes, 2 (Seisomdea and
Bdelloidea) grouped by some authors
under Digononta (2 ovaries) and the
other being Monogononta (1 ovary),
Seisomdea containing mostly marine
forms
3 Class Bdelloidea containing 1 order
(Bdelloida) with 4 families, Philodmedae
being the most important
4 Class Monogononta comprising 3 orders-
Ploima with 14 families, Flosculariaceae
with 4 families, and Collothecaceae
with 1 family, most important genera
included in the order Ploima (1. e.,
Brachionus, Keratella, Monostyia,
Tnchocerca, Asplanchna. Polyarthra,
Synchaeta, Microcodon), common genera
under the order Flosculariaceae Floscu-
laria, Limnias, Conochilus, and Atrochus
5 Unfortunately orders and families of
rotifers based on character of corona
and trophi (chewing organ), which are
difficult to study, esp the latter, the
foot and cuticle much easier to study
B General Morphology and Physiology
1 Body weakly differentiated into head,
neck, trunk, and foot, separated by
folds, in some, these regions are
merely gradual changes in diameter
of body and without a separate neck,
segmentation external only
2 Head with corona, dosal antenna, and
ventral mouth, mastax, a chewing
organ, located in head and neck, con-
nected to mouth anteriorly by a ciliated
gullet and posteriorly to a large stomach
occupying much of the trunk.
3 Common rotifers reproducing partheno-
genetically by diploid eggs, eggs laid
in water, cemented to plants, or carried
on femals until hatching.
4 Foot, a prolongation of body, usually
with 2 toes, some with one toe, some
with one toe and an extra toe-like
structure (dorsal spur)
5 Some, like Philodina, concentrating
bacteria and other microbes and minute
particulate organic matter by corona,
larger microbes chewed by mastax,
some such as Monostyia feeding on
clumped matter, such as bacterial
growth, fungal masses, etc at bottom,
virus generally not ingested - apparently
undetected by cilia.
6 DO requirement somewhat similar to
protozoa, some disappearing under
reduced DO, others, like Philodina,
surviving at as little as 2 ppm DO.
VI SANITARY SIGNIFICANCE
A Pollution tolerant and pollution nontolerant
species - hard to differentiate - requiring
specialist training in protozoa, nematodes,
and rotifers.
B Significant quantitative difference in clean
and polluted waters - clean waters contain-
ing large variety of genera and species
but quite low in densities.
-------
Biota of Wastewater Treatment Plants
C Aerobic sewage treatment processes
(trickling filters and activated sludge
processes, even primary settling) ideal
breeding grounds for those that feed on
bacteria, fungi, and minute protozoa and
present in very large numbers, effluents
from such processes carrying large
numbers of these zoomicrobes, natural
waters receiving such effluents showing
significant increase in all 3 categories
D Possible Pathogen Carriers
1 Amoebae and nematodes grown on
pathogenic enteric bacteria in lab,
none alive in amoebic cysts, very few
alive in nematodes after 2 days after
ingestion, virus demonstrated in
nematodes only when very high virus
concentrations present, some free-
living amoebae parasitizing humans.
2 Swimming ciliates and some rotifers
(concentrating food by corona) ingesting
large numbers of pathogenic enertic
bacteria, but digestion rapid, no evidence
of concentrating virus, crawling ciliates
and flagellates feeding on clumped
organisms.
3 Nematodes concentrated from sewage
effluent in Cincinnati area showing
live E coli and streptococci, but no
human enteric pathogens
VII EXAMINATION OF SEWAGE TREATMENT
EFFLUENT, AND SLUDGE FOR MICROBES
the method is not applicable to sewage
treatment, sludge, or effluent
1 Waste treatment - the method bound
to be qualitative, material scraped
from stones in trickling filters or the
floe masses in activated sludge examined
in slide-coverslip preparations for
poor, moderate, or rich zoobiota,
material relatively rich in zoobiota
indicating satisfactory treatment
process, protozoa, rotifers, and
nematodes predominant, especially
protozoa, bristle worms and watermites
in smaller numbers, springtails and
insect larvae present as grazing fauna
on top of trickling filters
2 Sludge - representative samples sus-
pended in known quantities of dilution
water and thoroughly shaken, filtered
through bolting cloth or metal screen
of comparable pore size to remove
extraneous dead clumped matter,filtrate
examined in Sedgewick Rafter (SR) counting
cell for various zoormcrobes, fresh
sludge desired or samples refrigerated
3 Sewage effluent - samples "fixed" with
formalin, merthiolate, or similar
chemical not desirable for examination
for zoomicrobes, 50-200 ml filtered
through a 7 - or 14-micron membrane
and strained material washed with a
few mis of dilution and examined in
an SR cell for zoo-microbes quantita-
tively or qualitatively
A Bacteria - Not Included
B Zoomicrobes -
The 12th edition of the Standard Methods
(1965) has a part on Biologic Examination
of Water, Wastewater, Sludge, and Bottom
Materials, in which the sludge of sewage
treatment is discussed, but very briefly.
Much of the materials are concerned with
sediment at bottom of natural bodies of
water Chang described a method for
examination of water for nematodes, but
VIII USE OF ZOOMICROBES AS POLLUTION
INDEX
A Idea not new, protozoa suggested long ago,
many considered impractical because of
the need of identifying pollution-intolerant
and pollution-tolerant species - proto-
zoologist required.
B Can use them on a quantitative basis -
nematodes, rotifers, and nonpigmented
protozoa present in small numbers in
clean water Numbers greatly increased
-------
Bioia of Wastewater Treatment Plants
when polluted with effluent from aerobic
treatment plant or recovering from sewage
pollution, no significant error introduced
when clean-water members included in the
enumeration if a suitable method of com-
puting the pollution index developed
C Most practical method involves the
equation (A + B)/A = Z.P.I. , where
A = number of pigmented protozoa,
B = other zoomicrobes, in a unit volume
of sample, and Z.P.I. = zoological pollu-
tion index. For relatively clean water,
the value of Z P I. close to 1, the larger
the value above 1, the greater the pollution
by aerobic effluent, or sewage during
recovery This is based on the fact that
pigmented protozoa are members of clean
water micro-fauna (stabilization pond
excluded) a
IX CONTROL
A Chlonnation of Effluent and Settling
B Prolongation of Detention Time of Effluent
C Modification of Waste Treatment
D Elimination of Slow Sand Filters in
Nematode Control
X LIST OF COMMON ZOOLOGICAL ORGAN-
ISMS FOUND IN SEWAGE TREATMENT
PROCESS - TRICKLING FILTERS AND
ACTIVATED SLUDGE PROCESS
PROTOZOA
Sarcodina - Amoebae
Amoeba proteus, A radiosa
Hartmanella spp.
Arcella vulgaris
Naegleria gruben
Actinophrys sol
FLAGELLATA
Bodo caudatus
Pleuromonas jaculans
Oikomonas termo
Cercomonas longicauda
Peranema trichophorum
Swimming type
Ciliophora
Colpidium colpoda
Colpoda cucullus
Glaucoma
Paramecium caudaturn; P. bursaria
Stalked type
Opercularia spp. (short stalk
dichotomous)
Vorticella spp. (stalk single and
contractile)
Epistylis plicatilis (like Opercularia
more colonial)
Carchesium spp. (like Vorticella but
colonial, both have
spiral coiled stalk
Crawling type when contracted)
Euplotes spp.
Stylonychia mylitus
Urostyla spp.
Oxytricha spp.
NEMATODA
Diplogaster spp.
Monochoides spp.
Diplogasteroides spp.
Rhabditis spp.
Pelodera spp.
Aphelenchoides sp.
Dorylaimus sp.
-------
Biota of Wastewater Treatment Plants
Cylindrocorpus sp.
Cephalobus sp.
Rhabdolaimus sp.
Monhystera sp.
Trilobus sp.
ROTATORIA
Diglena
Monstyla
Polyarthra
Philodina
Keratella
Brachionus
OLIGOCHAETA (bristle worms)
Aelosoma hemprichi (Aelosomatidae)
Aulophorus vagus (Naididae)
Tubifex tubifex (Tubificidae)
Pachydnlus lineatus (Enchytraeidae)
INSECT LARVAE
Metnocnemus ssp. (midge)
Orthocladius ssp. (midge)
Psychoda spp. (filter fly)
OTHER ARTHROPODA
Hydrochna sp. (Acarina, mite)
Platysieus tenuipes (A carina, mite)
Hypogastrura (= Achorutes sub-viatica)
viaticus (Collembola, Springtail)
Folsomia sp. (Collembola, Springtail)
Tomoccrus sp. (Collembola, Springtail)
MOLLUSCA
Lymnaea ssp. (pulmonate snail)
Physa sp. (pulmonate snail)
XI POPULATION DYNAMICS AND THE FOOD
CHAIN IN AEROBIC SEWAGE TREATMENT
PROCESSES (Figure 1 and 2)
A Aerobic bio-oxidation of waste materials
comparable to a food chain through which
the dead organic matter is converted to
inorganic matter during the stabilization
process, e.g., waste organic matter—
bacterial phase —zoological phase —*-
inorganic matter.
B Systematics, physiology and biochemistry
involved in explaining the "chain reaction",
knowledge inadequate and fragmental,
ecological study limited to principles
governing the relationship of different
groups of flora and fauna with each other
and with the environment
C With adequate DO supply, bacterial popu-
lation increases rapidly in the presence
of rich organic food, flagellates and
amoebae, which feed on bacteria and
other small particulate matter in clumped
material (such as growth film and floe
masses), first show increase in population
size, as suspended bacterial population
increases to a high level, swimming
dilates, which feed actively on the
suspended bacteria, also increase, in-
creased consumption of bacteria and
reduced supply of dead organic matter
results in decline in bacterial population,
which, in turn, results in a decline in
the swimming ciliate population, the
presence of large populations of small
protozoa (ciliates, flagelLates, and
amoebae) results in an increase in
populations of rotifers, nematodes,
stalked ciliates, and crawling ciliates,
which feed on the small protozoa and
bacteria that are lodged in clumped
masses, eventually, scavengers, such
as mites, shelled amoebae, certain
nematodes, and bristle worms become
predominant, and bacteria and small
protozoa populations drop to the pre-
cycle level, rotifers that can concentrate
bacteria in suspension, (such as Rotifer
and Philodina, and nematodes), which
have long surviving time, may remain
for a long time; these zoomicrobes
appear in the effluent in proportion to
their respective population during treat-
ment - nematodes, rotifers, ciliates
predominant with small numbers of
flagellates and amoebae, bristle worms •
unpredictable, mites few
-------
Biota of Wastewater Treatment Plants
Raw Sewage
Insects
Oligochaetes
insect larvae
Suspended organic matter
(by hydrolysis)
Nematodes
& rotifers
Dissolved organic matter
Nonpigmented
protozoa
(respiration,
deamination,
decarboxylation, etc.)
Hete rotrophic
bacteria
Inorganic C, P, N,
S comp.
Fungi
Algae
Autotrophic bacteria
(Nitrification, sulfur
&. iron bacteria)
Pathogenic organisms
Food Chain in Aerobic Sewage Treatment Processes
Figure 1.
-------
Biota of Wastewater Treatment Plants
Organic Matter (dead)
Total Bacteria Population
Special Characteristics
Stalked Ciliaces
Crawling dilates
Rotifers and
Worms (nematodes and
oligochaetes)
Mites (Platysleus)
More "binding" organisms
(stalked dilates) in
activated sludge process
c
o
Oligochaetes, mites,
and spring talis -
grazing fauna on
trickling filters
a
o.
o
GO
3
Swimming
Clllates
Mites
Oligochaetes
Shelled Amoebae
Crawling Clliates
(Scavengers)
TIME
Population dynamics in Aerobic Sewage Treatment Process
Figure 2
D Since sewage effluent from aerobic treat-
ment processes are rich in nonpigmented
zoomicrobes, discharge of effluent into
natural causes great increase in their
members, unpolluted waters usually have
a much higher algae-to-nonpigmented-
zoomicrobes ratio The great increase
in the latter in water resulted from effluent
pollution is likely to change this ratio,
thus giving the basis for the Z. P. I. This
analysis is not applicable to stabilization
ponds due to the large algal population
present in their effluents
-------
Microbial Agents
Utilization
Treatment Process Pollution Indicator
Stabilization of Organic Matter
Aerobic
Anaerobic
Breakdown of Specific Industrial
Wastes
Stabilization Ponds
Aerobic and Anaerobic
Combined
Nitrification - Aerobic
Dentrification when
Anaerobic
Elimination of Nitrogen Through
N2 Formation - Special
Dentrification
Sludge Digestion
Aerobic
Anaerobic
Phosphorus Removal
Essential in Preventing
Algae Growth
(Composting) - Solid Wastes
Total Bacteria Count
Coliform Density
MF Counts
MPN Tube Tests
E. Coli Density
Tube Test
MF Counts
Streptococci
Tube Test
MF Counts
Infrared Spectrophotometer
Specificity
Time Requirement
Enteric Pathogens
Salmonella Spp.
Quantitation in
Mixed Population
Enteroviruses
Large Volumes
Required
Coli Phage
Type Specificity To Be
Considered
Fluorescent Antibody
Against E. Coli
Against Enteric Pathogens
Gas Chromatography
on Metabolic Products
Zoological Pollution Index
Figure 3
Elimination
Removal
Detention
Settling
Limited to Large Organisms
Flocculation
Alum
Iron
Lime
"Coagulant Aid"
Filtration
Adsorption
Straining
Medium
Reverse Osmosis
Metal Strainer
Mesh Size Limit
Antagonistic Agent
Pseudomonads
Bacterial Feeders
Protozoa
Nematodes
Rotifers
Flotation
Destruction
or
Disinfection
Chemical
Clorine
Bromine
Iodine
Ozone
Silver
Copper
Physical
Heat
UV
Gamma
Radiation
Wave
Motion
Electro-
Hydraulic
Treatment
Aeration Cationic
Anionic Detergent Detergent
-------
Biota of Wastewater Treatment Plants
REFERENCES 10
1 American Public Health Association,
American Water Works Association
and Water Pollution Control
Federation Standard Methods for
the Examination of Water and Waste-
water, 12th ed New York 1965 11
2 Chang, S. L , et al Survey of Free-
Living Nematodes and Amoebas in
Municipal Supplies J A W W A
52 613-618 12
3 Change, S L and Kabler, P W
Free-Living Nematodes in Aerobic
Treatment Plant Effluents
J W P C F 34 1256-1261 1963
13
4 Edmondson, W T , et al Ward Whipple's
Fresh Water Biology, 2nd ed
John Wiley & Sons, New York,
pp. 368-401 1959
5 Hawkes, H A Ecology of Activated
Sludge and Bacteria Beds (in Waste 14
Treatment) Pergamon Press,
pp 52-98 1960
6 Hawkes, H A The Ecology of Waste-
water Treatment, Pergamon Press
1963
7 Hawkes, H A . The Ecology of Sewage
Bacteria Beds (in Ecology and the
Industrial Society) John Wiley & Sons,
New York, pp 119-148 1965
Calaway, W.T. and Lackey, J.B
Waste Treatment Protozoa,
Flagellata University of Florida,
College of Engineering, Florida
Engineering Series No. 3, pp 1-140
1962
Calaway, W T The Metazoa of Waste
Treatment Processes-Rotifers.
Journal Water Poll Cont. Fed.
4(11) part 2 pp.412-422
Bick, Hartmut An Illustrated Guide to
Ciliated Protozoa used as Biological
Indicators in Freshwater Ecology
World Health Organization. Geneva
1969 (includes an illustrated key)
Curds, C. R. An Illustrated Key to the
Freshwater Ciliate Protozoa
commonly found in Activated Sludge.
Water Research Tech. Paper 12
Water Poll Res Lab. Stevenage
1969.
Curds, C. R and Cockburn, A.
Protozoa in Biological Sewage
Treatment Processes - I A
Survey of the Protozoan Fauna of
British Percolating Filters and
Activated Sludge Plants. II. Protozoa
as Indicators in the Activated Sludge
Process. Water Research
4-225-244 1970.
8 McKinney, Ross E and Gram, Andrew
Protozoa in Activated Sludge.
Sew Ind Wastes 28 1219-1231
1956 (reprinted in Biology of Water
Pollution by L. E Keup, W.M.
Ingram and Kenneth M Mackenthun
FWPCA Pub No CWA-3, pp. 252-
262 1967 )
9 Cooke, William Bridge, Trickling Filter
Ecology 40 273-291, pp. 269-287.
1959 (reprinted in Biology of Water
Pollution)
This outline was prepared by S. L. Chang,
M.D., Chief, Etiology, Division of Water'
Supply Programs Division, WPO, EPA.
Revised by R. M. Sinclair, Aquatic
Biologist, National Training Center, DTTB,
MDS, WPO, EPA, Cincinnati, OH 45268.
-------
Biota of Wastewater Treatment Plants
a. Peranema
trichoporum, 25u
d. N. gruberi, cyst
XT nuclei) 12u
b. Naegleria gruberi
amoeba stage, 18u
c. N. gruberi
flagellate stage
e. N. gruberi, cyst
(1 nucleus)
f. Endamoeba histolytic
cyst (4 nuclei) 16u
h. Actinosphae num sp
200-300u
l. Bodo caudatu
(with a navicula inside)
10-20u
g. Arcella vulgaris
40u
Fig. 1
S.L.Chang, 1963
-------
Biota of Wastewater Treatment Plants
a. Paramecium caudatum
200 - 260u
b. P. caudatum
cyst
c. Colpoda sp. 20-120u
Side view
Top
view
e. Euplotes carcinatus
70u
d. Colpoda cyst
Fig. 2
f. Vorticella 35-157u
S.L.Chang, 1963
-------
Biota of Wastewater Treatment Plants
a. Diaptomus sp. 2 mm.
(2 egg sacs
b. Cyclops sp. 2 mm.
Philodina sp. 45u
e. Diplogaster nudicapitatus
about 1 mm.
d. Anurea cochlearis 125u
Fig. 3
S.L.Chang, 1963
-------
Biota of Wastewater Treatment Plants
Hydrochna sp.
(50X)
(water mite)
Oorsal view
b. Folsomia fimetaria (50X)
c. F. fimetaria (side view)
d. Typical zoological organisms e. Typical zoological organisms
found in growth mass in found in floe masses in activated
Trickling filters (50X) sludge process (50X)
Fig. 4 S. L. Chang, 1963
-------
CJ1
I
a)
RHABIDITIS (Molt)
PHARYNX Oil
RECTAL GLANO
(Aft* Chitwood)4
ihwOGASTER (Molt)
OESOPHAGEAL
BULB
IRNACULUM
ICULES
MONHYSTERA (F«mol«)
UTERUS
CAUOAL CAMAL
OVARY
RHA6I0ITIS (Ftmalt)
I
UTERUS
SLANO
OESOPHAGEAL
(Aft** Cfcttwood)
Free Living Nematodes
-------
BIOLOGY OF ZOOPLANKTON COMMUNITIES
I CLASSIFICATION
A The planktoruc community is composed of
organisms that are relatively independent
of the bottom to complete their life history.
They inhabit the open water of lakes
(pelagic zone). Some species have inactive
or resting stages that lie on the bottom
and carry the species through periods of
stress; e. g., winter. A few burrow in
the mud and enter the pelagic zone at night,
but most live in the open water all the
time that the species is present in an active
form.
B Compared to the bottom fauna and flora,
the plankton consists of relatively few
kinds of organisms that are consistently
and abundantly present. Two major cat-
egories are often called phytoplankton
(plants) and zooplankton (animals), but
this is based on an outmoded classification
of living things. The modern tendency is
to identify groupings according to their
function in the ecosystem: Primary pro-
ducers (photosynthetic organisms), consumers
(zooplankton), and decomposers (hetero-
trophic bacteria and fungi).
C The primary difference then is nutritional,
phytoplankton use inorganic nutrient
elements and solar radiation. Zooplankton
feed on particles, much of which can be
phytoplankton cells, but can be bacteria or
particles of dead organisms (detritus)
originating in the plankton, the shore
region, or the land surrounding the lake.
D The swimming powers of planktonic
organisms is so limited that their hori-
zontal distribution is determined mostly
by movements of water. Some of the
animals are able to swim fast enough that
they can migrate vertically tens of meters
each day, but they are capable of little
horizontal navigation. At most, some
species of crustaceans show a general
avoidance of the shore areas during calm
weather when the water is moving more
slowly than the animals can swim. By
definition, animals that are able to control
their horizontal location are nekton, not
plankton.
E In this presentation, a minimum of clas-
sification and taxonomy is used, but it
should be realized that each group is
typified by adaptations of structure on
physiology that are related to the plank-
tonic mode of existence. These adapta-
tions are reflected in the classification.
II FRESHWATER ZOOPLANKTON
A The freshwater zooplankton is dominated
by representatives of three groups of
animals, two of them crustaceans*
Copepoda, Cladocera, Rotifer a. All have
feeding mechanisms that permit a high
degree of selectivity of food, and two can
produce resting eggs that can withstand
severe environmental conditions. In
general the food of usual zooplankton pop-
ulations ranges from bacteria and small
algae to small animals.
B The Copepoda reproduce by a normal
biparental process, and the females lay
fertilized eggs in groups which are carried
around in sacs until they hatch. The
immature animals go through an elaborate
development with many stages. The later
stages have mouthparts that permit them
to collect particles. In many cases, these
are in the form of combs which remove
small particles by a sort of filtration
process. In others, they are modified to
form grasping organs by which small
animals or large algae are captured
individually.
C The Cladocera (represented by Daphnia)
reproduce much of the time by partheno-
genesis, so that only females are present
Eggs are held by the mother in a brood
chamber until the young are developed far
enough to fend for themselves. The newborn
animals look like miniature adults, and do
not go through an elaborate series of
developmental stages in the water as do
the copepods. Daphnia has comb-formed
filtering structures on some of its legs
that act as filters.
BI.AQ. 29.5. 71
-------
Biology of Zooplankton Communities
D Under some environmental conditions the
development of eggs is affected and males
are produced. Fertilized eggs are produced
that can resist freezing and drying, and
these carry the population through
unsatisfactory conditions.
E The Rotifera are small animals with a
ciliated area on the head which creates
currents used both for locomotion and-for
bringing food particles to the mouth. They
too reproduce by parthenogenesis during
much of the year, but production of males
results in fertilized, resistant resting eggs.
Most rotifers lay eggs one at a time and
carry them until they hatch.
Ill ZOOPLANKTON POPULATION DYNAMICS
A In general, zooplankton populations are at
a minimum in the cold seasons, although
some species flourish in cold water. Species
with similar food requirements seem to
reproduce at different times of the year or
are segregated in different layers of lakes.
B There is no single, simple measurement
of activity for the zooplankton as a whole
that can be used as an index of production
as can the uptake of radioactive carbon for
the phytoplankton. However, it is possible
to find the rate of reproduction of the species
that carry their eggs. The basis of the
method is that the number of eggs in a
sample taken at a given time represents
the number of animals that will be added
to the population during an interval that
is equal to the length of time it takes the
eggs to develop. Thus the potential growth
rate of the populations can be determined.
The actual growth rate, determined by
successive samplings and counting, is less
than the potential, and the difference is a
measure of the death rate.
C Such measurements of birth and death rates
permits a more penetrating analysis to be
made of the causes of population change
than if data were available for population
size alone.
D Following is an indication of the major
environmental factors in the control of
zooplankton.
1 Temperature has an obvious effect in
its general control of rates. In addition,
the production and hatching of resting
eggs may be affected.
2 Inorganic materials
Freshwater lakes vary in the content
of dissolved solids according to the
geological situation. The total salinity
and proportion of different dissolved
materials in water can affect the pop-
ulation. Some species are limited to
soft water, others to saline waters, as
the brine shrimp. The maximum pop-
ulation size developed maybe related
to salinity, but this is probably an
indirect effect working through the
abundance of nutrients and production
of food.
3 Food supply
Very strong correlations have been
found between reproduction and food
supply as measured by abundance of
phytoplankton. The rate of food supply
can affect almost all aspects of pop-
ulation biology including rate of indi-
vidual growth, time of maturity, rate
of reproduction and length of life.
4 Apparently in freshwater, dissolved
organic materials are of little nutri-
tional significance, although some
species can be kept if the concentration
of dissolved material is high enough.
Some species require definite vitamins
in the food.
5 Effect of predation on populations
The kind, quantity and relative pro-
portions of species strongly affected
by grazing by vertebrate and inverte-
brate predators. The death rate of
Daphnia is correlated with the abun-
dance of a predator. Planktivorous
fish (alewives) selectively feed on
larger species, so a lake with alewives
is dominated by the smaller species of
crustaceans and large ones are scarce
or absent.
6 Other aspects of zooplankton
Many species migrate vertically con-
siderable distances each day. Typically,
migrating species spend the daylight
hours deep in the lake and rise toward
the surface in late afternoon and early
evening.
Some species go through a seasonal
change of form (cyclomorphosis) which
is not fully understood It may have an
effect in reducing predation.
-------
Biology of Zooplankton Communities
REFERENCES
1 Baker, A. L. An Inexpensive Micro-
sampler. Limnol. and Oceanogr.
15(5): 1,58-160. 1970.
2 Brooks, J. L. and Dodson, S. I.
Predation, Body, Size, and Com-
position of Plankton. Science 150:
28-35. 1965.
3 Dodson, Stanley I. Complementary
Feeding Niches Sustained by Size-
Selective Predation, Limnology
and Oceanography 15(1): 131-137.
4 Hutchinson, G. E. 1967. A Treatise
on Limnology. Vol. II. Introduction
to Lake Biology and the Limnoplankton.
xi + 1115. John Wiley & Sons, Inc. ,
New York.
5 Jossi, JackW. Annotated Bibliography
of Zooplankton Sampling Devices.
USFWS. Spec. Sci.: Rep.-Fisheries.
609. 90 pp. 1970.
7 Lund, J. W. G. 1965. The Ecology of
the Freshwater Plankton. Biological
Reviews, 40:231-293.
8 UNESCO. Zoolplankton Sampling.
UNESCO Monogr. Oceanogr. Methodol.
2. 174 pp. 1968. (UNESCO. Place
de Fortenoy, 75, Paris 7e France).
6 Likens, Gene E. and Gilbert, John J. This outline was prepared by W. T. Edmondson,
Notes on Quantitative Sampling of Professor of Zoology, University of
Natural Populations of Planktonic Washington, Seattle, Washington.
Rotifers. Limnol. and Oceanogr.
15(5): 816-820.
-------
Biology of Zooplankton Communities
FIGURE 1 SEASONAL CHANGES OF ZOOPLANKTON IN LAKE ERKEN, SWEDEN
Keratella cochleans
Keratella hicmalisi
Kellicottia longisplna
Polyarthra vulgaris
Diaptomus Rraciloides
Daphnia longispina
Cerlodaphnia quadrangula
Bosmina coregoni
N
D
J
A
S
0
M
A
M
J
J
P
1957
Each panel shows the abundance of a species of animal. Each
mark on the vertical axis represents 10 individuals/liter.
Nauwerck, A. 1963. Die Beziehungen zwischen Zooplankton und
Phytoplankton lm See Erken. Symbolae Botanicae Upsaliensis, 17:1-163.
-------
FIGURE 2 REPRODUCTIVE RATE OF ZOOPLANKTON AS A FUNCTION OF ABUNDANCE OF FOOD
0.20
0.15
0.10 -
0.05
Temperature
more than 10
/
Temperatu
-e
/ /
less than
10°
1
1
Young per
Brood —
20 -
10
0 200 400 600
Abundance of food organisms ygra/1, dry weight
Mean rate of laying eggs by the planktonic
rotifer Keratella cochlearis in natural
populations as a function of abundance of
food organisms and temperature. W. T.
Edmonson. 1965. Reproductive rate of
planktonic rotifers as related to food
and temperature in nature. Ecol. Mmogr.
35. 61-111.
Cells/ml
of food
100,000
Total young
Days
T
40
Number of young produced in each brood by Daphnia living in
four different concentrations of food organisms, renewed
daily. The total number produced during the life of a
mother is shown by the numbers at the right. The Daphnia
at the two lowest concentrations produced their first batch
of eggs on the same day as the others, but the eggs degen-
erated, and the first viable eggs were released two days
later. Richman, S. 1958. The transformation of energy by
Daphnia pulex. Ecol. Monogr. 28: 273-291.
c
3
(D
-------
Biology of Zooplankton Communities
PROTOZOA
Difflugia
Amoebae
Codonella
Stentor
Epistylis
Ciliates
-------
Biology of Zooplankton Communities
ROTIFERA
'///l /1
¥/.
Synchaeta
Polygarthra
Brachionus
Cladocera
ARTHROPODA
Crustacea
Copepoda
Nauplius larva of copepod
Insecta - Chaoborus
-------
Biology of Zooplankton Communities
PLANKTONIC BIVALVE LARVAE
86p.
377m.
xmple (gill attached)
(fin attached)
Glochidia (Unionidae) Fish Parasites
(1-3)
veliger
Veliger Larvae (Corbiculidae) Free Living Planktonic
(4-5)
Pediveliger attaches byssus lines)
Y
-------
MACRO INVERTEBRATES
I INTRODUCTION
Groups included arc in general those which
may be seen and recognized without the use
of a microscope For a more restricted
definition in reference to bottom sampling,
they are defined as those invertebrates re-
tained on a No. 30 sieve (approx. 0.5 mm
aperture).
II PHYLUM PORIFERA - Sponges
A Often encountered in the "pipe-moss"
complex. Being true animals, they will
grow in the dark and hence require only
a water possessing adequate food materials.
B Freshwater sponges usually appear as
brownish or greenish masses, (where
containing zoochlorellae), irregular in
shape, growing on twigs or solid surfaces.
Erect or branching shapes sometimes
found. Surface non-shiny, texture delicate.
May form overgrowths on irrigation
canal walls.
C Microscopic structure characterized by
silica "spicules" and reproductive
structures known as "gemmules"
1 Spicules are in general long slender
crystals In some, the ends are simple
or pointed, in others expanded into
various shapes
2 Spicules of various types are interwoven
like the twigs of a bird nest to form the
skeleton of the sponge The nature of
the soft living tissue cells indicate a
probable evolutionary origin of the
sponge from flagellated and amoeboid
protozoa
3 Gemmules are little bundles of tissue
cells, protected by spicules, which can
resist unfavorable conditions They are
often found scattered throughout the
mass of a sponge These are analagous
to turions in some aquatic plants and
statoblasts in bryozoans.
III PHYLUM COELENTERATA - The Jellyfishes,
Corals, and Hydras
A This group is of relatively little importance
in fresh water, although quite prominent
in the ocean.
B The freshwater hydra is a typical and
simple coelenterate Its structure is
essentially a long slender sac, two layers
thick, with tentacles extending out from
around the open end or mouth
1 Fully extended individuals may measure
a half inch or more
2 Generally an indicator of clean water.
Rarely a nuisance
C Craspedacusta, a freshwater jellyfish
occasionally appears in great numbers
in lakes and reservoirs in late summer.
No particular nuisance conditions are
known to result, although tastes and odors
might be expected if sufficient numbers
were taken into a water treatment plant
No reasons are known for their sporadic
appearances, no public health significance
is known, and no control measures can be
recommended.
D Cordylophora, a colonial hydroid is
a typical attached organism in large
rivers, lakes, and reservoirs.
IV PHYLUM PLATYHELMINTHES -
Tapeworms, Flukes, and Planarias
A Tapeworms and flukes are serious human
parasites in many parts of the world, but
generally under good control in North
America. Most of them have relatively
complicated life histories involving one
or more "intermediate" hosts and a "final"
host
BI.AQ 16d. 5. 71
-------
Macro Invertebrates
B The human tapeworm Diphyllobothrium
latum is a form of modest significance,
endemic to our northern states.
1 It is the largest of the human tapeworms
2 It is obtained by eating underdone fish
of the pike family
C Human fluke parasites have certain species
of snails as intermediate hosts Larval
forms, cercariae released from
snails penetrate human skin directly
while the person is wading or bathing in
infested waters.
1 Although one or two species of snail in
southern United States are thought to be
capable of transmitting the human blood
fluke, none are known to do so at the
present time.
2 Flukes also parasitize other animals
than man. Occasionally the cercaria
larvae of non-human parasites will be
attracted to humans bathing in infested
waters. They are able to enter the skin
but cannot complete penetration, and so
are trapped. The result is a rash, often
quite painful, known as "swimmer's
itch". This now occurs widely across
the northern states and in many coastal
waters. Control measures are directed
at the elimination of species of snails.
D The Planarians (Class Turbellaria) are
large enough to spot with the naked eye.
They are useful field indicators of pollution,
but shrivel up on preservation, and are
seldom recognized in the laboratory.
V PHYLUM BRYOZOA - Moss Animals
A These a^e small, colonial, sessile animals,
common in both marine and fresh waters.
Their main significance is as contributors
to the pipe-moss complex, and as indicators
of the degree of pollution.
B Freshwater forms are usually either
creeping brown moss-like forms that
grow over the undersides of rocks or
in pipes, or larger gelatinous masses
growing on sticks and rocks in lakes and
reservoirs.
1 The closely adhering threads on rocks
or pipes may range up to 1 /16th of an
inch in width. Microscopic examination
reveals numerous raised openings from
which when undisturbed, tiny fans of
tentacles (the lophophore) are extended.
Closely packed colonies may reach an
inch in depth, and if adjacent colonies
have come into contact, an indefinite
area may be covered.
2 Another type produces clear jelly-like
masses of transparent or faintly tinted
material, with minute, often colored,
individuals (zooids) scattered over the
surface. The animals are extremely
timid and only with great care and
patience can they be observed with the
tentacles expanded. Colonies of some
species are reported to approach six
feet in length, but smaller forms are
more common. Certain types of
colonies can slowly change their position.
3 Reproduction is by means of unique
structure known as a "statoblast".
This is a discoid or eleptical structure,
often with anchor-shaped hooks, which
can resist winter conditions and even
drying. Statoblasts usually germinate
in spring, colonies reach full develop-
ment by late summer.
VI PHYLUM MOLLUSCA - Snails, Clams,
also Oysters, Squids, Octopi
A Freshwater molluscs are in general
animals with a soft body, encased in a
calcareous shell which may be single
(snails) or double (clams and mussels).
Marine forms such as the squid, nautilus,
octopus, slugs, and others, would require
additional qualification.
B In the snail group (Class Gastropoda) the
shell may be coiled in various ways, or a
simple tent-shaped secretion on the
animal's back. These animals possess a
distinct head, with a pair of contractile
tentacles, at the base of which are placed
the eyes.
1 The mouth is provided with a unique
flexible rasp-like structure, the radula.
Chitinous jaws too are usually present.
-------
Macro Invertebrates
2 The two main groups in freshwater
are the air breathers (Order Pulmonata)
and the water breathers (Order
Streptoneura). Since all Streptoneura
have a peculiar chitinous or calcareous
"trap door" called an operculum (used
for closing the shell) they are also called
the "operculate" snails (vs the
"nonoperculate" Pulmonata).
3 Many snails are classed as "nuisance
organisms".
a Snails are quick to take advantage of
organic enrichment. As pollution
eliminates predators, pulmonate
snails (such as Physa, Lymnea) thrive.
Trickling filters, polluted streams,
and similar locations are often nearly
choked with these organisms.
b Certain snails are also the inter-
mediate host for certain fluke
parasites as mentioned elsewhere,
and hence may constitute an important
link in the control of these parasites.
C The Bivalved Molluscs (Class Bivalvia)
have the body protected by two symmetrical,
opposing valves or shells, which are united
above by a flexible elastic tissue called
the "ligament, " which is also secreted by
the mantle.
1 They have no head. The foot is an
axe-shaped mass of muscular tissue
which may be extended and used to
drag the animal ahead. The shell is
secreted by two sheets of tissue called
the mantle.
2 They feed by straining particles out of
the water by means of two sets of lace-
like gills (ctenidia). They are thus
animated filters and when present in
significant numbers may contribute to
the reduction of turbidity with resulting
solids accumulation.
3 Certain thick shelled forms such as the
Unionidae formerly commercially
harvested for use in making pearl
buttons, are exported to Japan for
production of nuclei for the cultured
pearl industry.
4 Certain small types (family Sphaerndae)
such as Sphaerium the fingernail clam,
have been shown to tolerate considerable
organic pollution.
5 The Asian Clam, Corbicula. An exotic,
intermediate msize between the above
two families, is a serious pipe clogging
organism. Unlike the endemic families
it has planktonic larvae, hence, it's
nuisance potential.
[I PHYLUM ANNELIDA - The segmented
Worm Earthworms, Sludgeworms, and
Leeches (Sometimes regarded as separate
phyla)
A Class Oligochaeta, the earthworm-
sludgeworm group. Body clearly divided
into segments. Bristle like "setae" or
hairs present on most segments, are used
in locomotion; in some species may be
withdrawn beneath body surface.
1 Accurate identification requires simple
clearing procedures with the specimen
Although these worms are herma-
phroditic (having both sexes in the same
individual), many of the smaller forms
commonly reproduce by a type of a
sexual budding which produces chains
of two or more individuals.
2 Aquatic earthworms and sludgeworms,
like their terrestrial counterparts, feed
on the soil or mud in which they live,
and contribute very significantly to its
stabilization. Having hemoglobin in
their blood, some of them can tolerate
very low oxygen tensions. Like the
snails mentioned above, they thus thrive
m polluted conditions in the absence of
predators (ex Tubifex, Limnodrilus. )
Smaller types (ex. Aelosoma,
Chaetogaster) may abound in activated
sludge.
B Class Hirudmea - leeches
1 These organisms are essentially
ectoparasites of vertebrates, though
they may also feed on smaller annelids,
snails, insect larvae, and organic ooze.
They are characterized by the possession
of an anterior and posterior sucker
-------
Macro Invertebrates
disc, and the absence of setae. Eyes
if present, are located on the (smaller)
anterior or oral sucker, although
sensory cells are widely scattered over
the general body surface.
They are not known to be the vectors of
any human disease, although when
present m numbers, their blood sucking
habits give them a considerable
nuisance value.
Their tolerance for sewage pollution is
considerable and they are hence often
present in great numbers in polluted
streams.
VIII PHYLUM ARTHROPODA - The Jointed
Animals
A Characterized in general by paired jointed
legs on a body nearly always segmented,
and a chitinous exoskeleton. Three of the
major groups have freshwater represent-
atives The Crustacea, Arachnida, and
Insecta. The Insecta will be treated in a
separate section.
B Class Crustacea
1 Characterized by two pair of antennae,
respiration by means of blood gills
(or general body surface). The vast
majority of crustacea are aquatic.
Crabs and lobsters are well known
marine examples, water fleas and
copepods well known freshwater
examples. No freshwater species
approach the giant marine species for
size where the king crab, for example,
may have a leg spread of several feet.
2 A few specialized terms used frequently
in connection with the Crustacea are
defined below.
Head:
The anterior part of the
body containing the mouth.
Usually consists of two
or more fused segments,
each represented by a
pair of specialized mouth-
parts.
Thorax The major section of the
body behind the head.
Contains most of the body
organs and usually the
walking (or swimming)
legs.
Abdomen- The most posterior section
of the body. Contains the
anus and often gills. Is
seldom involved in loco-
motion except in swimming
forms.
Caphalothorax The fused head and thorax.
Carapace.
A ntenna
Biramous.
A fold of the body wall
or shell which usually
extends down over each
side of the thorax. May
cover the whole side or
only the bases of the legs.
Sensory appendages or
"feelers". Typically two
in number in the
Crustacea.
Two branched.
3 Subclass Branchipoda - phyllopods
These organisms have many pairs of
flattened appendages serving for both
locomotion and respiration.
The first three orders as listed below
tend to inhabit temporary pools, and so
are often good indicators of such water.
Life histories may often be completed
in 2-3 weeks. Occurrence is quite
sporadic. Many of them are tolerant
of highly saline or alkaline waters.
The Cladocera, the last order, is more
of an inhabitant of permanent bodies of
water. All are in general, plankton and
detritus feeders, often tolerant of high
organic content as long as aerobic
conditions are maintained.
a Order Anostraca - fairy shrimps.
Eleven to 17 pairs of thoracic
appendages, elongate, cylindrical
body without a carapace, eyes stalked.
-------
Macro Invertebrates
Ex Artemia (the brine shrimp),
General size range 15-30 mm.
extreme 5-100 mm. Swim grace-
fully on their backs.
b Order Notostraca - tadpole shrimps.
Forty - 60 pairs of thoracic
appendages, body depressed and
partly covered by a dorsal shield
like carapace. Eyes sessile. Size
up to 100 mm.
c Order Conchostraca - clam shrimps.
Ten - 28 thoracic appendages. Body
laterally compressed and completely
enclosed in a bivalved carapace which
is often relatively thin and marked by
successive lines of growth. Generally
favored by warmer water. Size 4-
16 mm.
d Order Cladocera - water fleas. Four
6 pairs of thoracic appendages. Body
laterally compressed, all except the
the distinct head usually enclosed in
a bivalved carapace. Second antenna
is branched, and used for locomotion.
Single cpmpound eye. Size 0.2-3.0 mm
or more. Common genera include
Daphnia and Bosmina.
1) This is a large and widely dispersed
group, a common component of our
plankton in nearly all types of water.
2) Generally parthenogenetic, until
unfavorable conditions stimulate
the production of males. Sexual
eggs result which can withstand
freezing and drying.
3) Species in general are very widely
distributed.
4 Subclass Ostracoda - seed shrimps.
Two or three pairs of thoracic appendages.
Body laterally compressed, and entirely
enclosed in a bivalve carapace. Fresh-
water and marine. Size 0.35-21 mm.
No growth lines on valves (cf.
Conchostraca). Over 1700 species
known, about 1/3 freshwater.
"Microscopic clams with legs".
a Widely distributed, clean to polluted
water. Generally free living except
for a few rare commensals.
b Pollution significance is not known.
5 Subclass Copepoda - copepods.
Five - 6 pairs of thoracic appendages,
the 1st 4 biramous. Body cylindrical,
divided into two sections (Cephalothorax
or metasome, and abdomen or urosome).
Some parasitic forms are greatly
modified. Locomotion by means of
2nd antenna, which is unbranched
(cf. Cladocera). Many virtually
transparent. Up to 3 mm.
a Distribution world wide, freshwater
and marine. One of most abundant
of animal plankton.
b Development includes a complex
series of growth stages. Eggs
carried over from year to year
in mud. Resist drying and freezing.
6 Subclass Branchiura - fish lice.
With suction cups on head appendages,
body strongly depressed, ectoparasitic
on fish. Sometimes considered to be an
order of the copepoda. Of primary
importance as fish parasites.
7 Subclass Malacostraca - (no collective
common name). Body usually consisting
of 20 segments (approximately 5 in head,
8 in thorax, 7 in abdomen) and 19 pairs
of appendages exclusive of eyes.
Approximately 30, 000 species known
nearly 800 in N. America. Of the
12 orders recognized, only 4 have
freshwater representatives.
a Order Mysidacea - oppossum shrimps.
Essentially a marine group, but
three species inhabit our fresh
waters. Superficially resemble
marine shrimps of commerce.
Carapace thin and does not com-
pletely cover thorax. Stalked
compound eyes extremely large.
Nektonic in nature, with thoracic
appendages adapted for swimming.
-------
Macro Invertebrates
1) My sis relicta inhabits deep cold
oligotropic lakes in northern
states east of Great Plains Up
to 30 mm Circumboreal.
2) Acanthomysis awatchensis occurs
in lakes, rivers, and brackish
estuaries of Pacific N W
3) Taphromysis louisianae
Gulf coast region, also brackish
water Up to 8 mm
b Order Isopoda - aquatic sow bugs or
pill bugs Some fifty freshwater
species represent approximately 5%
of all known species, many of which
are terrestrial as well as marine
Size 5-20 mm
1) Ovoid, flattened dorsoventrally
Most of the thoracic and abdominal
segments are unfused, giving the
animals a many-jointed appearance
Lateral extensions of each segment
and the absence of any large pro-
truding structures combine to give
an overall impression of an army
tank in life
2) Generally inhabit springs, brooks
and subterranean waters In the
north central states they are often
abundant in small polluted streams
that go dry in summer, of which
they are frequently almost the only
inhabitants.
c Order Amphipoda - scuds or
sideswimmers Chiefly a marine
group with about 50 American fresh-
water species Size 5-20 mm.
1) Body is laterally compressed, few
fused segments as in isopods
Eyes generally well developed
except in subterranean species.
2) Occur in a wide variety of
relatively unpolluted waters where
ample oxygen is present Generally
nocturnal Soft waters generally
favored, but Gammarus limnaeus
is common m hard waters and
Hyalella azteca is sometimes
found in alkaline and brackish
waters
3) Subterranean species are common
in cavernous areas, and hence
frequently appear in well waters.
4) Scuds serve as intermediate hosts
for a variety of parasites of
waterfowl, amphibians, and fishes,
but not so far as known for man
d Order Decapoda - freshwater
shrimps, crayfish; also marine
lobsters, and crabs
Only about 160 species of this huge,
essentially marine group, are found
in the fresh waters of N. America,
of which about 130 are crayfishes.
True freshwater crabs occur in
Mexico and in the West Indies and
one species has been reported in
Florida. Other marine crabs
occasionally invade fresh waters
for extended periods and some have
become essentially terrestrial.
Decapods are in general from the
Rocky Mountain region.
1) Decapod shrimps (prawns) can be
distinguished by the laterally
compressed rostrum. Commercial
freshwater prawn culture techniques
have been developed. Size 3-23 cm.
2) The crayfishes (crawdads, crabs)
are the predominant group of
freshwater decapods. Their
shell is heavier and the pincers
usually strongly developed.
There are many burrowing forms
and many species have rather
specialized habitat preferences.
Unfortunately, however, their
pollution significance has not
been worked out. In general it
can be said that they can tolerate
a considerable amount of pollution.
Size, (exclusive of antennae)-
15-130 mm.
-------
Macro Invertebrates
C Class Arachnida - spiders, scorpions,
ticks, and mites. The water mites
(Parasitegona) have become extensively
adapted to fresh waters, and these are
almost exclusively restricted to fresh-
water, there being very few marine and
no terrestrial forms. They are readily
recognized by their bright colors,
globular to ovoid shape, and clambering
and swimming habits. Other types of
Acan or mites found in marine and fresh-
water are in the Halacaridae and Oribatei.
These crawling types are sometimes
found in large numbers in activated sludge.
Size 0.4 - 3.0 mm.
1 Superficially resemble minute spiders,
but have no division into cephalothorax
and abdomen. All evident segmentation
has been lost. Four pairs of legs are
present in the adult stage.
2 Mites are carnivorous or parasitic,
feeding on aquatic invertebrates. Some
are commensal on mussels, and host
specific.
REFERENCES
1 Eddy, S. and Hodson, A. C. Taxonomic
Keys to the Common Animals of the
North Central States. Burgess
Publishing Company, Minneapolis.
162 pp. 3rd Edition. 1961.
2 Palmer, E. Lawrence. Fieldbook of
Natural History. Whittlesey House.
McGraw-Hill Book Company, Inc.
New York. 1949.
3 Pennak, R.W. Freshwater Invertebrates
of the United States. The Ronald Press
Company. New York. 1953.
4 Pratt, H.W. A Manual of the Common
Invertebrate Animals Exclusive of
Insects. The Blakiston Company.
Philadelphia. 1951.
5 Pimentel, Richard A. Invertebrate
Identification Manual. Reinhold.
151 pp. 1967.
6 Stewart, R. Keith, Ingram, W. M. and
Mackenthun, K. M. Water Pollution
Control. Waste Treatment and Water
Treatment Selected Biological
References on Fresh and Marine Waters.
FWPCA, WP-23. pp. 126. 1966,
7 Ward, H.B. and Whipple, G.C.
W. T. Edmondson, ed. Freshwater
Biology. John Wiley & Sons. New York.
1959.
This outline was prepared by H.W. Jackson,
Chief Biologist, National Training Center,
WPQ EPA, Cincinnati, OH 45268 and
revised by R.M. Sinclair, Aquatic Biologist,
National Training Center
-------
Macro Invertebrates
3/k
Flatworms
Phylum PLaTYHELMINTHES
Planarla. a free living
flatworm, olass
Turbollarla
Van eats under-
cooked fish
Adult in
human liver
Plooo of fish
sporoovst
miraoidiura / \ la J
/ \Wi
/.
enoysted
oeroarla
3NAIL
— egg oontain
lag miraoldium
young redia
in sporooyst
/
oeroarla
Life history of human liver fluke,
Clonorohis sinensis. Class Tremefcoda
Nredia
young oeroarlae in redia
Aspeots in the life oyole of the human tapeworm
Dlphvllobothrlum latum, olass Cestoda. A, adult as in human intes-
tine; B,prooeroold larva in oopepod; C, plorooerooid larva in
flesh of piokerel(x-ray view).
H ,VT. Jaokson
-------
Macro Invertebrates
MINOR PHYLA
Phylum Coelenterata
Cordylophora caspia colony
Hydra with bud, \
extended, and contracted
Medusa of
Craspedacusta
Phylum Bryozoa
Massive colony on
stick
Statoblast
Creeping colony
on rock
Single zooid, young statoblasts in tube
-------
Macro Invertebrates
FRESH WATER ANNELID WORMS
Phylum Annelida mouth
anus
tubee
¦ mouth
Class Oligochaeta. earthworns
Ex: TubIfex the aludgeworm
(After Liebman)
u
m.
posterior sucker disc
Class Hirudinea, leeches
(After Hegner)
anterior end
Class Polychaeta . polychaet worms
Ex: Manayunkia, a minute, rare, tube
H V/. Jack son
PLATE XII c
building uorm.
(After Leidy)
-------
Macro Invertebrates
SOME MOLLUSC AN TYPES
Class: Cephalopoda*.
Squids, octopus,
cuttlefish.
Exclusively marine.
The giant squid shown
was captured In the
Atlantic In the early
nlnteenth century.
(After Hegner)
Llmax,
Lymnaea
Canpeloma
a slug an air breathing snail a water breathing
snail
Class: Gastropoda; snails and slugs. (After Buchebanm)
o '
-
\
© ^ "
' C
Class: Pelecypodaj clams, mussels, oysters.
Locomotion of a freshwater clam, showing how foot la extended, the tip
expanded, and the animal pulled along to Its own anchor. (After Buche-
baum) PLATE XII d H.W.Jackson
-------
Macro Invertebrates
3/4
Class CRUSTACEA
Crayfish, or orurdad;
Cambarua. Order Deoapoda
10—20 cm
Fairy Shrimp;
Eubranohlpus. Order
Phyllopoda
20-25 nun
Sow Bug; Acellua.
Order Xsopoda
10-20 mm
Tatar Flea;
Daphnla
Order
Cladooera
2-3 mm
8oud; Hvalella
Order Jkophlpoda
10-15 ™
Fish Louse, Argulus:
a parasltio Copepod
5-6 ma
H.W. Jaolc 6on
Copepod; Cvolops. irder Copepoda
2-5 ¦¦
PLATE X
-------
AQUATIC INSECTS
INTRODUCTION
The class Insecta of the phylum Arthropoda
includes approximately one million species,
more than all the rest of the animal kingdom
combined. Nine orders containing some
5000 species inhabit our North American
freshwaters. Representatives of these nine
orders will be discussed.
Adult insects possess a distinct head, a thorax
which bears three pairs of legs and one or two
pairs of wings, and an abdomen. The aquatic
dwelling immature stages are known as
larvae and nymphs. In some orders, the
immature stages closely resemble the adults,
while in others there is little or no resem-
blance. Larval insects that resemble the
adults mature through a series of "instars"
or "molts" called gradual metamorphosis.
Those that require changes from egg, larva,
pupa to adult undergo complete metamorphosis.
Some insects are able to live in an aquatic
environment because their respiratory
mechanism is adapted for life in water.
Oxygen is distributed to the tissues and CO^
removed by a system of air-filled tubes
called tracheae. Species that have access to
the atmosphere through the surface of the
water, or which can carry a bubble of air
below the surface can "breathe" air directly
and are independent of the dissolved oxygen
in the water.
Those larvae, pupae, and nymphs that cannot
"breathe" free air are sensitive to DO changes.
They must purify the tracheal air by exchange
with dissolved gases in the water. This is
done by means of specialized structures
known as tracheal gills and blood-gills.
Tracheal gills differ in their shape and posi-
tion on the animal. Sometimes they are
cylindrical filaments on the body, or flat
plates on the abdomen.
Also, considerable gaseous exchange is
thought to occur through the body's surface.
The responses of insects to organic pollution
is related to their respiratory needs and
capacities.
I ORDER PLECOPTERA (Stoneflies)
Figures 1-2
A The adults are relatively plain, colorless
insects of worldwide distribution. They
are generally poor fliers Each species
is rather definite in the time of its
appearance, and many species are most
abundant during the late fall or winter.
B The nymphs are all strictly aquatic and
relatively similar in structure. They
resemble the adults but have "wing pads"
instead of wings and two claws on each
tarsus. All nymphs have two tail filaments.
Filamentous tracheal gills are present in
some species, and as a group cannot
tolerate low DO's for extended periods.
II ORDER EPHEMEROPTERA (Mayflies)
Figures 3-8
A The adults possess delicate, veined,
transparent wings and two long tail
filaments. The mouthparts are undeveloped
and no food is taken during adult life.
Adults sometimes emerge in such numbers
near lakes and streams that they constitute
a distinct nuisance to man. Emergence
usually lasts only one or two days
B The nymphs live in a variety of habitats
in streams, ponds and lakes. They have
leaf-like tracheal gills attached to the
dorsal margins of their abdomens and
generally three slender tail filaments
posteriorly.
Ill ORDER TRICHOPTERA (Caddisflies)
Figures 9-15
A The caddisflies are all aquatic. The brown
colored adults distinctively fold their
wings so as to appear triangular in out-
line when resting. Generally nocturnal,
adult flights may create nuisance conditions.
They seldom live over 30 days.
BI. AQ. 17d. 5. 71
-------
Aquatic Insects
B Larva caddisflies live in protective cases
made from debris, sand, or pebbles.
Some are portable and others are fixed to
stationary structures. Their cases may
cover submerged rocks by the thousands.
C Although abdominal filamentous gills are
present, their DO requirements cover a
wide range and may be quite abundant m
organically enriched water,
IV ORDER ODONATA (Dragonflies and
Damselflies) Figures 17-21
A Adult dragonflies and damselflies are
large conspicuous foragers of almost any
body of water. They can be seen grace-
fully, flying just above the water's surface
in search of other insects. They hold
their wings horizontal even when resting.
B Odonate nymphs have a distinctive food-
grabbing device located under their mouth
used to capture passing prey. Some lie
concealed, others crawl about.
1 Dragonfly nymphs have unique enlarge-
ment of hind intestine, lined with rows
of tracheal gills that are not visible
externally. Water is kept moving by
"breathing" pulsations of abdomen and
the expelled water can aid in propulsion.
C Damselfly larvae on the other hand have
three leaf-like external tracheal gills
located on the rear of the abdomen. These
are vertical to the body and used to propel
the nymph.
D The ability of these nymphs to migrate
rapidly from one location to another makes
their value as pollution indicators doubtful.
V ORDER HEMIPTERA (True Bugs)
Figures 22-26
A Hemiptera are readily distinguished by the
combination of piercing and sucking
mouthparts, the anterior wings are
leathery at the base and membranous
apically while the hind wings are entirely
membranous. Most hemipterans are
terrestrial. Of the aquatic members,
many live on or near the water's surface.
The change-over from larva to adult is
gradual.
B No tracheal gills are known in the order.
Air is carried beneath the surface under
the wings or m masses of fine hairs.
Some Hemiptera, such as water boatmen,
are commonly found in polluted water.
Surface dwellers, such as water stridere
are quickly immobilized by a film of oil.
VI ORDER MEGALOPTERA (Dobsonflies,
Alderflies, Hellgrammites) Figures 27-29
A The adults are dull colored, up to 70 mm
in length. Although widely distributed,
they are seldom found in large numbers
and little is known about theu: life history.
B The larvae are among the most striking
of aquatic insects. The body is stout,
elongated, and up to 90 mm in length. A
pair of lateral, cylindrical, fleshy gill
filaments are located on each abdominal
segment. The legs are well developed
and the jaws strong and conspicuous.
VII ORDER LEPIDOPTERA (Aquatic Moths)
A Only members of a single family (out of
150) are known to have aquatic larvae.
The adults are small dull colored moths.
B The larvae are widely distributed over
U.S. in ponds with heavy growth of water-
lilies and similar vegetation. Their bodies
are whitish except for the dark head
capsules which bear strong jaws. Branching
filamentous gills are present on the
abdomen. The legs are reduced to fleshy
protuberances (prolegs).
VIII ORDER COLEOPTERA (Beetles)
Figures 30-34
A The adult beetle form is well known.
Horny or leathery covers represent the
-------
Aquatic Insects
front of two pairs of wings. Hind wings
are membranous and folded underneath.
Mouthparts are for biting. Some adults
are not structurally modified for aquatic
life (riffle beetles) and crawl about on the
stream bottom. In others the hind legs
may be greatly modified for swimming.
Most are dependent on atmospheric
oxygen that must be periodically replen-
ished at the surface.
B Some larvae are predacious and others
herbivorous. The head, thorax, and
abdomen are usually distinct and chitinized.
They respire by means of tracheae, lateral
gills, or anal gill tufts. The group, as a
whole, has a wide range to pollution.
IX ORDER DIPTERA (True Flies, Mosquitoes,
and Midges) Figures 35-39
A The adults are small delicate insects and
may be observed in swarms near a body
of water. They have one pair of functional
wings and a vestigial pair (reduced in size).
B The larvae are found in a variety of aquatic
habitats, and may attain enormous numbers
if tolerant to organic pollution. Although
the larvae are typically cylindrical and
wormlike, there are many morphological
adaptations for different modes of life.
The head may or may not be well developed.
Some members of the Chironomidae
(Midges) have an accessory respiratory
mechanism erythrocruorin (haemoglobin)
in the blood to permit life in low DO's.
These "bloodworms" also may possess
cylindrical blood-gills on the eleventh
body segment.
REFERENCES
1 Hynes, H.B.N. The Ecology of Stream
Insects. Annual Review of Entomology.
15:25-42. 1970.
2 Hynes, H.B.N. The Ecology of Running
Waters. Univ. of Toronto Press.
555 pp. 1970.
3 Fremling, Calvin. Mayfly Distribution
as a Water Quality Index. Water
Poll. Cont. Res. Series. 16030 DQH
11/70, EPA. Office of Res. and Dev.
Washington, D. C. 39 pp. 1970.
4 Needham, James G. and Lloyd, J. T.
The Life of Inland Waters. Comstock
Publ. Col,,Ithaca, N.Y. 438 pp. 1937.
5 Needham, James G. and Needham, Paul R.
A Guide to the Study of Freshwater
Biology, 5th ed., rev. Holden-Day,
San Francisco, pp. 108. 1962.
6 Pennak, R.W. Freshwater Invertebrates
of the United States. Ronald Press,
New York. 1953.
7 Usinger, R. L. Aquatic Insects of
California. Univ. of Cal. Press,
Berkeley. 1956.
8 Ward, H. B. and Whipple, G. C.
(W. T. Edmondson, ed.) Freshwater
Biology. John Wiley & Sons, Inc.,
New York. 1959.
C Pollution tolerance in the Diptera has been
associated with certain species. The group
is being used increasingly by biologists to
evaluate the effects of pollution on aquatic
life.
This outline was prepared by H. W. Jackson,
Chief Biologist, National Training Center,
Water Programs Operations, EPA, and revised by
W. T. Mas on, Jr., Aquatic Biologist, Analytical
Quality Control Laboratory, WPO, EPA.
-------
Aquatic Insects
Stoneflies
Order PLECOPTERA
(Isoperla confusa)
.antenna
.maxillary palpue
.lab rum
w
ocellus
wing pad
cercua
Adult
Figure 1
HWJ
-------
Aquatic Insects
May Flies
Order EPHEMEROPTERA
HWJ
Adult mayflies
(Heptagenlld&e)
Figure 3
Bymph of
Baetig
Figure 6
lymph of
Stenonema
Figure 4
Young njnnph of
Stenonema
Figure 5
Isonychla njnnph;
JL,8ide view; B, dorsal
Tiev of tail.
Figure 7
_l£Ofl nymph NOTE- Only two tail
Figure 8 filaments
-------
Aquatic Insects
Caddis Flies
Order TRICHOPTERA
T
Trlaenodes
A larva and case, B,adult-
Figure 9
Helicopeyche
larva; B.caee.
Figure 14
Brachycentrous
Larva and .case.
Figure 10
Hydropsyche
A adult; B.lavra.
Figure 11
leptocella
Larva and caae.
Figure 12
Ochrotrichia
Larva and case.
Figure 13
h
H.W.J
Molanna larva in case
Figure 15
"Log cabin" type
case of Linnophilid
larva.
Figure 16
-------
Aquatic Insects
Dragon Flies and Damsel Flies
Order ODONATA
Lateral aspect of damsel fly nympfc
Argia showing caudal tracheal gill a
Note labium folded under head.
Figure 17
Typical ^ adult
daiiisel fly
H lgure 18
Dorsal aspect of
Enallagma nymph,
a damsel fly.
Figure 19
A Nymph of the dragon fly Macromia snowing labium
extended (left) and redacted.
Figure 20
B. A typical adult dragon fly,
Macroaia- (2-3 in.)
Figure 20
Dragon fly nymphs Hagenms
(left) and Nasiaeschna.
Figure 21
-------
Aquatic Insects
True Bugs, or Sucking Insects
Order HEMIPTERA
Vl
Sucking beak
of typical
Hemipteran
Figure 23
Water Scorpion
(Ranatra spp.) Crawls about
in shallow water and elevates
"snorkel" tail to surface to
breath air.
Figure 22
Electric-light bug
(Lethocereua ap)
Extremely predaciouB on
small aquatic organisms. An
active flier positively
phototropic.
Figure 24
Water Strider
(Qerrie epp.)
^oerris spy.;
Jumps and glides about on
the sufface film.
Figure 25
I
Water Boatmen
(Corixa sp.)
This individual is shown with the
abdomen just piercing the surface
film to renew his air supply.
In well aerated water they are
able to remain submerged almost
Indefinitely.
Figure 26
HWJ
-------
Aquatic Insects
Alderflies, Dohsonflies, and
Hellgrammites
Order MEGALOPTERA (Neuroptera)
Alderfly Sialis
Adult
Figure 27
Figure 29 Dobsonfly Corydalus cornutus
A. larva (hellgrammite), B, adult
-------
Beetles
Order COLEOPTERA
upper
head
lower eye
1 diving "beetle (Dytlscus)
taking air at the surface.
Figure 31
I
Whirligig beetle (Qyrlnua) A. Side view
of head of adult shoving divided eye;
B. Larva; C( Adult. Carnivorous.
Figure 30
The Waterpenny (Psephenus);
A. adult; B, dorsal Bide of larva;
C, ventral side of larva. Predominantly
herbivorous.
Figure 32
A diving beetle (Cybister). The div-
ing beetles include some of the largest
and most voraaious of aquatic insects.
A, larva; B,adult.
Figure 33
Crawling water beetle;
A;adult; B,larva. Predominantly
herbivorous.,
Figure 34
HWJ
-------
Aquatic Insects
True Flies
Order DIPTERA
'J
'
nil
4
Midge Adult
Figure 36
A. sewage fly larva
(Psvchoda)
Sewage fly pupa
(Psychoda)
Midge pupa
(Chironomus)
1- igurc 35
HWJ
ldge larva
(Chironomus)
Figure 37
c Adult sewage fly
(Paychoda)
Figuie <8
Rat-tailed maggot
(Iristalle)
A, adult; B.larva.
Figure 39
-------
KEY TO ORDERS OF AQUATIC INSECTS
1 Without jointed thoracic legs;
maggot-like... Larvae of
DIPTERA (flies) Figure 37
1' Jointed thoracic legs present
Figure 2
2 Long, segmented appendages
at posterior end Figure 2 and 4
2' Posterior appendages absent, or
if present, not long and segmented
Figure 28 and 33 A
3 Two posterior appendages, two
tarsal claws, usually with
finger-like tracheal gills on
ventral side of thorax. Nymphs
of PLECOPTBRA (stoneflies)
Figure 2
3r Usually three (sometimes two)
posterior appendages; one tarsal
claw, tracheal gills on lateral
margins of abdominal segments.
Nymphs of EPHEMEROPTERA
(mayflies) Figure 4
4 Wings or wing pads present,
nymphs and adults Figure 24
4' Without wing pads; larvae
Figure 28
5 Wings, hard or chitinous (elytra),
hind wing may be complete or
vestigial. COLEOPTERA (beetle
adults) Figure 33B
5' Wing pads present, nymphs
Figure 21 and 26
6 A long jointed beak present.
Nymphs and adults of
HEMIPTERA (bugs) Figure 23
6' Without a long jointed beak;
labium, when extended, long
and scooplike, and when folded,
serving as a mask covering the
other mouth parts. Nymphs of
ODONATA Figure 20
7 Platelike caudal gills present;
damselflies. Figure 17
7' No platelike caudal gills;
dragonflies. Figure 21
8 Five pairs of abdominal prolegs.
LEPIDOPTERA (aquatic
caterpillars)
81 Prolegs absent, or confined to
last abdominal segment
9 Each abdominal segment with
one pair of stout lateral processes
MEGALOPTERA (hellgrammites,
alderfly and fishfly larvae)
Figures 28 and 29A
9' No stout lateral abdominal
processes; sometimes with
long, thin lateral filamentous
processes (a few beetle larvae
have four stout hornlike pro-
cesses on each body segment)
Figure 30B
10 A pair of terminal abdominal
prolegs, usually in fixed or
portable cases. TRICHOPTERA
(caddis fly larvae) Figure 11B
10' Without terminal prolegs.
COLEOPTERA (beetle larvae)
Figure 30B
10
This outline was prepared by John E. Matthews,
Aquatic Biologist, Robert S Kerr Water
Research Center, Ada, Oklahoma 74820.
BI. AQ. 28. 8. 69
-------
FRESHWATER CRUSTACEA
Part 1. Crustacean Diversity
I The two major classes of arthropods in
freshwater are crustacea and insects. The
diagnosis of the class Crustacea is simply,
"arthropods with two pairs of antennae. "
A Crustacea share these features:
1 The great majority of the 30, 000 known
species of crustaceans are marine.
Only a very few are terrestrial.
2 Two pairs of antennae. The first pair
are antennules.
3 Three pairs of masticating and feeding
appendages.
4 The paired appendages are typically
biramous (Figure 1)
Figure 1. Abdominal Ring of a Decapod
(from Schmitt)
TABLE 1
CLASSIFICATION
CLASS CRUSTACEA
"Microcrustacea" - Entomostraca)
Subclass Branchiopoda
Subclass Malacostraca
Order Anostraca
Division Pericarida
Order Notostraca
Order Mysidacea
Order Conchostraca
Order Isopoda
Order Cladocera
Order Amphipoda
Subclass Ostracoda
Division Eucarida
Subclass Copepoda
Order Decapoda
Order Calanoida
Order Harpacticoida
Order Cyclopoida
BI. ART. 4a. 5. 71
-------
Freshwater Crustacea (Part 1)
B Classification into two groups is admittedly
artificial.
1 Entomostraca, an old classification, is
used now as a general descriptive term
for the generally smaller sized
crustacea in eight diverse orders; often
called microcrustacea.
2 Malacostraca is a natural group, all
members possessing nineteen segments
(plus one embryonicl(Figure 2).
II CRUSTACEA AND MAN
A Crustacea are important as first order
consumers. The Cladocerans and Mysids
are essential in the food chain, a link
between the algae and many fish species.
B Commercial fisheries exist for crayfish
in Louisiana.
C The Sins of Crustacea
1 One group of copepods is parasitic upon
fish.
2 Amphipods are destructive to fish nets
in some lakes.
3 Case making amphipods seriously
contribute to the irrigation canal wall
growths which reduce flow.
Some copepods are intermediate hosts for
vertebrate helminth parasites.
Microcrustacea have been the cause of
spotting of paper in paper mills. Oligotrophic
lakes serving as a water supply for the mill
contained sufficient numbers of copepods to
seriously spot paper rolls. The pigment,
astaxanthin," within the cells of the copepod
turns red, leaving discolored spots in the
finished paper.
Ill Crustacean life history involves distinct
stages from egg to adult.
A The first larval stage is the nauplius,
plural nauplii (Figure 3).
1 The malacostra brood the eggs, often in
marsupia, thus naupliar stages are
passed in the egg membrane.
2 The "entomostraca" generally have
free swimming nauplii.
a Cladocera, with the exception of
only a few genera, do not have a
free-swimming nauplius stage.
b The great majority of nauplii encountered
in freshwater zooplankton are copepods.
Figure 2. Malacostracan Organization
(after Russell-Hunter)
Figure 3. Nauplius Larva
:o: ijctc ccrapoce
sin 3! 3
roifrum
pcirv>d
eyes
^ fir;!
ci: J.-vr.a
/
1 .1 -- • — *
crlorr.o 1 | $ pCi;rn thcfatie 'irnbs-
\ n.nm'ible j
\ fiinvriv:;;;? \ +— - in Qatapods' -
r-y.ft nurx.lla ? p::'n r.riAi'l.pacls
1 choice.
. , ; •vuir.I '-f; legs
In! son
.re yods
6 p'liii jodon'Ind limbs
-------
FRESHWATER CRUSTACEA
Part 2. Entomostraca
I As implied,the Anostraca are without a
shell or carapace
A They are the least specialized of the
Branchiopods.
1 Trunk limbs are all similar and used
both in swimming and feeding
2 They are filter feeders.
3 Types include
a Artemia, the brine shrimp
b Branchipus, living in brackish water
c Chirocephalus and other genera,
known as fairy shrimp, living in
ponds which dry up in the summer.
4 Size is generally up to two cm, but some
reach as much as ten cm as adults
B Artemia, the brine shrimp, is commercially
valuable.
1 The adults are sold frozen in bulk.
2 The dried eggs which can be stored in
diapause and hatched as needed are
widely used in fish culture.
II Notostraca or tadpole shrimp have a
carapace forming a dorsal shield which
covers only about half of the body.
A Only two genera are known, Triops and
Lepidurus.
B Triops has been a serious pest of rice
fields as it stirs up the bottom sediments,
killing the rice seedlings.
III The Conchostraca possess a bivalve shell,
completely enclosing body and limbs, re-
sembling the shell of a small clam.
IV Cladocera have a similar bivalve shell or
carapace except the head is left free The
second antennae are the main means of a
jumpy movement, hence the name water fleas
V Ostracoda have a bivalve carapace and
resemble Conchostraca except for the
absence of growth lines on the valves.
A Average size is 1 to 3 mm.
B They are found in all types of fresh water
C Large populations may develop in algal
cultures and aquaria. This habit is
detrimental to culture of aquatic snails.
VI The Copepoda, like the Cladocera, are
almost universally distributed in the plankton,
benthic, and littoral regions of fresh water
A Copepods in the Order Harpacticoida are
substrate dwellers, the other two orders
Cyclopoida and Calanoida are mainly
swimmers (Figure 4).
B Copepod eggs hatch into a small larva,
the nauplius, which has three pairs of
stubby bifurcate appendages, representing
the first antennae, second antennae, and
mandibles (see Figure 3).
C As in the Cladocera some common species
of planktonic copepods exhibit diel
verticle migrations in lakes and ponds, with
a greater concentration of individuals in the
upper waters during the hours of darkness
Q
Figure 4 a. Calanoid
b Cyclopoid
c. TIarpacticoid types
arrow marks hinge line
-------
FRESHWATER CRUSTACEA
Part 3. Malacostraca
I The subclass Malacostraca includes four
orders of crustacea, and contains almost
three quarters of all the known species of
crustacea.
A Characteristics of this group distinguishing
them from the six orders commonly termed
"Entomostraca" include-
1 Tagma (an organized group of segments
or somites forming a distinct body
section) 5-8-6.
2 A gastric mill in the stomach.
3 The larva is usually passed
within the egg, and when present, it
does not feed.
B The most primitive of this group are
filter feeders.
H ORDER MYSIDACEAE
A The mysids or opossum shrimp occur in
the marine plankton and over littoral
sand-flats; the few forms in fresh waters
are relatively recent marine relict species
as shown by physiological investigations.
B The last pair of pleopods are enlarged,
containing statocysts, being used as
rudders and elevators for the swiftly
darting mysids.
C Three freshwater species occupy different
areas of North America.
1 Acanthomysis awatchensis is found m
streams on the Pacific Coast.
2 Mysis r ell eta is found in the Great
Lakes and northward.
3 Taphromysis louisiar.ae is found in
streams of the Gulf Coast (Figure 5).
Figure 5. Mysidacea. Lateral view of
Taphromysis louisianae (c, carapace; t,
free thoracic somites). Modified from
Banner.
Ill ORDER ISOPODA
A The most striking characteristic of the
isopods is the dorsoventrally flattened
body (Figure 6) which is in contrast to
the lateral compression of most
amphipods. Thus most isopods crawl
at least part of the time. Lygia, a marine
isopod, can move each leg sixteen steps
per second. Other isopods can run back-
ward as rapidly as forward.
Figure 6
-------
Freshwater Crustacea (Part 3)
B The most common freshwater isopods
(log lice, sow bugs, water slaters) in
North America belong to the genera
Lirceus and Asellus (Order Asellota,
Family Asellidae).
1 These isopods typically form the well
known third peak below organic dis-
charges, e.g., Tubificidae, Chirono-
midae, Asellidae.
2 Lirceus. The lateral margins of the
head are produced to overhang the base
of the mandibles, the anterior edge of
head is produced (Figure 7).
Figure 7
Lirceus
3 Asellus. The above characters
negative.
Figure 8
Asellus
IV ORDER AMPHIPODA (Figure 9)
A Another difference between isopods and
amphipods, also concise and consistent
is in the thoracic and abdominal
appendages, each being arranged m at
least two groups. In the isopods all
thoracic limbs and all abdominal limbs
are similar.
Figure 9 Amphipoda. Lateral view of Gammarus
a, first antenna, u, urosome
B Most freshwater amphipods are substrate
oriented and active in swimming.
1 Pontoporeia is both benthic and nektonic,
a glacial relict of both Eurasia and
North America
2 Corophium spinicorne, a tube building
amphipod, is found on the West Coast.
It is a major contributor to the com-
munity lining irrigation canal walls.
V ORDER DECAPODA
Includes freshwater shrimp, crayfish and
crabs.
A Decapods are distinguished from all other
Malacostracans in that the first three
pairs of thoracic appendages are modified
as maxillipeds. The remaining five pairs
of thoracic appendages are legs, hence,
the name decapoda. The legs are chelate,
the first pair which is often heavier is
called a cheliped
B Two suborders are recognized.
1 Natantia (natant - "swimming"), the
body generally adapted for swimming
and laterally compressed. The rostrum
is keeled(Figure 10). This suborder
includes the freshwater shrimp.
2 Reptantia (reptant - "crawling"), the
body generally adapted for crawling
and dorso-ventrally flattened to a degree.
The rostrum is flattened dorso-
ventrally (Figure 11). This suborder
includes crayfish and only a few fresh-
water crabs.
Figure 10
Figure 11
-------
Freshwater Crustacea (Part 3)
REFERENCES
1 Bousfield, E. L. Freshwater Amphipod
Crustaceans of Glaciated North America.
Canad. Field-Nat. 72(2):55-113. 1958.
2 Hobbs, Horton H., Jr. The Crayfishes of
Florida. Univ. of Fla. Biol. Sci.
Series 3(2):1-179. 1942.
3 Holsinger, John R. Systematics,
Speciation, and Distribution of the
Subterranean Amphipod Genus
Stygonectes (Gammaridae). Bulletin
259. U.S. Natural Mus., pp. 176.
1967.
4 Schmitt, Waldo L. Crustaceans. Univ.
Mich. Press, pp. 204. 1965.
/
5 Segerstrale, Sven G. The Immigration
and Prehistory of the Glacial Relicts
of Eurasia and North America. Int.
Revue ges. Hydrobiol. 47 (l):l-25.
1962.
6 Waterman, Talbot H. The Physiology of
Crustacea, Vol. I. Metabolism and
Growth. 670 pp. 1960. Vol. II.
Sense Organs, Integration and Behavior.
681 pp. 1961.
7 Williams, W.D. A Revision of North
American Epigean Species of Asellus
(crustacea. Isopoda) Smithsonian
Contrib. to Zool. 49-1-80. 1970.
This outline was prepared by R. M. Sinclair,
Aquatic Biologist, National Training Center,
WPQ EPA, Cincinnati, OH 45268.
-------
FRESHWATER MOLLUSCA
Part 1. General Concepts
I In terms of number of individuals, number
of species, and energy flow through the
group, molluscs are clearly a major phyla
ranking only below the Arthropoda in this
respect.
A The term mollusc means soft, referring
to a soft body within a hard calcareous
shell.
B Extensive use is made of cilia and mucous
mechanisms, in feeding, locomotion,
respiration, and reproduction.
C The hard parts of calcareous shell have
long been used in systematics in this
group.
1 The gastropods or univalves have a
single shell spirally coiled in one or
more planes, or have a low cone
shaped shell, as m the limpets. In
both these groups the body is spirally
coiled and asymetrical.
2 The bivalves have two calcareous
valves united by an elastic hinge
ligament. The body is bilaterally
symetrical.
II Structure and function are homologous in
both gastropods and bivalves with some
exceptions.
A The soft body is characteristic.
Prominent features are-
1 The foot is used for locomotion -
gliding m many gastropods and
burrowing m bivalves and some
gastropods. As in other soft parts,
the foot is a remarkable hydraulic
skeleton or haemoskeleton which
allows for startling changes in shape.
2 Gills or ctenidia. This feature is
characteristic of the phylum mollusca
only.
B The shell is composed primarily of
calcium carbonate and thus, reflects
water chemistry.
C Reproduction
Typical stages following the egg are the
trochophore, veliger, pediveliger, and
juvenile. This basic plan is followed m
both orders, whether oviparous,
larviparus, or viviparus.
D Feeding is typically filter or suspension
feeding and browsing.
1 Gastropods possess a radula. In the
molluscs this is lacking only in the
bivalves. The radula is used for
rasping food from the substrate or
detritus. Some gastropods may also
suspension feed.
2 Bivalves are suspension or filter
feeders. They possess a crystaline
style (some gastropods also have this)
which revolves in a style sac taking
the food chain (a rope of mucous with
food particles) sorted by the palps, in
a windlass type operation down the food
canal. This is the only revolving organ
in nature. As it rotates against a
gastric shield, enzymes are liberated
for food digestion.
Synoptic Classification - Phylum Mollusca-
Class Gastropoda
1 Order Mesogastropoda
2 Order Basommatophora
Class Bivalvia
3 Order Schizodonta
4 Order Heterodonta
BI. MO. 4a 5. 71
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FRESHWATER MOLLUSCA
Part 2. Gastropoda {Snails and Limpets)
I The Class Gastropoda is represented in
virtually every aquatic habitat from cave
streams to swamps. All are substrate
oriented, and are found in lakes at depths
as great as 250 meters to shallow pools.
Each body of water has its characteristic
snail fauna. Although some groups, for
example, are referred to commonly as River
Snails (family Pleuroceridae) and Pond Snails
(Lymnaeidae), they are not so restricted.
II GENERAL CHARACTERS
The majority have a spiral (cone or discoidal)
The limpets have a shell shaped like a coolie
hat.
1 Spiral 2 discoidal 3 limpet
A The shell is composed of calcium
carbonate and covered by an organic layer,
the periostracum.
If the shell aperture is on the observers
right (coiled counter clockwise), it is
dextral and if to the left (coiled clock-
wise), it is sinistral.
4 DEXTRAL
5 SINISTRAL
2 The shell may show considerable
variation even within a single species
at a given location or in a sequence in
a river basin (Plate 1),
3 The major shell features are illustrated
in Figure 9.
spire
— columella
_ umbilicus
- aperture
— canal
L. £. geniculata
L. g, fuliginosa
Figure 9
L. jr. puiguis
river knobbed form 8 smooth headwater form
down stream form
Lithasia geniculata
Unfortunately, these many varieties were once described as separate species,
so many species today have a formidable synonomy. It is no less unfortunate
that the majority of today's species have been "lumped, " also intuitively.
PLATE 1
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Freshwater Mollusca
B The gilled snails (Streptoneura) have an
additional structure, the operculum (10-12),
a chitinous plate, which closes the aperture
when the body is withdrawn in the shell.
I/'
(' •
V
v \
- \
\ V
i '
' i
/
10 multispiral 11 concentric 12 paucispiral
C The body (often called "soft parts" in
contrast to the shell) is attached and
withdrawn into the shell by the columellar
muscle.
c Pseudobranch
This respiratory (modified gill)
structure is an external extension
from the foot and is found in the
families Ancylidae and Planorbidae
(in the Basommatophora).
3 Remaining parts of the body conform to
the internal cavity of the shell and con-
tain portions of the gut and also
reproductive systems.
4 Radula
A conspicuous part of the buccal mass
(anterior part of head) is the radula,
which is one of the most characteristic
features of the gastropoda. This toothed
structure is used for rasping food from
the substrate.
1 Foot
The muscular part of the anatomy
which protrudes from the shell is called
the foot. Typical structures of the foot
including the head are shown below
tentacle
¦ \V'WPr°boscis
operculum
Figure 13
2 Mantle
The thin sheet of specialized tissue
which lines the shell and also secretes
the shell material during growth is
called the mantle. Specialized mantle
areas include:
a An external gill or ctenidium com-
posed of a series of plates and
highly vascular. Only the order
Streptoneura have ctemdia.
b An internal lung cavity with an
external opening, the pneumostome
which enables them to breathe air
at the surface or circulate water and
extract the DO. All nonoperculate
freshwater snails (of the order
Basommatophora-Pulmonata) have
this structure.
Figure 14
The order Streptoneura have seven teeth
per row as shown.
The order Basommatophora, have more
than seven (19 to 175).
HI ECOLOGY
Distribution of snails is closely related
to water chemistry, physical characters of
the habitat, geography, and food habits.
A Chemistry
Distribution is related in large measure
to dissolved salts, especially calcium
carbonate, the essential material for shell
construction. Other factors include pH
which is associated with and partly
determined by the CO^ content.
B Zoogeography
The gastropod fauna in the United States
is very diverse and largely endemic. Such
genera as Lymnaea, Helisoma, Gyraulus,
and Ferrisia are widely distributee^ The
Pilidae, for example, are confined to
Florida and southern Georgia. Some
rivers and river systems have unique faunas.
-------
Freshwater Mollusca
1 An ecological principle is, "the more
extensive a species ecological range,
the wider its distribution will be 1
Examples are these introduced species
which in their home ranges have both
an extensive range and show continued
expansion in their original range.
a Melanoides tuberculata
Afroasian, now found in Texas and
Florida (intermediate host for the
liver fluke producing Clonorchiasis)
b Thiara granifera
Asian, now also found in Texas and
Florida (intermediate host for the
lung fluke producing Paragonimiasis.)
c Viviparus iapomca
Asian, now widely distributed in
United States, especially numerous
in Lake Erie,
d Bythinia tentaculata
European, now numerous in northern
states and the Great Lakes.
e The first three, Asian species, also
exhibit great divergence in shell
characters and as a result, these
varieties have been described as
species for each island and land mass.
D Food
Since most snails are browsers, they are
dependent upon periphyton and berthic
algae to a large degree
IV COLLECTION
The exploitation of many habitats, secretive
habits, and migrations make this group the
most difficult benthic invertebrate fauna to
collect, next to the bivalves. To put it another
way, one must know what species to expect,
the particular habitat to search, and the
appropriate collecting gear.
V PREPARATION
Although snails usually may be identified
without the body and/or the operculum, these
are sometimes essential. Therefore, these
animals should be relaxed, killed, fixed, and
preserved in toto. The first step is the most
difficult. A variety of relaxants including
Nembutol, menthol, and organic pesticides
have all been successfully used Unbuffered
formalin will eventually destroy all the
shell except the periostracum, thus shelf life
of preserved bottom samples is seriously
shortened. The remaining periostracum in
carelessly preserved collections will be a
crumpled mass and unidentifiable.
Exotics lacking parasitic controls
and competition often reach large
densities. For example, the first
two mentioned (a and b) have reached
populations in the San Antonio area
of 33 per square inch, and this density
was maintained over a period of
three years observation. (Murray
and Wopschall).
2 Due to changes in water quality, includ-
ing impoundment, a number of endemic
genera and species are either extinct
or are endangered because of their
limited ecological tolerances
C Water quality requirements of even our
common gastropods are still not com-
pletely understood. In general, however,
gilled snails are seldom found in areas of
gross organic pollution, while air breath-
ing snails may be found almost anywhere
(unless excluded by other factors). There
are so many exceptions to pollution and
gastropod relations that generalizations
are not meaningful.
20-4
VI Identification requires precise locality data for
the specimen at hand, well preserved material,
appropriate keys, and descriptive material for
confirming identification
A The gastropods are a relatively simple
group to key and have been carefully
studied, the great majority being known
for years.
1 A real problem here is the confusion
resulting from investigators unable to
resolve problems in accepted names.
a Pleurocera zonalis Raf. equals
Lithasia obovata Say
b Pleurocera canaliculata Say equals
Oxytrema canaliculata Say
2 In the Lymnaeidae the problem involves
generic and/or subgeneric classification
as well as what constitutes a species
3 For these reasons specific identification
should involve consultation with a
specialist in the particular family in-
volved. Then one should remember that
taxonomy is far from settled in these
-------
FRESHWATER MOLLUSCA
Part 3. Bivalves
I The great majority of living bivalves are
lamellibranchs, that is, they have enormously
enlarged gills. These ctenidia, like those in
a few genera of mesogastropods such as
Pleurocera, are enlarged by elongation of
the filaments so that, together, adjacent
filaments form a lamella.
A These lamellae are more than sufficient
for respiration. They are in fact the
major organ of food collection in these
filter-feeders.
B A water current through the mantle cavity
is created by the lateral cilia. This flow
passes between the filaments of the
ctenidium from the inhalant siphons to the
exhalant siphon. The particulate matter
remaining on the inhalant face of the gill,
the frontal cilia and mucus are used to
make chains or boluses of this material
that then pass on to the mouth.
C The amount of water passed through this
system is relatively large, for example, a
small 20 gm clam can pump nearly four
liters per day. Thus, clams concentrate
heavy metals, radionuclides, pesticides,
bacteria, and viruses, etc.
D Wastes from this system include feces
and pseudofeces.
1 Fecal deposits are discharged from the
anus into the exhalant siphon. These
tightly bound and mucous connected
strings may smother older generations
m a deposit. They also may contain
significant amounts of nutrients and/or
pesticides.
2 Pseudofeces are mucous connected
sheets which are rejected solid
materials collected on the gills in the
feeding process. By vigorous mantle
contractions of the shell these are dis-
charged through a mantle opening.
Pseudofeces may form significant benthic
deposits (phosphorus in biogeochemical
cycles) which alter irrigation canal
hydraulics. Pediveligers may also be
trapped in this mass.
II The structure of all five families, all
belonging in the same subclass of freshwater
bivalves is basically the same.
A Size and reproductive habits differ in
each family.
Table 1
PHYLUM MOLLUSCA - CLASS BIVALVIA
SUBCLASS EULAMELLIBRANCHIA
ORDER SCHIZODONTA
SUPER FAMILY UNIONACEA
1 FAMILY MARGARITIFERIDAE
2 FAMILY UNIONIDAE
ORDER HETERODONTA
SUPERFAMILY SPHARIACEA
3 FAMILY SPHAERIIDAE
4 FAMILY CORBICULIDAE
5 FAMILY MACTRIDAE
1 The mussels (Margantiferidae and
Unionidae) are classed in the super-
family Unionaceae.
a They exhibit schizodont tooth structure,
b They reach a size of up to 9 inches.
c Longevity is relatively long, up to
> 90 years.
d Many species are of commercial
value for shell export to Japan. The
shells being processed into nuclei
for production of cultured pearls in
the Japanese pearl oyster.
e Commercially valuable species have
accepted and widely used vernacular
names, such as pigtoe, maple-leaf,
monkey-face, pocketbook, etc.
f Unionids are obligate fish parasites.
The glochidia (homologous with
veliger larvae) are discharged from
the gills into the water, clamp onto
the gills or fins of the host fish.
After a short parasitic development
period, they drop off to begin benthic hfe.
-------
Freshwater Mollusca
Corbicula is intermediate in size and
longevity.
7
pseudo-
cardinal
teeth
umbo (beak)
ligament
lateral teeth
post add scar
pallial line
15 heterodont
16 schizodont
17 Interior and Top View ofMussel Shell
Fingernail clams (Sphaeriidae) and an
introduced species Corbicula
(Corbiculidae) have heterodont teeth
(cardinals and laterals) and belong to
the same superfamily, Sphaeriacea.
a Up to 65 mm in shell length
b Longevity is approximately five to
ten years.
c They are canal clogging, pipe clogging,
etc., organisms of serious magnitude.
d The veliger larvae are discharged to
the water in large numbers.
Pelagic larvae are then freely
carried into raw water systems. A
byssus and highly specialized foot
allows them to establish a sessile
habit in flowing conduits or form
loosely aggregated masses in the
substrate.
The remaining family, Sphaeriidae, are
the smaller and shortest lived group.
a Size is from two to twenty mm, but
usually not more than ten mm.
b Longevity is up to two years.
c The genera Sphaerium, Musculium,
and Eupera are nearly equipartite,
and thus are called "fingernail clams'.'
The genus Pisidium is inequipartite
and these are called "pea shells. "
d One to twenty individuals develop
within the marsupial gills. These
are released as juveniles.
Ill Shell and body are structurally and develop-
mentally a single entity although they are often
described as though the two valves and the
ligament are of different origin.
A Basic shell structures are as shown.
1 Attachment of muscles and other soft
parts are clearly marked on the shell
as impressions. The largest bivalve
muscles are the adductors which pass
through the mantle tissue and are
attached to each valve. The elasticity
of the ligament and several kinds of
hydraulic systems serve as antagonists
for the adductors.
2 Shell teeth are simply shell projections
which serve as a fulcrum for anticulation
of the shell valves.
3 The shell is composed of three layers
a The outer or periostracum is mainly
organic.
b The mid or prismatic layer is com-
posed of minute prism-like blocks of
calcium carbonate.
c The inner or nacre (mother of pearl)
is composed of alternating layers of
calcium carbonate and an organic
substance. The species with the
thickest nacre and silver-white sheen
were originally the most valuable for
the pearl button industry. This is
also true for the cultured pearl
industry which now utilizes our
entire commercial harvest for
-------
Freshwater Mollusca
processing into "nuclei" to be
inserted into the pearl "oyster"
for cultured pearl production.
B The body mass is enveloped by the mantle.
Siphons, gills, and foot are major sections
of the body.
1 The mantle functions in respiration and
shell formation.
2 The ventral margins of the mantle are
fused at various places.
a Openings for the foot and byssus.
b Fused to form various siphonal
structures, such as exhalant and
inhalant siphons.
C The foot is a highly elastic organ which
can be extended knife-like into the sub-
strate, expanded at the tip to serve as an
anchor, then the hatchet shaped shell is
forced into the substrate as the foot contracts.
1 Some species are more active burrowers
than others.
2 Although many must have the siphons
reaching the substrate-water interface,
others siphon while burrowed in gravelly
and loose substrates (benthic and
hyporheal).
D Gills are simply modified folds of the
mantle and are an elaborate organ for
respiration, feeding, and often maternal
brooding of eggs and larval stages.
E These animals are considered acephalic,
that is, headless which confused early
scientists trying to describe anterior
or posterior shell features. The head is
at least represented by the mouth. The
siphons are located at the posterior end,
and thus, the animal's anterior end is
directed downward or into the substrate.
Bivalves are the only molluscs without a
radula.
IV The ecology of most of our freshwater
bivalves are poorly known, although nearly
all of our species were described by early
naturalists.
A As in gastropods, the calcium carbonate
shell is a reflection of water chemistry.
For example, the shell can accumulate
large quantities of heavy metals. Where
the periostracum has been worn off or
damaged, the underlying calcium car-
bonate may be severely eroded.
B Dispersal of bivalves has been noticeable.
1 There are a number of the smaller
species of Sphaerndae which are
cosmopolitan or which have been
introduced from both sides of the
Atlantic.
2 An exotic which has been particularly
significant as a nuisance organism is
the "Asian clam, " Corbicula manilensis.
3 At least one species of Unionacea is
circumboreal, Margaritifera
margaritifera
4 The United States has a pronounced
regional Unionid fauna as shown by the
map. The southern Applachians have
a unique fauna. Many species are
already extinct and others are in danger
due to man-induced-changes in the
ecosystem.
C Changes in water quality are difficult to
evaluate in the group since the Umonids
require a fish host, often specific. It
follows that changes in fish fauna will also
be reflected in mussel populations.
1 Mussels have generally been regarded
as indicators of clean water, however,
it has been shown that several species,
including Megalonais gigantea and
Amblema plicata have remnant popu-
lations in streams subjected to a variety
of effluents. Therefore, generalizations
would not be meaningful.
-------
Freshwater Mollusca
2 It should also be obvious that adult
molluscs (remember also that they may
reach upwards of 50 years) which can
remain closed and respire anaerobically
can withstand severe conditions during
limited periods. Reproduction and
maintenance of such populations should
be evaluated.
3 The absence or presence of appropriate
fish hosts must be evaluated. These
factors (1 and 2 above) make mere
presence of adult individuals of mussels
meaningless in water quality investigations
D Food habits need careful study to determine
what elements of the food string is utilized.
Although some sorting of food does take
place on the palps, a variety of phyto and
zooplankton, bacteria, and various sus-
pended and dissolved organic and inorganic
materials pass through the gut, with varying
degrees of utilization.
B Sphaeriids are easily relaxed by placing
them m an acceptable water in a petri
dish and adding a small granule of
pentachlorophenol.
C If naiads are left out on a counter, they
can be "pegged" when the shell gapes.
The peg allows the preservative to reach
the soft parts, and also makes it easier
to insert a scapel to sever the adductor
muscles.
D An unbuffered formalin solution will
effectively decalcify the valves,
destroying a specimen.
VII Final identification to species should be
left to the specialist. Although there are
virtually no undescribed species, the
systematics of this group are unstable.
The main problem involves the matter of
interpretation of taxonomic rules, including
priority.
V Qualitative and quantitative collection of
Unionids or mussels is achieved with difficulty.
The rarer species are often collected by
examining the shell piles accumulated by
foraging muskrats.
A In large streams and lakes scuba diving
techniques using an iron frame for selection
of quadrats yield quantitative results.
B Small streams are difficult to sample.
1 Migratory movements of some mussels
make them relatively inaccessible.
2 Special techniques are needed for
collecting juveniles.
VIII The larger group of bivalves classed as
Unionidae have a variety of names for the
group-including mussels, freshwater clams,
pearl button clams, umonids, pearly naiads,
nayades, najades, etc. The most accepted
group name today is the term Naiads.
Nearly all Naiads have common names which
are specific. These are the same for north
or south, since they evolved during the pearl
button era. They were marketed under such
names as pistol grip, washboard, three
ridge, monkey face, etc.
VI Since anatomical and reproductive features
are useful characters for identification,
bivalves should be relaxed, killed, fixed, and
properly preserved.
A For relaxation, menthol crystals
sprinkled on the water surface of the
water containing them may work.
-------
Freshwater Mollusca
Figure 18. MUSSEL REGIONS OF THE UNITED STATES
(after van der Schalie)
numbers = app. no. of species P - Pacific M - Mississippian
O - Ozarkian c - Cumberlandian N - North Atlantic S - South
Atlantic A - Appalachicolan
REFERENCES
1 Athearn, H.D. How to Find Freshwater
Mollusks in Creek-Size Streams. Amer.
Mala col. Union Bull. 36 31-33. 1970.
2 Isom, Billy G. The Mussel Resource of
the Tennessee River.
3 Morton, J.E. Molluscs. Hutchinson and
Company, Ltd. London. 232 pp. 1958.
4 Parmalee, Paul W. The Freshwater Mussels
of Illinois. Illinois State Museum.
108 pp. 1967.
5 Purchon, R.D. The Biology of the
Mollusca. Pergamon Press. 560 pp.
1968.
6 Russell-Hunter, W. C. A Biology of
Lower Invertebrates. McMillan
Company. 181 pp. 1968.
7 Starrett, William C. A Survey of the
Mussels (Unionacea) of the Illinois
River: A Polluted Stream. Bull.
Illin. Nat. Hist. Surv. 30(5).267-403.
1971.
8 Van der Schalie, Henry and Van der Schalie,
Annette. The Mussels of the
Mississippi River. Amer. Midi. Nat.
44(2) 448-466. 1950.
This outline was prepared by R. M. Sinclair,
Aquatic Biologist, National Training Center,
WPO, EPA, Cincinnati, OH 45268.
-------
FISHES
I INTRODUCTION, What is a fish?
A A fish is a gill-breathing aquatic vertebrate
with fins (exceptions noted).
B Other Aquatic Vertebrates
1 Amphibia - frogs, toads, salamanders
a Modern amphibia do not have scales.
b Tadpole stages easily recognized.
c Pollutional significance not studied
to date. Frogs often observed in
polluted waters but not aquatic
salamanders
2 Reptilia - snakes and turtles
a Relatively independent of water
quality as long as it is not irritating.
b Carnivorous types would be starved
out of polluted areas for lack of food.
3 Mammalia - muskrats, beavers
a Generally inhabit wilderness areas
where heavy pollution is not a
problem.
II STRUCTURE AND PHYSIOLOGY
A Fins
1 A typical fish has two sets of paired
fins, the pectoral and pelvic, compar-
able (homologous) to our arms and legs
respectively. Certain ancient fish
could walk on their lobe-like fins, and
some specialized modern forms like-
wise
2 Unpaired dorsal, anal, and caudal or
tail fins, complete the fin structures
3 Any or all of these may be missing,
and fleshy extra fins such as the
"adipose" fins of trout and salmon,
catfishes, etc., may appear. Extra
paired fins too are known. The dorsal
fin is often divided into two or more
sections known as 1st, 2nd, 3rd, etc. ,
dorsal fins.
4 Fins may be supported by soft rays or
stiff spines or both.
B The body of a typical fish is covered with
scales.
1 Four types of scales are recognized
a The most primitive are bony plates
bearing tooth-like projections as
found in sharks and rays
b Smooth bony plates such as those
of the gar and dogfish are some -
what higher in specialization.
c Thin smooth roundish "cycloid"
scales are characteristic of the
more primitive of the modern
"bony" fishes like herring or trout.
d Roundish scales with tiny spines
or cteni are characteristic of the
highest fishes like the black basses.
These are called ctenoid scales
2 Cycloid and ctenoid scales are non-
living material like hair or fingernails,
covered with a thin layer of living
tissue cells. This tissue is easily
injured as by handling a fish with dry
hands
BI. AQ. 18c. 10.68
-------
Fishes
C Respiration of a typical fish is by means
of blood gills (Cf tracheal gills of insects)
1 Gills, like lungs, are a device for
bringing the blood into close proximity
to the environment.
2 Certain ancient fishes and their modern
descendents, the lungfishes, breathed
air Some of the modern bonyfishes
like certain catfishes also have a
limited air breathing capacity.
D The maintenance of a constant internal
osmotic pressure of their body fluids in
competition with their environment is a
problem which fishes have that terrestrial
animals with their waterproof skins do not
have.
1 The slime covering body and gills is
an important part of the regulatory
mechanism
2 Marine fishes live in an environment
that tends to dehydrate the body Con-
sequently water appears to be swallowed
from time to time in order to replace
that lost through the gills and general
body surface. This may make marine
fishes more susceptible to some toxic
substances which would then be taken
internally instead of simply contacting
the skin externally.
3 Freshwater fishes live in an environ-
ment with an osmotic pressure far
lower than that of body fluids. Toxic
substances would then be less likely
to be taken internally except with food.
Ill REPRODUCTION AND DEVELOPMENT
A Although almost infinite variety exists,
most fishes lay their eggs externally in
the water, at which time the male showers
them with milt Such species are said to
be oviparous. In some species such as
the familiar guppy however, fertilization
is internal and the eggs develop and hatch
in the mother's body. No nourishment
is known to be transmitted from mother
to developing young. Such species are
said to be ovovivipanous. The young
are thus "born alive. "
1 Most fish eggs hatch before all the
food material stored in the egg as
yolk is used up. Embryonic develop-
ment is still going on and they are
truly m a larval or,pre-adult condition.
During their first hours or days of
free life, they are thus still indepen-
dent of their environment for food
This early stage is known as yolk sac
fry or simply sac fry.
2 The young continue to be called fry,
or advanced fry until they approach an
inch in length, when they are referred
to as fingerlings.
3 Fry which differ greatly from the form
of the adult may be referred to as larvae.
B Some fish lay their eggs in the same
general location in which they live as
adults Others travel to some distant
place, such as from lakes or rivers up
into small streams, from deep water to
shoal, from the oce^n to fresh water,
from fresh water to the ocean, etc These
are called breeding migrations.
1 Fish that normally live in freshwater
and travel to the ocean to reproduce
are called catadromous. The fresh-
water eel is the best known example.
2 Those that live in the sea and lay their
eggs in freshwater are called anadro-
mous Striped bass, shad and certain
other herrings, and the salmons are
well known examples. Occasionally,
a group of anadromous fish will get
lost in the inland waters and not be
able to find its way back to the sea.
These are called landlocked varieties
and are usually somewhat smaller than
their non-lan-locked relatives
3 Pollution or other factors whioh either
block a breeding migration or destroy
a spawning bed may completely destroy
-------
Fishes
a species, even though the adults in
their natural habitat are untouched.
IV CLASSIFICATION OF FISHES
Fishes may be classified or grouped in many
.different ways.
A Food, Feeding Habits and Ecological
Interrelationships
Fish, like many other animal groups, in-
clude carnivores, herbivores, and
detritus feeders or scavengers
1 Scavengers may specialize on bottom
feeding like certain suckers, carps,
and catfishes. Others may take any
organic matter they can find, where -
ever they can find it. Scavengers are
often provided with barbels or feeders
which help in locating food, especially
in turbid water.
2 Herbivores may feed on the larger,
vascular plants as some carps or
they may specialize on the microscopic
phytoplankton, in which case they are
called plankton feeders. Plankton
feeders usually have weak mouths and
fine gill rakers for straining the plank
ton out of the water.
3 The carnivorous or predatory species
feed essentially on living animals
They may specialize on invertebrates
or other fish, in which case they may
be called piscivorous.
a Piscivorous fish usually depend
essentially on eyesight for locating
their food, and hence turbid water
is a handicap.
b The carnivorous fish m general
include most of the game fish.
c Small species of fish which are not
used directly by man but are used
extensively as food by piscivorous
species are often referred to as
forage fish.
B Classification with reference to their
desirability or to mode of use by man
has been widely used An example of
such a system is as follows
1 Commercial - those that occur in
sufficient quantities to support a
fishery.
a Food fishes: white-fish, salmon,
cod
b Product fishes sharks, blue back
herrings
2 Game or sport - those captured
essentially for sport. Many species
fall into both this and the commercial
categories such as the trouts, black-
basses, striped bass, etc.
a Gamefish are sometimes considered
to be those which are of interest to
man only for the catching, as the
tarpon.
b Fish which are taken, even though
in sport, but which are also eaten
are then called panfish. Sometimes
panfish refers only to the smaller
of the edible gamefish.
3 Rough fishes are those such as the
gars, and the bowfins, which are of
little or no use to man. Some, such
as the carp, are classed in different
groups in different regions according
to local custom.
C A classification developed with reference
to standard methods of reporting fish
population data for reservoirs is as
follows (Surber '59)
Group 1 Predatory Game Fish - bass,
crappies, trout, etc
Group 2. Non-predatory Game Fish -
sunfish, rock bass, perch, etc.
-------
Fishes
Group 3. Non-predatory Food Fish - carp,
drum, buffalo, suckers, bullheads,
etc.
Group 4 Predatory Food Fish - catfish,
gar, bowfm, etc.
Group 5 Forage Fish (Non-predatory)
gizzard shad, threadfin shad,
Gambusia, minnows, etc.
D The scientific classification system for
fishes lists many thousands of species
Three great groups of living fishes occur
in this country, the Agnatha or jawless
fishes, the Chondrichthyes or cartilaginous
fishes and the Osteichthyes or (modern)
boney fishes. Some additional groups
occur in other parts of the world.
1 The jawless fishes are represented in
fresh water by the lampreys, which
have in recent years invaded the Great
Lakes from the sea and wrecked havoc
on the native species.
2 The cartilaginous fishes are the sharks,
skates, and rays, primarily a marine
group
3 The vast majority of fishes with which
most of us are familiar belong to the
Osteichthyes or bony fishes, a few
typical families are listed below
a The family Acipenseridae or
sturgeons are a primitive group,
famed for their roe which is sold
as caviar. More or less covered
with large, bony plates. Formerly
extremely abundant and large in size
b Family Lepisosteidae - the gars.
These Voracious fish are covered
with hard, enamel-like, rhomboid
scales. Some species may grow
to great size. Widely regarded as
"trash" fish
c The family Salmomdae includes the
trouts, salmons, whitings, and
graylings. Scales are cycloid and
always small, an extra or adipose
fin on the back, eggs are very large,
favored by cold water. In the Pacific
Salmon, but one set of reproductive
cells is formed in the life of the
individual, which therefore dies
after spawning once.
d The family Catostomidae is the
suckers. The head is naked of
scales, jaws toothless, mouth
usually protractile, lips generally
thick and fleshy Feed on plants
and small animals.
e The family Cyprimdae is the carp-
dace-minnow group. Here too the
head is naked, and the body usually
scaled. Ventral fins usually well
back. Teeth are lacking in the jaws.
Certain bones in the back of the
throat known as the pharyngeals are
strongly developed however, and
bear from 1 to 3 series of teeth
which are often of importance in
identification. Upwards of 1800
species, abundant where present
at all, both in numbers and variety
Generally small in size although
Leucosomus corporalis the chub,
roach, or fallfish may reach a
length of 18 inches in the east, and
related species 5 to 6 feet on the
west coast. Because of the many
similar species, this is one of the
most difficult groups in zoology
to identify to species
Two genera, Cyprinus which includes
the common carp, and Carassius
including the goldfish have been
introduced and become widely
established. Both are native to
China Other introduced Cyprimds
have so far not become widely
established.
f Family Ictaluridae, the freshwater
catfishes. Body more or less
elongate, naked Eight barbels or
feelers m head region. Dorsal fin
short, an adipose fin behind. First
ray of dorsal and pectorals developed
as stout spines. Many excellent food
fish. Very tenacious of life.
-------
Fishes
g Family Centrarchidae - sunfishes
and freshwater basses. Scales
ctenoid Dorsal fin continuous
but may be in two sections, the
anterior spined, the posterior rayed
Generally carnivorous. Typical of
eastern North America but have
been widely introduced in other
areas Nest builders.
4 Additional well known families of bony
fishes are listed below.
Polyodontidae
- paddlefishes
Amiidae
- bowfms
Gasterosteidae
- stickelbacks
Cyprinodontidae
- killifishes
Serranidae
- sea basses
Ictaluridae
- freshwater
catfishes
Percidae
- perches and darters
Cottidae
- sculpins
Atherinidae
- silversides
Clupeidae
- herrings
Osmeridae
- smelts
Salmomdae
- whitefishes, trouts,
etc.
Anguiledae
- eel
Poecilndae
- guppies, mosquito-
fishes
Gadidae
- cods, hakes,burbots
Esocidae
- pikes and pickerels
Sciaenidae
- drums
3 Eddy, Samuel. How to Know the Fresh-
water Fishes. Wm C. Brown Co
Dubuque, Iowa 1957
4 Hubbs, C.L., and Lagler, K.F Fishes
of the Great Lakes Region, Bull
Cranbrook Inst. Sci. Bloomfield Hills,
Michigan. 1949.
5 Lagler, K.F. Freshwater Fishery
Biology. Wm. C. Brown Co Dubuque,
Iowa 1952.
6 Surber, E.S. Suggested Standard Methods
of Reporting Fish Population Data for
Reservoirs. Proc. 13th Ann. Conf
S E Assoc Game &. Fish Comm
pp. 313-325 Baltimore, Md
October 25-27, 1959
7 Trautman, M B The Fishes of Ohio
Ohio State Umv Press (An out-
standing example of a state study )
Columbus, Ohio 1957
REFERENCES
1 American Fisheries Society A List of
Common and Scientific Names of Fishes
From the United States and Canada
Special Publication No 2 Am Fish
Soc Dr E.A Seaman, Sec.-Treas
Box 483, McLean, Va (Price $1 00
paper, $2.00 cloth.) 1960
2 Bailey, Reeve M A Revised List of the
Fishes of Iowa with Keys for Identifica-
tion. IN Iowa Fish and Fishing. State
of Iowa Super of Printing (Excel-
lent color pictures ) 1956
This outline was prepared by H.W. Jackson,
Chief Biologist, National Training Center,
Water Programs Operations, EPA, Cincinnati,
OH 45268.
-------
SOME PRIMITIVE FISHES
Class Agnatha, jawless fishes (lampreys and hagfishes) - Family
PETROMYZONTIDAE, the lampreys. Lampetra aepyptera, the
Brook Lamprey A: adult, B; larva (enlarged)
Class Chondrichthyes - cartilagenous fishes (sharks, skates, rays)
Family DASYATIDAE - stingrays. Dasyatis centroura, the Roughtail Stingray
Class Osteichthyes - bony fishes - Family ACIPENSERIDAE, sturgeon.
Acipenser fulvescens, the Lake Sturgeon
Class Osteichthyes - bony fishes - Family POLYODONTIDAE, the
paddlefishes. Polyodon spathula, the Paddlefish. A:side view B:top view
Class Osteichthyes - bony fishes - Family LEPISOSTEIDAE - gars
Lepisosteus osseus, the Longnose Gar
Class Osteichthyes - bony fishes - Family AMIIDAE, bowfins
Aniia calva, the Bowfin
Reproduced with permission; Trautman, 1957.
-------
Family SALMONIDAE
^ Kaffir :
Salmo trutta - brown trout
Salvelinus namaychush - the lake trout
Salmo gairdneri - rainbow (or steelhead) trout
Coregonus clupeaformis - lake whitefish
Salvelinus fontinalis - brook trout Oncorhynchus tshawytscha - the chinook salmon
Reproduced with permission; Trautman, 1947 (except Chinook salmon
after Jordan '05).
-------
Fishes
Family CATOSTOMIDAE - the suckers
Ictiobus cyprinellus - bigmouth buffalofish
Catostomus catostomus - eastern longnose sucker
Hypentelium nigricans - hog sucker
WM
"jj
Moxostoma aureolum - northern shorthead redhorse
Reproduced with permission; Trautman, 1957.
BI.AQ.pl. 9g. 6. 60
-------
Fishes
Family CYPRINIDAE
minnows, carps, and goldfishes
Pimephales promelas - fathead minnow
Notemigonus crysoleucas - golden shiner
Chrosomus erythrogaster - southern redbelly dace
Semotilus atromaculatus - creek chub
Reproduced with permission; Hart, Doudoroff, and Greenbank, 1945.
21-10
-------
Fishes
Family ICTALURIDAE - the freshwater catfishes
Ictalurus nebulosus - brown bullhead
Noturus insignis - margined madtom
Reproduced with permission; Hart, Doudoroff, and Greenbank, 1945.
BI.AQ. pi. 9i. 6.60
-------
Family CENTRARCHIDAE - the sunfishes
Lepomis macrochirus - bluegill
Micropterus salmoides - largemouth bass
Pomoxia nigromaculatus - black crappie
Reproduced with permission; Hart, Doudoroff, and Greenbank, 1945
-------
Fishes
TYPES OF BONY FISHES I
Family GASTEROSTEIDAE, the
Sticklebacks. Eucalia inconstans,
the brook stickleback
Family PERCIDAE - the perches.
Perca flavescens, the yellow perch
Family CYPRINODONTIDAE, the
Killifishes. Fundulus not at us, the
blackstripe topminnow
Family PERCIDAE, the perches.
Etheostoma nigrum, the johnny
darter
Family SERRANIDAE, the sea basses.
Roccus americanus, the white perch
Family COTTIDAE, the sculpins.
Cottus bairdii, the mottled sculpin
Reproduced with permission; Hart, Doudoroff and Greenbank, 1945.
Family CATOSTOMIDAE, the
suckers. Catostomus commersonii,
the white sucker
Family ATHERINIDAE, the silversides.
Labidesthes sicculus, the brook silverside.
BI. AQ. pi. 9m. 6. 60
-------
TYPES OF BONY FISHES
wmmmmm ?
'¦ v. . v1
^v iSar' ¦ ¦ ¦¦¦
Family CLUPEIDAE - herrings
Dorosoma cepedianum - the eastern gizzard shaa
Family POECILIIDAE - livebearers
Gambusia affinis - the mosquitofish
Family ANGUILLEDAE - freshwater eels
Anguilla rostrata - the American eel
"%|sjsig|s
Family ESOCIDAE - pikes
Esox lucius - the northern pike
Reproduced with permission; Trautman, 1957.
BI. AQ. pi. 9m. 6. 60
Family GADDIDAE - codfishes, hakes, haddock, burbot
Lota lota - the eastern burbot
Family SCIAENIDAE - drums
Aplodinotus grunniens - the freshwater drum
i-H
I
H
-------
Fishes
ADIPOSE FINS - in catfishes
Adipose Fin
The Adipose Fin does not extend to the Caudal Fin
(Ictalurus nebulosus)
Adipose Fin
The Adipose Fin extends to the Caudal Fin
(Noturus insignis)
Reproduced with permission; Hart, Doudoroff and Greenbank, 1945.
BI. AQ. pi. 9e. 6. 60
-------
STANDARD LENGTH
LENGTH OF HEAD
SKlOUT
OOffSAL FIN
L
ORIGIN orOOR5AL
LATERAL LINE
PECTORAL FIN
ABOO^EN
liAXt LLARY
PRErlAX \ LLARY
LOWER JAW
ANAL FIN
VENTRAL FIN
OPERCLfc.
5U80PEf?CL£
INTEROPERCLE
PREOPERCLE.
GILL MErieRANES
cauoal Fin
A SOFT- RA YEO nsHt SeriOTJLUS ATKOrtACl/LATUS
Hart, Doudoroff, and
Greenbank, 1945
-------
Giu-MEMBRANCS
G! UL. rtErtB RA NES F~fTEE 0F~ THE /5 Tt-iriUS.
A.\'D SEPARATE
G I LL
MEMBRANCS
Mouth inferior
Gill m E:n b ra nes 5 £ pa r a te a nd connected to
Hart, Doudoroff, and THE 15THM(JS¦
Greenbank, 1945
This outline was prepared by H. W . Jackson
Pharyngeal teeth
Lower pharyngeal bone
Form or pharyngeal teeth /n the
G'RRiNIOAE.
PhaRYMGCALTEETH
Lower phariugcalbonc
Comb-like: teteth/nthe
CATOSTOM/DAE BL AQ. pi. 9c. 6. 60
CO
-------
FIN AND SCALE STRUCTURES
C TENO) 0 5cal£-
Amtctrior
pAf?T
Po5"reF?iof?
PAI7T
A NTCF7I0F?
PART
Posterior
part
A SPINOUS DORSAL HN.
UNDEVELOPED
RAYS
¦ Last i?at split
A SOFT RAYED DORS A L FiH.
5orr ray
CrcLOtD scale:
A DORSAL FIN COMPOSED O F SPINOUS AN O SOFT FfAYS
Reproduced with permission; Hart. Doudoroff, and Greenbank, 1945.
-------
CLASSIFICATION OF FISHES
A classification of fishes and lower vertebrates
following recent researches reported m Romer
1946, Quarterly Review of Biology, Schultz
1948, Ways of Fishes and Berg 1940, Classi-
fication of Fishes. Those preceded by an
asterisk are known from fossil forms only.
Phylum Chordata
Subphylum Hemichordata Acorn worms
Subphylum Tunicata (Urochordata)
Ascidians Sea squirts
(Note the above two groups are not usually
claimed nor studied by vertebrate zoologists).
Subphylum Acrania (without cranium)
Class Cephalochordata (Leptocardi)
Amphioxi
Subphylum Craniata (with cranium) Equals
Vertebrata of many authors
Superclass Agnatha (without jaws, with
bone in part)
Contains four large well differentiated
classes two of which are living and
two known only from fossils. The
latter are the oldest certain remains
of vertebrates and are found in
Ordovician rocks and are lumped
usually under the term Ostracoderms.
Class Petromyzones (Marsipobranchu
or Cyclostomata)
Lampreys. Bone lost.
Class Myxini Hagfishes. Bone lost.
~Class Pteraspides (Heterostraci)
Lower Silurian to upper Devonian
~Class Cephalaspides (Osteostraci)
Upper Silurian to upper Devonian
~Class Euphanenda (Jamoytius)
Superclass Gnathostomata (equals Pisces,
with jaws)
~Class Placodermi (with bone)
Three subclasses (Berg gives them
ranking of class) are usually re-
cognized here. All are fossil.
~Subclass Pterichthyes (Antiarchi)
Middle and upper Devonian.
Pectoral appendages jointed.
~Subclass Coccostei (Arthrodira)
Upper Silurian to upper Devonian
(or to early Mississippian) Jointed
neck, armored fishes.
~Subclass Acanthodii Spiny Sharks.
Upper Silurian to lower Permian
(acme in Lower Devonian).
Class Elasmobranchii (Chondrichthyes)
Sharks and rays.
Bone lost, jaws advanced in structure.
Upper Devonian to recent.
Class Holocephali. Chimaeras, elephant
fishes, and rat fishes (given as sub-
class under Elasmobranchii by some
authors) Cartilaginous, gill cover
developed. Upper Devonian to recent.
Class Osteichthyes (Teleostomi of Berg)
Bony Fishes.
Lower Devonian to recent.
Subclass Choanichthyes
Order Dipnoi Lungfishes Treated
by Berg as a separate class
placed just before Osteichthyes.
Middle (Lower?) Devonian to recent.
Order Crossopterygu Fringed fins.
Lower Devonian to recent.
Latimeria from Southeast Africa
Subclass Actinopterygn Rayed Fins. Middle
Devonian to recent.
Order Chrondrostei Polyptenforms to
Acipenseriformes
Order Holostei Amiiformes, Lepidostei-
formes to Pholiodophoriformes
Order Teleostei, Clupeiformes, etc.
-------
Classification of Fishes
FIRST DORSAL FIN
SECOND DORSAL FIN
NOSTRIL
CAUDAL FIN
(UPPER LOBE)
^^CLASPER
VENTRAL FIN
CAUDAL FIN
(LOWER LOBE)
Figure 1. A TYPICAL SHARK SHOWING THE IMPORTANT EXTERNAL PARTS
EYE
SPIRACLE
PECTORAL FIN
VENTRAL FIN
CLASPER
OOftSAL FIN
SPINE
TAIL
Figure 2. A TYPICAL RAY SHOWING THE IMPORTANT EXTERNAL PARTS
-------
Classification of Fishes
FIRST DORSAL FIN
LATERAL LINE
SECOND DORSAL FIN
PREORBITAL
SNOUT
CAUDAL FIN
CAUDAL PEOUNCLE
LOWER JAW
SILL OPENING
VENTRAL FIN
PECTORAL FIN
Figure 3. A TYPICAL BONY FISH SHOWING THE IMPORTANT EXTERNAL PARTS
DORSAL FIN LATERAL LINE
UPPER JAW
LOWER JAW
CAUDAL PEOUNCLE
ANAL FIN
VENTRAL FIN
PECTORAL FIN
Figure 4. A FLATFISH, AN ABERRANT FORM OF BONY FISH
-------
SECTION D
BACTERIA AND FUNGI: REDUCERS
This group of organisms as a group and as individuals exhibit a remarkable
catholicity of taste. There is literally no known organic constituent of living
things that cannot be used as food by at least one kind of bacterium or fungus.
Clearly, this is essential to the continuing flow of carbon. Without this
process carbon find energy would slowly but certainly accumulate in unused
products of plant or animal metabolism. As will be shown in the following
three outlines, the metabolsim of this group is marked by an enormous
complexity and diversity. Underneath this complexity there is a fundamental
oneness of mechanism shared by all living organisms. This unity in metab-
olism is further marked by the completing role of the reducers in nature as
the indispensable feeders of the furnace of photosynthesis.
Contents of Section D
Outline No.
Biological Reducers (The Role of Fungi in the
Biodegradation of Organic Matter)
23
Bacteriological Indicators of Water Pollution
24
Fungi and the "Sewage Fungus" Community
-------
BIOLOGICA L REDUCERS
(The Role of Fungi in the Biodegradation of Organic Matter)
X INTRODUCTION
A Biological Classification System Based
on Nutrition
Based on their fundamental nutritional
mechanisms, all organisms can be
segregated mto three major groups, or
"kingdoms" instead of the traditional two
("plant" vs "animal") kingdoms.
These are
1 The green plants
The nutrition of the green plants is
based on photosynthesis in which
sunlight, carbon dioxide, water, and
the chlorophyll of the green plant
enter into a reaction (or series of
reactions) resulting in the synthesis of
sugars. From these fundamental
building blocks the green plant synthe-
sizes all the other organic compounds
(utilizing other elements, notably
nitrogen, phosphorus, sulfur, calcium,
potassium, iron, and other elements
in trace amounts).
2 The animals
Animals obtain their organic com-
pounds, required for nutrition and
growth, by ingestion of solid foods
into the body structure. This food
may be the bodies of green plants,
the bodies of other animals, or it
may be products, secretions, or
excretions of either form of life.
3 The fungi
In a broad sense this includes both the
filamentous fungi and the bacteria.
These organisms obtain their food in
solution, that is, they secrete extra-
cellular enzymes into their environment,
which break down complex organic
matter sufficiently that it diffuses
through the selectively permeable
cell walls and cell membranes of
the organism. Once within the cell
structure, these dissolved organic
materials are used for nutrition and
growth in a manner comparable with
that of the animals.
B Scope of this presentation
This discussion is concerned with the
role of the fungi (including bacteria) in
the natural self-purification processes
occurring in the environment.
II NUTRITIONAL PROPERTIES OF THE
FUNGI
A From species to species the fungi vary
enormously in the details of their
nutritional processes, and therefore,
in the substances they are capable of
using for food. In addition, they differ
greatly from species to species in their
environmental "preferences", resulting
in a wide variety of species complexes
which can be found from one aquatic
habitat to another. Among the factors
having great influence in selection of the
species and species complexes which
will appear in a given habitat may be
cited
1 Oxygen, resulting in selectivity for
strictly aerobic forms, or facultative
forms.
2 pH, resulting in a tendency to select
the forms having optimum growth
and survival properties within a
limited range of pH values. For
example, most bacteria tend to grow
best in pH levels at or near neutrality,
while the filamentous fungi commonly
have optimum growth properties
WP. NAP. 23.4. 73
-------
Biological Reducers
somewhat on the acid side of
neutrality, say pH 5 to pH 7, or
sometimes even lower.
3 Presence of certain organic growth
requirements, some kinds of bacteria
require external sources for complex
organic substances that they require
but cannot synthesize for themselves.
Such organisms are, of course, highly
limited and highly specific in the
habitats in which they can compete
successfully for the food.
4 Nature of the substrate (food) available
to the organisms, some kinds of
bacteria are highly effective in utilizing
carbohydrates (members of the
Enterobactericeae, for example).
There is a great deal of specificity
from one kind of organism to another
in the carbohydrates which can be
metabolized. Others are not particularly
active in using carbohydrates, but
apparently are highly successful in
using other forms of organic matter
for food. (Pseumonadaceae, for
example, are quite versatile in their
nutritional capabilities.
B Cycles of Carbon, Nitrogen,
Phosphorus
and
Incidental to their nutrition on which
organic substances are used for food,
the fungi play a vital role in the return
to a mineral state of elements bound up
in organic substances. The classical
cycles of carbon, nitrogen, and phos-
phorus illustrate this role. These
cycles may be represented diagramatically
in a variety of degrees of complexity.
The accompanying diagrams are intended
to show the essential linkage of the fungi
in completing the cycling of these sub-
stances, essential to life, which
eventually would be bound up in the bodies
of plants, animals, and their products,
in the absence of a mechanism for
returning the substances to a mineral
state, with renewed availability to the
green plant.
Resynthesis
of animal
carbohydrates
and fats
Plant
synthesis of
sugars, starches,
cellulose and
other carbo-
hydrates, fats
and oils
&
Carbon
dioxide
CARBON CYCLE (adapted after Hilliard, 1945)
-------
Biological Reducers
—^"Tprial Reduction)
St73atte
NO
NITROGEN CYCLE (after Allen, 1938)
-------
Biological Reducers
.VOLCANOES
mine drainage
BACTERIA
bacteria 40
A NIMAU PROTEIN
PLANT PROTEIN
SULFUR CYCLE (after Salle, 1948)
. Animals -»• Man
Carcasses Pi
Manure (bone)
>lants
\ I
1
\
Birds
Sewage Guano
Soil and freshwater environment
Fish
Oceans IShell-
fish
/
Deep Sea
Deposits
PHOSPHORUS CYCLE (after Salle, 1948)
-------
Biological Reducers
C Fate of Organic Matter Used for Nutrition
The complex and interrelated cycles
illustrated above can be simplified along
the following lines-
(plants)
PRODUCERS ¦
(animals)
-*• CONSUMERS
REDUCERS
In the process of reducing organic matter
the fungi are playing the one "role" that
appears basic with all forms of life
to stay alive, to grow and reproduce.
The utilization of the organic matter,
then, does not result purely in the direct
production of inorganic mineral substances.
Much of this organic matter is utilized
in the development of new cells of the
bacteria and filamentous fungi.
This may be represented diagrammatically
according to the figure showing sequence
of metabolic change during oxidation of
carbonaceous material. The figure
represents "favorable conditions".
As environmental conditions become
"less favorable" the reserves of unused
food persist for a greater period of time
but the ultimate conversion to cell sub-
stance or to mineralized decomposition
products ultimately will be the same.
z
UJ
2
D
O
-J
<
0
5
UJ
1
o
tr
o
z
UJ
o
X
o
(4) UN UsEDEXOTIC FO00
SUBSTANCE
ADSORBE
MATERIAL
• UNUSED
;Rt ADILY
! AvAILABl
: FOOD
(I) OXYGEN USEO
(Moterial destroyed by biochemical oxidation)
4 5
TIME UNITS
SEQUENCE OF METABOLIC CHANGE DURING OXIDATION
OF CARBONACEOUS MATERIAL
(Favorable Conditions)
(Ettinger, l'» *>1
-------
Biological Reducers
III ENVIRONMENTA L INTERRELATIONSHIPS
A Influence of Fungi on Their Environment
1 Dissolved oxygen
This topic is discussed at length
elsewhere in this course manual.
It is sufficient here to recall that the
biochemical process of respiration,
going on at all times in all living
cells, results in utilization o'f oxygen
(when available) as a hydrogen acceptor
in the final stages of the respiratory
reactions. When abundant food is
present, and extremely large popu-
lations of bacteria develop rapidly,
the oxygen in the aquatic habitat
sometimes is used up more rapidly
than it can be replenished from the
air or from other sources. In such
cases, extremes of oxygen depletion
may occur, even including anaerobic
situations.
2 pH
Asa result of utilization of car-
bohydrates, organic acids with pH
lower than neutrality (pH 7) may
occur, with the effect of restricting
the value of the habitat for some life
forms, while tending to favor others
(such as the filamentous fungi).
3 Mineralization
Return of sulfates, nitrates, phos-
phorus, and other elements to the
aquatic environment as a result of
bacterial activity has a "fertilizing"
effect on such habitats. Often the
result is prolific "blooms" of algae
which in themselves may create a
nuisance and may in turn bring about
widely fluctuating levels of dissolved
oxygen on a diurnal basis.
4 Antibiotic effects
Some kinds of bacteria and filamentous
fungi excrete substances having
antibiotic effects, killing or inactivating
certain other forms of life. The
Actinomyces, certain members of
the genus Pseudom'onas and certain
forms of the filamentous fungus
Penicillium are notable examples of
antibiotic-producing members of the
fungi.
B Influence of the Environment on Fungi
Environmental factors critical in
selection of populations of bacteria and
filamentous fungi which may be expected
in an aquatic environment include, but
are not necessarily limited to-
1 pH
2 Temperature
3 Salinity
4 Other organisms, especially predators
or antibiotic-producers
5 Available nutrient materials and
special growth factors if required
IV POLLUTION CONTROL SIGNIFICANCE
A In the aquatic environment the bacteria
and filamentous fungi utilize organic
substrates, thereby carrying on functions
regarded desirable to humans and
generally lumped as "natural self
purification". When the amount of
organic matter in the environmental
water is not excessive and properties
of the water are suitable for growth of
these organisms, mineralization goes
on without sharp buildup of carbon,
nitrogen, phosphorus, and other elements.
Further, oxygen required in respiratory
processes does not significantly deplete
the level dissolved in the water, and is
readily replaced m solution from the
overlying air. Nuisance conditions do
not develop here. On the other hand,
in situations whdre the concentration of
organic matter is great, another set of
conditions can develop with excessive
buildup of minerals in solution, depletion
of oxygen, and violent changes in the
biota capable of surviving in such habitats.
-------
Biological Reducers
B In wastewater treatment plants, engineers
make use of the properties of the fungi in
returning organic matter to circulation
through development of structures and
processes designed to utilize the capa-
bilities of these organisms under
controlled conditions in order to minimize
the amount of organic material discharged
to the aquatic habitat. Trickling filters,
activated sludge treatment systems,
sludge digestion systems, and septic
tanks with associated tile fields all make
use of biological processes for such
processes. Bacterial processes have
been regarded as predominant in such
systems. Increasing evidence is
appearing that filamentous fungi have a
significant role to play, particularly in
trickling filters.
REFERENCES
Any one of the many basic text books
available on bacteriology.
1 Ettinger, M.B. Biochemical Oxidation
Characteristics of Stream-Pollutant
Organics. Industrial and
Engineering Chemistry, 48 256.
February 1956.
2 Kabler, P.W. Selection and Adaptation
of Microorganisms in Waste
Treatment. American Journal of
Public Health. 50 215. February
1960.
This outline was prepared by Harold L.
Jeter, Director, National Training Center,
WPO, Cincinnati, OH 45268.
-------
BACTERIOLOGICAL INDICATORS OF WATER POLLUTION
Part 1. General Concepts
I INTRODUCTION
A Bacterial Indication of Pollution
1 In the broadest sense, a bacterial
indicator of pollution is any organism
which, by its presence, would demon-
strate that pollution has occurred, and
often suggest the source of the pollution.
2 In a more restrictive sense, bacterial
indicators of pollution are associated
primarily with demonstration of con-
tamination of water, originating from
excreta of warm-blooded animals
(including man, domestic and wild
animals, and birds).
B Implications of Pollution of Intestinal
Origin
1 Intestinal wastes from warm-blooded
animals regularly include a wide
variety of genera and species of
bacteria. Among these the coliform
group may be listed, and species of
the genera Streptococcus, Lactobacillus,
Staphylococcus, Proteus, Pseudomonas,
certain spore-forming bacteria, and
others.
2 In addition, many kinds of pathogenic
bacteria and other microorganisms
may be released in wastes on an inter-
mittent basis, varying with the geo-
graphic area, state of community
health, nature and degree of waste
treatment, and other factors. These
may include the following
a Bacteria Species of Salmonella,
Shigella, Leptospira, Brucella,
Mycobacterium, and Vibrio comma.
b Viruses. A wide variety, including
that of infectious hepatitis, Polio -
viruses, Coxsackie virus, ECHO
viruses (enteric cytopathogenic
human orphan -- "viruses in search
of a disease"), and unspecified
viruses postulated to account for
outbreaks of diarrheal and upper
respiratory diseases of unknown
etiology, apparently infective by
the water-borne route.
c Protozoa: Endamoeba histolytica
3 As routinely practiced, bacterial
evidence of water pollution is a test
for the presence and numbers of
bacteria in wastes which, by their
presence, indicate that intestinal
pollution has occurred. In this con-
text, indicator groups discussed in
subsequent parts of this outline are
as follows-
a Coliform group and certain sub-
groupings
b Fecal streptococci and certain
sub groupings
c Miscellaneous indicators of pollution
4 Evidence of water contamination by
intestinal wastes of warm-blooded
animals is regarded as evidence of
health hazard in the water being tested.
II PROPERTIES OF AN IDEAL INDICATOR
OF POLLUTION
A An "ideal" bacterial indicator of pollution
should-
1 Be applicable m all types of water
W. BA. 48g. 12.72
-------
Bacteriological Indicators of Water Pollution
2 Always be present in water when
pathogenic bacterial constituents of
fecal contamination are present.
Ramifications of this include --
a Its density should have some direct
relationship to the degree of fecal
pollution.
b It should have greater survival time
in water than enteric pathogens,
throughout its course of natural
disappearance from the water body,
c It should disappear rapidly from
water following the disappearance
of pathogens, either through natural
or man-made processes,
d It always should be absent in a
bacteriologically safe water.
3 Tests for compliance with established
standards in cases involving the pro-
tection or prosecution of municipalities,
industries, etc.
B Treatment Plant Process Control
1 Water treatment plants
2 Wastewater treatment plants
C Water Quality Surveys
1 Determination of intestinal pollution
in surface water to determine type and
extent of treatment required for com-
pliance with standards
2 Tracing sources of pollution
3 Lend itself to routine quantitative
testing procedures without interference
or confusion of results due to extra-
neous bacteria
4 Be harmless to man and other animals
B In all probability, an "ideal" bacterial
indicator does not exist. The discussion
of bacterial indicators of pollution in the
following parts of this outline include
consideration of the merits and limitations
of each group, with their applications in
evaluating bacterial quality of water.
IE APPLICATIONS OF TESTS FOR
POLLUTION INDICATORS
A Tests for Compliance with Bacterial
Water Quality Standards
1 Potability tests on drinking water to
meet Interstate Quarantine or other
standards of regulatory agencies.
2 Determination of bacterial quality of
environmental water for which quality
standards may exist, such as shellfish
waters, recreational waters, water
resources for municipal or other
supplies.
3 Determination of effects on bacterial
flora, due to addition of organic or
other wastes
D Special Studies, such as
1 Tracing sources of intestinal pathogens
in epidemiological investigations
2 Investigations of problems due to the
Sphaerotilus group
3 Investigations of bacterial interference
to certain industrial processes, with
respect to such organisms as Pseudo-
monas, Achromobacter, or others
IV SANITARY SURVEY
The laboratory bacteriologist is not alone in
evaluation of indication of water pollution of
intestinal origin. On-site study (Sanitary
Survey) of the aquatic environment and
adjacent areas, by a qualified person, is a
necessary collateral study with the laboratory
work and frequently will reveal information
regarding potential bacteriological hazard
which may or may not be demonstrated
through laboratory findings from a single
sample or short series of samples.
-------
Bacteriological Indicators of Water Pollution
Part 2. The Coliform Group and Its Constituents
I ORIGINS AND DEFINITION
A Background
1 In 1885, Escherich, a pioneer bacteri-
ologist, recovered certain bacteria from
human feces, which he found in such
numbers and consistency as to lead him
to term these organisms "the charac-
teristic organism of human feces. "
He named these organisms Bacterium
coli-commune and _B. lactis aerogenes.
In 1895, another bacteriologist,
Migula, renamed B. coli commune as
Escherichia coli, which today is the
official name for the type species.
2 Later work has substantiated much of
the original concept of Escherich, but
has shown that the above species are
in fact a heterogeneous complex of
bacterial species and species variants.
a This heterogeneous group occurs not
only in human feces but representatives
also are to be found in many environ-
mental media, including sewage,
surface freshwaters of all categories,
in and on soils, vegetation, etc.
b The group may be subdivided into
various categories on the basis of
numerous biochemical and other
differential tests that may be applied
B Composition of the Coliform Group
1 Current definition
As defined in "Standard Methods for the
Examination of Water and Wastewater"
(l3th ed): "The coliform group includes
all of the aerobic and facultative
anaerobic, Gram-negative, nonspore-
forming rod-shaped bacteria which
ferment lactose with gas formation
within 48 hours at 35°C "
2 The term "coliforms" or "coliform
group" is an inclusive one, including
the following bacteria which may
meet the definition above
a Escherichia coli, E. aurescens,
E. freundu, E. intermedia
b Aerobacter aerogenes, A. cloacae
c Biochemical intermediates between
the genera Escherichia and Aero-
bacter
£ The above terminology is in accordance
with the current editions of Standard
Methods and Bergey's Manual of Deter-
minative Bacteriology and will be
consistent throughout this manual until
these sources are modified.
3 There is no provision m the definition
of coliform bacteria for "atypical" or
"aberrant" coliform strains,
a An individual strain of any of the above
species may fail to meet one of the
criteria of the coliform group,
b Such an organism, by definition, is
not a member of the coliform group,
even though a taxonomic bacteriologist
may be perfectly correct in classifying
the strain in one of the above species.
H SUBDIVISION OF COLIFORMS INTO
"FECAL" AND "NONFECAL"
CATEGORIES
A Need
Single-test differentiations between
coliforms of "fecal" origin and those of
"nonfecal" origin are based on the
assumption that typical E. coli and
closely related strains are of fecal
origin while A. aerogenes and its close
relatives are not of direct fecal origin.
(The latter assumption is not fully borne
out by investigations at this Center.
See Table 1, IMViC Type --++). A
number of single differential tests have
been proposed to differentiate between
"fecal" and "nonfecal" coliforms.
-------
Etactenological Indicators of Water Pollution
Without discussion of their relative merits,
several may be cited here-
B Types of Single-Test Differentiation
1 Determination of gas ratio
Fermentation of glucose by E. coli
results in gas production, with
hydrogen and carbon dioxide being
produced in equal amounts.
Fermentation of glucose by A.
aero genes results in generation of
twice as much carbon dioxide as
hydrogen.
Further studies suggested absolute
correlation between H^/CO^ ratios
and the terminal pH resulting from
glucose fermentation. This led to the
substitution of the methyl red test.
2 Methyl red test
Glucose fermentation by E. coli
typically results in a culture medium
having terminal pH m the range 4.2-
4. 6 (red color a positive test with the
addition of methyl red indicator).
A. aerogenes typically results in a
culture medium having pH 5. 6 or
greater (yellow color, a negative test).
3 Indole
When tryptophane, an amino acid, is
incorporated in a nutrient broth,
typical E. coli strains are capable of
producing indole (positive test) among
the end products, whereas A. aerogenes
does not (negative test).
In reviewing technical literature, the
worker should be alert to the method
used to detect indole formation, as the
results may be greatly influenced by
the analytical procedure.
4 Voges-Proskauer test (acetylmethyl
carbinol test)
The test is for detection of acetylmethyl
carbinol, a derivative of 2, 3, butylene-
glycol, as a result of glucose
fermentation in the presence of
peptone. A. aerogenes produces
this end product (positive test)
whereas E. coli gives a negative test.
a Experience with coliform cultures
giving a positive test has shown a
loss of this ability with storage on
laboratory media for 6 months to
2\years, in 20 - 25% of cultures
(105 out of 458 cultures).
b Some workers consider that all
coliform bacteria produce acetyl-
methyl carbinol in glucose metab-
olism. These workers regard
acetylmethyl carbinol-negative
cultures as those which have
enzyme systems capable of further
degradation of acetylmethyl
carbinol to other end products
which do not give a positive test
with the analytical procedure.
Cultures giving a positive test for
acetylmethyl carbinol lack this
enzyme system.
c This reasoning leads to a hypothesis
(not experimentally proven) that the
change of reaction noted in certain
cultures in 4.a above is due to th'6
activation of a latent enzyme system.
5 Citrate utilization
Cultures of E. coli are unable to use
the carbon of citrates (negative test)
in their metabolism, whereas cultures
of_A. aerogenes are capable of using
the carbon of citrates m their metab-
olism (positive test).
Some workers (using Simmons Citrate
Agar) incorporate a pH indicator
(brom thymol blue) in'the culture
medium in order to demonstrate the
typical alkaline reaction (pH 8.4 -9.0)
resulting with citrate utilization.
6 Elevated temperature (Eijkman) test
a The test is based on evidence that
E. coli and other coliforms of fecal
-------
Bacteriological Indicators of Water Pollution
origin are capable of growing and
fermenting carbohydrates {glucose
or lactose) at temperatures signif-
icantly higher than the body tem-
perature of warm-blooded animals.
Organisms not associated with direct
fecal origin would give a negative
test result, through their inability
to grow at the elevated temperature.
b While many media and techniques
have been proposed, EC Broth, a
medium developed by Perry and
Hajna, used as a confirmatory
medium for 24 hours at 44. 5 ±
0.2oc are the current recommended
medium and method of choice.
While the "EC" terminology of the
medium suggests "j3. coli" the
worker should not regard this as a
specific procedure for isolation of
_E. coll.
c A similar medium. Boric Acid
Lactose Broth, has been developed
by Levme and his associates This
medium gives results virtually
identical with those obtained from
EC Broth, but requires 48 hours of
incubation.
d Elevated temperature tests require
incubation in a water bath Standard
Methods 13th Ed. requires this
temperature to be 44.5 + 0.2°C.
Various workers have urged use of
temperatures ranging between
43.0°C and 46. 0°C. Most of these
recommendations have provided a
tolerance of + 0. 5° C from the rec-
ommended levels. However, some
workers, notably in the Shellfish
Program of the Public Health Service,
stipulate a temperature of 44. 5 +
0. 2° C. This requires use of a water
bath with forced circulation to main-
tain this close tolerance. This
tolerance range has been instituted
in the 13th Edition of Standard Methods
and the laboratory worker should
conform to these new limits.
e The reliability of elevated temper-
ature tests is influenced by the
time required for the newly-
inoculated cultures to reach the
designated incubation temperature.
Critical workers insist on place-
ment of the cultures in the water
bath within 30 minutes, at most,
after inoculation
7 Other tests
Numerous other tests for differentiation
between coliforms of fecal vs. nonfecal
origin have been proposed. Current
studies suggest little promise for the
following tests in this application
uric acid test, cellobiose fermentation,
gelatin liquefaction, production of
hydrogen sulfide, sucrose fermentation,
and others.
C IMViC Classification
1 In 1938, Parr reported on a review of
a literature survey on biochemical tests
used to differentiate between coliforms
of fecal vs. nonfecal origin. A summary
follows
No. of times
Test used for dif-
ferentiation
Voges-Proskauer 22
reaction
Methyl red test 2 0
Citrate utilization 20
Indole test 15
Uric acid test 6
Cellobiose fermentation 4
Gelatin liquefaction 3
Eijkman test 2
Hydrogen sulfide 1
production
Sucrose fermentation 1
a-Methyl-d-glucoside 1
fermentation
-------
Bacteriological Indicators of Water Pollution
2 Based on this summary and on his own
studies, Parr recommended utilization
of a combination of tests, the indole,
methyl red, Voges-Proskauer, and the
citrate utilization tests for this differ-
entiation. This series of reactions is
designated by the mnemonic "IMViC".
Using this scheme, any coliform culture
can be described by an "IMViC Code"
according to the reactions for each
culture. Thus, a typical culture of
E. coli would have a code ++--, and a
typical A. aerogenes culture would
have a code --++.
3 Groupings of coliforms into fecal,
non-fecal, and intermediate groups,
as shown in "Standard Methods for the
Examination of Water and Wastewater"
are shown at the bottom of this page.
D Need for Study of Multiple Cultures
All the systems used for differentiation
between coliforms of fecal vs. those of
nonfecal origin require isolation and study
of numerous pure cultures. Many workers
prefer to study at least 100 cultures from
any environmental source before attempting
to categorize the probable source of the
coliforms.
Ill NATURAL DISTRIBUTION OF COLIFORM
BACTERIA
A Sources of Background Information
Details of the voluminous background of
technical information on coliform bacteria
recovered from one or more environ-
mental media are beyond the scope of
this discussion. References of this
outline are suggested routes of entry
for workers seeking to explore this
topic.
B Studies on Coliform Distribution
1 Since 1960 numerous workers
have engaged in a continuing study of
the natural distribution of coliform
bacteria and an evaluation of pro-
cedures for differentiation between
coliforms of fecal vs. probable non-
fecal origin. Results of this work
have special significance because:
a Rigid uniformity of laboratory
methods have been applied through-
out the series of studies
b Studies are based on massive
numbers of cultures, far beyond
any similar studies heretofore'
reported
Groupings of Coliforms into Fecal, Nonfecal and Intermediate Groups
Organism
Indole
Methyl
red
Voges-
Proskauer
Citrate
E. coli, Variety I
Variety II
+
+
+
-
-
E. freundii
(Intermediates)
Variety I
-
+
-
±
Variety II
+
+
-
+
A. aerogenes
Variety I
-
-
+
+
Variety II
+
-
+
+
-------
Bacteriological Indicators of Water Pollution
c A wider variety of environmental and
biological sources is being studied
than in any previous series of reports.
d All studies are based on freshly
recovered pure culture isolates
from the designated sources.
e All studies are based on cultures
recovered from the widest feasible
geographic range, collected at all
seasons of the year. It is believed
that no more representative series
of studies has been made or is in
progress.
2 Distribution of coliform types
Table 1 shows the consolidated results
of coliform distributions from various
biological and environmental sources.
a The results of these studies show a
high order of correlation between
known or probable fecal origin and
the typical El. coli IMViC code
{++—). On the other hand,
human feces also includes
numbers of A. aerogenes and other
IMViC types, which some regard as
"nonfecal" segments of the coliform
group. (Figure 1)
b The majority of coliforms attributable
to excretal origin tend to be limited
to a relatively small number of the
possible IMViC codes, on the other
hand, coliform bacteria recovered
from undisturbed soil, vegetation,
and insect life represent a wider
range of IMViC codes than fecal
sources, without clear dominance of
any one type. (Figure 2)
c The most prominant IMViC code
from nonfecal sources is the inter-
mediate type, -+-+, which accounts
for almost half the coliform cultures
recovered from soils, and a high
percentage of those recovered from
vegetation and from msects. It
would appear that if any coliform
segment could be termed a "soil
type" it would be IMViC code
d It should not be surprising that
cultures of typical E_. coli are
recovered in relatively smaller
numbers from sources judged,
on the basis of sanitary survey,
to be unpolluted. There is no
known way to exclude the influence
of limited fecal pollution from small
animals and birds in such environ-
ments.
e The distribution of coliform types
from human sources should be
regarded as a representative value
for large numbers of sources
Investigations have shown that there
can be large differences in the
distribution of IMViC types from
person to person, or even from an
individual.
3 Differentiation between coliforms of
fecal vs. nonfecal origin
Table 2 is a summary of findings
based on a number of different criteria
for differentiating between coliforms
of fecal origm and those from other
sources.
a IMViC type +H— is a measurement
of E_. coli. Variety I, and appears
to give reasonably good correlation
between known or highly probable
fecal origin and doubtful fecal origin
b The combination of IMViC types,
++--, +—t and -+--, gives
improved identification of probable
fecal origin, and appears also to
exclude most of the coliforms not
found in excreta of warm-blooded
animals in large numbers
c While the indole, methyl red,
Voges Proskauer, and citrate
utilization tests, each used alone,
appear to give useful answers when
applied only to samples of known
pollution from fecal sources, the
interpretation is not as clear when
applied to coliforms from sources
believed to be remote from direct
fecal pollution.
-------
Table 1. COLIFORM DISTRIBUTION BY IMViC TYPES AND ELEVATED TEMPERATURE
TEST FROM ENVIRONMENTAL AND BIOLOGICAL SOURCES
IMViC
type
Vegetation
Insects
Soil
Fecal sources
Poultry
Undisturbed
Polluted
Hurr
lan
Lives
.tock
iMa.
stir a ins
% of
total
No.
strains
% of
total
No.
strains
% of
total
No.
strains
% of
total
No.
strains
% of
total
No.
strains
% of
total
No.
strains
% of
total
i
«H™-
128
10. 6
134
12. 4
131
5. 6
536
80. 6
3932
87. 2
2237
95. 6
1857
97. 9
--++
237
19. 7
113
10. 4
443
18. 8
13
2. 0
245
5. 4
0
<0. 1
1
0. 1
23
1. 9
0
<0. 1
78
3. 3
1
0. 2
99
2. 2
14
0. 6
20
1.1
+++-
2
0. 2
0
<0. 1
7
0. 3
0
<0. 1
106
2. 4
59
2. 5
o
<0. 1
-+- +
168
14. 0
332
30. 6
1131
48. 1
87
13. 0
50
1.1
1
<0. 1
5
0. 3
++- +
116
9. 6
118
10. 9
87
3. 7
22
3. 3
35
0. 8
27
1. 2
11
0. 6
-+++
32
2. 7
28
2. 6
181
7. 7
5
0. 7
21
0. 5
0
<0. 1
0
<0. 1
++++
291
24. 2
254
23. 4
159
6. 8
0
<0 1
6
0. 1
0
<0. 1
0
<0.1
+-++
88
7. 3
46
4. 2
67
2. 9
0
<0. 1
14
0. 2
0
<0. 1
0
<0. 1
+
87
7. 2
42
3. 9
4
0. 2
1
0. 2
2
<0. 1
0
<0. 1
0
<0. 1
-++-
5
0. 4
0
<0. 1
1
<0. 1
0
<0. 1
0
<0. 1
0
<0. 1
0
<0. 1
19
1. 6
0
<0. 1
53
2. 3
0
<0. 1
0
<0. 1
0
<0. 1
0
<0. 1
+-+-
2
0. 2
0
<0. 1
6
0. 3
0
<0. 1
0
<0. 1
0
<0. 1
0
<0. 1
+—h
5
0. 4
8
0. 7
0
<0. 1
0
<0. 1
0
<0. 1
0
<0. 1
0
<0. 1
+
0
<0. 1
9
0. 8
0
<0. 1
0
<0. 1
2
<0. 1
0
<0. 1
2
<0. 1
Total
1203
1084
2348
•
665
4512
2339
1896
No. EC +
169*
162*
216
551
4349
2309
1765
% EC +
14. 1*
14. 9*
9. 2
82. 9
96. 4
98. 7
93. 0
*120 of these *129 of these
were were
15 —H+, 2 7 —I—(-,
-------
Bacteriological Indicators of Water Pollution
HUMAN
EC ® 96 4
BALB ® 94 7
SUMMARY
Type
Percent posifive
t + —
91.8
-+—
1.5
f
0.1
2 8
EC ©
96 3
BAIB
95 3
- - - 4 - + - +
LIVESTOCK
EC ® 98 7
BALB ® 98 6
POULTRY
EC ® 93 0
BALB ® 92.5
GpLIFORMS
67 Soil Samples
(Geltlieicli. el .il.)
+ ^ l
EH3
Undistu rbed
Soil
Polluted
Soil
-------
Bacteriological Indicators of Water Pollution
Table 2. COMPARISON OF COLIFORM STRAINS ISOLATED FROM WARM-BLOODED ANIMAL
FECES, FROM UNPOLLUTED SOILS AND POLLUTED SOILS WITH USE OF THE
IMViC REACTIONS AND THE ELEVATED TEMPERATURE TEST IN EC MEDIUM
AT 44.50 C (+0.5°) (12th ed. 1965, Standard Methods for the Examination of Water
and Wastewater)
Test
Warm-blooded
animal feces
Soil.
Unpolluted
Soil'
Polluted
Vege-
tation
Insects
+ + - -
91. 8%
5. 6%
80. 6%
10. 6%
12. 4%
+ and
- + —
93. 3%
8. 9%
80. 7%
12, 5%
13. 2%
Indole positive
94. 0%
19. 4%
82. 7%
52. 5%
52. 4%
Methyl red positive
96. 9%
75. 6%
97. 9%
63. 6%
79. 9%
Voges-Proskauer positive
5. 1%
40. 7%
97. 3%
56. 3%
40. 6%
Citrate utilizers
3. 6%
88. 2%
19. 2%
85. 1%
86. 7%
Elevated temperature (EC),
positive
96. 4%
9. 2%
82. 9%
14. 1%
14. 9%
Number of cultures
studied
8, 747
2, 348
665
1, 203
1,084
Total Pure Cultures Studied 14,047
d The elevated temperature test gives
excellent correlation with samples
of known or highly probable fecal
origin. The presence of smaller,
but demonstrable, percentages of
such organisms in environmental
sources not interpreted as being
polluted could be attributed largely
to the warm-blooded wildlife 111 the
area, including birds, rodents, and
other small mammals.
e The elevated temperature test yields
results equal to those obtained from
the total IMViC code. It has marked
advantages in speed, ease and
simplicity of performance, and yields
quantitative results for each water
sample. Therefore, it is regarded
as the method of choice for differ-
entiation between coliforms of
probable direct fecal origin and those
which may have become established
in the bacterial flora of the aquatic
or terrestrial habitat.
IV EVALUATION OF COLIFORMS AS
POLLUTION INDICATORS
A The Coliform Group as a Whole
1 Merits
a The absence of coliform bacteria is1
evidence of a bactenologically safe
water.
b The density of coliforms is roughly
proportional to the amount of
excretal pollution present.
c If pathogenic bacteria of intestinal
origin are present, coliform
bacteria also are present, in much
greater numbers'.
d Coliforms are always present in the
intestines of humans and other warm-
blooded animals, and are eliminated'
in large numbers in fecal wastes.
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Bacteriological Indicators of Water Pollution
e Coliforms are more persistent in
the aquatic environment than are
pathogenic bacteria of intestinal
origin.
f Coliforms are generally harmless
to humans and can be determined
quantitatively by routine laboratory
procedures.
2 Limitations
a Some of the constituents of the
coliform group have a wide environ-
mental distribution in addition to
their occurrence in the intestines
of warm-blooded animals.
b Some strains of the coliform group
may multiply in certain polluted
waters ("aftergrowth"), of high
nutritive values thereby adding to
the difficulty of evaluating a pollution
situation in the aquatic environment.
Members of the A. aerogenes section
of the coliform are commonly
involved in this kind of problem.
c Because of occasional aftergrowth
problems, the age of the pollution
may be difficult to evaluate under
some circumstances.
d Tests for coliforms are subject to
interferences due to other kinds of
bacteria. False negative results
sometimes occur when species of
Pseudomonas are present. False
positive results sometimes occur
when two or more kinds of non-
conforms produce gas from lactose,
when neither can do so alone
(synergism).
B The Fecal Coliform Component of the
Coliform Group (as determined by elevated
temperature test)
1 Merits
a The majority (over 95% of the coli-
form bacteria from intestines of
warm-blooded animals grow at the
elevated temperature.
b These organisms are of relatively
infrequent occurrence except in
association with fecal pollution.
c Survival of the fecal coliform group
is shorter in environmental waters
than for the coliform group as
whole. It follows, then, that high
densities of fecal coliforms is
indicative of relatively recent
pollution.
d Fecal coliforms generally do not
multiply outside the intestines of
warm-blooded animals. In certain
high-carbohydrate wastes, such as
from the sugar beet refineries,
exceptions have been noted.
2 Limitations
a Feces from warm-blooded animals
include some (though proportionately
low) numbers of coliforms which do
not yield a positive fecal coliform
test when the elevated temperature
test is used as the criterion of
differentiation These organisms
are E colx varieties by present
taxonomic classification.
b There is at present no established
and consistent correlation between
ratios of total coliforms/fecal
coliforms in interpreting sanitary
quality of environmental waters.
In domestic sewage, the fecal
coliform density commonly is
greater than 90% of the total
coliform density. In environmental
waters relatively free from recent
pollution, the fecal coliform density
may range from 10-30% of the total
coliforms. There are, however,
too many variables relating to
water-borne wastes and surface
water runoff to permit sweeping
generalization on the numerical
relationships between fecal- and
total coliforms.
c At this time, evaluations are
underway regarding the survival
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Bacteriological Indicators of Water Pollution
of fecal coliforms in polluted waters
compared with that of enteric
pathogenic bacteria. In recent
pollution studies, species of
Salmonella have been found in the
presence of 220 fecal coliforms per
100 ml (Spino), and 110 fecal
coliforms per 100 ml (Brezenski,
Raritan Bay Project).
V APP LICATIONS OF COLIFORM TESTS
A Current Status in Official Tests
1 The coliform group is designated, in
"Standard Methods for the Examination
of Water and Wastewater" (13th ed.,
1971), through the Completed Test
MPN procedure as the official test
for bacteriological potability of water.
The Confirmed Test MPN procedure
is accepted where it has been demon-
strated, through comparative tests,
to yield results equivalent to the
Completed Test. The membrane filter
method also is accepted for examination
of waters subject to interstate regulation.
2 The 12th edition of Standard Methods
introduced the standard test for fecal
coliform bacteria. It is emphasized
that this is to be used in pollution
studies, and does not apply to the
evaluation of water for potability.
This procedure has been carried to
the 13th Edition.
B Applications
1 Tests for the coliform group as a
whole are used in official tests to
comply with interstate drinking water
standards, state standards for shell-
fish waters, and in most, if not all,
cases where bacterial standards of
water quality have been established
for such use as in recreational or
bathing waters, water supplies, or
industrial supplies. Laboratory
personnel should be aware of possible
implementation of the fecal coliform
group as the official test for recreational
and bathing waters.
2 The fecal coliform test has application
in water quality surveys, as an adjunct
to determination of total coliform
density. The fecal coliform test is
being used increasingly in all water
quality surveys.
3 It is emphasized that no responsible
worker advocates substitution of a
fecal coliform test for total coliforms
in evaluating drinking water quality.
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Bacteriological Indicators of Water Pollution
Part 3. The Fecal Streptococci
I INTRODUCTION
Investigations regarding streptococci
progressed from the streptococci of medical
concern to those which were distributed in
differing environmental conditions which,
again, related to the welfare of man. The
streptococci were originally reported by Laws
and Andrews (1894), and Houston (1899, 1900)
considered those streptococci, which we now
call "fecal streptococci, " as ... "indicative
of dangerous pollution, since they are readily
demonstrable in waters recently polluted and
seemingly altogether absent from waters above
suspicion of contamination.
From their discovery to the present time the
fecal streptococci appear characteristic of
fecal pollution, being consistently present in
both the feces of all warm-blooded animals
and in the environment associated with animal
discharges. As early as 1910 fecal strepto-
cocci were proposed as indicators to the
Metropolitan Water Board of London.
However, little progress resulted in the
United States until improved methods of
detection and enumeration appeared after
World War II.
Renewed interest in the group as indicators
began with the introduction of azide dextrose
broth in 1950, (Mallmann & Seligmann, 1950).
The method which is in the current edition
of Standard Methods appeared soon after.
(Litsky, et al. 1955).
With the advent of improved methods for
detection and enumeration of fecal strep-
tococci, significant body of technical
literature has appeared.
This outline will consider the findings of
various investigators regarding the fecal
streptococci and the significance of discharges
of these organisms into the aquatic environment.
The terms "enterococci, " "fecal
streptococci, " "Group D streptococci,11
"Streptococcus fecalis, " and even
' streptococci have been used in a loose
and interchangeable manner to indicate
the streptococci present m the enteric
tract of warm-blooded animals or of the
fresh fecal material excreted therefrom.
Enterococci are characterized by specific
taxonomic biochemistry. Serological
procedures differentiate the Group D
streptococci from the various groups.
Although they overlap, the three groups,
fecal streptococcus, enterococcus, and
Group D streptococcus, are not synonymous.
Because our emphasis is on indicators of
unsanitary origin, fecal streptococcus is
the more appropriate term and will include
the enterococcus as well as other groups.
A rigid definition of the fecal streptococcus
group is not possible with our present
knowledge. The British Ministry of Health
(1956) defines the organisms as "Gram-
positive" cocci, generally occurring in
pairs or short chains, growing in the
presence of bile salt, usually capable of
development at 45° C, producing acid but
not gas in mannitol and lactose, failing to
attack raffinose, failing to reduce nitrate
to nitrite, producing acid in litmus milk
and precipitating the casein m the form of
a loose, but solid curd, and exhibiting a
greater resistance to heat, to alkaline
conditions and to high concentrations of
salt than most vegetative bacteria. "
However, it is pointed out that "streptococci
departing in one or more particulars from
the type species cannot be disregarded
in water. "
For the proposes of this outline, and in line
with the consensus of most water micro-
biologists in this country, the definition
of the fecal streptococci is-
H FECAL MATERIALS
A Definition
"The group composed of Group D
species consistently present in
significant numbers in fresh fecal
excreta of warm-blooded animals,
which includes all of the enterococcus
group in addition to other groups of
streptococci."
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Bacteriological Indicators of Water Pollution
B Species Isolated
1 Findings
a Human feces
Examination of human fecal specimens
yields a high percentage of the
enterococcus group and usually
demonstration of the S. salivarius
which is generally considered a
member of the human throat flora
and to be surviving in human fecal
materials rather than actively
multiplying in the enteric tract.
Also present would be a small
percentage of variants or biotypes
of the enterococcus group.
b Nonhuman Feces
1) Fecal material which are from
nonhuman or not from fowl will
yield high percentages of the
S. bovis and/or S. equinus
organisms with a concomitantly
reduced percentage of the
enterococcus group.
2) Fowl excreta
Excrement from fowl characteris-
tically yields a large percentage
of enterococcal biotypes as well
as a significant percentage of
enxerococcus group.
2 Significance
Species associations with particular
animal hosts is an established fact and
leads to the important laboratory
technique of partition counting of colonies
from the membrane filter or pour
agar plates in order to establish or
confirm the source of excretal
pollution in certain aquatic investi-
gations .
It is important to realize that a suitable
medium is necessary m order to
allow all of the streptococci which
we consider to be fecal streptococci
to grow in order to give credence to
the derived opinions. Use of liquid
growth media into which direct
inoculations from the sample are
made have not proven to be successful
for partition counting due to the differing
growth rates of the various species of
streptococci altering the original
percentage relationships. Due to the
limited survival capabilities of some
of the fecal streptococci it is necessary
to sample fresh fecal material or water
samples in close proximity to the
pollution source especially when
multiple sources are contributing to a
reach of water. Also the pH range
must be within the range of 4.0-9.0.
Ill FECAL STREPTOCOCCI IN THE
AQUATIC ENVIRONMENT
A General
From the foregoing it is apparent that
the preponderant human fecal streptococci
is composed of the enterococcus group
and, as this is the case, several media
presently available which will detect only
the enterococcal group will be suitable
for use with aquatic samples which are
known to be contaminated or potentially
contaminated with purely domestic
(human) wastes. On the other hand,
when it is known or suspected that other-
than-human wastes have potential egress
to the aquatic environment under investi-
gation, it is necessary to utilize those
media which are capable of quantitating
the whole of the fecal streptococci group.
B Stormwaters and Combined Sewers
1 General
Storm sewers are a series of pipes
and'conduits which receive surface
runoffs from the action of rainstorms
and do not include sewage which are
borne by a system of sanitary sewers.
Combined sewers receive both the runoff
rains as well as the water borne wastes
of the sanitary system. Both of these
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Bacteriological Indicators of Water Pollution
types of runoffs can be discharged to
the aquatic environment and the usual
instance where this occurs, with respect
to the combined sewers, is when the
amount of flow is ui excess of the
amounts capable of beingitreated during
the high flow conditions. Both of these
discharge forms have been found to
usually contain large quantities of
fecal streptococci and in numbers
which generally are larger than that
of the fecal coliform indicator organism.
Stormwaters can be concluded to
represent a typical stream environment
with respect to the presence of chemical
constituents and show a wide range of
electrolytes which at times simulate
that of irrigation waters.
2 Bacteriological Findings
Table 1 represents, in a modified form,
some of the findings of Geldreich and
Kenner (1969) with respect to the
densities of fecal streptococci when
considering Domestic sewage in contrast
to Stormwaters:
Table 1
DISTRIBUTION OF FECAL STREPTOCOCCI
IN DOMESTIC SEWAGES AND STORMWATER
RUNOFFS
Fecal Streptococci
per 100 ml Ratio
Water Source median values FC/FS
Domestic Sewage
Preston, ID
64,
000
5.
3
Fargo, ND
290,
000
4.
5
Moorehead, MN
330,
000
4.
9
Cincinnati, OH
2,470,
000
4.
4
Lawrence, MA
4, 500,
000
4.
0
Monroe, MI
700,
000
27.
9
Denver, CO
2, 900,
000
16.
9
Stormwater
Business District 51,000 0.26
Residential 150, 000 0. 04
Rural 58, 000 0.05
The Ratio FC/FS is that of the
Fecal coliform and Fecal streptococci
and it will be noted that m each case,
when considering the Domestic
Sewage, it is 4. 0 or greater while
it is less than 0. 7 for stormwaters.
The use of this ratio is useful to
identify the source of pollution as
Table 2. ESTIMATED PER CAPITA CONTRIBUTION OF INDICATOR MICROORGANISMS
FROM SOME ANIMA LS*
Average indicator Average contribution
density per gram per capita per 24 hr
of feces
Animals
Avg wt of
Feces/24 hr,
wet wt, g
Fecal
coliform,
million
Fecal
streptococci,
million
Fecal
coliform,
million
Fecal
streptococci,
million
Ratio
FC/FS
Man
150
13.0
3.0
2, 000
450
4.4
Duck
336
33.0
54.0
11, 000
18, 000
0.6
Sheep
1, 130
16.0
38.0
18, 000
43, 000
0. 4
Chicken
182
1.3
3.4
240
620
0.4
Cow
23,600
0.23
1.3
5, 400
31, 000
0.2
Turkey
448
0.29
2.8
130
1, 300
0. 1
Pig
2, 700
3.3
84.0
8,900
230, 000
0.04
^Publication WP-20-3, P. 102
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Bacteriological Indicators of Water Pollution
being human or nonhuman warm-
blooded animal polluted. When the ratio
is greater than 4. 0 it is considered to be
human waste contaminated while a ratio
of less than 0.7 is considered to be
nonhuman. It is evident that the storm-
waters have been primarily polluted by
excreta of rats and other rodents and
possibly domestic and/or farm animals.
Species differences are the main cause
of different fecal coliform-fecal
streptococci ratios. Table 2 compares
fecal streptococcus and fecal coliform
counts for different species. Even
though individuals vary widely, masses
of individuals in a species have charac-
teristic proportion of indicators.
C Surface Waters
In general, the occurrence of fecal
streptococci indicates fecal pollution and
its absence indicates that little or no
warm-blooded fecal contribution. In
studies of remote surface waters the fecal
streptococci are infrequently isolated and
occurrences of small numbers can be
attributed to wild life and/or snow melts
and resultant drainage flows.
Various examples of fecal streptococcal
occurrences are shown in Table 3 m
relation to surface waters of widely varying
quality. (Geldreich and Kenner 1969)
IV FECAL STREPTOCOCCI: ADVANTAGES
AND LIMITATIONS
A General
Serious studies concerning the streptococci
were instituted when it became apparent
that they were the agents responsible or
suspected for a Wide variety of human
diseases. Natural priority then focused
itself to the taxonomy of these organisms
and this study is still causing consternation
as more and more microbiological techniques
have been brought to bear on these questions.
The sanitary microbiologist is concerned
with those streptococci which inhabit the
enteric tract of warm-blooded animals,
their detection, and utilization m develop-
ing a criterium for water quality standards.
Table 3
INDICATOR ORGANISMS IN SURFACE
WATERS
Densities/100 ml
Fecal Fecal
Water Source coliform streptococci
Prairie Watersheds
Cherry Creek, WY 90 83
Saline River, KS 95 180
Cub River, ID 110 160
Clear Creek, CO 170 110
Recreational Waters
Lake Mead 2 444
Lake Moovalaya 9 170
Colorado River 4 256
Whitman River 32 88
Merrimack River 100 96
Public Water Intakes
Missouri River (1959)
Mile 470.5 11,500 39, 500
Mile 434.5 22,000 79, 000
Mile 408.8 14,000 59,000
Kabler (1962) discussed the slow acceptance
of the fecal streptococci as indicators of
pollution resulting from:
1 Multiplicity and difficulty of laboratory
procedures
2 Poor agreement between methods of
quantitative enumeration
3 Lack of systematic studies of ... .
a sources
b survival, and
c interpretations, and
4 Undue attention to the S. faecalis group.
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Bacteriological Indicators of Water Pollution
Increased attention to the fecal streptococci,
especially during the last decade, have
clarified many of the earlier cloudy issues
and have elevated the stature of these
organisms as indicators of pollution.
Court precedents establishing legal status
and recommendations of various technical
advisory boards have placed the fecal
coliform group in a position of primacy
in many water quality applications. The
fecal streptococci have evolved from a
position of a theoretically useful indicator
to one which was ancillary to the coliforms
to one which was useful when discrepancies
or questions evolved as to the validity of
the coliform data to one where an equality
status was achieved in certain applications.
In the future it is anticipated that, for
certain applications, the fecal streptococci
will achieve a position of primacy for
useful data, and, as indicated by Litsky
(1955) "be taken out of the realm of step-
children and given their legitimate place
m the field of santiary bacteriology as
indicators of sewage pollution. "
B Advantages and Limitations
1 Survival
In general, the fecal streptococci have
been observed to have a more limited
survival time in the aquatic environment
when compared to the coliform group.
They are rivaled in this respect only
by the fecal coliforms. Except for cases
of persistence in waters of high electro-
lytic content, as may be common to
irrigation waters, the fecal streptococci
have not been observed to multiply in
polluted waters as may sometimes be
observed for some of the coliforms.
Fecal streptococci usually require a
greater abundance of nutrients for sur-
vival as compared to the coliforms and
the coliforms are more dependent upon
the oxygen tension in the waterbody.
In a number of situations it was concluded
that the fecal streptococci reached an
extinction point more rapidly in warmer
waters while the reverse was true in the
colder situations as the coliforms now
were totally eliminated sooner.
2 Resistance to Disinfection
In artificial pools the source of
contamination by the bathers is
usually limited to throat and skin
flora and thus increasing attention
has been paid to indicators other
than those traditionally from the
enteric tract. Thus, one of the
organisms considered to be a fecal
streptococci, namely, S. salivarius,
can be a more reliable indicator
when detected along with the other
fecal streptococci especially since
studies have confirmed the greater
resistance of the fecal streptococci
to chlorination. This greater
resistance to chlorination, when
compared to the fecal coliforms, is
important since the dieoff curve
differences are insignificant when
the curves of the fecal coliforms
are compared to various Gram
negative pathogenic bacteria which
reduces their effectiveness as
indicators.
3 Ubiquitous Strains
Among the fecal streptococcus are
two organisms, one a biotype and
the other a variety of the S. faecalis,
which, being ubiquitous (omnipresent)
have limited sanitary significance.
The biotype, or atypical, S. faecalis
is characterized by its ability to
hydrolyze starch while the varietal
form, liquefaciens, is nonbeta
haemolytic and capable of liquefying
gelatin. Quantitation of these organisms
in anomalous conditions is due to their
capability 'of survival in soil or high
electrolytic waters and in waters with
a temperature of less than 12 Degrees C.
Samples have been encountered which
have been devoid of fecal coliforms
and yet contain a substantial number of
"fecal streptococci" of which these
ubiquitous strains constitute the majority
or all of the isolations when analyzed
biochemically.
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Bacteriological Indicators of Water Pollution
V STANDARDS AND CRITERIA
Acceptance and utilization of Total Coliform
criteria, which must now be considered a
pioneering effort, has largely been supplanted
in concept and in fact by the fecal coliforms
in establishing standards for recreational
waters.
The first significant approach to the utiliza-
tion of the fecal streptococci as a criterium
for recreational water standards occurred in
1966 when a technical committee recommended
the utilization of the fecal streptococci with the
total coliforms as criteria for standards
pertaining to the Calumet River and lower
Lake Michigan waters. Several sets of
criteria were established to fit the intended
uses for this area. The use of the fecal
streptococci as a criterium is indicated to
be tentative pending the accumulation of
existing densities and could be modified in
future standards.
With the existing state-of-the-art knowledge
of the presence of the fecal streptococci in
waters containing low numbers of fecal
coliforms it is difficult to establish a specific
fecal streptococcus density limit of below
100 organisms/100 ml when used alone or
in conjunction with the total coliforms.
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Bacteriological Indicators of Water Pollution
Part 4. Other Batterlal Indicators of Pollution
I TOTAL BACTERIA L COUNTS
A Historical
1 The early studies of Robert Koch led
him to develop tentative standards of
water quality based on a limitation of
not more than 100 bacterial colonies
per ml on a gelatin plating medium
incubated 3 days at 20OC.
2 Later developments led to inoculation
of samples on duplicate plating media,
with one set incubated at 370 c and the
other at 20° C.
a Results were used to develop a ratio
between the 37° C counts and the
20C>C counts.
b Waters having a predominant
count at 37° C were regarded as
being of probable sanitary signifi-
cance, while those giving
predominant counts at 20° C were
considered to be of probable soil
origin, or natural inhabitants of
the water being examined.
B Groups Tested
There is no such thing as "total" bacterial
count in terms of a laboratory determination.
1 Direct microscopic counts do not
differentiate between living and dead
cells.
2 Plate counting methods enumerate only
the bacteria which are capable of using
the culture medium provided, under the
temperature and other growth conditions
used as a standard procedure. No one
culture medium and set of growth
conditions can provide, simultaneously,
an acceptable environment for all the
heterogeneous, often conflicting,
requirements of the total range of
bacteria which may be recovered from
waters.
C Utilization of Total Counts
1 Total bacterial counts, using plating
methods, are useful for:
a Detection of changes in the bacterial
composition of a water source
b Process control procedures in
treatment plant operations
c Determination of sanitary conditions
in plant equipment or distributional
systems
2 Serious limitations in total bacterial
counts exist because.
a No information is given regarding
possible or probable fecal origin
of bacterial changes. Large numbers
of bacteria can sometimes be
cultivated from waters known to be
free of fecal pollution.
b No information of any kind is given
about the species of bacteria
cultivated.
c There is no differentiation between
harmless or potentially dangerous
forms.
3 Status of total counts
a There is no total bacterial count
standard for any of the following
Interstate Quarantine Drinking
Water Standards
PHS regulations for water
potability (as shown in
"Standard Methods" Public
Health Service Drinking
Water Standards of 1962.)
b The most widely used current
application of total bacterial counts
in water bacteriology today is in
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Bacteriological Indicators of Water Pollution
water treatment plants, where some
workers use standard plate counts
for process control and for deter-
mination of the bacterial quality of
distribution systems and equipment.
c Total bacterial counts are not used
in PHS water quality studies, though
extensively used until the 1940's.
B Spore-Forming Bacteria (Clostridium
perfringens, or C. welchii)
1 Distribution
This is one of the most widely distributed
species of bacteria. It is regularly
present in the intestinal tract of warm-
blooded animals.
2 Nature of organism
C. perfringens is a Gram-positive,
spore-forming rod. The spores cause
a distinct swelling of the cell when
formed. The organism is extremely
active in fermentation of carbohydrates,
and produces the well-known "stormy
fermentation" of milk.
3 Status
The organism, when present, indicates
that pollution has occurred at some
time. However, because of the ex-
tremely extended viability of the spores,
it is impossible to obtain even an
approximation of the recency of pollution
based only on the presence of
C. perfringens.
The presence of the organism does not
necessarily indicate an unsafe water.
C Tests for Pathogenic Bacteria of Intestinal
Origin
1 Groups considered include Salmonella
sp, Shigella sp, Vibrio comma,
Mycobacterium sp, Pasteurella sp,
Leptospira sp, and others.
2 Merits of direct tests
Demonstration of any pathogenic
species would demonstrate an
unsatisfactory water quality, hazardous
to persons consuming or coming into
contact with that water.
3 Limitations
a There is no available routine pro-
cedure for detection of the full
range of pathogenic bacteria cited
above.
b Quantitative methods are not avail-
able for routine application to any
of the above.
c The intermittent release of these
pathogens makes it impossible to
regard water as safe, even in the
absence of pathogens.
d After detection, the public already
would have been exposed to the
organism, thus, there is no built-in
margin of safety, as exists with
tests for the coliform group.
4 Applications
a In tracing the source of pathogenic
bacteria in epidemiological investi-
gations
b In special research projects
c In water quality studies concerned
with enforcement actions against
pollution, increasing attention is
being given to the demonstration of
enteric pathogenic bacteria in the
presence of the bacterial indicators
of pollution.
D Miscellaneous Indicators
It is beyond this discussion to explore the
total range of microbiological indicators
of pollution that have been proposed and
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Bacteriological Indicators of Water Pollution
investigated to some extent. Mention can
be made, however, of consideration of
tests for the following.
1 Bacteriophages specific for any of a
number of kinds of bacteria
2 Serological procedures for detection
of coliforrns and other indicators, a
certain amount of recent attention has
been given to applications of fluorescent
antibodies in such tests
3 Tests for Pseudomonas aeruginosa
4 Tests for viruses, which may persist
in waters even longer than members
of the coliform group.
REFERENCES
1 Standard Methods for the Examination of
Water and Wastewater, 13th ed.,
APHA, AWWA, WPCF. Published by
American Public Health Association,
1790 Broadway, New York, N. Y. 1971.
2 Prescott, S. C., Winslow, C.E.A., and
McCrady, M. Water Bacteriology.
John Wiley & Sons, Inc. 1946.
3 Parr, L.W. Coliform Intermediates in
Human Feces. Jour. Bact. 36 1.
1938.
4 Clark, H. F. and Kabler, P.W. The
Physiology of the Coliform Group.
Proceedings of the Rudolfs Research
Conference on Principles and Appli-
cations in Aquatic Microbiology. 1963.
5 Geldreich, E. E., Bordner, R.H., Huff,
C.B., Clark, H.F. , and Kabler, P.W.
Type Distribution of Coliform Bacteria
in the Feces of Warm-Blooded Animals.
JWPCF. 34 295-301. 1962.
6 Geldreich et al. The Fecal Coli-Aerogenes
Flora of Soils from Various Geographic
Areas. Journal of Applied Bacteriology
25.87-93. 1962.
7 Geldreich, E.E., Kenner, B.A., and
Kabler, P.W. Occurrence of
Coliforrns, Fecal Coliforrns, and
Streptococci on Vegetation and Insects.
Applied Microbiology. 12 63-69. 1964.
8 Kabler, P.W., Clark, H.F., and
Geldreich, E.E. Sanitary Significance
of Coliform and Fecal Coliform
Organisms in Surface Water. Public
Health Reports. 79.58-60. 1964.
9 Clark, H.F. and Kabler, P.W.
Re-evaluation of the Significance of the
Coliform Bacteria. Journal AWWA.
56-931-936. 1964.
10 Kenner, B. S., Clark, H.F., and
Kabler, P.W. Fecal Streptococci.
II. Quantification in Feces. Am. J.
Public Health. 50 1553-59. 1960.
11 Litsky, W., Mailman, W.L., and Fifield,
C. W. Comparison of MPN of
Escherichia coli and Enterococci in
River Water. Am. Jour. Public Health.
45 1949. 1955.
12 Medrek, T.F. and Litsky, W.
Comparative Incidence of Coliform
Bacteria and Enterococci in
Undisturbed Soil. Applied Micro-
biology. 860-63. 1960.
13 Mailman, W.L., and Litsky, W.
Survival of Selected Enteric Organisms
in Various Types of Soil. Am. J.
Public Health. 41'38-44. 1950.
14 Mailman, W.L., and Seligman, E.B., Jr.
A Comparative Study of Media for
Detection of Streptococci in Water and
Sewage. Am J Public Health.
40 286-89. 1950.
15 Ministry of Health (London). The
Bacterial Examination of Water Supplies.
Reports on Public Health and Medical
Subjects. 71 34.
-------
Bacteriological Indicators of Water Pollution
16 Morris, W. and Weaver, R.H.
Streptococci as Indices of Pollution
in Well Water. Applied Microbiology.
2:282-285. 1954.
17 Mundt, J.O., Coggin, J.H., Jr., and
Johnson, L.F. Growth of
Streptococcus fecalis var. liquefaciens
on Plants. Applied Microbiology.
10:552-555. 1962.
18 Geldreich, E. E. Sanitary Significance
of Fecal Coliforms in the Environment.
U. S. Department of the Interior.
FWPCA Publ. WP-20-3. 1966.
19 Geldreich, E. E. and Kenner, B. A.
Concepts of Fecal Streptococci in
Stream Pollution. J.WPCF. 41.-R336.
1969.
20 Kabler, P. W. Purification and Sanitary
Control of Water {Potable and Waste)
Ann. Rev. of Microbiol. 16:127. 1962.
21 Litsky, W., Mailman, W. L., and Fifield,
C. W. Comparison of the Most Probable
Numbers of Escherichia coli and
Enterococci in River Waters. A. J.P.H.
45:1049. 1955.
22 Geldreich, E. E. Applying Bacteriological
Parameters to Recreational Water Quality.
J.AWWA. 62:113. 1970.
23 Geldreich, E. E., 'Best, L. C., Kenner, B. A.
and Van Donsel, D. J. The Bacteriolog-
ical Aspects of Stormwater Pollution.
J.WPCF. 40:1860. 1968.
24 FWPCA Report of Water Quality Criteria
Calumet Area - Lower Lake Michigan,
Chicago, IL. Jan. 1966.
This outline was prepared by H. L. Jeter,
Director, National Training Center and
revised by R. Russomanno, Microbiologist,
National Training Center, WPO, EPA,
Cincinnati, OH 45268.
-------
FUNGI AND THE "SEWAGE FUNGUS" COMMUNITY
I INTRODUCTION
A Description
Fungi are heterotrophicachylorophyllous
plant-like organisms which possess true
nuclei with nuclear membranes and nu-
cleoli. Dependent upon the species and
in some instances the environmental
conditions, the body of the fungus, the
thallus, varies from a microscopic
single cell to an extensive Plasmodium
or mycelium. Numerous forms produce
macroscopic fruiting bodies.
B Life Cycle
The life cycles of fungi vary from simple
to complex and may include sexual and
asexual stages with varying spore types
as the reproductive units.
C Classification
Traditionally, true fungi are classified
within the Division Eumycotina of the
Phylum Mycota of the plant kingdom.
Some authorities consider the fungi an
essentially monophyletic group distinct
from the classical plant and animal
kingdoms.
II ACTIVITY
In general, fungi possess broad enzymatic
capacities. Various species are able to
actively degrade such compounds as
complex polysaccharides (e.g., cellulose,
chitin, and glycogen), proteins (casein,
albumin, keratin), hydrocarbons (kerosene)
and pesticides. Most species possess an
oxidative or microaerophilic metabolism,
but anaerobic catabolism is not uncommon.
A few species show anaerobic metabolism
and growth.
Ill ECOLOGY
A Distribution
Fungi are ubiquitous in nature and mem-
bers of all classes may occur in large
numbers in aquatic habitats. Sparrow
(1968) has briefly reviewed the ecology
of fungi in freshwaters with particular
emphasis on the zoosporic phycomycetes.
The occurrence and ecology of fungi in
marine and estuarine waters has been
examined recently by a number of in-
vestigators (Johnson and Sparrow, 1961,
Johnson, 1968, Myers, 1968, van Uden
and Fell, 1968).
B Relation to Pollution
Wm. Bridge Cooke, in a series of in-
vestigations (Cooke, 1965), has estab-
lished that fungi other than phycomycetes
occur in high numbers in sewage and
polluted waters. His reports on organic
pollution of streams (Cooke, 1961, 1967)
show that the variety of the Deuteromy-
cete flora is decreased at the immediate
sites of pollution, but dramatically in-
creased downstream from these regions.
Yeasts, in particular, have been found
in large numbers in organically enriched
waters (Cooke, et al., 1960, Cooke and
Matsuura, 1963, Cooke, 1965b; Ahearn,
et al., 1968). Certain yeasts are of
special interest due to their potential
use as "indicator" organisms and their
ability to degrade or utilize proteins,
various hydrocarbons, straight and
branch chained alkyl-benzene sulfonates,
fats, metaphosphates, and wood sugars.
BI. FU. 6a. 5. 71
-------
Fungi
C "Sewage Fungus" Community (Plate I)
A few microorganisms have long been
termed "sewage fungi. " The most
common microorganisms included in
this group are the iron bacterium
Sphaerotilus natans and the phycomy-
cete Leptomitus lacteus.
1 Sphaerotilus natans is not a fungus,
rather it is a sheath bacterium of
the order chlamydobacteriales.
This polymorphic bacterium occurs
commonly in organically enriched
streams where it may produce
extensive slimes.
a Morphology
Characteristically, S. natans
forms chains of rod shaped
cells (1. 1-2. On x 2.5- 17n)
within a clear sheath or tri-
chome composed ofaprotein-
polysaccharidae-lipid complex.
The rod cells are frequently
motile upon release from the
sheath; the flagella are lopho-
trichous. Occasionally two
rows of cells may be present
in a single sheath. Single tn-
chomes may be several mm
in length and bent at various
angles. Empty sheaths, ap-
pearing like thin cellophane
straws, may be present
b Attached growths
The trichomes are cemented
at one end to solid substrata
such as stone or metal, and
their cross attachment and
bending gives a superficial
similarity to true fungal hyphae.
The ability to attach firmly to
solid substrates gives S. natans
a selective advantage in the
population of flowing streams.
For more thorough reviews of
S.natans see Prigsheim(1949)
and Stokes (1954).
2 Leptomitus lacteus also produces
extensive slimes and fouling floes
in fresh waters. This species forms
thalli typified by regular constrictions.
a Morphology
Cellulin plugs may be present
near the constrictions and there
may be numerous granules in
the cytoplasm. The basal cell
of the thallus may possess
rhizoids.
b Reproduction
The segments delimited by the
partial constrictions are con-
verted basipetally to sporangia.
The zoospores are diplanetic
(l. e., dimorphic) and each
possesses one whiplash and one
tinsel flagellum. No sexual
stage has been demonstrated
for this species.
c Distribution
For further information on the
distribution and systematics
of L. lacteus see Sparrow (1960),
Yerkes (1966) and Emersonand
Weston (1967). Both S. natans
and L. lacteus appear to thrive
in organically enriched cold
waters (5°-22°C) and both seem
incapable of extensive growth at
temperatures of about 30°C.
d Gross morphology
Their metabolism is oxidative
and growth of both species may
appear as reddish brown floes
or stringy slimes of 30 cm or
more in length.
e Nutritive requirements
Sphaerotilus natans is able to
utilize a wide variety of organic
compounds, whereas L. lacteus
does not assimilate simple
-------
Fungi
PLATE I
'SEWAGE FUNGUS" COMMUNITY OR "SLIME GROWTHS"
(Attached "filamentous" and slime growths)
Zoogloea
Sphaerotilus natana
Beggiatoa alba
Sm
BACTERIA
Fusarium aqueJuctum
// Epistylia 8
Leptomitus lacteus
Geotrichum candidum
Carchesium
-t> *
FUNGI
/OO
10
Opercularia
PROTOZOA
-------
Fungi
PLATE II
REPRESENTATIVE FUNGI
Figure «*¦
Fusarium aquaeductuum
(Radlmacher and
Rabenhorst) Saccardo
Microconidia (A) produced
from phialides at in Cephalo-
sporium, remaining in slime
balls. Macroconidia (B), with
one to several cross walls,
produced from collared phial-
ides. Drawn from culture.
Figure 2.
Leptcmitus lacteus (Roth)
Agardh
Cells of the hyphae show-
ing constrictions with cellulin
plugs. In one cell large zoo-
spore* have been delimited.
Redrawn from Coker, 1923.
Figure 4"
Zoophagus insidians
Sommerstorff
Mycelium with hyphal pegs
\ (A) on which rotifers will
\ become impaled; gemmae (B)
\ produced as conidia on short
\ J hyphal branches; and rotifer
impaled on hyphal peg (C)
\ I . . from which hyphae have
/ grown into the rotifer whose
shell will be discarded after
the contents are consumed.
f\ Drawn from culture.
Figure 3
Geotrichum candidum
Link ex Persoon
Mycelium with short cells
and arthrospores. Young hy-
pha (A); and mature arthro-
spores (B). Drawn from col-
Figure 5"
Achlya americana Humphrey
Ooogonium with three oo-
spores (A); young zoospor-
angium with delimited zoo-
spores (B); and zoosporangia
(C) with released zoospores
that remain encysted in clus-
ters at the mouth of the dis-
charge tube. Drawn from cul-
ture.
Ihr inoci/xtidin m in a riu u in
Figure f Haplosporidium costale. A—mature &poiv
B—early Plasmodium.
Figures 1 through 5 from Cooke; Figures 6 and 7 from Galtsoff.
-------
Fungi
sugars and grows most luxuriantly in
the presence of organic nitrogenous
wastes.
3 Ecological roles
Although the "sewage fungi" on
occasion attain visually noticeable
concentrations, the less obvious
populations of deuteromycetes may
be more important in the ecology of
the aquatic habitat. Investigations of
the past decade indicate that numerous
fungi are of primary importance in the
mineralization of organic wastes, the
overall significance and exact roles of
fungi in this process are yet to be
established.
IV CLASSIFICATION
In recent classification schemes, classes
of fungi are distinguished primarily on the
basis of the morphology of the sexual and
zoosporic stages. In practical schematics,
however, numerous fungi do not demonstrate
these stages. Classification must therefore
be based on the sum total of the morphological
and/or physiological characteristics. The
extensive review by Cooke (1963) on methods
of isolation and classification of fungi from
sewage and polluted waters precludes the
need herein of extensive keys and species
illustrations. A brief synopsis key of the
fungi adapted in part from Alexopholous
(1962) is presented on the following pages.
D Predacious Fungi
1 Zoophagus msidians
(Plate II, Figure 4) has been observed
to impair functioning of laboratory
activated sludge units (see Cooke and
Ludzack).
2 Arthrobotrys is usually found along
with Zoophagus in laboratory activated
sludge units. This fungus is predacious
upon nematodes. Loops rather than
"pegs" are used in snaring nematodes.
PLATE II (Figure 4)
This outline was prepared by Dr. Donald G.
Ahearn, Professor of Biology, Georgia State
College, Atlanta, Georgia 30303.
-------
Fungi
KEY TO THE MAJOR TAXA OF FUNGI
1 Definite cell walls lacking, somatic phase a free living Plasmodium
Sub-phylum Myxomycotina (true slime molds) Class Myxomycetes
1' Cell walls usually well defined, somatic phase nof a free-living Plasmodium
(true fungi) Sub-phylum Eumycotina 2
2 Hyphal filaments usually coenoctytic, rarely septate, sex cells when present forming
oospores or zygospores, aquatic species propagating asexually by zoospores, terrestrial
species by zoospores, sporangiospores conidia or conidia-like sporangia "Phycomycetes" 3
The phycomycetes are generally considered to include the most primitive of the true
fungi As a whole they encompass a wide diversity of forms with some showing relation-
ships to the flagellates, while others closely resemble colorless algae, and still others
are true molds The vegetative body (thallus) may be non-specialized and entirely con-
verted into a reproductive organ (holocarpic), or it may bear tapering rhizoids, or be
mycelial and very extensive The outstanding characteristics of the thallus is a tendency
to be nonseptate and, in most groups, rnultinucliate, cross walls are laid down in vigorously
growing materia) only to delimit the reporductive organs The spore unit of nonsexual re-
production is borne in a sporangium, and, in aquatic and semiaquatic orders, is provided
with a single posterior or anterior flagellum or two laterally attached ones Sexual activity
in the phycomycetes characteristically results in the formation of resting spores
2' (D Hyphal filaments when present septate, without zoospores, with or without sporangia,
usually with comda, sexual reproduction absent or culminating in the formation of asci
or basidia 8
3 (2) Flagellated cells characteristically produced 4
3' Flagellated cells lacking or rarely produced 7
4 (3) Motile cells uniflagellate ' 5
4' Motile cells biflagellate 6
5 (4) Zoospores posteriorly uniflagellate, formed inside the sporangium class Chytndiomycetes
The Chytndiomycetes produce asexual zoospores with a single posterior whiplash
flagellum The thallus is highly variable, the most primitive forms are unicellular and
holocarpic and in their early stages of development are plasmodial flack cell walls), more
advanced forms develop rhizoids and with further evolutionary progress develop mycelium
The principle chemical component of the cell wall is chitin, but cellulose is also present
Chytrids are typically aquatic organisms but may be found in other habitats Some species
are chitinolytic and/or keratinolytic Chytrids may be isolated from nature by baiting (e g
hemp seeds or pine pollen) Chytrids occur both in marine and fresh water habitats and are
of some economic importance due to their parasitism of algae and animals The genus
DermocYstidium may be provisionally grouped with the chytrids Species of this genus
cause serious epidemics of oysters and marine and fresh water fish
51 Zoospores anteriorly uniflagellate, formed inside or outside the sporangium , , class
Hyphochytndiomycetes
These fungi are aquatic (fresh water or marine) chytrid-hke fungi whose motile cells
possess a single anterior flagellum of the tinsei type (feather-like) They are parasitic on
algae and fungi or may be saprobic Cell walls contain chitin with some species also demon-
strating cellulose content Little information is available on the biology of this class and
at present it is limited to less than 20 species
6 (41) Flagella nearly equal, one whiplash the other tinsel class Oomycetes
A number of representatives of the Oomycetes have been shown to have cellulosic cell
walls The mycelium is coenocytic, branched and well developed in most cases The sexual
process results in the formation of a resting spore of the oogamous type, i e , a type of
fertilization in which two heterogametangia come in contact and fuse their contents through
a pore or tube The thalh in this class range from unicellular to profusely branched
filamentous types Most forms are eucarpic, zoospores are produced throughout the class
except in the more highly advanced species Certain species are of economic importance due
to their destruction of food crops {potatoes and grapes) while others cause serious diseases of
fish (e g Saprolegina parasitica) Members of the family Saprolegmaceae are the common
-------
Fungi
water molds and are among the most ubiquitous fungi in nature The order Lagemdiales
includes only a feu- species which are parasitic on algae, small animals, and other aquatic
life The somatic structures of this taxon are holocarpic and endobiotic The sewage fungi
are classified in the order Leptorrutales Fungi of this order are characterized by the
formation of refractile constrictions 'cellulin plugs ' occur throughout the thalli or, at least,
at the bases of hyphae or to cut off reproductive structures Leptomitus lacteus may
produce rather extensive fouling floes or slimes in organically enriched waters
6' Flagella of unequal size both whiplash class Plasmodiophoromycetes
Members of this class are obligate endoparasites of vascular plants, algae and fungi
The thallus consists of a plasmodium which develops within the host cells Nuclear division
at some stages of the life cycle is of a type found in no other fungi but known to occur in
protozoa Zoosporangia which arise directly from the plasmodium bear zoospores with two
unequal anterior falgella The cell walls of the9e fungi apparently lack cellulose
7 {3') Mainly *saprobic sex cell when present a zygospore class 7 \ gomycetes
This class has well developed mycelium with septa developed in portions of the
older hyphae, actively growing hyphae are normally non-septate The asexual spores are
non-motile sporangiospores (aplanospores) Such spores lack flagella and are usually
aerialy disseminated Sexual reproduction is initiated by the fusion of two gametangia
with resultant formation of a thick-walled, resting spore, the zygospore In the more
advanced species the sporangia or the sporangiospores are conidia-like Many of the
Zygomycetes are of economic importance due to their ability to synthesize commercially
valuable organic acids and alcohols, to transform steroids such as cortisone and to
parasitize and destroy food crops A few species are capable of causing disease in man
and animals (zygomycosis)
7' Obligate commensals of arthropods, zygospores usually lacking class Trichomycetes
The Trichomycetes are an ill-studied group of fungi which appear to be obligate
commensals of arthropods The trichomycetes are associated with a wide variety of insecta
diplopods, and crustacea of terrestrial and aquatic (fresh and marine) habitats None of
the members of this class have been cultured in vitro for continued periods of times with any
success Asexual reproduction is by means of sporangiospores Zygospores have been
observed in species of several orders
8 (2') Sexual spores borne in asci class Ascomycetes
In the Ascomycetes the products of meiosis, the ascospores, are borne in sac
like structures termed asci The ascus usually contains eight ascospores, but the number
produced may vary with the species or strain Most species produce extensive septate
mycelium This large class is divided into two subclasses on the presence or absence
of an ascocarp The Herniascomycetidae lack an ascocarp and do not produce ascogenous
hyphae, this subclass includes the true yeasts The Euascomycetidae usually are divided
into three series (Plectomycetes, Pyrenomycetes, and Discomycetes) on the basis of
ascocarp structure
8' Sexual spores borne on basidia class Basidiomycetes
The Basidiomycetes generally are considered the most highly evolved of the fungi
Karyogamy and meiosis occur in the basidium which bears sexual exogenous spores,
basidiospores The mushrooms toadstools, rusts, and smuts are included in this class
8" Sexual stage lacking ' Form class (Fungi Imperfecti) Deuteromycetes
The Deuteromycetes is a form class for those fungi (with morphological affinities
to the Ascomycetes or Basidiomycetes) which have not demonstrated a sexual stage
The generally employed classification scheme for these fungi is based on the morphology
and color of the asexual reproductive stages This scheme is briefly outlined below
Newer concepts of the classification based on comdium development after the classical
work of S J Hughes (1953) may eventually replace the gross morphology system {see
Barron 1968)
-------
Fungi
KEY TO THE FORM-ORDERS OF THE FUNGI IMPERFECli
I Reproduction by means of conidia, oidia. or by budding 2
II No reproductive structures present Mycelia Sterilia
2 (1) Reproduction by means of conidia borne in pycnidia Sphaeropsidalee
2' Conidia, when formed, not in cycnidia 3
3 (2') Conidia borne m acervuli Melancomales
3' Conidia borne otherwise, or reproduction by oidia or by budding Moniliales
KEY TO THE FORM-FAMILIES OF THE MONILIALES
1 Reproduction mainly by unicellular budding, yeast-like, mycelial phase, if present,
secondary, arthroapores occasionally produced, manifest melanin pigmentation lacking 2
1' Thallus mainly filamentous, dark melanin pigments sometimes produced 3
2 (1) Ballistospores produced Sporobolomycetaceae
2' No ballistospores Cryptococcaceae
3 Conidiophores, if present, not united into sporodochia or synnemata 4
V Sporodochia present Tuberculariaceae
3" Synnemata present Stilbellaceae
4 (3) Conidia and conidiophores or oidia hyaline or brightly colored Moniliaceae
4' Conidia and/or conidiophores, containing dark melanin pigment Dematiaceae
-------
Fungi
SELECTED REFERENCES
Ahearn, D. G , Roth, F.J. Jr., Meyers, S. P.
Ecology and Characterization of Yeasts
from Aquatic Regions of South Florida.
Marine Biology 1 291-308. 1968
Alexopoulos, J.C. Introductory Mycology
2nd ed. John Wiley and Sons, New York,
613 pp 1962
Barron, G L. The Genera of Hyphomycetes
from Soil. Williams and Wilkins Co.,
Baltimore 364 pp. 1968
Cooke, W.B. Population Effects on the
Fungus Population of a Stream.
Ecology 42 1-18. 1961
. A Laboratory Guide to Fungi in
Polluted Waters, Sewage, and Sewage
Treatment Systems U S. Dept. of
Health, Education and Welfare, Cincinnati,
132 pp. 1963
. Fungi in Sludge Digesters.
Purdue Univ Proc. 20th Industrial
Waste Conference, pp 6-17 1965a
. The Enumeration of Yeast
Populations in a Sewage Treatment Plant.
Mycologia 57'696-703. 1965b
Fungal Populations in Relation
to Pollution of the Bear River, Idaho-Utah.
Utah Acad. Proc. 44(1) 298-315. 1967
and Matsuura, George S. A Study
of Yeast Populations in a Waste Stabilization
Pond System. Protoplasma 57 163-187.
1963
, Phaff, H.J., Miller, M.W.,
Shifrine, M., and Knapp, E. Yeasts
in Polluted Water and Sewage.
Mycologia 52 210-230. 1960
Emerson, Ralph and Weston, W.H.
Aqualinderella fermentans Gen. et Sp.
Nov., A Phycomycete Adapted to
Stagnant waters I Morphology and
Occurrence in Nature. Amer. J
Botany 54 702-719. 1967
Hughes, S J Conidiophores, Conidia and
Classification. Can. J. Bot.31 577-
659. 1953
Johnson, T. W., Jr. Saprobic Marine Fungi
pp. 95-104. In Ainsworth, G.C. and
Sussman, A.S. The Fungi, III.
Academic Press, New York. 1968
and Sparrow, F.K., Jr. Fungi
in Oceans and Estuaries. Weinheim,
Germany. 668 pp. 1961
Meyers, S.P. Observations on the Physio-
logical Ecology of Marine Fungi. Bull.
Misaki Mr. Biol. Inst. 12 207-225. 1968
Prigsheim, E.G. Iron Bacteria. Biol. Revs.
Cambridge Phil. Soc. 24.200-245. 1949
Sparrow, F. K. , Jr. Aquatic Phycomycetes.
2nd ed. Univ. Mich, Press, AnnArbor.
1187 pp. 1960.
. Ecology of Freshwater Fungi,
pp. 41-93. In Ainsworth, G.C. and
Sussman, A.S The Fungi, III. Acad.
Press, New York. 1968
Stokes, J. L. Studies on the Filamentous
Sheathed Iron Bacterium Sphaerotilus
natans. J. Bacteriol. 67:278-291. 1954
van Uden, N. and Fell, J.W. Marine Yeasts,
pp. 167-201. In Droop, M.R. and Wood,
E.J.F. Advances in Microbiology of
the Sea, I. Academic Press, New York.
1968
Yerkes, W.D. Observations on an Occurrence
of Leptomitus lacteus in Wisconsin.
Mycologia 58-976-978. 1966
Cooke, William B. and Ludzack, F.J.
Predacious Fungus Behavior in
Activated Sludge Systems. Jour. Water
Poll. Cont. Fed. 30(12) 1490-1495. 1958.
-------
FRESHWATER POLLUTION ECOLOGY
Q.
WHAT IS ECOLOGY?
A.
The science of the interrelation between living
organisms and their environment.
Q.
WHAT IS NOT ECOLOGY?
A.
Not much1
T. T. Macan
SECTION E RESPONSE OF AQUATIC COMMUNITIES
TO CHANGES IN WATER QUA LITY
SECTION F WATER QUA LITY AND AQUATIC LIFE
SECTION G SOME CURRENT POLLUTION PROBLEMS
-------
SECTION E
WATER QUALITY AFFECTS AQUATIC COMMUNITIES
In this section a series of outlines identify typical changes in aquatic com-
munities as a response to a variety of insults. Obvious changes may take
place in population numbers of a single species as well as in community
balance. Biology may be used for the characterization of water quality and
interpretation of population trends within the biota. The final outline is a
review of past and current efforts to relate water quality of biological
communities.
Contents of Section E
Outline No.
Biological Aspects of Natural Self Purification
26
Ecology of Waste Stabilization Processes
26
Effects of Pollution on Fish
27
The Interpretation of Biological Data with Reference
to Water Quality
-------
BIOLOGICAL ASPECTS OF NATURAL SELF PURIFICATION
I INTRODUCTION
A The results of natural self purification
processes are readily observed. Did they
not exist, sewage (and other organic
wastes) would forever remain, and the
world as we know it would long ago have
become uninhabitable. Physical, chemical,
and biological factors are involved. The
microscopic and macroscopic animals
and plants in a body of water receiving
organic wastes are not only exposed to all
of the various (ecological) conditions in
that water, but they themselves create and
profoundly modify certain of those conditions.
B Since toxic chemicals kill some of or all
of the aquatic organisms, their presence
disrupts the natural self purification
processes, and hence, will not be considered
here. The following discussion is based
solely on the effects of organic pollution
such as sewage or other readily oxidizable
organic wastes.
C This description is based on the concept of
a "stream" since under the circumstances
of stream or river flow, the events and
conditions occur in a linear succession.
The same fundamental processes occur in
lakes, estuaries, and oceans, except that
the sequence of events may become
telescoped or confused due to the reduction
or variability of water movements.
D The particular biota (plants and animals,
or flora and fauna) employed as illustrations
below are typical of central United States.
Similar or equivalent forms occur in
similar circumstances in other parts of
the world.
E This presentation is based on an unpublished
chart produced by Dr. C.M. Tarzwell and
his co-workers in 1951. Examples from
this chart are employed m the presentation.
II THE STARTING POINT
A A normal unpolluted stream is assumed
as a starting point. (Figure 1)
B The cycle of life is in reasonably stable
balance.
C A great variety of life is present, but no
one species or type predominates.
D The organisms present are adjusted to the
normal ranges of physical and chemical
factors characteristic of the region, such
as the following
1 The latitude, turbidity, typical cloud
cover, etc. affect the amount of light
penetration and hence photosynthesis.
2 The slope, cross sectional area, and
nature of the bottom affect the rate of
flow, and hence the type of organisms
present deposition of sludge, etc.
3 The temperature affects both certain
physical characteristics of the water,
and the rate of biological activity
(metabolism).
4 Dissolved substances naturally present
in the water greatly affect living
organisms (hard water vs. soft water
fauna and flora).
E Clean water zones can usually be
characterized as follows-
1 General features-
a Dissolved oxygen high
b BOD low
c Turbidity low
d Organic content low
BI. ECO.nap. 5c. ?2. 70
-------
Biological Aspects of Natural Self Purification
THE BIOTA
J
a 30
CLEAN WATER
24 12
CLEAN WATER
3 4
DAYS
12 24 36 48 60 72 84 96 108
MILES
Figure 1" Relations between variety and abundance (production) of aquatic life,
as organic pollution (discharged at mile 0) is carried down a stream. Time
and distance scales are only relative and will be found to differ in nearly every
case. After Bartsch aM Ingram. 1
-------
Biological Aspects of Natural Self Purification
e Bacterial count low
f Numbers of species high
g Numbers of organisms of each species
moderate or low
h Bottom free of sludge deposits
2 Characteristic biota includes a wide
variety of forms such as-
a A variety of algae and native higher
(vascular, or rooted) plants
b Caddis fly larvae (Trichoptera)
c Mayfly larvae (Ephemeroptera)
d Stonefly larvae (Plecoptera)
e Damselfly larvae (Zygoptera)
f Beetles (Coleoptera)
g Clams (Pelecypoda)
h Fish such as
- Minnows (Notropid types)
- Darters (Etheostomatidae)
- Millers thumb (Cottidae)
- Sunfishes and basses (Centrarchidae)
- Sauger, yellow perch, etc. (Percidae
- Others
3 Organisms characteristic of clean lakes,
estuaries, or oceanic shores might be
substituted for the above, and likewise
in the following sections. However, it
should be recognized that no single
habitat is as thoroughly understood in
this regard as the freshwater stream.
Ill POLLUTION
A With the introduction or organic pollution
(Figure 1, day 0), a succession of fairly
well organized events is initiated.
Important items to observe in interpreting
the pollutional significance of stream
organisms are the following
B Numbers of species present, they tend to
decrease with pollution.
C Numbers of individuals of each species
tends to increase with pollution.
D Ratios between types of organisms are
disturbed by pollution.
1 Clean water species intolerant of
organic pollution tend to become scarce
and unhealthy.
2 Animals with air breathing devices or
habits tend to increase in numbers.
3 Scavengers become dominant
4 Predators disappear
5 Higher plants, green algae, and most
diatoms tend to disappear.
6 Blue green algae often become
conspicious
E The importance of observations on any
single species is very slight.
IV THE ZONE OF RECENT POLLUTION
A The zone of recent pollution begins with
the act of pollution, the introduction of
excessive organic matter food for
microorganisms (Figure 1, day 0)
B There follows a period of physical mixing.
C Many animals and plants are smothered
or shaded out by the suspended material.
D With this enormous new supply of food
material, bacteria and other saprophytic
microorganisms begin to increase
rapidly.
-------
Biological Aspects of Natural Self Purification
E The elimination of intolerant predatory
animals allows the larger scavengers to
take full advantage of the situation.
F This explosive growth of organisms,
particularly fungi and bacteria, draws
heavily on the free dissolved oxygen for
respiration, and may eventually eliminate it.
G The number of types of organisms diminishes
but numbers of individuals of tolerant types
may increase.
H Zone of degeneration, or recent pollution,
can usually be characterized as follows:
1 General features:
a DO variable, 2 ppm to saturation
b BOD high
c Turbidity high
d Organic content high
e Bacterial count variable to high
f Number of species declines from
clean water zone
g Number of organisms per species
tends to increase
h Other- Slime may appear on bottom
2 Characteristic biota
a Fewer higher plants, but rank heavy
growth of those which persist
b Increase in tolerant green, and blue
green algae
c Midge larvae (Chironomidae) may
become extremely abundant
d Back swimmers (Corixidae) and water
boatmen (Notonectidae) often present
e Sludge worms (Tubificidae) common
• to abundant.
f Dragonflies (Anisoptera) often present
have unique tail breathing strainer
g Fish types, eg
- Fathead minnows (Pimephales
promelas)
- White sucker (Catostomus
commersonni )
- Bowfin (Amia calva)
- Carp (Cyprinus carpio)
V THE SEPTIC ZONE
A The exact location of the beginning of the
septic zone, if one occurs, varies with
season and other circumstances.
(Figure 1, day 1)
B Lack of free DO kills many microorganisms
and nearly all larger plants and animals,
again replenishing the mass of dead
organic material.
C Varieties of both macro and micro-
organisms and adjustable types (facultative)
that can live in the absence of free oxygen
(anaerobic) take over.
D These organisms continue to feed on their
bonanza of food (pollution) until it is
depleted.
E The numbers of types of organisms is now
at a minimum, numbers of individuals
may or may not be at a maximum.
F The septic zone, or zone of putrefaction
can usually be characterized as follows:
1 General features.
a Little or no DO during warm weather
b BOD high but decreasing
c Turbidity high, dark, odoriferous
d Organic content high but decreasing
-------
Biological Aspects of Natural Self Purification
e Bacterial count high
f Number of species very low
g Number of organisms may be extremely
high
h Other. Slime blanket and sludge
deposits usually present, oily
appearance on surface, rising gas
bubbles
2 Characteristic biota
a Blue green algae
b Mosquito larvae
c Rat-tailed maggots
d Sludge worms (Tubificidae and similar
forms). Small, red, segmented
(annelid) worms seem to be character-
istic of this zone in both fresh and
salt waters, the world around.
e Air breathing snails (Physa for
example)
f Fish types- None
3 Note Fortunately, all polluted waters
do not always degenerate to "septic"
conditions.
VI THE RECOVERY ZONE
A The septic zone gradually merges into the
recovery zone. (Figure 1, day 4)
B As the excessive food reserves diminish
so do the numbers of anaerobic organisms
and other pollution tolerant forms.
C As the excessive demand for oxygen
diminishes, free DO begins to appear and
likewise oxygen requiring (aerobic)
organisms.
D A^the suspended material is reduced and
available mineral materials increase due
to microbial action, algae begin to increase
often in great abundance.
E Photosynthesis by the algae releases more
oxygen, thus hastening recovery.
F Since algae require oxygen at all times
for respiration (like animals), heavy
concentrations of algae will deplete free
DO during the night when it is not being
replenished by photosynthesis.
G Consequently this zone is characterized
by extreme diurnal fluctuations in DO.
H With oxygen for respiration and algae, etc.
for food, general animal growth is resumed.
I The stream may now enter a period of
excessive productivity which lasts until
the accumulated energy (food) reserves
have been dissipated.
J Zone of recovery may usually be
characterized as follows-
1 General features
a DO 2 ppm to saturation
b BOD dropping
c Turbidity dropping, less color and
odor
d Organic content dropping
e Bacterial count dropping
f Numbers of species increasing
g Numbers of organisms per species
decreasing, (with the increase in
competition)
h Other: Less slime and sludge
2 Characteristic biota
a Blue green algae
b Tolerant green flagellates and other
algae
c Rooted higher plants in lower reaches
d Midge larve (Chironomids)
-------
Biological Aspects of Natural Self Purification
e Black fly larvae (Simulium)
f Giant water bugs (Belostoma spp.)
g Clams (Megalonais)
h Fish types
- Green sunfish (Lepomis cyanellus)
- Common sucker (Catostomus
commersonni)
- Flathead catfish (Pylodictis olivaris)
- Stone roller minnow (Campostoma
anomalum)
- Buffalo (Ictiobus cyprinellus)
3 Excessive production and extreme
variability often characterize middle and
lower recovery zones.
4 Unfortunately, many waters once polluted
never completely "recover". Re-
pollution is the rule in many areas so
that after the initial pollution, clear
out delineation of zones is not possible.
Characterization of these waters may
involve such parameters as productivity,
BOD, some "index" figure, or other
value not included here.
VII CLEAN WATER ZONE
A Clean water conditions again obtain when
productivity has returned to a normal,
relatively poor level, and a well balanced
varied flora and fauna are present.
(Figure 1, day "10") Conditions may
usually be characterized as follows-
B General features- similar to upstream
clean water except that it is now a larger
stream.
REFERENCES
1 Bartsch, A.F and Ingram, W.M
Stream Life and the Pollution Environ-
ment Public Works Publications,
July 1959, Vol. 90, No. 7, pp 104-110.*
2 Gaufin, A.R, and Tarzwell, C.M.
Aquatic invertebrates as indicators of
stream pollution. Reprint No 3141
from PHR. 67 (1) 57-64 1952.
3 Gaufin, A.R. and Tarzwell, C M.
Environmental changes in a polluted
stream during winter. Am. Midland
Naturalist. 54:68-88. 1955
4 Gaufin, A R. and Tarzwell, C.M.
Aquatic macro-invertebrate communities
as indicators of organic pollution in
Lytle Creek. Sewage and Ind Wastes
28-906-24 1956
5 Hynes, H.B.N. The Biology of Polluted
Waters Liverpool Univ Press,
pp. 202. 1963
6 Katz, M. and Gaufin, A.R The effects
of sewage pollution on the fish population
of a midwestern stream. Trans. Am.
Fisheries Soc. 82-156-65 1952. *
7 Reish, D. J The Relationship of the
Polychaetous Annelid Capitella capitata
(Fabricius) to Waste Discharges of
Biological Origin In: Biol. Prob
Water Pol. - Trans 1959 Seminar.
Robert A. Taft Sanitary Engineering
Center, USPHS, Cincinnati, OH.
pp. 195-200.
8 Biology of Water Pollution FWQA Pub.
CWA-3 (references with an asterisk
are reprinted in this publication. 1967
C Characteristic biota- similar to upstream
clean water fauna and flora except that
species include those indigenous to a
larger stream.
This outline was prepared by H. W. Jackson,
Chief Biologist, National Training Center,
DTTB, MDS, WPO, EPA, Cincinnati,
OH 45268.
-------
ECOLOGY OF WASTE STABILIZATION PROCESSES
I INTRODUCTION
Living organisms will live where they can live.
This holds for treatment plant environments
just as it does for streams, impoundments,
oceans, dry or wet lands.
A Each species has certain limits or toler-
ances, growth, feeding habits and other
characteristics that determine its favored
habitat.
B The presence of certain organisms with
well defined characteristics in a viable
condition and in significant numbers also
provides some inference with respect to
the habitat.
C The indicator organism concept has certain
pitfalls. It is not sufficient to base an
opinion upon one or more critters which
may have been there as a result of gas
liquid or solid transport. It is necessary
to observe growth patterns, associated
organisms, environmental conditions, and
nutritional characteristics to provide
information on environmental acceptability.
D Organisms characteristic of wastewater
treatment commonly are those found in
nature under low DO conditions. Perform-
ance characteristics are related to
certain organism progressions and assoc-
iations that are influenced by food to
organism ratios and pertinent conditions.
One single species is unlikely to perform
all of the functions expected during waste
treatment. Many associated organisms
compete in an ecological system for a
favored position. The combination includes
synergistic, antagonistic, competitive,
predative, and other relationships that may
favor predominance of one group for a time
and other groups under other conditions.
E It is the responsibility of the treatment
plant control team to manage conditions
of treatment to favor the best attainable
performance during each hour of the day
each day of the year. This outline con-
siders certain biological characteristics
and their implications with respect to
treatment performance.
H TREATMENT PLANT ORGANISMS
Wastewater is characterized by overfertili-
zation from the standpoint of nutritional
elements, by varying amounts of items that
may not enter the metabolic pattern but have
some effect upon it, such as silt, and by
materials that will interfere with metabolic
patterns. Components vary in availability
from those that are readily acceptable to
those that persist for long periods of time
Each item has some effect upon the organism
response to the mixture.
A Slime forming organisms including certain
bacteria, fungi, yeasts, protista monera
and alga tend to grow rapidly on dissolved
nutrients under favorable conditions. These
grow rapidly enough to dominate the overall
population during early stages of growth.
There may be tremendous numbers of
relatively few species until available
nutrients have been converted to cell mass
or other limiting factors check the pop-
ulation explosion.
B Abundant slime growth favors production
of predator organisms such as amoeba or
flagellates. These feed upon preformed
cell mass. Amoebas tend to flow around
particulate materials; flagellates also are
relatively inefficient food gatherers. They
tend to become numerous when the nutrient
level is high. They are likely to be assoc-
iated with floculated masses where food
is more abundant.
C Ciliated organisms are more efficient
food gatherers because they have the
ability to move more readily and may
set up currents in the water to bring food
to them for ingestion. Stalked ciliates
are implicated with well stabilized effluents
because they are capable of sweeping the
fine particulates from the water between
floe masses while their residues tend to
become associated with the floe.
D Larger organisms tend to become establish-
ed later and serve as scavengers. These
include Oligochaetes (worms), Chironomids
(bloodworms and insect larvae), Isopods
(sow bugs and Crustacea), Rotifera and
others.
PC. 19.10.69
-------
Ecology of Waste Stabilization Processes
in TREATMENT OPERATIONAL CONTROL
An established treatment plant is likely to
contain representative organisms from all
groups of tolerant species. Trickling filters,
activated sludge, or ponds tend to retain
previously developed organisms in large
numbers relative to the incoming feed. The
number and variety available determine the
nature, degree and time required for partial
oxidation and conversion of pollutants from
liquid to solid concentrates.
A Proliferation of slime forming organisms
characterize the new unit because they grow
rapidly on soluble nutrients. Predators
and scavengers may start growing as soon
as cell mass particulates appear but growth
rate is slower and numbers and mass lag
as compared with slime organisms. As
slime growth slows due to conversion of
soluble nutrients to cell mass, the slime
formers tend to associate as agglomerates
or clumps promoting floculation and liquid
solid separation.
B Overfeeding an established unit encourages
rapid growth of slime organisms as individ-
ual cells rather than as flocculated masses.
This results in certain characteristics
resembling those of a young, rapidly
growing system.
C Toxic feeds or unfavorable conditions
materially reduce the population of exposed
sensitive organisms. The net effect is a
population selection requiring rapid regrowth
to reestablish desired operating character-
istics. The system assumes new growth
characteristics to a degree depending upon
the fraction remaining after the toxic effect
has been relieved by dilution, degradation,
sorption, or other means.
D Treatment units are characterized by
changes in response to feed sequence,
load ratio, and physical or chemical
conditions. Response to accute toxicity
may be immediately apparent. Chronic
overloading or mild toxicity may not be
apparent for several days. It may be
expected that it will require 1 to 3 weeks
to restore effective performance after any
major upset. Performance criteria may
not indicate a smooth progression toward
improved operation.
E Observations of the growth characteristics
and populations do not provide quantitative
information, but they do indicate trends
and stages of development that are useful
to identify problems. It is not possible to
identify most slime organisms by direct
observation. It is possible to recognize
growth and flocculation characteristics.
Certain larger organisms are recognizable
and are useful as indicator organisms to
suggest past or subsequent developments.
IV ILLUSTRATIONS OF ECOLOGICAL
SIGNIFICANCE
A The first group represents initial devel-
opment of non-flocculent growth. Single
celled and filamentous growth are shown.
Rapid growth shows little evidence of
flocculation that is necessary to produce
a stable, clear effluent.
B The next group of slides indicate develop-
ment of floe forming tendencies from
filamentous or non-filamentous growth.
Clarification and compaction characteris-
tics are relatable to the nature and density
of floe masses.
C Organisms likely to be associated with
more stabilized sludges are shown in the
third group. Scavengers essentially con-
sist of a large alimentary canal with
accessories.
D The last two slides illustrate changes in
appearance after a toxic load. Scavengers,
ciliates, etc. have been inactivated. New
growth at the edge of the floe masses are
not apparent. Physical structure indicates
dispersed residue rather than agglomera-
tion tendencies. The floe probably contains
living organisms protected by the surround-
ing organic material, but only time and
regrowth will reestablish a working floe
with good stabilization and clarification
tendencies.
A CK NOW LE DGEME NTS
This outline contains significant materials from
previous outlines by H. W. Jackson and R M.
Sinclair. Slide illustrations were provided by
Dr. Jackson.
This outline was prepared by F J Ludzack.
Chemist, National Training Center, WPO,
EPA, Cincinnati, OH 45268.
-------
EFFECTS OF POLLUTION ON FISH
the bodies of fishes by various routes.
We are generally not aware of their
presence unless they:
1 Cause an observable effect on the fish,
2 Cause an effect on man by imparting
off-taste or odor to fish flesh,
3 Are sought for and detected, e.g.,
radioactive substances, DDT, mercury.
I INTRODUCTION
A By what means do pollutants exert their
effects'
B What is the relationship between water
quality and water use by fishes'
C What is the reaction of fishes to domestic
sewage'
D Is there any noticeable change in species
composition of the population following
pollution'
E Is there any genetic or environmental
selection in favor of pollution resistant
strains'
F Nearly any pollutant, given sufficient
concentration and time, can kill as a
direct toxicant. We are primarily con-
cerned here with sub-lethal or chronic
levels of pollutants. (Acute toxic levels
and physiological mechanisms are
treated elsewhere.)
II MECHANISMS OF DETRIMENTAL ACTION
A In^rt silt may
1 Clog gills and smother eggs and fry,
2 Blind sight feeders and eliminate
hiding places,
3 Smother food organisms,
4 Redace oxygenation by smothering
algae.
B Irritants may
1 Act as repellents,
2 Cause excessive mucous secretion and
upset osmotic balance.
C Sub-lethal quantities of a host of environ-
mental materials are constantly penetrating
We can only speculate as to their
undetected effects.
Ill ENVIRONMENTAL RELATIONSHIPS
BETWEEN WATER QUALITY AND WATER
USE BY FISHES
A Freshwater fishes sometimes spend their
entire lives in a single body of water.
Pollution of that body of water therefore
impinges on them at every stage of their
life cycle, and at every point in their
various ecological relationships; such as,
seeking food or escaping enemies.
B Migratory fisnes on the other hand feed and
grow up m one body of water (the ocean
for anadromous species, fresh water for
the catadronous eels), then travel a migra-
tion route (usually a river) to another body
of water whsre they breed.
Pollution at either end of the route, or a
pollution block along the migration route,
may eliminate the species.
C What will affect one species adversely may
be favorable for another.
1 Cold water species, such as various
trodts, might be killed or eliminated by
warmed water from a power plant which
would in turn permit the survival of warm
water species, such as certain basses,
sunfishes, etc.
2 Benthic species (such as catfish, sculpins,
or suckers, whicn live near the bottom)
might be eliminated by a smothering
BI. FI. 11a. 5. 71
-------
Effects of Pollution on Fish
blanket of inert material which would
not affect limnetic species inhabiting
the open water areas {such as white
and yellow bass, gizzard shad, or
walleye). The limnetic specie3 on the
other hand might be inhibited by a dense
turbidity which would hide their prey,
suppress the growth of nutritious plank-
ton, or clog their gills, this in turn
being relatively harmless to the benthic
group.
3 Likewise the shoreline hugging littoral
forms (like pumpkinseed or bluegills)
and profundal species (such as lake
trout) might respond selectively to such
factors as temperature or transparency.
IV RESPONSE OF FISHES TO SEWAGE AND
SIMILAR WASTES
A These wastes in general are not toxic in
themselves, but exert their effects on
fishes directly.
B Oxygen Depletion
1 May lead to death at various stages m
life history depending on circumstances.
2 May lower resistance to disease or
increase sensitivity to intoxication.
3 May reduce ability to capture food or
swim against current.
C May smother or kill normal food sources.
D May increase normal fish production
through eutrophication.
E Usually changes normal population balance
by driving out predatory types and en-
couraging scavengers.
F Reported to cause osteological and other
pathological manifestations, such as the
knothead condition of carps in the Illinois
River.
V NATURAL SELECTION AND
ACCLIMATIZATION TO POLLUTION
A Known biological mechanisms for selective
breeding of pollution resistant strains
operate in nature among fishes as among
other organisms.
1 Studies of population genetics indicate
that after some finite number of genera-
tions of population stress (e.g.; exposure
to a given pollutant), permanent heritable
resistance may be expected to develop.
2 If the environmental stre3s (or pollutant)
is removed prior to the time that
permanent resistance is developed in
the population, reversion to the non-
resistant condition may occur within a
relatively few generations.
3 Habitats harboring populations under
stress in this manner are often marked
with the dead bodies of the unsuccessful
individuals.
B Individual organisms on the other hand can
over a period of time (less than one life
cycle) develop a limited ability to tolerate
different conditions, e.g. ; pollutants:
1 With reference to all categories of
pollutants both relatively facultative
and obligate species are encountered
(e.g.; eurynaline vs. stenohaline,
eurythermal vs. stenothermal).
2 Th's temporary somatic acclimatization
is not heritable.
C A given single-species collection or sample
of living fishes may therefore represent
one or more types of pollution resistance:
1 A sample of an original population which
has been acclimated to a given stress in
toto.
2 A sample of the surviving portion of
an original population, which "ias been
"selected" by the ability to endure the
stress. The dead fish in a partial fish
kill are that portion of the original
population unable to endure the stress.
-------
Effects of Pollution on Fish
3 A sample of a sub-population of the
original species in question which has
in tot o over a period of several genera-
tions developed a heritable stress
resistance.
D Any given multi- species field collection
will normally contain species illustrative
of one or more of the conditions outlined
above.
REFERENCES:
1 California, State of. Water Quality
Criteria, 2nd ed. Resources Agency
of California, State Water Quality Con-
trol Board Publication No. 3-A. 1963.
2 Foross, S.A., and Richardson, R.E. Some
Recent Changes in Illinois River Biology.
Bull. 111. Nat. His". Sur., 13(6) 1919.
VI POPULATION COMPOSITION RESPONSES
TO POLLUTION
A Sewage pollution generally results in a
reduction in the predatory types and their
replacement by scavengers. Regions of
severe oxygen depletion may be devoid of
fish, or inhabited only by rough fish such
as gar or carp. The general concept of a
reduction of variety coupled with in in-
crease m abundance in certain regions is
as valid for fishes as for other groups.
B Population responses to toxic pollution are
unpredictable except that reduction in
variety is again almost sure to result.
3 Jones, J.R. Erichsen, Fish and River
Pollution. Butterworth's London,
pp. 203. (1964)
4 Katz, Max, and Gaufin, A.R. The Effects
of Sewage Pollution on the Fish Popula-
tion of a Midwestern Stream. Midwestern
Stream. Trans. Am. Fish. Soc. 82-156-
165. 1952.
5 Moore, Emmeline. S1 ream Pollution and
Its Effects on Fish Life. Sewage Works
Journal. 4:159. 1932.
6 Naegele, John. A. Head, Dept. of En/iron-
mental Sciences, Univ. of Mass.,
Waltham Field Station, Waltham, Mass.
Personal Communication. 1965.
7 Trautman, M. B. The General Effects of
Pollution m Ohio Fish Life. Trans. Am.
Fish. Soc. 63 69-72. 1933.
8 Mills, Harlow B.; S^arrett, William C.,
and Bellrose, Frank C. Mai's Effect
on the Fish and Wildlife of the Illinois
River. 111. Nat. Hist. Surv. Biol.
Notes No, 57. 24pp. 1966.
«
9 Warren, Charles E. Biology and Water
Pollution Control. W. B. Saunders
Co. 434 pp. 1971.
Th\s outline was prepared 'jy H. W. Jackson
Chief Biologist, National Training Center,
EPA, Cincinnati, OH 452 68.
-------
THE INTERPRETATION OF BIOLOGICAL DATA
WITH REFERENCE TO WATER QUALITY
I INTRODUCTION
Sanitary engineers like to have data
presented to them in a readily assimilable
form and some of them seem a little
impatient with biologists who appear unable
to provide definite quantitative criteria
applicable to all kinds of water conditions.
I think the feeling tends to be that this is
the fault of biologists, and if they would
only pull themselves out of the scientific
stone-age all would be well. I will try to
explain here why I believe that biological
data can never be absolute nor mterpret-
able without a certain amount of expertise.
In this respect biologists resemble medical
men who make their diagnoses against a
complex background of detailed knowledge.
Anyone can diagnose an open wound but it
takes a doctor to identify an obscure
diseasej and although he can explain how
he does it he cannot pass on his knowledge
in that one explanation. Similarly, one
does not need an expert to recognize gross
organic pollution, but only a biologist can
interpret more subtle biological conditions
in a water body, and here again he can
explain how he does it, but that does not
make his hearer a biologist. Beck (1957)
said something similar at a previous
symposium in Cincinnati in 1956.
II THE COMPLEXITY OF BIOLOGICAL
REACTIONS TO WATER CONDITIONS
A Complexity of the Aquatic Habitat
The aquatic habitat is complex and
consists not only of water but of the
substrata beneath it, which may be
only indirectly influenced by the quality
of the water. Moreover, in biological
terms, water quality includes such
features as rate of flow and tempera-
ture regime, which are not considered
of direct importance by the chemist.
To many animals and plants, maximum
summer temperature or maximum
rate of flow is just as important as
minimum oxygen tension. The result
is that inland waters provide an
enormous array of different com-
binations of conditions, each of which
has its own community of plants and
animals, and the variety of species
involved is very great. Thu3, for
example, Germany has about 6000
species of aquatic animals (lilies 1961a)
and probably at least as many species
of plants. Yet Europe has a rather
restricted fauna because of the
Pleistocene ice age, in most other
parts of the world the flora and fauna
are even richer.
B Distribution of Species and Environ-
mental Factors
We know something about the way in
which species are distributed in the
various habitats, especially in the
relatively much studied continent of
Europe, but we have, as yet, little
idea as to what factors or combination
of factors actually control the individual
species.
1 Important ecological factors
Thus, it is possible to list the
groups of organisms that occur in
swift stony upland rivers
(rhithron in the sense of lilies,
1961b) and to contrast them with
those of the lower sluggish reaches
(potamon). Similarly we know,
more or less, the different floras
and faunas we can expect in
infertile (oligotrophic) and fertile
(eutrophic) lakes. We are, however,
much less informed as to just what
ecological factors cause these
differences. We know they include
temperature and its yearly
BI.EN. Id. 3.71
-------
The Interpretation of Biological Data with Reference to Water Quality
amplitude; oxygen, particularly at
minimal levels; plant nutrients,
such as nitrate, phosphate, silica,
and bicarbonate; other ions in
solution, including calcium, chloride,
and possibly hydrogen; dissolved
organic matter, which is necessary
for'some bacteria and fungi and
probably for some algae, the nature
of the substratum; and current.
2 Complexity of interacting factors
We also know these factors can
interact in a complex manner and
that their action on any particular
organism can be indirect through
other members of the biota.
a Induced periphyton growths
Heavy growths of encrusting
algae induced by large amounts
of plant nutrients, or of
bacteria induced by ample
supplies of organic matter,
can eliminate or decimate
populations of lithophile insects
by simple mechanical inter-
ference. But the change does
not stop there1 the growths
themselves provide habitats
for the animals, such as
Chironomidae and Naidid worms,
which could not otherwise live
on the stones.
b Oxygen levels and depositing
substrates
If oxygen conditions over a
muddy bottom reach levels
just low enough to be intolerable
to leeches, tubificid worms,
which the leeches normally
hold in check, are able to build
up to enormous numbers
especially as some of their
competitors (e.g. Chironomus)
are also eliminated.
c Oxygen levels and non muddy
substrates
One then finds the typical
outburst of sludge worms, so
often cited as indicators of
pollution. This does not
happen if the same oxygen
tension occurs over sand or
rock, however, as these are
not suitable substrata for the
worms. Many such examples
could be given, but they would
only be ones we understand;
there must be a far greater
number about which we know
nothing.
d One must conclude, therefore,
that quite simple chemical
changes can produce far-
reaching biological effects,
that we only understand a
small proportion of them; and
that they are not always the
same.
3 Classic examples
This seems like a note of despair,
however, if water quality deviates
too far from normal, the effects
are immediately apparent. Thus,
poisonous substances eliminate
many species and may leave no
animals {Hynes 1960); excessive
quantities of salt remove all
leeches, amphipods, and most
insects and leave a fauna con-
sisting largely of Chironomidae,
caddis worms, and oligochaetes
(Albrecht 1954) and excessive
amounts of dissolved organic
matter give rise to carpets of
sewage fungus, which never occur
naturally. Here no great biologi-
cal expertise is needed, and there
is little difficulty in the
communication of results. It is
when effects are slighter and more
subtle that biological fmdings
-------
The Interpretation of Biological Data with Reference to Water Quality
become difficult to transmit
intelligibly to other disciplines.
Ill THE PROBLEMS IN PRESENTATION
OF BIOLOGICAL RESULTS
Because of these difficulties various
attempts have been made to simplify the
presentation of biological findings, but to
my mind none of them is very successful
because of the complexity of the subject.
Early attempts at systematization developed
almost independently on the two sides of
the Atlantic, although they had some
similarities.
A Early Studies in the United States
(Richardson and the Illinois River)
In America, there was a simple division
into zones of pollution, e.g. degradation,
septic, and recovery, which were
characterized in broad general terms.
This simple, textbook approach is
summarized by Whipple et al. (1947),
and serves fairly well for categorizing
gross organic pollution such as has been
mentioned above. It was, however,
soon found by Richardson (1929) during
his classical studies on the Illinois
River that typical "indicators" of foul
conditions, such as Tubificidae and
Chironomus, were not always present
where they would be expected to occur.
This was an early indication that it is
not the water quality itself that provides
suitable conditions for "pollution faunas, "
but other, usually associated, conditions -
in this instance deposits of rich organic
mud. Such conditions may, in fact, be
present in places where water quality
in no way resembles pollution, e. g.,
upstream of weirs in trout streams
where autumn leaves accumulate and
decay and cause the development of
biota typical of organically polluted
water. Samples must therefore be
judged against a background of biological
knowledge. Richardson was fully aware
of this and was in no doubt about the
condition of the Illinois River even in
places where his samples showed few
or no pollution indicators.
B The European Saprobic System
In Europe, the initial stress was
primarily on microorganisms and
results were first codified in the
early years of the century by
Kolkwitz and Marsson. In this
"Saprobiensystem, " zones of organic
pollution similar to those described
by the American workers were defined
and organisms were listed as charac-
teristic of one or more zones;
TABLE 1
SAPROBIENSYSTEM - A European system
of classifying organisms according to their
response to the organic pollution in slow
moving streams. (22)
Alpha-Mesosaprobic Zone - Area of
active decomposition, partly aerobic,
partly anaerobic, in a stream heavily
polluted with organic wastes.
Beta-Mesosaprobic Zone - That reach
of stream that is moderately polluted
with organic wastes.
Ohgosaprobic Zone - That reach of a
stream that is slightly polluted with
organic wastes and contains the
mineralized products of self-
purification from organic pollution,
but with none of the organic pollutants
remaining.
Polysaprobic Zone - That area of a
grossly polluted stream which contains
the complex organic wastes that are
decomposing primarily by anaerobic
processes.
A recent exposition of this list is
given by Kolkwitz (1950). It was then
claimed that with a list of the species
occurring at a particular point it was
possible to allocate it to a saprobic
zone. This system early met with
criticism for several reasons. First,
-------
The Interpretation of Biological Data with Reference to Water Quality
TABLE 2
SAFROBICITY LEVELS ACCORDING TO THE TROPHIC
STRUCTURE OF THE COMMUNITIES OF ORGANISMS
Saprobicity Level Structure of the Communities of Organisms
I |3-okgosaprobic Balanced relationship between producers, consumers
and destroyers, the communities of organisms are
poor in individuals but there is a moderate variety of
species, small biomass and low bioactivity.
II o-oligosaprobic Balanced relationship between producers, consumers
and destroyers, communities of organisms are rich in
individuals and species with a large biomass and high
bioactivity.
Ill /3-mesosaprobic Substantially balanced relationship between producers,
consumers and destroyers, a relative increase in the
abundance of destroyers and, accordingly, of the con-
sumers living off them, communities of organisms are
rich in individuals and species with a large biomass and
high bioactivity.
IV a-mesosaprobic Producers decline as compared with an increase in
consumers and destroyers, mixotrophic and amphitrophic
forms predominate among the producers, communities of
organisms rich in individuals but poor in species with a
large biomass and extremely high bioactivity; still only
few species of macro-organisms; mass development of
bacteria and bacteria-eating ciliates.
V /3-polysaprobic Producers drastically decline, communities of organisms
are extremely rich in individuals but poor m species with
a large biomass and high bioactivity; macrofauna represented
only by a few species of tubificids and chironomids; as in
IV these are in great abundance; mass development of
bacteria and bacteria-eating ciliates.
VI of-polysaprobic Producers are absent; the total biomass is formed
practically solely by anaerobic bacteria and fungi;
macro-organisms are absent, flagellates outnumber
ciliates amongst the protozoa.
Saprobicity - "Within the bioactivity of a body of water, Saprobicity is the sum
total of all those metabolic processes which are the antithesis of
primary production. It is therefore the sum total of all those
processes which are accompanied by a loss of potential energy. "
Part I, Prague Convention.
-------
The Interpretation of Biological Data with Reference to Water Quality
all the organisms listed occurred in
natural habitats--they were not evolved
in polluted water--and there was much
doubt as to the placing of many of the
species in the lists. The system, how-
ever, did serve to codify ecological
knowledge about a long list of species
along an extended trophic scale. Its
weaknesses appeared to be merely due
to lack of knowledge; such a rigid
system took far too little account of the
complexity of the reaction of organisms
to their habitats. For instance, many
organisms can be found, albeit rarely,
in a wide range of conditions and others
may occur in restricted zones for
reasons that have nothing to do with
water quality. We often do not know if
organisms confined to clean headwaters
are kept there by high oxygen content,
low summer temperatures, or inability
to compete with other species under
other conditions. In the swift waters of
Switzerland the system broke down in
that some organisms appeared in more
polluted zones than their position in the
lists would indicate. Presumably here
the controlling factor was oxygen, which
was relatively plentiful in turbulent cold
water. In a recent series of experiments,
Zimmerman (1962) has proven that
current alone has a great influence on
the biota, and identically polluted water
flowing at different speeds produces
biotic communities characteristic of
different saprobic levels. He finds this
surprising, but to me it seems an
expected result, for the reasons given
above.
C Recent Advances in the Saprobic System
1 Perhaps Zimmerman's surprise
reflects the deeply rooted entrench-
ment of the Saprobiensystem in
Central Europe. Despite its obvious
shortcomings it has been revised
and extended. Liebmann (1951)
introduced the concept of consider-
ing number as well as occurrence
and very rightly pointed out that the
community of organisms is what
matters rather than mere species
lists. But he did not stress the
importance of extrinsic factors,
such as current, nor that the
system can only apply to organic
pollution and that different types
of organic pollution differ m their
effects, e.g., carbohydrate solu-
tions from paper works produce
different results from those of
sewage, as they contain little
nitrogen and very different sus-
pended solids. Other workers
(Sladecek 1961 and references
therein) have subdivided the more
polluted zones, which now, instead
of being merely descriptive, are
considered to represent definite
ranges of oxygen content, BOD,
sulfide, and even E. coli populations.
Every water chemist knows that
BOD and oxygen content are not
directly related and to assume that
either should be more than vaguely
related to the complexities of
biological reactions seems to me
to indicate a fundamental lack of
ecological understanding. I also
think it is damaging to the hope of
mutual understanding between the
various disciplines concerned with
water quality to give the impression
that one can expect to find a close
and rigid relationship between
water quality measurements as
assessed by different sets of
parameters. Inevitably these
relationships vary with local con-
ditions, what applies in a sluggish
river in summer will certainly not
apply to a mountain stream or even
to the same river in the winter.
Correlation of data, even within
one discipline, needs understanding,
knowledge, and judgment.
2 Caspers and Schulz (1960) showed
that the failure of the system to
distinguish between waters that are
naturally productive and those
artifically enriched can lead to
absurd results. They studied a
canal in Hamburg, which because
of its urban situation can only be
regarded as grossly polluted.
Yet it develops a rich plankton.
-------
The Interpretation of Biological Data with Reference to Water Quality
the composition of which, according
to the system, shows it to be
virtually clean.
D Numerical Application of the Saprobic
System
Once the Saprobiensystem was accepted
it was logical to attempt to reduce its
findings to simple figures or graphs for
presentation of results. Several such
methods were developed, which are
described by Tumpling (I960), who also
gives the original references. In all
these methods, the abundance of each
species is recorded on some sort of
logarithmic scale (e.g. 1 for present,
3 for frequent, 5 for common, etc.)
The sums of these abundances in each
saprobic level are plotted on graphs,
the two most polluted zones showing as
negative and others as positive. Or, the
various saprobic levels are given
numerical values [1 for oligosaprobic
(clean), 2 for j3-mesosaprobic, etc.]
and the rating for each species is
multiplied by its abundance number.
The sum of all these products divided
by the sum of all the frequencies gives
a "saprobic index" for the locality.
Clearly the higher this number, the
worse the water quality in terms of
organic pollution. In a similar way the
so-called "relative Belastung" (relative
load) is calculated by expressing the
sums of all the abundances of organisms
characteristic of the two most-polluted
zones as a percentage of the sum of all
abundances. Then 100 percent is
completely polluted water, and clean
localities will give a low number.
authors in the assignment of species to
the different levels. Therefore, one
gains a number or a figure that looks
precise and is easily understood, but
it is based on very dubious foundations.
F Comparative North American Systems
Similar systems are indigenous to
North America, but were independently
evolved.
1 Wurtz (1955) and Wurtz and Dolan
(1960) describe a system whereby
animals are divided into sensitive-
to-pollution and non-sensitive
(others are ignored), and also into
burrowing, sessile, and foraging
species (six classes).
BSFP BSFP BSFP BSFP BSFP BSFP BSFP BSFP BSFP
;40t
0000
10
M-l 16
i 60- -
80--
100 x
M*7 M-0
M-6 0/28*8 El/28/48
14 M-5 9/6*8 RECOVW OEGRAD
M-3 U.A 8/2W8 RECOVERY
8/2ft*8 &>Z€Aa RECOVER* ,
DEGRADr-^SEPTIC.
—FLOW
A-L
3 8/26*8
SEPTIC
Figure 1. Histograms, based on selected organisms, iflustraling stream
reaches of clean, degradation, septic, and recovery conditions [after
Wurfi] <22)
E Weaknesses of the Saprobic System
There are various elaborations of these
methods, such as sharing of species
between zones and taking account of
changes in base-line as one passes
downstream. None of them, however,
eliminates the basic weaknesses of the
system nor the fact that, as Caspers
and Schulz (1960) point out, there is
little agreement between the various
Numbers of these species rep-
resented are plotted for each station
as six histograms on the basis of
percentage of total number of
species. If the constitution of the
fauna from control stations or from
similar localities is known, it is
possible to express numerically
"biological depression" (i.e.,
percentage reduction in total
-------
The Interpretation of Biological Data with Reference to Water Quality
number of species), "biological
distortion (changes in pro-
portions of tolerant and non-tolerant
species), and "biological skewness"
(changes in the ratios of the three
habitat classes). Such results must,
of course, be evaluated, and the
definition of tolerance is quite
subjective; but the method has the
advantages of simplicity and depend-
ence on control data. Like the
Saprobiensystem, however, it can
have no universal validity. It also
suffers from the fact that it takes
no account of numbers, a single
specimen, which may be there by
accident, carries as much weight
as a dense population.
Patrick (J.949) developed a similar
system in which several clean
stations on the water body being
investigated are chosen, and the
average number of species is deter-
mined occurring in each of seven
groups of taxa chosen because of
their supposed reaction to pollution.
These are then plotted as seven
columns of equal height, and data
from other stations are plotted on
the same scale, it is assumed that
stations differing markedly from the
controls will show biological
imbalance in that the columns will be
of very unequal heights. Number is
indicated by double width in any
column containing species with an
unusual number of individuals.
I have already questioned the use-
fulness of this method of presentation
(Hynes I960), and doubt whether it
gives any more readily assimilable
data than simple tabulation, it does
however, introduce the concept of
ecological imbalance.
HEALTHY
POLLUUO
e
1
UlJ
I lit IV v
SEMI-HEALTHY
ii 111 iv v yi m
VERY POLLUTED
Figure 2. Histograms, based on telecied organisms, illustrating healthy,
semi-healthy, polluted, ond very polluted stations in Conestoga
Basin, Pa | after Patrick J (22)
TABLE 3 —CfaisiReation of Group*
of Organisms Shown in Figuro2
I Blue-green algae, green algae of the genera S'rgeocfonrum, Spi
rogyro, and Tnbonema, the bdclloid rotifers plui Cepha/odef/o
megofocep/iofa and Proa/es decipienj
II Oligochaetes, leeches, and pulmonale snails
III Protozoa
IV Diatoms, red algae, ond ' most of the green algae"
V All rotifers not included in Group I, clams, gill breathing mails,
and tnctadid flatworms
VI All insects and Crustacea
VII All fish
Beak (1964), another author,
recognized the need for a concise
expression of pollution based on
biological information. Toward
this end, he developed a method of
biological scoring which is based on
the frequency of occurrence of
certain macroscopic invertebrates
obtained from 6 years of study on
one river. It will be noted that the
Biological Score is a modification
and expansion of Beck's Biotic
Index.
-------
The Interpretation of Biological Data with Reference to Water Quality
The indicator organisms are
divided into three categories:
Group I contains the pollution -
tolerant species, Group II comprises
those species which are facultative
with respect to pollution, and
Group III contains the pollution -
intolerant forms. Each group is
assigned a weighted score that can
be allotted to field samples on the
following basis-
a Normal complement of Group III
scores 3 points.
b Normal complement of Group II
scores 2 points.
c Normal complement of Group I
scores 1 point.
The scores are additive, thus an
unpolluted stream will have a
Biological Score of 6. If only
pollution-tolerant forms are found,
the score will be 1. If no organisms
are found, the score will be zero.
Furthermore, a score of, 1 or 2
points could be allotted to Group III
when less than the normal com-
plement is present Group II could
be treated in a similar manner
This scoring device correlated well
witn the biological oxygen demand,
dissolved oxygen, and solids content
of the receiving water Beak also
related his scoring device to the
fisheries potential. This relation-
ship is shown in Table 4]
TABLE 4
Tentativk Relationship of the Biologicu. Score to the Fisheries
Potential (after Beak, !9b 1) (30)
It has long been known that
ecologically severe habitats con-
tain fewer species than normal
habitats and that the few species
that can survive the severe con-
ditions are often very abundant as
they lack competitors Examples
of this are the countless millions
of Artemia and Ephydra in saline
lakes and the Tubifex tubifex in
foul mud This idea has often been
expressed in terms of diversity,
which is some measure of numbers
of species divided by number of
specimens collected Clearly,
such a parameter is larger the
greater the diversity, and hence
the normality of the habitat
Unfortunately, though as the
number of species in any habitat
is fixed, it also decreases as
sample size increases so no index
of diversity has any absolute value
(Hairston 1959) If a definite
sample size is fixed, however, in
respect to numbers of organisms
identified, it is possible to arrive
at a constant index
8 20
10
r
/1 \i.
Pollution status
Biotic index
Fisheries potential
0 10 ,0
Miles from source
Figure 3 Zooplankton species diversity
per thousand individuals encountered in
marine systems affected by waste waters
from petrochemical industrial wastes
The vertical lines indicate seasonal
variations. (30)
Unpolluted
Slight to moderate pollution
Moderate pollution
Moderate to heavy pollution
Heavy pollution
Severe pollution, usually Iomc
5 or 4
3
2
1
0
All normal fisheries for tjpe of
water well developed
Most sensitive fish species re-
duced in numbers or mi-sing
O.'ih coarse fisherici maintained
If fish present, only those nitli
high toleration of pollution
Very little, if atij, fishery
No fish
-------
The Interpretation of Biological Data with Reference to Water Quality
5 Patrick et al. (1954) in effect used
this concept in a study of diatom
species growing on slides suspended
in water for fixed periods They
identified 8000 specimens per
sample and plotted the results as
number of species per interval
against number of specimens per
species on a logarithmic scale
This method of plotting gives a
truncated normal curve for a wide
variety of biotic communities
In an ordinarily diverse habitat the
mode is high and the curve short,
l e , many species occur in small
numbers and none is very abundant
In a severe habitat the mode is low
and the curve long, i e .there are
few rare species and a few with
large numbers This, again, seems
to me to be an elaborate way of
presenting data and to involve a lot
of unnecessary arithmetic
1, Ncnpolljted stream
B. Polluted sttea-n
1-2 2-4 12-64(1-12., 1?3- ,5 S'.- JC74- ?fil 40,5-
?58 512 'W r/i WS1 *192
Number of , r: srscrss
Figure 4. A graphic comparison of diatom
communities from two different environ-
ments. (After Patrick et al , 1954) (30)
6 Diversity indices vs tabulated data
Allanson (1961) has applied this
method to the invertebrate faunas
of streams in South Africa and has
shown, as has Patrick for diatoms,
that the log normal curve is flatter
and longer for polluted stations,
the difference, however, is not so
apparent that it does not need
exposition Here, again, I would
suggest that tabulated data are just
as informative Indeed I would go
further and say that tabulated data
are essential in the present state
of our knowledge We are learning
as we go along and if the details of
the basic findings are concealed by
some sort of arithmetical manip-
ulation they cannot be re-interpreted
in the light of later knowledge, nor
are they preserved in the store of
human knowledge This point
becomes particularly clear when
one examines some of the early
studies that include tables
Butcher (1946) requotes a con-
siderable amount of data he
collected from studies of various
English rivers during the thirties,
they are not only clear and easy to
follow, but they are also informative
about the generalities of pollution
in a way that data quoted only
within the confines of some
particular system are not
7 Examples of tabulated data(Table 5)
Simple tabulation of biological data
in relation to water quality, either
in terms of number of organisms,
percentage composition of the biota,
some arbitrary abundance scale,
or as histograms, has been
effectively practiced in many parts
of the world in America (Gaufin
and Tarzwell 1952, Gaufin 1958),
Africa (Harrison 1958 and 1960,
Hynes and Williams 1962), Europe
(Albrecht 1954, Kaiser 1951,
Hynes 1961, Hynes and Roberts
1962), and New Zealand (Hirsch
1958) to cite a few These tabu-
lated data are easy to follow, are
informative to the expert reader,
and conceal no facts. Although the
non-biologist may find them tedious,
he need only read the explanatory
paragraphs It is a delusion to
think that it is possible to reduce
biological data to simple numerical
levels. At best, these can only be
produced for limited situations and
-------
The Interpretation of Biological Data with Reference to Water Quality
TABLE 5
1 2
Mfi
SW.l
cecKBism
1•«
aid
rlftbt
la ft
¦id rljtbt
la ft
aid rlKht
iMtkaxdu
Daadroacaa
p
c
7
SpdagLllft frafUla
Ill47l
Ifcdteatlflid 89am
c
c
P
A
A
Ccrtylophcr* lacoatarl*
f
p
A
A
Dajcnla tlKrlta
?
e
k
k
6
)b
Ureat« 11* gracilis
PtludlcaLl* «rtlcul»u
Pred*rle*Ua cuius*
c
c
c
C
c
c
p
T
P
P
T
r
P
P
Prlatioa
B&la calami*
fwimla
1
1
1
7
ll&ldeotlfltd (Mtb
1
Unidentified Boatla
1
Qtwbant* puaetlpamlj
ZftitHUi—uuj a
Crlcotopua blclnetoi
Oaldeotlrtrt TndlpadlAl
BamUchla «p A
Troll pa¦ nejYoaua
TvsOlpM aodaataa
tol/jwdliia a? B
CclocmrLr*
X
59
*T
5
Ik
5
2
16
2
9
7
5
1
11T
29
fc
12
66
1
2
15
7
W
1
1
1
26
1
8
2
k
1
3
t
101
TrjcarTtbetes
Btenonaaa
1
1
1
1
1
Agngrlra
Athrlp*odaa
flira
HjdropeTti* orrti
Qwuantoparcba
PaycboErlldee G*soa A
1
51 22
2
n
2
11
1*1
It
10
5
17
*3
92
1
19
2
11
1
131
3 u
30 13
191 21
UlhdJla TUTBCOSS
TcttIiiU ehleetll
Qos&mIa ap
Qtmilrala v&emlMt*
Carblcul* flaalm
i
1
* 1 5
1 "
33
1
Z
1 1
7'5 19
1
TT ' 7
TOTAL
15? 90 S6 . rit
3B1
Xkl 175 e»TI
298 871
1.151
P - Twv C - comao A - aboDdant
Benthos from Pickwick Tailwater (35)
even then they need verbal exposition,
at worst, they give a spurious im-
pression of having absolute validity
8 Comparison of stations
My final point in this section con-
cerns comparisons It is claimed
that the German system, in effect,
measures an absolute state, a
definite level of water quality We
have seen that this is not a tenable
claim In the other systems, by
and large, the need to establish
local control stations at which to
measure the normal or "natural"
biotic conditions is accepted, and
then other areas are compared with
this supposed norm. This is, of
course not always possible as there
may remain no unaffected area, or
no unaffected area that is, with
respect to such factors as current.
nature of substratum, etc ,
sufficiently similar to act as a
base-line for data Nevertheless,
basically, these systems can be
used to compare stations and thus
to assess changes in water quality
In doing this, they can all be used
more or less successfully, but I
maintain that a table is just as use-
ful as an elaborate analysis, and
I believe that the table should be
included with whatever is done
For a particular situation, however,
it is often possiole to distill the data
into a single figure as a measure of
similarity between stations
9 Coefficients of similarity
Burlington (1962) and Dean and Bur-
lington (1963) have recently proposed
an entirely objective means of doing
this, which involves simple arith-
metical manipulation In his system,
a "prominence value" is calculated
for each species at each station
This is a product of its density and
some function of its frequency in
samples, but the details of this
calculation can be altered to suit
any particular situation Then a
coefficient of similarity between
each pair of stations can be calcu-
lated by dividing twice the sum of
the lower prominence values of taxa
that the two stations have in common
by the sum of all the prominence
values of both stations. Identical
stations will then have a coefficient
of similarity of 1-00, this coefficient
will be lower the more the stations
differ from one another. This is an
easy way to compare stations in an
entirely unbiased way and as such
may satisfy the need for numerical
exposition, however, it tells one
nothing about why the localities are
different and like all the other more
or less numerical methods of pre-
senting data has no absolute value
Moreover, it still leaves unanswered
the fundamental question of how
different is "different9 "
-------
The Interpretation of Biological Data with Reference to Water Quality
TABLE 6
^Clean
Types of
Organisms
BIOTIC INDEX1
VTotal Number of Groups
Present
Present
Variety Present
0-1
2-5
6-10
11-15
16+
0)
*u «d
Plecoptera
nymph
More than one species
—
Bio
7
Inde>
8
9
10
present
One species only
~
6
7
8
9
Ephemeroptera
nymph
present
More than one species
—
6
7
8
9J
T3 cx
c «d o
t-i 01 <0
One species only
—
5
6
7
8
V Q 0)
03 M
3 O O
U C
Trichoptera
larvae
More than one species*
--
5
6
7
8
H ^
a o c
•H CO
present
One species only *
4
4
5
6
7
JJ 0)
rH U
Gammaridae
All above species absent
3
4
5
6
7
B
-------
The Interpretation of Biological Data with Reference to Water Quality
IV THE PROBLEMS OF SAMPLING
The systems outlined above are all based on
the assumption that it is possible to sample
an aquatic habitat with some degree of
accuracy, this is a dubious assumption,
however, when applied to biological data.
From what has been said about the com- B
plexity of biological reactions to the various
factors in the environment, and from the
obvious fact that rivers especially are a
mosaic of microhabitats, it is clear that to
achieve numerical accuracy or even some
limits of confidence considerable numbers
of samples need to be taken. Indeed, even
in so apparently unvaried a habitat as a
single riffle, Needham and Usinger (1956)
showed that a very large number of samples
would be necessary to give significant
numerical data.
A Representative Sampling
There is a limit to the number of samples
that can reasonably be taken and, any-
way, it is desirable to sample many
different types of habitat so as to get as
broad as possible an estimate of the
biota This is the more recent approach
of most of the workers in Central
Europe, who have been content to cite
abundances on a simple relative but
arbitrary scale and to convert this to
figures on some sort of logarithmic
scale for use in calculations. An C
alternative is to express the catch in
terms of percentage composition, but
this has the disadvantage that micro-
and macro-organisms cannot be expressed
on the same scale as they are obtained
by different collecting techniques. Also,
of course, implicit in this approach is
the assumption that the sampling is
reasonably representative. Here again
we run into the need for knowledge and
expertise. In collection as well as in
interpretation, the expert is essential.
Biological sampling, unlike the simple,
or fairly simple, filling of bottles for
chemical analysis or the monitoring of
measuring equipment, is a highly
skilled job and not one to be handed over
to a couple of vacationing undergraduates
who are sent out with a Surber sampler
and told to get on with it. This point
has also been made by other biologists,
e.g., Patrick (1961) who stresses the
need for skilled and thorough collecting
even for the determination of a species
list.
Non-Taxonomic Techniques
Alternatively we can use the less
expert man when concentrating on only
part of the habitat, using, say,
microscopical slides suspended in the
water to study algal growth. This
method was extensively used by
Butcher (1946), and Patrick et al.(1954)
who studied diatoms in this way.
This gives only a partial biological
picture, but is useful as a means of
monitoring a stretch of water where it
is possible that changes might occur.
It is a useful short-hand method, and
as such is perhaps comparable to
studying the oxygen absorbed from
potassium permanganate instead of
carrying out all the usual chemical
analyses on water. A short method of
this kind may serve very well most of
the time, but, for instance, would not
be likely to detect an insecticide in
concentrations that could entirely
eliminate arthropods and hence fishes
by starvation.
Monitoring
It is possible to work out biological
monitoring systems for any specific
purpose. The simplest of these is the
cage of fish, which, like a single type
of chemical analysis, can be expected
to monitor only one thing -- the ability
of fish to live in the water -- with no
information on whether they can breed
or whether there is anything for them
to eat. Beak et al. (1959) describe a
neat way in which the common con-
stituents of the bottom fauna of Lake
Ontario can be used to monitor the
effluents from an industrial site.
Obviously there is much room for such
ingenuity m devising biological systems
for particular conditions, but this is
perhaps outside the scope of this meeting
-------
The Interpretation of Biological Data with Reference to Water Quality
V CONCLUSIONS B
It may appear from the previous sections
that my attitude to this problem is entirely
obstructionist. This is far from being so.
Water quality is as much biological phenom-
enon as it is a chemical or physical one,
often what we want to know about water is
almost exclusively biological -- will it smell
nasty, is it fit to drink, can one bathe in it,
etc' I suggest, therefore, that it is desirable
to organize water monitoring programs that
will tell one what one wants to know. There
is no point in measuring everything biolog-
ical, just as there is no point in performing
every possible chemical analysis, what is
measured should be related to local conditions.
It would be a waste of time to measure
oxygen content in a clean mountain stream,
we know it to be high, and it becomes worth
measuring only if we suspect that it may C
have been lowered by pollution. Similarly,
there is little point in studying the plankton
in such a stream, we know it only reflects
the benthic flora. In a lake or in a slow
river, on the other hand, if our interest in
the water lies in its potability, records of
the plankton are of considerable importance
as changes in plankton are, in fact, changes
in the usability of the water.
A Periphyton and Benthos Studies
For long-term studies, especially for
the recording of trends or changes
induced by pollution, altered drainage,
agricultural poisons, and other havoc
wrought by man, one can expect in-
formative results from two principal
techniques. First, we can study
microscopic plant and animal growth
with glass slides placed in the water for
fixed periods, second, we can obtain
random samples of the benthic fauna.
The algae and associated microfauna
tell one a good deal about the nutrient
condition of the water and the changes
that occur in it, and the larger benthic
fauna reveal changes in the trophic
status, siltation due to soil erosion,
effects of insecticides and other poisons,
etc.
Varying Levels of Complexity
The study of growths on glass slides is
reasonably skilled work, but can easily
be taught to technicians, like chemical
monitoring, such study needs to be
done fairly often. Sampling the benthos
is more difficult and, as explained
above, needs expert handling, unlike
most other monitoring programs,
however, it need be done only in-
frequently, say, once or twice a year.
Inevitably sampling methods will vary
with type of habitat, in each case, the
question will arise as to whether it is
worth looking at the fish also. It is
here that the biologist must exercise
judgment in devising and carrying out
the sampling program.
Data Interpretation
Judgment is also needed in the inter-
pretation of the data. It is for this
reason I maintain that it should all be
tabulated so that it remains available
for reassessment or comparison with
later surveys. If need be, some sort
of numerical format can be prepared
from the data for ad hoc uses, but it
should never become a substitute for
tabulations. Only m this way can we
go on building up our knowledge.
Perhaps some day we shall be able to
pass all this information into a com-
puter, which will then be able to
exercise better judgment than the
biologist. I hope this will happen, as
computers are better able to remember
and to cope with complexity than men.
It will not, however, pension off the
biologist. He will still be needed to
collect and identify the samples.
I cannot imagine any computer wading
about on rocky riffles nor persuading
outboard motors and mechanical grabs
to operate from the unstable confines
of small boats. We shall still need
flesh and blood biologists long after the
advent of the hardware water chemist,
even though, with reference to my
earlier analogy, a Tokyo University
-------
The Interpretation of Biological Data with Reference to Water Quality
computer recently outpointed 10 veteran
medicals in diagnosing brain tumors and
heart disease. It should be pointed out,
however, that the computer still had to be
fed with information, so we are still
a long way from the hardware general
practitioner. I believe though that he is
likely to evolve before the hardware
biologist; after all, he studies only one
animal.
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Kaliabwasser auf die Fauna der
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3.401-26. 1954.
2 Allanson, B. R. Investigations into the
ecology of polluted inland waters in
the Transvaal. Part I Hydrobiologia
18:1-94. 1961.
3 Bartsch, A. F. and Ingram, W. M.
Biological Analysis of "Water Pollution
in North America. Verh. Internat.
Verein. Limnol. 16:788-800. 1968.
4 Beak, TW.de Courval, C. and
Cooke, N. E. Pollution monitoring
and prevention by use of bivariate
control charts. Sew. Industr.
Wastes 31-1383-94. 1959.
5 Beck, W M. , Jr. The Use and Abuse of
Indicator Organisms. Transactions
of a Seminar on Biological Problems
m Water Pollution. Cincinnati. 1957.
6 Burlington, R. F. Quantitative Biological
Assessment of Pollution. J. Wat
Poll. Contr. Fed. 34-179-83. 1962.
7 Butcher, R. W The Biological Detection
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2:92-7. 1946.
8 Cairns, John, Jr. et al. A Preliminary
Report on Rapid Biological Information
Systems for Water Pollution Control.
JWPCF. 42(5):685-703. 1970.
9 Caspers, H. and Schulz, H. Studien
zur Wertung der Saprobiensysteme.
Int. Rev. ges. Hydrobiol. 45:535-65.
1960.
10 Dean, J. M. and Burlington, R. F.
A Quantitative Evaluation of Pollution
Effects on Stream Communities.
Hydrobiologia 21:193-9. 1963.
11 Ferdjingstad, E. Taxonomy and
Saprobic Valency of Benthic Phyto-
microorganisms. Inter. Revue der
Ges. Hydrobiol. 50(4):475-604. 1965.
12 Ferdjingstad, E. Pollution of Streams
Estimated by Benthal Phytomicro-
organisms. I. A System Based on
Communities of Organisms and
Ecological Factors. Int. Revue ges.
Hydrobiol. 49:63-131.
13 Gaufin, A. R The Effects of Pollution
on a Midwestern Stream. Ohio J.
Sci. 58:197-208. 1958.
14 Gaufin, A. R and Tarzwell, C. M.
Aquatic Invertebrates as Indicators
of Stream Pollution. Pub. Hlth.
Rep. 67-57-64. 1952.
15 Hairston, N. G. Species Abundance and
Community Organization. Ecology
40:404-15. 1959.
16 Harrison, A. D The Effects of Sulphuric
Acid Pollution on the Biology of
Streams in the Transvaal, South
Africa. Verh. Int. Ver. Limnol.
13:603-10. 1958.
17 Harrison, A. D The role of River Fauna
in the Assessment of Pollution.
Cons. Sci. Afr Sud Sahara Pub.
64:199-212. 1960.
18 Hirsch, A. Biological Evaluation of
Organic Pollution of New Zealand
Streams. N.Z. J Sci. 1-500-53.
1958.
19 Hynes, H. B. N. The Biology of
Polluted Waters. Liverpool. 1960.
20 Hynes, H.B.N. The Effect of Sheep-
dip Containing the Insecticide BHC
on the Fauna of a Small Stream.
Ann. Trop. Med. Parasit.
55:192-6. 1961.
21 Hynes, H.B.N, and Roberts, F.W.
The Biological Effects of Detergents
in the River Lee, Hertfordshire.
Ann. Appl. Biol. 50:779-90. 1962.
22 Hynes, H.B.N, and Williams, T. R
The Effect of DDT on the Fauna of
a Central African Stream. Ann. Trop.
Med. Parasit. 56:78-91 1962.
23 lilies, J. Die Lebensgemeinschaft des
Bergbaches. Wittenberg-Lutherstadt.
1961a.
-------
The Interpretation of Biological Data with Reference to Water Quality
36 Patrick, R., Hohn, M. H. and Wallace,
J H. A New Method for Determining
the Pattern of the Diatom Flora.
Not. Nat Phila. Acad. Sci. 259
12 pp. 1954.
24 lilies, J. Versuch einer allgemeiner
biozonotischen Gliederung der
Fliessgewasser. Int. Rev. ges
Hydrobiol. 46:205-13. 1961b.
25 Ingram, W. M., Mackenthun, K. M. , and
Bartsch, A. F Biological Field
Investigative Data for Water Pollution
Surveys. USDI, FWPCA Pub. WP-13,
139 pages. 1966.
28 Kaiser, E. W. Biolgiske, biokemiske,
bacteriologiske samt hydrometriske
undersogelser af Poleaen 1946 og
1947. Dansk. Ingenforen. Skr.
3:15-33. 1951.
27 Keup, Lowell E. , Ingram, W M. , and
Mackenthun, K. M. Biology of Water
Pollution. USDI. FWPCA CWA-3,
290 pages. 1967.
28 Kolkwitz, R Oekologie der Saprobien.
Uber die Beziehungen der Wasser-
organismen zur Ummelt. Schr.
Reihe ver Wasserhyg. 4:64 pp. 1950.
29 Liebmann, H. Handbuch der Fnschwasser
und Abwasserbiologie. Munich. 1951.
30 Maciel, Norma C Levantamento
hipotetico de um rio com rede
Surber. Inst, de Engenharia Sanitaria,
Rio de Janeiro, Brazil. Pub. No. 58,
96 pages. 1969. (Zones of pollution
in a Brazilian river.)
31 Mackenthun, K. M. The Practice of
Water Pollution Biology. USDI.
FWPCA. 281 pp. 1969.
32 Needham, P. R and Usmger, R. L.
Variability in the Macrofauna of a
Single Riffle in Prosser Creek,
California, as indicated by the Surber
Sampler. Hilgardia 24:383-409. 1956.
33 Olson, Theodore A., and Burgess, F. J.
Pollution and Marine Ecology. Inter-
science Publishers. 364 pages. 1967.
34 Patrick, R. A Proposed Biological
Measure of Stream Conditions, based
on a Survey of the Conestoga Basin,
Lancaster County, Pennsylvania.
Proc. Acad. Nat. Sci. Phila.
101:277-341. 1949.
35 Patrick, R. A Study of the Numbers and
Kinds of Species found in Rivers in
Eastern United States. Proc. Acad.
Nat Sci. Phila. 113:215-58. 1961.
37 Patrick, Ruth. Benthic Stream Com-
munities. Amer. Sci. 58:546-549.
1970.
38 Richardson, R. E. The Bottom Fauna of
the Middle Illinois River, 1913-1925,
Its Distribution, Abundance, Valuation
and Index Value in the Study of Stream
Pollution. Bull. 111. Nat. Hist. Surv.
17-387-475. 1929.
39 Sinclair, Ralph M. , and Ingram,
William M. A New Record for the
Asiatic Clam in the United States--
The Tennessee River. Nautilus
74(3) 114-118. 1961. (A typical
benthos faunal list for a large inland
unpolluted river, with an eroding
substrate.)
40 Sladecek, Vladimir. Water Quality
System. Verh. Internat. Verein.
Limnol. 16-809-816. 1966.
41 Sladecek, V Zur biologischen
Gliederung der hoheren Saprobi-
tatsstufen. Arch. Hydrobiol.
58:103-21. 1961.
42 Sladecek, Vladimir. The Ecological and
Physiological Trends in the Sapro-
biology. Hydrobiol. 30:513-526.
1967.
43 Tumplmg, W. V Probleme, Methoden
und Ergenbmsse biologischer
Guteuntersuchungen an Vorflutern,
dargestellt am Beispiel der Werra.
45:513-34. 1960.
44 Whipple, G. C , Fair, G M. and
Whipple, M. C The Microscopy of
Drinking Water. New York. 1947.
45 Woodiwiss, F. S The Biological System
of Stream Classification used by the
Trent River Board Chem. and Ind. ,
pp. 443-447. March 1964.
46 Wurtz, C. B and Dolan, T. A Biological
Method Used in the Evaluation of Effects
of Thermal Discharge in the Schuylkill
River. Proc. Ind. Waste Conf. Purdue.
461-72. 1960.
-------
The Interpretation of Biological Data with Reference to Water Quality
47 Zimmerman, P. Der Einfluss auf die
Z usammensetzung der Lebensgemein-
schaften in Experiment. Schweiz. Z.
Hydrol. 24:408-11. 1962.
48 Hynes, H. B. N. The Ecology of Flowing
Waters in Relation to Management.
JWPCF. 42(3):418-424. 1970.
49 Hynes, H. B. N. The Ecology of Running
Waters. Univ. of Toronto Press 555 pp.
1970.
50 Scott, Ralph D. The Macro-invertebrate
Biotic Index - A Water Quality Measure-
ment and Natural Continuous Stream
Monitor for the Miami River Basin.
17 pp. The Miami Conservancy District,
Dayton, OH 45402. 1969.
51 Cooke, Norman E. Stream Surveys
Pinpoint Pollution. Industrial Water
Engineering, p. 31-33, Sept. 1970.
This outline was prepared by Dr. H.B.N.
Hynes, Chairman, Department of Biology,
University of Waterloo, Ontario, Canada.
Reprinted from Symposium Environmental
Measurements Valid Data and Logical
Interpretation, July 1964, PHS Publication
No. 999-AP-15, pp. 289-298.
Figures, tables, additional references, and
headings are editorial changes by Ralph
Sinclair, Aquatic Biologist, National Training
Center, Water Programs Operations, EPA,
Cincinnati, OH 45268.
-------
SECTION F
WATER QUALITY AND AQUATIC LIFE
Multiple usage of water is associated with a variety of concepts of acceptable
quality. Water of quality suitable for one kind of vise is not necessarily
acceptable for another kind of use.
Included in the following outlines are sections from Water Quality Criteria
(FWPCA 1968), and Water Research Needs (FWPCA 1969).
Contents of Section F
Section in - Fish, Other Aquatic Life, and Wildlife
Research Needs - Fish, Other Aquatic Life, and Wildlife
Outline No.
29
-------
fish, other aquatic life,
and wildlife
(Extracted fronts Report of the Committee on
WATER QUALITY CRITERIA
FWPCA, USDI, April 1, 1968)
WP.POL.12.7. 69
-------
letter
from the chairman
THE MEMBERSHIP of the National Techni-
cal Advisory Subcommittee on Water Quality
Criteria for Fish, Other Aquatic Life, and Wild-
life represents training and experience in several
phases of freshwater, marine and wildlife ecology,
physiology, and toxicology. The task of this Sub-
committee is to describe, insofar as possible, un-
der present knowledge: (1) the environmental
requirements of aquatic life and wildlife, (2) the
environmental concentrations of potential toxi-
cants that are not harmful under long-term ex-
posure, and (3) to suggest indirect methods for
determining safe concentrations through bioassays
and application factors. Because present knowl-
edge of environmental requirements is incomplete
and information on safe concentrations of toxi-
cants is nonexistent for most organisms, the recom-
mendations for water quality criteria of necessity
are incomplete, tentative, and subject to change as
additional information becomes available. In the
determination of these criteria, the Subcommittee
has utilized the broad knowledge, the many years
of experience, and the understanding and com-
monsense of the Subcommittee members.
In order to expedite this task, the Subcommittee
was divided into three groups: one for freshwater
organisms; one for marine and estuarine orga-
nisms; and a third for wildlife.
Six task forces were set up in each of the first
two groups. Each of these task forces was assigned
certain environmental factors to review present
knowledge and determine environmental condi-
tions essential for the survival, growth, reproduc-
tion, general well-being, and production of a
desired crop of aquatic organisms. Members as-
signed to each task force were experts on that
particular subject or had wide experience with the
factors or materials in question. They were se-
lected with this in mind so that the whole subject
could be covered most effectively. The composite
report thus prepared was reviewed by the full
Subcommittee and approved on October 31 and
November 1, 1967, in Washington, D.C.
29-2
Clarence M. Tarzwell,
-------
DURING the course of geologic time, orga-
nisms which were able to adapt so they were
better fitted to live under existing environmental
conditions were the ones which survived and now
form the biota. Geologic change is a slow process
and biota developed which were adapted not only
to the physical and chemical but also to the bio-
logical factors of the environment. The environ-
mental factors to which organisms adapted through
the evolutionary process are now their environ-
mental requirements. Therefore, any relatively
rapid change in these conditions can be detri-
mental or even disastrous. Because the biota is the
result of long evolutionary processes during which
delicate balances were established, a change in
conditions or in a portion of the biota can have
far reaching effects.
Man has now attained the ability to alter
drastically his environment and that of other
organisms Many of his activities already have im-
paired seriously his own environment and that of
other living things. Water pollution engineering
works and other changes that modify the aquatic
environment rank high in capping detrimental
effects.
Water pollutants may be harmful through alter-
ations in natural environmental conditions (such
as temperature, dissolved oxygen, pH, carbonates,
etc.), through physiological and other changes due
to the addition of toxicants, or through both. Thus,
in determining the effects of pollutants we must
consider environmental, physiological, and ac-
cumulative effects.
Substances in suspension and solution, whether
solid, liquid, or gas, largely determine the quality
of the water. Aquatic organisms are affected not
only directly by these materials, but also indirectly
through their effects on other forms of aquatic life
which comprise their food, competitors, and pred-
ators. Hence, the determination of water quality
requirements for aquatic life is a very involved
task. The problem is further complicated by the
fact that different species and different develop-
mental or life stages of the same or different
species may differ widely in their sensitivity or
tolerance to different materials, to ranges in en-
vironmental conditions, and to the cumulative
synergistic and antagonistic effects of toxicants
In determining water quality requirements for
aquatic life and wildlife, it is essential to recognize
that there are not only acute and chronic toxic
levels but also tolerable, favorable, and essential
levels of dissolved materials. Lethal, tolerable, and
favorable levels and conditions may be ascertained
by: (1) determining the environmental factors and
concentrations of materials which are favorable in
-------
natural waters; (2) determining by laboratory
studies the relative sensitivity of organisms to
various environmental factors, and ranges which
are tolerable and favorable; (3) determining by
means of different bioassay studies the behavioral,
physiological, and other responses of organisms to
potential toxicants and concentrations of these
materials which are not harmful under continuous
exposure; and (4) testing laboratory findings in
the field to determine their adequacy for the pro-
tection of aquatic and wildlife resources.
In approaching this problem of protecting our
aquatic and wildlife resources, it must also be
realized that: (1) certain natural complexes of
dissolved materials to which aquatic organisms
have become adapted are favorable whereas other
concentrations or compositions may not be; (2)
unnatural materials added by man can be unfa-
vorable; (3) altering the amounts of substances
normally found in the environment can be harm-
ful; (4) toxicity is a quantitative term—any mate-
rial becomes toxic when its concentration exceeds
certain levels. It is essential, also, to realize that
requirements must be maintained throughout pe-
riods of low water, maximum discharge, maximum
temperature, minimum DO, variations in pH,
turbidity, salinity, etc Further, it should be under-
stood that: (1) unfavorable conditions which may
be resisted for long periods by adults may be en-
tirely unfavorable for the survival of the species;
(2) conditions need to be unfavorable for only a
few hours to eliminate a population or group of
species; and (3) levels of environmental factors
and concentrations of toxicants that appear to
cause no harm during a few hours of exposure
may be intolerable for extended periods or for
recurring short-term exposures.
In defining water quality requirements for
aquatic life and wildlife, it is necessary to define
the extreme upper and lower limits of the various
environmental factors as well as the optimum
values. These extremes are outer limits and con-
stitute the minimum objectives to be obtained in
the improvement of waters for aquatic life. It is
not the intention of the Subcommittee that such
levels are to be considered as satisfactory. Fur-
ther, it is stressed that waters of higher quality
should not be degraded towards approximation of
the extremes. For example, the dissolved oxygen
content of water should be near saturation for best
production. The lower limits for oxygen indicated
in the report, therefore, represents the objective to
be obtained in the improvement of water, and not
the level to which good waters may be lowered.
It is essential that the various recommendations be
considered in context with the body of the report,
taking due consideration of the variability of local
conditions and native biota.
Within the United States there are great varia-
tions in environmental conditions and in the flora
and fauna. The environmental requirements of the
biota are different not only for different regions but
for different portions of the same region Overlying
these differences are seasQnal changes and daily
variations that have become essential factors in the
environment. Ideally, therefore, water quality cri-
teria for aquatic life and wildlife should take into
consideration local variations in requirements, sea-
sonal changes, and daily variations They should
be national in scope They should be applicable to
streams of various size and character, to all types
of lakes, to reservoirs, estuaries, and coastal
waters.
It is obvious that more research is needed on the
character, conditions, and interrelations in fresh
water, marine, and estuarine ecosystems which are
subjected to degradation or alteration as well as on
the physiological requirements and tolerances of
the various species involved in these different eco-
systems This need must be satisfied for the estab-
lishing of sound criteria to maintain and preserve
aquatic resources and to permit the most economi-
cal and productive use of these resources by man.
Further, water quality requirements must be
expressed so as to allow for environmental modifi-
cations where such modifications are justifiable
and deemed to be in the public interest
All these factors have been considered m de-
veloping the following recommended water quality
requirements for aquatic life.
It is the purpose of this document to define the
water quality requirements which must be met to
insure a favorable environment for fish, other
aquatic life, and wildlife. This report will do this
by identifying those aspects of water quality that
are most important in the light of current knowl-
edge and quantifying them where possible Where
quantification is not yet possible, narrative guide-
lines will be offered There is no doubt that the
water quality requirements contained herein must
be reviewed periodically and updated in the light
of additional and improved scientific data. The
recommendations given in this report are consid-
ered to be satisfactory for aquatic life. In all
instances where natural conditions fall outside the
recommended ranges, this environment may be
marginal and should not be changed in such a way
as to make it more unfavorable
-------
zones of passage
ANY BARRIER to migration and the free
movement of the aquatic biota can be
harmful in a number of ways. Such barriers block
the spawning migration of anadromous and cata-
dromus species Many resident species make local
migrations for spawning and other purposes and
any barrier can be detrimental to their continued
existence. The natural tidal movement in estuaries
and downstream movement of planktonic orga-
nisms and of aquatic invertebrates in flowing fresh
waters are important factors in the re-population
of areas and the general economy of the water.
Any chemical or thermal barrier destroys this
valuable source of food and creates unfavorable
conditions below or above it
It is essential that adequate passageways be pro-
vided at all times for the movement or drift of the
biota. Water quality criteria favorable to the
aquatic community must be maintained at all
times in these passageways. It is recognized, how-
ever, that certain areas of mixing are unavoidable.
These create harmfully polluted areas and for this
reason it is essential that they be limited in width
and length and be provided only for mixing. The
passage zone must provide favorable conditions
and must be in a continuous stretch bordered by
the same bank for a considerable distance to allow
safe and adequate passage up and down the
stream, reservoir, lake, or estuary for free-floating
and drift organisms.
The width of the zone and the volume of flow
in it will depend on the character and size of the
stream or estuary. Area, depth, and volume of flow
must be sufficient to provide a usable and desirable
passageway for fish and other aquatic organisms.
Further, the cross-sectional area and volume of
flow in the passageway will largely determine the
percentage of survival of drift organisms. There-
fore, the passageway should contain preferably
75 percent of the cross-sectional area and/or
volume of flow of the stream or estuary. It is
evident that where there are several mixing areas
close together they should all be on the same side
so the passageway is continuous. Concentrations
of waste materials in passageways should meet the
requirements for the water.
The shape and size of mixing areas will vary
with the location, size, character, and use of the
receiving water and should be established by
proper administrative authority. From the stand-
point of the welfare of the aquatic life resource,
however, such areas should be as small as possible
and be provided for mixing only Mixing should be
accomplished as quickly as possible through the
use of devices which insure that the waste is mixed
with the allocated dilution water in the smallest
possible area. At the border of this area, the water
quality must meet the water quality requirements
for that area. If, upon complete mixing with the
available dilution water these requirements are not
met, the waste must be pretreated so they will be
met. For the protection of aquatic life resources,
mixing areas must not be used for, or considered
as, a substitute for waste treatment, or as an exten-
sion of, or substitute for, a waste treatment facility.
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RECOMMENDATIONS given below are con-
sidered to be satisfactory for aquatic life. In
all instances where natural conditions fall outside
the recommended ranges, these conditions may
be marginal and should not be changed in such a
way as to make them more unfavorable
summani
and key
criteria
Freshwater organisms
Dissolved Materials
(1) Dissolved materials that are relatively in-
nocuous; i e , their harmful effect is due to osmotic
effects at high concentrations, should not be in-
creased by more than one-third of the concentra-
tion that is characteristic of the natural condition
of the subject water. In no instance should the
concentration of total dissolved materials exceed
50 milliosmoles (the equivalent of 1500 mg/1
NaCl)
(2) Dissolved materials that are harmful in
relatively low concentrations are discussed in the
section "Toxicity."
pH, Alkalinity, Acidity
(1) No highly dissociated materials should be
added in quantities sufficient to lower the pH be-
low 6.0 or to raise the pH above 9.0.
(2) To protect the carbonate system and thus
the productivity of the water, acid should not be
added in sufficient quantity to lower the total al-
kalinity to less than 20 mg/1.
(3) The addition of weakly dissociated acids
and alkalies should be regulated in terms of their
own toxicities as established by bioassay pro-
cedures.
Temperature
Warm Water Biota: To maintain a well-
rounded population of warm-water fishes, the fol-
lowing restrictions on temperature extremes and
temperature increases are recommended:
(1) During any month of the year heat should
not be added to a stream in excess of the amount
that will raise the temperature of the water (at the
expected minimum daily flow for that month)
more than 5 F. In lakes, the temperature of the
epilimnion in those areas where important orga-
nisms are most likely to be adversely affected
should not be raised more than 3 F above that
which existed before the addition of heat of artifi-
cial origin The increase should be based on the
monthly average of the maximum daily tempera-
ture Unless a special study shows that a discharge
-------
of a heated effluent into the hypolimnion will be
desirable, such practice is not recommended and
water for cooling should not be pumped from the
hypolimnion to be discharged to the same body of
water.
(2) The normal daily and seasonal tempera-
ture variations that were present before the addi-
tion of heat due to other than natural causes
should be maintained.
(3) The recommended maximum temperatures
that are not to be exceeded for various species of
warm-water fish are given in table 111-1.
Cold Water Biota: Because of the large number
of trout and salmon waters which have been de-
stroyed, made marginal, or nonproductive, remain-
ing trout and salmon waters must be protected if
this resource is to be preserved.
Inland trout streams, headwaters of salmon
streams, trout and salmon lakes, and the hypolim-
nion of lakes and reservoirs containing salmonids
and other cold water forms should not be warmed
or used for cooling water. No heated effluents
should be discharged in the vicinity of spawning
areas.
For other types and reaches of cold-water
streams, reservoirs and lakes, the following re-
strictions are recommended:
(1). During any month of the year heat should
not be added to a stream in excess of the amount
that will raise the temperature of the water more
than 5 F (based on the minimum expected flow for
that month). In lakes, the temperature of the
epilimnion should not be raised more than 3 F by
the addition of heat of artificial origin
(2) The normal daily and seasonal temperature
fluctuations that existed before the addition of heat
due to other than natural causes should be main-
tained
TABLE lll-l
[Provisional maximum temperatures recommended as compati-
ble with the well-being of various soecies of fish and
their associated biota]
93 F: Growth of catfish, gar, white or yellow bass,
spotted bass, buffalo, carpsucker, threadfin shad,
and gizzard shad.
90 F: Growth of largemouth bass, drum, btuegill, and
crappie.
84 F: Growth of pike, perch, walleye, smallmouth bass,
and sauger.
80 F: Spawning and egg development of catfish,
buffalo, threadfin shad, and gizzard shad
75 F. Spawning and egg development of largemouth
bass, white and yellow bass, and spotted bass.
68 F: Growth or migration routes of salmonids and for
egg development of perch and smallmouth bass.
55 F: Spawning and egg development of salmon and
trout (other than take trout).
48 F: Spawning and egg development of lake trout,
walleye, northern pike, sauger, and Atlantic
salmon.
Note —Recommended temperatures for other species, not
listed above, may be «stablishied If Bnd when necessary in
formation becomes available
(3) The recommended maximum temperatures
that are not to be exceeded for various species of
cold-water fish are given in table III—1.
Dissolved Oxygen
The following environmental conditions are
considered essential for maintaining native popula-
tions of fish and other aquatic life.
(1) For a diversified warm-water biota, includ-
ing game fish, DO concentration should be above
5 mg/1, assuming normal seasonal and daily
variations are above this concentration. Under
extreme conditions, however, they may range be-
tween 5 and 4 mg/1 for short periods during any
24-hour period, provided that the water quality
is favorable in all other respects. In stratified
lakes, the DO requirements may not apply to the
hypolimnion. In shallow unstratified lakes, they
should apply to the entire circulation water mass.
These requirements should apply to all waters
except administratively established mixing zones.
In lakes, such zones must be restricted so as to
limit the effect on the biota. In streams, there must
be adequate and safe passageways for migrating
forms. These must be extensive Enough so that the
majority of plankton and other drifting organisms
are protected (see section on zones of passage).
(2) For the cold-water biota, it is desirable that
DO concentrations be at or near saturation. This
is especially important in spawning areas where
DO levels must not be below 7 mg/1 at any time.
For good growth and the general well-being of
trout, salmon, and their associated biota, DO con-
centrations should not be below 6 mg/1. Under
extreme conditions, they may range between 6
and 5 mg/1 for short periods provided the water
quality is favorable in all other respects and nor-
mal daily and seasonal fluctuations occur. In large
streams that have some stratification or that serve
principally as migratory routes, DO levels may
range between 4 and 5 mg/1 for periods up to
6 hours, but should never be below 4 mg/1 at any
time or place.
(3) DO levels in the hypolimnion of oligo-
trophic small inland lakes and in large lakes should
not be lowered below 6 mg/l at any time due to
the addition of oxygen-demanding waste or other
materials.
Carbon Dioxide
According to our present knowledge of the sub-
ject, it is recommended that the "free" carbon
dioxide concentration should not exceed 25 mg/1.
Oil
Oil or petrochemicals should not be added in
such quantities to the receiving waters that they
will—
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(1) produce a visible color film on the surface;
(2) impart an oily odor to the water or an oily
or other noxious taste to fish and edible in-
vertebrates;
(3) coat the banks and bottoms of the water
course or taint any of the associated biota;
(4) become effective toxicants according to the
criteria recommended in the "Toxicity"
section.
Turbidity
(1) Turbidity in the receiving waters due to the
discharge of wastes should not exceed 50 Jackson
units in warm-water streams or 10 Jackson units
in cold-water streams.
(2) There should be no discharge to warm-
water lakes which would cause turbidities exceed-
ing 25 Jackson units The turbidity of cold-water
or oligotrophic lakes should not exceed 10 units.
Settleable Materials
Since it is known that even minor deposits of
settleable materials inhibit the growth of normal
stream and lake flora, no such materials should be
added to these waters in quantities that adversely
affect the natural biota.
Color and Transparency
For effective photosynthetic production of oxy-
gen, it is required that 10 percent of the incident
light reach the bottom of any desired photosynthe-
tic zone in which adequate dissolved oxygen con-
centrations are to be maintained
Floating Materials
All floating materials of foreign origin should be
excluded from streams and lakes.
Tainting Substance
AU materials that will impart odor or taste to
fish or edible invertebrates should be excluded
from receiving waters at levels that produce
tainting.
Radionuclides
(1) No radioactive materials should be pres-
ent in natural waters as a consequence of the fail-
ure of an installation to exercise appropriate con-
trols to minimize releases.
(2) No radionuclide or mixture of radionu-
clides should be present at concentrations greater
than those specified by the USPHS Drinking Water
Standards.
(3) The concentrations of radioactive mate-
rials present in fresh, estuarine, and marine waters
should be less than those that would require re-
strictions on the use of organisms harvested from
the area to meet the Radiation Protection Guides
recommended by the Federal Radiation Council.
Plant Nutrients and Nuisance Growths
The Subcommittee wishes to stress that the con-
centrations set forth are suggested solely as guide-
lines and the maintenance of these may or may not
prevent undesirable blooms. All the factors caus-
ing nuisance plant growth and the level of each
which should not be exceeded are not known.
(1) In order to limit nuisance growths, the
addition of all organic wastes such as sewage, food
processing, cannery, and industrial wastes contain-
ing nutrients, vitamins, trace elements, and growth
stimulants should be carefully controlled. Further-
more, it should be pointed out that the addition of
sulfates or manganese oxide to a lake should be
limited if iron is present in the hypolimnion as
they may increase the quantity of available
phosphorus ^
(2) Nothing should be added that causes an in-
creased zone of anaerobic decomposition of a lake
or reservoir.
(3) The naturally occurring ratios and amounts
of nitrogen (particularly NOa and NH,) to total
phosphorus should not be radically changed by the
addition of materials As a guideline, the concen-
tration of total phosphorus should not be increased
to levels exceeding 100^tg/l in flowing streams or
50 fig/l where streams enter lakes or reservoirs.
(4) Because of our present limited knowledge
of conditions promoting nuisance growth, we must
have a biological monitoring program to determine
the effectiveness of the control measures put into
operation. A monitoring program can detect in
their early stages the development of undesirable
changes in amounts and kinds of rooted aquatics
and the condition of algal growths. With periodic
monitoring, such undesirable trends can be de-
tected and corrected by more stringent regulation
of added organics
Toxic Substances
(1) Substances of Unknown Toxicity: All efflu-
ents containing foreign materials should be con-
sidered harmful and not permissible until bioassay
tests have shown otherwise It should be the obli-
gation of the agency producing the effluent to dem-
onstrate that it is harmless in the concentrations
to be found in the receiving waters All bioassays
should be conducted strictly as recommended in
the body of this report and the appropriate appli-
-------
cation factor applied to determine the permissible
concentration of toxicant.
(2) Pesticides.
(a) Chlorinated hydrocarbons: Any addition
of chlorinated hydrocarbon insecticides is likely to
cause damage to some desired organisms and
should be avoided.
(b) Other chemical pesticides: Addition of
other kinds of chemicals used as pesticides and
herbicides can cause damage to desirable orga-
nisms and should be applied with utmost discretion
and caution. Table III—5 (p. 62) lists the 48-hour
TLm values of a number of pesticides for various
types of fresh water organisms. To provide rea-
sonably safe concentrations of these materials in
receiving waters, application factors ranging from
Vio to Vioo should be used with these values
depending on the characteristic of the pesticide in
question and used as specified in (4), below.
Concentrations thus derived may be considered
tentatively safe under the conditions specified.
(3) Other Toxic Substances.
(a) ABS: Concentration of continuous expo-
sure to ABS should not exceed y, of the 48-hour
TLm A concentration as high as 1 mg/I may be
tolerated occasionally for periods of time not ex-
ceeding 24 hours. ABS may increase the toxicity
of other materials.
(b) LAS: The concentration of LAS should
not exceed 0.2 mg/1 or % of the 48-hour TLm.
(4) Application Factors: Concentration of ma-
terials that are nonpersistant (that is, have a half-
life of less than 96 hours) or have noncumulative
effects after mixing with the receiving waters
should not exceed Vio of the 96-hour TLm value
at any time or place. The 24-hour average of the
concentration of these materials should not exceed
V&o of the TLm value after mixing. For other toxi-
cants the concentrations should not exceed %o
and y10o of the TLm value under the conditions
described above. Where specific application factors
have been determined, they will be used in all
instances.
(5) General Considerations. When two or more
toxic materials that have additive effects are pres-
ent at the same time in the receiving water, some
reduction is necessary in the permissible concen-
trations as derived from bioassays on individual
substances or wastes. The amount of reduction re-
quired is a function of both the number of toxic
materials present and their concentrations in re-
spect to the derived permissible concentration. An
appropriate means of assuring that the combined
amounts of the several substances do not exceed a
permissible concentration for the mixture is
through the use of following relationship:
(&+g • ¦
Where Ca, Cb, . . . Q are the measured concen-
trations of the several toxic materials in the water
and La, Lb, . . L„ are the respective permissible
concentration limits derived for the materials on
an individual basis. Should the sum of the several
fractions exceed one, then a local restriction on
the concentration of one or more of the substances
is necessary.
Marine and estuarine organisms
Salinity
To protect estuarine organisms, no changes in
channels, basin geometry, or freshwater influx
should be made which would cause permanent
changes in isohaline patterns of more than 10 per-
cent of the naturally occurring variation.
Currents
Currents are important for transporting nutri-
ents, larvae, and sedimentary materials for flushing
and purifying wastes, and for maintaining patterns
of scour and fill. To protect these functions, there
should be no changes in basin geometry or fresh-
water inflow that will alter current patterns in such
a way as to adversely affect existing biological and
sedimentological situations
PH
No materials that extend normal ranges of pH
at any location by more than 0.1 pH unit should
be introduced into salt water portions of tidal tribu-
taries or coastal waters. At no time should the in-
troduction of foreign materials cause the pH to be
less than 6.7 nor greater than 8.5.
Temperature
In view of the requirements for the well-being
and production of marine organisms, it is con-
cluded that the discharge of any heated waste into
any coastal or estuarine waters should be closely
managed. Monthly means of the maximum daily
temperatures recorded at the site in question and
before the addition of any heat of artificial origin
should not be raised by more than 4 F during the
fall, winter, and spring (September through May),
or by more than 1.5 F during the summer (June
through August), North of Long Island and in
the waters of the Pacific Northwest (north of
California), summer limits apply July through
September; and fall, winter, and spring limits ap-
-------
ply October through June. The rate of tempera-
ture change should not exceed 1 F per hour except
when due to natural phenomena.
Suggested temperatures are to prevail outside
of established mixing zones as discussed in the
section on zones of passage.
Dissolved Oxygen
Oxygen levels sufficient for the survival, growth,
reproduction, general well-being, and production
of a suitable crop must be maintained. The dis-
solved oxygen concentrations necessary to attain
this objective in coastal waters, estuaries, and tidal
tributaries are:
(1) Dissolved oxygen concentrations in surface
coastal waters should be greater than 5.0 mg/J
except when upwellings and other natural pheno-
mena may cause this value to be depressed.
(2) Dissolved oxygen concentration in estu-
aries and tidal tributaries should not be less than
4.0 mg/1 at any time or place except in naturally
dystrophic waters or where natural conditions
cause DO to be depressed.
Oil
No oil or petroleum products should be dis-
charged into estuarine or coastal waters in quanti-
ties that: (1) Can be detected as a visible film,
sheen, or by odor; (2) cause tainting of fish or
edible invertebrates; (3) form an oil sludge de-
posit on the shores or bottom of the receiving body
of water, (4) become effective toxicants according
to the criteria recommended in the "Toxicity"
section.
Turbidity
No effluent that may cause changes in turbidity
or color should be allowed to enter estuarine or
coastal waters unless it can be shown to have no
deleterious effects on the aquatic biota.
Settleable and Floating Substances
No materials that contain settleable solids or
substances that may precipitate out in quantities
that adversely affect the biota should be introduced
into coastal or estuarine waters It is especially
urgent that areas which serve as habitat or nursery
grounds for commercially important species be
protected from any impairment of natural
conditions.
Tainting Substances
Substances that taint or produce off-flavors in
fish and edible invertebrates should not be pres-
ent in concentrations discernible by bioassay or
organoleptic tests
Radionuclides
The recommendations made for freshwater or-
ganisms apply to marine and estuarine organisms.
Plant Nutrients and Nuisance Organisms
(1) No changes should be made in the basin
geometry, current structure, salinity, or tempera-
ture of the estuary until studies have shown that
these changes will not adversely affect the biota or
promote the increase of nuisance organisms.
(2) The artificial enrichment of the marine en-
vironment from all sources should not cause any
major quantitative or qualitative alteration in the
flora such as the production of persistant blooms
of phytoplankton (whether toxic or not), dense
growths of attached algae or higher aquatics, or
any other sort of nuisance that can be attributed
directly to nutrient excess or imbalance Because
these nutrients often are derived largely from
drainage from land, special attention should be
given to correct land management in river basins
and shores of embayments to control unavoidable
erosion.
(3) The naturally occurring atomic ratio of
NO,-N to P04-P in a body of water should be
maintained. Similarly, the ratio of inorganic phos-
phorus (orthophosphate) to total phosphorus (the
sum of inorganic phosphorus, dissolved organic
phosphorus, and particulate phosphorus) should
be maintained as it occurs naturally. Nutrient im-
balances have been shown to cause a change in the
natural diversity of desirable organisms and to
reduce productivity
Toxic Substances
(1) Substances of Unknown Toxicity: All efflu-
ents containing foreign materials should be con-
sidered harmful and not permissible until bioassay
tests have shown otherwise. It should be the obli-
gation of the agency producing the effluent to dem-
onstrate that it is harmless in the concentrations
that will be found in the receiving waters. All bio-
assays should be conducted strictly as recom-
mended in the body of this report and the appro-
priate application factor applied to determine the
permissible concentration of toxicant.
(2) Pesticides for Which Limits Have Been
Determined: The pesticides are grouped according
to their relative toxicity to shrimp Criteria are
based on the best estimates in the light of present
knowledge and it is to be expected that acceptable
levels of toxic materials may be changed as a result
of future research.
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Pesticide group A.—The following chemicals
are acutely toxic at concentrations of 5 fig/l and
less. On the assumption that y100 of this level rep-
resents a reasonable application factor, it is rec-
ommended that environmental levels of these
substances not be permitted to rise above 50 nano-
grams/1. This level is so low that these pesticides
could not be applied direcdy in or near the marine
habitat without danger of causing damage, The 48-
hour TLm is listed for each chemical in ^g/l.
Organochloride pesticides
Aldrin 0 04
BHC 2 0
Chlordane 2 0
Endrin — 0 2
Heptachlor 0 2
Lindane 0 2
DDT 0 6
Dieldnn 0 3
Endosulfan 0 2
Methoxychlor 4 0
Perthane 3 0
TDE 3 0
Toxaphene 3 0
Orgattophosphorus pesticides
Coumaphos 2 0 Naled 3 0
Dursban 3.0
Fenthion 0 03
Parathion 1 0
Ronnel 5 0
Pesticide group B.—The following types of
pesticide compounds are generally not acutely
toxic at levels of 1.0 mg/1 or less. It is recom-
mended that an application factor of Vioo be used
and in the absence of acute toxicity data that an
environmental level of not more than 10 p.g/\
be permitted An acute toxicity factor must be es-
tablished for each specific chemical in this group
to determine that it is not more toxic than related
compounds as indicated above-
Arsenicals
Botanicals
Carbamates
2,4-D compounds
2,4,5-T compounds
Phthalic acid compounds
Triazine compounds.
Substituted urea compounds.
Other Pesticides.—Acute toxicity data are avail-
able for approximately 100 technical-grade pesti-
cides in general use not listed in the above groups.
These chemicals are either not likely to reach the
marine environment or, if used as directed by the
registered label, probably would not occur at levels
toxic to marine biota It is presumed that criteria
established for these chemicals in fresh water will
protect adequately the marine habitat It should
be emphasized that no unlisted chemical should
be discharged into the estuary without preliminary
bioassay tests.
(3) Industrial and Other Toxic Wastes,
(a) Safe concentrations of metals, ammonia,
cyanide, and sulfide should be determined by the
use of appropriate application factors to 96-hour
TLm values as determined by flow-through bioas-
says using dilution water that came from the re-
ceiving body. Test organisms should be local
species or life stages of organisms of economic
and ecologic importance which are the most sensi-
tive to the waste in question. Application factors
should be ]/i o<> rnetals, y20 for ammonia,
for cyanide, and %0 for sulfide.
(b) Fluoride concentrations should not exceed
those for drinking water.
(c) Permissible levels of detergents in fresh
waters should also be applied to the marine and
estuarine waters
(d) Bacteriological criteria of estuarine waters
utilized for shellfish cultivation and harvesting
should conform with the standards as described in
the National Shellfish Sanitation Program Manual
of Operation. These standards provide that—
(1) examinations shall be conducted in accord-
ance with the American Public Health Association
recommended procedures for the examination of
sea water and shellfish,
(2) there shall be no direct discharges of un-
treated sewage,
(3) samples of water for bacteriological exami-
nation to be collected under those conditions of
time and tide which produce maximum concentra-
tion of bacteria;
(4) the coliform median MPN of the water
does not exceed 70/100 ml, and not more than
10 percent of the samples ordinarily exceed an
MPN of 230/100 ml for a five-tube decimal dilu-
tion test (or 330/100 ml, where the three-tube
decimal dilution test is used) in those portions of
the area most probably exposed to fecal contami-
nation during the most unfavorable hydrographic
and pollution conditions;
(5) the reliability of nearby waste treatment
plants shall be considered in the approval of areas
for direct harvesting.
(e) Wastes from tar, gas, coke, petrochemical,
pulp and paper manufacturing, waterfront and
boating activities, hospitals, marine laboratories
and research installation wastes are all complex
mixtures having great variability in character and
toxicity. Due to this variability, safe levels must be
determined at frequent intervals by flow-through
bioassays of the individual effluents.
For those operations having persistent toxicants,
an application factor of Vioo should be used while
for those composed largely of unstable or biode-
gradable toxicants, an application factor of V20 is
tentatively suggested.
(4) General Considerations.—When two or
more toxicants that have additive effects are pres-
ent, they must be treated as suggested earlier under
fresh water organisms.
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Wildlife
Dissolved Oxygen
In addition to the DO requirements for aquatic
organisms, the bottoms of areas used by wildfowl
must be kept aerobic to suppress botulinus
organisms.
P«
Aquatic plants of greatest value as food for
waterfowl thrive best in waters with a summer pH
range of 7.0 to 9 2.
Alkalinity
Waterfowl habitats, to be productive, should
have a bicarbonate alkalinity between 30 and 130
mg/1. Fluctuations should be less than 50 mg/1
from natural conditions.
through biological magnification. Special consid-
eration must be given to keep edible wildlife safe
for consumption by humans
Disease
Offal from poultry houses, meatpacking plants,
as well as other possible sources of disease orga-
nisms, must be excluded from areas supporting
wildlife to guard against transmission of such dis-
eases as botulism, fowl cholera, and aspergillosis.
General
Water quality suitable for fish and other aquatic
organisms will be adequate for wildlife.
Salinity
Salinity should be kept as close to natural condi-
tions as possible. Fluctuations in salinity during
any 24-hour period should be limited as follows.
v? * a a . Variation
Natural salinity permitted
0 to 3 5%c 1%,
3 5 to 13.5^o 2%c
13 5 to 35 0r/rr 4%c
Light Penetration
Optimum light requirements for aquatic wildlife
habitats should be at least 10 percent of incident
light at the surface to a 6-foot depth; the tolerable
limit should be 5 percent of the light at the surface
to the same depth.
Settleable Substances
Settleable substances destroy the usefulness of
aquatic bottoms for waterfowl. Settleable sub-
stances should be excluded from areas expected to
support waterfowl.
Oil
Oil is an especially dangerous substance to
waterfowl. Oil and petrochemicals must be ex-
cluded from both the surface and bottoms of any
area used by waterfowl.
Toxic Substances
Toxic substances should be excluded from
wildlife habitats to the degree that they affect the
health and well-being of wildlife, either directly or
-------
Dissolved materials
fresh water
organisms
Water devoid of dissolved materials is intoler-
able in nature because pure water will not support
aquatic life. Natural waters contain endless varie-
ties of dissolved materials in concentrations that
differ widely from one locality to another as well
as from time to time. Many of these dissolved ma-
terials are essential for growth, reproduction, and
the general well-being of aquatic organisms. The
chlorides, carbonates, and silicates of sodium, po-
tassium, calcium, and magnesium are generally
the most common salts present Traces of most
other essential substances are also found.
Aquatic organisms live in different concentra-
tions of dissolved substances but productivity de-
clines as the concentrations move away from the
optimum Seldom, if ever, are the dissolved sub-
stances at the optimum concentrations as we know
them The range of tolerance may be relatively
wide, but when the concentrations reach too low or
too high a level, organisms degenerate and die
Different organisms vary in their optimum require-
ments as well as in their ability to live and thrive
under variations from the optimum Some orga-
nisms are equally at home in sea water and in
fresh water. Other organisms will tolerate only one
or the other.
Any of the substances necessary to aquatic or-
ganisms has a range of concentration that is both
essential and tolerable The tolerance levels for
any one substance vary depending on the concen-
trations of other substances present. The presence
of certain substances synergizes the effects of some
materials but antagonizes the effects of others.
Under optimal concentrations, the synergistic and
antagonistic effects are in balance and relatively
high concentrations can be tolerated without ad-
verse effects
Although several measures of dissolved mate-
rials are available, no measure in itself is adequate
as an index of optimum concentration nor is any
single measure adequate to express the range of
tolerance. The biological effects depend on the
concentrations of the individual solutes, some of
which are tolerated in terms of grams per liter but
others only in nanograms per liter Some exert con-
siderable osmotic pressure, but for others the
osmotic effect is negligible. Some substances con-
tribute greatly to conductivity, while others have
little or no effect
In general, the concentrations of dissolved ma-
terials in natural fresh waters are below the opti-
mum for maximum productivity In many in-
stances, therefore, the addition of any of a large
-------
number of substances will be beneficial. In this
way, many water courses have a capacity to ab-
sorb materials to advantage. But the addition of
what may be considered beneficial substances must
be controlled so that they will not exceed favorable
limits.
The osmotic concentration of the body'fluids of
a fresh water animal is generally the maximum
concentration of dissolved material that the ani-
mal will tolerate In some animals, notably some of
the fresh water mollusks, the body fluids have an
osmotic concentration as low as 50 milliosmoles
(the equivalent of about 0 025 molar or 1,500
mg/1 sodium chloride) If the dissolved materials
are relatively innocuous, having only an osmotic
effect, it is judged that the total dissolved materials
in a water course may be increased to a certain
extent but they should not exceed 50 milliosmoles
if the fauna is to be maintained.
Many species of diatoms are very sensitive to
changes in chloride and other salt concentrations.
Some species, such as those in mountain streams
and in black water streams of the coastal plains,
can live only in waters with extremely low concen-
trations of salts. The addition of salts to such
streams will eliminate many desirable species of
diatoms and permit undesirable species to flourish
Such changes may reduce the desirable food
sources and bring about nuisance problems as
well It is believed that the total dissolved mate-
rial in a water course should not be increased by
more than one-third of that which is characteristic
of the natural conditions of such a water course
The toxicity of substances added to natural
waters often depends on the substances already
present in the receiving waters With synergism,
the toxicity increases, and with antagonism it de-
creases. Again the reaction of the toxic substances
may produce, in some cases, new products of
greater toxicity, and in others, products of lesser
toxicity
In view of the many factors that become in-
volved in the disposal of soluble materials in na-
tural waters, it is evident that no simple answer is
available. Therefore, bioassays should be used to
determine the amounts of the materials that may
be tolerated without reducing the productivity of
the watercourse in question
Recommendation: Dissolved materials are of two
types- those that are toxic at very low concentrations
and those, such as the salts of the earth metals, that
are required in certain concentrations for a productive
water and become harmful only at high concentrations
by exerting an osmotic effect If the dissolved materials
are relatively innocuous, i e , their harmful effect is an
osmotic one at high concentrations, it is judged that the
total dissolved materials of this type may be increased
to a certain extent but they should not exceed 50 mil-
liosmoles in waters where diversified animal popula-
tions are to be protected Further, to maintain local
conditions, total dissolved materials should not be in-
creased by more than one-third of the concentration
that is characteristic of the natural condition of the
water When dissolved materials are being increased,
bioassays and field studies should be used to determine
how much of the materials may be tolerated without
reducing the productivity of the desired organisms.
Acidity alkalinity, and pH
Acidity and alkalinity are reciprocal terms
Acidity is produced by substances that yield hydro-
gen ions on hydrolysis and alkalinity is produced
by substances that yield hydroxyl ions. Other defi-
nitions state that a substance is acid if it will neu-
tralize hydroxyl ions and a substance is alkaline if
it will neutralize hydrogen ions. The terms "total
acidity" and "total alkalinity" are often used to
express the buffering capacity of a solution Acidity
in natural waters is caused by carbon dioxide, min-
eral acids, weakly dissociated acids, and the salts
of strong acids and weak bases. Alkalinity is
caused by strong bases and the salts of strong
alkalies and weak acids.
An index of the hydrogen ion activity is pH.
Even though pH determinations are used as an
indication of acidity and/or alkalinity, pH is not a
measure of either As pointed out in the first sen-
tence in the previous paragraph, acidity and al-
kalinity are reciprocal terms Indeed, a water may
have both an acidity and alkalinity at the same
time. Total acidity, by definition, is the amount of
standard alkali required to bring a sample to pH
8 3. Total alkalinity, similarly, is the amount of
standard acid required to bring a sample to pH
4 5 Both are expressed in equivalents of CaC03.
Under these circumstances, there is a relation-
ship between pH, acidity, and alkalinity since, by
definition (see Standard Methods for the Exami-
nation of Water and Wastewater, 12th edition
1965), any water with a pH of 4.5 or lower has no
measurable alkalinity and a water with a pH of
8 3 or higher has no measurable acidity.
In natural waters, where the pH is in the vicinity
of 8 3, acidity is not a factor of concern. In most
productive, fresh, natural waters, the pH falls in
the range between 6.5 and 8.5 (except when in-
creased by photosynthetic activity). Some aquatic
organisms have been found to live at pH 2 and
lower and others at pH 10 and higher; however,
such organisms are relatively few. Some natural
waters with a pH of 4 support fish and other or-
ganisms. In these cases the acidity is due primarily
to carbon dioxide and humic acids and the water
-------
has little buffering capacity (low total alkalinity).
Other natural waters with a pH of 9.5 also support
fish, but in such situations the waters are not
regarded as highly productive.
Acids that dissociate to a high degree do not
appear to be toxic at pH values above 6.0. They
are toxic if added in sufficient quantities to reduce
the pH to less than 6.0 Acids that dissociate to a
low degree are often toxic at pH values consider-
ably above 6.0. In the latter condition, toxicity is
due either to the anion or to the compound itself;
e.g, hydrogen cyanide (HCN), hydrogen sulfide
(H2S), and hypochlorous (HCIO) and tannic
acids.
Alkalies that dissociate to a high degree do not
appear to be toxic at pH values below 9.0 Alka-
line compounds that dissociate to a low degree are
often toxic at pH values less than 9.0 and their
toxicity is due either to the cation or to the undis-
sociated molecule. Ammonium hydroxide is an
example. Temporarily high pH levels often are
produced in highly productive waters through pho-
tosynthetic activity of the aquatic plants by con-
verting the carbonate to the hydroxide, which re-
sults in an increased pH. Because these high pH
levels prevail for only a few hours, they do not
produce the harmful effects of continuous high
levels due to the presence of strong alkalies.
Addition of either acids or alkalies to waters
may be harmful not only in producing adverse acid
or alkaline conditions, but also by increasing the
toxicity of various components in the waters. The
addition of strong acids may cause the formation
of carbonic acid (free C02) in quantities that are
adverse to the well-being of the organisms present.
A reduction of about 1 5 pH units can cause a
thousand-fold increase in the acute toxicity of a
metallo-cyanide complex. The addition of strong
alkalies may cause the formation of undissociated
NH
-------
Temperature
The relationships of temperature and aquatic
life have been well studied Extensive bibliogra-
phies and detailed surveys of the subject have been
published by the American Society of Civil Engi-
neers (1967), Brett (1960), Mihursky and Ken-
nedy (1967), Raney (1966), U.S Department of
Interior, Federal Water Pollution Control Admin-
istration (1967), and Wurtz and Renn (1965)
The temperatures of the surface waters of the
United States vary from 32 to over 100F as a
function of latitude, altitude, season, time of day,
duration of flow, depth, and many other variables
The agents that may affect the natural temperature
are so numerous that it seems unlikely that two
bodies of water, even in the same latitude, would
have exactly the same thermal characteristics The
fish and other aquatic life occurring naturally in
each body of water are species or varieties that are
competing there with various degrees of success
depending on the temperature and various other
conditions existing in that habitat This adaptation
extends not only to temperature and the range over
which it can vary, but also to such factors as day
length and the other sDecies of animals and plants
in the same habitat. The interrelationships of spe-
cies, day length, and water temperature are so inti-
mate that even a small change in temperature may
have far-reaching effects An insect nymph in an
artificially warmed stream, for example, might
emerge for its mating flight too early in the spring
and be immobilized by the air temperature Simi-
larly, a fish might hatch too early in the spring to
find an adequate amount of its natural food orga-
nisms because the food chain depends ultimately
on plants whose abundance in turn, is a function of
day length and temperature The inhabitants of a
water body that seldom become^ warmer than 70 F
are placed under stress, if not killed outright, by
90 F water Even at 75 to 80 F, they may be un-
able to compete successfully with organisms for
which 75 to 80 F is a favorable temperature. Simi-
larly, the inhabitants of warmer waters are at a
competitive disadvantage in cool water.
Although in a rigorous climate, an animal can
endure the extremes of temperature at appropriate
seasons; it must be cooled gradually in the fall if it
is to become acclimatized to the cold water of
winter and warmed gradually in the spring if it is to
withstand summer heat Further, an organism
might be able to endure a high temperature of 92
or 95 F for a few hours, but it could not do so for
a period of days. Having the water change gradu-
ally with the season is important for other reasons.
an increasing or decreasing temperature often
serves as the trigger for spawning activities, meta-
morphosis, and migration Some fresh water orga-
nisms require that their eggs be chilled before they
will hatch properly.
In arriving at suitable temperature criteria, the
problem is to estimate how far the natural tem-
perature may be exceeded without adverse effects
Whatever requirements are suggested, a seasonal
cycle must be retained, the changes in temperature
must be gradual and the temperature reached must
not be so high or so low as to damage or alter the
composition of the desired population. In view of
the many variables, it seems obvious that no single
temperature requirement can be applied to the
United States as a whole, or even to one State; the
requirements must be closely related to each body
of water and its population To do this a tempera-
ture increment based on the natural water tempera-
ture is more appropriate than an unvarying num-
ber Using an increment requires, however, that
we have information on the natural temperature
conditions of the water in question, and the size of
the increment that can be tolerated by the desired
species
If any appreciable heat load is introduced into a
stream, it must be recognized that the species'
equilibrium will likely be shifted towards that char-
acteristic of a more southerly water
The seasonal temperature fluctuation normal to
the desired biota of a particular water must be
maintained Further, the sum of any increase in
temperature plus the natural peak temperature
should be of short duration and below the maxi-
mum temperature that is detrimental for such
periods
Recommendation for Warm Waters: To maintain a
well-rounded population of warm-water fishes, the fol-
lowing restrictions on temperature extremes and tem-
perature increases are recommended
(1) During any month of the year, heat should not
be added to a stream in excess of the amount that will
raise the temperature of the water (at the expected
minimum daily flow for that month) more than 5F
In lakes and reservoirs, the temperatures of the epi-
Iimnion, in those areas where important organisms are
most likely to be adversely affected, should not be
raised more than 3 F above that which existed before
the addition of heat of artificial origin The increase
should be based on the monthly average of the maxi-
mum daily temperature Unless a special study shows
that a discharge of a heated effluent into the hypolim-
nion or pumping water from the hypolimnion (for dis-
charging back into the same water body) will be desir-
able, such practice is not recommended
(2) The normal daily and seasonal temperature
variations that were present before the addition of heat,
due to other than natural causes, should be maintained
(3) The recommended maximum temperatures that
are not to be exceeded for various species of warm-
water fish are given in table III—1
-------
Recommendation for Cold Waters: Because of the
large number of trout and salmon waters which have
been destroyed, or made marginal or nonproductive,
the remaining trout and salmon waters must be pro-
tected if this resource is to be preserved.
(1) Inland trout streams, headwaters of salmon
streams, trout and salmon lakes and reservoirs, and the
hypolimmon of lakes and reservoirs containing sal-
monids should not be warmed. No heated effluents
should be discharged in the vicinity of spawning areas.
For other types and reaches of cold-water streams,
reservoirs, and lakes, the following restrictions are
recommended
(2) During any month of the year, heat should not
be added to a stream in excess of the amount that will
raise the temperature of the water more than 5 F
(based on the minimum expected flow for that month)
In lakes and reservoirs, the temperature of the epi-
limnion should not be raised more than 3 F by the ad-
dition of heat of artificial origin.
(3) The normal daily and seasonal temperature
fluctuations that existed before the addition of heat due
to other than natural causes should be maintained
(4) The recommended maximum temperatures that
are not to be exceeded for various species of cold water
fish are given in table Til—1
Note—For streams, total added heat (in BTU's)
might be specified as an allowable increase in tempera-
ture of the minimum daily flow expected for the month
or period in question This would allow addition of a
constant amount of heat throughout the period. Ap-
proached in this way for all periods of the year, sea-
sonal variation would be maintained. For lakes the
situation is more complex and cannot be specified in
simple terms.
TABLE 111-1
[Provisional maximum temperatures recommended as compati-
ble with the well-being of various species of fish and
their associated biota]
93 F. Growth of catfish, gar, white or yellow bass,
spotted bass, buffalo, carpsucker, threadfin shad,
and gizzard shad.
90 F- Growth of largemouth bass, drum, bluegill, and
crappie.
84 F: Growth of pike, perch, walleye, smallmouth bass,
and sauger.
80 F: Spawning and egg development of catfish,
buffalo, threadfin shad, and gizzard shad.
75 F: Spawning and egg development of largemouth
bass, white, yellow, and spotted bass
68 F: Growth or migration routes of salmonids and for
egg development of perch and smallmouth bass.
55 F: Spawning and egg development of salmon and
trout (other than lake trout)
48 F. Spawning and egg development of lake trout,
walleye, northern pike, sauger, and Atlantic
salmon.
Note—Recommended temperatures for other species, not
listed above, may be established if and when necessary in-
formation becomes available
Dissolved oxygen
Oxygen requirements of aquatic life have been
extensively studied. Excellent survey papers are
presented by Doudoroff (1957), Doudoroff and
Shumway (1967), Doudoroff and Warren
(1962), Ellis (1937), and Fry (1960). Much of
the work on temperature requirements also con-
siders oxygen and those bibliographies are equally
valuable.
Most of the research concerning oxygen require-
ments for freshwater organisms deals with fish, but
since fish depend upon other aquatic species for
food and would not remain in an area with an in-
adequate food supply, it seems reasonable to as-
sume that a requirement for fish would serve also
for the rest of the community The fish themselves
can be grouped into three categories according to
their temperature and oxygen requirements:
(1) the cold-water fish (e.g, salmon and trout),
(2) the warm-water game and pan fish (e.g., bass
and sunfish), and (3) the warm-water "coarse"
fish (e.g, carp and buffalo) The cold-water fish
seem to require higher oxygen concentrations than
the warm-water varieties. The reason is not known,
but it may be related to the fact that, for half
saturation, trout hemoglobin requires an oxygen
partial pressure three or four times that required
by carp hemoglobin under similar circumstances.
Warm-water game and pan fish seem to require a
higher concentration than the "coarse" fish, prob-
ably because the former are more active and
predatory
Relatively little of the research on the oxygen
requirements of fish in any of these three categories
is applicable to the problem of establishing oxygen
criteria because the endpoints have usually been
too crude. It is useless in the present context to
know how long an animal can resist death by as-
phyxiation at low dissolved oxygen concentrations;
we must know instead the oxygen concentration
that will permit an aquatic population to thrive. We
need data on the oxygen requirements for egg de-
velopment, for newly hatched larvae, for normal
growth and activity, and for completing all stages
of the reproductive cycle It is only recently that
experimental work has been undertaken on the
effects of oxygen concentration on these more
subtle endpoints As yet, only a few species have
been studied
One of the first signs that a fish is being affected
by a reduction of dissolved oxygen (DO) concen-
tration is an increase in the rate at which it venti-
lates its gills, a process accomplished in part by an
increase in the frequency of the opercular move-
ments The half dozen or so species (chiefly
warm-water game and pan fish) that have been
reported so far show a significant increase in fre-
quency as the DO concentration is reduced from
6 to 5 mg/1 (at about 72 F) and a greater increase
-------
from 5 to 4 mg/1. If the opercular rate is taken as
the criterion by which the adequacy of an oxygen
concentration is to be judged, then such evidence
as we have indicates 6 mg/1 as the required dis-
solved oxygen concentration Several field studies
have shown, however, that good and diversified
fish populations can occur in waters in which the
dissolved oxygen concentration is between 6 and
5 mg/1 in the summer, suggesting that a minimum
of 6 mg/1 is probably more stringent than neces-
sary for warm-water fishes Because the oxygen
content" of a body of water does not remain con-
stant, it follows that if the dissolved oxygen is
never less than 5 mg/1 it must be higher part of
the time. In some cases, good populations of
warm-water fish, including game and pan fishes,
occur in waters in which the dissolved oxygen may
be as low as 4 mg/1 for short periods. Three mg/1
is much too low, however, if normal growth and
activity are to be maintained. It has been reported
that the growth of young fish is slowed markedly if
the oxygen concentration falls to 3 mg/1 for part
of the day, even if it rises as high as 18 mg/1 at
other times. It is for such reasons as this that oxy-
gen criteria cannot be based on averages Five and
4 mg/1 are close to the borderline of oxygen con-
centrations that are tolerable for extended periods.
For a good population of game and pan fishes,
the concentration should be considerably more
than this.
The requirements of the different stages in the
life cycles of aquatic organisms must be taken into
account An oxygen concentration that can be
tolerated by an adult animal, with fully developed
respiratory apparatus, less intense metabolic re-
quirements, and the ability to move away from
adverse conditions, could easily be too low for eggs
and larval stages The eggs are especially vulner-
able to oxygen lack because they have to depend
upon oxygen diffusing into them at a rate sufficient
to maintain the developing embryos. Hatching,
too, is a critical time; recently hatched young need
relatively more oxygen than adults, but until they
become able to swim for themselves (unless they
are in flowing water) they must depend upon the
oxygen supply in the limited zone around them
These problems are not as great among species
that tend their eggs and young, suspend their eggs
from plants, or have pelagic eggs, as they are for
salmonids Salmonids bury their eggs in the gravel
of the stream away from the main flow of the water
thereby requiring a relatively high oxygen concen-
tration in the water that does reach them.
Recommendation: In view of the above considerations
and with the proviso that future research may make
revision necessary, the following environmental con-
ditions are considered essential for maintaining na-
tive populations of fish and other aquatic life
(1) For a diversified warm-water biota, including
game fish, daily DO concentration should be above
5 mg/1, assuming that there are normal seasonal and
daily variations above this concentration Under ex-
treme conditions, however, and with the same stipula-
tion for seasonal and daily fluctuations, the DO may
range between 5 mg/1 and 4 mg/1 for short periods of
time, provided that the water quality is favorable in
all other respects In stratified eutrophic and dystrophic
lakes, the DO requirements may not apply to the
hypolimnion In shallow unstratified lakes, they should
apply to the entire circulating water mass
These requirements should apply to all waters ex-
cept administratively established mixing zones In lakes,
such mixing zones must be restricted so as to limit the
effect on the biota In streams, there must be no blocks
to migration and there must be adequate and safe
passageways for migrating forms These zones of pas-
sage must be extensive enough so that the majority of
plankton and other drifting organisms are protected
(see section on zones of passage)
(2) For the cold water biota, it is desirable that DO
concentrations be at or near saturation This is espe-
cially important in spawning areas where DO levels
must not be below 7 mg/1 at any time For good growth
and the general well-being of trout, salmon, and other
species of (he biota, DO concentrations should not be
below 6 mg/1 Under extreme conditions they may
range between 6 and 5 mg/1 for short periods provided
that the water quality is favorable and normal daily
and seasonal fluctuations occur. In large streams that
have some stratification or that serve principally as mi-
gratory routes, DO levels may be as low as 5 mg/1 for
periods up to 6 hours, but should never be below 4
mg/1 at any lime or place
(3) DO levels in the hypolimnion of oligotrophy
small inland lakes and in large lakes should not be
lowered below 6 mg/1 at any time due to the addition
of oxygen-demanding wastes or other materials
Carbon dioxide
An excess of "free" carbon dioxide (as distin-
guished from that present as carbonate and bicar-
bonate) may have adverse effects on aquatic ani-
mals These effects range from avoidance reactions
and changes in respiratory movements at low con-
centrations, through interference with gas ex-
change at higher concentrations, to narcosis and
death if the concentration is increased further The
respiratory effects seem the most likely to be of
concern in the present connection
Since the carbon dioxide resulting from meta-
bolic processes leaves the organisms by diffusion,
an increase in external CO, concentration will
make it more difficult for it to diffuse out of the
organism. Thus, it begins to accumulate internally
The consequences of this internal accumulation
are best known for fish, but presumably the princi-
ples are the same for other organisms. As the CO:
accumulates, it depresses the blood pH, and this
-------
may have detrimental effects. Probably more im-
portant, however, is the fact that the greater the
blood C02 concentration, the less readily will the
animal's hemoglobin combine with dissolved oxy-
gen Thus the presence of much C02 raises the
minimum oxygen concentration which is tolerable
Since the combination of oxygen with hemoglobin
is inversely related to temperature, it is obvious
that C02, temperature, and oxygen are closely
related. Insufficient data are available at present
to permit us to state the greatest amount of dis-
solved carbon dioxide that all types of aquatic
organisms can tolerate and how these tolerable
concentrations vary with temperature and dis-
solved oxygen. Studies of the effect of C02 on
the oxygen requirements of several species of fish
indicate that C02 concentrations of the order of
25 mg/1 should not be detrimental, provided the
oxygen concentration and temperature are within
the recommended limits.
Recommendation: According to our rather meagre
knowledge of the subject, it is recommended that the
free CO» concentration should not exceed 25 mg/l
Oil
Oil slicks are barely visible at a concentration of
about 25 gal/sq mi (Amer. Petroleum Inst. 1949).
At 50 gal/sq mi, an oil film is 3.0xl0~6 inches
thick and is visible as a silvery sheen on the sur-
face. Sources of oil pollution are bilge and ballast
waters from ships, oil refinery wastes, industrial
plant wastes such as oil, grease, and fats from the
lubrication of machinery, reduction works, plants
manufacturing hydrogenated glycerides, free fatty
acids, and glycerine, rolling mills, county drains,
storm-water overflows, gasoline filling stations, and
bulk stations
Wiebe (1935) showed that direct contact by fish
(bass and bream) with crude oil resulted in death
caused by a film over the gill filaments. He also
demonstrated that crude oil contains a water-solu-
ble fraction that is very toxic to fish. Galtsoff, et al.
(1935) showed that crude oil contains substances
soluble in sea water that produce an anaesthetic
effect on the ciliated epithelium of the gills of
oysters Free oil and emulsions may act on the
epithelial surfaces of fish gills and interfere with
respiration. They may coat and destroy algae and
other plankton, thereby removing a source of fish
food, and when ingested by fish they may taint
their flesh.
Setteable oily substances may coat the bottom,
destroy benthic organisms, and interfere with
spawning areas. Oil may be absorbed quickly by
suspended matter, such as clay, and then due to
wind action or strong currents may be transported
over wide areas and deposited on the bottom far
from the source Even when deposited on the bot-
tom, oil continuously yields water-soluble sub-
stances that are toxic to aquatic life.
Films of oil on the surface may interfere with
reaeration and photosynthesis and prevent the
respiration of aquatic insects such as water boat-
men, backswimmers, the larvae and adults of
many species of aquatic beetles, and some species
of aquatic Diptera (flies). These insects surface
and carry oxygen bubbles beneath the surface by
means of special setae which can be adversely af-
fected by oil Berry (1951) reported that oil films
on the lower Detroit River are a constant threat to
waterfowl Oil is detrimental to waterfowl by de-
stroying the natural buoyancy and insulation of
their feathers
A number of observations made by various
authors in this country and abroad record the con-
centrations of oil m fresh water which are dele-
terious to different species For instance, penetra-
tion of motor oil into a fresh water reservoir
used for holding crayfish in Germany caused the
death of about 20,000 animals (Seydell, 1913)
It was established experimentally that crayfish
weighing from 35 to 38 g die in concentrations of
5 to 50 mg/1 within 18 to 60 hours Tests with two
species of fresh water fish, ruff (small European
perch), and whitefish (fam Coregonidae) showed
that concentrations of 4 to 16 mg/1 are lethal to
these species in 18 to 60 hours.
The toxicity of crude oil from various oil fields
in Russia varies depending on its chemical com-
position. The oil used by Veselov (1948) in the
studies of the pollution of Belaya River (a tribu-
tary in the Kama in European Russia) belongs to a
group of methano-aromatic oils with a high con-
tent of asphalt, tar compounds, and sulfur. It
contains little paraffin and considerable amounts
of benzene-ligroin. Small crucian carp (Carassius
carassius) 7-9 cm long were used as the bioassay
test animal This is considered to be a hardy fish
that easily withstands adverse conditions. The
water soluble fraction of oil was extracted by
shaking 15 ml of oil in 1 liter of water for 15
minutes. The oil film was removed by filtration.
Dissolved oxygen was controlled A total of 154
tests were performed using 242 fishes. The average
survival time was 17 days at the concentration of
0 4 ml/1 of oil but only 3 days at the concentration
of 4 ml/1 Further increase in concentration had no
appreciable effect on fish mortality.
Seydell (1913) stated that the toxicity of Rus-
sian oil is due to naphthemc acids, small quantities
of phenol, and volatile acids (Veselov, 1948).
-------
Cairns (1957) reports the following 96-hour TLm
values of naphthenic acid for bluegill sunfish
(Lepomis macrochirus)—5.6 mg/1; pulmonate
snail (Physa heterostropha)—6 1 to 7 5 mg/1 (in
soft water), and diatom (species not identified)—
41.8 to 43.4 mg/1 in soft water and 28 2 to 79 8
mg/1 in hard water. Naphthenic acid (cyclohexane
carboxylic acid) is extracted from petroleum and
is used in the manufacture of insecticides, paper,
and rubber
Chipman and Galtsoff (1949) report that crude
oil in concentrations as low as 0.3 mg/1 is ex-
tremely toxic to fresh water fish. Dorris, Gould,
and Jenkins (1960) made an intensive study of the
toxicity of oil refinery effluents to fathead minnows
in Oklahoma. By standard bioassay procedures,
they found that mortality varied between 3.1 per-
cent to 21.5 percent after 48 hours of exposure to
untreated effluents. They concluded that toxicity
rather than oxygen demand is the most important
effect of oil refinery effluents on receiving streams.
Pickering and Henderson (1966b) reported the
results of acute toxicity studies of several impor-
tant petrochemicals to fathead minnows, bluegills,
goldfish, and guppies in both soft water and hard
water Standard bioassay methods were used Be-
cause several of the compounds tested have low
solubility in water, stock solutions were prepared
by blending the calculated concentrations into 500
ml of water before addition to the test container
Where necessary, pure oxygen was supplied by
bubbling at a slow rate The petrochemicals tested
were benzene, chlorobenzcne, 0-chlorophenol, 3-
chloropropene, 0-cresol, cyclohexane, ethyl ben-
zene, isoprene, methyl methacrylate, phenol, 0-
phthalic anhydride, styrene, toluene, vinyl acetate,
and xylene These petrochemicals are similar in
their toxicities to fish, with 96-hour TLm values
ranging from 12 to 368 mg/1 Except for isoprene
and methyl methacrylate, which are less toxic,
values for all four species of fish for the other
petrochemicals ranged from 12 to 97 mg/1, a rela-
tively small variation In general, 0-chlorophenol
and 0-cresol are the most toxic and methyl meth-
acrylate and isoprene are the least toxic
Recommendation: In view of available data, it is con-
cluded that to provide suitable conditions for aquatic
life, oil and petrochemicals should not be added in
such quantities to the receiving waters that they will
(1) produce a visible color film on the surface, (2)
impart an oily odor to water or an oily taste to fish
and edible invertebrates, (3) coat the banks and bot-
tom of the water course or taint any of the associated
biota, or (4) become effective toxicants according to
the criteria recommended in the 'Toxicity" section
Turbidity
Turbidity is caused by the presence of suspended
matter such as clay, silt, finely divided organic
matter, bacteria, plankton, and other microscopic
oragnisms Turbidity is an expression of the optical
property of a sample of water which causes light to
be scattered and absorbed rather than transmitted
in straight lines through the sample. Excessive
turbidity reduces light penetration into the water
and, therefore, reduces photosynthesis by phyto-
plankton organisms, attached algae, and sub-
mersed vegetation
The Jackson candle turbidimeter (Standard
Methods for the Examination of Water and Waste-
water, 12th edition 1965) is the standard instru-
ment for making measurements of turbidity Field
determinations, however, are made with direct-
reading colorimeters calibrated for this test and the
results are expressed as Jackson turbidity units
(JTU)
Silt and sediment are particularly damaging to
gravel and rubble-type bottoms The sediment fills
the interstices between gravel and stones, thereby
eliminating the spawning grounds of fish and the
habitat of many aquatic insects and other inverte-
brate animals such as mollusks, crayfish, fresh
water shrimp, etc Tarzwell (1957) observed that
bottom organisms from a silted area averaged only
36 organisms/sq ft compared to 249/sq ft in a
non-silted area Smith (1940) reported that silting
reduced the bottom fauna of the Rogue River by
25 to 50 percent Observations in Oregon by Wag-
ner (1959) and Ziebell (1960) showed an 85-
percent decline in productivity of aquatic insect
populations below a gravel dragline operation.
Turbidities in the affected area were increased
from zero to 91 mg/1 and suspended solids from
2 mg/1 upstream to 103 mg/1 downstream
Buck (1956) investigated several farm ponds,
hatchery ponds, and reservoirs over a 2-year
period He observed that the maximum production
of 161 5 lb/acre occurred in farm ponds where
the average turbidity was less than 25 JTU. Be-
tween 25 and 100 JTU, fish yield dropped 41.7
percent to 94 lb/acre, and in muddy ponds, where
turbidity exceeded 100 JTU, the yield was only
29 3 lb/acre or 18 2 percent of clear ponds
Herbert and Merkens (1961), using a mixture
of kaolin and diatomaceous earth, demonstrated
that long-term exposure of rainbow trout to 100-
200 mg/1 could be harmful At 270 and 810 mg/1,
a high percentage of the fish died. Wallen (1951)
studied the effects of montmorillonite clay on 16
species of warm-water fish. Results are shown in
table III-2 It is shown that fish can tolerate high
-------
turbidities for short periods, a fortunate adaptation
for river species Fish productivity is ultimately
dependent upon plant life and a good bottom
fauna. There can be little of either above 200 JTU
if that turbidity is maintained continuously The
Aquatic Life Advisory Committee of the Ohio
River Valley Water Sanitation Commission
(ORSANCO) Second Progress Report (1956)
points out that fish withstand turbidities of 5,000
mg/l or more with no direct harmful results, but
the productivity of the bottom areas is very low
and the fish populations are small
TABLE III—2. Average Turbidities Found To Be
Fatal to Fish
Length of
Turbidity
Species
exposure (days)
(mg/l)
Large mouth bass
7 6
101,000
Pumpkin seed sunfish..
13
69,000
Channel catfish
9 3
85,000
Black bullhead
17
222,000
Golden shiner
7 1
166,000
Ellis (1937) summarized the results of 2,344
light penetration determinations made at 585 sta-
tions on streams throughout the United States The
determinations were made of the millionth inten-
sity depth (mid), which is the depth in milli-
meters of water of the given turbidity required to
screen out 99 9999 percent of the light entering at
the surface. A photoelectric apparatus described
by Ellis (1934b) was used and determinations
were made after filtering the water through bolting
silk
The turbidity of rivers varies widely in different
parts of the country Ellis (1937) defined clear
streams as those with amid of 5 00 to infinity,
cloudy streams, 4 90-1.00 meters, turbid, 0 99-
0 50; very turbid, 0 49-0 30; muddy, 0 29-0 15,
very muddy, 0 14-0 00 meters
In Mississippi River side channels and flowing
stream tributaries with good fish fauna, 4 percent
were clear, 11 percent cloudy, 3 percent were very
muddy In these waters, with medium, poor, or no
fish fauna 1 percent were clear, 18 percent cloudy,
11 percent turbid, 14 percent very turbid, 38 per-
cent muddy, and 18 percent very muddy
Based on 6,000 light penetration determinations
or inland streams, he concluded that, for good pro-
duction of fish and aquatic life, the silt load of
these streams should be reduced so that the mil-
lionth intensity depth would be greater than 5
meters.
Good farming practices can do a great deal to
prevent silt from reaching streams and lakes. Road
building and housing development projects, placer
mining, strip mining, coal and gravel washing, and
unprotected road cuts are important sources of
turbidity that can be reduced with planning, good
housekeeping, and regulation.
Natural turbidities within watersheds should be
determined For example, in some Western States
many streams have a turbidity below 25 JTU for
most of the year In those states, the water pollu-
tion control agency might specify that no wastes
should be discharged which would raise the tur-
bidity of the receiving water above 25 JTU.
From the above discussion it can be seen that
natural turbidity varies greatly in different parts of
the country
Recommendation. Turbidity in the receiving water
clue to a discharge should not exceed 50 JTU in warm-
water streams or 10 JTU in cold-water streams.
There should be no discharge to warm-water lakes
which will cause turbidities exceeding 25 Jackson Units
The turbidity of cold-water or oligotrophy lakes should
not exceed 10 units
Settleable solids
Settleable solids include both inorganic and or-
ganic materials The inorganic components include
sand, silt, and clay originating from such sources
as erosion, placer mining, mine tailing wastes, strip
mining, gravel washing, dusts from coal washeries,
loose soils from freshly plowed farm lands, high-
way, and building projects The organic fraction
includes such settleable materials as greases, oils,
tars, animal and vegetable fats, paper mill fibers,
synthetic plastic fibers, sawdust, hair, greases from
tanneries, and various settleable materials from city
sewers These solids may settle out rapidly and
bottom deposits are often a mixture of both inor-
ganic and organic solids They may adversely af-
fect fisheries by covering the bottom of the stream
or lake with a blanket of material that destroys the
bottom fauna or the spawning grounds of fish
Deposits containing organic materials may deplete
bottom oxygen supplies and produce hydrogen
sulfide, carbon dioxide, methane, or other noxious
gases
Some settleable solids may cause damage by
mechanical action
Water Quality Criteria for European Freshwater
Fish (European Inland Fisheries Advisory Com-
mission, 1964) discusses chemically inert solids
in waters that are otherwise satisfactory for the
maintenance of freshwater fisheries. It is indicated
that good or moderate fisheries can be maintained
in waters that normally contain 25 to 80 mg/l sus-
pended solids, but that the yield of fish might be
-------
lower than in waters containing 25 mg/1 or less
Waters normally containing 80 to 400 mg/1 sus-
pended solids are unlikely to support good fresh-
water fisheries
Recommendation: Since it is known that even minor
deposits of settleable materials inhibit the growth of
normal stream or lake flora and fauna, it is recom-
mended that no settleable materials be added to these
waters in quantities that adversely affect the natural
biota
Color
The color of water is attributed to substances
in solution after the suspensoids have been re-
moved It may be of organic or mineral origin
Organic sources are humic materials, peat, plank-
ton, rooted and floating aquatic plants, tannins,
etc Inorganic sources arc metallic substances such
as iron and manganese compounds and chemicals,
dyes, etc Many industries discharge materials that
contribute to the color of water Among them are
pulp and paper mills, textile mills, refineries,
manufacturers of chemicals and dyes, explosives,
nailworks, tanneries, etc
Standard Methods for the Examination of Water
and Wastewater, 12th edition (1965), describes
the standard platinum-cobalt method of determin-
ing color after centnfugation The unit of color
considered as standard is the color produced by
one mg/1 of platinum in water Results are ex-
pressed as units of color. Color in excess of 50
units may limit photosynthesis and have a dele-
tenous effect upon aquatic life, particularly phyto-
plankton, and the benthos
Water absorbs light differentially A layer of
distilled water 1 meter in thickness absorbs 53 per-
cent of the solar radiation It absorbs 30 percent of
the red-orange band (6,500 angstrom units) but
less than 5 percent of the blue (4,500 angstrom
units). These are the portions of the spectrum that
are absorbed and utilized to the greatest extent by
chlorophyll The band at 7,500 angstrom units is
over 90 percent absorbed
Natural waters absorb far more light The light
intensity at which the amount of oxygen produced
photosynthetically is balanced by the amount of
oxygen used for respiration in some submerged
vascular plants is 5% of full sunlight on clear
summer days It is estimated that 25 to 50 percent
of full sunlight is necessary for many green aquatic
plants to reach maximum photosynthesis The
ORSANCO committee observed that the 25-per-
ccnt level of solar radiation is not reached in many
of the larger streams and they considered it desira-
ble to restrict the addition of any substances that
reduce light penetration and hence limit the pri-
mary productivity of aquatic vegetation
Recommendation: For effective photosynthetic pro-
duction of oxygen, it has been found that at least
10 percent of incident light is required. Therefore,
10 percent of the incident light should reach the bottom
of any desired photosynthetic zone in which adequate
dissolved oxygen levels are to be maintained
Floating materials
Floating materials include sawdust, peelings
and other cannery wastes, hair and fatty materials
from tanneries, wood fibers, containers, scums, oil,
garbage, floating materials from untreated munic-
ipal and industrial wastes, tars and greases, and
precipitated chemicals
Wastes from paper mills, vinegar plants, cane
mills, and other industries may contribute nutrients
or produce conditions in streams that foster the
growth of Sphaerotilus (Chlamydobacteriales)1 or
similar iron or sulfur bacteria. These floating
growths not only clog fishermen's nets, but also
smother out the spawning grounds and habitat of
all forms of aquatic life
Recommendation: All such floating and settleable sub-
stances should be excluded from streams and lakes
Tainting substances
Among the materials that are responsible for
objectionable tastes in fish are hydrocarbons,
phenolic compounds, sodium pentachlorophenate
(used for slime control in cooling towers), coal
tar wastes, gas wastes, sewage containing phenols,
coal-coking wastes, outboard motor exhaust
wastes, and petroleum refinery wastes Kraft paper
mill wastes, sulfides, mercaptans, turpentine,
wastes from synthetic rubber and explosives fac-
tories, algae, resins and resin acids also contribute
to objectional tastes in fish Twenty gallons per
acre of kerosene or diescl fuel will produce an off-
flavor in bass and bluegills which persists for 4 to
6 weeks The Aquatic Life Advisory Committee
of ORSANCO in its Third Progress Report
(1960), lists the concentrations (table III—3) of
phenolic substances that cause taste and odor
Albersmeyer and Erichscn (1959) found that car-
bolated oil and light oil, both dephenolated, im-
part a taste to fish flesh more pronounced than
that caused by naphthalene and methyl naphtha-
lene They concluded that the hydrocarbons are
more responsible for tastes in fish flesh than the
phenolic compounds. Boetius (1954) found that
chlorophenol could produce unpleasant flavor in
fish at a concentration of only 0 0001 mg/1.
-------
TABLE III—3. Concentration of Phenolic Com-
pounds That Cause Tainting of Fish Flesh
After Bandt (1955) page 77 (except for phenol)
Concentration
Compound or affecting taste
waste and odor (mg/l) Fish tested
Pure compounds:
Phenol 15 to 25 Trout, carp, tench,
chub, eel, min-
now, perch, blue-
gill, pike, gold-
fish
Cresols _ ...10 .. Tench, carp, eel,
trout, minnow
Xylenols 1 to 5 Roach, perch, carp
Pyrocatechol 2 to 5 Perch, carp, roach.
Pyrogallol 20 to 30 Roach, carp
P-Quinone 0 5 Carp, tench, roach
Pyridine 5 ... .Roach, carp
Naphthalene 1 Roach
Alpha NaphthoL.O 5 Roach, carp.
Quinoline 0 5 to 1 0 do
Chlorophenol ...0 1-
Mixed phenolic wastes:
Coal-coking
wastes 0 02 to 0 1 Freshwater fish.
Coal-tar wastes..0 1 do
Phenols in
polluted river._0 02 to 0 15... Minnows
Sewage contain-
ing phenols ..0 1 Freshwater fish.
A preliminary laboratory study (English, Mc-
Dermott, and Henderson, 1963) shows that out-
board motor exhaust damages the quality of water
in several ways, the most noticeable of which is
causing unpleasant taste and odor in the water and
off-flavoring of fish flesh. A later field study, Eng-
lish et al. (1963a, b) and Surber et al. (1965)
determined the threshold level of tainting of fish in
pond and lake waters to be about 2.6 gal/acre-foot
of fuel, accumulating over a 2-month period. The
gasoline used was regular grade and the lubricating
oil (y> pint/gal) was a popular brand of packaged
outboard motor oil
Recommendation: Materials that impart odor or taste
to fish flesh or other freshwater edible products such as
crayfish, clams, prawns, etc, should not be allowed to
enter receiving waters at levels that produce tainting
Where it seems probable that a discharge may result in
tainting of edible aquatic products, bioassays and laste
panels are suggested for determining whether tainting
is likely
Radioactive materials in fresh
and marine waters
Ionizing radiation, when absorbed in living
tissue in quantities substantially above that of nat-
ural background, is recognized as injurious It is
necessary, therefore, to prevent excessive levels of
radiation from reaching any organism we wish to
preserve, be it human, fish, or invertebrate. Beyond
the obvious fact that they emit ionizing radia-
tion, radioactive wastes are similar in many re-
spects to other chemical wastes. Man's senses can-
not detect radiation unless it is present in massive
amounts Radiation can be detected, however, by
means of electronic instruments and quantities
present at very low levels in the environment can
be measured with remarkable accuracy. Because
of the potential danger, the disposal of radioactive
materials has been well planned and controlled
Injuries and loss of life from disposal of radioactive
materials or from accidents involving these mate-
rials have been minimal Four factors have con-
tributed to this safety record. (1) scientists and
legislators were aware of the dangers associated
with the release of radioactive materials into the
environment prior to the need for disposal; (2) re-
search has progressed to protect man against ra-
diation effects and levels of radiation that could be
released, (3) as knowledge of nuclear energy in-
creased, standards were developed for handling,
shipping, and disposing of radioactive substances;
and (4) an extensive monitoring program was in-1
augurated and has been functioning for years.
Upon introduction into an aquatic environment,
radioactive wastes can. (1) remain in solution
or in suspension, (2) precipitate and settle to
the bottom, or (3) be taken up by plants and ani-
mals. Immediately upon introduction of radioac-
tive materials into the water, certain factors inter-
act to dilute and disperse these materials, while
simultaneously other factors tend to concentrate
the radioactivity Among those factors that dilute
and disperse radioactivity are currents, turbulent
diffusion, isotopic dilution, and biological trans-
port Radioactivity is concentrated biologically by
uptake directly from the water and passage
through food webs, chemically and physically by
adsorption, ion exchange, coprecipitation, floccula-
tion, and sedimentation
Radioactive wastes in the aquatic environment
may be cycled through water, sediment, and the
biota. Each radionuclide tends to take a charac-
teristic route and has its own rate of movement
from component to component prior to coming to
rest in a temporary reservoir, one of the three
components of the ecosystem Isotopes can move
from the water to the sediments or to the biota. In
effect, the sediments and biota compete for the
isotopes in the water Even though in some in-
stances sediments are initially successful in remov-
ing large quantities of radionuclides from the
water, and thus preventing their immediate uptake
by the biota, this sediment-associated radioactivity
-------
may latei affect many benthic species by exposing
them to radiation Also, any radioactivity leached
from the sediments back to the water again be-
comes available for uptake by the biota Even be-
fore the radioactivity is leached from the sediment,
it may become available to the biota due to a
variation in the strength of the bonds between the
different radionuclides and the sediment particles
Loosely bound radionuclides can be "stripped"
from particles of sediment and utilized by bottom-
feeding organisms.
Plants and animals, to be of any significance in
the cycling of radionuclides in the aquatic environ-
ment, must accumulate the radionuclide, retain it,
be eaten by another organism, and be digestible
However, even if an organism accumulates and
retains a radionuclide and is not eaten before it
dies, the radionuclide will enter the "biological
cycle" through organisms that decompose the dead
organic material into its elemental components
Plants and animals that become radioactive in this
biological cycle can pose a health hazard when
eaten by man
Aquatic life may receive radiation from radio-
nuclides present in the water and substrate and
also fiom radionuclides that may accumulate
within their tissues Humans can acquire radionu-
clides via many pathways, but among the most
important are drinking water or edible fish and
shellfish that have concentrated nuclides from the
water In order to prevent unacceptable doses of
radiation from reaching humans, fish, and other
important organisms, the concentrations of radio-
nuclides in water, both fresh and marine, must be
restricted
The effects of radiation on organisms have been
the subject of intense investigation for many years
Careful consideration of pertinent portions of the
vast amount of available information by such or-
ganizations as the International Commission on
Radiological Protection (ICRP), the National
Committee on Radiation Protection and Measure-
ments (NCRP), and the Federal Radiation Coun-
cil (FRC) has resulted in recommendations on the
maximum doses of radiation that people may be
allowed to receive under various circumstances
(U S. Department of Commerce, 1963) The rec-
ommended levels for the general public are sub-
stantially more conservative than those for persons
who work with radiation sources or radionuclides,
but in both cases the recommended levels assume
that the exposure will be sustained essentially
throughout the life or period of employment of the
person
The ICRP and NCRP have calculated the quan-
tities of individual radionuclides that a person can
ingest each day without accumulating levels in
various body organs that deliver radiation doses
in excess of the recommended limits These quanti-
ties contained in the volume of water ingested
daily (2 2 liters) are referred to as "maximum per-
missible concentrations (MPC) in water" The
FRC, recognizing that people may jngest radio-
nuclides from foods and other sources as well as
from drinking water, has provided guidance on
the basis of transient rates of intake from all
sources, but only for a few nuclides (radium-226,
iodine-131, strontium-90, and strontium-89).
The PHS Drinkihg Water Standards (US-
DHEW, 1962) are responsive to the recom-
mendations of the FRC, ICRP, and NCRP, and
provide appropriate protection against unaccept-
able radiation dose levels to people where drinking
water is the only significant source of exposure
above natural background. Where fish or other
fresh or marine products that have accumulated
radioactive materials are used as food by humans,
the concentrations of the nuclides in the water
must be further restricted to provide assurance
that the total intake of radionuclides from all
sources will not exceed the recommended levels.
The radiation dose received by fish and other
aquatic forms will be greater than that received by
people who drink the water or eat the fish Even
so, this does not place the fish in risk of suffering
radiation damage The radiation protection guides
for people have been established with prudence,
for continued exposure over a normal life span,
and with appropriate risk (safety) factors Virtu-
ally all of the available evidence shows that the
concentrations of radionuclides in fish and shell-
fish that would limit their use as food are substan-
tially below the concentrations that would injure
the organisms from radiation Therefore, at this
time there appears to be no need for establishing
separate criteria for radioactive materials in water
beyond those needed to limit the intake to humans.
Recommendation. (1) No radioactive materials
should be present in receiving waters as a consequence
of the failure of an installation to exercise practical and
economical controls to minimize releases This recom-
mendation is responsive to the recommendations of the
FRC that "There can be no single permissible or ac-
ceptable level of exposure without regard to the reason
for permitting the exposure It should be general prac-
tice to reduce exposure to radiation, and positive effort
should be carried out to fulfill the sense of these recom-
mendations It is basic that exposure to radiation should
result from a real determination of its necessity "
(2) No radionuclide or mixture of radionuclides
should be present at concentrations greater than those
specified in the PHS Drinking Water standards
(USDHEW, 1962) This recommendation assures that
people will receive no more than acceptable amounts
-------
of radioactive materials from aquatic sources and that
fish living in the water will not receive an injurious
dose of radiation.
(3) The concentrations of radioactive materials
present in fresh, estuarine, and marine waters should be
less than those that would require restrictions on the
use of organisms harvested from the area in order to
meet the Radiation Protection Guides recommended
by the Federal Radiation Council
This recommendation assures that fish and other
fresh water and marine organisms will not accu-
mulate radionuclides to levels that would make
them unacceptable for human food. It also limits
the radiation dose that the organisms would receive
from internally deposited nuclides to levels below
those that may be injurious Some workers (Car-
ritt, 1959; Isaacs, 1962; Pritchard, 1959) have
recommended "maximum permissible levels for
sea water" based on various assumptions of dis-
persion, uptake by marine organisms, and the use
of the organisms as food by people While these
recommendations are most useful as a first ap-
proximation in predicting safe rates of discharge
of radioactive wastes, their applicability as water
quality criteria is limited and they are not intended
for use in fresh or estuarine waters where the con-
centrations of a great variety of chemical elements
vary widely. Because it is not practical to general-
ize on the extent to which many of the important
radionuclides will be concentrated by fresh water
and marine forms, nor on the extent to which these
organisms will be used for food by people, no at-
tempt is made here to specify MPC for either sea
water or fresh water in reference to uptake by the
organisms Rather, each case requires a separate
evaluation that takes into account the peculiar fea-
tures of the region. Such an evaluation should be
approved by an agency of the State or Federal
Government in each instance of radioactive con-
tamination in the environment. In each particular
instance of contamination, the organisms present,
the extent to which these organisms concentrate
the radionuclides, and the extent to which man
uses the organisms as food must be determined, as
well as the rates of release of radionuclides must be
based on this information
Plant nutrients and nuisance
organisms
All terrestriat biological processes plus the ma-
jority of man's activities ultimately result in waste
products in various stages of decomposition A
portion of these sooner or later enter surface fresh-
waters These waste products include a rather
abundant amount of plant nutrients such as nitro-
gen, phosphorus, carbon, and other elements
Subsequently, these plant nutrients are incorpo-
rated into organic matter by aquatic plants.
Surface water areas are like land areas in that
some type of vegetation will occupy any suitable
habitat. Thus, the more abundant the nutrient sup-
ply, the more dense the vegetation, provided other
environmental factors are favorable. In the aquatic
habitat, these growths may be bacteria, aquatic
fungi, phytoplankton, filamentous algae, sub-
mersed, emersed, floating, and marginal water
plants. Practically all aquatic plants may be de-
sirable at one time or another and in one habitat
or another. However, when they become too dense
or interfere with other uses of the water or of the
aquatic habitat, they become nuisance growths.
Some sheath-forming bacteria are the primary
nuisance-type growths in rivers, lakes, and ponds.
A notable problem associated with this group oc-
curs in areas subjected to organic enrichment. The
most common offenders belong to the genus Sphae-
rotilus These bacteria are prevalent in areas re-
ceiving raw domestic sewage, improperly stabilized
paper pulp effluents or effluents containing simple
sugars. The growths they produce interfere with
fishing by fouling lines, clogging nets, and gener-
ally creating unsightly conditions in the infested
area Their metabolic demands while they are liv-
ing and their decomposition after death impose a
high BOD load on the stream and can severely
deplete the dissolved oxygen. It has been suggested
that large populations of Sphaerotilus render
the habitat noxious to animals and hence its
presence may actively exclude desirable fish and
invertebrates
The freshwater algae are diverse in shape,
color, size, and habitat A description of all spe-
cies of algae would be as comprehensive as writing
about all land plants, mosses, ferns, fungi, and
seed plants
They may be free floating (planktonic) or they
may grow attached to the substrate (benthic or
epiphytic types). They may be macroscopic or
microscopic and are single-celled, colonial, or fila-
mentous They are the basic link in the conversion
of inorganic constituents in water into organic
matter When present in sufficient numbers, these
plants impart a green, yellow, red, or black color
to the water They may also congregate at or
near the water surface and form so-called "water-
bloom" or "scum."
A major beneficial role of algae is the removal
of carbon dioxide from the water by photosyn-
thesis during daylight and the production of oxy-
gen Algae, like other organisms, continually
-------
respire and produce carbon dioxide The amount
of oxygen produced during active photosynthesis
is many times the amount of carbon dioxide re-
leased during the night or on cloudy days when
photosynthesis is inhibited or stopped.
Limited concentrations of algae are not trouble-
some in surface waters; however, overproduction
of various species is considered undesirable for
many water uses A relatively abundant growth of
planktonic algae in waters 3 feet or deeper will
shade the bottom muds sufficiently to prevent
germination of seeds and halt the growth of prac-
tically all rooted submersed and emersed aquatics,
thus removing an important source of food for
ducks and other water fowl
Some blue-green algae, many green algae, and
some diatoms produce odors and scums that make
waters less desirable for swimming Dense growths
of such planktonic algae may limit photosynthetic
activity to a layer only a few inches beneath the
surface of the water. Under certain conditions, the
populations of algae may die and their decomposi-
tion will deplete dissolved oxygen in the entire
body of water. Certain sensitive people are allergic
to many species of planktonic algae blooming in
waters used for swimming
It is claimed that some species of algae cause
gastric disturbances in humans who consume such
infested waters Several species of blue-green algae
produce, under certain conditions, toxic organic
substances that kill fish, birds, and domestic ani-
mals Some of the genera that contain species
which may produce toxins are Anabaena, Ana-
cystis, Aphanizomenon, Coleolosphaertum, Gloeo-
trichia, Microcystis, Nodularia, and Nostoc Some
species of Chlorella, a green alga, also are toxic
Various species of single, as well as branched
filamentous forms of algae, grow in both cool
and warm weather and when they become over-
abundant are generally considered to be a nui-
sance in whatever body of water they occur. Most
species of these algae are generally distributed over
the United States
Many forms of plankton and filamentous algae
clog sand filters in water treatment plants, produce
undesirable tastes and odors in drinking water,
and secrete oily substances that interfere with do-
mestic use and manufacturing processes Some
algae cause water to foam during heating as well
as metal corrosion and the clogging of screens,
filters, and piping Algae also coat cooling towers
and condensers causing these units to become in-
effective. In Lake Superior, complaints have been
made that diatoms such as Tabellaria, Synedra,
Cymbella, and Fragilaria, and the chrysophyte,
Dmobryon, may be the cause of slimes on fishnets
Filamentous algae may interfere with the opera-
tion of irrigation systems by clogging ditches,
wires, and screens and thus seriously impede the
flow of water. Filamentous algae in ponds, lakes,
and reservoirs may cause depletion of naturally
occurring and added nutrients that could other-
wise be used to produce unicellular algae that are
more commonly used as food by fish Dense
growths of filamentous algae may reduce the total
fish production and seriously interfere with har-
vesting the fish either by hook and line fishing,
seining, or draining. Such growths can also cause
overpopulation, resulting in stunting and the pres-
ence of large numbers of small fish Under cer-
tain conditions, growths of filamentous algae on
pond or lake bottoms become so dense that they
eliminate spawning areas of fish and possibly inter-
fere with the production of invertebrate fish food.
Submersed plants are those which produce ajl
or most of their vegetative growth beneath the
water surface. In many instances these plants have
an underwater leaf form, a totally different floating
or emersed leaf form, and flowers on an aerial
stalk Abundant growth of these weeds is depend-
ent upon depth and turbidity of water, and sub-
stratum For most submersed plants in clear water,
8 to 10 feet is the maximum depth for growth in
clear water as they must receive sufficient light
for photosynthesis when they are seedlings Most
of these submersed aquatic plants appear capable
of absorbing nutrients as well as herbicides through
either their roots or vegetative parts
Emersed plants are rooted in bottom muds and
produce a majority of their leaves and flowers at
or above the water surface Some species have
leaves that are flat and float entirely upon the
water surface Other species have leaves that are
saucer-shaped or whose margins are irregular or
fluted The latter types of leaves do not float
entirely upon the water surface
Marginal plants are probably the most widely
distributed of the rooted aquatic plants Members
of this group are varied in size, shape, and prefer-
ence of habitat Many species are adapted for
growth from moist soils into water up to 2 feet
deep or more Other species are limited to moist
soil or entirely to a watery habitat
There are some species of floating plants that are
rather limited in their distribution while others
are widespread throughout the world Plants in
this group have true roots and leaves, but instead
of being anchored in the soil they float about on
the water surface Buoyancy of the plant is ac-
complished through modification of the leaf (in-
cluding covering of the leaf surface) and leaf
petiole Most species have well-developed root
-------
systems which collect nutrients from the water.
Species designated as weeds are not necessarily
such in all places and at all times. For example,
many submersed, floating, and emersed plants that
normally interfere with boating, swimming, and
fishing are regarded as desirable growths in water-
fowl refuge areas Rooted plants with floating
leaves, such as water lillies and watershield, and
those that float upon the surface, such as water
hyacinth, elodea, parrotweed, alligatorweed, and
duckweed, are considered highly objectionable for
many water uses. In clear water areas, however,
where artificial or natural fertilization is moderate,
the removal of these surface-shading plants may
permit sunlight to penetrate to the bottom muds
and submersed plants soon will occupy these
waters These submersed plants generally are more
objectionable in an area than the original surface-
covering plants.
Most emersed, marginal, and a few submersed
plants and filamentous algae produce growths that
provide a suitable habitat for the development of
anopheline and other pest-type mosquitoes as well
as a hiding place for snakes. They are excellent
habitats for damselflies and some aquatic beetles
Most rooted and floating aquatic plants can
seriously interfere with navigation of small rec-
reational craft and large commercial boats in in-
fested areas. Such problems are prevalent in
intercoastal waterways and in some streams in
the Gulf States area Water shortages due to con-
sumption by undesirable aquatic plants or reduc-
tion in carrying capacity of an irrigation or drain-
age canal through excessive vegetation can result
m decreased crop quality, yield, or even crop
failure.
Submersed and emersed weeds consume nutri-
ents, either available or added, that could other-
wise be used to grow desirable planktonic algae
in impounded waters Thus, the presence of ex-
cessive rooted plants may reduce total fish pro-
duction in the infested body of water Extensive
growth of weeds provides dense cover that allows
the survival of excessive numbers of fish resulting
in overcrowding and stunting as well as interfer-
ing with harvesting the fish by hook and line or
other methods There is evidence that rank growths
of submersed, emersed, or floating weeds may de-
plete the dissolved oxygen supply in shallower
water and that fish tend to leave these areas if
theie are open-water areas available of better
quality Although they carry on the process of
photosynthesis, their multicellular structure often
makes them less effective in re-oxygenating the
water
All the elements essential for plant growth arc
yet to be determined Some of the elements known
to be important are nitrogen, phosphorus, potas-
sium, magnesium, calcium, manganese, iron, sili-
con (for diatoms), sulfur (as sulfates), oxygen,
and carbon. In many habitats, abundance of the
first two elements, N (nitrogen) and P (phos-
phorus), promotes vegetative production if other
conditions for growth are favorable. Most algae
also require some simple organics, such as amino
acids and vitamins, and many trace elements, such
as manganese and copper Not only are the various
factors important, but their relative abundance and
combined affect can be of even greater importance.
Limited laboratory studies made to date indicate
that different species of algae have somewhat dif-
ferent phosphorus requirements with the range
of available phosphorus usually falling between
0 01 and 0 05 mg/1 as phosphorus. At these levels,
when other conditions are favorable, blooms may
be expected. As has been pointed out by the Sub-
committee on Water Quality Criteria for Public
Water Supply, the total phosphorus is of outstand-
ing importance While there is no set relationship
between total and available phosphorus (because
the ratio varies with season, temperature, and plant
growth), the total phosphorus is governing as it
is the reservoir that supplies the available phos-
phorus It is believed that allowable total phos-
phorus depends upon a variety of factors; e g,
type of water, character of bottom soil, turbidity,
temperature, and especially desired water use. Al-
lowable amounts of total phosphorus will vary,
but in general it is believed that a desirable guide-
line is 100 ng/l for rivers and 50 ng/\ where
streams enter lakes or reservoirs (recommended
by the Public Water Supply Subcommittee)
The nitrogen-phosphorus ratio is also of impor-
tance The ratio varies with the water, season, tem-
perature, and geological formation, and may range
from 1 or 2 1 to 100 1 In natural waters, the
ratio is often very near 10 1, and this appears to
be a good guideline for indicating normal condi-
tions
The major sources of nitrogen entering fresh
waters are atmospheric (approximately 5 lbs/
acre/year), (Hutchinsen, 1957), domestic sewage
effluents, animal and plant processing wastes, ani-
mal manure, fertilizer and chemical manufacturing
spillage, various types of industrial effluents, and
agricultural runoff
The major sources of phosphorous entering
fresh waters are domestic sewage effluents (in-
cluding detergents), animal and plant processing
wastes, fertilizer and chemical manufacturing spill-
age, various industrial effluents, and, to a limited
extent, erosion materials in agricultural runoff.
-------
Phosphorous entering an ecosystem may produce
a high oxygen demand. It has been pointed out
that 1 milligram of phosphorous from an organic
source demands about 160 milligrams of oxygen
in a single pass through the phosphorus cycle to
complete oxidation. Thus the oxidation of organic
matter, the growth of which has been induced by
adding phosphorus, may bring about a great reduc-
tion of oxygen in a lake or stream
Dissolved carbon in the form of simple organic
compounds can be utilized by many kinds of algae.
These types of carbon compounds are also used
direcdy as a source of food by many animals
Varying amounts of simple organic compounds
containing carbon are found in sewage and several
types of industrial wastes. Other more complex
forms of organic carbon can be utilized by bac-
teria The most common nuisance growth that
becomes very abundant in the presence of very
small amounts of carbon is Sphaerotilus. Patrick
(unpublished data) has shown that the addition
of 0 05-0.1 mg/1 of glucose, without changing
other ecological conditions, may produce nuisance
growths.
Knowledge of the nutrient requirements of fungi,
phytoplankton, and filamentous algae is more ex-
tensive than for rooted aquatic plants Laboratory
data on nutrient requirements must be used with
caution, however, because the maintenance of
most long-term cultures has required that extracts
of soil be incorporated into the inorganic culture
medium Analyses of field grown algae have indi-
cated a wide divergence in elemental composition
among various species and among the same species
from different localities Excessive growths often
seen to be triggered by small amounts of so-called
minor or trace elements and vitamins, particularly
Bi2
One of the most obvious effects of increases or
imbalances in nutrients is the change in the kinds
and abundance of species composing the algal
flora Historical studies of Lake Erie show a
change from an Aslerionella dominance in the
spring and a Synedra dominance in the fall of
1920 to a Melosira dominance in the spring and
a Melosira, Anabaena, Oscdlatoria dominance in
the fall of 1962. Between 1919 and 1934, the
number of cells per ml, with two exceptions, al-
ways were less than 4,000/ml Since 1934, the
cell count, with one exception, has always been
greater than 4,000/ml. In 1944, it reached 11,032
cells/ml. It should be pointed out that blue-green
algae are a poor source of food for most aquatic
life.
Benthic forms also indicate the increase in nu-
trients in an ecosystem Various species of Clado-
phora become abundant in lakes and rivers when
nutrients are abundant and replace the original
diverse benthic flora.
This demand for a wide variety of nutrients
is also characteristic of many of the rooted
aquatic plants. Their affinity for numerous metals,
however, does not appear to be comparable to
that of the algae
Extensive data exist on the concentration of
nitrogen and phosphorus in fresh waters through-
out the United States. (Allee, et al., 1949; Ellis,
1940; Engelbrecht and Morgan, 1961; Juday,
et al, 1927; Lackey, 1945; USDHEW, 1962a)
In evaluating these data, it must be remembered
that algae and most other aquatic plants are capa-
ble of utilizing any available N and/or P in a very
short time providing other growth conditions are
favorable. Thus, analyses of filtered water would
not provide an evaluation of all elements existing
in the original water sample. A more meaningful
figure would result if all materials in an original
water sample were digested and then analyzed.
Often, the dissolved or available phosphorus may
be very low, while the total amount in the orga-
nisms and organic matter may be quite large. Not
only does this determination of total phosphorous
give a better estimate of the existing nutrient load
of an area, but it also provides an index to the
potential release that would occur if these plants
should all die within a short period of time
This information would also point out the fact
that in many freshwaters, various species of rooted
aquatic plants are excellent receptors for this nu-
trient load Their use in effluent treatment might
be one of the cheaper waste-treatment procedures
The chemical composition of several species of
plants is given in table III-4, Indications are that
the N-P content of freshwaters in the United
States is quite varied, and their presence in fairly
large amounts may or may not produce algae
blooms.
It must be remembered that factors other than
plant nutrients also are operative in the establish-
ment and maintenance of aquatic plant growths.
There must be sufficient light reaching the plant
for photosynthesis to occur. If turbidity from
muds, dyes, other materials, or even phyto-
plankton is too great, plants at lower depths can-
not grow These same plants, however, if estab-
lished in an area, can trap large amounts of
intermittent silt and other materials and clear the
waters for downstream uses
Another factor that might be operative in pre-
venting aquatic plant growth would be the lack
of free C02 and bicarbonate ions in a particular
-------
aquatic environment. Certainly in an area where
the pH is high (9.5 or above) or low (below 5 5),
productivity would not reach high levels due to
a lack of sufficient bicarbonates.
Temperature also is an important factor in de-
lermimng the amount of growth. For each species,
there is an optimum range in which the greatest
growth occurs.
Wave action on large expanses of water may
also be a factor in regulating all types of aquatic
plant growths. This appears contradictory to the
concept that winds cause mixing of surface and
bottom waters, thereby renewing plant nutrients
in the euphotic zone. However, in certain lakes
and reservoirs, wind-induced waves and currents
mechanically agitate bottom materials and waters
to an extent that interferes with the production of
phytoplankton and rooted aquatic plants
Various workers have discussed the concentra-
tions of nitrogen and phosphorus that are needed
lor an algal bloom. Sawyer (1947) suggests that
a concentration of at least 15 fig/1 of phosphorus
is necessary for growth. Hutchinson (1957) states
that Asterionella can take up phosphorus from
where it is present at less than one pg/1. As a re-
sult of the study of 17 Wisconsin lakes, Mac-
kenthun (1965) cites results indicating that in-
organic nitrogen at 0.30 mg/1 and inorganic
phosphorus at 0.01 mg/1, at the start of an active
growing season, subsequently permitted algal
blooms. As yet, there is no definite information on
the amount of wastes that will produce predictable
harmful effects in a lake There are indicators,
however, of developing or potentially undesirable
conditions
There are several conditions, analyses, or meas-
ures that will indicate eutrophication and dystro-
phication Since these parameters are not infallible,
it is well to use them in combinations Conditions
indicative of organic enrichment are:
(1) A slow overall decrease year after year in
the dissolved oxygen in the hypolimnion
as indicated by determinations made a
short time before the fall overturn and an
increase in anaerobic areas in the lower
portion of the hypolimnion
TABLE 111-4. Chemical Composition of Some Algae From Ponds and Lakes in Southeastern
United States1
Giant
Analysis
Cham
Ptthophora
Spirogyra
Spirogyra
Rhizoctonium
Oedogonium
Mougeotis
Anabaena
Ash percent ...
... 43 4
27 77
13 06
13 86
17 36
12 69
14 54
5.19
C percent
... 29 3
35 38
42 40
41 16
39.10
40 84
40 74
49 70
N percent
... 2 46
2 57
3 01
2 35
3 46
264
1 77
9 43
P percent
... 0 25
0 30
0 20
0 23
0 43
0 08
0 25
0 77
S percent ...
... 0 55
1 42
0 27
0 24
0 27
0 15
0 36
0 53
Ca percent .. ..
... 8 03
3 82
0 57
084
0 52
044
1 68
0 36
Mg percent
... 0 92
0 20
0 45
0.30
0 21
0 16
0 57
0 42
K percent
... 2 35
3 06
0 92
0 99
1 90
3 03
1 20
1 20
Na percent
... 0 13
0 07
1 42
1 43
009
0 06
0 49
0 18
Fe mg/1
... 2,520
2,836
1,368
1,793
1,820
1,645
60
80
Mn mg/1
... 2,926
829
1,641
1,658
1,687
1,729
1,080
800
Zn mg/t __
89
29
72
46
89
119
520
Cu mg/1 -
19
23
47
34
75
75
143
70
B mg/1
6 7
65
4 2
43
1 8
8 1
8
...
Analysis Cladophora Euglena Hydrodictyon Microcystis Lyngbys Nltella Aphanlzomenon
Ash percer
C percent
N percent
P percent
S percent
Ca percent
Mg perceni
K percent
Na perceni
Zn mg/1
. 23 38
4 12
17 94
62
17 20
19 11
7 21
. 35 27
48 14
39 96
46 46
40 23
38 43
47.65
2 30
5 14
3 87
8 08
5 01
2 70
8 57
. 0 56
0 67
0 24
0 68
031
0 23
1 17
1 58
0 19
1 41
0 27
0 28
0 34
1 18
. 1 69
0.05
0 69
0 53
0 45
1 89
0 73
. 0 23
0 07
0 17
0 17
0 14
0 95
0 21
. 608
0 34
4 21
0 79
0 42
3.73
0.68
. 0 18
0 02
0 38
004
0 06
0 28
0.19
. 1,040
240
1,373
2,751
5,230
2,388
167
. 2,300
1,545
1,963
322
3,866
2,180
833
10
73
129
48
171
240
120
190
290
114
37
101
39
187
846
38
...
36
112
98
...
1 Lawrence (personal communication)
-------
(2) An increase in dissolved solids—especially
nutrient materials such as nitrogen, phos-
phorus, and simple carbohydrates.
(3) An increase in suspended solids—espe-
cially organic materials.
(4) A shift from a diatom-dominated plankton
population to one dominated by blue-green
and/or green algae, associated with in-
creases in amounts and changes in relative
abundance of nutrients
(5) A steady though slow decrease in light
penetration.
(6) An increase in organic materials and nu-
trients, especially phosphorus, in bottom
deposits.
Recommendation: The Subcommittee wishes to stress
that the concentrations set forth are suggested solely as
guidelines and the maintenance of these may or may
not prevent undesirable blooms All the factors causing
nuisance plant growths and the level of each which
should not be exceeded are not known
(1) In order to limit nuisance growths, the addition
of all organic wastes such as sewage, food processing,
cannery, and industrial wastes containing nutrients,
vitamins, trace elements, and growth stimulants should
be carefully controlled. Furthermore, it should be
pointed out that the addition of sulfates or manganese
oxide to a lake should be limited if iron is present in
the hypolimmon as they may increase the quantity of
available phosphorus
(2) Nothing should be added that causes an in-
creased zone of anaerobic decomposition of a lake or
reservoir
(3) The naturally occurring ratios and amounts of
nitrogen (particularly NOa and NHi) to total phos-
phorus should not be radically changed by the addition
of materials As a guideline, the concentration of total
phosphorus should not be increased to levels exceeding
100 ng/\ in flowing streams or 50 ^g/l where streams
enter lakes or reservoirs
(4) Because of our present limited knowledge of
conditions promoting nuisance growth, we must have a
biological monitoring program to determine the effec-
tiveness of the control measure put into operation A
monitoring program can detect in their early stages the
development of undesirable changes in amounts and
kinds of rooted aquatics and the condition of algal
growths With periodic monitoring such undesirable
trends can be delected and corrected by more stringent
regulation of added organics
Toxic substances
Aquatic life too frequently is considered only
in terms of harvestable species. The fact that nu-
merous other organisms are essential to produce
a crop of fishes often is overlooked or given little
attention. To produce a harvestable crop of fish,
it is essential to have supporting plants and ani-
mals for food Requirements are established on
the basis that the needed criteria are those that
will protect fish, the harvested crop, and the food
organisms necessary to support that crop. At this
time, it is believed that every important species
should be protected. One can appreciate that un-
important organisms may be sacrificed if the
following criteria are adopted. Fish too often
are considered as a single species instead of a
multitude of species. Many are distinctly and
greatly different from other related species and
have their own distinctive requirements. Because
of this, and because the important species, essen-
tial food organisms, and water quality will be
different in different habitats, a single value or
concentration has very limited applicability unless
appropriate margins of safety are incorporated.
For these reasons, the bioassay approach de-
scribed later in this section is favored It is believed
that bioassays are the best method for determining
safe concentrations of toxicants for the species
of local importance Bioassays are essential also
to determine safe concentrations for food orga-
nisms of those species and the effect of existing
water quality, including environmental variables
as well as existing pollution. Pertinent to this
stance is the fact that the majority of specific pol-
lution problems are ones involving discharges of
unknown and variable composition. Almost with-
out exception, more than one toxicant or stress is
present Further, suggested safe concentrations
probably will not be adequate in instances where
more than one adverse factor exists. It is believed
that these recommended levels will be adequate
for a particular pollutant if dissolved oxygen,
temperature, and pH are within the limits recom-
mended If the latter parameters are outside
recommended limits, appropriate alterations in the
criteria for toxicants must be made.
Most of the available toxicity data are reported
as the median tolerance limit (TLm), the concen-
tration that kills 50 percent of the test organisms
within a specified time span, usually in 96 hours
or less This system of reporting has been mis-
applied by some who have erroneously inferred
that a TL„, value is a safe value, whereas it is
merely the level at which half the test organisms
are killed In many cases, the differences are great
between TL„, concentrations and concentrations
that are low enough to permit reproduction and
growth
Substantial data on long-term effects and safe
levels are available for only a few toxicants, per-
haps 10 The effect of toxicants on reproduction
is nearly unknown, yet this is a very important
aspect of all long-term toxicity tests. In chronic
tests with six different toxicants, there were three
-------
toxicants with which certain concentrations per-
mitted indefinite survival and normal appearance
but blocked spawning completely. Such evidence
makes estimates of safe concentrations based on
acute lethal test data alone very difficult and fre-
quently erroneous. Equally problematical is the
near-total lack of information on the sensitivity
of the various life stages of organisms. Many or-
ganisms are the most sensitive in the larval,
nymph, molting, or fry state; some may be the
most sensitive in the egg and sperm stage
A further difficulty is encountered in recom-
mending criteria because continuous acceptable
concentrations must be lower than the intermittent
concentrations that may be reached occasionally
without causing damage. There seems to be only
one way in which to resolve this difficulty and
that is to use both maximum concentration and a
range of concentrations. It is recognized that the
extremes do limit organisms, but, within these ex-
tremes there is a range in concentration that can
be tolerated and is safe for prolonged periods of
exposure
Average 24-hour concentrations can be deter-
mined by using a small water pump to collect 1 to
5 ml samples every minute. After 24 hours, the
sample is mixed and analyzed. The concentration
found represents the average concentration. Sam-
ples obtained this way are more reproducible and
easier to secure than the maximum instantaneous
concentration. Maximum concentrations must be
considered in the criteria, however, because an
average concentration alone could be met and yet
permit a lethal concentration to exist for a critical
period.
Bioassay
The use of some type of bioassay to determine
the toxicity of a material or waste can be the most
effective and accurate method of assessing poten-
tial danger With these methods, no assumptions
need be made concerning the chemical structure
or form of the pollutant, nor does the investigator
have to know the constituent substances The ef-
fects of water quality on toxicity also may be
measured Naturally, the more that is known about
the chemical and physical behavior of a toxicant
in water, the more precise the assay can be.
While there are many types of assays, two
are in general use: (1) the static bioassay in which
the test solution is not changed during the period
of exposure, and (2) the flow-through bioassay
in which the test solution continually is renewed.
It is nearly impossible in a static test to use the
introduced test concentration for calculating TLm
values, especially for substances or wastes that are
toxic at concentrations of 1 mg/1 or less, because
the quantity taken into the test organism may be
a very large percentage of the amount contained
in the test water. A 48-hour TLm based on the
introduced concentration could give a value much
higher than the true concentration because of this
decrease in toxicant concentration The initial test
concentration is usually not measured in static
tests because of the changing concentration.
Knowledge of the concentration of the toxicant
at the end of the test can be of value.
The staUc test can give useful relative measures
of toxicity for wastes of high toxicity, but for the
reason mentioned above, it should not be used
for absolute values. Less toxic substances can be
assayed much more accurately and lethal concen-
trations can be determined with confidence. The
chemical nature of the tested substances has an
important effect on the accuracy of the results
as well. Substances that are volatile, unstable, or
relatively insoluble may not be accurately assayed
while substances having opposite properties can
be assayed more accurately.
The problem of maintaining oxygen concen-
trations suitable for aquatic life in the test chamber
can be very difficult Insufficient oxygen may be
present in the test water volume because a BOD
or COD may consume much of the available dis-
solved oxygen and aeration or oxygenation may
degrade or remove the test material. Devices for
maintaining satisfactory dissolved oxygen in static
tests have been proposed and used with some de-
gree of effectiveness A rather complete account of
static assays can be found in Sandard Methods
for the Examination of Water and Wastewater,
12th edition (1965), and Doudoroff, et al.
(1951)
In the flow-through type of bioassay a device
is used to add toxicant to a flow of water and the
mixture is discharged into the test container. This
method of testing has few of the problems men-
tioned in connection with the static test and has
other advantages in addition. Its important dis-
advantage is the more complicated work of build-
ing the necessary equipment; namely, a water
supply system, metering devices, and the provision
of a large quantity of the test substance.
Its important advantages are that a predeter-
mined concentration of test matenal can be main-
tained, oxygen concentrations can be kept high
or be controlled, metabolites and waste products
are removed (animals can be fed), absolute rather
than relative TLm values can be obtained, and
volatile, unstable, and sparingly soluble materials
can be tested. Additionally, multifactor experi-
ments are possible in which several variables can
be controlled (pH, dissolved oxygen, carbon diox-
-------
ide, etc.). The constant renewal test is superior for
monitoring effluents, water supplies, or streams on
a continuous or intermittent basis and is the only
suitable method for long-term tests.
Several systems for adding the test materia! to
the water have been devised since this type of
bioassay has been in use. Lemke and Mount
(1963) describe a system using a controlled water
flow balanced against a chemical metering pump
Henderson and Pickering (1963) describe a sim-
ple drip system and a controlled water flow, a
similar system is proposed by Jackson and Brungs
(1966) Both of these latter references describe
the use of fish and flowing systems as continuous
monitors. Mount and Warner (1965), Mount and
Brungs (1967), and Brungs and Mount (1967)
describe systems suitable for continuous short or
long-term tests
Most criteria for toxic substances must be based
on a bioassay made for each specific situation
This is dictated by the lack of information and
the wide variation in situations, species, water
quality, and the nature of the substance being
added to the water.
Most of the bioassay work on algae has meas-
ured the threshold concentrations that reduce phys-
iological processes by 50 percent rather than the
concentrations that cause 50-percent death in the
population tested. It is very difficult to determine
the death point of algae cells, but some workers
have used it as a criterion Physiological measure-
ments have been based largely on 50-percent
reduction in photosynthesis and 50-percent reduc-
tion in number of divisions that have taken place
during a period of time. This is determined by
the number of cells present at the beginning and
end of the experiment A bioassay method employ-
ing diatoms has been recognized by the American
Society for Testing Materials (1964)
Application Factors
Short-term or acute toxicity tests provide in-
formation on the overall toxicity of a material
and thus precede meaningful long-term toxicity
studies They may also be used to compare toxic-
ities of different materials When water for dilu-
tion is taken from the receiving stream, these tests
may also indicate additional stresses due to mate-
rials already present in the receiving water These
acute studies do not indicate concentrations of a
potential toxicant that are harmless under condi-
tions of long-term exposure It is desirable, there-
fore, to develop a factor that can be used with 96
or 48-hour TLm values to indicate concentrations
of the waste or material in question that are safe
in the receiving water Such a factor has been
called an application factor
Ideally, an application factor should be deter-
mined for each waste or material. To do this, it
would be necessary to determine the concentration
of the waste or material in question that does not
adversely affect the productivity of the aquatic
biota on continuous exposure, in water of known
quality, and under environmental conditions (DO,
temperature, pH, etc ) at which it is most toxic
This concentration is then divided by the 96-hour
TLn, value obtained under the same conditions
to give the application factor.
safe concentration for continuous exposure
96-hour TLm
For example, if the 96-hour TLra is 0 5 mg/1 and
the concentration of the waste found to be safe
is 0 01 mg/l, the application factor would be:
safe concentration _0 01 _ 1
96-hour TLm - 0 50 ~~ 50
In this instance, the application factor is Vf>o or
0 02. Then in a given situation involving this
waste, the safe concentration in the receiving
stream would be found by multiplying the 96-hour
TL„, by 0 02.
To effectively determine the application factor
for a given waste, it is necessary to determine the
concentration of that waste which is safe under
a given set of conditions For those materials
whose toxicity is not significantly influenced by
water quality and in streams free of other wastes
that influence the waste in question or that have
water qualities similar to those under which the
waste was tested, the above-mentioned concen-
tration would be the one that is safe in the re-
ceiving water However, differences in water qual-
ity and lack of information on the toxicity of
waste materials already present make the direct
use of laboratory-determined safe levels unwise
at present, and a different approach is recom-
mended.
In this approach, a 96-hour TL,„ is determined
for the waste using water from the receiving stream
for dilution and, as test organisms, the most sensi-
tive species or life stage of an economically im-
portant local species or one whose relative sensi-
tivity is known. This procedure would take into
consideration the effects of local quality and the
stress or adverse effects of wastes already present
in the stream. The TLm value thus found then is
multiplied by the application factor for that waste
to determine the safe concentration of that waste
in that stream or stream section. Such bioassays
should be repeated at least monthly and at each
change in process or rate of waste discharge.
This procedure must be used because of the
-------
extreme difference in sensitivity among species and
among necessary fish food organisms. Henderson
(1957) has discussed various factors involved
in developing application factors. Results of studies
by Mount and Stephan (1967), in which con-
tinuous exposure was used, reveal that the appli-
cation factor necessary to reduce the concentration
low enough to permit spawning ranged from 1/!
to Y:,qo- It is recognized that exposure will not be
constant in most cases and that higher concentra-
tions usually can be tolerated for short periods.
At present, safe levels have been determined
lor only a few wastes and hence only a few appli-
cation factors are known. Since the determination
of safe levels is an involved process, it will be nec-
essary to use indirect or stopgap procedures for
estimating tolerable concentrations of various
wastes in receiving waters To meet this situation,
it is proposed to use three universal application
I actors selected on the basis of present knowl-
edge, experience, and judgment. It is proposed
that these general application factors be applied
lo TLm values determined by those discharging
wastes in the manner described above to set toler-
able concentrations of their wastes in the receiv-
ing stream.
It should be evident that when these general
application factors are used for all wastes the re-
sulting concentrations at times will be more strin-
gent than needed for some wastes and inadequate
lor others The derived concentrations will be
lolerable, however, for a considerable number of
wastes in the midrange of relative toxicity.
Recommendation for the Use of Bioassays and Appli-
cation Factors To Denote Safe Concentrations of
Wastes in Receiving Streams: (1) For the deter-
mination of acute toxicities, flow-through bioassays are
the first choice. Methods for carrying out these flow-
through tests have been described by Surber and
Thatcher (1963), Lemke and Mount (1963), Hender-
son and Pickering (1963), Jackson and Brungs (1966),
Mount and Warner (1965), Mount and Brungs (1967),
and Brungs and Mount (1967) Flow-through bio-
assays should be used for unstable, volatile, or highly
toxic wastes and those having an oxygen demand They
also must be used when several variables such as pH,
DO, COi and other factors must be controlled
(2) When flow-through tests are not feasible, tests
of a different type or duration must be used The kinds
of local conditions affecting the procedure might be a
single application of pesticides or lack of materials and
equipment
(3) Acute static bioassays with fish for the deter-
mination of TLm values should be carried out in ac-
cordance with Standard Methods for the Examination
of Water and Wastewater, 12th edition (1965). Such
tests should be used for the determination of TLm val-
ues only for persistent, nonvolatile, or highly soluble
materials of low toxicity which do not have an oxygen
demand because it is necessary to consider the amount
added as the concentration to which the test organisms
are exposed
(4) When application factors are used with TLm
values to determine safe concentrations of a waste in
a receiving water, the bioassay studies to determine
TLm values should be made with the most sensitive
local species and life stages of economical or ecological
importance and with dilution water taken from the
receiving stream above the waste outfall Other species
whose relative sensitivity is known can be used in the
absence of knowledge concerning the most sensitive of
the important local species or life stages or due to
difficulty in providing them in sufficient numbers
Alternatively, tests may be earned out using one species
of diatom, one species of an invertebrate, and two
species of fish, one of which should be a pan or game
fish Further, these bioassays must be performed with
environmental conditions at levels at which the waste is
most toxic Tests should be repeated with one of the
species at least monthly and when there are changes in
the character or volume of the waste
(5) Concentration of materials that are nonper-
sistent (that is, have a half life of less than 96 hours)
or have noncumulative effects after mixing with the
receiving waters should not exceed Ho of the 96-hour
TLm value at any time or place. The 24-hour average
of the concentration of these materials should not
exceed V£o of the TLm value after mixing. For other
toxicants the concentrations should not exceed and
Hoo of the TLm value under the conditions described
above. Where specific application factors have been
determined, they will be used in all instances.
When two or more toxic materials whose effects are
additive are present at the same time in the receiving
water, some reduction in the permissible concentrations
as derived from bioassays on individual substances or
wastes is necessary The amount of reduction required
is a function of both the number of toxic materials
present and their concentrations in respect to the de-
rived permissible concentration An appropriate means
of assunng that the combined amounts of the several
substances do not exceed a permissible concentration
for the mixture is through the use of following rela-
tionship
(£+£ +£si)
Where C., Cb, C„ are the measured concentra-
tions of the several toxic materials in the water and
L., Lb, L„ are the respective permissible concen-
tration limits denved for the matenals on an individual
basis Should the sum of the several fractions exceed
one, then a local restriction on the concentration of
one or more of the substances is necessary
Heavy Metals
An extensive discussion of the physiological
mode of action of heavy metals is found in the
toxicity portion of the section on water quality re-
quirements for marine organisms.
Zinc: While much information has been pub-
lished regarding zinc, a large amount of the data
cannot be used because of incomplete description
-------
of methods, type of water, or concentrations. The
authors of many of the papers dealing with zinc
toxicity have used various specific sublethal effects
as endpomts and there is no way to compare these
findings with other work
Since the concentration of calcium and mag-
nesium influences heavy metal toxicity, permissible
levels of heavy metals are dependent on the cal-
cium and magnesium concentrations. Certain stud-
ies with zinc (Mount, 1966; and unpublished work
of the FWPCA National Water Quality Lab., Du-
luth, Minn.) and cadmium indicate that for a
given calcium and magnesium concentration the
acute toxicity of zinc and cadmium increases
(TLra concentration decreases) as pH is raised
from 5 to 9. This seems contrary to prevalent
opinion that metal toxicity is related to metal in
solution and that as pH increases (solubility de-
creases) the toxicity decreases. The reason for this
apparent contradiction is that conceptions con-
cerning the effect of pH are based on natural
waters in which pH does not vary independently
of calcium and magnesium concentrations, but
rather is closely related to it. In those cases where
this relationship has been studied, except for one
(Sprague, 1964b), the toxicity has increased with
an increase in pH. This concept also is consistent
with the work of Lloyd (1961b) and, more re-
cently, that of Herbert and Wakeford (1964) who
concluded that colloidal or flocculated, but sus-
pended, zinc exerts a toxic influence on fish.
The significance of temperature and the cal-
cium-magnesium content on the toxicity of zinc
to plankton has been pointed out by Patrick (un-
published data). In these tests, a 50-percent re-
duction in growth of the population was used as
a measure. Results of these tests are summarized
as follows:
Concentration in mg/i which reducei
growth of population by SO percent
Ca-Ug concentra- Ca-Mg concentra-
tion—44 mg/l as lion—170 mg/l as
Temperature ol CaCOi Nllzchia CaCOt Navlcula
test solution linearis serainolum
72 F 4 29 mg/l 4 05 mg/l
82 F. _.l 59 mg/l 2 31 mg/l
86 F __J 32 mg/L— 3.22 mg/l
Palmer (1957) found that zinc dimethyl dithio-
carbamate (ZDD) inhibited growth of Micro-
cystis at 0.004 mg/l A concentration of 0.25 mg/l
controlled all diatoms, 43 percent of the blue-
green algae, and 18 percent of the green algae.
The above evidence implies that permissible levels
of zinc cannot be related to the calcium-magne-
sium concentrations or to pH alone
Herbert and Wakeford (1964) described the
effect of salinity on the toxicity of zinc to. rain-
bow trout Since zinc was most toxic to trout in
freshwater, it is assumed that concentrations which
are safe in freshwater will be safe for the salmonids
in brackish water. The maximum reported affect
of a reduction of dissolved oxygen from 6-7 mg/l
to 2 mg/l on the acute toxicity of zinc is a 50-
percent increase in its acute toxicity (Lloyd,
1961a, Pickering, in press; Cairns and Scheier,
1958a). Since 4 mg/l is the minimum permitted,
this effect is small in comparison to the differ-
ence between safe and acutely toxic concentra-
tions. The use of an application factor, there-
fore, should provide adequate protection. Simi-
larly, Herbert and Shurban (1963a) found that
the 24-hour TLm for zinc was reduced only 20
percent for rainbow trout forced to swim at 85
percent of their maximum sustained swimming
speed
The effect of calcium and magnesium con-
centrations on the toxicity of zinc for plankton,
invertebrates, fishes, and their embryonic stages
is reflected in the spread of values reported as
toxic by many sources (Anderson 1950, Brungs,
in press; Cairns and Scheier 1957, 1958b; Grande,
1966, Herbert and Shurben, 1963a, b, Jones,
1938, Lloyd, 1961b, Patrick, personal communi-
cation; Pickering, in press, Pickering and Hender-
son, 1966a, Pickering and Vigor, 1965, Skid-
more, 1964, Sprague, 1964a, b, Sprague and
Ramsey, 1965; Williams and Mount, 1965; and
Wurtz, 1962)
Recommendation: The relationship between calcium
and magnesium concentration, pH, and zinc toxicity is
confusing and the separate effects have been little
studied Brungs (in press) has determined that Hoo of
the 96-hour TLm value is a safe concentration for con-
tinuous exposure
Copper: The same general considerations apply
to the determination of safe levels of copper as
apply to safe levels for zinc and the discussion of
copper will be based on the same assumptions.
From the published data, differences in species
sensitivity to copper appear to be somewhat greater
than for zinc (Anderson, 1950; Grande, 1966;
Herbert and Vandyke, 1964; Jones, 1938; Lloyd,
1961b, Mount, in press', Pickering and Hender-
son, 1966a; Sprague, 1964a, b; Sprague and
Ramsey, 1965, Trama, 1954; Turnbull, DeMann,
and Weston, 1954). Mount (in press) has found
that V& o of the 96-hour TLm value is a safe concen-
tration for continuous exposure of fish.
Bringmann and Kuhn (1959a, b) report that
0 15 mg/l copper is the threshold concentration
which produces a noticeable effect on Scenedes-
mus. Maloney and Palmer (1956) report that 0.5
-------
mg/1 copper as copper sulfate produces the fol-
lowing percents of death in algae:
57 percent in 17 species of blue-green algae
35 percent in 17 species of green algae
100 percent in 6 species of diatoms
Fitzgerald, Gerloff, and Borg (1952) report that
0 2 mg/1 of copper (as copper sulfate) produces
a 100-percent kill of Microcystis aeruginosa.
Crance (1963) found 0.05 mg/1 kills Microcystis.
Hassall (1962), working with Chlorella vulgaris
found that 25 g/1 of copper sulfate did not in-
hibit respiration if cultures were shaken. If shaking
stopped, concentrations greater than 250 mg/1
were toxic. Preliminary experiments indicate that
lack of air increases toxicity of copper Hale
(1937)—according to Jordan, Day, and Hendrix-
son (1962)—reported that the following concen-
trations were necessary to control the indicated
algae:
0 5 mg/1—Cladophora
0 1 mg/1—Hydrodictyon
0 12 mg/1—Sptrogyra
0 20 mg/1—Ulothrix
Calcium and magnesium concentrations are usu-
ally not given for algal tests, but it would seem
that the concentrations deemed safe for fish
would also be acceptable for plankton.
Recommendation: The maximum copper (expressed
as Cu) concentration (not including copper attached
to silt particles or in stable organic combination) at
any time or place should not be greater than V$0 the
96-hour TLm value, nor should any 24-hour average
concentration exceed Vi0 of the 96-hour TLm value
Cadmium: Few studies have been made of the
toxicity of cadmium in the aquatic environment.
Mammalian studies have shown it to have sub-
stantial cumulative effects. Permissible levels in
drinking water are 0 01 mg/1 (USDHEW,
1962b), and concentrations of a few p.g/g in
food (McKee and Wolf, 1963) have caused
sickness in human beings. Mount (1967) found
accumulations in living bluegills as high as 100
/ig/g (dry weight) and in the gills of dead catfish
up to 1000 /ig/g. Little accumulation was found
in the muscle. Consideration should be given to
acceptable residue levels in fish when establishing
cadmium criteria.
Daphma appears to be very sensitive to cadmium
(Anderson, 1950). Bringmann and Kuhn (1959a)
indicate Scenedesmus, and Escherichia coli are
about equally sensitive. Data as yet unpublished
(Pickering, in press) reveal that following pro-
longed exposure there is a large accumulation of
cadmium in fish. Even though very little data are
available yet, the evidence warrants a more re-
strictive requirement for cadmium than specified
under the general bioassay section.
Recommendation: The concentration of cadmium
must not exceed '^o of the 96-hour TL„ concentration
at any time or place and the maximum 24-hour average
concentration should not exceed V&oo of the 96-hour
TLm concentration.
Hexavalent Chromium: The chronic toxicity
of hexavalent chromium to fish has been studied
by Olson (1958) and Olson and Foster (1956,
1957). Their data demonstrate a pronounced
cumulative toxicity of chromium to trout and sal-
mon Mr. P. A. Olson (personal communication)
of Battelle Memorial Institute advises that some
recent comparisons of 48 and 96-hour TLm con-
centrations with concentrations not adversely af-
fecting the same species indicate that the applica-
tion factor for hexavalent chromium is 15Aoo ooo
for salmon and %oo.ooo f°r rainbow trout. He
also feels however, that such factors are not valid
for carp. Doudoroff and Katz (1953) found that
bluegills tolerated a 45 mg/1 level for 20 days in
hard water. Cairns (1956), using chromic oxide
(Cr03), found that a concentration of 104 mg/1
was toxic in 6 to 84 hours Daphnia and Micro-
regma exhibit threshold effects at hexavalent
chromium levels of 0 016 to 0 7 mg/1.
Some data are available concerning the toxicity
of chromium to algae. The concentrations of chro-
mium that inhibit growth (Hervey, 1949) for the
test organisms are as follows. Chlorococcales, 3.2
to 6.4 mg/1, Euglenoids, 0 32 to 1 6 mg/1; and
diatoms, 0 032 to 0 32, mg/1 Chromium at sub-
lethal doses sometimes stimulates algae. Patrick
(unpublished data) has studied the effects of tem-
perature on the toxicity of chromium to certain
algae. Her findings on the concentrations which re-
duce population growth by 50 percent are as fol-
lows.
Nitzschia linearis—50 percent reduction in
growth of populaUon as compared with control
(soft water 44 mg/1 Ca-Mg as CaC03)
22 C—0 208 mg/1 Cr
28 C—0.261 mg/1 Cr
30 C—0.272 mg/1 Cr
Navicula seminulum var. hustedtii (hard water
170 mg/1 Ca-Mg as CaC03)
22 C—0.254 mg/1 Cr
28 C—O 343 mg/1 Cr
30 C—0.433 mg/1 Cr
Recommendation: Data are too incomplete to do more
than urge caution in the discharge of chromium Con-
centrations of 0.02 mg/1 in soft water have been found
safe for salmonid fishes.
-------
TABLE III-5A. Pesticides 0
INSECTICIDES
[48-hour TLu values from static bioassay, in micrograms per liter. Exceptions are noted ]
Pesticide
Stream Invertebrate1
Species TLra
Clsdocerans *
Species TLri
FIsh»
Species
Qammarus
lacustrls,*
TLra Ttm
Abate Pteronarcys 100
californica.
Aldrins _ P. californica 8
Allethrin —P. californica 28
Azodrin
Aramite
Baygon8 -P. californica—
Baytex6 —P. californica
110
130
D magna ...
D magna ...
D pulex
D. magna ...
D pulex
D pulex
D pulex
D. pulex
D pulex
D magna
D magna
Benzene hexachlonde p. californica 8
(lindane).
Bidnn P. californica 1,900
Carbaryl (sevin) P. californica 13
Carbophenothion
(tnthion).
Chlordane5 P. californica 55
Chlorobenzilate
Chlorthion
Coumaphos
Cryolite
Cyclethrm ~ II..I. IIII__
DDD (TDE)' p. californica 1,100
DDT* p californica 19
Delnav (dioxathion) ...
Delmeton (systex) . IIIIIIHIIHIIII HHH
Diazinon * . p californica 60
Dibrom (naled) p californica 16
Dieldrin p californica 1.3
Dilan
Dimethoate p cahfornica 140
(cygon).
Dimethrin
Dichlorvos ¦ (DDVP) ...pVcalifctfmca""!: ""To
Disulfoten (di-syston)..P. ca|lfornica _ 18
Dursban Peteronareella 1 8
badia
Endosulfan (thiodan) ..p californica 5 6
Endrin 5 p californica 0 8
EPH
Ethion p cahfornica 14
Ethyl guthion "
Fenthion p californica 39
Guthion® p californica 8
Heptachlor5 ...p. badia 4
Kelthane (dicofel) p. californica 3,000
Kepone
Malathion* p badi3 6
Methoxychlor1 P. californica 8
Methyl parathion5
Morestan p californica 40
Ovex P cahfornica 1,500
Paradichlorobenzene
Parathion' __P californica...
Perthane
Phosdrin ' P. californica...
Phosphamidon P californica...
Pyrethrins P californica...
Rotenone P californica...
Strobane0 P. californica...
Tetradifon (tedion)
TEPP'
Thanite
Thimet
Toxaphene4 P. californica 7
Trichlorofon P. badia 22
(dipterex).5
Zectran P californica 16 D pulex
Brook trout..
1,500
640
Daphma
28
Rainbow
3
12,000
pulex
trout.
D. pulex
21
do
19
20
do
7,000
D magna
345
Bluegill
35
100
Fathead
25
50
Simocephalus
3.1
Brown t
80
70
serrulatus.
D. pulex
460
Rainbow t ...
18
88
D. pulex
600
do
8,000
790
D pulex
64
Brown t
1,500
22
D. magna
0 009
Bluegill
225
28
S serrulatus—
20
Rainbow t —
10
80
S. serrulatus--
550
do
710
4 5
1
5,000
55
32
0 36
14
09
35
240
21
2,500
D pulex 0 07
D magna
D pulex ..
D. magna
D magna
D pulex _.
D pulex __
D magna
D. pulex ..
D. magna
240
20
01
0 01
.....
02
42
390
O pulex ..
D pulex ..
D magna
1.8
08
48
11
D pulex
04
D magna
94
9
0 pulex
. 0 16
460
D. magna
4
64
D pulex
25
900
D. pulex
10
7
D magna 450
D pulex ..
D magna
15
81
10
0.14
Rainbow t
47,000
Rainbow t —
9
18
Bass
2 1
2 1
Bluegill
14
690
do
81
do
30
500
Brook t.
78
160
Bluegill
34
1,000
do
16
600
do
9,600
400
Rainbow t
700
Bluegill
700
1
do
40
70
Rainbow t -.-
20
0.4
do
1 2
64
Bluegill
02
47
do
17
36
do
230
32
Rainbow t ...
Rainbow t...
10
0.3
do
9
100
do
100
do
37.5
Brook t
19.5
1.8
Rainbow t ...
7 2
1 3
Bluegill
8,000
do
96
do
700
Rainbow t
880
Bluegill
47
6
Rainbow t
7
do
17
310
do
8,000
38
do
54
18
Bluegill
22
350
Rainbow t ...
25
Bluegill
1,100
140
Fathead
390
52
Bluegill
55
70
Rainbow t ...
28
70
do
160
60
do
8,000
76
* See notes following Table Ill-SB
-------
Pesticides
A general description of the use and the effects
of pesticides on aquatic life is given in the marine
section. Basically, their effects are similar in both
the marine and fresh water environments
The addition of any persistent chlorinated
hydrocarbon pesticides is likely to result in dam-
age to aquatic life Therefore, as concentrations
of these chemicals increase in the aquatic environ-
ment, progressive damage will result The acute
effects usually will be recognized, but the chronic
consequences may not be observed for some time.
The use of other kinds of chemical pesticides in
or around fresh waters may produce a variety of
acute and chronic effects on fish and the other
components of the biota. Because these other
chemicals are usually not as persistent as the
chlorinated hydrocarbons, the Subcommittee feels
some of them can be used around water, but only
in amounts below those that produce chronic dam-
age to desirable species.
Recommendation: (1) Chlorinated hydrocarbons —
Since any addition of persistent chlorinated hydro-
carbon insecticides in likely to result in permanent
damage to aquatic populations, their use should be
avoided
(2) Other chemical pesticides—Addition of other
kinds of chemicals used as insecticides, herbicides,
fungicides, defoliants, acaracides, algicides, etc, can
result in damage to some organisms Table I1I-5 lists
the 48-hour TLm values for a number of pesticides for
various types of fresh water organisms To provide
reasonably safe concentrations of these materials in
receiving waters, application factors ranging from Ho
to Moo should be used with these values depending on
the characteristic of the pesticide in question and used
as specified earlier in the section on application factors
Concentrations thus derived tentatively may be con-
sidered safe under the environmental conditions rec-
ommended
Other Toxic Substances
Detergents and Surfactants: The toxicity of
ABS has been reported by many workers A wide
range of endpoints have been used as criteria and
while comparison is difficult, a reasonable conclu-
sion is possible There is no agreement on the
effect of calcium and magnesium concentration.
Recommendations are based on the data from
table III—6.
Recommendation: With continuous exposure, the con-
centration of ABS should not exceed V7 of the 48-
hour TLm concentration Concentrations as high as
1 mg/1 may be tolerated infrequently for periods not
exceeding 24 hours. ABS may increase the toxicity
of other materials
Much less work has been done on LAS, a
newer, degradable detergent, than on ABS. Bar-
dach, Fujiya, and Holl (1965) report that 10 mg/1
is lethal to bullheads and 0.5 mg/1 will erode 50
percent of the taste buds within 24 days. For fat-
head minnow fry, Pickering (1966) reports a
9-day TLni of 2 3 mg/1. Thatcher and Santner
(1967) report 96-hour TL,„ values from 3.3 to
6.4 mg/1 for five fish species. Swisher, O'Rourke,
and Tomlinson (1954), testing bluegills, found
TLm values of 3 mg/1 for LAS and 12 carbon
homologs and 0.6 mg/1 for 14 other carbon homo-
logs An intermediate degradation product had a
TLm of 75 mg/1. Dugan (1967) found that sensi-
tivity to chlorinated pesticides possibly increased
after exposure to detergent Other studies as yet
unpublished indicate a surprising increase in tox-
icity at low dissolved oxygen concentrations.
Pickering and Thatcher (in press), in the only
reported study on reproduction, found that 0 6
mg/1 had no measurable effect on reproduction or
growth but 1 1 mg/1 had an effect. In tests with
five species of fishes Thatcher and Santner
(1967) found two species which were more sensi-
tive to LAS than fathead minnows
With both ABS and LAS detergents, the more
readily degradable components are the more toxic.
As a result, the components remaining will be less
toxic than the original product.
Recommendation: The concentration of LAS should
not exceed 0 2 mg/1 of of the 48-hour TL„, con-
centration, whichever is the lower
Cyanide: Although it has been studied exten-
sively, cyanide is not well understood as a hazard
to aquatic life Certain unique and peculiar char-
acteristics necessitate special treatment of this
chemical even though acceptable concentrations
cannot be given
Recent work on fish by Doudoroff et al. (1966),
has demonstrated that HCN rather than CN is the
toxic component. Except for certain extremely
toxic heavy metals (silver, for example) the tox-
icity of metallo-cyanide complexes can also be
attributed to the HCN. This then makes the effect
of pH on cyanide toxicity of great importance.
Doudoroff (1956) demonstrated a thousandfold
increase m the toxicity of a nickelo-cyanide com-
plex associated with a drop in pH from 8 0 to 6 5.
A change in pH from 7 8 to 7.5 increases the tox-
icity ten times The data reported by Cairns and
Scheier (1963b) indicate that the calcium-mag-
nesium concentrations (hardness) do not mate-
rially affect cyanide toxicity It should be noted
that in their test solutions while the calcium-
magnesium concentration of their soft and hard
-------
TABLE III-5B. Pesticides, cont.
HERBICIDES. FUNGICIDES, DEFOLIANTS, ALGICIDES
Pesticide
Stream invertebrate 1
Species TLm
Cladocerans 3
Species TLm
Flsh»
Species
Gamma rus
lacustrls,*
TLm TLm
Ametryne -
Aminotriazole . ..
Aquathol
Atrazine
Azide, potassium
Azide, sodium
Copper chloride
Copper sulfate
Dichlobenil Pteronarcys 44,000
califorriica.
2,4-D, PGBEE
2,4-D, BEE P californica 1,800
2,4 D, isopropyl
2,4-D, butyl ester _. ..
2,4-D, butyl +
isopropyl ester
2,4,5-T isooctyl ester
2,4,5-T isopropyl ester.. __ . .. __
2,4,5-T PGBE
2(2,4-DP) BEE -
Dalapon p californica
Very low toxicity
Dead X P californica 5,000
DEF P californica 2,300
Dexon P californica 42,000
Dicamba
Dichlone
Difolitan P californica 150
Dmitrocresol P californica 560
Diquat ..
Diuron P californica 2,800
Du-ter
Dyrene
Endothal, copper
Endothal,
dimethylamme
Fenac, acid P californica 70,000
Fenac. sodium P californica 80,000
Hydram (molinate) P californica 3,500
Hydrothol 191
Lanstan (korax)
LFN ...
Paraquat p californica
Very low toxicity
Propazine
Si I vex, PGBEE
Silvex, isoctyl
Silvex, BEE
Simazme P californica 50,000
Sodium arsemte P californica
Very low toxicity
Tordon (picloram)
Tnfuraltn P californica 4,200
Vernam 1 (vernolate)
Daphma 3,600
magna
Daphma 3,700
pulex
D pulex 3,200
D magna 6,000
D pulex 3,700
D magna 26
D pulex 1,400
D magna 490
D pulex 4,500
D pulex 3,700
D. pulex 2,000
Simocephalus 1,400
serrulatus
D pulex 240
Rainbow t.
3,400
Bluegill 257
Rainbow t... 12,600
Bluegill
do ...
do ...
do ...
do ...
Rainbow t..
Bluegill _.
do
do
do
10,000
9,000
1,500
1,800
760
do
do
do
do
Rainbow t...
Bluegill
Bluegill
non tox
Rainbow t
Channel Cat-
Rainbow t...
do
do
Bluegill
1,400
980
1,100
150
20,000
960
2,100
800
1,300
1,500
16,700
1,700
560
1,100
Very Low toxicity
Rainbow t ...
9,400
36
23,000
~~~48
31
210
12,300
4,300
33
15
290
5,600
230
6,000
5,800
11,500
6,500
380
do
. 1,150
do
. 16,500
do
. 7,500
18,000
do
290
do
690
1,000
do
100
5,500
do
79
Very low
18,000
toxicity
Rainbow t...
. 7,800
do
650
Bluegill
. 1,400
do —
. 1,200
Rainbow t...
. 5,000
21,000
do
. 36,500
do
. 2,500
48,000
do
11
5,600
do .
. 5,900
25,000
1 Stonefly bioassay was done at Denver. Colo , and at Salt
Lake City Utah Denver tests were in soft water (35 mg/l TDS).
non-aerated, 60 F Salt Lake City tests were in hard water (150
mg/l TDS), aerated, 48-50 F Response was death
' Daphnla pulex and Simocephalus serrulatus bioassay was
done at Denver, Colo , in soft water (35 mg/l TDS), non aerated,
60 F Daphnla magna bioassay was done at Pennsylvania State
University in hard water (146 mg/l TDS), non-aerated, 68 F
Response was immobilization
5 Fish bioassay was done at Denver, Colo . and at Rome, N Y
Denver tests were with 2 inch (ish in soft water (35 mg/l TDS),
non aerated, Irout at 55 F, other species at 65 F Rome tests
were with 2-2'/i inch fish in soft water (6 mg/l TA pH 5 85-
6 4), 60 F Response was death
' Gammarus bioassay was done at Denver, Colo, in soft
water (35 mg/l TDS), non-aerated, 60 F Response was death
5 Becomes bound to soil when used according to directions,
but highly toxic (reflected in numbers) when added directly to
water
-------
water was greatly different, the pH and toxicity
were similar.
Burdick and Lipschuetz (1950) show that some
metallo-cyanide complexes decompose in sunlight
and become highly toxic due to release of cyanide
from the complex. Cairns and Scheier (1963b)
found some increase in toxicity at reduced oxygen
concentrations and Henderson, et al. (1961)
demonstrated marked cumulative toxicity of an
organic cyanide in 30-day tests.
The toxicity of cyanide to diatoms varied little
with change of temperature and was a little more
TABLE III—6. Effect of Alkyl-Aryl Sulfonate, Including ABS, on Aquatic Organisms
Organisms
Concentration (mg/l)
Time
Effect
References
Trout . 5 0
37
50
Bluegills 4.2
37
0 86
16 0
56
17 0
Fathead minnows 2.3
13 0
U 3
Fathead minnow fry 3.1
Pumpkinseed sunfish 9 8
Salmon 5 6
Yellow bullheads 1.0
Emerald shiner 7 4
Bluntnose minnow 7 7
Stoneroller 8 9
Silver jaw 9.2
Rosefin 9 5
Common shiner 17 0
Carp 18.0
Black bullhead 22 0
"Fish" 6.5
Trout sperm 10 0
Daphma 5 0
20 0
7 5
Lirceus fontinalis 10.0
Crangonyx setodactylus 1 ... 10 0
Stenonema ares .. 8.0
16 0
Stenonema heterotarsale ... 8 0
16 0
Isonychia bicolor 8 0
Hydropsychidae (mostly 16 0
cheumatopsyche)
32 0
Orconectes rusticus 16 0
32 0
Goniobasis livescens 16 0
32 0
Snail 18.0
24 0
Chlorella 3.6
Nitzchia linearis 5.8
Navicula seminulum 23 0
26 to 30 hours
24 hours
24 hours
48 hours
30 days
90 days
96 hours
96 hours
96 hours
7 days
3 months
3 days
10 days
96 hours
96 hours
96 hours
96 hours
96 hours
96 hours
96 hours
96 hours
96 hours
24 hours
96 hours
14 days
14 days
10 days
10 days
10 days
10 days
9 days
12 days
12 days
9 days
9 days
12 days
12 days
96 hours
96 hours
Death __ Wurtz-Arlet, 1960.
TLm
Gill pathology Schmid and Mann, 1961
TLm Turnbull, et al , 1954.
TLm
Safe
TLm Lemke and Mount, 1963.
Gill damage Cairns and Scheier, 1963
TLm
Reduced spawning ...Pickering, 1966
TLm Henderson, et al, 1959.
TL„, Thatcher, 1966
TLni Pickering, 1966.
Gill damage Cairns and Scheier, 1964.
Mortality Holland, et al., 1960.
Histopathology Bardach, et al , 1965.
TLm ...Thatcher, 1966.
TLm Thatcher, 1966.
TL„ Thatcher, 1966
TLm ..Thatcher, 1966.
TLm Thatcher, 1966.
TLm ..Thatcher, 1966.
TLm Thatcher, 1966.
TLm Thatcher, 1966
Min lethality Leclerc and Devlaminck,
1952
Damage Mann and Schmid, 1961.
TLm ..Sierp and Thiele, 1954.
TLm Godzch, 1961.
TLm Godzch, 1961.
6 7 percent survival... Surber and Thatcher, 1963.
(hard water).
0 percent survival Surber and Thatcher, 1963.
(hard water).
20-33 percent Surber and Thatcher, 1963.
survival
0 percent survival Surber and Thatcher, 1963.
40 percent survival...Surber and Thatcher, 1963.
0 percent survival Surber and Thatcher, 1963.
0 percent survival Surber and Thatcher, 1963.
37-43 percent Surber and Thatcher, 1963.
survival
20 percent survival...Surber and Thatcher, 1963.
100 percent survival..Surber and Thatcher, 1963.
0 percent survival Surber and Thatcher, 1963.
40-80 percent Surber and Thatcher, 1963
survival
0 percent survival Surber and Thatcher, 1963.
TL,„ -Cairns and Scheier, 1964.
TLm .Cairns and Scheier, 1964.
Slight growth Maloney, 1966.
reduction
50 percent reduc- Cairns, et al., 1964.
tion in growth
in soft water.
50 percent reduc- Cairns, et al., 1964.
tion in growth
in soft water.
1 Misidentified originally as Synurella
-------
toxtc in soft water than in hard water (Patrick,
unpublished data). For Nitzchia linearis a 50-per-
cent reduction in growth of the population in soft
water (44 mg/1 Ca-Mg as CaC03) occurred as
follows: 0 288 mg/1 (CN) at 72 F, 0.295 mg/1 at
82 F, and 0.277 mg/t at 86 F For Navicula semi-
nulum var. hustedtii, the concentrations that re-
duced growth of the population 50 percent in hard
water (170 mg/1 Ca-Mg as CaC03) were as fol-
lows 0 356 mg/1 at 72 F, 0 491 mg/1 at 82 F,
and 0 424 mg/1 at 86 F.
Recommendation: Permissible concentrations of cy-
anide should be determined by the flow-through bio-
assay method described in the bioassay section These
tests should be conducted with DO, temperature, and
pH at recommended levels for the factors under which
the cyanide (HCN) is most toxic or under local water
conditions at which it is the most toxic.
Ammonia: The toxicity of ammonia has been
studied by several investigators but because of in-
adequate reporting and unsatisfactory experi-
mental control, much of the work is not usable
Doudoroff and Katz (1950), Wuhrmann, et al.
(1947), and Wuhrmann and Woker (1948) give
a complete account of the pH effect on ammonia
toxicity and demonstrate that toxicity is dependent
primarily on undissociated NH4OH and nonionic
ammonia. They found no obvious relationship be-
tween time until loss of equilibrium and total
ammonium content. They also demonstrated a
striking synergy between ammonia and cyanide.
McKee and Wolf (1963) state that toxicity is in-
creased markedly by reduced dissolved oxygen.
Field studies by Ellis (1940) and other observa-
tions lead to the conclusion that at pH levels of 8 0
and above total ammonia expressed as N should
not exceed 1 5 mg/1 It has been found that
2.5 mg/1 total ammonia expressed as N is acutely
toxic.
Recommendation: Permissible concentrations of am-
monia should be determined by the flow-through bio-
assay with the pH of the test solution maintained at
8 5, DO concentrations between 4 and J mg/1, and
temperatures near the upper allowable levels
marine
and estuarine
organisms
Others: Especially significant sources of wastes
that must be considered individually are derived
from tar, gas, and coke-producing plants, pulp
and paper mills, petroleum refining and petro-
chemical plants, waterfront boating activities, and
special-purpose laboratories. These problems are
discussed in the toxicity portion of the section on
water quality requirements for marine organisms.
-------
ESTUARIES are recognized as being of critical
importance m man's harvest of economically
useful living marine resources It is in these areas
that the maximum conversion of solar energy into
aquatic plant life takes place and they are justly
identified as "nurseries" since so many animals
utilize them for feeding their early life stages. Some
species, such as the oyster, spend their entire life
span in the estuary, while the shrimp and men-
haden reside there only as juveniles. The salmon
and a few others use the estuary primarily as a
pathway In sum, however, more than half of the
over 4 5 billion pounds of fishery products har-
vested by U.S. fisherman annually is derived from
animals dependent for their existence on clean
estuarine waters during some part or all of their
life cycle.
Pollution of estuarine and coastal waters is diffi-
cult to assess because of the special qualities of
this environment Technically, any foreign sub-
stance or environmental condition that inter-
feres with a desired use may be considered a pol-
lutant, but we are concerned with those substances
present at high enough concentrations or en-
vironmental changes great enough to cause de-
leterious effect Many naturally occurring sub-
stances in salt water become toxic when their
concentrations are increased artificially or by
natural processes
The problem in establishing criteria in estuaries
arises from the fluctuating nature of the water
quality, both daily and seasonally, and geograph-
ically within the estuary Changes in salinity, pH,
turbidity, and temperature may alter greatly the
critical toxic concentration of a pollutant Most
chlorinated hydrocarbon pesticides, for example,
are significantly more acutely toxic at summer
rather than winter'water temperatures and at least
one of the common detergents becomes decidedly
more toxic to fish as salinity levels increase
The most obvious effect of tidal action in the
estuary is to change water depth This indirectly
changes current patterns, water temperature, and
the density of motile animal populations Depend-
ing on the geography of the estuary and the
amount of fresh water drainage into it, salinity
patterns may vary from relatively uniform condi-
tions throughout a tidal cycle to situations in
which the> water is clearly stratified with a layer of
relatively fresh water overlying the bottom salt
water, or to situations m which the major portion
of the water mass changes from fresh to salt and
back to fresh again
In shallow, broad estuaries, wind may be the
dominant factor in causing water movements
which change salinity and temperature patterns.
The volume of fresh water discharged into an
estuary may be a major factor in establishing
coastal currents that transport pollution loads from
one region to another.
We are dealing, then, with an environment in
which the characteristics of the receiving water are
usually fluctuating, frequently unexpectedly. As a
result, its ability to dilute and disperse a burden of
toxicants is unpredictable without detailed local
investigations
Pollution in the estuary may be derived from
contamination hundreds of miles upstream in the
river basin or it may be of purely local origin Silt
plays a major role in the transport of toxicants,
especially pesticides, down to the estuary. Agri-
cultural chemicals are adsorbed on silt particles
Under poor farming practices, as much as 11 tons
of silt per acre per year may be washed by surface
water into a drainage basin. Surface mining and
deforestation further accelerate the process of ero-
sion and permit the transport of terrestrial chemi-
cal deposits to the marine environment
Atmospheric drift is also an important factor in
the transport of pollutants to the aquatic environ-
ment (Cohen and Pinkerton, 1966) Much of the
tonnage of aerially applied pesticides fails to reach
the designated spray areas and the presence of
5 /ig/l of DDT in presumably untreated Alaskan
rivers indicates the magnitude of this facet of the
pollution problem The continuous presence of
5 /xg/l of DDT in the marine environment would
decrease the growth of oyster populations by
nearly 50 percent
Toxic pollutants may be passed directly into
the marine environment as contaminants of in-
dustrial and domestic waste effluents or they may
be intentionally placed there as in the control of
various noxious insects by spraying marsh and
littoral habitats with synthetic pesticides. Experi-
mentally, some synthetic insecticides have been
applied directly to estuarine bottoms in efforts to
control oyster pests.
Finally, there are naturally occurring substances
such as lignins and phosphate compounds which
in times of flood may be carried to the estuary in
sufficient quantity to constitute a pollution hazard.
Salinity
The spatial and temporal distribution of salinity
profoundly affects the activities of many estuarine
species in tidal tributaries (Andrews, 1964; Emery
and Stevenson, 1957, Hargis, 1965 and 1966;
Pearse and Gunter, 1957, Pntchard, 1953). Some
bottom organisms, e g, Crassostrea virginica, are
-------
able to survive lower salinities than can their
predators and disease-causing organisms. Hence,
in some tidal tributaries, oysters thrive in regions
where they are sheltered from these pests by low
salinity. Natural alterations in salinity distribution
have been reportedly followed by increased mor-
tality of oysters. It is clear that care must be
exercised in the approval of engineering projects
or industrial processes that will alter salinity re-
gimes in tidal tributaries and lagoons and in their
associated wetlands
Salinity patterns can be caused to vary from
"normal" by alterations in character of freshwater
inflow and basin geometry. These are the same
factors that produce changes in circulation In fact,
salinity alterations are precursors to changes in
density currents
Recommendation: For the protection of estuarine
organisms, no changes in channels, in the basin geom-
etry of the area, or in fresh water inflow should be
made that would cause permanent changes in tsohaline
patterns of more than ±10 percent of the natural
variation
Currents
Despite their large volumes, tidal waters, espe-
cially those in tributaries of the seas, have special
circulatory characteristics that may affect their
ability to assimilate wastes For example, tidal ac-
tion slows the already slowed (due to lowered
slope and resulting reduced speed of gravity-in-
duced flow) seaward movement of water in tidal
rivers and streams This alternate up and down
stream movement of the water in the freshwater
portion of the tidal tributary is confounding
enough in itself (Ketchum, 1950 and 1951; Stom-
mel, 1953a, b) but in the estuarine reach, the area
where sea salts are noticeable, further complexi-
ties often occur (Bowden, 1963, Hargis, 1965;
Redfield, 1951) In horizontally and vertically
stratified mixing estuaries, there are two streams
The upper stream, fresher and lighter, has a net-
flow downstream while the lower stream, saltier
and heavier, flows inward or upstream Since
these surface currents and bottom counter-cur-
rents often extend far to sea off the mouths of
large tidal tributary or estuarine systems, as well as
far upstream, significant upstream transport of ma-
terials in solution or suspension in the counter-
current can occur These circulatory features are
important in the life cycles of many estuarine
species. For example, oyster and barnacle setting is
related to tidal and nontidal currents (Barlow,
1955; Bousfield 1955; Emery and Stevenson,
1957; Hargis, 1966; Ketchum, 1954, Pritchard,
1953). Large disturbances of current patterns can
disrupt the life cycles of estuarine organisms.
Hence, projects that alter current patterns should
be carefully evaluated and controlled.
It is possible to alter circulatory patterns in tidal
tributaries by (1) changing the quantity, timing,
and location of fresh water inflow, (2) changing
the geometry of the basin. The former can be ac-
complished by construction and operation of reser-
voirs above or below the fall line (defined as the
uppermost limit of ocean's tidal activity). The
latter can be accomplished by shoreline or bottom
modification; e g, drainage, bulkheadmg and fill-
ing, channel dredging, and subaqueous spoil dis-
posal or mining. Oyster harvesting practices have
been known to produce marked changes in bot-
tom geometry (Hargis, 1966)
Recommendation: In view of the requirements of
estuarine organisms and the nature of marine waters,
no changes in basin geometry or fresh water inflow
should be made in tidal tributaries which will alter
current patterns in such a way as to cause adverse
effects
PH
Despite the great emphasis given to the impor-
tance of pH in the literature, little is known of its
direct physiological effects on marine organisms.
Its indirect effects, however, are extremely sig-
nificant Even a slight change in pH indicates that
the buffering system inherent in sea water has been
altered radically and that either a potential or
actual carbon dioxide imbalance exists. This im-
balance can be deleterious or disastrous to marine
life A second indirect effect is that pH can in-
fluence the toxicity of other materials Cyanide and
ammonia, discussed under "Toxicity," are out-
standing examples of this kind of action.
Recommendation: Materials that extend normal ranges
of pH at any location by more than ±0 1 pH unit
should not be introduced into salt water portions of
tidal tributaries or coastal waters At no time should
the introduction of foreign materials cause the pH to
be less than 6 7 or greater than 8 5
Temperature
Temperature requirements of marine and estua-
rine organisms in the biota of a given region may
vary widely Therefore, if we are to maintain tem-
perature favorable to the biota, all important spe-
cies, including the most sensitive, must be pro-
tected. It has been found that organisms in the
intertidal zone vary considerably in their ability
to withstand high temperatures Those in the up-
permost areas of the tidal zone generally can with-
-------
stand higher temperatures than those in the lower
portions of the tidal zone and these in turn gen-
erally can withstand higher temperatures than the
same species of animals living in the subtidal
zones. In addition, when considering the coastline
as a whole, we must recognize that there ate vari-
ous races within a given species which may vary
considerably in their environmental requirements,
or in their ability to withstand extreme conditions.
In our marine waters, there is a great mixture
of species Species typical of higher latitudes are
found with species that are more abundant farther
south Tropical or subtropical species generally
will spawn in the summer months. Species from the
higher latitudes require low water temperature for
spawning and the development of the young Thus,
they usually spawn in the winter months and tem-
peratures at that time are critical Any warming of
the water during the cold weather or winter pe-
riod could be disastrous from the standpoint of the
elimination of the more northerly species. In some
instances, a rise in winter temperatures of only 2
or 3 F might be sufficient to prevent spawning and
thus eliminate these species from the biota
In the northern portions of the country there is
generally a great range in natural temperatures In
southern areas, as we approach the tropics, we find
smaller overall temperature ranges. In the tropics
or subtropics, optimum temperatures for many
forms are only a few degrees lower than maximum
lethal temperatures Great care should be exer-
cised, therefore, to prevent harmful increases in
maximum summer temperatures in tropical areas
In general, temperatures in the marine waters
do not change as rapidly nor do they have the
overall range from extreme to extreme as they do
in fresh waters. Marine and estuanne fishes, there-
fore, are less tolerant of temperature variation.
They can accommodate somewhat, but overall
temperature range and rate of change are even
more important here than they are in fresh waters.
It has been observed that when surface water tem-
peratures over the Georges Bank increased from
46 to 68 F, the larval fish died at 65 F. It has been
found that species in subtropical and tropical en-
vironments are living at temperatures that are only
a few degrees less than their lethal temperatures
In the most northern forms, extensive variations
in seasonal temperatures are a necessity for orderly
development and growth. Spawning and develop-
ment frequently occur at lower temperatures and
the sexual products ripen on rising temperatures
after a period of low temperatures Temperatures
above or below the optimum range may delay or
speed up development They may inhibit swim-
ming ability and the effectiveness of food utiliza-
tion may be decreased with increasing tempera-
tures in the upper viable range. Fishes and other
forms are also more susceptible to parasites and
diseases at temperatures outside of their optimum
range. In regard to rapid changes in temperature,
it has been found that a drop in temperature from
58 to 43 F kills sardines. Tolerable temperature
minima vary with the population and its past tem-
perature history. Kills have occurred off the Texas
coast at 40 F whereas kills of the same species
have occurred off Bermuda at a drop to 59 F.
Many kills have occurred in nature due to unusu-
ally low temperatures. Kills also occur due to
natural high temperatures. Yellowtail flounder and
whiting larvae died when they drifted from an area
of 44 F to one of 64 F It has been reported that
61 F is best for the developing of mackerel, but
70 F is too high These are merely illustrations of
what might happen to species occurring in inshore
waters
It is apparent from the foregoing that data are
very sparse on temperature requirements of marine
and estuarine species It is very difficult, therefore,
to attempt to suggest temperature requirements
for marine and estuanne forms The difficulty is
compounded by the great extent of the Nation's
shorelines, the differing natural temperature varia-
tions from north to south, and the geographic over-
lapping of species native to different latitudes
Consideration must be given to maximum allow-
able temperatures for both the summer period and
the winter period.
In attempting to establish permissible levels of
temperature increase in receiving waters due to
heated waste discharges, precaution must be taken
to prevent—
(a) excessive incremental increases above
background values even though such in-
cremental increases lie below maximum
limits, and
(b) exceeding maximum natural background
limits
Such precautions are necessary to prevent grad-
ual net increases in background temperatures due
to the continuously increasing volumes of heated
wastes being discharged into receiving waters
The discharge of heated wastes into estuaries
and other tidal tributaries must be managed so that
no barrier to the movement or migration of fish
and other aquatic life is created
Recommendation: In view of the requirements for
the well-being and production of marine organisms, it
is concluded lhal Ihe discharge of any heated waste
into any coastal or estuarine waters should be closely
managed Monthly means of the maximum daily tem-
peratures recorded at the site in question and before
-------
the addition of any heat of artificial origin should not
be raised by more than 4 F during the fall, winter, and
spring (September through May), or by more than
1 5 F during the summer (June through August)
North of Long Island and in the waters of the Pacific
Northwest (north of California), summer limits apply
July through September, and fall, winter, and spring
limits apply October through June The rate of tem-
perature change should not exceed 1 F per hour except
when due to natural phenomena
Suggested temperatures are to prevail outside of
established mixing zones as discussed in the section on
zones of passage
Dissolved oxygen
Dissolved oxygen requirements of marine orga-
nisms are not as well known as those for freshwater
forms Studies have been made indicating that
minimum dissolved oxygen concentrations of 0 75
to 2 5 are required for test species to resist death
for 24 hours. Most marine species died when the
dissolved oxygen dropped below 1.25 mg/1 for a
few hours. Reduced swimming speed and changes
m blood and serum constituents occurred at dis-
solved oxygen levels of 2.5 to 3 mg/1 It was
found that DO levels between 5 3 and 8 mg/1 were
satisfactory for survival and growth Levels above
17 mg/1, however, produced adverse effects Large
fluctuations in dissolved oxygen from 3 to 8 mg/1,
diurnal or otherwise, produced significantly more
physiological stress in fishes than fluctuations from
3 to 6 mg/1 In tests made to date, it has been
found that 5 to 8 mg/1 of DO is apparently suffi-
cient for all species of fish for good growth and
general well being It is generally recognized that
in deeper waters DO values are often considerably
less than 5 0 mg/1 In estuaries where there is a
reduction in salinity, levels may drop to as low as
4 mg/1 at infrequent intervals and for limited pe-
riods of time It is probable that many marine ani-
mals can live for long periods of time at much
lower levels of DO. Experimental studies with
freshwater organisms have demonstrated, how-
ever, that low concentrations of DO at which adult
fishes can live almost indefinitely, can inhibit feed-
ing and growth In determining DO requirements,
it is essential to consider growth, reproduction,
and other necessary life activities.
Recommendation: For the protection of marine re-
sources, it is essential that oxygen levels shall be suffi-
cient for survival, growth, reproduction, the general
well-being, and the production of a suitable crop To
attain this objective, it is recommended that dissolved
oxygen concentrations in coastal waters, estuaries, and
tidal tributaries of the Nation, including Puerto Rico,
Alaska and Hawaii, should be as follows.
(I) Surface dissolved oxygen concentrations in
coastal waters shall not be less than 5.0 mg/1, except
when natur.il phenomena cause this value to be de-
pressed.
(2) Dissolved oxygen concentrations in estuaries
and tidal tributaries shall not be less than 4 0 mg/1
at any time or place except in dystrophic waters or
where natural conditions cause this value to be de-
pressed
The committee would like to stress the fact that, due
to a lack of fundamental information on the DO re-
quirements of marine and estuarine organisms, these
requirements are tentative and should be changed when
additional data indicate that they are inadequate
Crude oil and petroleum products
The discharge of crude oil and petroleum prod-
ucts into estuarine and coastal waters presents spe-
cial problems in water pollution abatement. Oils
from different sources have highly diverse proper-
ties and chemistry Oils are relatively insoluble in
sea and brackish waters and surface action spreads
the oil in thin surface films of variable thickness,
depending on the amount of oil present. Oil, when
adsorbed on clay and other particles suspended in
the water, forms large, heavy aggregates that sink
to the bottom. Additional complications arise
from the formation of emulsions in water, leach-
ing of water soluble fractions, and coating and
tainting of sedentary animals, rocks, and tidal
flats
Principal sources of oil pollution are numerous.
Listed in order of their destructiveness to ecosys-
tems, they are.
(1) Sudden and uncontrolled discharge from
wells towards the end of drilling operation
(2) Escape from wrecked and submerged oil
tankers.
(3) Spillage of oil during loading and unload-
ing operations, leaky barges, and accidents
during transport
(4) Discharge of oil-contaminated ballast and
bilge water into coastal areas and on the
high seas.
(5) Cleaning and flushing of oil tanks at sea.
On the average, a ship's content of such
wastes is estimated to contain 2 to 3 per-
cent oil in 1,000 to 2,000 tons of waste.
(6) Spillage from various shore installations,
refineries, railroads, city dumps, garages,
and various industrial plants.
Spillage From Wrecked Oil Tankers
Even though wrecks of oil tankers along the
Atlantic coast and subsequent spillage of oil into
the sea have been reported several times, no thor-
-------
ough examination has been made of the effect of
oil pollution on local marine life, except for fre-
quent references to the destruction of waterfowl
One of these disasters attracted the general atten-
tion of the public and members of the Audubon
Society of New England One night in 1952, two
tankers, the Fort Mercer and the Pendleton, went
aground on the shoal of Monomoy Point, Cape
Cod. Large amounts of oil spilled from the broken
vessels, spread long distances along the shore, and
were responsible for high mortality of ducks (scot-
ers and eiders). Many thousands of oil-smeared
dying birds were seen along the coast Attempts to
save some of the birds by removing the oil with
various solvents failed No published records are
found on the effect of this massive spillage on
aquatic life. According to the records of the Mas-
sachusetts Audubon Society, serious oil spreads
threatening fish and bird life have occurred at least
six times since 1923 along the beaches of Cape
Cod The latest occurrence was on Sunday,
April 16, 1967. Heavy films of crude oil appeared
along the coast from Chatham to Provincetown,
Mass. and spread to Cape Cod Bay, Nantucket
Island, and Boston. The shores of the National
Seashore Park were seriously affected and hun-
dreds of ducks and brant were found dead or
dying.
The massive spillage of oil may constitute a
disaster of a national and even international mag-
nitude as has been dramatically demonstrated by
the wreck in March 1967 of the super tanker Tor-
rey Canyon carrying 118,000 tons of crude oil
About one-half of the load gradually spilled near
Seven Stones Reef, off the southern coast of Eng-
land, where the tanker was stranded By the middle
of April, patches of crude oil began to appear on
the French coast in Brittany, threatening the pro-
ductive oyster farms in the inlets and estuaries It
is obvious that a disaster of such magnitude is be-
yond the scope of an ordinary pollution problem
in coastal waters The probability of a recurrence
of heavy oil spillage is, however, very real because
of the present trend in the methods of transporting
oil in very large and apparently vulnerable tankers
It has been reported that Japan operates a tanker,
Idemitsu Maru, of 205,000 Ions holding capacity.
A super tanker of over 300,000 tons capacity is
under consideration and a design of a 500,000-
ton tanker appeared in the press
Effect of Oil Spillage on Aquatic Life
of a Small Marine Cove
W. J, North, et al (1965) made a valuable
study of the effect of massive spillage of crude oil
into a small cove in lower California Bay Prior to
the spillage, the investigators were engaged in a
study of bottom fauna and flora of the cove and
were in possession of background information
which made it possible for them to record the
changes that took place after the water of the cove
was contaminated by the 59,000 barrels of oil that
escaped from the wreck of the tanker Tampico on
March 29, 1957. Among the many dead and dying
species observed a few weeks after the disaster, the
most frequently found were abalones {Haliotts
julgens, H rujescens, and especially H. crachero-
du), lobsters (Panulirus interruptus), pismo clams
(Tivela stultorum), mussels (Mytilus sp.), sea
urchins (Strongylocentrotus franciscanus, S pur-
puratus), and sea stars (Pisaster giganteus, P
ochraceus) A slight improvement of the bottom
fauna was noticeable a few months after the dis-
aster, but extensive recovery became apparent only
2 years later. Four years after the accident, the
populations of abalones and sea urchins still were
reduced greatly and seven species of animals pre-
viously recorded in the cove had not been found
at all.
Combined Effect of Oil and Sewage Pollution
The oil and sewage pollution effects on aquatic
organisms of the Novorossiyak Bay (Black Sea,
U.S.S.R ) was recently studied by Kalugtna, et al
(1967) For a number of years, this bay has been
receiving a mixed daily discharge of 15,000 to
30,000 cubic meters of petroleum refinery wastes
and domestic sewage. There is marked decrease of
various valuable species of mollusks (Spisula
subtruncata, Tapes rugatus, Pecten ponticus) and
complete destruction of oyster beds (Ostrea
taurica) due to the combined effect of pollution
and depradations by a carnivorous gastropod
(Rapana) Samples were collected 1 to 25 meters
from the outfall for bioassay Copepods (Acartia
clausi) placed in samples taken 25 meters from
the outfall were killed in 24 hours Larvae of
decapods and gastropods in samples taken 10 to
25 meters out perished in 3 to 4 days Calanus was
killed in 5 days in samples taken 1 meter out, but
survived the 10-day test in the samples taken 5,
10, and 25 meters from the outfall. There also was
a noticeable change in the distribution and species
composition of benthic algae.
Color of Oil Film on the Surface of Water
The color of the oil film on the water surface is
indicative of the thickness of the slick and may be
used as an indicator of the volume of oil spilled.
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According to data published by the American
Petroleum Institute (1949), the first trace of color
that may be observed as a surface film on the sea
is formed by 100 gallons of oil spread over 1
square mile. Films of much darker colors may
indicate 1,332 gallons of oil per square mile. Ex-
periments conducted by the Committee on the
Prevention of Pollution of the Seas (1953) showed
that 15 tons of oil covered an area of 8 square
miles In 8 days, it had drifted about 20 miles
from the point of discharge The same committee
(1953) indicated another source of oil pollution
that should not be neglected It has been found that
unburned fuel oil escaping through the funnels of
oil-burning ships may comprise 1 to 2 percent of
the total oil consumed and it may be deposited on
the sea surface. British investigators attributed the
disappearance of eel grass (Zostera) to minute
quantities of oil. Oil weakens the plant and makes
it susceptible to attacks of a parasitic protozoan
(Labyrinthula). Observations made several years
ago at Woods Hole showed that young Zostera
that began to reappear in local bays after several
years of absence were already infected by this
microorganism even though they appeared to be
healthy
Adsorption of Oil by Sand, Clay, Silt, and
Other Suspended Particles
Oil of surface films is easily adsorbed on clay
particles and other suspended materials, forming
large and relatively heavy aggregates that sink to
the bottom The surface of the water may appear
free from pollution, until the sediment is stirred by
wave action and the released oil floats up again.
During World War II, a product known as
"carbonized sand" was manufactured for the U S
Navy and used for the primary purpose of rapidly
removing oil spilled or leaked from ships Carbon-
ized sand was used principally as a rapid method
to prevent and stop fires Sand and oil aggregates,
being much heavier than sea water, sank very
rapidly and remained on the bottom Experimental
work has shown that the toxic effect of oils is not
diminished by this method (Chipman and Galts-
off, 1949). Since the end of World War II, a num-
ber of preparations to be used as solvents, emulsi-
fiers, and dispersing agents of oil slicks in harbor
waters appeared in New Zealand, Western Europe,
and the United States These preparations are be-
ing offered under various trade names and their
chemical composition is not always stated. It is
often claimed that such compounds remove oil
slicks more efficiently than mopping with straw or
coarse canvas fabric (skrim), a method exten-
sively used m Auckland Harbor (Chitty, 1948).
It is, however, generally recognized that various
detergents and emulsifiers are toxic to aquatic life
and therefore compound the danger of oil pollu-
tion. Mechanical means such as preventing the
spread of a slick by surrounding it with floating
barriers (plastic booms), spreading sawdust and
removing an oil aggregate by scooping or raking,
and erecting grass or straw barriers along the
beaches are probably more effective at present
than the chemical methods of dispersing or dis-
solving oil Even anchoring oil by combining it
with relatively heavy carbonized sand seems to be
preferable to chemical methods.
Toxicity of Crude Oil and Petroleum Products
Oil may injure aquatic life by direct contact
with the organism, by poisoning with various water
soluble substances that may be leached from oil,
or by emulsions of oil which may smear the gills or
be swallowed with water and food. A heavy oil
film on the water surface may interfere with the
exchange of gases and respiration.
A number of observations have been recorded
of the concentrations of oil in sea water which are
deleterious to various species Experimental data,
however, are scarce and consequently the toxicol-
ogy of oil to marine organisms is not well under-
stood
Nelson (1925) observed marine mollusks (Mya
arenaria) being destroyed by oil on tidal flats of
Staten Island, N.Y. The Pacific coast sea urchin,
Strongylocentrotus purpuratus, dies in about I
hour in a 0 1 percent emulsion of diesel oil. After
20 to 40 minutes in this concentration the animals
fail to cling to the bottom and may be washed
away (North, et al., 1964).
Crude oil absorbed by carbonized sand does not
lose its toxicity. This has been shown by laboratory
experiments conducted at Woods Hole (Chipman
and Galtsoff, 1949). The amount of oil used was
limited to the quantity held in the sand, hence no
free oil was present in the water. The oil-sand
aggregates were placed in containers filled with sea
water but never came into contact with the test
animals Four species were bioassayed- the very
hardy toadfish (Opsanus tan) in the yolk sac
stage, the moderately tolerant barnacle (Balanus
balanoides), and oyster (Crassostrea virginica),
and the extremely sensitive hydrozoan, (Tubularia
crocea)
The survival of toadfish embryos was indirectly
proportional to the concentration of oil in water.
-------
In a concentration of 0 5 percent, the embryos sur-
vived for 13 days (end of test); in 1.25 percent,
8i days; in 2.5 percent, 6 days; and in 5 percent,
4i days. Barnacles suffered 80 to 90-percent
mortality within 70 hours in 2.0-percent mixtures
of oil in sea water. They became inactive in 23
hours in concentrations of 2 percent and above.
Tubularia suffered 90 to 100-percent mortality
within 24 hours after being placed in water con-
taining a 1:200 oil-carbonized sand aggregate.
Water extracts of crude oil were lethal within 24
hours at concentrations of 500 mg/1 and greater.
Experiments with oysters consisted primarily of
determining the effect of oil adsorbed on carbon-
ized sand on the number of hours the oysters re-
main open and feeding and on the rate of water
transport, across the gills. A paste-like aggregate of
oil in carbonized sand (50 ml crude oil to 127 g
sand) was prepared, wiped clean of excess oil, and
placed in the mixing chamber. Sea water was de-
livered through this chamber to the recording ap-
paratus at a rate slightly in excess of the rate of
water transport by oyster gills (Galtsoff, 1964;
Chipman and Galtsoff, 1949). There was a notice-
able decrease in the number of hours the test
oysters remained open and in the daily water trans-
port rate through the gills. The time open was re-
duced from 95 to 100 percent during the first 4
days of testing to only 19 8 percent on the 14th
day. The total amount of water transported per
day, and presumably used for feeding and respira-
tion, was reduced from 207 to 310 liters during the
first 6 days to only 2 9 to 1 liter per day during
the period between the eighth and 14th day of
continuous testing These tests indicate that oil
incorporated into the sediments near oyster beds
continues to leach water-soluble substances which
depress the normal functions of the mollusk.
Critical observations are lacking on the effect of
oil on pelagic larvae of marine invertebrates, but
there is good reason to assume that crude oil and
petroleum products are highly toxic to free-swim-
ming larvae of oysters Speer (1928) considers
that they are killed by contact with surface oil film.
Laboratory experience of Galtsoff (unpublished
records) shows that oyster larvae from 5 to 6 days
old were killed when minor quantities of fuel oil
were spilled by ships in the Woods Hole harbor
and the contaminated water penetrated into the
laboratory sea water supply.
The tests described above leave no doubt that
water-soluble substances are leached from oil
spilled into water and adversely affect marine life.
It is reasonable to assume that the water soluble
materials of oil may contain various hydrocarbons,
phenols, sulfides, and other substances toxic to
aquatic life. The water-soluble fraction leached
from crude oil is easily oxidized by aeration and
loses its toxicity (Chipman and Galtsoff, 1949).
Carcinogenic Substances From
Oil-Polluted Waters
Presence of hydrocarbons similar to benzo-
pyrene in oil-polluted coastal waters and sediments
of France m the Mediterranean was reported by
Mallet (1965) and Mallet and Sardotr (1965).
The effluents from the industrial establishments on
the shores at Villefranche Bay comprise tar sub-
stances, which contain benzopyrenes, benzo-8,
9-fluoranthene, dibenzanthracenes, chrysene, 10-
methyl anthracene, and nitrogenous derivatives
such as dimethylbenzacridine. These substances
are carried out into the bay water and settle on the
bottom. The pollution is augmented by incom-
pletely burned oils discharged by turbine ships.
The content of benzopyrene in bottom sediments
ranges from 500 micrograms in 100 g sample col-
lected at the depth of 8 to 13 cm to 1 6 micro-
grams at 200 cm. Similar contamination is of im-
portance in the Gulf of Fos, Etang de Berne, and
in the delta of the Rhone River.
Carcinogenic hydrocarbons were found to be
stored in plankton of the bay of Villefranche, in
concentrations varying from 2 5 to 40 micrograms
per 100 g. Fixation of benzopyrenes was found
also in the bodies of holothurians (Lalou, 1965)
in a bay near Antibes. The reported concentration
in the visceral mass of holothunan was slightly
higher than that in the bottom sediment.
Observations on storage of carcinogenic com-
pounds found in oil-polluted water are biologically
significant. The important question of biological
magnification as these compounds are ingested by
plankton feeders remains unanswered and needs
to be investigated.
Sampling of Oil-Polluted Sea Water
The question of the minimal concentration of oil
and petroleum products consistent with unin-
hibited growth and reproduction of aquatic species
is more difficult to answer than it is in the case of
other contaminants. As has been shown above, oil
is found in water in four distinct phases: (1) sur-
face oil film, (2) emulsion in sea water, (3) ex-
tract of water soluble substances, and (4) semi-
solid aggregate of oil and sediment covering the
bottom. Obviously, no single sample could include
all four phases and the method of sampling should
-------
vary accordingly. Collection of samples of a heavy
oil slick near the origin of spillage presents no par-
ticular difficulty because an adequate quantity may
be scooped easily and placed in a proper container.
Serious difficulty arises, however, m case of an
irridescent film of oil approaching the thickness of
a monomolecular layer Garrett (1964), made a
special study of slick-forming materials naturally
occurring on sea surfaces and demonstrated their
highly complex composition. The collection of
very thin layers of surface water was made by
means of a specially constructed plastic screen
The entrapped compounds were washed off into
a large container (Garrett, 1962) He found sur-
face-acting substances in all areas where the sea
surface was altered by monomolecular films and
concluded that "a chemical potential exists
whereby such surface alterations can occur when
conditions are suitable for the adsorption and
compression of the surface-active molecules at
the air/water boundary." The oil film at the
air/water boundary may be composed of several
interacting organic compounds. This complexity
must be kept in mind in studies of oil pollution m
sea water
If a relatively thick layer of contaminated water
is needed, the sample may be scooped or sucked
from an area of sea surface enclosed by a floating
frame Interference due to wave ripples is mini-
mized in this way.
For analysis of an oil emulsion in sea water, a
sample of a desired volume may be collected by
pump or by any type of self-closing water bottle
lowered within the surf area
For obtaining water soluble substances leached
from oil sludge, sampling should be made by
pumping or by using a water sampler lowered as
close as possible to the oil-covered bottom
Samples of oil adsorbed on sediments can be
obtained by using bottom samplers designed to
take quantitative samples.
Contamination of beaches by floating tar ballast
and cleaning water discharged by ships sailing
along our coast is of such common occurrence that
at present it is almost impossible to find a public
beach free from this nuisance. Cakes of solidified
oil tar can be picked by hand from the tidal zone
of any beach along the Atlantic and Gulf coasts.
Recommendation: Until more information on the
chemistry and toxicology of oil in sea water becomes
available, the following requirements are recommended
for the protection of marine life No oil or petroleum
products should be discharged into estuanne or coastal
waters in quantities that (1) can be detected as a
visible film or sheen, or by odor, (2) cause tainttng
of fish and/or edible invertebrates, (3) form an oil-
sludge deposit on the shores or bottom of the receiving
body of water, or (4) become effective toxicants ac-
cording to the criteria recommended in the 'Toxicity"
section
Turbidity and color
Turbidity, color, and transparency are closely
interrelated phenomena in water They must be
observed simultaneously because transparency is
a function of turbidity, water color, and spectral
quality of transmitted light For practical pur-
poses, however, it is more convenient to discuss
them separately.
Turbidity
By observing the turbidity of sea water it is pos-
sible to determine the depth of the euphotic zone;
i e., the depth in which organic carbon is produced.
Various particles suspended in water reduce the
intensity of light by absorption and scattering. In
the sea, the maximum depth of growth of attached
plants vanes It is 160 m in the Mediterranean,
30 m in Puget Sound, and 10 m off Cape Cod. In
general, benthic plants will not grow at a depth at
which the light intensity is less than 0 3 percent of
its surface value (Clarke, 1954). In any environ-
ment, the rate of photosynthesis decreases with the
attenuation of light but the respiration rate remains
approximately the same Because the role of
phytoplankton in organic production is far more
important quantitatively than that of benthic
plants, an increase in the turbidity of water di-
minishes primary productivity of the ocean bio-
mass as indicated by the rate of growth of various
planktonic algae
For each species of plant, a level of light inten-
sity may be reached at which the rate of photo-
synthesis becomes equal to the rate of respiration
This level is designated as compensation intensity
and the depth at which this value is found is called
the compensation depth For marine phytoplank-
ton, it has been determined that compensation in-
tensity is about 100 ft-candles, or 1 percent of the
value of full sunlight (Clarke, 1954) In natural
waters, the compensation depth varies; e.g, in the
Gulf of Maine it was found to be 30 m while at
Woods Hole only 7 m
In many coastal waters, the principal cause of
turbidity is the discharge of silt carried out by the
principal rivers Secchi disc readings show that the
transparency of water at the mouths of large rivers
during flood stage may be reduced to a few centi-
meters. At normal river stages, the disc may be
visible at several meters below the surface. Ob-
servations from an airplane are useful in recording
-------
the distribution of brackish, silt-laden waters along
the coast. Silting of the estuaries and adjacent
coastal water should be considered as a special
case of pollution resulting from deforestation,
overgrazing and faulty agricultural practices, road
construction, and other land management abuses.
Mixed effluents from various industrial plants
and domestic sewage increase the turbidity of
receiving water. It is difficult to distinguish be-
tween the effect of the attenuation of light due to
suspended particles and the direct effect of the
particles in suspension on the growth and physi-
ology of aquatic organisms. Natural silt taken
from the bottom of the sea and kaolin affect the
development of eggs and the growth of larvae of
oysters and hard shell clams (Mercenaria merce-
naria) In a suspension of 2 g of dry silt in a liter
of sea water, only 39 percent of oyster larvae com-
pleted development In 3 g per liter there was no
development (LoosanofT, 1962). Growth of
Mercenaria clams was retarded in the concentra-
tion of 1 to 2 g/1, but appeared to be normal at
0.75 g/1. Development was completely suppressed
in the concentration of silt from 3 to 4 g/1 (Davis,
1960). Silt concentration of 0 1 g/1 caused a 57-
percent decrease in the water transport of an aduJt
oyster. In 4 g/1, the depression was 94 percent
(Loosanoff, 1962). The turbidity used in these
experiments probably is equivalent to 750 to
4,000 mg/1 of turbidity standards, although direct
comparison of figures cannot be made accurately.
The principal significance of turbidity observa-
tions in a study of pollution is the determination of
the depth of the euphotic zone as a factor affecting
primary productivity of the sea (Ryther, 1963).
Determination of the coefficient "k" defined as the
natural logarithm of the fraction of incident light
penetrating to a given depth is of great importance
in studies of organic production. In the temperate
and northern parts of the ocean, values of "k"
range between 0.10 to 0.20 and correspond to
depths of 50 to 25 m In more turbid coastal
waters, the coefficient of extinction is as high as
1 0 and a compensation depth of 5 m is com-
monly encountered. These values may be used as
a basis for comparing the characteristics of uncon-
taminated waters with those of highly turbid and
polluted waters of coastal and inshore areas A
considerable part of the turbidity of these areas is
attributable to nonliving particles
It must remembered, also, that very high tur-
bidity of sea water may be due entirely to blooms
such as are known to occur in red tide areas
(Galtsoff, 1949) or as a result of unbalanced over-
fertilization such as is induced by organic wastes
from duck farms in Great South Bay, N.Y. Tur-
bidity may be determined practically by use of a
Secchi disc Turbidity may be determined more
accurately by using the techniques described in
Standard Methods for the Examination of Water
and Wastewater, 12th edition (1965). Any tur-
bidity of less than 1 m (by Secchi disc) or in cor-
responding Jackson units should be regarded with
suspicion and the nature of suspended material as
well as the composition of plankton determined
Color
The color of sea water, expressed as dominant
wave length in millimicrons (m>i) covers the
range from violet (400 to 465 ny) to red-purple
(530 to 700 m/i) Spectrophotometry methods, as
described in Standard Methods for the Examina-
tion of Water and Wastewater, 12th edition
(1965), should be used if careful study is re-
quired, particularly for determining the exact color
of water contaminated with industrial wastes.
Monitoring the color changes of sea water yields
information on the extent of intrusion of fresh
water into the sea, the intensity and extent of silt-
ing, the location and extent of plankton blooms,
the extent and distribution of pollution from indus-
trial waste effluents, and the presence and probable
thickness of oil film.
In brackish waters, the blue hue of the open sea
is replaced by a greenish or yellowish color. Silting
areas are recognizable by brown or yellowish dis-
coloration Red-brownish color is typical of the
red tide caused by Gymnodmium and other spe-
cies of dinoflagellates. Some of these are toxic to
fishes and benthic invertebrates (Galtsoff, 1948,
1949). Mass production of forms such as the blue-
green alga Trichodesmium gives the surface of the
sea an appearance of "green meadow" as described
for the Azov Sea by Knipowich (Galtsoff, 1949).
Swarming of Phaeocystis poucheti, P. globosa, and
Rhizosolema have been reported to extend over
hundreds of square miles of the open sea causing
a distinct brownish discoloration.
Systematic studies have not been made yet to
determine the optical characteristics of discolored
sea water. It is reasonable to expect that such an
investigation would be valuable in explaining the
cause of discoloration and, in certain instances,
may indicate the presence and nature of pollution.
Light components specific for the contaminant
entering sea water may be detected by the use of
a spectrophotometer or with the recording SPOT
spectroradiometer recently developed by Alfred C.
Konrad of the Massachusetts Institute of Tech-
nology. This type of instrument is being used at
present at the Woods Hole Oceanographic Institu-
-------
tion and is proving very useful. Spectroradiometer
observations can be made either from an airplane
or from shipboard.
Recommendation: No effluent which may cause
changes in turbidity or color should be added to, or
discharged into, inshore or coastal waters unless it has
been shown that it will not be deleterious to aquatic
biota.
Settleable and floating substances
Settleable solids entering coastal waters include
various products of forest industries such as saw-
dust, bark chips, wood fibers, sewage solids,
and many industrial wastes. The old practice of
dumping sawdust into tidal rivers was discon-
tinued long ago, but its effect is still visible in
the rivers of Maine For instance, an area of the
bottom of the Damanscotta River was still cov-
ered with a loose layer of sawdust about 2 to 3 feet
deep in 1940, although operation of the lumber
mills responsible for this deposition had ceased
more than 50 years previous The Damariscotta
kitchen-midden on the banks of the river contains
a huge accumulation of river oyster shells and
some artifacts left here by the Indians who lived
there for several centuries of pre-Colonial times
The habitat was so completely changed by pollu-
tion that at present there is hardly any benthic
organism found on this formerly productive bot-
tom (Galtsoff and Chipman, unpublished report).
Decay-resisting organic matter from wood fibers
and waterlogged bark and chips constitutes, in
places, a serious handicap to aquatic life. Settle-
able materials from mining operations and gravel
and sand washing make the bottom unsuitable for
aquatic life in the affected areas of the receiving
bodies of water. Silting may be so heavy that the
sediment brought in may completely fill the bay
One can see this in the eastern branch of Mata-
gorda Bay, Tex., an area that has been completely
obliterated within the last 25 years by the Colo-
rado River.
Dredging of bays and tidal rivers for improve-
ment of navigation occasionally presents serious
problems Benthic communities in the area near
dredging operations may be destroyed or damaged
by spoil deposition, increase in water turbidity,
release of toxic substances accumulated in the mud
of the polluted areas, and by changing the pattern
of currents in the dredged area.
Careful studies of the effects of dredging on
oyster-producing bottoms of the Santee River,
S C, were made in 1936 by G. Robert Lunz, Jr
(unpublished report), for the U.S Corps of Engi-
neers No deleterious effect on oyster-producing
bottoms was found. An examination made by the
Bureau of Fisheries Laboratory at Woods Hole of
dredging operations to deepen and enlarge the
Cape Cod Canal disclosed that several productive
oyster beds near the site of dredging were covered
by 2 to 3 feet of sand and silt. The oysters were
destroyed, but the grounds soon were re-populated
by hard-shell clams and the productivity of the
area restored.
Disposal of the huge quantities of garbage ac-
cumulated by large cities presents a special and
difficult problem. The old practice of barging this
waste out to sea and dumping it is highly objection-
able Incineration seems to be the answer. This
creates, however, the problem of proper incinera-
tion of large quantities of materials without in-
creasing air pollution over the metropolis The city
of Boston disposes of large amounts of accumu-
lated garbage and trash by incineration and by
dumping the ashes into the sea at a distance from
shore State and Federal authorities are engaged
presently in a study of the chemical composition
of ash and its possible effect on aquatic life in the
sea Preliminary analysis of an incinerated sample
made by Ronald Eislcr (personal communication)
of the National Marine Quality Laboratory of the
Federal Water Pollution Control Administration
shows that aluminum, iron, and calcium were most
abundant, followed by zinc, sodium, potassium,
and lead. Other metals comprising more than 1
percent of the fraction soluble in 6nHC1 include
barium, chromium, and magnesium It is evident
that ash from this waste contains a fairly large
percentage of heavy metals which may be accumu-
lating in the bodies of fish and shellfish The effect
of ash on the behavior of fish is now being studied,
but the results are not yet available.
Examples of industrial effluents containing ma-
terials that precipitate in sea water are the waste
from titanium paint plants or the soap portion of
the effluents from Kraft pulp mills. This fraction of
the black liquor is precipitated from solution by
salt, carried by the current of the receiving river,
and eventually deposited on the bottom (Galtsoff,
et al, 1947). Waste from several plants extracting
titanium dioxide from llemenite (ferrous titanate)
produces serious pollution in the lower Patapsco
River area near Baltimore. Because of the re-
stricted circulation of water in the upper Chesa-
peake Bay, the effect is quite pronounced. Ferric
hydroxide flocculation in the Patapsco River has
been found detrimental to plankton. Diatoms were
destroyed by flocculation and removed from
plankton by settling with the iron particles. Con-
siderable amounts of iron accumulated on the
bottom and iron precipitate was found coating the
-------
gills of minnows, silverside, and white perch
(Olson, etal., 1941).
Recommendation: Water quality requirements for
specifying the permissible limits of settleable solids and
floating materials cannot be expressed quantitatively at
present Since it is known that even minor deposits may
reduce productivity and alter the benthic environment,
it is recommended that no materials containing settle-
able solids or substances that may precipitate out in
quantities that adversely affect the biota should be in-
troduced into estuanne or coastal waters It is espe-
cially urgent that areas serving as habitat or nursery
grounds for commercially important species (scallops,
lobsters, oysters, clams, crabs, shrimp, halibut, floun-
ders, demersal fish eggs and larvae, and other bottom
forms) be protected from any infringement on natural
conditions
Tainting Substances
Substances found in industrial wastes are fre-
quently responsible for objectionable or offensive
tastes, odors, and colors of fish and shellfish Even
slight amounts of oil or petroleum products in
bays and estuaries will impart an oil or kerosene
flavor to mullet, mackerel, and other fishes and
also to oysters, clams, and mussels making them
unmarketable Oysters collected in Louisiana
waters polluted by crude oil retained a distinct
flavor and odor associated with this type of pollu-
tion for several weeks after the escape of crude oil
from wells and leaky barges had been stopped
(Galtsoff, et al, 1935).
Anaerobic conditions associated with the de-
posit of sewage sludge on the bottom are accom-
panied by the production of hydrogen sulfide, a
substance that causes black discoloration of bi-
valve shells and imparts an offensive flavor and
odor to their flesh In the waters receiving black
liquor from Kraft pulp mills in the York River,
Va., the gills and mantles of oysters developed a
gray color. This condition also is found in oysters
grown in waters receiving domestic sewage (Galt-
soff, et al, 1957).
Contamination of water with copper results in
the accumulation and storage of this metal far
above its normal content in the tissues. The cop-
per content of oyster flesh from uncontaminated
waters off Cape Cod varied from 0 170 to 0.214
mg copper per oyster or from 8.21 to 13 77 mg
per 100 g dry weight. In green colored oysters
collected from adjacent areas only slightly con-
taminated with copper salts, the copper content in
the flesh ranged from 1.27 to 2 46 mg per oyster
or from 121-71 to 271 mg per 100 g dry weight
(Galtsoff and Whipple, 1931; Galtsoff, 1964).
In a current study conducted at the Northeast
Marine Health Sciences Laboratory, at Narragan-
sett, R.I., Dr. B H. Pnngle (unpublished data)
found that the average copper content of oysters
collected from unpolluted areas along the east
shore ranged from 20 to 80 mg/l, wet weight;
oysters from areas known to be polluted contained
from 124 5 to 392 0 mg/l wet weight. The copper
content of sea water ranged between 0 0038 to
0 005 mg/l in areas not known to be polluted. In
certain polluted places, concentrations as high as
0 019 mg/l were recorded.
Other metals are easily absorbed, stored, and
concentrated by oysters in great excess of their
concentration in sea water Experimentally, it has
been shown that iron and iodine can be absorbed
within a relatively short time by oysters from water
to which these metals have been added in excess
The flavor of so-called superiodized oysters pro-
duced before World War II in Arcachon, France,
was pronounced because the iodine content of flesh
was many times higher than that in untreated
oysters (Galtsoff, 1964) The color of the oysters
was not affected
Green color of the gills of the European oyster
in France and in the American oyster occasionally
found in North Carolina and Chesapeake Bay is
due to absorption of the blue-green pigment of the
diatom, Navicuta, present in large numbers on
oyster grounds. The color is not associated with
the increased copper content of flesh (Ranson,
1927).
Recommendation: To prevent the tainting of fish and
other marine organisms, substances that produce tastes
and off-flavors should not be present m concentrations
above those shown to be acceptable by means of bio
assays and taste panels Experience has shown that test
organisms should be exposed to the materials under
test for 2 weeks at selected concentrations to determine
the maximum concentration that does not produce
noticeable off-flavors as determined by organoleptic
tests (Cooking should be done by baking the material
wrapped in aluminum foil )
Plant Nutrients and Nuisance Organisms
Plant nutrients and nuisance organisms are in-
terrelated in many ways There also are many
other factors in the environment, such as tempera-
ture and salinity, that are closely correlated and,
in many instances, seem to be contributing factors
to nuisance organisms
Man, through altering the hydrography of his
environment by building dams and diverting
waterflows from their natural courses, has pro-
duced conditions in many areas that have caused
nuisance growths and brought about an imbalance
of natural conditions He also has enriched surface
waters and created imbalances in dissolved mate-
-------
rials and organisms through careless land man-
agement and by allowing the introduction of
nutrients from sewers, food processing industries,
fertilizer plants, feed lots, and farms. As a result,
natural communities of aquatic life are altered
and the functioning of these ecosystems often
is changed severely or destroyed.
To maintain natural distribution, abundance,
and interrelations of the aquatic biota, and to
control unwanted growths, it is necessary to de-
termine and maintain levels of dissolved materi-
als required for this balance This is an extremely
difficult task, however, because there are a great
many interrelated factors that contribute to the
development of excessive populations of a species.
Although a considerable amount of work has been
done on the nutrition of aquatic organisms, most
of this work has been done on a very few different
species. Very little research has been done to deter-
mine what interaction of factors causes a shift in
diversity or in the kinds of species that compose a
community. For these reasons it is impossible to
set any definite requirements. At this time the only
meaningful thing that can be done is to develop
guidelines.
Plant Nutrients: The increase of nutrients in the
sea is accelerated by deposition of material derived
from the land as sediments from the rivers, by
settling and filling caused by water movements
produced by tide or wind, and by biological activ-
ity. To date, no serious problems resulting from
abnormal enrichment of nutrients have been iden-
tified in the open sea except perhaps locally around
outfalls that extend several miles out to sea. With
the increased disposal of wastes in the sea, this
potential problem should be carefully watched.
Estuaries and tidal embayments have long been
recognized as some of our most valuable and pro-
ductive resources They are the most ephemeral of
the natural marine habitats and consequently most
easily affected by man's activities. They serve as
sinks for most of the organic and inorganic mate-
rials resulting from land erosion Because of the
lack of scouring and the nature of the sediments
that occur in some areas, anaerobic conditions
often develop in the beds of estuaries and bays
Increases in the deposition of suspended solids
intensifies this condition. An excellent discussion
of the role of sediments in an estuary is given by
Carriker (1967).
Many industries and municipalities discharge
nutrient-rich wastes into estuaries. Because of the
nature of the estuary, these are recycled and ac-
cumulated over a period of time. Because of this
recycling, effluents with low concentrations of nu-
trients may, in time, produce serious problems.
The complete flushing of the estuary often takes
many years With controlled water discharges, this
problem may become more severe.
Plant nutrients consist of many types of chemi-
cals For example, we have the chemicals com-
monly recognized as being important in plant
nutrition such as nitrate, phosphate, sulfate, car-
bonate, calcium, magnesium, sodium, and potas-
sium. There are also the so-called "trace elements"
which are equally important but are required in
small amounts such as iron, manganese, molybde-
num, cobalt, zinc, etc. More recently the impor-
tance of organic compounds in plant nutrition has
been recognized These include vitamins, such as
vitamin B12, organic forms of nitrogen, such as
urea, various amino acids, and amides, and the
simple sugars, such as glucose.
The role of dissolved organic compounds in the
nutrition of plants as well as animals appears to be
important Darnell (1967) refers to the aquatic
medium as a "vegetable soup" to indicate its rich-
ness in dissolved organic materials The work of
Ryther (1954) points out that the organic forms
of nitrogen are best utilized by the less desirable
species (Nannachloris atomus and Stichoccus sp.).
Nitzschia, a desirable diatom, often grows poorly
in their presence This is no doubt a major reason
why sewage effluents often bring about the devel-
opment of undesirable species.
If the increased nutrients in a system are well
balanced, many species will have larger popula-
tions, the predator pressure wiLI increase, and the
productivity of the whole ecosystem will increase.
If, however, the increased nutrients are of undesir-
able composition for most forms of aquatic life, or
not in the correct ratio, excessive blooms of spe-
cies with low predator pressures may develop.
Examples of these are certain blue-green algae. Of
course, environmental factors other than nutrients
are important in the development of blooms. Any
one important factor, such as temperature, light,
or water mass stability, if limiting, may prevent
blooms even though other conditions are suitable
for their development. As a result, blooms some-
times do not develop even though most of the
conditions are favorable.
Nuisance Organisms: Nuisance organisms in
the marine environment are usually defined as
those organisms which interfere with the use that
man wishes to make of a particular water. Some
examples are abnormally abundant growths of or-
ganisms that make bathing beaches unattractive,
produce unpleasant odors, foul the bottoms of
boats, spoil the esthetic appearance of water and
the coastline, clog fishing nets, interfere with the
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flow of water within intake and effluent pipes, and
interfere with navigation. This category of nuisance
organisms should also include those organisms that
interfere with the growth and reproduction of or-
ganisms important to man. For example, excessive
populations of boring sponge or oyster drills,
rooted and floating aquatics can interfere with the
movement and reproduction of fish; bacteria and
red tide organisms such as Gymnodinium and
Gonyaulax may have toxic effects on other orga-
nisms, including man (Rounsefell and Nelson,
1966, Felsing, 1966).
The groups of organisms that may cause nui-
sances or become severe pests include algae (in-
cluding red tide organisms), coelenterates,
sponges, mollusks, such as oyster drills and mus-
sels, and crustacea. These organisms are com-
monly encountered in the natural marine environ-
ment. Organisms may become nuisances because
of excessive growth and changes in distribution
patterns and predator-prey relationships. The main
causative factors are excessive and, often, imba-
lanced nutrients, considerable changes in the na-
tural regimes of temperature, turbidity, and salin-
ity, and changes in current patterns
In some instances, nuisance growths seem to be
directly related to the nutrients that are available.
In other situations, nuisance growths may not be
directly affected by artificial enrichment, so far as
we know, and seem to be more strongly affected
by changes in the temperature, salinity, or turbid-
ity. Included here are various fouling organisms,
barnacles, mussels and other mollusks, polyzoa
tube worms, marine borers, and pests to useful
marine products (oyster drills, boring sponges,
crabs, parasitic fungi, and protozoans), and
swarms of jellyfish, which make bathing in some
coastal waters hazardous during certain seasons.
The effect of increased nutrients may be an in-
crease in the populations of certain species already
present in the environment and a decrease of spe-
cies that are not tolerant of such nutrients. Exam-
ples of such conditions are the increase of Entero-
morpha and sea lettuce, Ulva laciuca, in the zone
of mineralization of sewage which occurs in some
areas of the lower Potomac. In areas of higher
salinity, abundant growths of Ascophyllum often
occur in waters containing mineralized effluents
from sewage treatment plants. In Biscayne Bay,
Fla., the following organisms became abundant
under such conditions- the flowering plants, Halo-
phila bailloms and Diplanthera wrightii, and the
echinoderm, Amphioplus abditus. Under heavy
organic enrichment, the algae, Gracilaria blodgetiii
and Agardhiella tenera, the worm, Diopatra cu-
pera, and the amphipods, Erichthonius brasihensis
and Corophtum acherusicum, became very com-
mon (McNulty, 1955).
In other cases, an unbalanced organic enrich-
ment together with changes in temperature and
salinity brings about an almost complete change in
the species composing an aquatic community plus
excessive growths of some species. An excellent
example of this type has been described by Ryther
(1954) in his studies of Moriches Bay and Great
South Bay, Long Island. In this area, duck farm
wastes enrich the bay waters with organic com-
pounds that produce a low nitrogen-to-phosphorus
ratio At the times of the largest algal blooms, low
salinities and high temperatures exist in the area.
As a result, desirable marine diatom species of
Nitzschia which prefer cool water (5 to 25 C), ni-
trates, and nitrites as a source of nitrogen, and are
not benefited by a low N/P ratio (5:1) were re-
placed by Nannochloris aiomus and Stichococcus
sp These species can grow well in nitrates, nitrites,
ammonia, urea, uric acid, and cystine, and prefer
a N/P ratio of 5.1. As Ketchum (1967) points
out, these weed species are undesirable food
sources and the natural productivity of the estuary
is destroyed Ketchum also points out that the
greatest amount of plankton does not always oc-
cur in the waters of greatest enrichment. This is
because the development of a maximum standing
crop of phytoplankton is also governed by the con-
centration of predators, stability of the water
column, transparency of the water, etc.
Nutrient imbalance may affect the ratio of inor-
ganic phosphate to total phosphorus, here defined
as the sum of inorganic, organic, and particulate
phosphorus It is known from the work of Pomeroy
(1960) and others that inorganic phosphorus is
rapidly taken up by actively growing plants At the
same time, inorganic phosphorus is regenerated as
a result of bacterial degradation and excretion by
animals. The net effect over the short run is to pro-
duce a steady state between the various fractions
of phosphorus in the environment There should
be some ratio of inorganic to total phosphorus in
the euphotic zone that would be characteristic of a
balanced nutrient regime and this ratio should be
lower than the same ratio for the imbalanced
system in which inorganic phosphorus can
accumulate. 1
Data from Moriches Bay and Great South Bay
on Long Island, Charlestown Pond, R.I, the North
Atlantic, and the North Pacific have been ex-
amined In the obviously polluted portion of Mori-
ches Bay, the inorganic total phosphate ratio gen-
erally exceeds 0 6, while the Charlestown Pond, an
uncontaminated estuary of similar characteristics
to Moriches Bay, this ratio was less than 0.4. In
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the open ocean at high latitudes and in the winter
when phytoplankton density is low, the fraction of
inorganic phosphorus may increase to 0 65 or
thereabout.
Recommendation: The ecological factors most often
associated with nuisance growths are changes in the
natural temperature and salinity cycles and increases
in nutrients The change in any of these factors may
directly or indirectly affect the response of the orga-
nisms to other factors Increase or decrease in current
and, indirectly, its effect on available nutritional ma-
terials have also been found to be important
To maintain a balance among nutrients and a bal-
anced biota most conducive to the production of a
desired crop, it is recommended that
(1) No changes should be made in the basin geom-
etry, current structure, salinity, or temperature of the
estuary without first studying the effects on aquatic life
For example, these studies should be made before dams
are erected, water diversion projects are constructed,
or dredge and fill operations carried out
(2) The artificial enrichment of the marine en-
vironment from all sources should not cause any major
quantitative or qualitative alteration in the flora Pro-
duction of persistant blooms of phytoplankton, whether
toxic or not, dense growths of attached algae or higher
aquatics or any other sort of nuisance (hat can be
directly attributed to nutrient excess or imbalance
should be avoided Because these nutrients often are
derived largely from drainage from land, special atten-
tion should be given to correct land management in a
river basin and on the shores of a bay to prevent ero-
sion
(3) The naturally occurring atomic ratio of NOrN
to POi-P in a body of water should be maintained
Similarly, the ratio of inorganic phosphorus (ortho-
phosphate) to total phosphorus (the sum of inorganic
phosphorus, dissolved organic phosphorus, and par-
ticulate phosphorus) should be maintained as it occurs
naturally Imbalances have been shown to bring about
a change in the natural diversity of the desirable orga-
nisms and to reduce productivity
Toxic Substances
Relatively few of the many substances recog-
nized as potential toxic pollutants of the marine
environment have been studied sufficiently to en-
able us to define their maximum allowable concen-
trations Specific pollutants and classes of pollu-
tants are discussed in terms of current knowledge
In some cases, data are adequate to set definite
criteria, while in others, criteria are educated
guesses at best and can serve only as temporary
guidelines
Lethal concentrations of some persistent sub-
stances as determined by acute toxicity tests are
so low that we are not justified in allowing their
deliberate introduction into the natural environ-
ment, On the other hand, a few waste products
appear to offer little threat to the marine environ-
ment because of their rapid degradation and
dispersal.
Our concern is not primarily with what polluting
substances are present, but whether or not they are
present in sufficiendy large amounts to cause dele-
terious effects on the biota and the environment.
Many naturally occurring substances, including
clean fresh water, would be toxic if discharged into
the estuarire and coastal marine environment in
sufficiently large amounts
Determination of the toxicity of known and un-
known effluents, either simple or complex mix-
tures, can best be made by determining the reac-
tions of endemic fauna exposed to them at levels
that might be expected in receiving waters. Chemi-
cal assays may determine the presence of such pol-
lutants at levels as low as nanograms per liter, but
biological systems may be affected by even smaller
amounts Many animals have the ability to accu-
mulate toxic residues of substances present in the
environment in only trace amounts until body resi-
dues are large enough to cause damage when re-
leased internally through normal metabolic proc-
esses Animals differ in their sensitivity to the same
toxicant and it is essential that toxicity data be
related, in the final analysis, to animals of eco-
nomic importance 1
A fundamental concept in attacking the pollu-
tion problem is the assumption that effluents con-
taining foreign materials are harmful and not per-
missible until laboratory tests have shown the
reverse to be true It is the obligation of the agency
producing the effluent to demonstrate that it is
harmless rather than require pollution abatement
agencies to demonstrate that the effluent is causing
damage
Specific methods are suggested here for the
determination of the toxicity of proposed effluents
While certain procedures are desirable, they are
not always reasonable and certain permissible
alternatives are also given
Basic Bioassay Test: The basic bioassay test
shall consist of a 96-hour exposure of an appro-
priate organism, in numbers adequate to assure
statistical validity, to an array of concentrations of
the substance, or mixture of substances, that will
reveal the level of pollution that will cause (1) ir-
reversible damage to 50 percent of the test orga-
nisms, and (2) the maximum concentration caus-
ing no apparent effect on the test organisms in
96 hours. Tests should be conducted, when pos-
sible, in a "flow-through" system so that the or-
ganisms arc exposed continuously to a fresh
solution of the test material appropriately diluted
with water of the same quality as that at the site
of the proposed discharge Adequate safeguards
-------
should be taken to insure that the test will be
conducted under the least favorable environmental
conditions that are allowable in the natural en-
vironment. Tests should be conducted at water
temperatures typical of the mean of maximum
daily temperatures during critical periods at the
proposed effluent discharge site
Test organisms should be selected either on the
basis of their economic importance in the area
receiving the discharge and their sensitivity or on
the basis of their importance in the food web of
economically important animals. In the event that
organisms meeting these criteria are not suitable
or available for the confined conditions of the tests,
substitute forms endemic to the area may be uti-
lized Appropriate tests must be undertaken to
demonstrate the relative sensitivity of economi-
cally important species and substitute species to the
test material so that meaningful interpretations of
the data can be made.
Application Factor: It is recognized that the
most obviously deleterious effect of toxic sub-
stances is increased mortality. More subtle changes
such as reduced growth, lowered fecundity, altered
physiology, and induced abnormal behavior pat-
terns may have more disastrous effects on the
continued existence of the species. Evaluations of
such sublethal effects generally will provide more
meaningful guidelines.
It is recognized that there should be an applica-
tion factor for each waste or material and that
these factors may vary widely for these different
wastes and materials The concept and use of ap-
plication factors is defined and discussed at length
in the toxicity portion of the section on water
quality requirements for fresh water organisms.
Due to a lack of knowledge of application factors
for specific wastes and materials, a single applica-
tion factor to be applied to all wastes is being sug-
gested at this time. This application factor may
require a lower concentration than is necessary in
some instances, particularly for those materials
that are subject to biological degradation, but it is
known that it is not restrictive enough for some
materials. Ideally, the determination of application
factors should be the result of studies for the de-
termination of safe levels of potential toxicants
under long-term or continuous exposure The ap-
plication factor is the concentration of a material
or waste that is not harmful, divided by the 96-
hour TL,n value for that material A few applica-
tion factors have been so determined at the Bureau
of Commercial Fisheries Laboratory at Gulf
Breeze, Fla. (unpublished data). In the future, as
application factors are determined for specific sub-
stances, they will replace the recommendation for
the generalized application factor for these particu-
lar materials or wastes. It is clearly understood that
as additional data become available recommenda-
tions on water quality requirements will be
changed so that they conform with the new
knowledge
Biological Magnification: Biological magnifi-
cation is an additional chronic effect of toxic
pollutants (such as heavy metals, pesticides, radio-
nuclides, bacteria, and viruses) which must be rec-
ognized and examined before clearance can be
given for the disposal of a waste product into na-
tural waters Many animals, and especially shellfish
such as the oyster, have the ability to remove from
the environment and store in their tissues sub-
stances present at nontoxic levels in the surround-
ing water, This process may continue in the oyster
or fish, for example, until the body burden of the
toxicant reaches such levels that the animal's
death would result if the pollutant were released
into the bloodstream by physiological activity. This
may occur, as in the case of chlorinated hydrocar-
bon pesticides (such as DDT and endrin) stored
in fat depots, when the animals food supply is re-
stricted and the body fat is mobilized The appear-
ance of the toxicant in the bloodstream causes the
death of the animal. Equally disastrous is the
mobilization of body fat to form sex products
which may contain sufficiently high levels of the
pollutant so that normal development of the young
is impossible.
The biological magnification and storage of
toxic residues of polluting substances and micro-
organisms may have another serious after effect.
Herbivorous and carnivorous fish at lower trophic
stages may gradually build up DDT residues of
15 to 20 mg/1 without apparent ill effect Carniv-
orous fish, mammals, and birds preying on these
contaminated fish may be killed immediately or
suffer irreparable damage because of the pesticide
residue or infectious agent.
In the final analysis, laboratory tests alone are
not sufficient to assess completely the toxic effects
of a substance These data must be interpreted in
combination with field observations Criteria es-
tablished under the artificial conditions of labora-
tory tests will probably require adjustment m
the light of later and more prolonged field
observations
Recommendation: In the absence of toxicity data other
than the 96-hour TLm, an arbitrary application factor
of Vioo of this amount shall be used as the criterion of
permissible levels
Additional chronic exposure tests will be conducted
within a reasonable period to demonstrate that the
estimated maximum safe levels as indicated by the
-------
96-hour TLm and the application factor do not, in fact,
cause decreases in productivity of the test species dur-
ing its life history
Monitoring the Marine Environment: The chief
problem in monitoring the marine habitat for pol-
lution lies in the fact that the discharge of toxic
materials may be intermittent This is not neces-
sarily true, but it means that water samples col-
lected periodically reflect only the conditions at
the time they were collected Significantly higher or
lower levels of pollution may have existed between
sample collections A second major factor for con-
sideration is that trace amounts of pollutants or
effluent mixtures toxic to the biota may not be
readily susceptible to chemical analysis For these
reasons, the analysis of resident biota for abnor-
mal changes offers a better tool for interpreting
environmental fluctuations
Mollusks are being collected for analysis at
monthly intervals in estuaries on both the Atlantic
and Pacific coasts (Butler, 1966 a, b). Analysis
of resident populations by electron capture, gas
chromatographic techniques reveal changes in
residues of 11 of the more common organochlonde
pesticides which oysters, mussels, and some spe-
cies of clams readily store. These methods are use-
ful for rapid surveys of recent pollution. By appro-
priate spacing of samples in time and location, it
has been possible to pinpoint sources of pollution.
It is suggested that a "monitoring system of this
type, appropriately expanded to include fish and
plankton, would quickly identify areas where pol-
lution problems exist Suitable analytical tech-
niques are available to make these samples equally
useful for the identification of pollution by heavy
metals and other toxic substances
Monitoring for the presence of organophosphor-
ous materials is feasible, but less specific for indi-
vidual toxic compounds This group of pesticides
exerts its toxic effect on living systems by inhibiting
the enzyme acetylcholinesterase, which is essential
to conduction in nerve fibers The nervous tissue of
fish and some invertebrates, appropriately ana-
lyzed, reveals whether the organism has been ex-
posed to organophosphorous materials within the
past 2 to 4 weeks (Holland, et al1967) Identifi-
cation of such changes can be made before toxi-
cant levels arc high enough to cause serious
mortalities.
A particularly efficient nonspecific method for
monitoring changes in the estuanne habitat is
based on the periodic collection of sedentary ani-
mals and plants which have attached themselves to
artificial cultch plates. Squares of asbestos cement
boards placed in strategic locations will be utilized
by resident biota as a habitat At 30-day or shorter
intervals these plates can be changed, the orga-
nisms enumerated, volumetncally measured or
chemically assayed, and an index of their relative
abundance obtained (Butler, 1954).
Such plates have been maintained for nearly 20
years at one laboratory in Florida (Butler, 1965),
and they supply detailed information on the rela-
tive productivity of the environment in relation to
hydrologic changes. They will be equally useful as
monitors of newly introduced pollutants in this
area The monitoring method of choice—and there
are others besides the ones suggested—will depend
on the specific environment and the animals of
particular interest No one method will be adequate
and a combination of methods should provide the
most information in the shortest time period.
Pesticides: Pesticides may be described as na-
tural and synthetic materials used to control un-
wanted or noxious animals and plants. They exert
their effect as contact or systemic poisons, as repel-
lents, or in some cases as attractants. It is conveni-
ent to classify them according to their major usage
such as fungicides, herbicides, insecticides, fumi-
gants, and rodenticides. Although data are not
available as to the total amount of pesticides used
in the United States, total production figures (in-
cluding exports) show that more than 875 million
pounds were produced in 1965 This represents an
increase of approximately 10 percent over 1964,
and more than a fivefold increase in the past two
decades In recent years, the use of herbicides has
increased relatively more rapidly than that of other
pesticides In 1964, more than 100 million acres of
the continental United States were treated with
some kind of pesticide. The trend in pesticide pro-
duction is towards the manufacture of more granu-
lar formulations This physical adsorption of the
pesticide on clay particles makes possible better
control during application and should result in less
dissipation of the chemical into atmosphere and
into nontarget areas
Despite better control of pesticide applications,
their dispersal in drainage systems and possible
eventual accumulation in estuaries makes our
coastal fisheries especially vulnerable to their toxic
effects Estuarine oyster populations, juvenile
shrimp, crab, and menhaden, for example, all
occupy the habitat where fresh and salt water mix
and where deposition of river silt with its load of
adsorbed pollutants takes place Laboratory tests
show that these economically important animals
are especially sensitive to the toxic effects of low
levels of pesticides. Oysters, for example, will exist
in the presence of DDT at levels as high as 0 1
mg/1 in the environment. But at levels 1,000 times
less (0 1 /xg/1), oyster growth or production
-------
would be only 20 percent of normal, shrimp popu-
lations would suffer a 20-percent mortality, and
menhaden would suffer a disastrous mortality.
Some insecticides are toxic enough to kill 50 per-
cent or more of shrimp populations after 48 hours
exposure to concentrations of only 30 to 50 nano-
grams per liter of the compound.
Pesticides may be classified by their chemical
affinities and a large number of economically im-
portant insecticides are chlorinated hydrocarbon
compounds. These include the well-known DDT
and aldrin-toxaphene group Typically, these are
persistent compounds, but they may be degraded
by living systems into less toxic metabolites. As
residues in soil and marine sediments, they may
persist unchanged for many years and conse-
quently present a continuing threat to animal
communities. As a general rule, the acute toxicity
of this group of pesticides increases with the level
of metabolic activity so that their presence may
cause two or three times more damage in summer
than in winter months
The organophosphorous pesticides are also pri-
marily insecticides Typically, they hydrolyze or
break down into less toxic products much more
readily than the organochloride compounds. Prac-
tically all persist for less than a year, while some
last only a few days in the environment Most of
them are degraded rather quickly in warm water
and consequently are more hazardous to aquatic
animals at winter rather than summer tempera-
tures. They exhibit a wide range of toxicity, both
more and less damaging to marine fauna than the
organochlondes They are usually preferable as
control agents because of their relatively short life.
Other major chemical categories including the
carbamates, arsenicals, and 2,4-D and 2,4,5-T
compounds are generally, but not necessarily, less
toxic to marine biota
Pesticides registered for uses which might per-
mit their dispersal into the marine environment
must be evaluated for their toxic effect on oysters,
fish, and shrimp Consequently, there is a con-
siderable amount of information on the 48 or 96-
hour TLm values of these compounds Unfortu-
nately, information is still lacking on their long-
term effects at sublethal levels on the productivity
of economically important marine species.
The extreme sensitivity of marine crustaceans,
such as crabs, lobsters, and shrimp, to the array of
insecticides is to be expected because of their phy-
logenetic relationship with terrestrial arthropods
In general, shrimp are also much more sensitive
than fish or oysters to the other pesticides This fact
and their economic importance make shrimp a
valuable yardstick for establishing safe levels of
pesticides that might be expected as toxicants in
the marine environment
A much broader spectrum of pesticide pollutants
can be anticipated in the fresh water (salinity
<0 5 %0) zones of tidal estuaries. Fresh water
criteria listed in another section will apply under
these circumstances.
Recommendation: The pesticides are grouped accord-
ing to their relative toxicity to shrimp, one of the most
sensitive groups of marine organisms Criteria are based
on the best estimates in the light of present knowledge
and it is expected that acceptable levels of toxic ma-
terials may be changed as the result of future research.
Pesticide group A.—The following chemicals are
acutely toxic at concentrations of 5 /ig/1 and less. On
the assumption that Koo of this level represents a rea-
sonable application factor, it is recommended that en-
vironmental levels of these substances not be permitted
to rise above 50 nanograms/1. This criterion is so low
that these, pesticides could not be applied directly in
or near the marine habitat without danger of causing
damage The 48-hour TLm is listed for each chemical
in parts per billion (/ig/l)
Organochloride pesticides
Aldrin 0 04
BHC 2 0
Chlordane 2 0
Endrin 0 2
Heptachlor 0 2
Lindane 0 2
DDT 0.6
Dieldrin 0.3
Endosulfan 0.2'
Melhoxychlor 4 0
Perthane 3 0
TDE 3.0
Toxaphene 3.0
Organophosphorous pesticides
Coumaphos 2 0 Naled 3 0
Dursban 3.0 Parathion I 0
Fenthion 0 03 Ronnel 5.0
Pesticide group B.—The following types of pesticide
compounds generally arc not acutely toxic at concen-
trations of 1 mg/1 or less. It is recommended that an
application factor of 4oo be used and, in the absence of
acute toxicity data that environmental levels of not
more than 10 Mg/1 be permitted
Arsenicals
Botanicals
Carbamates
2,4-D compounds
2,4,5-T compounds
Phthalic acid compounds.
Triazine compounds
Substituted urea compounds
Other pesticides.—Acute toxicity data are available
for approximately one hundred technical grade pesti-
cides in general use not listed in the above groups These
chemicals either are not likely to reach the marine en-
vironment, or, if used as directed by the registered
label, probably would not occur at levels toxic to marine
biota It is presumed that criteria established for these
chemicals in fresh water will protect adequately the
marine habitat It should be emphasized that no un-
listed chemical should be discharged into the estuary or
coastal water without preliminary biossay tests and the
establishment of an adequate application factor
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Heavy Metals: Heavy metal salts in solution
may constitute a very serious form of pollution
because they are stable compounds, not readily
removed by oxidation, precipitation, or any other
natural process. A characteristic feature of heavy
metal pollution is its persistence in time as well as
in space for years after the pollutional operations
have ceased.
The number of substances that may be described
as "poisonous" is very large and they vary enor-
mously in the degree of their effect. For man and
other air-breathing animals, the threshold dose of
a toxic material generally means the maximum
quantity that can be taken without causing death
For aquatic animals living in a water environment
containing a toxic substance, the situation is some-
what different. Instead of receiving an absolute
quantity at one time, they are being continually
exposed to a given concentration of the toxtc mate-
rial This is similar to a man regularly drinking
water containing lead or breathing air containing a
noxious gas or vapor It is not surprising, therefore,
that the student of pollution problems turns his at-
tention toward the concentration of the poison he
is investigating and the manner in which the effect
is related to this, rather than to the absolute
amount required to harm or kill Animals have the
ability to eliminate poisons at least to some degree
or even to destroy them Their ability to do this at
a rate permitting survival depends on the concen-
tration of the toxic material to which they are
exposed
One of the characteristics of living cells is their
ability to take up elements from a solution against
a concentration gradient This is perhaps most ob-
vious for marine organisms, especially for auto-
trophic algae which obtain all their nutrients di-
rectly from seawater The ability of marine
organisms to concentrate elements above that level
found in their environment has been recognized
for some time. The following points should be
noted in relation to their concentrating ability
(1) All elements are concentrated to a degree
with the exception of chlorine, which is rejected,
and sodium, which is weakly rejected. The concen-
tration factors are of the order of one for bromine,
fluorine, magnesium, sodium, and sulfur, and
higher for all other elements
(2) Among cations (including metallic ele-
ments such as iron, which may exist as colloids in
the sea), the order of affinity for living matter is,
generally tetravalent and trivalent elements>di-
valent transition elements> divalent group II-A
metals>univalent group I metals. The tetravalent
and trivalent subgroup have rather different affini-
ties for plankton and brown algae.
plankton: Fe>Al>Tl>Cr, Si>Ga
brown algae: Fe>La>Cr>Ga>Li>Al>Si
Similar differences are found between these orga-
nisms in their affinities for the di/alent transition
metals
plankton: Zn>Pb>Cu>Mn>Co>Ni>Cd
brown algae. Pb>Mn>Zn>Cu, Cd>Co>Ni
Of interest is the affinity of both organisms for lead,
which has no known biological function.
It is clear that the heavier elements in these
groups tend to be more readily taken up than the
lighter ones, which may be connected with their
greater, ease of polarization.
(3) The order of affinity of living matter for
anions is
nitrate > trivalent anions > divalent anions >
univalent anions
It is probable that most polyvalent metallic ele-
ments are more or less chelated by organic matter
The main features of the uptake of ions by cells
can be accounted for by assuming that another
process operates apart from simple diffusion This
process is called active uptake and is closely linked
with metabolic activities within the cell The meta-
bolic processes provide the energy necessary for
the uptake against a concentration gradient Active
uptake has a larger temperature coefficient than
does uptake by diffusion In long-term experi-
ments, the effect of temperature is probably com-
plicated by increased rates of growth, cell division,
and so on Active uptake requires oxygen and oc-
curs only in cells which are respiring freely Sub-
stances which inhibit respiration also inhibit up-
take of ions The rate of uptake of ions may be
limited either by the rate of exchange at the cell
membrane or by bulk phase diffusion inside the
cell The former is usually limiting for ions present
at low external concentration and the latter for
ions at high external concentrations. It has been
suggested that bulk phase diffusion limits the rates
of uptake of most cations There appears to be at
least two active transport systems in addition to
the diffusion processes A large number of theories
have been advanced to explain active transport
One of the most popular is the carrier hypothesis.
Accordingly, the ions are transported across mem-
branes as chelates with metabolically produced or-
ganic molecules.
Uptake by invertebrate animals.—The most
primitive animals, the unicellular protozoa, take
up ions from solution by diffusion in the same
ways as do algae Many marine species have
vacuoles and these are able to open at intervals
and extrude fluid from the cell. The vacuole regu-
-------
lates the osmotic pressure of the cell and thus con-
trols its volume.
Multicellular invertebrate animals can be di-
vided into two groups as far as uptake is con-
cerned: those with permeable integuments and
those without. The majority of marine inverte-
brates (colenterates, annelids, mollusks, and
echinoderms) have soft bodies with permeable
integuments through which ions can diffuse freely
In this situation, the body fluid or blood is quite
similar to sea water in composition The gills of
mollusks are coated with a layer of complex carbo-
hydrate sulfates which may function as ion ex-
changers The gills of marine Crustacea, which
have hard impermeable carapaces, are fully per-
meable to water and salts.
Mode of toxic action.—An element is satd to be
toxic if it injures the growth or metabolism of an
organism when supplied above a certain concen-
tration. All elements are toxic at high concentra-
tions and some are notorious poisons even at low
concentrations For example, the essential micro-
nutrient, copper, which is a necessary constituent
of all organisms, is highly toxic at quite small con-
centrations The other essential micronutrients are
also toxic when supplied in excess, though not all
in such striking fashion There is an optimum range
of concentration, which is sometimes quite narrow,
for the supply of each element to each organism
When excessive amounts of an element are fed
to an organism, they frequently cause death The
usual measure of the amount required to cause
death is called the LD50 This is the amount which,
when fed to each individual in a population, kills
half of the population. The LD5() is an imprecise
measure unless it is qualified by specifying
(1) The chemical state of the element.
(2) The means of feeding
(3) The age or developmental stage of the
organism
(4) The time elapsed between feeding and
death
The most important mechanism of toxic action
is thought to be the poisoning of enzyme systems.
The more electronegative metals, notably copper,
mercury, and silver, have a great affinity for amino,
immo, and sulfhydryl groups which are doubtless
reactive sites on many enzymes These metals are
readily chelated by organic molecules We thus
have discovered attempts to correlate metal toxi-
cities with such factors as their electronegativities,
the insolubility of their sulfides, or the order of
stability of their chelated derivatives.
(1) Order of electronegativities of some diva-
lent metals Hg>Cu>Sn>Pb>Ni>Co>
Cd>Fe>Zn>Mn>Mg>Ca>Sr>Ba
(2) Order of stability products of the sulfides:
Hg>Cu>Pb>Cd>Co>Ni>Zn>
Fe>Mn>Sn>Mg>Ca
(3) Order of stability of chelates: Hg>Cu>
Ni >Pb>Co>Zn>Cd>Fe>Mn>Mg>
Ca
It appears likely that all the divalent transition
metals, as well as the other electronegative metals,
that form insoluble sulfides, such as Ag, Mo, Sb,
Tl, and W, are poisons by virtue of their reactivity
with proteins and especially with enzymes. In view
of the large number of enzymes in living cells, the
variations in toxicity indicated above are hardly
surprising Studies have shown that metals giving
rise to similar toxic effects may be acting on quite
unrelated enzymes and also many more atoms of
metal are absorbed by an inactivated enzyme than
are required to block the reactive sites Other
modes of toxic action are.
(1) Substances behaving as antimetabolites.
This might be arsenate and chlorate occu-
pying sites for phosphates and nitrates,
respectively (Fluoride, borate, bromate,
permanganate, antimonate, selenate, tellu-
rate, tungstate, and beryllium.)
(2) Substances forming stable precipitates or
chelates with essential metabolites. (Al,
Be, Sc, Ti, Y, Zr, reacting with phosphate,
Ba with sulfate, or Fe with ATP.)
(3) Substances catalyzing the decomposition of
essential metabolites. (La and other lan-
thamde cations decompose ATP.)
(4) Substances combining with the cell mem-
brane and affecting its permeability. (Au,
Cd, Cu, Hg, Pb, U ) These elements may
affect transport of sodium, potassium,
chlorine, or organic molecules across mem-
branes or even rupture them.
(5) Substances replacing structurally or elec-
trochemically important elements in the
cell and then failing to function (Li replac-
ing Na, Cs replacing K, or Br replacing
CI )
Metal-organic compounds may be either more
toxic than the metal ion (ethyl mercuric chloride)
or much less so (cupnc ion and copper salicyl-
aldoxime)
Silver.—Silver is present in seawater in a con-
centration of about 0.0003 mg/1 It is found in
marine algae at concentrations up to 0 25 mg/1
and in marine mammals in the range of 1 to 3 mg/1
(Vinogradov, 1953). It is highly toxic to plants
and mammals.
Arsenic.—Arsenic is found to a small extent in
nature in the elemental form. It occurs mostly in
the form of arsemtes of true metals or as pyrites
-------
Its major commercial use is for pesticides (insects,
weeds, fungi). Arsenic is cumulative in the tissues
of many organisms and, therefore, it eventually
exerts its effects even though the environmental
level is low. It has been demonstrated to be a pos-
sible carcinogen in water.
Arsenic is found in seawatcr at a concentration
of about 0 003 mg/1. It has been found in marine
plants at concentrations up to 30 mg/1 and is high-
est in the brown algae It is found in marine ani-
mals in a range of 0.005 to 0 3 mg/1. It is accumu-
lated by coelenterates, some mollusks, and
crustaceans (Vinogradov, 1953) It is moderately
toxic to plants and highly toxic to animals especi-
ally as AsH3.
Arsenic trioxide, which also is exceedingly toxic,
was studied in concentrations of 1.96 to 40 mg/1
and found to be harmful to fish or other aquatic
life Work by the Washington Department of Fish-
eries (1944) on pink salmon has shown that at a
level of 5 3 mg/1 of As^Oj for 8 days was extremely
harmful to this species Ellis (1937), using the
same compound on mussels at a level of 16 mg/1,
found it to be quite lethal in 3 to 16 days. Surber
and Meehan (1931) carried out an extensive study
on the toxicity of As2Oj to many different fish food
organisms Their results indicated that important
fish food organisms can tolerate an application rate
of 2 mg/1 of Asj03 The amount actually in the
water is considerably less
Cadmium.—The elemental form of cadmium
is insoluble in water. It occurs largely as the sulfide
which is often an impurity in zinc ores
Cadmium is found in seawater at a level of less
than 0 08 mg/1 Its level in marine plants is ap-
proximately 0 4 mg/1, while in marine animals a
range of 0 15 to 3 mg/1 has been found. It is low-
est in the calcareous tissues and is accumulated
within the viscera of the mollusk, Pecten novazet-
landicae (Brooks and Rumsby, 1965) Cadmium
is moderately toxic to all organisms and it is a
cumulative poison in mammals
Cadmium is used widely industrially to alloy
with copper, lead, silver, aluminum, and nickel It
is also used in electroplating, ceramics, pigmenta-
tion, photography, and nuclear reactors. Cadmium
salts sometimes are used as insecticides and anti-
helminthics The chloride, nitrate, and sulfate of
cadmium are highly soluble in water. The carbo-
nate and hydroxide are insoluble, thus cadmium
will be precipitated at high pH values.
Most quantitative data on the toxicity of cad-
mium are based on specific salts of the metal Ex-
pressed as cadmium, these data indicate that the
acute lethal level for fish varies from about 0 01 to
about 10 mg/1 depending on the test animal, the
type of water, temperature, and time of exposure.
Cadmium acts synergistically with other substances
to increase toxicity. Concentrations of 0.03 mg/1
in combination with 0.15 mg/1 zinc causes mor-
tality of salmon fry (Hublou, et al., 1954).
Pringle (in press), in a study of adult American
Eastern oysters, Crassostrea vtrgimca, found an 8-
week TLm value of 0 2 mg/1 of Cd'+[Cd(N03)2]
and a 15-week TLm value of 0 I mg/1.
The most obvious effect, in addition to lethality,
was lack of shell growth. A similar study on the
clam, Mercenaria mercenaria, indicated that a
much longer period of exposure at the same con-
centration was required to kill half of the test
organisms.
Chromium.—Chromium is found in seawater at
a concentration of 0.00005 mg/l. Marine plants
contain approximately 1 0 mg/l while marine ani-
mals contain chromium within a range of 0.2 to
1 0 mg/l Chromium compounds may be present
in wastes from many industrial processes or they
may be discharged in chromium-treated cooling
waters The toxicity of chromium varies with the
species, temperature, PH, its valence, and synergis-
tic or antagonistic effects (especially with hard-
ness). Most evidence points to the fact that under
long-term exposure the hexavalent form is no more
toxic towards fish than the tnvalent form. Doudor-
off and Katz (1953), studied the effect of K2Cra07
on mummichaugs and found that they tolerated a
200 mg/l level in sea water for over a week
The effects of hexavalent chromium on photo-
synthesis by the giant kelp, Macrocystis pyrijera,
were as follows at 1 mg/l chromium, photosyn-
thesis was not diminished by 2 days contact. It
was reduced 10 to 20 percent by 5 days contact
and 20 to 30 percent after 7 to 9 days The con-
centration of chromium required to cause a 50-per-
cent mactivation of photosynthesis in 4 days was
estimated at 5 mg/l (Clendenning and North,
1958. 1960, North and Clendenning, 1958, 1959).
Haydu (unpublished data) studied oyster mor-
talities and his results point out the long-term ef-
fects of low concentrations of chromium, molybde-
num, and nickel. The levels of all three metals were
in the range of 10 to 12 /*g/l over a 2-year period.
In addition, his data indicated that there were sea-
sonal variations The mortalities at these levels in«
creased with an increase in temperature. Approxi-
mately 63 to 73 percent of the mortalities occurred
in the period of May through July, perhaps due to
increased physiological activity (increased feeding
and higher pumping rates)
This study substantiates the available evidence
indicating that as the environmental level of these
metals increases, the mgestion-elimination balance
-------
is upset, causing accumulation to take place
Raymont and Shields (1964), in studies with
the small prawn, Leander squiUa, found a thresh-
old level of a little less than 5 mg/1 Cr Thus, at
chromium concentrations ranging from 10 to 80
mg/1 Cr, 100-percent mortality occurred m 1
week, at 5 mg/1 Cr no deaths occurred in 1 week
although a few animals died over the subsequent
21 days. Larger prawns of the same species ap-
peared to be considerably more resistant to chro-
mium poisoning. The threshold was about 10 mg/1
Cr Raymont and Shields in additional experiments
on the toxicity of chromium to crustaceans (the
shore crab, Carcinus maenas), indicated that chro-
mium concentrations above 50 mg/1 (Na2CrOH)
were definitely toxic for a period of exposure of
12 days At 60 mg/1 Cr, 50-percent mortality oc-
curred after 12 days. At 40 mg/1 Cr, 9 percent
died within 12 days, while at 20 mg/J, an 8-percent
mortality was observed. In studies on the marine
polychaete worm, Nereis virens, these same inves-
tigators working in the range of 2 to 10 mg/1 Cr
found that there was heavy mortality with all solu-
tions in 2 to 3 weeks. The threshold of toxicity ap-
pears to be at about 1 0 mg/1 Cr level
Pringle (in press), in experiments using a well-
controlled, flow-through system and chromium
concentrations of 0 1 and 0 2 mg/1 (NazCr207),
showed the average weekly mortality to be ap-
proximately 1 percent over a 20-week period. This
was about the same as that for the sea water
controls.
Copper.—Copper is found in seawater at a level
of 0 003 mg/1 It is found in marine plants at
about 11 mg/1, while marine animals are found to
contain 4 to 50 mg/1 It is accumulated by some
sponges and is essential for the respiratory pig-
ment in the blood of certain annelids, Crustacea,
and mollusks In excess, it is highly toxic to algae,
seed plants, and to invertebrates and moderately
toxic to mammals Copper is not considered to be
a cumulative systemic poison like lead or mercury
The toxicity of copper to aquatic organisms
varies significantly not only with the species but
also with the physical and chemical characteristics
of the water Copper acts synergistically with zinc,
cadmium, and mercury, yet there is a sparing
action with calcium.
Barnacles and related marine fouling organisms
were killed in 2 hours by 10 to 30 mg/1 copper
Clarke (1947) showed that the mussel, Mytilus
edulis, was killed in 12 hours by 0.55 mg/1. Lob-
sters transferred to tanks lined with copper after
living in aluminum, stainless steel, and iron tanks
for 2 months, died within 1 day Copper is concen-
trated by plankton from surrounding water in
ratios of 1,000 to 5,000 or more (Krumholz and
Foster, 1957)
Concentrations of copper above 0.1 to 0.5
mg/1 were found to be toxic to oysters by Galtsoff
(1932) The 96-hour TLm for oysters was esti-
mated at 1.9 mg/1 (Fujiya, 1960). Oysters cul-
tured in waters containing 0.13 to 0.5 mg/1 ac-
cumulated copper in their tissues and became unfit
as a food substance. Pringle (in press) found the
soft clam, Mya arenaria, extremely sensitive to
copper. At a concentration of 0 5 mg/1, 100-per-
cent mortality took place in 3 days. Using a 0.2
mg/1 concentration at 10 and 20 C, all clams died
within 23 days at the lower temperature, while at
the higher temperature all succumbed in 6 to 8
days When 0 1 mg/1 Cu at 20 C was used, all
animals died in 10 to 12 days. Raymont and
Shields (1964) in studies with the marine poly-
chaete worm Nereis, showed that a concentration
of 1 5 mg/1 Cu was lethal in 2 to 3 days, and con-
centrations exceeding 0.05 mg/1 Cu were lethal in
approximately 4 days.
Clendenning and North (1958, 1960) and
North and Clendenning (1958, 1959) evaluated
the effect of copper (from the chloride and sulfate
salts) on the rate of photosynthesis of the giant
kelp, Macrocystis pyrifera With 0.1 mg/1 of cop-
per, net photosynthesis was inhibited by 50 percent
in 2 to 5 days and 70 percent in 7 to 9 days. Visi-
ble injury appeared in 10 days. Copper was
slightly less toxic than mercury but more so than
nickel, chromium, lead, or zinc Marvin, Lansford,
and Wheeler (1961) found 0 05 mg/1 Cu toxic to
Gymnodmium breve (red tide organism).
Mercury.—Mercury is found in seawater at a
level of 0 00003 mg/1 It is found in marine plants
at approximately 0.03 mg/1.
Irukayama (1966) reported on a mercurial pol-
lution incident in Japan, which was first recognized
in 1953 A severe neurological disorder resulted in
the area of Minamata Bay as a result of eating fish
and shellfish from these waters. Many species of
animals including waterfowl were succumbing to
the "disease" called Minamata disease. Clinical
features were cerebellar ataxia, constriction of vis-
ual fields, and dysanthia Pathological findings
were regressive changes in the cerebellum and
cerebral cortices. Investigation through 1965 sug-
gested that the main cause was the spent factory
waste of the Kanose Factory upstream from the
Minamata Bay area. Methyl mercury compounds,
waste byproducts from the acetaldehyde synthesis
process, were being discharged and concentrated
especially in shellfish
Ukeles (1962) made a study of pure cultures of
-------
marine phytoplankton in the presence of toxicants.
One of the toxic materials used was lignasan
(ethyl mercury phosphate) a bactericide-fungi-
cide. She found lignasan to be lethal to all species
at 0 06 mg/l and 0.0006 was the highest level used
not causing drastic inhibition of growth.
Clendenning and North (1960) and North and
Clendenning (1958) found that 0.5 mg/l of mer-
cury added as mercuric chloride caused a 50-per-
cent inactivation of photosynthesis of the giant
kelp, Macrocystis pyrijera, during a 4-day expo-
sure A concentration of 0.1 mg/l caused a 15-
percent decrease tn photosynthesis in 1 day and
complete inactivation in 4 days. Mercury was more
toxic than copper, hexavalent chromium, zinc,
nickel, or lead. For phytoplankton, the minimum
lethal concentration of mercury salts has been re-
ported to range from 0.9 to 60 mg/l of mercury
(Hueper, 1960). The toxic effects of mercury salts
are accentuated by the presence of trace amounts
of copper (Corner and Sparrow, 1956).
Lead.—Lead is found as a local pollutant of
rivers near mines and from the combustion of
leaded gasolines. The lead concentration in sea-
water is in the order of 0 00003 mg/l. It is found
in marine plants at a level of approximately 8 4
mg/l. Residues in marine animals reach a concen-
tration in the range of 0.5 mg/l. It is highest in
calcareous tissue.
Wilder (1952) found that lobsters died within
20 days when kept in lead-lined tanks, while in
steel-lined and other types of tanks, they survived
for 60 days or longer.
North and Clendenning (1958) found that lead
was less toxic to the giant kelp, Macrocystis pyri-
feia, than mercury, copper, hexavalent chromium,
zinc, or nickel
Pnngle (unpublished data), in studies on the
effects of lead on the Eastern oyster, Crassostrea
virginica, found a 12-week TLm value of 0.5 mg/l
and an 18-week TLm value of 0.3 mg/l. Concen-
trations of 0 1 to 0 2 mg/l induced noticeable
changes in mantle and gonadal tissue under 12
weeks of exposure.
Nickel.—Nickel is found in sea water in a con-
centration of about 0 0054 mg/l. Marine plants
contain up to 3 mg/l and this may be higher in
plankton. Marine animals contain levels in the
range of 0.4025 mg/l. Nickel pollution is caused
by industrial smoke and other wastes. It is very
toxic to most plants but less so to animals Haydu
(unpublished data), in long-term studies with oys-
ters, found that a level of 0.121 mg/l nickel
caused considerable mortality.
Zinc.—Zinc is found in sea water in a concen-
tration of 0.01 mg/l. Marine plants may contain
up to 150 mg/l of zinc. Marine animals contain
zinc in the range of 6 to 1500 mg/l. It is accumu-
lated by some species of coelenterates and mol-
lusks Speer (1928) reports that very small
amounts of zinc are dangerous to oysters.
Clendenning and North (1960) and North and
Clendenning (1958) tested the effect of zinc sul-
fate on the giant kelp, Macrocystis pyrifera. Four-
day exposure to 1.31 mg/I of zinc showed no ap-
preciable effect on the rate of photosynthesis, but
10 mg/l caused a 50-percent inactivation of kelp.
Other Toxicants
Ammonia-ammonium compounds.—Ammonia
is found m the discharge of many industrial wastes.
It has been shown that at a level of 1.0 mg/l NH,,
the ability of hemoglobin to combine with oxygen
is impaired and fish may suffocate. Evidence indi-
cates that ammonia exerts a considerable toxic ef-
fect on all aquatic life within a range of less than
1.0 mg/l to 25 mg/l, depending on the pH and
dissolved oxygen level present.
Cyanides.—Hydrocyanic acid or hydrogen cya-
nide and its salts, the cyanides, are important in-
dustrial chemicals The acid and its salts are
extremely poisonous.
Hydrogen cyanide is largely dissociated at pH
levels above 8 2 and its toxicity increases with a
decrease in pH. The toxic action of cyanides in-
creases rapidly with a rise in temperature.
Fish can recover from short exposure to con-
centrations of less than 1 0 mg/l (which seems to
act as an anaesthetic) when removed to water free
of cyanide. They appear to be able to convert cya-
nide to thiocyanate, an ion that is not inhibitory
on the respiratory enzymes. Complex cyanides
formed by the reaction of CN with zinc or cad-
mium are much more toxic. However, the reaction
between CN and nickel produces a cyanide com-
plex less toxic than the CN itself at high pH levels.
Sulfides.—Sulfides in water are a result of the
natural processes of decomposition, sewage, and
industrial wastes such as those from oil refineries,
tanneries, pulp and paper mills, textile mills,
chemical plants, and gas manufacturing facilities.
Most toxicity data available are based on fresh
water fish. Concentrations in the range of less than
1 0 mg/l to 25 0 mg/l are lethal in 1 to 3 days.
Fluorides.—Fluorides are present in varying
amounts in the earth's crust They are used as in-
secticides as well as in water treatment and many
other uses While normally not present in industrial
wastes, they may be present in trace or higher con-
centrations due to spillage. Data in fresh water in-
dicate that they are toxic to fish at concentrations
higher than 1.5 mg/l.
Detergents and surfactants.—During the past
-------
twenty years, synthetic detergents have replaced
a majority of the soap products. Concern about
their importance in pollution was heightened by
the visible evidence of their foaming in the Na-
tion's waterways. Their toxicity to the aquatic
fauna has been very extensively studied, but for
the most part it is difficult to establish safe criteria
because of the varying conditions of the tests. Rela-
tively little bioassay work on their effects on marine
biota has been published, but it is indicated that,
unlike soap, detergents are more toxic m highly
saline water than they would be in the fresh water
areas of tidal estuaries (Eisler, 1965, Eisler and
Derrel, 1966).
The 96-hour TL,,, values of an ABS detergent
to five species of marine fish ranged from 7 to
22 mg/1 (Eisler, 1965). Marine kelp were more
sensitive and photosynthesis was inhibited 50 per-
cent after 96-hour exposures to about 1.0 mg/1
Pathogenic organisms.—Oysters, clams, and
mussels have a demonstrated ability to accumulate
microorganisms, including bacteria and viruses,
from their aquatic environments and to serve as
a vehicle for the transmission of these micro-
organisms to their consumers (U.S. DHEW, 1956,
1958, 1962, 1965a; Liu, et al, 1967).
Controls to prevent the transmission of disease
through this route have been provided in, the
United States through the National Shellfish Sani-
tation Program (NSSP) administered by the Pub-
lic Health Service, Department of Health, Educa-
tion, and Welfare on the behalf of the interested
State and Federal agencies and the shellfish in-
dustry (1965b). This program has established
bacteriological quality standards for those waters
from which shellfish are to be harvested for direct
marketing These standards, as described in the
NSSP Manual of Operation, should be observed
for those estuarine areas used for commercial pro-
duction of shellfish for direct marketing (U.S.
DHEW, 1965) The standards that are applied to
shellfish harvesting areas have been revised peri-
odically through the mechanism of a shellfish sani-
tation workshop held at 2 or 3-year intervals. As
these standards are revised so should the water
quality criteria be modified
Tar, gas, and coke wastes.—The distillation of
coal for the production of gas, coke, and tarry ma-
terials used in the manufacture of dyes and vari-
ous organic chemicals results in a watery waste
known as ammoniacal gas liquor, the disposal of
which can cause detrimental effects. Ammoniacal
gas liquor contains free ammonia, ammonium
salts, cyanide, sulfide, thiocyanate, and a variety
of aromatic compounds including pyridine,
phenols, cresols, xylenols, and aromatic acids
After treatment to remove ammonia, the waste is
called "spent gas liquor." Phenol or carboxylic
acid is the most abundant of its many phenolic
substances, probably the most dangerous to fish.
Phenolic substances are also present in materials
used in road surfacing, sheep dips, and many in-
dustrial wastes such as those associated with the
manufacture of plastics, dyes, and disinfectants.
Gas liquor, discharged untreated to a stream, has
an extremely high oxygen demand, many times
greater than that of sewage. These various groups
of organic substances produce a variety of effects
on fish varying from intoxication and anaesthesia
to paralysis and death.
Pure compounds representative of these groups
found in such coal tar wastes have been shown to
be toxic in ranges of 2 to 75 mg/1 for cresols and
0 1 to 50 mg/1 for phenols, for fresh water fish
and lower aquatic life.
Petroleum refining and petrochemical wastes.—
The volatile components of petroleum consist
mainly of aliphatic hydrocarbons. In addition to
paraffins and olefins, some petroleums contain
relatively high percentages of naphthenes and
aromatic hydrocarbons. The less volatile fractions
of petroleum are used as fuels, lubricants, and
construction materials (asphalt). These substances
are somewhat more irritating to the skin and some
are carcinogenic, but less so than coal tar products
Pulp and paper manufacturing wastes.—The
types of pulp produced and pulping technology
have undergone considerable change in the last
20 years and the trend continues. Modern pulp-
mills are geared to produce a variety of pulp
grades due to the increasing demands for specialty
products The characteristics of the waste waters
from these specialty pulp grades can vary con-
siderably. An example of this can be seen in the
BOD loadings of the following sulfite grade pulps
produced in a west coast mill:
Paper making—130 lb BOD/ADT (air dry
ton).
Alpha hardwood—300 lb/ADT.
F AC-SAC—450 lb/ADT.
The major pulping processes include kraft, sul-
fite, semichemical, and nonchemical such as
groundwood. The kraft process accounts for ap-
proximately 75 percent of the total pulp produc-
tion in the United States. The number of mills
using the sulfite process are declining, some are
being converted to the kraft process.
From the standpoint of water pollution, kraft
and sulfite mills are of great significance. The prin-
cipal problems associated with pulpmil! wastes are
toxicity, depressed DO's, and slime growths. Clear-
cut cases of acute toxicity attributable to pulpmill
wastes in modern times seldom exist except when
-------
spills or other accidents occur. It is much more
common to encounter problems related to slime
growths, depressed DO's, and to long-term or
chronic effects on the biota.
A substantial portion of pulpmill wastes includ-
ing the toxic components are very amenable to
microbial degradation. In one study, kraft mill
wastes were found to be nontoxic to oysters at a
dilution of 1:20 when the BOD of the waste was
reduced by 80 percent employing biological treat-
ment In a similar study, the toxipity of kraft
wastes to silver salmon was found to diminish pro-
portionally to the degree of BOD reduction above
50 percent, again using biological treatment The
results of a recent study by scientists of the Inter-
national Pacific Salmon Commission indicate a
fairly close relationship between BOD reduction
and decrease in the toxicity of kraft wastes They
found no apparent toxicity to salmon when the
BOD was reduced by 65 percent While similar
studies have not been made with sulfite liquors,
there is some evidence that the toxic components
of this waste are also degradable It is important to
recognize that the biological mechanism or degra-
dation involved in secondary treatment is essen-
tially similar to that in receiving waters Given
sufficient time, the process of degradation of the
toxic components of pulp wastes also take place in
receiving waters.
Because of the great complexity and variability
of pulpmill wastes, it is difficult to find a satisfac-
tory expression for concentration. Attempts have
been made to relate toxicity to BOD, COD, total
solids, PBI (Pearl Benson Index—a measure of
the lignin content of pulp wastes), and various
reference animals There is a general relationship
with all of these criteria, i e., the higher the values,
the greater the toxicity. Pulpmill dosages or dilu-
tions have been used in bioassays on the basis of
applied initial BOD. The response of the test ani-
mals has been found to vary considerably to given
concentrations of applied BOD even from the
wastes of the same mill. This would indicate that
the concentration of toxicants in the total biolog-
ically amenable fraction is subject to considerable
variation. This would not only explain the lack of
a good relationship between the toxicity and initial
BOD, but it would also explain why, on the other
hand, there can coexist a good relationship be-
tween BOD reduction and reduction in toxicity.
The latter is subject to degradation regardless of
the proportions of toxicants and the other to bio-
degradable substances
The shortcomings of BOD as an expression of
the concentration of toxicity would seem to be
equally applicable to the PBI test. This test has
been recommended as a measure of SWL (sulfite
waste liquor) concentration. It measures the
lignins m SWL which constitute an appropriate
substance for tracing in receiving waters and for
analysis due to their stability and high concentra-
tions As indicated earlier, critical tests to deter-
mine the relationship between BOD reduction and
reduction in toxicity have not been conducted with
SWL. Nevertheless, there is sufficient evidence to
indicate that the toxic components of SWL also
reside in the biodegradable fraction and are also
degradable The composition of SWL in receiving
waters at different distances from the point of dis-
charge would therefore differ even though similar
PBI values may occur. The toxicity of fresh SWL
at a PBI concentration of 50 mg/1 would be much
greater than of biologically stabilized SWL at the
same PBI concentration. There is clear indication
that further study of SWL toxicity and biodegrada-
tion is necessary
The toxicity of kraft and sulfite wastes to
aquatic life is amply reported in the literature
Deleterious effects produced by SWL (generally
considered less toxic than kraft wastes) are re-
ported from PBI values as low as 2.0 mg/1 for
oyster larvae to concentrations greater than 1,000
mg/1 for the adult clams Mya and Macoma. Long-
term bioassays with Pacific and Kumamoto
oysters, carried out at Oregon State University
using calcium-base SWL (10 percent solids),
showed no adverse effects at 50 mg/1 after 266
days of exposure Slightly deleterious effects were
noted at 100 mg/1, indicating maximum safe
limits lie between 50 to 100 mg/1 Continuing field
studies at Grays Harbor, Wash , support these
findings. In bioassays conducted in salt water by
the Washington State Department of Fisheries, sal-
mon exposed for 30 days to concentrations of ap-
proximately 500 mg/1 of 10-percent SWL showed
no apparent ill effects Herring eggs, on the other
hand, were adversely affected at concentrations
greater than 96 mg/1
The apparent tolerance level for salmon in salt
water using kraft wastes was found by the above
investigation to range from dilutions of 1:16 to
1 90 after 14 to 30 days of exposure Growth
studies conducted at Oregon State University by
the National Council for Stream Improvement
using raw kraft wastes in fresh water showed no
adverse effects to salmonid fishes after 3 to 5
weeks exposure in dilutions of 1:100. English (in
press), in his field studies of the English sole in
Puget Sound, reports a sustained and thriving
fishery in an area affected by SWL. Recent work
by the Federal Water Pollution Control Adminis-
tration (USDI 1967a) in Puget Sound showed
-------
damage to oyster larvae and developing English
sole eggs at concentrations greater than 10 mg/1
of 10-percent SWL. According to this report,
oyster growth and market condition is adversely
affected and phytoplankton productivity is in-
hibited at SWL concentrations over 50 mp,/l.
Determining the toxicity of complex wastes like
oil, refinery petrochemicals, and pulpmill wastes
presents a number of problems. For one thing,
they contain many known and, perhaps, equally
as many unknown toxic substances in small quan-
tities. The toxicological and other physical and
chemical characteristics can vary considerably dur-
ing any given day, in any given plant, due to
changes in processes, sources of supplies, and the
end product being produced. Considerable varia-
tion in effluent characteristics can occur even in a
1-day period. The resulting wastes from these in-
dustries contain upwards of several hundreds of
compounds representing a number of homologous
series of compounds from different organic groups.
This complexity is augmented by the treatment of
the wastes, as well as by the spectrum of products
manufactured from the complex starting material
used The relative ability to react biochemically
and to exert an oxygen demand is characteristic of
organic materials of such primary significance.
Many groups or series of compounds indicated
to be present in such wastes have been shown to
be toxic in varying degrees to aquatic life It is
extremely difficult at this time, however, to place a
concentration limit or set threshold criteria for
such complex systems and hence should be indi-
vidually bioassayed and their discharge managed
accordingly
Waterfront and boating activities.—Increasing
activities by commercial, military, and recreational
vessel operators raise the specter of introduction of
toxic materials in quantities sufficient to affect
marine organisms adversely. This is particularly
likely in the case of confined waters of small tidal
tributaries, lagoons, embayments, and other ma-
rine areas employed as harbors.
Toxic materials are used to prevent activities of
boring and fouling marine organisms Usually,
however, every effort is bent in the case of toxic
coatings to prevent rapid release of toxic materials
into the environment since rapid loss reduces ef-
fectiveness of such coatings and increases costs.
Some leaching is unavoidable—even necessary.
Thus, the presence in confined harbors of many
vessels whose bottoms are coated with toxic mate-
rials already presents hazards in some places This
would be especially true after spring "fitting out"
for small boats
Boatowners, boat and boatyard operators, fish-
ing and commercial pier and marina operators are
not especially noted for the care extended to
nearby waters Commonly, everything that can be
is flushed or jettisoned into the water. Purposeful
discharges are many—though perhaps decreasing
as emphasis on water pollution has increased.
Paint leaching, paint spillage, oil and gasoline
spillage, detergents, wood preservatives, ex-
hausted containers, metallic objects of all types
(zinc, copper, brass, iron, etc.), and other jetsam
contribute to contamination from these sources.
Except for confined areas where there are many
of these operations such as large shipyards, major
military and commercial anchorages, and large
and small boat anchorages, it is doubtful that tox-
icity from these operations is of serious proportions
in tidal waters at this point. As with other fouling
or contaminating activities of society, however,
efforts should be made to keep biological damage
from these sources to a minimum Some discharges
are controllable and should fall under the same
rules as industrial or community discharges. In the
case of large marinas, shipyards, or major anchor-
ages, requirements suggested elsewhere may have
to be applied Future research should include spe-
cific attention to this aspect.
Similar comments can be made about water-
front structures and port operations There is con-
siderable use of toxic materials in preservation of
wood, steel, and masonry structures used on ma-
rine waterfronts Discharge of toxic materials,
surfactants, petroleum products, other materials
and jetsam is common Similar recommendations
can be made for control and research as those for
boat, boatyard, and vessel operations.
Disposal of laboratory wastes.—The rapid
growth of marine sciences during the past decade
is reflected in an ever-increasing number of sta-
tions and laboratories engaged in the study of
various aspects of oceanography These institu-
tions are located along the entire coastline of the
United States: 28 on the Atlantic, 12 on the Gulf,
and 29 on the Pacific About 2,500 persons (in-
vestigators, students, technicians, and laboratory
assistants) are employed in these 66 establish-
ments (Hiatt, 1963)
The above number includes institutions operated
by Federal and State governments, by universities
or privately endowed concerns which receive their
main support from the government and national
foundations. Other laboratories, hospitals, and re-
search institutions operated by industrial concerns
for their specific needs are not included in this
total The laboratories range from small establish-
ments, with less than four investigators, to very
large institutions employing or providing research
space for 200 to 500 investigators.
-------
The types of research cover various fields of
biology, microbiology, experimental physiology,
biochemistry, chemistry, biophysics, molecular
biology, radiobiology, fishery biology, fishery man-
agement, and industrial research. Consequently,
the effluents discharged into estuarine and coastal
waters vary from ordinary household sewage to
mixtures containing an array of organic and inor-
ganic compounds, drugs, and radioactive isotopes.
The composition of these effluents cannot be pre-
dicted with certainty because the type of research
varies greatly from year to year. The laboratory
effluent is separated usually from the sea water
system, which as a rule has independent plumbing,
but is mixed with the domestic sewage and fre-
quently is discharged into natural waters. When
many scientific establishments are concentrated in
a relatively small area, the situation may become
serious. Chlorinated raw sewage entering the
harbor a short distance from shore may be caught
by a tidal eddy and for several hours circulate
close to the sea water intakes of several labora-
tories before it is carried out by tides
To maintain desired water quality requirements
for aquatic life, it is necessary to separate labora-
tory effluents from domestic sewage and provide
treatment that renders them harmless to aquatic
biota Under no conditions should highly toxic
chemical compounds or drugs be permitted to be
discharged into natural waters if toxic concentra-
tions of them can be detected by chemical and
physical methods.
Many marine laboratories are utilizing exotic
and endemic microorganisms, some pathogenic,
in research Extreme caution must be exercised to
prevent contamination of water by introduction
of biological materials which can harm marine
organisms
Laboratory administrators should be responsible
for the periodical examination of the toxicity of
the effluent discharged into natural waters by their
institutions
Recommendation: (1) Allowable concentrations of
metals, ammonia, cyanides, and sulfides should be
determined by the use of 96-hour TLn values and
appropriate application factors Preferably, the TLm
values should be determined by flow-through bioassays
in which environmental factors are maintained at levels
under which these materials are most toxic Tests
should utilize the most sensitive life stage of species M
ecological or economic importance in the area. Tenta-
tively, it is suggested that application factors should
be '/ioo for pesticides and metals, Via for ammonia,
Wo for cyanide, and for sulfides. ^
(2) There is evidence that fluorides are accumula-
tive in organisms It is tentatively suggested that
allowable levels should not exceed those for dnnking
water -
(3) The further dilution of wastes in marine waters
suggests that the adoption of criteria established for
detergents and surfactants in fresh water also will
protect adequately biota in the marine environment.
(4) Bacteriological criteria of estuarine waters
ultilized for shellfish cultivation and harvesting should
conform with the standards as described in the Na-
tional Shellfish Sanitation Program Manual of Opera-
tion These standards provide that'
(a) Examinations shall be conducted in accord-
ance with the American Public Health Association
recommended procedures for the examination of
sea water and shellfish
(b) There shall be no direct discharges of un-
treated sewage.
(c) Samples of water for bacteriological examina-
tion to be collected under those conditions of time
and tide which produce maximum concentration of
bacteria.
(d) The cohform median MPN of the water does
not exceed 70/100 ml, and not more than 10 percent
of the samples ordinarily exceed an MPN of 230/
100 ml for a 5-tube decimal dilution test (or 330/
100 ml where the 3-tube decimal dilution test is
used) in those portions of the area most probably
exposed to fecal contamination during the most un-
favorable hydrographic and pollution conditions
(e) The reliability of nearby waste treatment
plants shall be considered in the approval of areas
for direct harvesting
(5) It is also essential to monitor continuously waste
from tar, gas, and coke, petroleum refinery, petrochemi-
cal, and pulp and paper mill operations. They all
produce complex wastes of great variability, not only
from facility to facility, but also from day to day
This should be done on an individual basis with bio-
assays These tests should be made at frequent inter-
vals to determine TLm values as described for other
wastes. For the more persistant toxicants, an applica-
tion factor of Vioo should be used while for unstable or
biodegradable materials an application of Mm is tenta-
tively suggested
(6) Concentration of other materials with noncumu-
lative toxic effects should not exceed Ho of the 96-hour
TLo, value For toxicants with cumulative effects, the
concentrations should not exceed Mm and Vim for the
above respective values
When two or more toxic materials that have additive
effects are present at the same time in the receiving
water, some reduction in the permissible concentrations
as derived from bioassays on individual substances is
necessary The amount of reduction required is a func-
tion of both the number of toxic materials present and
their concentrations with- respect to the derived per-
missible concentration An appropriate means of assur-
ing that the combined amounts of the several substances
do not exceed a permissible combination for the mix-
ture is through the use of following relationship
(£+£¦ •¦+£*')
Where C., Ct, . C0 are the measured concentra-
tions of the several toxic materials in the water and
L., Lb L, are the respective permissible concen-
trations (limits) derived for the materials on an in-
dividual basis Should the sum of the several fractions
exceed one, then a local restriction on the concentra-
tion of one or more of the substances is necessary
-------
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FISH, OTHER AQUATIC LIFE, AND WILDLIFE
I INTRODUCTION
Research aimed at improving and main-
taining tue environment of aquatic
organisms is not limited in objective to
the benefit of these organisms. It is
meant to provide adequate supplies of
aquatic life and wildlife for man's use
Thus, in approaching this problem, the
question should be, "What is the quality
and quantity of aquatic resources that
man now has and should have in the future,
and how can these resources be restored,
protected, maintained, and enhanced"? "
To meet these problems, the determination
of water quality requirements for aquatic
life is of outstanding importance. If a
fresh water is suitable and safe for aquatic
organisms, it can be rendered safe for
human consumption by water treatment
methods now in existence. If fresh water
is satisfactory for aquatic plants and
aquatic organisms, it will m most cases
be satisfactory for livestock, wildlife, and
irrigating crops. If a water is satisfactory
for aquatic life, it will in general be
aesthetically pleasing and suitable for most
recreational uses. Water suitable for
aquatic life should be entirely suitable for
industry, navigation, and power. The
establishment of water quality require-
ments for the protection of aquatic life
resources is the key to the protection of
most uses made of water.
In the past, only a small percentage of the
available funds for controlling water
pollution was utilized for the determination
of water quality requirements for aquatic
life and wildlife. Biological investigations
were not carried out in proportion to their
importance. Very little attention was
given to determining environmental re-
quirements of aquatic organisms and the
concentrations of potential toxicants that
are not harmful in the aquatic environment.
Asa result, the volume of work was
entirely inadequate to meet the problem.
Because of deficiencies in equipment and
personnel, studies were short term and
_dealt largely with acute toxicity. Little
research was done on determining the
long-term effects of pollution, or levels
of potential toxicants that are not harmful
in the aquatic environment.
It was obvious to the Committee from the
onset of their task that present knowledge
is inadequate for establishing definitive
and conclusive water quality requirements
for aquatic life and wildlife. This
deficiency became more obvious as the
preparation of the Subcommittee Report on
Water Quality Criteria progressed and the
present consensus is that the elucidation of
water quality requirements has barely
begun. Only research that is broad in
scope, pertinently oriented, carefully
designed, and adequately financed will
supply the information essential for the
protection of fish, other aquatic life, and
wildlife.
II RESEARCH APPROACHES, METHODS,
AND PROCEDURES
Short and long-term bioassays are presently
the only tools available for determining the
toxicity of particular pollutants to aquatic
life, other approaches are needed to provide
an understanding of the mechanisms
involved. The basic need is methods to
study the effects of the environment on the
organism. These analytical methods
should provide a synthesis with predictive
value for assessing the response of a given
species to a new situation. The theoretical
basis for such an approach is not yet worked
out. A beginning can be made, however,
by describing the relationships between
environmental factors such as temperature,
salinity, and the concentration of
Reprinted from the Research Needs, National Technical Advisory Committee on Water
Quality Criteria, FWPCA, USDI.
BI.EN.2.7.69
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Fish, Other Aquatic Life, and Wildlife
respiratory gases and the ability of an
organism to succeed. Measuring the
performance of the active and standard
metabolic rates in key organisms with
multifactorial combinations of various
components in the environment would
provide a valuable base line for deter-
mining the effects of pollutants. Research
units should be organized to study the
environmental physiology of aquatic
organisms. Goals of the units would be
to determine metabolic requirements of
activity under a given set of environmental
conditions and to understand the mechanisms
which make such performance possible.
Parallel studies should also be made on
the effects of environmental changes on
the structure of the aquatic community
(kinds, distribution, and abundance of
species), food pathways, and energy flow.
It is essential that we learn how environ-
mental changes affect ecosystems, how
ecosystems react to toxicants, under what
conditions a new desirable ecosystem may
develop, and what types of stresses
initiate a chain of reactions which are
extremely difficult, if not impossible, to
stop.
A Selection of Test Organisms
The environmental criteria essential to
aquatic life resources are difficult to
determine. Extended, intensified, and
sophisticated research is required to
understand and manage the physical,
chemical, and biological environment
of fish, other aquatic life, and wildlife.
The identification of environments which
are (1) acutely harmful, (2) deleterious
under chronic exposure conditions, or
(3) safe under conditions of continuous
exposure are very involved problems
requiring research into the indefinite
future.
These problems are compounded by (1)
the large number of species involved,
(2) the wide range of climatic conditions
encountered in the United States and
its territories, (3) the inherent differ-
ences among marine, estuarine, and
freshwater environments, and (4) rapid
industrial progress with the evolution
of new contaminants, both real and
potential.
These problems are so numerous that
"short-cut" methods for finding
answers must be developed. A
desirable "shortcut" is limiting the
total number of species of organisms
to be studied, without creating serious
gaps in the knowledge required. This
should be possible because, in a given
faunistic region, the number of
abundant species which are important
are relatively few in comparison to the
total number in the area. It is rec-
ommended that test organisms be
selected from those species of major
economic and ecologic importance
which can be held and reared success-
fully in the laboratory.
Even with this reduction, the number
of species to be studied would be great
and perhaps beyond the resources
available for long-term studies. To
help alleviate this problem, it is
suggested that short-term sensitivity
studies be done to determine which
species and life stages are most
sensitive to each environmental
parameter or potential toxicant.
Subsequent long-term studies to
determine safe levels could be limited
to the most sensitive species and life
stages. The Committee believes that
if the most sensitive species and life
stages are protected, the entire biota
will in most instances be protected.
In some cases, it may be desirable to
associate these studies on a priority
basis with known wastes such as
sulfite waste liquor or pesticides.
All the different faunistic areas of the
Nation must be studied. These include
Alaska, Hawaii, the Commonwealth of
Puerto Rico, the Virgin Islands, Guam,
and American Samoa, as well as the
various river basins, and the principal
coastal zones.
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Fish, Other Aquatic Life, and Wildlife
Studies should be made to determine
(1) the most important species in a
particular biota, (2) methods for main-
taining these species and their various
stages in the laboratory so that they
may be available in numbers sufficient
for bioassays, and (3) the relative
sensitivity of the various test organisms
and life stages to the environmental
factors and toxicant under investigation.
By limiting sensitivity studies to those
species peculiar to a given faunistic
area, the total number under study
could be limited, but collectively all of
the nationally important species would
be represented. When such an arrange-
ment is operative, the species and life
stages that are the most sensitive to
each of the materials or wastes under
study can be determined in a routine
manner.
B Methodology
Even with the simplified outline
described above, the task of deter-
mining safe levels remains prohibitive
from the standpoint of time and cost
unless short-term evaluations can be
correlated with specific long-term
changes or effects. It is, therefore,
essential to devise and test several
biological parameters which may
furnish rapid indexes of environmental
requirements and safe concentrations
of toxicants.
In developing short-term methods, it
is recommended that the procedures
used in human toxicology be evaluated
for applicability. Tests should include
behavioral, physiological, biochemical,
enzymatic, endocrine, metabolic,
histological, histochemical, histo-
pathological, and various blood studies
when they appear necessary.
Current methods of bioassay indicate
the upper levels at which a potential
toxicant is intolerable. Short-term
bioassays as now developed and used,
do not indicate whether an environ-
mental factor is at an optimum level or
if it is deficient. It is recognized that
short-term bioassays are inadequate
for many purposes and that flow-through
and long-term bioassays are needed.
Many substances which act as
environmental poisons depress energy
utilization. Studies of energy utilization
should permit determining when sub-
stances are relatively innocuous as
well as when and to what degree they
slow metabolism. It is proposed,
therefore, that new approaches to
water quality be sought. By using the
energy concept, it may be possible to
express water quality criteria m a more
adequate fashion and research to
accomplish this goal should have
priority. Some of the suggested short-
term methods for determining
long-term effects may be effective in
reaching these goals.
Another gross type of index may relate
oxygen utilization and nitrogen ex-
cretion (the Og Ng ratio) to biochemical
imbalances and metabolic involvements.
The oxygen-nitrogen ratio may possibly
indicate the substrate being used for
energy although it may not distinguish
between fat and carbohydrates.
Combined with COg measurements,
the ratios may offer a reasonable
insight into the probable nature of sub-
strates utilized. Abnormal oxygen-
nitrogen ratios might result from
certain types of sublethal metabolic
anomalies that are caused by exposure
to some toxicants.
Due to the magnitude of research needs
for establishing meaningful water
quality requirements for fish and other
aquatic life, major emphasis will have
to be placed on laboratory bioassays
and on developing new and better
bioassay techniques to meet existing
problems. Increased efforts, there-
fore, must be expended to make
bioassays more realistically reflect
situations that occur in the field under
natural conditions. Properly conducted
experiments for comparing laboratory
results with field observations will
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Fish, Other Aquatic Life, and Wildlife
provide a sound scientific basis for
establishing correct criteria with
respect to temperature, oxygen, and
other environmental requirements.
Methods must be developed for detecting
subtle changes which indicate slow
deterioration in the aquatic environment.
These studies are essential if methods
are to be developed for field use,
because slow deterioration is the most
difficult effect to detect and prevent.
Subtle environmental conditions or low
concentrations of potential toxicants
must be studied. While these do not
cause death or any easily-observed
harmful effects, they will diminish
growth and productivity, decrease the
general well-being, or even in time
destroy the population or eliminate a
species. This is a problem similar to
the one faced in the protection of man
from the effects of air and water
pollution. Death is obvious, but the
gradual lowering of the general health,
well-being, productive capacity, and
slow deterioration of physical ability
are not so obvious. There must be
research to develop methods to detect,
measure, evaluate, and control the
subtle effects of pollution so that aquatic
life resources on which we depend for
food, recreation, and environmental
pleasures can be restored and main-
tained.
IH RESEARCH NEEDED
A Chemical Investigations
In the past, chemical analyses made in
connection with water pollution surveys
and investigations have not been
specifically directed toward toxicological
problems. Analyses were confined to
those materials for which analytical
methods had already been developed.
Usually, these tests were not sensitive
or accurate enough to detect and quan-
titate materials at the low concentrations
at which they are chronically toxic or
safe. Also, tests were not designed
to quantitate materials in their toxic
forms. For example, total cyanide is
measured as CN ion. This is not only
misleading but inappropriate in a
toxicological sense. Not all cyanides
are equally toxic and when they are
altered to and measured as CN ions,
the result is that more cyanide is
reported than actually occurs in one
or several toxic forms. Experimental
evidence indicates that it is the
undissociated HCN molecule which is
taken up by the organism. Thus,
methods must be developed to detect
and measure undissociated HCN in
mixtures of complex cyanides. This
is essential for the meaningful toxi-
cological evaluation of cyanide wastes.
The same is true for the H2S molecule,
the un-ionized NHg and the NH^OH
molecule. Research on analytical
methods for the development of micro-
analytical toxicological procedures are
essential for a sound water quality
program. Specific analytical methods
must be developed for all potential
toxicants so that they can be measured
in water and tissues at the very low
concentrations at which they are
chronically toxic. At present, only a
few analytical procedures are available
while several others have not yet been
refined to meet the speed and sensitivity
needed in toxicological work.
Research is needed in the field of bio-
chemistry for the detection and
quantitation of enzymes, metabolic
products, hormones and changes
resulting from exposure to toxic
materials. Special emphasis must be
placed on developing analytical methods
of the specificity, sensitivity, and
accuracy essential m toxicological
evaluations.
Chemicals or wastes that affect the
different functions or organs should
be evaluated, 1. e., materials or
families of products which affect
respiration, digestion, reproduction
and, on the molecular basis, the
enzymatic systems involved in these
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Fish, Other A quatic Life, and Wildlife
functions. The particular physico-
chemical changes tested should be those
induced by materials introduced in the
environment and known to produce
definite stresses on the organism.
These materials should be studied under
both laboratory and field conditions.
Furthermore, methods are needed to
determine the effect of two or more
biologically active materials that act
either in unison, antagonistically, or
synergistically. The causes and
mechanisms of synergism and antago-
nism should be examined. Knowledge
of the basic governing principles may
serve in forecasting consequences that
result from introducing specific products
or wastes.
Methods should be developed employing
radioactive tracers to aid in determining-
toxicant-altered metabolic pathways,
the metabolism of toxicants themselves,
and their turnover time or concentration
sites. Chemical and automated physico-
chemical techniques should be adapted
to complement ecological investigations.
B Environmental Requirements of
Aquatic Organisms
Information is needed on the essential
and optimal environmental requirements
of aquatic organisms. While some
work has been done on temperature
effects and oxygen requirements, it has
been limited to a few species that are
adaptable to laboratory conditions.
Precisely controlled and monitored
research must be carried out with the
important and most sensitive species
of each of the major classes of orga-
nisms to determine favorable and
allowable variations of environmental
factors.
Some work has been done on short-term,
acute lethal changes but the effects of
long-term exposures must be evaluated.
Data on the adaptive processes asso-
ciated with the sublethal changes are
essential in determining environmental
conditions favorable for the well-being
and the production of a crop of aquatic
organisms. Studies are needed on the
interactions of these environmental
factors and the levels at which various
combinations of these factors are most
favorable. Research needed on some
environmental factors will be listed and
discussed in turn.
1 Dissolved materials
Included under this heading are
those materials which are not
usually considered as potential
toxicants because they are only
toxic when present in abnormally
high concentrations. The most
common materials falling within
this category are the salts of the
earth metals. Studies should be
made to determine for aquatic
life the most favorable levels of
these materials alone and m
association with other materials,
as well as the most favorable
relative concentrations.
2 Turbidity, settleable and
suspended solids
Settleable and suspended solids
and turbidity are of great impor-
tance in the aquatic environment.
Any particle suspended m water
absorbs light and decreases
transparency. The effect of
attenuating light cannot, however,
be separated from physical,
chemical, and biological effects
of suspended materials on the
aquatic biota. Only the decrease
in the rate of photosynthesis can
readily be linked to increased
turbidity. However, sedentary
organisms, their eggs, and
larvae may be affected by the
settling out of excessive amounts
of silt, clay, detritus, and other
materials suspended in the water.
Plankton, expecially the diatoms,
may be covered with sediment and
lose their buoyancy, thus settleable
materials may effectively reduce
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Fish, Other Aquatic Life, and Wildlife
the plankton population by settling
out. The gills of fishes and bivalves
can be injured by sediments, there-
fore, the maximum concentrations
which are not harmful to important
species should be determined.
Research is needed on the following-
a The acute and chronic effects
of various concentrations of
presumably inert substances,
such as calcareous silt, sand,
kaolin and others can adversely
affect the respiration, feeding,
reproduction, growth and
behavior of important aquatic
species. Examples of these
species are bivalves, crabs,
shrimp, lobsters, fishes of
commercial and recreational
importance, and their food
organisms.
b Behavioral effects of silting
and sedimentation on the aquatic
biota under simulated natural
conditions where variables are
controlled.
c Physiological, chemical, and
biological effects of surface
phenomena associated with
suspended particles, their
affinities for different chemi-
cals, and their use as substrates
for microorganisms.
d The determination of the
qualitative and quantitative
penetration of different kinds
of light in relation to the kinds
and sizes of suspended
materials.
e The determination of the light
requirements of plants, both
quantitatively and qualitatively.
f Development of methods for
determining the physiological
effects on fishes and aquatic
invertebrates of long exposure
to high turbidities. Laboratory
studies are needed to deter-
mine tolerable levels of
turbidity or suspended
materials for different groups
of aquatic organisms such as
phytoplankton and zooplankton,
molluscs, aquatic insects
(especially the Plecoptera,
Emphemeroptera, Trichoptera,
and Diptera) and effects on
their productivity.
g Laboratory studies to deter-
mine the effects of turbidity on
migration of organisms and its
influence on flow-drift of
organisms.
The effects of settling materials
on the bottom have been studied
to some extent, but much more
information is needed on the
direct and indirect effects of
turbidity on species available
for food or recreation.
Laboratory and field studies
are needed to determine the
effects of sediments on aquatic
organisms and the amounts of
these materials which are
detrimental. Studies are needed
to indicate specific effects of
these materials and measures
or indexes of tolerable limits.
3 Color and transparency
Research is needed to determine
the long-term biological effects of
25, 50 and 100 units of color
derived from different sources on
the growth and reproduction of
phytoplankton, zooplankton, benthic
organisms, and submersed plants.
Information is needed on the extent
to which a particular color affects
the amount of light received at a
given depth. Studies should be
made on the biological consequences
of color independent of the direct
biochemical effects of the products
inducing the color.
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Fish, Other Aquatic Life, and Wildlife
4 Salinity
Emphasis should be placed on the
physico-chemical properties of
seawater. Research is needed to
determine the tolerable and favor-
able ranges of salinity for important
organisms and the variable toxicity
of pollutants at various levels of
salinity.
There is still much to be learned
about the physiology and behavior
of estuarine animals or amphibiotic
organisms migrating through
estuaries. How do they regulate
their internal environment under the
radical changes of salinity9 What
are the energy requirements under
different combinations of the varying
characteristics of estuaries and
how do the organisms meet these
requirements' How do pollutants
affect animal resistance when com-
bined with euryhaline migrations'
How can the animal survive the
added burdens offered by pollution9
5 pH
Some of the general effects of pH
changes on other environmental
requirements and on the toxicity of
materials and wastes are known,
but much remains to be learned
concerning the pH ranges which are
tolerable and favorable for a large
number of species. Data are needed
on the specific requirements of the
various organisms and the limits--
upper and lower--of pH which affect
them adversely or are lethal.
More study is needed on the effects
of pH on toxicity, oxygen require-
ments, tolerable carbon dioxide
levels, and the availability and
utilization of nutrients.
6 Temperature
Problems of temperature and
dissolved oxygen are interrelated
and should be considered as inter-
dependent variables. Research on
the following points is urgently
needed. Do daily fluctuations in
temperature have any biological
significance9 How rapid can the
change be in either direction9
Does it matter if the fluctuations
are out of phase with the natural
cycle9 What are the relative
effects of wide fluctuations above
a daily mean9 How much variation
can incubating eggs withstand9
Information is also needed on the
effects of seasonal fluctuations.
Are seasonal fluctuations essential9
How important are they in trigger-
ing spawning9 Are they required
for successful incubation of the
eggs9 Do they affect availability
of food organisms9 There is need
to know how much of a temperature
shock fish and other important
aquatic organisms can withstand
if swept into plumes of heated
effluents, or if fish, especially
amhibiotic forms, avoid areas of
unfavorably high temperature.
There is need to determine the
transition in temperature that
migrating fish can tolerate.
Another outstanding problem is that
of the selective favoring of species
by changes in temperatures and
thus altering or nullifying an
endemic balance of species. There
is need to know the levels of tem-
perature which are tolerable and
favorable for all important species
in the aquatic environment, marine,
estuarine, and freshwater. At
what temperatures will desired
species be replaced by undesirable
competitors9 As temperatures
increase, what are the trade-offs
relative to increased growth and
total productivity versus parasites,
disease, shifts in species com-
position, and overall mortality9
Information is required on the
levels of temperatures which are
tolerable and favorable for all
important species in the different
aquatic environments. Data are
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Fish, Other Aquatic Life, and Wildlife
required on the specific effects of
high, but sublethal temperatures,
the duration of elevated tempera-
tures which may become lethal,
those which are detrimental and
those which favor optimal develop-
ment. Studies should be carried
out to determine the ability of
organisms to withstand long periods
of high sublethal temperatures which
extend beyond the normal summer
period. Data are needed on the low
temperature requirements of
important organisms and the effects
of these low temperatures on the
development of sex products and the
completion of the normal life cycle.
Studies are needed to determine if
the acute or chronic effects of sub-
lethal, high temperatures become
more harmful when compounded with
stress, such as toxicants, starvation
or salinity changes. Information is
urgently needed on the temperature
requirements for spawning, egg,
and larval development of all species
important for recreational,
commercial, and forage purposes.
Analysis of the behavioral and
physiological mechanisms resulting
in a particular performance should
reveal some of the basic effects of
temperature. Thermal tolerance
diagrams are also needed for a
larger number of the economically
important freshwater and anadro-
mous fishes. Temperature-selection
curves and temperature-activity
curves are required for most if not
all important species.
7 Oxygen
While a considerable number of
studies have been carried out to
determine the minimum dissolved
oxygen concentrations required by
certain species, a great deal of
research is needed to determine
the dissolved oxygen concentrations
tolerable and favorable for all
important species. The effects of
low as well as high oxygen
concentrations should be
investigated. The effects of
various continued levels of
dissolved oxygen on growth rate,
as well as the effects of period-
ically induced variations in
oxygen content, and short-term
exposures to low or high concen-
trations of dissolved oxygen should
be determined. The metabolic
rates measured as oxygen
consumption by certain important
species under various conditions
of temperature and salinity of the
water should also be evaluated.
Tests should be conducted under
controlled conditions, in properly
designed and constructed metabolic
chambers. It is desirable that
each test be conducted for significant
periods of time to obtain accurate
levels of basal metabolism.
Carbon dioxide produced during the
test should be measured to obtain
the respiratory quotient values
(RQ). Effects of toxicants on
metabolic rates, oxygen require-
ments and RQ should be studied
using the important species for
which controlled measurements
have been previously obtained.
Studies are needed to determine
the oxygen requirements for the
fertilization and development of
eggs, development and growth of
larval and juvenile forms as well
as the effects of dissolved oxygen
on the activity and reproductive
cycle of the adults.
8 Carbon dioxide
Information is needed on the levels
of carbon dioxide which are
tolerated for short or extended
periods by various organisms and
levels which do not produce un-
favorable effects under conditions
of long-term exposure. Information
is needed on the photosynthetic
needs for carbon dioxide. Buffer
systems that are able to insure
desired productivity should be
developed. Buffering systems
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Fish, Other Aquatic Life, and Wildlife
essential to insure desired pro-
ductivity should be developed to
promote desired growth. Extensive
research is needed for determining
the concentration of nutrients which
produce undesirable blooms.
9 Nutrients and nuisance growths
Some of the factors contributing to
excessive growth are known,
however, concentrations and relative
proportions of fertilizing material
and other environmental factors
essential to the production of un-
desirable blooms are unknown.
Laboratory experiments can help
define the conditions which promote
these blooms in nature. In carrying
out such studies, experiments of
graded complexity can proceed from
controlled laboratory experiments
towards the sum of variables and
the near duplication of the natural
environment. Eventually,
experiments should be carried out
in controlled sections of natural
streams, or in artificial streams,
lakes, reservoirs, and estuaries
to duplicate natural conditions. In
this way, it would be possible to
study specific chemical and physical
factors as they affect particular
species living in a complex eco-
system, as well as the effects of
such factors on predatory relation-
ships. Questions to be answered
by research are, to name a few
a How are nutrient balances
affected by the addition of
given chemicals, under the
influence of various day lengths,
pH, temperatures, etc. 9
b How do shifts in nutrient balance
affect the abundance of diatoms,
blue-green algae, green algae
and rooted aquatics9
c How do these shifts affect the
abundance of species which
have high or low food values9
d What species are the pre-
ferred food for the important
organisms in the ecosystem9
e Is a diversified diet of many
species (diatoms and some
green algae species) a better
source of nutrition than a
single species source9
f As the human population and
its organic waste continue to
increase, how can nutrients
be balanced or treated to
promote desirable growths
rather than nuisance blooms9
g What are the best methods for
removing nutrients from water
and for the control of
eutrophi cation 9
h How can mineralized wastes
and sludges be used produc-
tively 9
l What kinds of bacteria and
fungi are important in
assimilating wastes in rivers
and what environmental factors
stimulate their development
and the effective conversion of
wastes to harmless materials9
j How can the heat discharged
in an effluent be used with
beneficial effects or with
minimized harmful effects9
k What are the degrees of inter-
dependence among species of
groups9 Studies on the effects
of limiting species diversity
could aid in this.
1 Which are the most important
organisms in the food web9
m Is energy more efficiently
transferred through a high or
low diversity of species within
a food web9
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Fish, Other Aquatic Life, and Wildlife
n How can our rivers be managed
so that the shallow water areas
support desirable aquatic
organisms9
C Toxic and Damaging Materials
1 General
Largely due to the activities of man,
many toxic materials are already
present in the aquatic environment.
The number is increasing con-
tinually due to the proliferation of
new materials and industrial
development. There already exists
a great backlog of materials for
which the acute toxicity to aquatic
organisms should be determined.
An even greater task is the deter-
mination of the long-term effects
of these materials. While short-
term bioassays have indicated the
acute toxicity of several wastes to
a few aquatic organisms, there is
little information on concentrations
of these materials which are
chronically harmless to aquatic
organisms of recreational, economic,
and forage importance. Safe levels
of these materials must be deter-
mined for all important sensitive
species if the biota is to be pro-
tected. Information is lacking also
on the possible antagonistic or
synergistic effects of combinations
of these materials on selected
species.
2 Tastes and odors
There are a number of materials
which may, at sublethal concen-
trations, taint fish flesh, or
produce odors and tastes which
are undesirable. Studies are needed
to determine those materials and
organisms that produce tastes and
odors in water supplies, as well as
methods for their elimination on
control. Studies should be carried
out to determine materials that
taint flesh of aquatic organisms,
the length of time required for
tainting, the maximum concen-
trations of these tainting substances
which do not produce tastes or
odors, and the time needed to
render the tainted organisms for
use.
3 Oils
Studies should be conducted at sites
of oil pollution to determine
a Chemical composition of oil
slicks at their point of origin
and at increasing distances
from it and the composition of
oil sludges and their change in
relation to time and distance
from the spillage source.
b Accumulation of water soluble
toxic compounds in oil
polluted waters, identification
of them, and determination of
their toxicity.
c Rate of degradation of oil under
controlled laboratory conditions
and in the natural environment
with special emphasis on the
effects of oxidation on oil
toxicity.
d Gas formation in oil sludges,
its identification and the
carrying capacity of gas
bubbles m moving or trans-
porting oil to the surface to
form new oil slicks.
e Interaction of various water
soluble fractions of oil with
the organic compounds
naturally present in the water.
Studies should also be made to
determine the following
a The survival of adults, larvae,
and eggs of selected species in
different concentrations of
various grades of crude oil,
fuel oil and kerosene.
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Fish, Other Aquatic Life, and Wildlife
b Effects of oil accumulation on
the gas exchange apparatus of
gill-breathing organisms.
Aquatic insects, oysters, soft-
shell clams, bay scallops,
flounders, etc. are of interest
here.
The following problems should be
investigated in the coming years.
a Anatomical injuries to fishes
and other aquatic animals
resulting from explosives used
for oil exploration and other
purposes.
b Effects of explosions on
behavior of individual animals,
school formation, and stock
migration.
c Effects of vibrations on
behavior of individual fish and
other animals and on populations
of important species.
d Effects of vibrations on spawning,
fertilization of eggs, and egg
development.
e Effects of explosions and
vibrations on plankton.
4 Highly toxic wastes
Most of the studies on the toxicity
of various materials to aquatic
organisms have been short-term
bioassays with pure compounds.
While a few studies on complex
wastes have been made, they have
been limited due to the nature of the
tests and the inability to secure
large amounts of these wastes.
Recently, a number of long-term
studies have been made where low
concentrations of toxicants have
been maintained over the entire life
cycle of a test organism. These
relatively few observations have
indicated that the application factor
to be applied to TL values to
denote safe concentrations during
continuous exposure may range
from 1/7 to 1/500 of the 96-hour
TL^. Application factors
recommended by the National
Technical Advisory Subcommittee
on Water Quality Requirements for
Fish, Other Aquatic Life and
Wildlife are 1/10, 1/20, and 1/100
of the 96-hour TL^. It was
realized that in certain instances
these factors would be either over-
protective or possibly inadequate.
Newly-determined application
factors should replace these
interim values as rapidly as
scientific observations are com-
pleted. Research to determine
application factors for the most
important wastes is imperative.
The following research projects
involving studies on highly toxic
wastes, should be given priority for
adequate protection of aquatic life.
a Development of methods for
raising all life stages of test
organisms in order to com-
plete essential laboratory
bioassay studies.
b Short-term bioassay studies to
identify the most sensitive
species and the most sensitive
life stage to selected materials
and wastes.
c Long-term bioassay studies
with those species and life
stages which are the most
sensitive to a particular waste
to determine the following
1) The consequential effects
of non-lethal damage at
any stage on successive
stages in the life history.
2) The effects on all life
stages of sublethal ex-
posures during the most
sensitive stage.
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Fish, Other Aquatic Life, and Wildlife
3) Comparative studies of the
effects of exposure during
the most sensitive stages
and of exposures during
the entire life-cycle.
d Determination of the normal
range of natural environmental
fluctuations on the toxicity of
various materials.
e Associated with investigation of
the long-term effects of various
toxicants, the following studies
should be made.
1) Determination of the mode
action, of toxicants, their
metabolites, and detoxifi-
cation mechanisms.
2) Determination of the inter-
action of two or more
toxicants, synergistic and
antagonistic effects, and
those materials with
similar and different
modes of action.
f Studies should be initiated to
develop short-term methods
for determining long-term
effects. These studies should
include the following
1) Behavioral studies to detect
any change that may render
the organism unable to
cope with the natural
environment or make it
susceptible to predation or
disease.
2) Physiological studies to
detect any adverse functional
effects of exposure to sub-
lethal concentrations of
toxicants.
3) Investigations of effects on
enzyme or endocrine gland
functions.
4) Metabolic studies
evaluating changes due
to the action of toxic
materials.
5) Blood studies for the
development of methods
that may indicate subtle
sublethal effects.
6) Histological, histochemical,
and histopathological
studies to detect changes
due to sublethal concen-
trations of toxicants.
7) Use and adoption of
methods developed by
medical science for the
detection of toxicants and
the evaluation of their
effects.
g Bioassay experiments should
be designed to permit adequate
statistical analysis of the
results. Reliable formulae
for the computation of appli-
cation factors could thus be
derived.
h Bioassay methods are needed
for arthropods, molluscs,
worms, phytoplankton, zoo-
plankton, and bacteria.
Studies are needed for
improvement and standardi-
zation of bioassay procedures,
for the utilization, development,
adoption, and improvement of
bioassay instrumentation, for
the monitoring and control of
water quality and for the
detection and the measurement
of pollutants.
l Research is needed to deter-
mine the toxicity of degradable
toxic products and to determine
the mechanism and rate of
their degradation.
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Fish, Other Aquatic Life, and Wildlife
5 Radioactive wastes
Research is needed for determining
the following,
a Behavior and fate of radio-
nuclides in the aquatic
environment and their con-
centration by aquatic food
organisms.
b The influence of the physico-
chemical state of the
radionuclides and the
characteristics of the dynamic
equilibrium within the eco-
system on the availability of
the radionuclides to the biota.
c The effects of environmental
conditions upon the biological
concentration of radionuclides
and their passage through food
chains.
d The damaging effects of the
continuous exposure of sensitive
stages of important aquatic
organisms to radionuclides.
D Ecological Field Investigations
Although laboratory experiments can
provide valuable information on how an
organism reacts under controlled con-
ditions, tney do not necessarily reveal
how an organism will react under the
stresses found within the complex
ecosystems of natural waters. There
are many instances where the predictive
values of such laboratory experiments
are poor This is probably due in great
measure to the largely artificial environ-
ment in which they are carried out
Therefore, some of these experiments
should be extended to areas duplicating
natural environments, such as pilot
areas or areas in sections of natural
waters which can be monitored or con-
trolled for the purposes of these studies
Ecological field studies are needed to
evaluate laboratory findings for the
favorable ranges of such environmental
factors as temperature, dissolved
oxygen, carbon dioxide, dissolved
solids, suspended solids, settleable
solids, turbidity, color, light require-
ments, currents, salinity, and pH.
1 Methods
Available field methods are not
adequate to meet the problems
which we now face in the detection
and measurement of various
environmental factors and their
effects on the aquatic biota.
There is a need to develop
facilities, equipment, and methods
for field testing so that necessary
studies can be carried out under
natural conditions for determining
harmful or favorable effects.
Better sampling methods are
needed for both grab samples and
continuous monitoring. More
accurate methods for compositing
samples and taking samples from
composites should be developed.
Better instrumentation is needed
for continuous recording of
environmental factors. Better
methods or models are needed for
relating the conditions in streams,
lakes, reservoirs, estuaries, or
coastal areas to various kinds of
land use and to various kinds and
amounts of effluents being dis-
charged into these waters. To
improve the evaluation of sublethal
or obscure effects of potential
toxicants, more precise field
methods are needed for determining
the effects of pollution on the
ecosystem and for measuring
standing crops and productivity at
the various trophic levels.
Autopsy techniques should be
developed for determining the
cause of kills, the concentrations
of the toxicants or their metabolites
in the organism, and the relation-
ship of these compounds to lethal
or sublethal effects.
2 Environmental requirements
a Temperature Field studies
on the effects of heat are
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Fish, Other Aquatic Life, and Wildlife
necessary under pilot plant
conditions or m the environment
where conditions are the same
or as nearly approaching
natural conditions as possible.
Ecological observations should
be conducted in the areas
affected by discharges of heated
water from power plants.
Experiments conducted in tanks,
troughs, or raceways under
controlled conditions should.be
carried out to determine the
survival of species studied.
These studies should include
effects of temperature and its
variations on the competition
of the different species, the
effects of extremes in tem-
perature and temperature
variations on the movement of
aquatic organisms, their
spawning, egg development and
survival, development of
larvae, the presence of growth
anomalies, and the production
of desired species.
b Dissolved oxygen Field studies
are needed to determine
oxygen concentrations which
are lethal, or unfavorable, and
those which are favorable under
natural conditions where there
is competition for food and
space, and where the other
stresses found in a natural
environment are present.
Such studies are needed for
fresh, estuarine, and marine
waters. Much can be learned
by field ecological studies of
the occurrence and well being
of populations of important
species in waters with normal
as well as different oxygen
concentrations and different
daily and seasonal variations.
c Toxic materials Special pilot
study areas, controlled natural
areas, or natural areas must
be used for the field evaluation
of laboratory findings on
toxicity. Natural areas may
also eventually be used for
determining the effects of a
toxic material or waste on the
entire biota of a water.
The direct translation of
laboratory data to the field
situation is very difficult and
often unsatisfactory. Under
natural conditions, there may
be a rapid reduction in the
concentration of a toxicant by
one or more of the following,
precipitation, adsorption on
soils and bottom materials;
chemical decomposition,
reactions with other sub-
stances in the water,
absorption by microscopic
organisms, removal by
organisms and biochemical
degradation. Accumulation
of these materials in the food
chain and ingestion of food
organisms bearing relatively
high concentrations of these
materials may increase the
exposure to the higher
organisms.
Laboratory findings on the
safe levels of potential toxi-
cants and environmental
requirements must be field
tested under conditions where
the organisms in question are
exposed to all the stresses
occurring in the natural
environment. Such tests
should be carried out before
water quality requirements
are recommended. When
developed, tested, and
evaluated, field studies can be
used for simultaneously testing
all the species in the biota
under natural conditions. In
such studies, biological
magnification, storage,
passage through the food
chain, accumulation in bottom
materials, competition for
food, cover and living space.
-------
Fish, Other Aquatic Life, and Wildlife
the effects of disease parasites
and predators, synergism,
antagonism, and the interaction
of materials and all other
complicating factors present
in the natural environment are
taken into consideration. When
effective studies can be made
in streams, lakes, reservoirs,
estuaries, and coastal areas,
the determination of safe levels
of potential toxicants in the
environment will be made in
much less time. Such field
studies will require develop-
ment of improved methods for
estimating the biota and
seasonal changes in the biota,
as well as development of
analytical methods for con-
tinually monitoring the actual
consideration of potential
toxicants in the areas under
test. Where known pollutants
occur and fishery statistics
are available, field studies
properly designed and carried
out can be the most convincing
means of determining effects
of such pollutants. A great
deal can be learned by eval-
uating the effects of nature's
bioassay, i.e., evaluating the
biota that has developed and
maintained itself under certain
environmental conditions.
Observations of populations
and population dynamics, in
different areas having different
levels of wastes, can be very
informative in indicating
relative sensitivity and the
overall effects upon the biota
of the additions of these wastes.
should be expanded to determine
1 Distribution of pesticides, heavy
metals, and trace elements in the
wildlife food chain and the tissues
of various species, expecially
migratory birds.
2 Analytical methods to differentiate
between the compounds present in
the environment versus the
absolute amount of compounds
available for absorption by plants
and animals, e. g. Hg and Se.
3 The plants and animals that con-
centrate specific chemicals to
levels capable of intoxicating other
organisms in the food chain.
Elements of toxic significance are
antimony, arsenic, beryllium,
bismuth, cadmium, chromium,
lead, lithium, mercury, nickel,
selenium, silver, tillurium, and
thallium.
4 In addition, studies should be made
on the effects of environmental
nutrient elements present in
excessive amounts or m deleterious
combinations. Among these are
barium, boron, calcium, cobalt,
copper, iron, manganese,
magnesium, molybdenum,
potassium, sodium, and zinc.
5 New or better analytical methods
for identification and quantitation
of nutrients, toxicants, and trace
elements, their movement through
the food chain, their biological
magnification, and where, when,
and in what form they accumulate.
Ill WILDLIFE
A The needs for wildlife are largely met
if the requirements for aquatic life are
satisfied. However, additional research
is needed for the determination of
special requirements essential to wild-
life. The following research studies
IV SUMMARY
The objective of this proposed research is
to determine the water quality requirements
of aquatic life and wildlife. To accomplish
this objective there must be an effective,
efficient, adequately supported, and
continuing national research program.
This program must be planned and carried
-------
Fish, Other Aquatic Life, and Wildlife
out so that adequate data are secured to
establish (1) maximum or minimum
environmental conditions and concentration
of toxicants which shall not be exceeded,
(2) environmental conditions and concen-
tration of toxicants which can be tolerated
for short periods without significant harm,
(3) the range of environmental conditions
and concentrations of dissolved materials
which are not harmful and those which are
favorable and required for all important
species.
A Research Approaches, Methods and
Procedures
The effects of the environment
on the total biota and
individual organisms to pro-
vide a synthesis with
predictive values for indicating
the responses of a species or
biota to a given change in the
environment.
The active and standard
metabolic rates under multi-
factorial combinations of
various environmental
components in the presence of
pollutants.
Research must be conducted to deter-
mine new or better methods for the
following purposes.
1 To devise or modify and develop
equipment and methods for the
collection, isolation, and production
of all life stages of important
aquatic organisms so they are
available at all times in sufficient
numbers for bioassay studies.
2 To devise or modify and develop
effective bioassay methods and
other procedures for determining
environmental conditions and con-
centrations of potential toxicants
which should not be exceeded,
which can be tolerated for short
periods and which are favorable
and essential for the survival,
growth, reproduction, and general
well being of all important groups
of aquatic life and wildlife.
3 To develop facilities, equipment
and ecological, analytical,
physiological, toxicological, and
other methods for the field and
laboratory determination of the
effects on the biota of alterations
in environmental conditions and the
presence of various concentrations
of potential toxicants.
4 To develop special equipment or
methods for determining or
developing-
c New and more specific
methods for detecting sublethal
effects of toxicants such as the
oxygen-nitrogen ratio and
bioenergetics.
d Methods for sampling and
monitoring, instrumentation
for automatic continuous
sampling, more precise field
methods for determining effects
of pollution, and determination
of standing crops, productivity,
and population dynamics.
e Autopsy techniques
B Chemical Investigations
Research is needed for.
1 The development of microanalytical
and other toxicological methods for
the detection and quantitation of
toxic materials in water and tissues
at the extremely low concentrations
at which they are either toxic or
safe under conditions of continuous
exposure.
2 The development of analytical
methods of the specificity,
sensitivity, and accuracy
essential for toxicological studies.
3 Developing methods for using
radioactive tracers to aid in
determining metabolic pathways
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Fish, Other Aquatic Life, and Wildlife
of toxicants, their breakdown
products, and their turnover time
or concentration in tissues.
C Environmental Requirements of Aquatic
Organisms
Research should be carried out to
determine the environmental require-
ments of all stages of the life history
of important sensitive species to
indicate lethal levels or sublethal but
harmful levels of environmental factors,
and the ranges of these factors that are
favorable and promote the production
of a desirable crop. Research for these
purposes will include the following
studies.
1 Temperature and dissolved oxygen
Laboratory and field research should
be carried out to determine the
follow ing
a What is the significance of daily
and seasonal fluctuations in
temperature and dissolved
oxygen9
b How rapid may changes be
without exerting an adverse
effect'
c Does it matter if the fluctuations
are out of phase with the
natural cycle9
d What are the effects of wide
fluctuations above or below
the daily mean and how much
can temperatures be raised
and dissolved oxygen concen-
trations lowered from natural
conditions without harming the
biota or causing a change in its
makeup9
e At what temperatures and
dissolved oxygen levels will
desired species be replaced
by undesirable competitors9
f What are the trade-offs
relative to increased growth
and total productivity versus
parasites, disease, and
mortality of different life
stages9
g What levels of temperature
and oxygen are harmful,
tolerable, and favorable for
all the important species at
all life stages, including egg,
nymph, larvae, fry, etc. 9
h What are the effects of changes
in daily and seasonal temper-
atures on migration, scope of
activity, and the toxicity of
waste materials9
1 What are the maximum tem-
perature and minimum oxygen
levels that can be tolerated for
short periods without harm9
j What are the allowable overall
increases in temperature at
different seasons that do not
produce undesirable effects
on the biota9
k What are the allowable and
favorable levels of temperature
and oxygen for all important
species9
1 What are the requirements for
low temperatures in the life
cycle of important organisms9
m What are the effects of stress
due to pollutants on the
temperatures which are lethal,
tolerable, or favorable9
n What are the beneficial effects
of the addition of heat9
2 Nutrients and eutrophication
a What are the nutrient require-
ments of desired species and
the organisms in their food
web9
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Fish, Other Aquatic Life, and Wildlife
b How much can nutrient con-
ditions be changed from the
normal without detrimental
effects'
c What are the amounts of nutrient
or fertilizing materials which
may be desirable from the
standpoint of productivity?
d How do changes in nutrient
balance, due to additions of
wastes or other materials,
affect the abundance of differ-
ent groups or species of
organisms and the eutrophication
of a water'
D Other Environmental Factors
Dissolved materials Determination of
high or low concentrations which can be 2
tolerated for short periods and the
favorable range of concentrations or
levels for the following, dissolved
solids, suspended solids, settleable
solids, turbidity, color, salinity, pH,
and carbon dioxide.
E Toxic Wastes and Materials
The following research should be
conducted in connection with the
toxicological program.
1 Long-term studies with the most 3
sensitive species and life stages
of important organisms should be
conducted. They would serve to
determine the maximum concen-
tration of potential toxicants which,
under the extremes of allowable
environmental conditions, are not
harmful with long-term or
continuous exposure as indicated
by survival, growth, activity,
reproduction, and the general well-
being. Studies for determining
long-term effects should include
the following:
a Behavioral studies to detect
any changes that may render
the organism less capable to
cope with the natural environ-
ment or make it more
susceptible to predation or
disease.
b Physiological studies to detect
any adverse functional effects
of exposure to sublethal con-
centrations of toxicants.
c Investigations of effects on
enzymes or endocrine gland
functions.
d Metabolic studies to evaluate
changes due to the action of
toxic materials.
e Histological, histochemical,
and pathological studies.
Devise, develop, and test short-
term methods for determining
long-term effects of toxic
materials on aquatic organisms
through the use of metabolic,
physiological, biochemical,
behavioral, histological, histo-
chemical, histopathological,
enzymatic, hormonal, stress,
energy transfer and balance, and
other tests carried on in con-
junction with long-term laboratory
tests.
Studies to determine the following.
a The effects of non-lethal
damage during any develop-
mental stage on successive
stages.
b The effects on all life stages
of sublethal exposure during
the most sensitive stage.
c Comparative studies of the
effects of exposure during the
most sensitive stages and the
complete life cycle exposures.
d The mode of action of
toxicants and mechanisms
of detoxification.
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Fish, Other Aquatic Life, and Wildlife
e The interaction of two or more
toxicants, synergistic and
antagonistic effects, and
different modes of action.
4 Determination of application factors
for use with TLgg values to deter-
mine safe discharge rates for
industrial wastes.
F Radioactive Waste
Studies should be conducted to determine
allowable levels of radioactivity in the
environment which are harmful to the
aquatic biota or produce concentrations
in the bodies of aquatic organisms
which render them unsuitable for food.
G Tastes and Odors
Research to determine- (1) materials
and organisms responsible for taste
and odors in water supplies and the
flesh of edible organisms, (2) the
maximum concentrations of these
materials which do not produce objection-
able tastes or odors and methods for
their control.
and where, when, and in what form
they accumulate. Other needed
investigations include'
1 Studies on the sources, disposal,
treatment and elimination of oils
from the habitats of waterfowl.
2 Studies on the effects of pollutants
or changes in water quality on
waterfowl and wildlife food
organisms.
3 Determination of concentration of
herbicides (used for controlling
nuisance plants) which do not
produce long-term pathological
effects or tainting.
4 Determination of the effects of
chemical mosquito control on
waterfowl and other organisms
in the biota.
H Wildlife Requirements
Research to determine new or better
analytical methods for identification
and quantitation of nutrients, toxicants,
and trace elements. Also, we need to
know about their movement through the
food chain, their biological magnification,
-------
SECTION G
SOME CURRENT POLLUTION PROBLEMS
Water usage generally is followed by return of the used water to the source
from which it was taken, or to some other body of water. Alteration of the
characteristics of water typically results from usage. Wherever these
alterations interfere with subsequent use of the water, they are regarded as
pollution.
In this Section, a selection of outlines identifies typical changes in water as
a result of current uses, and describes the general nature of the effects of
such use on the water.
Land use, eutrophication of lakes and streams; thermal pollution; the
phenomena of biological magnification (including mutagens, teratogens, and
carcinogens), oil spills; siltation; and non-point sources are all current problems
of environmental management greatly significant to man.
Contents of Section G
Outline No.
Effects of Pollution on Aquatic Life
31
The Effects of Organisms on Pollution and the Environment
31
The Effects of Pollution on Lakes
32
Water Temperature and Water Quality
33
Effects and Control of Oil Pollution
34
Biotic Effects of Solids
35
Global Deterioration and Our Ecological Crisis
-------
EFFECTS OF POLLUTION ON AQUATIC LIFE
I INTRODUCTION
A The effluent from any given industrial
plant may be combined with municipal
sewage, and/or wastes from other
industries. This may occur in the
sewerage system or in a natural body
of water.
1 Toxic wastes may inhibit biota of
treatment plant as well as life in
receiving stream.
2 Organic wastes may simply increase
sewage-type load on plant and stream.
3 The above effects may be reinforced
or neutralized by a complex of
industrial wastes.
B The general overall character of a body
of water may be subtly changed over a
period of time.
II The principle of limiting factors (see
Figure 1) deals with the response of
organisms to various factors in the
environment.
A Liebig's Law of the Minimum (Figure 1)
states that the distribution of a species
may be limited by one or more essential
environmental factors which occur in
minimal quantities.
B Shelford on the other hand pointed out in
his Law of Tolerance (Figure 1) that there
are also maximum values of most
environmental factors which can be
tolerated. In between these two extremes
there are ranges which may be called
"optimum" for factors useful to the
organism. Purely deleterious factors
on the other hand have a maximum tol-
erable value, but no optimum. The
range between the maximum concentration
(greater than zero) which kills no orga-
nism and the minimum concentration
which kills all organisms is known as
the "critical range."
C These principles apply to all aquatic life
whether in a stream, lake, estuary, or
treatment plant. They are the basis for
the control or regulation of biological
conditions.
in INDIRECT TOXICITY: MODIFICATIONS
OF THE ENVIRONMENT WHICH AFFECT
AQUATIC LIFE
A Deposition of inert precipitates and silt
tends to smother bottom organisms.
Contributing materials include silt or
sand from erosion due to poor agricul-
tural practices, rock flour or tailings
from mining or quarry operations, mica,
coal washings, sawdust and debris from
lumbering, insoluble precipitates or
complexes from chemical industries.
1 Vulnerable organisms include
important fish foods such as insect
larvae and snails, also fish eggs,
bottom-living algae such as diatoms,
and many others.
2 Physical injury to delicate membranes
of eyes, and gills may also result.
3 Inert suspended materials and dyes
reduce light penetration, suppress
photosynthesis and hence biological
productivity. They also prevent game
fish and other predators from seeing
their prey, thereby reducing the
efficiency of food utilization.
The word "stream" should be interpreted in
most cases to mean "river, " "lake, "
"estuary, " etc., as applicable.
BI.BIO.Ilk.5.71
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Effects of Pollution on Aquatic Life
W
V
rH
o
rH
UD
a>
Liebig's Law
Shelfords Law
Optimum
Range
Extinction
Extinction
Low
Magnitude of Factor
High
Figure 1. EFFECTS OF ENVIRONMENTAL FACTORS
Wastes of significant heat content may
change the "climate" of a body of water.
Temperature may be higher or lower
than normal.
1 Abnormally low summer temperatures
may prevent the reproduction of some
of the typical inhabitants, on the other
hand colder water forms such as trout
may be enabled to survive.
2 Abnormally high winter temperatures
may encourage a rapid development
of some species, thus for example,
causing an early emergence of some
insect followed by its death from normal
winter temperatures. Southern forms
may also invade more northerly waters,
sometimes leading to a year-round
nuisance from flying swarms of adults
such as caddis flies.
Each species however, has some
maximum temperature above which
it cannot survive (Figure 1).
a Quick temperature changes are
fatal at much lower values.
b Lack of oxygen due to low solubility
at high temperatures or from an
increase in the rate of the BOD,
also contributes greatly to high
temperature mortalities.
C Oxygen-consuming wastes kill by depleting
the free dissolved oxygen resources.
1 Amount present, rather than percentage
of saturation, is usually more significant.
2 Minimum amounts rather than averages
are most critical.
3 Artificially produced high temperatures,
often lack dependability. It is dis-
couraging for organisms to speftd six
days in summer temperatures in
January, only to freeze to death over
the weekend because no one warned
them that the plant would shut down'
4 Excessively high summer temperatures,
even for a few hours once a year,
probably represent the greatest tem-
perature danger. Some species of fish
can adjust to temperatures approaching
100° F, if the change takes place slowly.
a Any species can survive something
less than the optimum concentration
of DO for a limited period of time.
There is, however, some concen-
tration for any given temperature,
which Will eventually result in the
death of that species. Let us call
this the "critical" 'DO.
b As the DO falls below the critical
concentration, survival time
eventually drops to zero.
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Effects of Pollution on Aquatic Life
c The absolute values of these thresh"
olds vary with the species and other
factors. Five mg per liter is
often listed as a minimum permissible
value to maintain a well~rounded
healthy population of fishes on a year-
round basis.
3 Low oxygen tensions may also increase
the toxicity of certain chemicals.
D pH
1 "pH" is a logarithmic expression of the
hydrogen ion concentration in a solution.
Hydrogen ions (H+) in certain concen-
trations are toxic to aquatic life
(as are also hydroxyl ions OH").
Aquatic life in relative abundance and
variety can be found in waters ranging
from approximately pH 5 to 9. Thriving
communities including algae, insects,
and fish have been studied in waters
with a pH of at least 11.
2 Many species of aquatic organisms can
adjust to pH values over a wide range.
Sudden change of any kind however can
be fatal.
3 Most metals and other toxic substances
m dilute solution tend to become less
toxic at high pH values. A notable
exception is ammonia.
IV POLLUTION WHICH RENDERS FISH OR
SHELLFISH UNUSABLE OR INTERFERES
WITH THEIR CAPTURE
A Radioactivity at levels currently found in
our waters has not been observed to
adversely affect aquatic life itself. It may
. however, be taken up with food materials
and render fish or fishery products unusable.
1 Radioactive nuclides (forms of chemicals)
are taken up by the plants (predominantly
algae) in the processes of photosynthesis
and other types of protoplasmic syn-
thesis. There is no selection between
nuclides on the basis of radioactivity.
2 As the chemicals originally assimi-
lated by the algae are "eaten up the
food chain" (from algae to inverte-
brates to small fish to large fish to
man and other predators) their
radioactivity moves along with them.
3 Thus radioactivity is acquired by
fishes essentially through food, and
scarcely at all by direct assimilation
or absorption.
4 Plankton-feeding organisms such as
herrings, oysters, and clams acquire
radioactivity directly from the algae
on which they feed. Since they con-
centrate this food from large volumes
of water, they may be much "hotter"
than the surrounding water itself.
B Fish may be repelled or driven out of an
area by obnoxious chemicals. This may
simply result in their scarcity or absence
from a given locale, or it may prevent
their swimming up a river to spawn. In
this case the species would soon disappear
and be lost to the community.
C Color, odor, oil, floating scum, bacterial
slimes, and other such materials tend to
discourage sport fishing and interfere
with gear used by commercial fisheries.
D Sublethal concentrations of chemicals
such as phenol, benzene.oil, 2-4-D, etc.,
may impart an unpleasant taste to fish
flesh, even when present in very dilute
concentrations. This is nearly as
detrimental to the fisheries as a complete
kill, and of course applies to shellfish
as well as fin fish.
E Minamata disease was first described
from Minamata, Japan, as a disorder
resulting from eating various seafoods
taken from Minamata Bay, Kyushu,
Japan. The disease results from
industrial toxicity, m this case an organic
mercury compound, transmitted to a wide
variety of local marine seafood species.
These organisms are not known to be
affected, but acting as "transvectors, 11
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Effects of Pollution on Aquatic Life
pass the toxin along to predators or human
consumers. Over 30% mortality occurred
in Minamata among people eating local
seafood.
Acute toxicity can be evaluated by
means of the toxicity bioassay
technique and various modifications
(Figure 2).
It may be important to note that a fish
kill was recently reported from a TVA
lake m this country resulting from
mercury leaching from corroded 50
gallon drums used as floats by
marinas.
Bird and fish kills have recently
occurred in Swedish lakes resulting
from mercury compounds from pulp
mills. Levels in pike exceed the WHO
standards for human consumption and
the local population has been advised
not to eat pike more than once a week!
Cases of high mercury levels are increasing.
V DIRECT TOXICITY:
ORGANISM ITSELF
AFFECTS THE
Fish kills are often the result of direct
toxicity. If this is sufficiently potent to kill
at once, or within a few days, it is called
acute, and is often observed as a "fish kill.'
100
t 50
3
U]
-M
c
-------
Effects of Pollution on Aquatic Life
6 The result may be a slow and subtle
alteration of the characteristics of a
stream over an extended period of time.
C The specific physiological mechanisms
involved are infinite in variety and but
little known. Included are such processes
as enzyme inhibition as in the case with
some of the pesticides, and over stimu-
lation of mucous membranes of the gills,
leading to death by suffocation.
D There are a number of excellent diagnostic
techniques for the examination of dying
fish, these include pathogenic bacteria,
parasites, some metalp, and certain
pesticides. None are routine but require
specific handling and preservation tech"
niques •
E A recently developed procedure for
protecting aquatic life from deleterious
substances is biomonitoring. This is
the continuous monitoring or surveillance
of an effluent for toxicity by means of a
system for exposing living organisms
(such as fish or invertebrates) to a con-
tinuously flowing stream in a dilution just
below the known danger point. Should the
toxicity of the substance increase the test
organisms respond in some recognizable
manner, thus giving warning that correc-
tive measures need to be initiated.
VI MECHANISMS OF POLLUTION
TOLERANCE AND SENSITIVITY
The fact that some organisms are more
resistant to pollution than others needs no
emphasis. The matter of "why9" and "how''"
on the other hand, is quite another question.
In some cases the answer is obvious, in
others not. In general we can say that the
adaptations of certain species enable them
to resist certain types of natural conditions
such as organic deposits or sand bars. When
man artificially creates conditions such as
sludge banks, or sand bars, organisms which
can tolerate such conditions move in, survive,
and often thrive. Other forms are
eliminated.
A Organic pollution is essentially non-toxic.
Its typical result as noted above is
oxygen depletion, physical turbidity, and
smothering blankets of sediment or sludge.
B Devices and mechanisms for living in
oxygen-poor or oxygen-free water include
the following
1 Obtaining oxygen from the air by means
of periodic trips or access to the
surface.
a The snorkel tube of the rat-tailed
maggot.
b Periodic trips of the mosquito
larvae and Corixidae or water
boatmen. The mosquito takes air
directly into its respiratory system,
the water boatman into a trap or
space beneath the wing covers, as
well as into a layer of air held by
fine hairs or "pile" all over its body.
c Behavior of the air breathing snails
such as Physa which have an internal
lung cavity.
d Insects which tap into air tubes in
aquatic plants.
e Fishes which gulp air at surface or
breathe surface water.
2 Special devices and behavior for
respiration of water
a Hind intestine respiratory
structures of dragonfly larvae
permits respiration in silt-laden
water.
b Movement of gill covers and
similar structures in isopods and
certain msect groups maintains a
current of water over respiratory
organs.
c Body movements of chironomid
larvae create water current m
tibe. Sludge worms
-------
Effects of Pollution on Aquatic Life
and other annelids also create water
movement by means of sinuous body
movements.
3 Physiological and behavioral adaptations
to endure low oxygen tensions.
a Forms possessing accessory
respiratory pigments such as
hemoglobin might be expected to
be able to be able to extract the
last vestige of dissolved oxygen from
the water. Two groups famous for
resisting low DO do have hemoglobin:
the larvae of certain Chironomid
midge flies, and small annelid worms
such as sludge worms, (it should be
noted however, that the hemoglobin
in each case is simply dissolved in
the blood plasma, rather than being
concentrated in special corpuscles
as is the case in the more efficient
vertebrate system.)
b The mere possession of hemoglobin
however, does not seem to assure
tolerance of low DO (Walshe '47).
Larvae of the midge Tanvtarsus spp.
have hemoglobin, but will not
tolerate oxygen~poor waters.
Hemoglobin "bearing Chironomus
bathopilus is moderately tolerant,
and Chironomus olumosus is highly
tolerant, however.
c During periods of low DO, ChironomuB
plumosus apparently respires carbo"
hydrates as usual, but excretes
excess lactic acid instead of accu-
mulating it.
d Various species of Daphnia (micro-
crustacea) have been shown to
accumulate hemoglobin in oxygen-
poor waters but not in oxygen-rich
waters (Fox '47). No clear adaptive
significance has yet been proven
however.
C Advantageous Feeding Habits
All highly organic pollution tolerant orga-
nisms are scavengers, and hence find an
abundance of food. Most are relatively
defenseless and hence have normally high
reproductive rate. The result in a
polluted situation is thus usually an
extreme abundance.
D The reverse problem is why are intolerant
species intolerant ?
1 A physiological requirement for higher
oxygen levels is probably most basic.
2 Turbidity would hamper any organisms
employing sight in any way.
3 Absence of light would suppress the
growth of green algae, and hence also
restrict the growth of algae feeders.
E Inert silts by themselves have many
damaging effects such as abrasive or
smothering action.
Biological mechanisms for enduring inert
silt or sand pollution are not numerous,
and consequently such locations are
usually known as biological deserts.
Since some life exists even in deserts
however, a few forms may occasionally
be found. In general they are typical sand
or mud dwellers. Since the available
food in such a substrate is at best of a
very low order, inhabitants of these
Situations must either seek buried or
trapped food particles or capture food
from the passing waters.
1 If there is no BOD involved, and water
and oxygen circulate down into the
deposit, burrowing forms such as
certain mayflies, annelid worms,
ammocetes lamprey larvae, micro-
crustaceans, and others may burrow
down to depths of two feet or more.
Fish eggs normally deposited in gravel
and newly hatched larvae are also
dependent on circulating water. Such
a population can be killed overnight by
a layer of fine sediment or sludge which
seals the surface to water circulation.
2 Water or plankton feeders include
clams and mussels which can move
about freely m a soft, shifting bottom,
thus keeping on top of silt or sand as
-------
Effects of Pollution on Aquatic Life
it accumulates. If deposition is
actively taking place, however, there
will probably be so much turbidity in
the water that plankton (food) organisms
are unable to live. Under such cir"
cumstances certain clams and snails
have the ability to close the shell so
tightly by means of valves or an
operculum that all contact is lost with
the environment for extended periods,
during which COg tends to lower the
rate of body metabolism. Organisms
of this type have been reported to be
dug up with sand and gravel and
incorporated into concrete products
while still alive. Emergence of a
population of asiatic clams (Corbicula)
for example, just as a big block of
concrete is setting is said to be rough
on contractors'
3 An interesting situation occasionally
develops in estuaries where mud of
moderate organic content is slowly
deposited over oyster beds. The
oysters are unable to move, but as
they grow, their shells tend to bend
upward above the accumulating silt,
and may grow to several inches m
length while growing very little in
width. Crowding brings about a similar
reaction in the effort to avoid being
buried.
F Few generalizations can be offered
relative to toxic pollution except that
toxicity is relative, and all forms do not
respond equally to a given toxicant.
1 Few mechanisms of toleration can be
listed, beyond the natural resistance
that certain forms may have for a given
condition. For example, some marine
species may survive a salt concentration
that is toxic to freshwater species.
Over a period of several generations,
some species may develop a genetic
resistance to some toxicant such as
insecticides in the same way that DDT-
resistant strains of houseflies have
developed. Copper sulfate, and chlorine"
resistant strains of algae such as
Cosmanum for example, may develop
in treated water supply reservoirs.
2 Some bottom in-fauna organisms such
as annelid worms may retreat down
into burrows until a slug of undesirable
water passes.
3 Molluscs may close shells tightly for
the same purpose. The metabolic
rate is known to dimmish with the
increase of COg inside the closed
shell, thereby enabling them to
remain tightly sealed for extended
periods of time.
VII EFFECTS OF LIFE HISTORY STAGES
A In order to survive in a polluted area,
each life history stage of an indigenous
organism must be able to survive m turn.
B If some given life history stage cannot
tolerate conditions, and the species is
present:
1 Fortuitous changes may occur at the
proper time(s) to permit survival of
the more susceptible stages(s) or:
2 Recruitment from less polluted areas
may occur.
C Some examples of reproductive stages or
procedures which might affect pollution
sensitivity.
1 Egg or egg-like stages are often
enclosed in protective membranes,
jelly masses, or cases. May remain
dormant until favorable conditions
develop.
2 Eggs may be deposited in locations
where they are less exposed to polluted
water as:
buried in the gravel,
on the surface film,
on rocks over the water moistened
by spray,
on mud surface near water,
or in locations where maximum
water circulation is encountered
as at lip of waterfall.
-------
Effects of Pollution on Aquatic Life
3 Eggs may require minimal DO due to
low metabolic rate.
4 Eggs deposited on or in bottom may be
susceptible to smothering.
5 Newly hatched larvae often continue to
live on stored yolk material for a time.
On beginning to take natural food, they
may be killed by toxic content thereof,
such as organochlonnes.
6 Some forms such as certain sludge
worms commonly reproduce by
(vegetative) fragmentation, hence
avoiding egg and larva stages.
VIE NATURAL SELECTION AND
ACCLIMATIZATION TO POLLUTION
A Known biological mechanisms for selective
breeding of pollution resistant strains
operate in nature among fishes as among
other organisms.
1 Studies of population genetics indicate
that after some finite number of gen"
erations of population stress (e.g.,
exposure to a given pollutant),
permanent heritable resistance may be
expected to develop.
2 If the environmental stress (or
pollutant) is removed prior to the time
that permanent resistance is developed
in the population, reversion to the
non-resistant condition may occur within
a relatively few generations.
3 Habitats harboring populations under
stress in this manner are often marked
with the dead bodies of the unsuccessful
individuals.
B Individual organisms on the other hand can
over a period of time (less than one life
cycle) develop a limited ability to tolerate
different conditions, e.g., pollutants:
1 With reference to all categories of
pollutants both relatively facultative
and obligate species are encountered
(e. g., euryhalme vs stenohaline,
eurythermal vs stenothermal).
2 This temporary somatic acclimatization
is not heritable.
C A given single-species collection or
sample of living fishes may therefore
represent one or more types of pollution
resistance'
1 A sample of an original population
which has been acclimated to a given
stress in toto.
2 A sample of the surviving portion of
an originalpopulation, which has been
"selected" by the ability to endure the
stress. The dead fish in a partial fish
kill are that portion of the original
population unable to endure the stress.
3 A sample of a sub "population of the
original species in question which has
in toto over a period of several
generations developed a heritable
stress resistance.
D Any given multi-species field collection
will normally contain species illustrative
of one or more of the conditions outlined
above.
acknowledgment's-
Certain portions of this outline contain
material from a prior outline by Croswell
Henderson and revisions by R. M. Sinclair.
REFERENCES
1 Cordone, A.J. and Kelley, D. W. The
Influence of Inorganic Sediment on the
Aquatic Life in Streams. Calif. Fish
and Game. 47 189-228. No. 2. 1961.
2 Ellis, M.M. Detection and Measurement
of Stream Pollution. Bull. 22, U. S.
Bur. Fish: also, Bull. Burg. Fish 48.
356-437. 1937.
3 Foster, R.F. and Davis, J.J. The
Accumulation of Radioactive Sub-
stances in Aquatic Forms, No. A/Conf.
8/P/280. U.S.A. International Conf.
on Peaceful Uses of Atomic Energy,
pp. 1-7. 1955.
-------
Effects of Pollution on Aquatic Life
4 Ingram, W.M. and Towne, W.W.
Stream Life Below Industrial Outfalls.
Public Health Reports. 74 1059" 1070.
1959.
5 Kurland, Leonard. The Outbreak of a
Neurologic Disorder in Mimmata,
Japan, and its Relationship to the
Ingestion of Seafood Contaminated by
Mercuric Compounds. Proc. Nat.
Shell. San it. Workshop, pp. 225~228.
1961.
6 Mackenthun, K.M. and Keup, L. E.
Assessing Temperature Effects with
Biology. Proc. Am. Power Conf.
Vol. 31. pp. 335-343. 1969.
7 Tarzwell, C.M. and Gaufin, R.R. Some
Important Biological Effects of Pollution
Often Disregarded in Stream Surveys.
Purdue Univ. Engr. Bull. Proceedings
8th Ind. Waste Conf. May 4, 5, and 6,
1953.
8 Tarzwell, C.M. Hazards of Pesticides
to Fishes and the Aquatic Environment.
The Use and Effects of Pesticides.
Proc. of Symposium, Albany, N.Y.
Sept. 23, 1963. N.Y. State Joint
Legis. Comm. on Nat. Resources,
Albany, N.Y. pp. 30-40.
9 Vinson, S. B., Boyd, C. E. and Ferguson,
D.E, Resistance to DDT in the
Mosquito Fish Gambusia affinis Science.
139:217-218. January 18, 1963.
10 Walshe, Barbara M. On the Function of
Hemoglobin in Chironomus after Oxygen
Lack. Jour. Exp. Biol. Cambridge.
124:329-342. 1947.
SUPPLEMENTARY READING
1 Bullock, Glen L. A Schematic Outline for'
the Presumptive Identification of
Bacterial Diseases of Fish. Prog. Fish
Cult. 23(4).147-151. 1961.
2 Foster, R.F. and Davis, J.J. Aquatic
Life Water Quality Criteria. Second
Progress Report, Aquatic Life
Advisory Committee, Sewage and
Ind. Wastes, 28:678-690. 1956.
3 Fox, H. Munro. Daphma Hemoglobin.
Nature. London, p. 431.
September 27, 1947.
4 Ingram, W.M. and Wastler, III, T.A.
Estuarine and Marine Pollution.
Selected Studies, U.S. DHEW, PHS,
Robt. A. Taft Sanitary Engineering
Center, Cincinnati, Ohio. Technical
Publication No. W6 l-04.
5 Jackson, H.W. and Brungs, Wm. A.
Biomonitoring of Industrial Effluents.
Purdue Industrial Waste Conference,
Layfayette, Indiana. May 3*5, 1966
6 Rodhe, W, Limnology, Social Welfare,
and Lake Kinneret. Int. Jour.
Limnology, Vol. 17. November 1969.
7 Tennessee Valley Authority, Fish Kill
in Boone Reservoir. TVA Water
Qual. Branch. Chattanooga, Tenn.
1968.
8 Robert A. Taft Sanitary Engineering
Center. Pesticides in Soil and Water.
An annotated Bibliography. PHS
Publication No. 999-WP-17.
September 1964.
9 Stewart, R. Keith, Ingram, William M.
and Mackenthun, Kenneth. Selected
Biological References on Fresh and
Marine Waters. FWPCA Publication
No. WP-23, pp. 126. 1966.
10 Warren, Charles E. Biology and Water
Pollution Control. W.B. Saunders Co.
434 pp. 1971.
This outline was prepared by H.W. Jackson,
Chief Biologist, National Training Center,
WPO, EPA, Cincinnati, OH 45268.
-------
THE EFFECTS OF ORGANISMS ON POLLUTION AND THE ENVIRONMENT
I INTRODUCTION
A Pollution is often studied as a factor
affecting the biota, but it is equally
important to recognize the environmental
changes produced by the biota.
B According to Westlake, under many
conditions . . . the environment is almost
as much a product of the community as the
community is of the environment. "
II SOME ENVIRONMENTAL EXAMPLES
A Diatom Blooms
"... Astenontella (an oil storage alga)
produces an autotoxin (autoantibiosis)
which will inhibit itself, may stimulate or
inhibit other species that store oil, but
always stimulate algae that store starch.
For example, Asterionella may produce
a bloom and inhibit itself, but stimulate
a population of Synedra. These oil
storage algae produce a substance that
stimulates a starch-storing alga,
Coelastrum, which may stimulate another
starch-storing organism, Cosmanum, and
they, in turn, stimulate an oil storing
species of Dinobryon. " Patrick.
B The altered structure of the plankton
community due to the introduction of the
alewife
C Fecal deposit feeders in the estuanne
environment
D Particle feeders are successful in the
pipe clogging community and are
generally (Sebestyen)-
1 Sessile
2 Suspension feeders
3 Have motile larvae
4 Resistant stages
E Biogeochemical Cycles
In terms of biomass and energy flow, the
mussel Modiolus (Figure 1) is a relatively
minor component of the marsh community.
However, they have been demonstrated to
have a major effect on the recycling and
retention of valuable phosphorus, thereby
maintaining fertility and production of
autotrophs (Odum).
F Sudd (dense aggregations of floating weeds)
Flowering aquatic plants are a serious
problem in shallow, stagnant, or slow-flowing
water in many tropical countries. They are
expensive liabilities in newly impounded
reservoirs in developing nations.
G Biological Pollution
Contamination of living native biotas by
introduction of exotic life forms has been
called biological pollution by Lachner et al.
Some of these introductions are compared
to contamination as severe as a dangerous
chemical release. They also threaten to
replace known wildlife resources with
species of little or unknown value.
1 Tropical areas have especially been
vulnerable. Florida is referred to as
"a biological cesspool of introduced
life".
2 Invertebrates
a Asian Clams have a pelagic veliger
larvae, thus, a variety of hydro
installations are vulnerable to sub-
sequent pipe clogging by the adult
clams.
b Melanian snails are intermediate
hosts for various trematodes
parasitic on man.
3 Vertebrates
a At least 25 exotic species of fish
hcve been established in North
America.
b Birds, including starlings and cattle
egrets.
WP. NAP. 22 d. 9.72
-------
The Effects of Organisms on Pollution and the Environment
c Mammals, including nutria.
4 Aquatic plants
Over twenty common exotic species are
growing wild in the United States. The
problem of waterway clogging has been
especially severe in parts of the
Southeast.
IE In polluted environments, there is a more
noticeable change in balance, time response,
and effects.
A Organic Pollution
The conditions here are classic and well
described. There is a succession of
biological communities (Figure 2) each
of which modify the environment and water
properties, thus, the effects are pre-
dominately biological.
1 Tubificidae (aquatic earthworms) m the
first zone may reach as many as
1, 7000, 000/m and move up to 50 tons
of mud per acre per day.
2 In the second zone chironomids (midge
larvae) are found in thousands per
square meter and may reduce the DO
level by one and one-half ppm per
stream mile.
3 The Isopod (sowbug) zone (the genus
Asellus or Lirceus depending on locality)
third in succession also may reach a
density of thousands per square meter,
a further oxygen demand due to
respiration. (Figure 2)
4 The filamentous green alga Cladophora
in both streams and lakes responds to
organic enrichment by producing dense
growths. (Figure 3)
5 Higher aquatic plants such as
Potamogeton pectinatus may also be
involved in these ecological changes
in streams, particularly respiration
vs. photosynthesis. There is correlation
between weed bed growths, velocity and
silt deposition. In some streams these
massive growths on sloughing off foul
water intakes and reduce DO levels as
they decompose.
6 Sphaerotilus and/or slime growths
below organic wastes, by metabolic
demands while living and decomposition
after death, impose a high BOD load
on the stream and can severely deplete
the dissolved oxygen. (Figure 3)
7 Blackflies (Diptera, family Simuliidae)
often reach large populations below
organic waste sources, filter feed on
this material and in so doing, further
degrade stream conditions with their
fecal deposits.
B Inorganic and Toxic Pollution
When progressive changes occur they
are rarely produced by the biological
community.
C Biological Magnification
Biological magnification is an additional
chronic effect of toxic and other pollutants
(such as heavy metals, pesticides,
carcinogens, teratogens, radionuclides,
bacteria, and viruses) which must be
recognized and examined before clearance
can be given for the disposal of a waste
product into natural waters. To para-
phrase Odum we could give nature an
apparently innocuous amount of pollutant
and have her give it back to us m a lethal
or detrimental package.
1 Many animals, and especially bivalves,
have the ability to remove from the
environment and store in their tissues
substances present at nontoxic levels
in the surrounding water
a This process may continue in the
clam or fish, for example, until
the body burden of the toxicant
reaches such levels that the animal's
death would result if the pollutant
were released into the bloodstream
by physiological activity.
-------
The Effects of Organisms on Pollution and the Environment
autotrophs ^
^^Sportina-
benthic algae
PHOSPHORUS IN WATER
particulate Wmgm/m'
phosphate 19
dissolved organic 6
total 39 mgrn/m1
5.5 mgm/mVday
intak#
sedimented
5.2 mgm /m:/day
g " ¦ iTTl^Cu
i
i
0.3 mgm/mVdoy
recycle
MODIOLUS
POPULATION
—^ ENERGY FLOW -
flux by Modiolus
amount in environment
phosphorus 37 mgm/m1
BIOMASS lMg/m*
respiration 0.1 kcal/m1 day
^ production 0.05 "
-0.15 —
Ppa^O.37
•nerjy : : 0.008
(Reproduced from Figure 4-4, from Ecology by Eugene P. Odum,
copyright (c) 1963 by Holt, Rinehart and Winston, Inc. used by
permission of Holt, Rinehart and Winston, Inc.)
Figure 1. The role of a shellfish (mussel) population
in the cycling and retention of phosphorus
in an estuarine ecosystem.
MILES
Figure 2. Linear alterations in populations of Tubificids (A)
Chironomids (B), and Isopods (C) (from Bartsch).
Zone C is often referred to as the Cladophora-
Asellus Zone.
-------
The Effects of Organisms on Pollution and the Environment
10 0 10 20 30 40 50 60
Miles
Figure 3. Linear alterations in populations of slime growths and Cladophora
(modified from Bartsch)
b This may occur, as in the case of
chlorinated hydrocarbon pesticides
(such as DDT and endrin) stored in
fat depots, when the animals food
supply is restricted and the body
fat is mobilized.
c The appearance of the toxicant in
the bloodstream causes the death
of the animal.
2 The biological magnification and
storage of toxic residues of polluting
substances and microorganisms may
have another serious after effect.
a Herbivorous and carnivorous fish at
lower trophic stages may gradually
build up DDT residues of 15 to 20
mg/1 without apparent ill effect.
b Carnivorous fish, mammals, and
birds preying on these contaminated
fish may be killed immediately or
suffer irreparable damage because
of the pesticide residue or infectious
agent.
c Commercial shellfishenes are
damaged by toxms produced by some
dino-flagellate blooms m nearshore
waters.
IV In summary, the biological causes of DO
changes, associated changes in pH and CO^,
ammonia, nitrates, and sulphides, require
study if the effects of organic pollution are
to be calculated and predicted. Further
biological effects on pollution include the
relation to fouling organisms, stabilization
of sediments in estuaries, recycling of
nutrients, and problems of biological
magnification.
REFERENCES
1 Curtis, E.J. Review Paper. Sewage
Fungus Its Nature and Effects.
Water Res. 3-289-311. 1969.
2 Herbst, Richard P. Ecological Factors
and the Distribution of Cladophora
glome rata in the Great Lakes. Amer.
Mid. Nat. 82( 1):90-98. 1969.
3 Holm, L. G., Weldon, L.W. and Blackburn,
R.D. Aquatic Weeds. Science.
66-699-709. 1969.
4 Odum, E.P. Ecology. Holt, Rinehart,
and Winston. 192 pp, 1961.
-------
The Effects of Organisms on Pollution and the Environment
5 Lachner, Ernest A., Robins, C. Richard,
and Courtenay, Walter R. Jr.
Exotic Fishes and Other Aquatic
Organisms Introduced into North
America Smithsonian Contrib. to
Zool. 59:1-29. 1970.
6 Patrick, Ruth. Water Research Programs
Aquatic Communities. Office Water
Resources, U.S. Department of the
Interior, Washington, D. C. 22 pp. 1968.
7 Sculthorpe, C.D. The Biology of Aquatic
Vascular Plants. St. Martin's Press,
New York. 610 pp. 1967.
/
8 Sebestyen, Olga. On Urnatella gracilis
Leidy (Kamptozoa Cori) and its
occurrence in an industrial waterworks
fed by Danube water in Hungary. Acta
Zool. Acad. Sci. Hungaricae.
8.435-448.
9 Westlake, D. F. The Effects of Orga-
nisms on Pollution. Proc. Linnean
Soc. London. 170 session pt. 2.
p. 171-172. 1959.
10 Westlake, D.F. The Effects of
Biological Communities on Conditions
in Polluted Streams. Symp. No. 8
Inst, of Biol. London (41 Queen's
Gate, London, S.W.T.)p. 25-31. 1959.
11 Whitton, B, A. Review Paper, Biology
of Cladophora in Freshwa+ers, Water
Research 4:457-476. 1970.
This outline was prepared by Ralph M.
Sinclair, Aquatic Biologist, National
Training Center, Water Programs Operations,
EPA, Cincinnati, OH 45268.
-------
THE EFFECTS OF POLLUTION ON LAKES
I INTRODUCTION
The pollution of lakes inevitably results in a
number of undesirable changes in water
quality which are directly or indirectly
related to changes in the aquatic community.
A Industrial Wastes may contain the following-
1 Sewage
2 Dissolved organics--synthetics, food
processing wastes, etc.
3 Dissolved minerals--salts, metals
(toxic and nontoxic), pigments, acids, etc.
4 Suspended solids--fibers, minerals,
degradable and non-degradable organics
5 Petroleum products--oils, greases
6 Waste heat
B The Materials in Domestic Wastes which
affect Water Quality are
1 Pathogenic fecal microorganisms
2 Dissolved nutrients, minerals, vitamins,
and other dissolved organic substances
3 Suspended solids (sludge)--degradable
and non-degradable organic materials
C Pollution and Eutrophication
The discharge of domestic wastes often
renders the receiving water unsafe for
contact water sports and water supplies.
For example, some beaches on the eastern
seaboard and in metropolitan regions of
the Great Lakes are unfit for swimming
because of high coliform counts. Other
effects of domestic pollution include
changes m the abundance and composition
of populations of aquatic organisms.
1 As the nutrient level increases, so does
the rate of primary production.
2 Shore-line algae and rooted aquatics
become more abundant. For example,
problems have been experienced with
Cladophora and Dichotomosiphon along
the shores of Lakes Ontario, Erie,
and Michigan. These growths interfere
with swimming, boating, and fishing,
and cause odors when the organisms
die and decay.
3 The standing crop of phytoplankton
increases, resulting in higher counts
and greater chlorophyll content.
Increases m phytoplankton abundance
may result in taste and odor problems
in water supplies, filter clogging,
high turbidity, changes m water color,
and oxygen depletion in the hypolimnion.
4 Populations of fish and larger swimming
invertebrates increase, based on the
increase in basic food production.
5 Changes in dominant species
a Diatom communities give way to
blue-greens. Toxic blue-greens may
pose a problem.
b Zooplankton changes include
replacement of Bosmina coregoni
by B. longirostris.
c Trout and whitefish are replaced by
perch, bass, and rough fish.
d Hypolimnion becomes anaerobic in
summer, bottom sludge buildup
results in loss of fish food organisms,
accompanied by increase in density
of sludgeworms (oligochaeta).
II HISTORICAL REVIEW
The cultural eutrophication of a number of
lakes in Europe and America has been well
documented.
A Zurichsee, Switzerland
WP. LK. lc. 4.70
-------
The Effects of Pollution on Lakes
1 1896 - sudden increase in Tabellaria
fenestrata
2 1898 - sudden appearance of Oscillatoria
rubescens which displaced Fragilaria
capucina
3 1905 - Melosira lslandica var. helvetica
appeared
4 1907 - Stephanodiscus hantzschu
appeared
5 1911 - Bosmina Ion giro str is replaced
B. coregoni
6 1920
1924 - O. rubescens occurred in great
quantities
7 1920 - milky-water phenomenon,
precipitation of CaCO^ crystals (40pi)
due to pH increase resulting from
photosynthesis
8 Trout and whitefish replaced by perch,
bass, and rough fish
B Hallwilersee, Switzerland
1 1897 - Oscillatana rubescens not
observed up to this time
2 1898 - O. rubescens bloomed,
decomposed, formed H^S, killing off
trout and whitefish
C Lake Windermere, England (core study)
1 Little change in diatoms from glacial
period until recent times
2 Then Astenonella appeared, followed
by Synedra
3 About 200 years ago, Asterionella
again became abundant
4 Asterionella abundance ascribed to
domestic wastes
D Finnish Lakes
Aphanizomenon, Coeloaphaerium,
Anabaena, Microcystis, are the most
common indication of eutrophy.
TABLE 1 CHANGES IN PHYSIO-CHEMICAL PARAMETERS
Zurichsee, Switzerland
Parameter Date
Chlorides
Dissolved organics
1888
1916
1888
1914
Value
1. 3 mg/1
4. 9 mg/1
9. 0 mg/1
20. 0 mg/1
Secchi Disk
Dissolved oxygen, at
100 M, mid-summer
before 1910
1905 - 1910
1914 - 1928
1910 - 1930
1930 - 1942
Max.
16.8M
10. 0M
10. 0M
Minimum
Min.
3.1M
2.1M
1.4M
100% saturation
. 9% saturation
-------
The Effects of Pollution on Lakes
E Linsley Pond, Connecticut
1 Species making modern appearance
include Asterionella formosa,
Cyclotella glomerata, Melosira
italica. Fragilaria crotonensis,
Synedra ulna
2 Asterionella formosa and Melosira
italica were considered by Patrick to
indicate high dissolved orgamcs
3 Bosmina coregoni replaced by B.
longirostris
F Lake Monona, Wisconsin
1 Began receiving treated sewage in 1920,
developed blue-green algal blooms.
G Lake Washington, Washington
1 1940 - Bosmina longirostris appeared
2 1955 - Oscillatoria rubescens seen for
the first time, and constituted 96% of
phytoplankton, July 1
H Lake Erie
1 Phytoplankton counts at Cleveland have
increased steadily from less than
500 cells/ml in the 1920's to over
1500 cells/ml in the 1960's
2 Abundance of burrowing mayflies
(Hexagenia spp.)in Western Lake Erie
decreased from 139/m2 in 1930, to
less than 1/m2 in 1961.
I Lake Michigan
1 Milky water observed in south end, and
in limnetic region in mid-1950's and
again m 1967.
2 During the period 1965-1967 the Chicago
water treatment plant has found it
necessary to increase the carbon dosage
from 23 lbs/mil gal to 43 lbs/mil gal,
and the chlorine dosage from 20 lbs/mil
gal to 25 lbs/mil gal.
3 Phytoplankton counts in the south end
now exceed 10, 000/ml during the
spring bloom.
III FACTORS AFFECTING THE RESPONSE
OF LAKES TO POLLUTION INCLUDE.
A Depth-surface area ratio- A large
hypolimnion will act as a reservoir to
keep nutrients from recirculating in the
trophogenic zone during the summer
stratification period. Rawson found an
inverse relationship between the standing
crop of plankton, benthos, and fish, and
the mean depth.
B Climate- Low annual water temperatures
may restrict the response of the
phytoplankton to enrichment.
C Natural color or turbidity: Dystrophic
(brown-water) lakes may not develop
phytoplankton blooms because of the low
transparency of the water.
IV TROPHIC LEVEL
Except in cases where massive algal blooms
occur, the trophic status of lakes is often
difficult to determine. Core studies are
used to determine trends in diatom populations
which might indicate changes in nutrient
levels over an extended period of time.
V CONTROL OF POLLUTION
The success of efforts to arrest the
eutrophication process, and where desirable,
reduce the trophic level of a lake, will
depend on a thorough knowledge of the
nutrient budget.
A Significant quantities of nutrients may
enter a lake from one or more of the
following sources-
1 Rainfall
2 Ground water
-------
The Effects of Pollution on Lakes
TABLE 2 PARAMETERS COMMONLY USED TO DESCRIBE CONDITIONS
1 Transparency
2 Phosphorus
3 NOg - Nitrogen
4 Minimum annual
hypolimnetic oxygen concentration
5 Chlorophyll
6 Ash-free weight of seston
7 Phytoplankton count
8 Phytoplankton quotients
a number of species of Chlorococcales
number of species of Desmids
Oligotrophic Condition
> 10 meters
< lug/1
< 200 ug/1
near 100% saturation
, 3
< 1 mg/m
< 0.1 mg/1
< 500/ml
<1
b Myxophycease+Chlorococcales+Centrales+Euglenaceae <1
Desmidaceae
c Centrales 0 - 0. 2
Pennales
9 Phytoplankton species present (see outline on
plankton in oligotrophic lakes).
3 Watershed runoff
4 Shoreline domestic and industrial outfalls
5 Pleasure craft and commercial vessels
6 Waterfowl
7 Leaves, pollen, and other organic
debris from riparian vegetation
B The supply of nutrients from "natural"
sources in some cases may be greater
than that from cultural sources, and be
sufficient to independently cause a rapid
rate of eutrophication regardless of the
level of efficiency of treatment of domestic
and industrial wastes.
C Many methods have been employed to
treat the symptoms, reduce the
eutrophication rate, or completely
arrest and even reverse the eutrophication
process.
1 Use of copper sulfate, sodium arsenite,
and organic algicides- It is not
economically feasible to use algicides
in large lakes.
2 Addition of carbon black to reduce
transparency. This is likewise
frequently impractical.
3 Harvesting algae by foam fractionation
or chemical precipitation.
-------
The Effects of Pollution on Lakes
4 Reducing nutrient supply by (a) removal
of N and P from effluents, (b) diversion
of effluents, and (c) dilution with
nutrient-poor water.
D Examples of lakes where control has been
attempted by reducing the nutrient supply,
are1
1 Lake Washington, Seattle
The natural water supply for this lake
is nutrient poor
(Ca = 8 mg/1, P < 5 Mg/1, TDS = 76mg/l).
Since the turnover time of the water in
this lake is only three years, it was
expected that diversion of sewage
would result in a rapid improvement of
water quality. Diversion began in 1963,
and improvements were noticeable by
1965 - including an increase in
transparency, and a reduction in seston,
chlorophyll, and epilimnetic phosphorus.
TABLE 3
PHOSPHORUS REDUCTION IN LAKE WASHINGTON
Maximum phosphorus in
upper 10 meters
Year (Mg/1)
1963 70
1964 66
1965 63
2 Green Lake, Washington
The lake has a long history of heavy
blooms of blue-green algae. Beginning
in 1959, low-nutrient city water was
added to the lake, reducing the con-
centration of phosphorus by 70% in the
inflowing water. By 1966, the lake had
been flushed three times. Evidence of
improvement in water quality was noted
in 1965, when Aphanizomenon was
replaced by Gleotrichia.
3 Lake Tahoe
This lake is still decidedly oligotrophy.
To maintain its high level of purity,
tertiary treatment facilities were
installed in the major sewage treat-
ment plant, and construction is now
underway to transport all domestic
wastes out of the lake basin.
REFERENCES
1 Ayers, J. C and Chandler, D. C , Eds
Studies on the environment and
eutrophication of Lake Michigan.
Special Report No. 30 Great Lakes
Research Division, Institute of
Science and Technology, University
of Michigan, Ann Arbor. 1967
2 Brezonik, P. L , Morgan, W.H.,
Shannon, E.E , and Putnam, H.D.
Eutrophication factors in North
Central Florida Lakes University
of Florida Water Res. Center
Pub #5, 101 pp. 1969.
3 Carr, J. F , Hiltunen, J. K Changes
in the bottom fauna of Western Lake
Erie from 1930 to 1961 Limnol
Oceanogr 10(4) 551-569. 1965.
4 Frey, David G. Remains of animals
in Quaternary lake and bog sediments
and their interpretation.
Schweizerbartsche Verlagsbuchhandlung.
Stuttgart. 1964
5 Edmondson, W.T , and Anderson, G.C.
Artificial eutrophication of Lake
Washington. Limnol. Oceanogr
1(1) 47-53 1956.
6 Fruh, E.G. The overall picture of
eutrophication. Paper presented
at the Texas Water and Sewage
Works Association's Eutrophication
Seminar, College Station, Texas.
March 9, 1966.
7 Fruh, E.G , Stewart, K.M , Lee, G.F
and Rohlich, G.A. Measurements
of eutrophication and trends.
J.W.P.C.F. 38(8)-1237-1258 1966
-------
The Effects of Pollution on Lakes
8 Hasler, A.D. Eutrophication of lakes
by domestic drainage. Ecology
28(4) 383-395. 1947.
9 Hasler, A.D. Cultural Eutrophication
is Reversible. Bioscience 19(5):
425-443. 1969.
10 Herbst, Richard P, Ecological Factors
and the Distribution of Cladophora
glome rata in the Great Lakes.
Amer, Midi. Nat. 82(l):90-98. 1969.
11 National Academy of Sciences.
Eutrophication- Causes, Consequences,
Correction. 661 pp. 1969.
(Nat. Acad. Sci. ,2101 Constitution
Avenue, Washington, DC 20418, 13.50).
12 Neel, Joe Kendall. Reservoir
Eutrophication and Dystrophication
following Impoundment. Reservoir
Fisheries Res. Symp. 322-332.
13 Oglesby, R.T. and Edmondson, W.T.
Control of Eutrophication.
J.W.P.C.F. 38(9):1452-1460. 1966.
14 Stewart, K. M. and Rohlich, G.A.
Eutrophication - A Review.
Publication No. 34, State Water
Resources Control Board, The
Resources Agency, State of California.
1967.
This outline was prepared by C. I. Weber,
Chief, Biological Methods Section,
Analytical Quality Control Laboratory,
NERC, EPA, Cincinnati, OH 45268.
-------
WATER TEMPERATURE AND WATER QUALITY
I INTRODUCTION
A Temperature is the basic variable m water
"climates".
1 Temperature, or the amount of thermal
energy present, is originally of solar
or cosmic origin. Biological processes
acting over geologic time temporarily
capture and store much energy in
organic substance. The "fossil fuels"
(oil, gas, and coal) which we burn
today release solar energy captured
in the geologic past, which but for man
might not have been released until
sometime in the geologic future. Most
of the solar energy being captured
today, is released probably today,
with the possible exception of some
unrecognized fraction which is pro-
ceeding into long term storage.
The release of atomic energy is of
inorganic or cosmic origin, and the
magnitude and significance of the
additional thermal discharge to the
environment has yet to be accessed.
One last observation is important
before turning to the details of water
temperature and water quality. While
we are most critically occupied with
the immediate or local impact of a
concentration of thermal energy released
at a given point in the environment
(the excess or "waste" which manisunable
to capture and entrain in his electric
transmission lines), it should be
remembered that of every ton of coal
or pound of uranium burned as fuel,
nearly 100% of the energy contained
eventually finds its way into the general
global environment. A small
remaining fraction is rebound as
chemical energy in some "product. "
2 Direct solar radiation is the overriding
contributor of thermal energy to all
lands and waters.
a Total energy from insolation onto
"spacecraft earth" is counter-
balanced by the radiation of
terrestrial energy into space.
If the two do not exactly balance
on an annual basis, the overall
temperature (climate) of the earth
will rise or fall.
b The annual climate or heat budget
of a given body of water is deter-
mined by its geographic location
(latitude elevation, etc.) interacting
with local meteorological conditions,
and other factors.
c There is, therefore, a natural or
normal temperature regimen for
any given body of water to which
it will tend to return if disturbed
by man.
d There is, also, a normal or
characteristic community of
aquatic organisms that will tend
to persist. When the heat budget
or climate of a body of water is
changed, the fauna and flora change.
3 There is great diversity of opinion,
even among knowledgeable people, as
to the effects of thermal changes m
waters. Some of the reasons for this
follow •
a Only a continuously maintained
temperature of 100° C could keep
a surface water mass sterile.
The question is therefore not life or
no-life; but rather- what kind of
life is the objective9
BI. ECO. he. lb. 4. 70
-------
Water Temperature and Water Quality
b Aquatic life has received more
attention than other water uses
because the aquatic organisms
cannot escape the water conditions.
c There are certain circumstances in
which a modest rise in temperature
might be considered to be beneficial
as for example: keeping an area of
a river or a harbor free of ice for
navigation or winter fishing.
There has, also, been investigation
of the use of warmed waters for
certain aspects of aquaculture.
d It is, therefore, important in
discussing the virtues and vices
of thermal changes to clearly
define or specify the objective or
type of aquatic community in mind.
4 It is clear that the need for more and
more power will continue into the
foreseeable future (Table 1).
B Human activities which may modify
receiving water temperatures include
the following-
1 Logging and other land stripping
activities which increase the rate of
surface run-off and hence raise or
lower temperatures of influent waters,
depending or\ the season.
2 Removal of stream bank shade
II THERMAL ELECTRIC POWER
PRODUCTION AS A STREAM WARMING
ELEMENT
A Production of electric power by stream
plants involves the wastage of considerable
quantities of energy in cooling waters.
Approximately 5000 BTU of heat are
wasted for each kilowatt of electricity
generated. This represents an efficiency
of roughly 40%.
B It is estimated that approximately 80%
of all energy required in the future will
come from steam generating plants.
C Weirs and jetties help greatly in the
dispersal of warmed waters, but must
be carefully designed to each situation.
D As water temperature rises, its value as
a coolant diminishes.
E Heat dissipation from a body of water
which has been heated above its equilibrium
temperature with the meteorological con-
ditions follows Newton's Law of Cooling
which states that the rate of cooling is
proportional to the difference between
the temperature of the body of water,
and the equilibrium temperature for the
given meteorological conditions. For
example, an analysis of the Ohio River
as at Cincinnati has shown that it would
require over 200 miles to dissipate
99 + % of heat added (Figure 1).
3 Erosion which fills in stream bed and
causes water to be spread in broad
shallow layer, exposed to sun and air.
4 Release of cold waters from hypolimnion
of deep reservoirs.
5 Withholding or augmenting flow by dam
manipulation.
6 Release of relatively large volumes of
high temperature wastewaters from
power production and/or industrial
processes.
Ill EFFECTS OF HEAT ON ORGANIC
WASTE DISPOSAL
A Higher temperatures accelerate the rate
of bacterial growth, the optimum tem-
perature being in the range of 30° C
(86°F),
1 As water temperatures approach this
value, the rate of BOD thus approaches
a maximum.
-------
Water Temperature and Water Quality
TABLE 1
MAXIMUM TEMPERATURES PROBABLY COMPATIBLE WITH THE WELL-BEING
OF VARIOUS SPECIES OF FISH AND THEIR ASSOCIATED BIOTA IN o C
Temperature Taxa
34 C Growth of catfish, gar, white or yellow
bass, spotted bass, buffalo, carpsucker,
threadfin shad, gizzard shad
32 C Growth of largemouth bass, drum, bluegill,
crappie
29 C Growth of pike, perch, walleye, smallmouth
bass, sauger, California killifish, topsmelt
27 C Spawning and egg development of catfish,
buffalo, threadfin shad, gizzard shad,
California grunion, opaleye, northern
swellfish
24 C Spawning and egg development of large "
mouthed bass, white and yellow bass, spotted
bass, sea lamprey, alewife, striped bass
19 C Growth or migration routes of salmonoids
and for egg development of perch, small-
mouth bass, winter flounder, herring
12 C Spawning and egg development of salmon
and trout (other than lake trout)
9 C Spawning and egg development of lake trout,
walleye, northern pike, sauger, and Atlantic
salmon
2 If the waste assimilative capacity of
a stream is being utilized based on a
given stream temperature, and the
temperature subsequently raised, the
DO may drop so low as to produce fish
kills and other nuisance conditions.
B Higher water temperatures may, also,
lead to a higher concentration of bacteria
pathogenic to man.
IV EFFECTS OF HEAT ON FISH AND OTHER
AQUATIC LIFE
A Vulnerability of fish and other aquatic life
to high temperatures represents a major
restriction on the discharge of cooling
water.
1 Involves duration of exposure as well
as absolute thermal level.
2 Sensitivity to toxic substances is
increased.
3 Lower temperatures are required in
winter than in summer.
B Factors contributing to the sensitivity
of fish to heat
1 Oxygen solubility diminishes as
temperature rises (Figure 2).
2 Oxygen requirements of aquatic life
increase as temperature rises
(Figure 3).
-------
Water Temperature and Water Quality
x o 2
UJ
MARKUND
DAM ~
MILES FROM POINT OF DISCHARGE
TEMPERATURE DROP THROUGH MARKLAND POOL
OHIO RrVER
Figure 1
9 10 II 12
DISSOLVED OXYGEN p.p.m.
OXYGEN SOLUBILITY AT SELECTED SALINITIES
RICHARDS AND CORWIM, LIMNOLOGY AND OCEANOGRAPHY, 1956
BI.EC0.ec.2-B8
Figure 2
-------
Water Temperature and Water Quality
Act ive
t-- o
-C (\j
Standard
Scope
10 20 30
Temperature °C
40
THE RELATION OF TEMPERATURE TO ACTIVE AND STANDARD
METABOLISM IN YOUNG GOLDFISH OF AN AVERAGE WEIGHT OF
2 GM. From Fry and Hart (1948)
Figure 3
-------
Water Temperature and Water Quality
3 The sensitivity of aquatic organisms
to temperature levels and changes
varies with age, size, and size.
a A constant elevated temperature
reduces the potential of fish to
reproduce (Figure 4).
EFFECTS OF CONSTANT TEMPERATURE
ON REPRODUCTION OF A MINNOW
(Pimcphales promelas)
Figure 4
b Different species have different
preferred temperature ranges.
c Seasonal cooler temperatures are
often essential to egg production
and hatching, while warmer summer
temperatures will promote faster
growth after hatching, up to some
limit of tolerance characteristic of
the species.
4 Lethal high temperatures as deter"
mined in laboratory tests vary widely
for different species, e.g. goldfish.
107.6°F, pink salmon- 750F.
a Lethal temperatures differ at
different times of the year, as well
as for the different life history
stages (Figure 5).
b This is analogous to a temperature
of 50OF for man- in winter it feels
"warm, " in summer it is "cool. "
5 Sudden changes in water temperature
can be fatal to certain organisms, both
fish and fish food organisms.
6 Acclimatization to higher temperatures
is faster than to lower. Fish accli"
mated to warm water are rapidly
killed when they swim into cold water.
This implies that the sudden shutdown
of a thermal discharge may be more
detrimental than a continuous normal
discharge.
7 Reduction m DO, increase in CO^, or
the presence of toxic materials
reduces maximum tolerable tempera"
tures.
8 Species can be eliminated at less than
lethal temperatures by predators,
parasites, or diseases which are less
temperature "sensitive.
9 Some fish do not seem to be able to
avoid killing hot waters, while others
do.
10 Preferred temperature ranges in
laboratory tests generally are some"
what higher than in field observations
(Figure 6). This may be influenced
by the demands of the natural environ-
ment for greater activity and hence a
need for more oxygen (Figure 2, Table 1).
11 Temperature can act as a directive
force in fish migration.
12 The exact physiological mechanisms
of heat kill are not fully understood.
Fats rather than proteins seem to be
the most critical substance.
a Some fish will die at temperatures
of 65° F, lower than that at which
proteins usually coagulate.
b Tropical (or heat-adapted) plants
and animals often have fats with a
higher melting point than arctic
(or cold-adapted) organisms.
c Animals (such as goldfish) fed high
melting point fats (e.g. beef) develop
higher melting point body fats than
those fed on low-melting fats
(such as fish oil). They are in turn
able to tolerate higher temperatures.
-------
Water Temperature and Water Quality
100
80
60
40
20
0
100
80
60
40
20
0
°F)
BROOK TROUT
LETHALx
- \
nsafe
MATURATION
SPAWN -
INS
INCUBATION
FRY
GROWTH
°F] 10)20 IP> I
1 LETHAL x
FATHEAD MINNOW ^safe
MATURATION SPAWNING INCUBATION FST
GROWTH
-IPIIl 1 SCPTCHBEB I-
-SEf I XPB 1-
KNOWN
UNKNOWN
THERMAL TOLERANCE OF CRITICAL LIFE HISTORY STAGES
Figure 5
d Lethal temperatures seem to destroy
fat-calcium relationships.
C Effects of Temperature on Fish Food
Organisms
1 Species composition and abundance are
affected in ways similar to the fishes
as outlined above.
2 Warm waters encourage blue"green
algae. Some can tolerate as high as
185°F for limited periods (Figure 7).
3 A temperature increase of 8°C
stimulated photosynthesis in phyto-
plankton in
-------
Water Temperature and Water Quality
34
32
30
U
o
28
' 26
IE 24
z>
h
< 22
tr
UJ
| 20
UJ
h IB
16
14
12
I 0
LABORATORY STUDIES
liWIS MACROCMIRUS
Cyprinus ca®pio
UPOMfS CI8BOSUS
_ CARASSIU5 AURATVS
— Eso* verficulatus
Esox masouinongy .
GlRELLA NICR I CANS ¦
PERCA FLAVESCENS
— Oncorhynchus KETA ¦
ONCORHYNCHUS TSHAWYTSCHA
Oncorhynchus gossuscha
HCROPTERUS SALMOI OES ^
MlCROPTERUS DOLOMIEU
Perca FLAVESCENS
IOTA LOTA (Y0UN6 OF YEAR).
Salvelinus fontinalis-
C'JCORKYNCHUS NtRKA .
COREGOIAJS CLUPEAFORMIS
SALVElINUS HYBRID (F.X 1.
FIELD
OBSERVATIONS
MCROPTERUS SALMOIDES
MlCROPTERUS DOLOMIEU
perca flavescens
SaLVELINUS FONTINAUS
—Salvelinus hybrid
¦ COREGONUS CLUPEAFORMIS
-lota iota (yearling or/
¦Oncorhynchus nerka older)
Left. A comparison of various final preferenda as found by laboratory studies.
Right. A comparison of field observations with laboratory results for a number of species.
Figure 6
-------
Water Temperature and Water Quality
OIATOMS
r
GrtCNS
V FlUE
AcrifNS
\
68* 77" 66' 95" 104"
TEMPERATURE (F)
E The distribution of many benthic
invertebrate organisms is temperature
dependent(See Table 2).
ENVIRONMENTAL TEMPERATURE RANGES
OF SOME MARINE INVERTEBRATES
Table 2
Taxa
Temperature range inOC
American Oyster
4-34
European Oyster
0-20
Opossum Shrimp
0-31
EFFECT OF TEMPERATURE ON TYPES
OF PHYTOPLANKTON
Figure 7
F Observations in Miami, Florida indicate
that the following groups of larger plants
may show temporary or permanent
changes following thermal discharge:
1 The American Oyster Crassostrea
Virginicamav spawn, depending on its
condition, at temperatures from 15
to 340 C, spawning being triggered by
a rise in temperature.
2 The shore crab Carcides Maenas thrives,
but does not breed at temperatures
between 14 and 28QC. Breeding can
only take place outside the heated area.
C Fish in the estuarine environment are more
susceptible to temperature changes than
those in fresh water. However, wider
ranges of tolerance between species exist.
D Most shellfish (in the broad sense:
mollusca and crustaceans) are relatively
or highly stenothermal (unadaptable to
rapid temperature changes). Some are
stenothermal for one stage (e.g. spawning),
and eurythermal for others (e.g. growing).
1 The sea grass Thalassia, an important
habitat for invertebrates and stabilizes
of the substrate.
2 Certain macro-algae
(Lurencia, Fucus, Laminaria,
Macrocystis, Halimeda, and
Acetabularia)
3 The phytoplankton (see Figure 7)
4 The epiphytic micro-algae
5 The benthic micro-algae
G The upper limits of thermal tolerance for
two species of copepods from Chesapeake
Bay were found to be near the normal
temperature of the habitat during the
summer. The addition of chlorine to the
cooling water killed all copepods passing
through the system at temperatures below
the upper limits of thermal tolerance.
Preliminary observation, also, indicates
that heat stress may stimulate oysters to
accumulate copper (as other stressful
factors are known to do) without being a
direct killing agent.
Dry weight of total estuarine epifauna
production averaged 2. 8 times greater
in the discharge canal than in the intake
area over a 5 year period in another study.
-------
Water Temperature and Water Quality
VI SUMMARY
The various environmental factors cannot be
considered as isolated entities, organisms
respond to the entire environment. Tern"
perature criteria thus must be based on the
requirements of the entire aquatic population,
and on the life history requirements for
different seasons of the year.
REFERENCES
1 Berger, Bernard B. Does Production of
Power Pollute our Rivers' Power
Engineering. March 1961.
2 Cairnes.J. Jr. Effects of Increased
Temperature on Aquatic Organisms
Industrial Wastes. Vol. 1 (4):
150-152. 1956.
3 Clark, S.M. and Snyder, G. R. Timing
and Extent of a Flow Reversal in the
Lower Columbia River. Jour. Limn,
of Ocean. Vol. 14. November 1969.
6 FWPCA Presentations, ORSANCO
Engineering Committee, Seventieth
Meeting, Terrace Hilton Hotel,
Cincinnati, Ohio. September 10, 1969.
7 Heibrun, L. V. Heat Death, Scientific
American, pp. 70*75. April 1954.
8 Pennsylvania State of. Heated
Discharges..., Their Effect on Streams
Pub. No. 3, Div. San. Engr., Bur.
Environmental Health, Dept. of Health,
January 1962.
9 Trembley, F.J. Research Project on
Effects of Condenser Discharge Water
on Aquatic Life. Progress Report.
Inst, of Research. Lehigh Univ.
November 21, 1960.
10 Chesapeake Science. Proceedings of the
2nd Thermal Workshop of the U. S.
International Biological Program.
Solomons, Md. Vol. 10 (3-4). 1969.
4 FWPCA, Water Quality Criteria Section
HI, Fish, Other Aquatic Life, and
Wildlife. 1968.
5 FWPCA, Northwest Water Laboratory.
Industrial Waste Guide on Thermal
Pollution. Revised 1968.
This outline was prepared by H. W. Jackson,
Chief Biologist, National Training Center,
Water PrQgrams Operations, EPA, Clnofnati,
OH 45268.
-------
EFFECTS AND CONTROL OF OIL POLLUTION
I INTRODUCTION
The increased size of oil tankers, density
of waterborne traffic, and offshore petro-
leum production operations point up the
need for prevention and control of oil
spillage. For example, the Torrey Canyon
wreck dumped upwards of 30 million gallons
of oil on the coast of England in 1967, the
Ocean Eagle wreck spread over three
million gallons m San Juan Harbor alone.
Other major spills also occurred in the
York River, Virginia, and on the Cape Cod
National seashore in 1967, and the well-
known Santa Barbara, California, oil well
incident contaminated over 400 miles of
coastline.
A Specific Areas of Interest
1 On-scene control of gross leakage
2 Destruction or recovery of open sea
or open water oil slicks
3 Disposal of recovered mixtures
4 Protection of the shore face and
estuaries
5 Cleaning of shore face and estuaries
2 Floating Sorbers
a Inexpensive
b Can be disposed of by burning or
burial
c Oil recovery very difficult
3 Plastics (burning not recommended)
a Expensive
b Excellent solution to problem
since no residues are left on the
bottom
c Large quantities of oil can be
reclaimed for subsequent use
4 Gelling Agents
a In developmental stage
b May prove helpful in collecting
spilled oil
5 Sinkants
a Sink oil, such as by certain chalks
or carbonized sand
6 Effect of oil pollution and treatment
agents on marine and freshwater flora
and fauna
7 Waterfowl recovery methods
B Floating Sorbers and Sinkants
1 Examples of collecting agents
a Floating sorbers such as straw,
or mineral perlites
b Plastic or other polymeric
materials such as polyurethane
foam
c Gelling agents
b Demersal fish species may be
affected adversely
c Resurfacing of oil mass generally
is probable, although it is delayed
and slow
d Method should be applied only
beyond continental shelf and in
areas not involving commercial
fisheries
6 The spreading of sorbers, polymeric
materials, gelling agents, etc., is
aided when spread by specialized
equipment.
7 Confined slicks are easier to handle
WP.OI. 1.9. 72
-------
Effects and Control of Oil Pollution
C Chemical Treatment
1 A large number of dispersants and
emulsifiers exist, although little
quantitative or comparative informa-
tion on toxicities and effectiveness to
aquatic life is available.
2 The aromatic oil solvents used in the
compounding of many of these agents
have been found to be the principal
toxic agent.
3 Based on review of the Torrey Canyon
incident, opinion is divided on the
general use of oil emulsifiers when
water pollution control is of prime
consideration. *
D Biological Degradation of Crude Oil and
Oil Fractions in the Ocean and Fresh
Water
1 Biological degradation is controlled
by the following environmental
conditions.
a Nutrients
b Temperature
c O^ availability
d Degree of dispersion of oil
e Species of bacteria
f Other species of invertebrates
2 Degradation rates are very slow.
3 Seeding of oil slicks with micro-
organisms which feed specifically on
the oil requires further research.
E Booming
1 The ability to confine a spill is
principally a function of time,
availability and efficiency of equip-
ment, and prevailing environmental
conditions.
2 'If oil release occurs over several
days, the ability to confine the oil to
the immediate area would depend
largely on the prevailing water
conditions
a Booming in waters with a sea
state greater than three is
impractical with most of the
presently existing boom designs.
b Even with a sea state of three or
less, this is suspect if wind
conditions are adverse.
c Currents in excess of 1. 5 to 2
knots make booming difficult
without extensive skirts and
anchoring systems
3 There are two principal types of
mechanical barriers applicable to
oil spills--floating booms and under-
water bubble barriers.
a Both presently are suitable only
for calm water and are subject
to failure
b The floating boom is generally
superior to the bubble barrier
on an emergency basis as it is
more portable
c The main advantage of a bubble
barrier is the unrestricted entry
and exit of ships
*Review Dept. of Interior policy on use of chemicals to treat oil (in lack of national
contingency plan) available as handout, or from Dept. of Interior.
-------
Effects and Control of Oil Pollution
The main disadvantage is the
complete loss of containment m
the event of air supply failure
G Skimming
F Burning
If all attempts to salvage a vessel or
cargo have failed and a ship has been
abandoned, an attempt should be
made to set the oil afire while still
contained in the vessel.
Burning on the surface generally is
not effective due to the rapid transfer
of heat to the water, selective burning
of lighter-fractions, and lack of O^
supply except at the edge of a slick.
Caution should be exercised in the
vicinity of ships, docks, etc.
Mechanical devices for collecting
oil from the surface of water, such
as rotating cylinders and suction
pumps, generally are available but
have relatively small capacities
The use of these devices is restricted
to relatively,calm waters if reason-
able efficiency is to be achieved
This outline was prepared by John Wooley,
formerly Aquatic Biologist, Pacific Northwest
Water Laboratory, WPO, EPA, Corvallis,
OR 97330
-------
BIOTIC EFFECTS OF SOLIDS
I Sedimentation of rivers, lakes, estuaries
and adjacent coastal water should be con-
sidered as a special case of pollution
resulting from deforestation, overgrazing
and faulty agricultural practices, road
construction, and other land management
abuses.
A Good farming practices can do a great
deal to prevent silt from reaching streams
and lakes.
B Road building and housing development
projects, placer mining, strip mining,
coal and gravel washing, and unprotected
road cuts are important sources of
turbidity that can be reduced with planning,
good housekeeping, and regulation.
II Setteable solids include both inorganic and
organic materials which may settle out
rapidly, forming bottom deposits of both
inorganic and organic solids.
A They may adversely affect fisheries by
covering the bottom of the stream or lake
with a blanket of material that destroys
the bottom fauna or the spawning grounds
of fish (Figur' 1 from Ingram, et al).
NOTE
(SOUfiCE OF POLLimON FROM
GLASS-SANO WASTES)
OftAPM KEY
Figure 1 Vertical bar graph*, superimposed ever a map, used to show
total genera and individuals of bottom animals per unit area
B The organic fraction includes such
setteable materials as greases, oils,
tars, animal and vegetable fats, feed
lot wastes, paper mill fibers, synthetic
plastic fibers, sawdust, hair, greases
from tanneries, and various settleable
materials from city sewers. Deposits
containing organic materials may deplete
bottom oxygen supplies and produce
hydrogen sulfide, carbon dioxide, methane,
or other noxious gases.
C The inorganic components include sand,
silt, and clay originating from such
sources as erosion, placer mining, mine
tailing wastes, strip mining, gravel
washing, dusts from coal washeries,
loose soils from freshly plowed farm
lands, highway, and building projects.
D Some settleable solids may cause damage
by mechanical action.
E The biota of streams is limited by the
type of substrate.
1 A depositing substrate generally
contains fewer types and may be
dominated by burrowing forms.
2 An eroding substrate has a charac-
teristic fauna of sessile attached and
foraging members, such as bryozoans,
stoneflies, nonburrowmg mayflies,
and net-spinning caddis flies.
3 The addition of solids over an
originally eroding riffle substrate
brings about pronounced changes in
the biological community from diatoms
to fish. The following are common
macroinvertebrates of this new "trickling
filter" community in contrast to E 2 above.
a Oligochaetes
b Alderfly larvae (Sialis)
c Midge larvae (Chironomids)
WP. 13c. 9. 72
-------
Biotic Effects of Solids
III Turbidity, color, and transparency are
closely interrelated phenomena in water.
They must be observed simultaneously
because transparency is a function of tur-
bidity, water color, and spectral quality of
transmitted light.
A Turbidity is an expression of the optical
property of a sample of water which
causes light to be scattered and absorbed
rather than transmitted'in straight lines
through the sample.
B Turbidity is caused by the presence of
suspended matter such as clay, silt,
finely divided organic matter, bacteria,
plankton, and other microscopic
organisms.
1 Algae, turbidity from silts and clays,
and color of the water all affect one
environmental factor of major impor-
tance m the productivity of aquatic
wildlife habitat--light penetration of the
water.
a Excessive turbidity reduces light
penetration in the water and, there-
fore, reduces photosynthesis by
phytoplankton organisms, attached
algae, and submersed vegetation.
b The results of many of man's
activities, including agriculture,
industry, navigation, channelization,
dredging, land modification, and
eutrophication from sewage or
fertilizers, often reduce light trans-
mission to the degree that aquatic
angiosperms of value to wildlife
cannot grow.
. c Mixed effluents from various
industrial plants and domestic
sewage increase the turbidity of
receiving water. It is difficult to
distinguish between the effect of
the attenuation of light due to sus-
pended particles and the direct
effect of the particles in suspension
on the growth physiology of aquatic
organisms.
2 In many coastal waters, the principal
cause of turbidity is the discharge of
silt carried out by the principal rivers.
Secchi disc readings show that the
transparency of water at the mouths
of large rivers during flood stage may
be reduced to a few centimeters. At
normal river stages, the disc may be
visible at several meters below the
surface.
3 Dredging of bays and tidal rivers for
improvement of navigation occasionally
presents serious problems. Benthic
communities in the area near dredging
operations may be destroyed or
damaged by spoil deposition, mcrease
in water turbidity, release of toxic
substances accumulated in the mud
of the polluted areas, and by changing
the pattern in the dredged area.
IV PHYSICAL DAMAGES FROM
SILTATION
A Silt and sediment are particularly
damaging to gravel and rubble-type
bottoms. The sediment fills the inter-
stices between gravel and stones, thereby
eliminating the spawning grounds of fish
and the habitat of many aquatic insects
and other invertebrate animals such as
mollusks, crayfish, fresh water shrimp,
etc.
B Accumulation of silt deposits is de-
structive to marine plants, not only by
the associated turbidity, but by the
creation of a soft, semi-liquid substratum
inadequate for anchoring the roots.
Back Bay, Virginia and Currituck Sound,
North Carolina serve as examples of the
destructive nature of silt deposition.
Approximately 40 square miles of bottom
are covered with soft, semi-liquid silts
up to 5 inches deep; these areas, con-
stituting one-fifth of the total area,
produce only 1 percent of the total aquatic
plant production.
-------
Biotic Effects of Solids
V S^LT POLLUTION INCLUDES NOT ONLY
PURELY PHYSICAL EFFECTS, BUT
ALSO MAY INCLUDE COMPLEX MATERIAL.
A Pollution in the estuary may be derived
from contamination hundreds of miles
upstream in the river basin or it may be
of purely local origin. Silt plays a major
role in the transport of toxicants, especially
pesticides, down to the estuary.
1 Agricultural chemicals are adsorbed
on silt particles. Under poor farming
practices, as much as 11 tons of silt
per acre per year may be washed by
surface water into a drainage basin,
2 Surface mining and deforestation further
accelerate the process of erosion and
permit the transport of terrestrial
chemical deposits to the marine
environment.
B Oil that settles to the bottom of aquatic
habitats can blanket large areas and
destroy the plants and animals of value
of waterfowl.
1 Reportedly, some oil sludges on the
bottoms of aquatic habitats tend to
concentrate pesticides, thus creating
a double hazard to waterfowl that
would pick up these contaminants in
their normal feeding process.
2 Observations on storage of carcinogenic
compounds found in oil-polluted water
and on affected sediments are biolog-
ically significant, since they may be
concentrated by commercially
harvested bivalves.
C Much of the tonnage of aerially applied
pesticides fails to reach the designated
spray areas and the presence of 5 ng/1
of DDT in presumably untreated Alaskan
rivers indicates the magnitude of this
facet of the pollution problem.
1 The continuous presence of 5 fig/1 of
DDT in the marine environment would
decrease the growth of oyster populations
by nearly 50 percent.
2 Atmospheric drift is also an important
factor in the transport of a variety of
pollutants to ths aquatic environment.
3 Organochlorine compounds from sources
other than pesticides applications are
involved in food webs and biological
magnification m remote polar environ-
ments.
D ''the data on water pollution, however, are
less encouraging. Among other things, they
indicate that land runoff from farms and
even urban land, as opposed to discharges
from cities and factories, has a much greater
impact on water pollution than we realized.
In all type3 of river basins, the concentration
of nutrients, which can eutrophy our lakes,
is increasing. These data indicate that while
we carry on our major efforts to clean up
pollution from municipal and indjstrial
sources, we must increasingly turn our
attention to l^nd runoff-of nutrients, fertil-
izers, pesticides, organic materials, and
the soil particles that often transport the
others. If we fail to do so, our expenditures
for water quality will not achieve maximum
improvement. " Council on Environmental
Quality.
REFERENCES
1 Cairns, John. Suspended Solids Standards
for the Protection of Aquatic Orga-
nisms. Proc. 22, Ind. Waste Conf.,
Purdue Univ. Ext. Ser. 129.16. 1967.
2 FWPCA Southeast Water Lab. Role of
Soils and Sediment in Water Pollution
Control. Part I., Athens, Ga.
90 pp. 1968.
3 FWPCA Missouri Basin Region. Second
Compendium on Animal Waste
Management. USDA, Kansas City,
Mo. 256 pp. 1969.
4 Hall, James D. Alsea Watershed Study
(To determine the effects of logging
on aquatic resources). Dept. Fish &.
Wildlife. Oregon State Univ. 11 pp.
1967.
-------
Bio tic Effects of Solids
5 Isom, Billy G. The Mussel Resources
of the Tennessee River, Malacologia.
7(2-3):397-425. 1969.
6 Manheim, Frank T., Meade, Robert H.,
and Bond, Gerard C. Suspended
Matter in Surface Waters of the
Atlantic Continental Margin from Cape
Cod to the Florida Keys. Science
167(3917):371-376. 1970.
7 Weidner, R.B, Christianson, A.G , Weibel,
S. F. and Robeck, G. G. Rural Runoff as
a Factor m Stream Pollution. JWPCF.
41<3):377-384.
8 Patrick, Ruth. Effect of Suspended
Solids, Organic Matter and Toxic
Materials on Aquatic Life in Streams.
Water & Sewage Works. 115 89. 1968.
9 Council on Environmental Quality.
Environmental Quality, The Third
Annaal Report. Aug. 1972.
This outline was prepared by Ralph M. Sinclair,
Aquatic Biologist, National Training Center,
WPQ EPA, Cincinnati, OH 45268.
-------
GLOBAL, DETERIORATION AND OUR ENVIRONMENTAL CRISIS
I FROM LOCAL TO REGIONAL TO GLOBAL
PROBLEMS
A Environmental problems do not stop at
national frontiers, or ideological barriers.
Pollution in the atmosphere and oceans
taints all nations, even those benignly
favored by geography, climate, or natural
resources.
1 The smokestacks of one country can
pollute the air and water of another.
2 Toxic effluents poured into an inter-
national river can kill fish in a
neighboring nation and ultimately
pollute international seas.
B In Antarctica, thousands of miles from
pollution sources, penguins and fish
contain DDT in their fat. Recent layers
of snow and ice on the white continent
contain measurable amounts of lead.
The increase can be correlated with the
earliest days of lead smelting and com-
bustion of leaded gasolines.
C International cooperation, therefore, is
necessary on many environmental fronts.
1 Sudden accidents that chaotically
damage the environment - such as oil
spills from a tanker at sea - require
international cooperation both for
prevention and for cleanup.
2 Environmental effects cannot be
effectively treated by unilateral action.
3 The ocean can no longer be considered
a dump.
D "One of the penalties of an ecological
education is that one lives alone in a
world or wounds. Much of the damage
inflicted on land is quite invisible to
laymen. An ecologist must either harden
h\s shell and make believe that the conse-
quences of science are none of his business,
or he must be the doctor who sees the marks
of death in a community that believes
itself well and does not want to be told
otherwise.11 Aldo Leopold
E CHANGES IN ECOSYSTEMS ARE
OCCURRING CONTINUOUSLY
A Myriad interactions take place at every
moment of the day as plants and animals
respond to variations in their surroundings
and to each other. Evolution has produced
for each species, including man, a genetic
composition that limits how far that
species can go in adjusting to sudden
changes in its surroundings. But within
these limits the several thousand species
in an ecosystem, or for that matter, the
millions in the biosphere, continuously
adjust to outside stimuli. Since inter-
actions are so numerous, they form long
chains of reactions.
B Small changes in one part of an ecosystem
are likely to be felt and compensated for
eventually throughout the system.
Dramatic examples of change can be seen
where man has altered the course of
nature. It is vividly evident in his well-
intentioned but poorly thought out tampering
with river, lake, and other ecosystems.
1 The Aswan High Dam
2 The St. Lawrence Seaway
3 Lake Kariba
4 The Great Lakes
5 Valley of Mexico
6 California earthquake (Scientific
American 3981, p. 333)
7 Everglades and the Miami, Florida
Jetport
8 Copperhill, Tennessee (Copper Basin)
9 (You may add others)
Bl. ECO. hum. 2d. 9. 72
-------
Global Deterioration and Our Environmental Crisis
C Ecosystem Stability
1 The stability of a particular ecosystem
depends on its diversity. The more
inter dependencies in an ecosystem, the
greater the chances that it will be able
to compensate for changes imposed
upon it.
2 A cornfield or lawn has little natural
stability. If they are not constantly
and carefully cultivated, they will not
remain cornfields or lawns but will
soon be o/ergrown with a wide variety
of hardier plants constituting a more
stable ecosystem,
3 The chemical elements that make up
living systems also depend on complex,
diverse sources to prevent cyclic
shortages or over supply.
4 Similar diversity is essential for the
continued functioning of the cycle by
which atmospheric nitrogen is made
available to allow life to exist. This
cycle depends on a wide variety of
organisms, including soil bacteria and
fungi, which are often destroyed by
pesticides in the soil.
2 Invertebrates
a Asian Clams have a pelagic veliger
larvae, thus, a variety of hydro
installations are vulnerable to sub-
sequent pipe clogging by the adult
clams.
b Melanian snails are intermediate
hosts for various trematodes
parasitic on man.
3 Vertebrates
a At least 25 exotic species of fish
have been established in North
America.
b Birds, including starlings and
cattle egrets.
c Mammals, including nutria.
4 Aquatic plants
Over twenty common exotic species
are growing wild in the United States.
The problem of waterway clogging has
been especially severe in parts of the
Southeast.
5 Pathogens and Pests
D Biological Pollution
Contamination of living native biotas by
introduction of exotic life forms has been
called biological pollution by Lachner et al,
Some of these introductions are compared
to contamination as severe as a dangerous
chemical release. They also threaten to
replace known wildlife resources with
species of little or unknown value.
1 Tropical areas have especially been
vulnerable. Florida is referred to as
"a biological cesspool of introduced
life".
Introduction of insect pests and tree
pathogens have had severe economic
effects.
Ill LAWS OF ECOLOGY
A Four principles have been enunciated by
Dr. Barry Commoner.
1 Everything is connected to everything
else.
2 Everything must go somewhere.
3 Nature knows best.
4 There is no such thing as a free lunch.
B These may be summarized by the principle,
"you can't do just one thing. "
-------
Global Deterioration and Oar Environmental Crisis
IV THE THREE PRINCIPLES OF
ENVIRONMENTAL CONTROL (Wolman)
A You can't escape.
B You have to organize.
C You have to pay.
V POLLUTION COMES IN MANY PACKAGES
A The sources of air, water, and land
pollution are interrelated and often
interchangeable.
1 A single source may pollute the air
with smoke and chemicals, the land
with solid wastes, and a river or lake
with chemical and other wastes.
2 Control of air pollution may produce
more solid wastes, which then pollute
the land or water.
3 Control of wastewater effluent may
convert it into solid wastes, which
must be disposed of on land, or by
combustion to the air.
4 Some pollutants - chemicals, radiation,
pesticides - appear in all media.
B "Disposal" is as important and as costly
as purification.
VI PERSISTENT CHEMICALS IN THE
ENVIRONMENT
Increasingly complex manufacturing
processes, coupled with rising industrialization,
create greater amounts of exotic wastes
potentially toxic to humans and aquatic life.
They may also be teratogenic (toxicants
responsible for changes in the embryo with
resulting birth defects, ex., thalidomide),
mutagenic (insults which produce mutations,
ex., radiation), or carcinogenic (insults
which induce cancer, ex., benzopyrenes)
in effect.
A Metals - current levels of cadmium, lead,
and other substances whose effects on
humans and fish and wildlife are not fully
understood constitute a mounting concern.
Mercury pollution., for example, has
become a serious national problem, yet
mercury has been present on earth since
time immemorial. More research is
needed, yet we dare not relax our
standards until definitive answers have
been provided.
B Pesticides
1 A pesticide and its metabolites may
move through an ecosystem in many
ways. Hard (pesticides which are
persistent, having a long half-life in
the environment includes the organo-
chlorines, ex., DDT) pesticides
ingested or otherwise borne by the
target species will stay in the
environment, possibly to be recycled
Dr concentrated further through the
natural action of food chains if the
species is eaten. Most of the volume
of pesticides do not reach their target
ai all.
2 Biological magnification
Initially, low levels of persistent
pesticides in air, soil, and water
may be concentrated at every step
up the food chain. Minute aquatic
organisms and scavengers, which
screen water and bottom mud having
pesticide levels of a few parts per
billion, can accumulate levels
measured in parts per million -
a thousandfold increase. The sediments
including fecal deposits are continuously
recycled by the bottom animals.
a Oysters, for instance, will con-
centrate DDT 70, 000 times higher
in their tissues than it's concentration
in surrounding water. They can
also partially cleanse themselves
in water free of DDT.
-------
Global Deterioration and Our Environmental Crisis
b Fish feeding on lower organisms
bmld up concentrations in their
visceral fat which nay reach several
thousand parts per million and levels
in their edible flesh of hundreds of
parts per million.
c Larger animals, such as fish-eating
galls and other birds, can further
concentrate the chemicals. A sj-vey
on organochlorine residues in aquatic
birds m the Canadian prairie provinces
showed that California and ring-billed
gulls were among the most con-
taminated. Since gulls breed in
colonies, breeding population changes
can be detected and related to levels
of chemical contamination. Ecological
research on colonial birds to monitor
the effects of chemical pollution on
the environment is useful.
C "Polychlorinated oiphenyls" (PCB's).
PCB's are used in plasticizers, asphalt,
ink, paper, and a host of other products.
Action has been taken to curtail their
release to the environment, since their
effects are similar to hard pesticides.
D Other compounds which are toxic and
accumulate in the ecosystem:
1 Phalate esters - may interfere with
pesticide analyses
2 Benzophyrenes
3
great influence on previously unpolluted
waters and their life.
C Mimmata, Japan and mercury pollution.
D Organochlorine levels in commercial and
sport fishing stocks, ex., the lower
Mississippi River fish kills.
E Yoj may complete the following
1
2
VIH SUMMARY
A Ecosystems of the world are linked
together through biogeochemical cycles
which are determined by patterns of
transfer and concentrations of substances
in the biosphere and surface rocks.
B Organisms determine or strongly
influence chemical and physical charac-
teristics of the atmosphere, soil, and
waters.
C The inability of man to adequately predict
or control his effects on the environment
is indicated by his lack of knowledge
concerning the net effect of atmospheric
pollution on the earth's climate.
D Serious potential hazards for man which
are all globally dispersed, are radionuclides,
organic chemicals, pesticides, and
combustion products.
VII EXAMPLES OF SOME EARLY WARNING
SIGNALS THAT HAVE BEEN DETECTED
BUT FORGOTTEN, OR IGNORED.
A Magnetic micro-spherules in lake
sediments now used to detect changes
in industrialization mdjcate oar slowness
to recognize indicators of environmental
change.
B Salmonid fish kills in poorly buffered
clean lakes m Sweden. Over the pas+
years there had been a successive
increase of SO^ in the air and precipitation.
Thus, air-borne? contamination from
industrialized European countries had a
E Environmental destruction is in lock-step
with our population growth.
ACKNOWLEDGEMENT:
This outline has been extracted in part from
the first annual report of the Council on
Environmental Quality: Environmental
Quality. USGPO, Washington, DC.
326 pp? $1.75. 1970.
REFERENCES
1 Goldman, Charles R. Is +he Canary Dying'
The time has come for man, miner of
the worlds resources, to surface. Calif.
Medicine 113:21-26. 1970.
-------
2 Lachner, Ernest A. , RobinSj C. Richard,
and Courtenay, Walter R., Jr.
Exotic Fishes and Other Aquatic
Organisms Introduced into North
America. Smithsonian Contrib. to
Zool. 59:1-29. 1970.
3 Nnagu, Jerome O. and Bowser, Carl J.
The Magnetic Spherules in Sediments
of Lake Mendota, Wisconsin. Water
Res. 3 833-842. 1969.
4 Rhode, Wilhelm. Limnology, Social
Welfare, and Lake Kmneret. Ititernat.
Assoc. Theory and Appl. Limnology.
17-40-48. 1969.
5 Hood, Donald W. ed. Impingement of Man
on the Oceans. Wiley-Interscience.
738 p. 1971.
6 Commoner, Barry. The Closing Circle,
Nature, Man, and Technology. Alfred
A. Knopf. 326 p. 1971.
Glooal Deterioration and Our Environmental Crisis
12 Matthews, W. H , Smith, F. E., and
Goldberg, E. D. Man's Impact on
Terrestrial and Oceanic Ecosystems.
MIT Press. 1971.
13 Wagner, Richard H. Environment and
Man. W W Norton & Co., Inc.
NY. 491 p. 1971.
14 Leopold, Aldo. A Sand County Almanac
with Essays on Conservation from
Round River. Sierra Club/Ballantine
Books. 205 p. 1970.
15 Sondheimer, Ernest B. and Simeone,
John B. Chemical Ecology. Academic
Press. 336 p. 1970.
16 Whittaker, Robert H. Communities and
Ecosystems. Macmillan Co. 162 p.
1970.
17 Environmental Quality. Second Annual Report
of the Council on Environmental Quality.
August 1971. Third Annual Report.
August 1972.
18 Toxic Substances. Council on
Environmental Quality. 25 p.
April 1971.
19 Zinc m Water. A Bibliography USDI.
Office Water Resources WRSIC Series
208. 1971. Also in this series WRSIC
201-207, Mercury, Magnesium,
Manganese, Copper, Trace Elements,
and Strontium.
7 Dansereau, Pierre ed. Challenge for
Survival. Land, Air, and Water for
Man in Megalopolis, Columbia CJniv.
Press. 235 p. 1970.
8 Caudill, Harry M. My Land is Dying.
E. P. Dutton. 144 p. 1971.
9 Berkowitz, David A. and Squires,
Arthur M. editors. Power Generation
and Environmental Change. MIT Press,
441 p. 1971.
10 Wiens, John A. ed. Ecosystem Structure
and Function. Oregon State Univ. Press.
176 p. 1972.
11 Burns, Noel M. and Ross, Curtis editors.
Project Hypo, An Intensified Study of the
Lake Erie Central Basin Hypolimnion and
Related Surface Water Phenomena. 182 p.
CCIW, paper no. 6 and EPA Tech.
Report TS-05-71. 208-24. 1972.
This outline was prepared by Ralph M.
Sinclair, Aquatic Biologist, National
Training Center, Water Programs Operations,
EPA, Cincinnati, OH 45268.
-------
CONVERSION FACTORS
Length
1 centimeter
1 inch
1 meter
1 foot
1 meter
1 yard
1 Kilometer
I mile
0 394 inch
2 540 centimeters
3 2808 feet
0 305 meter
1 0936 yards
0 9144 meter
0 62137 mile
1 60935 kilometers
Area
1 square centimeter
1 square inch
1 square meter
1 square foot
1 square meter
1 square yard
1 bquurc kilometer
1 square mile
1 acre (US)
0 1550 square inch
6 452 square centimeters
10 764 square feet
0 09290 square meter
1 1960 square yards
0 8361 square meter
0 3861 square mile
2 590 square kilometers
4840 square yards
Volume
1 cubic centimeter
1 cubic inch
1 cubic meter
1 cubic foot
1 cubic meter
1 cubic yard
0 0610 cubic inch
16 3872 cubic centimeters
35 314 cubic feet
0 02832 cubic meter
1 3079 cubic yards
0 7646 cubic meter
Capacity
1 milliliter
1 ounce (U IS liquid)
1 milliliter
1 dram (U S Apothecaries)
1 liter
1 quart (U S liquid)
1 liter
1 gallon (U S liquid)
0 03382 ounce (U S liquid)
29 573 milliliters
0 2705 dram
(U S Apothecaries)
3 6967 milliliters
1 05671 quarts (US liquid)
0 94633 liter
0 26418 gallon (U S liquid)
.i 78533 liters
Mass
1 gram
1 grain
1 gram
1 ounce (Avoirdujjois)
1 gram
1 ounce (Troy)
1 kilogram
1 pound (Avoirdupois)
15 4324 grains
0 0648 gram
0 03527 ounce (Avoirdupois)
28 3495 grams
0 03215 ounce (Troy)
31 10348 grams
2 20462 pounds (Avoirdupois)
0 45359 kilogram
Power
1 watt
1 foot pound per second
I watt
I BTU per minute
I watt
1 Horsepower (US)
I watt
I kilogram-caloric per minute
1 watt
1 lumen
0 73756 foot pound per second
1 35582 watts
0 056884 BTU per minute
17 580 watts
0 001341-Horsepower (U.S )
745 7 watts
0 01433 kilogram-calorie per
minute
69 767 watts
1 x 10' ergs per second
0 001496 watt
DEGtaS
F.
210
200-
19i
160
"-F80
170
140"
150-
U0- -AO
c
100
¦90
¦ 70
130-
120-
110—
•40
101
»0-
80
-30
70"
-20
60-
50'
-10
40-
30-
- 0
20-
-10
-------
SECTION H
BIOLOGICAL METHODS AND TECHNIQUES
The concept that use of water Invariably results in changes in the quality or
characteristics of the water was covered in the preceding section.
Methods then must be developed whereby a given water resource can be
evaluated to determine its suitability for use and/or to determine the nature
and extent of changes in the water that have occurred through use. Data
interpretation demands a thorough knowledge of the conditions under which
the samples were collected and a critical assessment of the reliability of the
data's representation of the situation. Representative data sheets included
here are not intended as a standard, but are designed to show how field
observations may be integrated with subsequent laboratory analysis. Adequate
field notes are essential for correct interpretation of laboratory data.
In this section consideration is given to a number of biological techniques for
evaluation of water quality, and their application in action programs.
Contents of Section H
Outline No.
Fundamentals of the Toxicity Bioassay 37
Biomonitoring of Industrial Effluents 40
Biological Field Methods 41
Stream Invertebrate Drift 42
Artificial Substrates 43
Attached Growths (Periphyton or Aufwuchs) 44
Application of Biological Data 46
Procedures for Fish Kill Investigations 47
An Initiation into Statistics ST 51 48
Using Benthic Biota in Water Quality Evaluations 49
-------
FUNDAMENTALS OF THE TOXICITY BIOASSAY
I INTRODUCTION
A The toxicity bioassay procedure herein
discussed is intended for use by indus-
trial and other laboratories.
B Its objective is to evaluate the toxicity
of wastes and other water pollutants to
fish or other aquatic organisms.
C This basic procedure evaluates relatively
acute toxicity only (chronic or cumulative
toxicity requires more extensive study).
D Potential applications are numerous.
1 Dilution and/or treatment necessary
to avoid acute toxic effects can be
estimated.
2 The efficacy of a treatment can be
tested.
3 The potential usefulness of a proposed
treatment can be estimated.
E The toxicity bioassay technique does not
involve a chemical knowledge of the
toxicant.
1 Synergism, antagonism, and other
interactions of chemical components
cannot always be anticipated, but are
automatically included in the overall
evaluation.
2 All chemical and physical information
available is essential to the adequate
interpretation and application of test
results.
F The test is designed for local application.
Generalizations should be made with
great caution.
G Field observations should be made of
results of application over a significant
period of time.
H Careful distinction should be made
between fish mortality due to a phys-
iological toxicant, and that due to lack
of DO.
I A uniform testing procedure is essential
to effective action in water pollution
control.
II ROUTINE PROCEDURE
A Test animals should be fish or other
organisms of local significance.
1 Extremely resistant or extremely
sensitive species should not be
selected.
2 They should be species which are
amenable to captivity.
3 They should be accurately identified.
4 They should be relatively uniform in
size. Individuals less than 3 inches
in length are usually most convenient.
5 They should be healthy and thoroughly
acclimated to the laboratory.
B Test water should preferably be taken
from the receiving stream just above
the discharge being evaluated.
1 If this is unsuitable, cleaner waters
from an upstream station may be
substituted.
2 Artificial "standard" waters are not
recommended for general use.
C Other Experimental Conditions
BI. BIO. met. 7c. 10.69
-------
Fundamentals of the Toxicity Bioassay
1 Temperature
The tests should be performed at a
uniform temperature in the upper part
of the expected summer range, e.g.,
20° - 25°C for warm water fish, and
12° - 180 c for cold water species.
It has been found however, that for
most routine operations, ambient
laboratory temperatures are satisfactory.
Standard modern air conditioning,
particularly if it is maintained 24 hours
a day, is quite adequate.
2 Test containers should be of glass.
Wide mouthed pickle jugs" or battery
jars are satisfactory. Five and one
gallon sizes are both useful, but the
larger size is required for conclusive
results.
3 Artificial Aeration should not be used
to maintain the dissolved oxygen
concentration. If this falls below
approximately 4 or 5 ppm at any time
during the test, fewer fish should be
used per container or an auxiliary
oxygenation procedure invoked that is
designed to void undue loss of volatile
toxicants.
4 The number of test animals should not
be less than 10 per concentration for
reliable conclusions, these may be
distributed between two or more
containers.
5 Ratio of fish to solution
There should be less than 2 grams of
fish per liter of test solution,
preferably not more than one.
D Experimental Procedure
1 All dilutions for a given run should be
prepared from the same sample.
2 Control tests are essential.
3 Duration
Tests should be run for at least 48 hours,
preferably 96.
4 Dead fish should be removed as soon
as observed. Survivors should be
counted and recorded each 24 hours.
5 Feeding during the test should be
avoided.
6 Experimental concentrations
Any appropriate concentrations may
be used. A logarithmic series such
as is suggested in Table I is very
convenient. Concentrations can be
expressed in percent by volume,
parts per million by weight, or other
appropriate units.
7 Expression of results
The measure of relative toxicity is
the median tolerance limit (symbol:
TL , this is the analogue of the
LDj.0 of the toxicologist).
a This is the concentration which
just 50% of the test animals can
survive for a stipulated period of
time (sometimes written TL*
where t 3 24, 48, 96 hours, etc.)
b The TL* may conveniently be
estimate1? graphically, by plotting
the experimental data on semi-
logarithmic graph paper, with the
test concentration laid off on the
log, scale, and the percent
survival on the arithmetic scale.
Connect with a straight line the
two successive data points
representing survival values of
greater than and less than 50%.
Note the concentration which
corresponds to 50% survival on
this graph. This is the "TL^
m
Other methods are acceptable.
Ill REPORTING, INTERPRETATION AND
APPLICATION
A Reports should include an orderly
tabulation of all pertinent data such as-
1 Identity of experimental animals
-------
Fundamentals of the Toxicity Bioassay
Their source, average size and
condition, and number used per
concentration
Source and chemical and physical
analysis of experimental water
Experimental temperature
Volumes of experimental liquid in
each container
The problem is to extrapolate from
this well established mid concentration
to a safe concentration well below the
"critical concentration range".
Initiation of regulatory procedures
based on the TL should be followed
by periodic fielcPobservations. If
aquatic life flourishes, there is no
problem indicated. If not, the
material must be still further diluted.
6 Records of running analyses such as
DO and pH
TL and data from which it was
, m
determined
B Interpretation and application will be
discussed more thoroughly later.
Briefly
1 The TL is an estimate of the midpoint
of the critical concentration range
(the interval between the highest
concentration at which all test animals
survive, and the lowest at which they
all die).
TABLE
A Guide to the Selection of Expe
Based on Progressive Bisection
Logarithmic Scale.
IV SPECIAL PROBLEMS
A Unaerated aquaria with finite quantities
of toxicant are not always satisfactory
(Static Tests).
1 The toxicant may be volatile.
2 Toxic materials may be masked by
a high BOD.
3 The toxicant may be progressively
adsorbed or otherwise changed.
rimental Concentrations,
of Intervals on a
Col. 1 Col. 2 Col. 3 Col. 4 Col. 5
10.0
3. 2
5.6
1.8
7.5
4.2
2.4
1.35
8. 7
6.5
4. 9
3. 7
2. 8
2. 1
1.55
1. 15
1.0
-------
Fundamentals of the Toxicity Bioassay
B Standards or requirements other than
those involving toxicity per se may be
involved.
C Preliminary and Concurrent Investigations
1 Obtain all available information about
unknown to be tested.
2 Does the material lend itself to this
type of test9
3 Run feasible on the spot analyses
including DO.
4 Significant quantities of solutions
removed from test containers for
analysis should be replaced with
similar volume of same dilution.
D Wastes with a high BOD or COD
1 Suggested preliminary tests
a Set up two identical exploratory
tests.
b Aerate one but not the other.
c If great difference develops
between them, special procedures
are indicated.
2 Oxygenation or aeration of dilution
water before making dilutions may
help.
3 Oxygenation of experimental containers
during run. Pure oxygen is suggested
instead of air in order to avoid the
bubbling any more gas through the
containers than is necessary as some
of the toxic fraction may be volatile
materials which would be stripped out.
a Lead oxygen into tank through glass
tube instead of breaker stone in
order to keep bubbles large.
b Control rate of bubbling. Keep it
at the minimum number of
bubbles per minute which will
maintain 4 to 5 milligrams of
oxygen per liter. Do not attempt
saturation.
c Other systems of oxygenation are
available.
4 Renewal of solutions at stated inter-
vals (12, 24, or 48 hours) is approved.
Fish are not harmed by being
carefully transferred from one
container to another. It is useful
where-
a Initial DO is adequate but slowly
exhausted.
b Toxicant is volatile, progressively
adsorbed, precipitated, or other-
wise changed.
E Continuous flow apparatus is highly
desirable but expensive.
1 Equipment more involved and subject
to failure during a run.
2 May be adapted to monitoring by use
of proportioning equipment. Makes
longer runs possible.
F Other Considerations
1 Radioactive wastes must be evaluated
in regard to their chemical toxicity
as well as their radioactivity.
2 Sub acute levels of many toxicants
such as lead, arsenic, chromium,
etc., may exert a low level chronic
toxicity over a long period of time.
3 "Safe levels" of a waste in regard to
toxicity may still exceed standards
of other types such as color, organic
content, suspended solids, etc.
-------
Fundamentals of the Toxicity Bioassay
REFERENCES
1 American Public Health Association,
** Standard Methods for the Examination
of Water and Wastewater, 12th edition.
New York. 1965.
2 Doudoroff, P., et al, Bio-Assay
Methods for the Evaluation of Acute
Toxicity of Industrial Wastes to Fish,
Sew. and Ind. Wastes, Vol. 23, No. 11.
November 1951.
3 Doudoroff, P. arid Katz, M. Critical
Review of Literature on the Toxicity
of Industrial Wastes and Their
Components to Fish. I. Alkalies,
Acids and Inorganic Gases, Sew, and
Ind. Wastes, Vol. 22, No. 11, 1432.
November 1950.
4 Ellis, M.M., Westfall, B.A. and
Ellis, M.D. Determination of
Water Quality, Research Report 9,
U. S. Fish and Wildlife Service,
122 pp. 1946.
5 Hart, W.B., Doudoroff, P. and
Greenbank, J. The Evaluation of the
Toxicity of Industrial Wastes,
Chemicals and Other Substances to
Fresh-Water Fishes. The Atlantic
Refining Company, Philadelphia, Pa.
317 pp.
6 Hart, W.B., Weston, R.F. and
DeMann, J. F. An Apparatus for
Oxygenating Test Solutions in Which
Fish are Used as Test Animals for
Evaluating Toxicity. Trans. Am.
Fisheries Soc. 1945, 75, 228 pub.
1948.
This outline was prepared by H. W. Jackson,
Chief Biologist, National Training Center,
Water Programs Operations, EPA,
Cincinnati, OH 45268.
-------
BIOMONITORING OF INDUSTRIAL EFFLUENTS
I INTRODUCTION
A Plant operating personnel need to know
the general quality of an effluent which is
being discharged at a fairly constant rate
and also must be warned if a slug of toxic
material is released to the receiving
water.
B Conventional bioassay procedures can
evaluate only single samples taken at
particular times. Continuous-flow bio-
assays of single grab samples over a long
period of time can be very useful, but do
not solve the problem of transient
variations.
A To demonstrate the continuous suitability
of an effluent for aquatic life provided
slow-acting or cumulative toxins are not
involved. The continuous testing of an
undiluted effluent, (Objective A) is usually
accomplished by leading a small stream
of the effluent through an aquarium. This
aquarium may be located in a public lobby
to enhance public relations, or it may be
in the plant for operational use only. This
is a relatively simple and direct approach
and needs no further elaboration.
B To detect change (usually deleterious) in
the biological acceptability of the effluent
itself.
C A technique that does permit exercising
continuous surveillance over the toxicity
of an effluent is biomomtoring
(2 3 4)
D Conventional bioassays ' ' can provide
important information about the actual
toxicity of batches of the effluent in terms
of TL 's, and, if sufficient samples are
tested about the range of variation. Bio-
assays should be run from time to time to
ascertain the exact toxicity of a waste
even though it is being monitored as outlined
below.
E The following procedures refer only to
toxic wastes having a relatively rapid
action. Wastes such as cadmium which
have long delaycumulative effects at low
concentrations, oxygen-demanding
wastes, radioactive wastes, and others,
would either be inappropriate or would
not elicit a recognizable reaction soon
enough to be of use.
C To detect change in the effect of the
effluent on the biota of the receiving water.
Ill EQUIPMENT
A single basic design of exposure tanks and
flow plan can be used to accomplish Objective
B or C. (Figure 1) With the exception of a
simple suggestion for proportioning flow of
effluent to dilution water (Figure 2), engineer-
ing devices for accomplishing the varioug ^ g.
needs outlined below are not discussed. ' '
Special care should be used to ensure that all
surfaces that come in contact with the waste
or the dilution water are constructed of non-
toxic and noncorrosive materials. This pre-
caution is particularly necessary for marine
waters, where bimetallic contact s are very
dangerous. An experienced aquatic biologist
should be consulted in the preparation of
plans.
A Exposure Tanks
II OBJECTIVES OF BIOMONITORING
Three basic objectives of biomonitoring
are-
1 Exposure tanks (Figure 1) should be
large enough (10 to 20 gallons) that the
test organisms c^ live normally under
plant conditions. Simple construction
BI. BIO. 26. 3. 67
-------
Biomomtoring of Industrial Effluents
CONSTANT
SOURCE 01-
EFFLUENT
CONSTANT
SOURCE OF
DILUTION WATER
REGULATING DEVICE
hi
t
— k
EXPOSURE
TANK NO.l
PROPORTIONING
¥
DEVICE
EXPOSURE
TANK NO. 2
PROPORTIONING
DEVICE
EXPOSURE
TANK NO. 3
SCREENED
OVERFLOW
OUTLETS
Figure 1. SCHEMATIC FLOW PLAN FOR OBJECTIVES B OR C
-------
Biomomtoring of Industrial Effluents
EFFLUENT DILUTION WATER
HEAD r -v
Figure 2
SUGGESTED PLAN FOR PROPORTIONING FLOW BETWEEN EFFLUENT
AND DILUTION WATER. (See 1 and j. Figure 1)
-------
Biomonitormg of Industrial Effluents
will facilitate feeding, cleaning, and
disease control. "Eye-appeal" is not
necessary unless public relation is
involved, but scrupulous sanitation is
essential as in all long-term animal
culture. Tanks should be
situated in a lighted and well-ventilated
room, but not exposed to direct
sunlight. Ambient room temperatures
are generally satisfactory.
2 Inlet and overflow should be similar in
all tanks so that conditions will be the
same except for the quality of the water.
The total flow of water through the tank
should be adjusted so that the hydraulic
retention times are equal, whether the
total flow is coming from one source
or two.
3 Tank No. 1 contains unadulterated
dilution water to establish that the
test animals will live in it. This tank
is the control or "reference" to which
the other tanks are compared.
4 Tanks 2 and 3 (more may be added
at points 1) contain mixtures of
effluent and dilution water. If the
experimental animals die or show dis-
tress in these tanks, a change for the
worse in the characteristics of the
effluent being monitored is probably
indicated.
5 l and j are mixing or proportioning
devices set to predetermined amounts.
6 All tanks overflow to the sewer through
opening k, which should be screened
to prevent escape of test animals and
clogging in outlet pipe.
B Effluent Supply
The supply of effluent should be constant
and controllable. The prime requisite
is that it be fresh so that changes may be
detected at the earliest possible moment.
C Dilution Water
1 The source and quality of the dilution
water is the most important single
factor m the system because this is
the scale or standard by which the
toxicity of the effluent is assayed.
2 To test the effect of the effluent on
the receiving water (Objective C), the
dilution water obviously must be the
receiving water.
a In most cases the receiving water
varies from time to time as runoff
water washes in different materials
from the surface of the land the tide
turns, or other plants release
various wastes.
b Plan C (Objective C) automatically
evaluates the toxicity of the effluent
when discharged into this changing
situation. This water should con-
tain all of the components present
down to the outfall being monitored,
but none of the effluent itself.
3 In a stream situation, dilution water
can be taken well upstream from the
waste discharge. A pipeline with a
continuous flow is ideal, since slugs
of material from upstream sources
that might modify the toxicity of the
effluent being monitored would be
taken into the monitoring system and
quickly distributed to the exposure
tanks. Because such systems are
notoriously subject to "problems, "
batch transportation of control water
may have to be employed. In lakes,
estuarine, or coastal situations, batch
supply may be the only practical
solution.
4 If the receiving water is already
continuously toxic to aquatic life, it is
unsuitable for use in the system.
a Under such circumstances Plan C
would be unworkable and the only
recourse would be Plan A or B.
-------
Biomonitoring of Industrial Effluents
b Treated water such as drinking
water from a city or industrial plant
should never be used for any of
these plans, even if dechlorinated,
because the chemicals used in treat-
ment may react with the waste being
monitored.
D Proportioning Flow
The plan (Figure 2) by which effluent and
dilution water are proportioned to the test
tanks (Tank 2 and 3) determines what the
system will accomplish.
1 A suggested plan for proportioning flow
between effluent and dilution water
(i and j) in Figure 1 is given in Figure
2.
2 Rubber tubes (n) can be momentarily
diverted to catch flow in an appropriate
sampling device such as a graduated
cylinder. Based on the time required
to discharge some standard volume,
flow could be proportioned to any de-
sired ratio (for example, one part
effluent to two parts dilution water).
3 The removal of sediment by some sort
of trap as shown is a policy matter
that should be resolved in the project-
design statement.
IV FLOW PLANS
A Flow Plan for Objective B
If the system is to be operated to detect
only serious detrimental change in the
effluent itself, Tanks 2 and 3 should con-
tain mixtures in such proportions of
effluent and dilution water as to permit
the test animals to live as long as the
effluent is normal.
1 One possible combination would be to
adjust the mixing mechanism at i
(Figure 1) to admit such a proportion
of effluent that the test animals in Tank
No. 1 could barely survive. The
slightest increase in toxicity of the
effluent would then immediately be
made evident by the death of the test
animals and appropriate remedial
action could be taken.
2 The mixing mechanism j in Tank No. 3
might be adjusted to provide a greater
margin of safety, for example 1/2 or
1/ 10th the toxicity of Tank 2. If Tank
2 animals then died, but Tank 3 animals
survived, it would presumably indicate
only a moderate increase in toxicity.
B Flow Plan for Objective C
Monitoring the effect of the effluent on
the receiving water, from the point of
view of protecting aquatic life, is ideal,
but may also be very difficult.
1 The reference tank (e, Figure 1) re-
ceives water fresh from the receiving
body. This water is free of any trace
of the effluent being monitored, but
contains all substances, natural and
artificial, presently in the receiving
water. These components may change
from time to time, and one of these
changes may increase the toxic effect
of the effluent (synergism).
Thus the death of animals in the
strongest test tank, but not in the
reference tank, may be the result of
an increase in the toxicity of the plant
waste, or of a synergistic reaction of
the effluent with a material in the
receiving water.
2 No matter the cause, the death itself
serves as a warning and immediate
action can be taken on the discharge to
protect aquatic life.
3 On the other hand, if a slug of strongly
toxic matter enters the receiving water
from some outside source, the deaths
of the animals in the reference Tank 1
(and probably also those in exposure
Tanks 2 and 3) would show that the
"fish kill" presumably in progress in
nature was not the result of the effluent.
-------
Biomonitoring of Industrial Effluents
4 A parallel installation under Plan B
might also demonstrate no change m
the waste being monitored.
5 Numerous other dilution systems might
also be employed under Plan C.
C A different approach is to proportion the
mixture to simulate the actual mixture
taking place in the receiving water. For
this purpose if the location were on a
flowing stream, the flow of both the stream
and final effluent must be known. Periodic
adjustments might be made by hand, or by
automatic equipment involving telemetry
of both effluent and stream flow.
V TEST ANIMALS AND OTHER
CONSIDERATIONS
No universal recommendations can be made
about test animals to be used although many
suggestions are available. 3, 5)
A Irwin^11) investigated the suitability of
57 species of freshwater fishes for this
purpose. Briefly, they should be of local
importance, they must be a type that can
be maintained in good health in the labora-
tory in the dilution water to be used, and
enough must be employed so that
reasonable statistical reliability is
assured (for example, 10 per tank).
1 Fish are usually employed as test
animals, although there is no reason
not to use any other organisms that
can be successfully kept alive in the
test tanks.
2 Continuous availability is also impor-
tant. It is well not to change the test
species after a program has been
established, since the reactions of
different species to the waste being
monitored may not be the same. (H)
3 Animals in the exposure tanks should
be fed the same as those in the stock
tanks. The same kind of food in the
same ratio of food to weight of test
fish should be added to each tank at
the same intervals. Unnatural accep-
tance of food in exposure tank may in-
dicate a measure of distress, even in
the absence of mortality.
B Oxygen determinations should be run
occasionally to ensure that any deleterious
symptoms are the result of the effluent
and not of oxygen deficiency. Actual
minimum acceptable levels will depend
on the temperature and type of fish used.
C Long-continued exposure to low level
concentrations in tanks may result in
cumulative intoxication, or acclimatization.
1 In most cases these effects can probably
be best counteracted by periodic re-
newal of the test fishes, for example-
at 60-day intervals.
2 When obtaining stocks of test animals
from receiving waters, these same
factors should be borne in mind. Fish
or other organisms taken from below
the outfall might have acquired some
immunity or sensitivity to the effluent
being tested.
3 Fish or a species normally present
in the receiving stream, but imported
from some other (unpolluted) source,
will presumably exhibit a completely
unconditioned response. Generally,
since the aquatic life to be protected
is that already present in the stream,
the most logical source of fish is the
stream itself. Under operating condi-
tions, however, it is not always
practicable to collect the experimental
animals from this source and imports
from another area may be necessary.
D Selection of Dilutions
1 The "critical range" of toxicity may
be defined as the range between the
highest concentration that kills no
test animals and the lowest concentra-
tion that kills all. The TLm of the
conventional bioassay(3) is m the middle
portion of this range.
-------
Biomomtormg of Industrial Effluents
When an effluent is to be biomonitored
under Plan B, the selection of appro-
priate dilutions might be based on the
above concept of "critical range. "
a If a "tight" control is desired, the
highest concentration might be es-
tablished near the TLm. When a
batch of test animals is first placed
in such a dilution, approximately
half of them may (by design) be
expected to die. The survivors,
however, would constitute a
rigorous control as any increase in
toxicity would be expected to kill
the remaining animals m order of
susceptibility, until, as the top of
the critical range is reached, all
would be dead.
b A somewhat less stringent control
would be effected with a dilution
near the lower end of the critical
range.
In any large population of test animals
kept in an exposure tank over an ex-
tended period of time, an occasional
animal may be expected to die.
a The mortality of significance then
is not the occasional individual
death, but the sudden death of 25,
50, or 100% of the test animals.
b When this happens, biomomtormg
has sounded the alarm to take
appropriate action to detoxify the
effluent or to divert it from the
receiving stream until it is again
normal.
2 American Public Health Association, Inc.
Standard Methods for the Examination
of Water and Wastewater. Part VI.
Bioassay Methods for the Evaluation of
Acute Toxicity of Industrial Waste-
waters and other Substances to Fish.
Twelfth Edition. New York. 1965.
3 Weiss, Charles M. Use of Fish to Detect
Organic Insecticides in Water. Journal
Water Pol. Cont. Fed. 37(5) -647-658.
1965.
4 Henderson, Croswell and Tarzwell,
Clarence M. Bio-Assays for the Con-
trol of Industrial Effluents. Sewage
and Ind. Wastes 29(9) -1002-1017.
1957.
5 Pickering, Quentin H. Research in Pro-
gress. 1966.
6 Mount, Donald I. and Warner, Richard E.
A Serial-Dilution Apparatus for the
Continuous Delivery of Various Con-
centrations of Material in Water. U.
S. Dept. of HEW, PHS Publication
No. 999-WP-23. June 1965.
7 Clark, John R. and Clark, R. L. Sea-
Water Systems for Experimental Aqua-
riums - A Collection of Papers. USDI.
Research Report 63. 1964.
8 Symons, James M. Simple, Continuous-
Flow Low and Variable Rate Pump.
Jour. Water Pol. Cont. Fed., 35(11):
1480-1485. 1963.
9 Emmens, C.W. Keeping and Breeding
Aquarium Fishes. Academic Press,
Inc. N. Y. 1953.
REFERENCES
1 Henderson, Croswell and Pickering,
Quentin, H. Use of Fish in the Detec-
tion of Contaminants in Water Supplies.
Jour. AWWA, 55(6) 715-720. 1963.
10 Lewis, W. M. Maintaining Fishes for
Experimental and Instructional Purposes.
Southern 111. Univ. Press, Carbon-
dale, 111.
-------
Biomonitoring of Industrial Effluents
11 Irwin, William H. Fifty-seven Species
of Fish m Oil Refinery Waste Bioassay.
Trans. Thirtieth N. Am. Wild, and
Nat. Res. Conf. March 8 - 10, 1965.
Wild. Mgmt. Inst. Wire Bid. Washing-
ton, D. C.
12 Klock, John W. and Pearson, Erman A,
Engineering Evaluation and Development
of Bioassay Kinetics. State Water Pol.
Cont. Board, Sacramento, Calif.
September 1961.
13 Mount, Donald I.
January 1966.
Personal Communication.
This outline was prepared by H W Jackson,
Chief Biologist, National Training Center
Water Programs Operations, EPA, Cincinnati, OH
45268 and W A Brungs, Jr , Aquatic
Biologist, Fish Toxicology, Newtown, OH 45244
-------
BIOLOGICAL FIELD METHODS
I INTRODUCTION
A Due to the nature of ecological inter-
relationships, methods for the collection
of different types of aquatic organisms
differ. In general we can recognize
those that swim or float and those that
crawl, those that are big and those that
are little. Each comprises a part of
"the life" at any given survey station
and consequently a "complete" collection
would include all types.
B Field methods in the following outline
are grouped under four general
categories, the collection of-
1 Benthos (or bottom dwelling
organisms). These may be
attached, crawling, or burrowing
forms.
2 Plankton (plancton). These are all
of the microscopic plants and
animals normally swimming or
suspended in the open water
3 Periphyton or "aufwuchs". This is
the community of organisms
associated with the surfaces of
objects. Some are attached, some
crawl. The group is intermediate
between the benthos and the plankton
4 Nekton. Nekton are the larger,
free swimming active animals such
as shrimp or fishes.
C Aquatic mammals and birds, in most
cases, require still other approaches
and are not included.
D There is little basic difference between
biological methods for oceanic,
estuarine, or freshwater situations
except those dictated by the physical
nature of the environments and the
relative sizes of the organisms.
Fish, benthos, and plankton collection
is essentially the same whether con-
ducted in Lake Michigan, Jones'
Beach, or the Sargasso Sea.
1 Marine organisms range to larger
sizes, and the corrosive nature of
seawater dictates special care in
the design and maintenance of
marine equipment. Site selection
and collection schedules are
influenced by such factors as tidal
currents and periodicity, and
salinity distribution, rather than
(river) currents, riffles, and pools.
2 Freshwater organisms are in
general smaller, and the water is
seldom chemically corrosive on
equipment. Site selection in
streams involves riffles, falls,
pools, etc., and a unidirectional
flow pattern. Lake collection may
involve less predictable strati-
fication or flow patterns.
E Definite objectives should be established
in advance as to the size range of
organisms to be collected and counted,
i.e. microscopic only, microscopic
and macroscopic, those retained by
"30 mesh" screens, invertebrates and/
or vertebrates, etc.
n STANDARD PROCEDURES
A Certain standard supplementary
procedures are a part of all field
techniques. In order to be interpreted
and used, every collection must be
associated with a record of environ-
mental conditions at the time of
collection.
1 Data recorded should include the
following as far as practicable.
Location (name of river, lake, etc.)
BI.MET.fm. Id.3.71
-------
Biological Field Methods
Station number (particular location
of -which a full description should
be on record)
Date and hour
Air temperature
Water temperature (at various
depths, if applicable)
Salinity (at various depths, if
applicable)
Tidal flow (ebb or flood)
Turbidity (or light penetration, etc.)
Weather
Wind direction and velocity
Sky or cloud cover
Water color
Depth
Type of bottom
Type of collecting device and
accessories
Method of collecting
Type of sample (quantitative or
qualitative)
Number of samples at each station
Chemical and physical data, e.g.,
dissolved oxygen, nutrients, pH,
etc.
Collector's name
Miscellaneous observations (often
very important)
2 All collecting containers should be
identified at least with location,
station number, sample number,
and date. Spares are very handy.
43-2
3 Much transcription of data can be
eliminated by using sheets or cards
with a uniform arrangement for
including the above data. The
same field data sheet may include
field or laboratory analysis.
B Compact kits of field collecting equip-
ment and materials greatly increase
collecting efficiency, especially if
collection site is remote from
transportation.
III PERSONA L OBSERVA TION A ND
PHOTOGRAPHY *
A Direct or indirect observation of under-
water conditions has become relatively
efficient.
1 Diving spheres, pioneered by
William Beebe, Cousteau, Honot,
Willm, and Manad are proving
very important for deep water
observations.
2 Use of the aqualung permits direct
personal study down to over
200 feet.
3 Underwater television (introduced
by the British Admiralty for
military purposes) is now generally
available for biological and other
observations.
4 Underwater photography is
improving in quality and facility.
5 Underwater swimming or use of
SCUBA is quite valuable for direct
observation and collecting.
IV COLLECTION OF BOTTOM OR
BENTHIC ORGANISMS
A Shoreline or Wading Depth Collecting
Plates I, II
1 Hand picking of small forms
attached to or crawling on rocks,
-------
Biological Field Methods
BOTTOM GRABS
closed
Ekraan
Jackson
Shipek
PLATE I
-------
Biological Field Methods
LIMNOLOGICAL EQUIPMENT
j'-:
Hand Screen
Surber Sampler
Apron net
Specimen or
reagent bottles
Sorting pan
Pall
-------
Biological Field Methods
water is a fundamental and much
used method for quickly assaying
what is present and -what may be
expected on further search.
2 Patches of seaweed and eelgrass
and shallow weedy margins any-
where are usually studied on a
qualitative basis only.
a The apron net is one of the best
tools for animals in weed beds
or other heavy vegetation. It
is essentially a pointed wire
sieve on a long handle with
coarse screening over the top
to keep out leaves and sticks.
b Grapple hooks or a rake may
be used to pull masses of
vegetation out on the bank
where the fauna may be
examined and collected as they
crawl out.
c Quantitative estimates of both
plants and animals can be made
with a "stove pipe" sampler
which is forced down through
a weed mass in shallow water
and embedded in the bottom.
Entire contents can then be
bailed out into a sieve and
sorted.
d A frame of known dimensions
may be placed over an area to
be sampled and the material
within cropped out. This is
especially good for larger
plants and large bivalves
This method yields quantitative
data.
3 Sand and mud flats in estuaries and
shallow lakes may be sampled
quantitatively by marking off a
desired area and either digging
away surrounding material or
excavating the desired material
to a measured depth. Handle-
operated samplers recently
developed by Jackson and
Larrimore, make for more
effective sampling of a variety
of bottoms down to the depth of
the handles. Such samples are
then washed through graded
screens to retrieve the organisms.
4 Ekman grabs are most useful on
soft bottoms. This is a completely
closing clamshell type grab with
spring operated jaws. Size of grab
is usually 6" X 6" or 9" X 9", the
12" X12" size is impractical due
to its heavy weight when filled with
bottom material.
For use in shallow water, it is
convenient to rig an Ekman with
a handle and a hand operated jaw-
release mechanism.
5 The Petersen type grab (described
below) without weights will take
satisfactory samples in firm muds,
but tends to bury itself in very
soft bottoms. It is seldom used in
shallow water except as noted
below.
B Collecting in Freshwater Riffles or
Rapids
1 The riffle is one of the most
satisfactory habitats for comparing
stream conditions at different
points.
2 The hand screen is the simplest
and easiest device to use in this
situation. Resulting collections
are qualitative only.
a In use the screen is firmly
planted in the stream bed.
Upstream bottom is thoroughly
disturbed with the feet, or
worked over by hand by
another person. Organisms
dislodged are carried down
into the screen.
b Screen is then lifted and
dumped into sorting tray or
collecting jar.
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Biological Field Methods
3 The well-known square foot Surber
sampler is one of the best quan-
titative collecting devices for
riffles.
a It consists of a frame one foot
square with a conical net
attached. It is usable only in
moving water.
b In use it is firmly planted on
the bottom. The bottom stones
and gravel within the square
frame are then carefully gone
over by hand to ensure that all
organisms have been dislodged
and carried by the current into
the net. A stiff vegetable
brush is often useful in this
regard.
c From three to five square-foot
samples should be taken at each
station to insure that a reason-
able percentage of the species
present will be represented.
4 The Petersen type grab may be used
in deep swift riffles or where the
Surber is unsuitable.
a It is planted by hand on the
bottom, and worked down into
the bottom with the feet.
b It is then closed and lifted by
pulling on the rope in the usual
manner,
5 A strong medium weight dipnet is
the closest approach to a universal
collecting t6ol (effective for every-
thing from Blondes to indicator
organisms1)
a This is used with a sweeping
motion, through weeds, over
the bottoms or in open water.
A triangular shape is preferred
by some. This may be used
as a roughly quantitative device
in riffles by holding the end
flat against the bottom and
and backing slowly up-stream
disturbing the substrate with
one's feet. A standard period
of time is used.
b The handle should be from 4
to 6 feet long, and about the
weight of a garden rake
handle.
c The ring should be made of
steel or spring brass, and
securely fastened to the
handle. It should be strong
but not cumbersome; size of
ring stock will depend on
diameter of ring.
d The bag or net should be the
strongest available, not over
1/8 inch mesh, preferably
about 1/16 inch. Avoid 30 or
more meshes to the inch, this
is so fine that the net plugs too
easily and is slow and heavy
to handle.
e There should be a wide canvas
apron sewed around the rim
and protecting the bag. The
rim may be protected with
leather if desired.
D Deep Water Benthic Collecting Plate III
1 When sampling from vessels, a
crane and winch, either hand or
power operated, is used. The
general ideas described for shallow
waters apply also to deeper waters,
when practicable.
2 The Petersen type grab, seems to
be the best all around sampler for
the greatest variety of bottoms at
all depths, from shoreline down to
over 10, 000 meters. (Plate I)
a It consists of two heavily
constructed half cylinders
closed together by a strong
lever action.
-------
Biological Field Methods
DEEP WATER EQUIPMENT
Bathythermograph
Biological dredge
Otter trawl
PLATE III
-------
Biological Field Methods
b To enable them to bite into
hard bottoms, or to be used in
strong currents, weights may
be attached to bring the total
weight up to between 50 and
100 lbs.
c Areas sampled range from
l/5th to 1 /20th square meters
(1/10 square meter equals
approximately 1. 1 square ft.)
d A Petersen grab to be hauled
by hand should be fitted with
5/8 or 3/4 inch diameter twisted
rope in order to provide
adequate hand grip. It is best
handled by means of wire ropes
and a winch.
3 Other bottom samplers include the
VanVeen, Lee, Holme, Smith-
Mclntyre, Knudsen, Ponar, and
others,
4 A spring loaded sampler has
recently been developed by Shipek
for use on all types of bottoms.
It takes a half-cylinder sample,
1 /25th square meters in area and
approximately 4 inches deep at the
center. The device is automatically
triggered on contact with the
bottom, and the sample is com-
pletely protected enroute to the
surface. (Plate I)
5 Drag dredges or scrapes are often
used in marine waters and deeper
lakes and streams, and comprise
the basic equipment of several types
of commercial fisheries. Some
types have been developed for
shallow streams. In general
however, they have been little used
in fresh water.
6 The above is only a partial listing
of the many sampling devices
available. Others that are often
encountered are the orange-peel
bucket, plow dredge, scallop type
dredge, hydraulic dredges, and
various coring devices. Each has
its own advantages and dis-
advantages and it is up to the
worker and his operation to decide
what is best for his particular needs.
The Petersen type and Ekman grabs
are perhaps the most commonly
used.
7 Traps of many types are used for
various benthic organisms,
especially crabs and lobsters.
Artificial substrates (below) are in
essence a type of trap.
8 Since most biological communities
are not evenly distributed, it is
advisable to routinely take at least
two and preferably more samples
from any one station.
E Artificial substrates rely on the
ecological predilection of organisms
to grow wherever they fmd a suitable
habitat. When a small portion of
artificial habitat is provided, it tends
to become populated by all available
species partial to that type of situation.
The collector can then at will remove
the habitat or trap to his laboratory and
study the population at leisure.
This versatile research technique is
much used for both routine monitoring
and exploratory studies of pollution.
It is also exploited commercially,
especially for shellfish production.
Types of materials used include.
1 Cement plates and panels.
2 Wood (especially for burrowing
forms).
3 Glass slides (ex Catherwood
diatometer).
4 Multiple plate trap (masonite).
5 Baskets (or other containers) holding
natural bottom material and either
imbedded in the bottom, or sus-
pended m the overlying water.
-------
Biological Field Methods
6 Unadorned ropes suspended in the
water, or sticks thrust into the
bottom.
F Sorting and Preservation of Collections
1 Benthic collections usually consist
of a great mass of mud and other
debris among which the organisms
are hidden. Various procedures
may be followed to separate the
organisms,
a The organisms may be picked
out on the spot by hand or the
entire mess taken into the
laboratory where it can be
examined more efficiently
(especially m rough weather).
Roughly equivalent time will
probably be required in either
case.
b Specimens may be simply
observed and recorded or they
may be preserved as a
permanent record.
3
c Organisms may be simply
counted, weighed, or measured
volumetncally; or they may be
separated and recorded in
groups or species.
2 If separation is in the field, this is
usually done by hand picking,
screening, or some type of flotation
process.
a Hand picking is best done on a
white enameled tray using
light touch limnological forceps.
b Screening is one of the most
practical methods to separate
organisms from debris in the
field. Some prefer to use a
single fine screen, others
prefer a series of 2 or 3
screens of graded sizes. The
collection may be dumped
directly on the screen and the
mud and debris washed through,
or it may be dumped into a
bucket or small tub. Water
is then added, the mixture is
well stirred, and the super-
natant poured through the
screen. The residue is then
examined for heavy forms that
will not float up.
c A variation of this method in
situations where there is no
mud is to pour a strong sugar
or salt solution over the
collection in the bucket, stir
it well, and again pour the
supernatant through the screen.
This time, however, saving
the flotation solution for
re-use. The heavier-than-
water solution accentuates the
separation of organisms from
the debris (except for the
heavy shelled molluscs, etc.).
A solution of 2-1/2 lbs. of
sugar per gallon of water is
considered to be optimum.
Preservation or stabilization is
usually necessary in the field.
a 95% ethanol (ethyl alcohol) is
highly satisfactory. A final
strength of 70% is necessary
for prolonged storage. If the
collection is drained of water
and flooded with 95% ethanol
in the field, a laboratory
flotation separation can usually
be made later, thus saving
much time. Considerable
quantities of ethanol are
required for this procedure.
b Formaldehyde is more widely
available and is effective in
concentrations of 3 - 10% of
the commercial formulation
However, it shrinks and
hardens specimens, collector,
and laboratory analyst without
favor' In order to minimize
bad effects from formalin,
neutralized formalin is
-------
Biological Field Methods
recommended Mollusc shells VI
will eventually disintegrate in
acid formalin
A
c Properly preserved benthos
samples may be retained
indefinitely, thereby enhancing
their utility.
d Refrigeration or icing is very
helpful.
V MICROFAUNA AND PERIPHYTON
(OR AUFWUCHS) SAMPLING
A This is a relatively new area which
promises to be of great importance
The microfauna of mud and sand
bottoms may be studied to some extent
from collections made with the various
devices mentioned above In most
cases however, there is considerable
loss of the smaller forms
B Most special microfauna samplers for
soft bottoms are essentially modified
core samplers in which an effort is
made to bring up an undisturbed portion
of the bottom along with the immediately
overlying water. The best type currently
seems to be the Enequist sampler which
weighs some 35 kg. and takes a 100 sq
cm sample 50 cm deep.
B
C Microfauna from the surface of hard
sand or gravel bottoms may be sampled
by the Hunt vacuum sampler. This has
a bell-shaped "sampling" tube sealed
by glass diaphragm On contact with
the bottom, the glass is automatically
broken and the nearly bottom material
is swept up into a trap.
D Periphyton attached to or associated
with hard surfaces such as rock or
wood may be sampled by scraping or
otherwise removing all surface
material from a measured area. The
periphyton, however, is more effectively
quantitatively sampled by artificial
substrate techniques described above.
THE COLLECTION, OR SAMPLING
OF PLANKTON PLATE IV
Phytoplankton A Planned Program is
Desirable
1 A planned program of plankton
analysis should involve periodic
sampling at weekly* or even more
frequent intervals.
2 A well-planned study or analysis
of the growth pattern of plankton
in one year will provide a basis
for predicting conditions the
following year since seasonal
growth patterns tend to repeat
themselves from year to year.
a Since the seasons and the years
differ, records accumulated
over the years become more
useful.
b As the time for an anticipated
bloom of some troublesome
species approaches, the
frequency of analyses may be
increased.
3 Detection of a bloom in its early
stages will facilitate more
economical control
Field Aspects of the Analysis Program
1 Two general aspects of plankton
analysis are commonly recognized-
quantitative and qualitative.
a Qualitative examination tells
what is present.
b Quantitative tells how much.
c Either approach is useful, a
combination is best.
2 Equipment for collecting samples
in the field is varied.
a A half-liter bottle will serve
for surface samples of
phytoplankton, if carefully
taken.
-------
Biological Field Methods
PLANKTON SAMPLERS
Wisconsin net
High speed plankton sampler
PLATE IV
-------
Biological Field Methods
b A Kemmerer, Nansen, or
other special sampler (small
battery operated pumps are
time saving) is suggested for
depth samples.
c Plankton nets concentrate the
sample in the act of collecting
and also capture certain larger
forms which escape from the
bottles. Only the more
elaborate types are quantitative
however. For phytoplankton,
#20 or #25 size nets are
commonly used. Usually a net
diameter of 5 - 10 inches is
sufficient. The smaller forms
however, are lost through any
net.
C Zooplankton Collecting
1 Since zooplankton have the ability
to swim away from water bottles,
etc. nets towed at moderately fast
speed are used for their capture.
Number 12 nets (aperature
size 0. 119 mm, 125 meshes 1 inch)
or smaller numbered net sizes are
commonly used. A net diameter
greater than 5" is preferred.
Frequently half meter nets or
larger are employed. These may
be equipped with flow measuring
devices for measuring the amount
of water entering the net.
2 Other instruments such as the
Clark-Bumpus, Gulf-Stream,
Hardy continuous plankton recorder,
and high-speed instruments are
used for collecting zooplankton, also.
3 The devices used for collecting
plankton capture both the plant and
animal types. The mesh size
(net no.) is a method for selecting
which category of plankton is to be
collected.
D The Location of Sampling Points
1 Both shallow and deep samples are
suggested.
a "Shallow" samples should be
taken at a depth of 6 inches to
one foot. The surface film is
often significant.
b "Deep" samples should be
taken at such intervals
between surface and bottom
as circumstances dictate.
In general, the entire water
column should be sampled as
completely as practicable,
and the plankton from each
level recorded separately.
2 For estuarine plankton, it is
necessary to sample different
periods in the stage of the tide,
otherwise samples would be biased
to a given time, or type of water
carried by the tidal currents.
3 Plankton is subjected to the force
of the winds and currents. Asa
result, the plankton is often in
patches or "wind rows". For this
reason when using a net, it is often
desirable to tow the net at right
angles to the wind or current.
4 Not only are all plankton likely to
be horizontally discontinuous, but
zooplankton especially tend to be
numerous near the bottom in
daylight, but distributed more
evenly through the water column
at night. Therefore, a series of
tows or samples at different depths
is necessary to obtain a complete
sampling. One technique often
employed is to take an oblique tow
from the bottom to the top of the
water column.
5 Pilot studies to indicate sampling
locations and intervals are often
mandatory. Some studies require
random sampling points.
6 The number of sampling stations
that should be established is
limited by the capability of the
laboratory to analyze the samples,
but should approach the needs of the
objectives as closely as possible.
-------
Biological Field Methods
7 Field conditions greatly affect the
plankton, and a record thereof
should be carefully identified with
the collection as in II above.
8 Provisions should be made for the
field stabilization of the sample
until the laboratory examination
can be made if more than an hour
or so is to elapse.
a Refrigeration or icing is very
helpful, but ice should never
be placed in the sample.
b Preservation by 5% formalin is
widely used but badly shrinks
animals and makes all forms
brittle.
c Lugols solution is a good
preservative.
d Ultra-violet sterilization is
sometimes used in the laboratory
to retard the decomposition of
plankton.
e A highly satisfactory merthiolate
preservative has been developed
by the FWPCA and described by
Weber (1968).
VII COLLECTING FISH AND OTHER
NEKTON PLATES V, VI
A Fish and other nekton must be sought in
the obscure and unlikely areas as well
as the obvious locations in order for the
collection to be complete. Several
techniques should be employed where-
ever possible (this is appropriate for
all biota). It is advisable to check with
local authorities to inform them of the
reasons for samplmg, because many of
ttye techniques are not legal for the
layman. In this area, perhaps more
than any other, professionally trained
workers are important. Also, there
must be at least one helper, as a single
individual always has difficulty in pulling
both ends of a 20 foot seine simultaneously1
The more common techniques are
listed below.
B Seines
1 Straight seines range from 4-6 feet
and upwards m length. "Common
sense" minnow seines with approxi-
mately 1/4 inch mesh are widely
used along shore for collecting the
smaller fishes.
2 Bag seines have an extra trap or
bag tied in the middle which helps
trap and hold fish when seining in
difficult situations.
C Gill nets are of use in offshore and/or
deep waters. They range in length
from approximately 30 yards upward.
A mesh size is designed to catch a
specified size of fish. The trammel
net is a variation of the gill net.
D Traps range from small wire boxes or
cylinders with inverted cone entrances
to semi-permanent weirs a half mile or
more in length. All tend to induce fish
to swim into an inner chamber pro-
tected by an inverted cone or V - shaped
notch to prevent escape. Current
operated rotating fish traps are also
very effective (and equally illegal) in
suitable situations.
E Trawls are submarine nets, usually of
considerable size, towed by vessels at
speeds sufficient to overtake and scoop
in fish, etc. The mouth of the net must
be held open by some device such as a
long beam (beam trawl) or two or more
vanes or "otter boards" (otter trawl).
Plate III
1 Beam and otter trawls are usually
fished on the bottom, but otter
trawls when suitably rigged are
now being used to fish mid-depths.
2 The midwater trawl resembles a
huge plankton net many feet in
diameter. It is proving very effec-
tive for collecting at mid-depths.
-------
Biological Field Methods
FISH NETS
Gill
Hoop
Pound
PLATE V
-------
Biological Field Methods
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PLATE VI
-------
Biological Field Methods
Numerous special designs have been
developed. Plate VI
F Electric seines and screens are widely
employed by fishery workers in small
and difficult streams. They may also
be used in shallow water like areas with
certain reservations.
G Poisoning is much used in fishery studies
and management. Most widely used and
generally satisfactory is rotenone m
varying formulations, although many
others have been employed from time to
time, and some appear to be very good.
Under suitable circumstances, fish may
even be killed selectively according to
species.
H Personal observation by competent
personnel, and also informal inquiries
and discussions with local residents
will often yield information of real use.
Many laymen are keen observers,
although they do not always understand
what they are seeing. The organized
creel census technique yields data on
what and how many fish are being
caught.
I Angling remains in its own right a very
good technique in the hands of the skilled
practitioner, for determining what fish
are present. Spear-fishing also is now
being used in some studies.
J Fish and other nekton are often tagged
to trace their movements during
migration and at other times. Minia-
ture radio transmitters can now be
attached or fed to fish (and other
organisms) which enable them to be
tracked over considerable distances.
Physiological information is often
obtained in this way. This is known as
telemetry.
VIII SPECIA L REQUIREMENTS ON BOATS
Handling biological collections (as con-
trasted to chemical and physical sampling)
on board boats differs with the size of the
craft and the magnitude of operations.
Some possible items are listed below.
Hoisting and many other types of gear are
used in common with other types of
collection, and will not be listed.
A Special Laboratory Room(s)
B Constant flow of Clean water for
culturing organisms. (Selection of
materials and design of a system to
insure non-toxic water may be very
troublesome but very important.)
C Live Box built into ship at water level
D Refrigeration System(s)
1 For controlling temperature of
experimental organisms in
laboratory.
2 For deep-freezing and storage of
specimens to be examined later.
E Storage Space (Unrefrigerated)
F Facilities for the safe storage and use
of microscopes and other laboratory
equipment.
G Facilities for the safe storage and use
of deck equipment.
H Administrative access to the Captain
and Technical Leader in order to
coordinate requirements for biological
collection (such as a slow plankton tow)
with those for other collections.
I Safety of personnel working in and
around boats, as well as in other field
activities should be seriously con-
sidered and promoted at all times.
IX OTHER TYPES OF 'BIOLOGICAL
FIELD STUDIES INCLUDE
A Productivity Studies of Many Types
B Life Cycle and Management
C Distribution of Sport or (potentially)
Commercial Species
-------
Biological Field Methods
D Scattering Layers and Other Submarine
Sound Studies
E Artificial Culture of Marine Food Crops
F Radioactive Uptake
G Growth of Surface-Fouling Organisms
H Marine Borers
I Dangerous Marine Organisms
J Red Tides
K Others
X SOURCES OF COLLECTING
EQUIPMENT
Many specialized items of biological
collecting equipment are not available
from the usual laboratory supply houses.
Consequently, the American Society of
Limnology and Oceanography has compiled
a list of companies handling such items
and released it as "Special Publication
No. 1, Sources of Limnological and
Oceanographic Apparatus and Supplies. "
Available from the Secretary of the Society.
REFERENCES
1 Arnold, E.L., Jr. and Gehringer, J.W.
High Speed Plankton Samplers,
U.S. Fish and Wildlife Spec. Sci.
Rept. Fish No. 88:1-6.
2 Barnes, H. (ecj.). Symposium on New
Advances in Underwater Observations.
Brit. Assoc. Adv. Sci., Liverpool,
pp. 49-64. 1953.
3 Hedgepeth, Joel W. Obtaining
Ecological Data m the Sea Chapter 4
in "Treatise on Marine Ecology and
Paleoecology" Memois 67. Geol.
Soc. Am. 1963.
4 Isaacs, John D. and Columbus, O. D.
Oceanographic Instrumentation NCR
Div. Phys. Sci. Publ. 309, 233 pp.
1954.
5 Jackson, H.W. A Controlled Depth
Volumetric Bottom Sampler. Prog.
Fish Cult., April, 1970.
6 Lagler, Karl F. Freshwater Fishery.
Biology, Wm. C. Brown Company.
Dubuque. 1956.
7 Larrimore, R.W. Two Shallow Water
Bottom Samplers, (in press) 1969.
8 Morgan, A.H. Field Book of Ponds
and Streams. G.P. Putnam Sons.
New York. 1930.
9 Pennak, R.W. Freshwater Inverte-
brates of the United States. The
Ronald Press Company. New York.
1953.
10 Standard Methods for the Examination
of Water and Wastewater. (12th ed.
in print, 13th ed. in press, due in
print 1970). APHA, AWWA, WPFC.
Publ. by Am. Pub. Health Assoc.
New York.
11 Sverdrup, H.U. et al. Observations
and Collections at Sea, Chapter X
in The Oceans, Their Physics,
Chemistry, and Biology. Prentice-
Hall, Inc., New York. 1087 pp. 1942.
12 Usinger, R. L. Aquatic Insects of
California (Section on Field Methods).
University of California Press.
Berkeley. 1956.
13 Weber, C.I. The Preservation of
Phytoplankton Grab Samples.
Trans. Am. Mic. Soc. 87 70. 1968.
14 Welch, Paul S. Limnological Methods.
The Blakiston Company, Philadelphia,
Pennsylvania. 1948.
15 FWPCA, Investigating Fish Mortalities.
USDI, No. CWT-5, 1970.U.S. Gov't.
Print. Off. 1970 0-380-257.
This outline was prepared by H.W. Jackson,
Chief Biologist, National Training Center,
Water Programs Operations, EPA, Cincinnati,
OH 45268.
-------
STREAM INVERTEBRATE DRIFT
I Invertebrates which are part of the benthos,
but under certain conditions become carried
downstream in appreciable numbers, are
known as drift.
A Groups which have members forming a
conspicuous part of the drift include the
insect orders Ephemeroptera,
Trichoptera, Plecoptera and the crus-
tacean order Amphipoda.
B Other invertebrate groups exhibit drift
patterns.
II THREE BASIC TYPES OF DRIFT
ARE RECOGNIZED
A Catastrophic Drift
Floods wash numerous benthic organisms
downstream. Application of pesticides
may also cause such drift.
B Constant Drift (Incidental or Adventitious)
Organisms are constantly being dislodged
from the substrate during normal
activities and carried downstream.
C Periodic (Diel) or Behavioral Drift
In contrast to the other categories, this
is a specific behavior pattern and related
to circadian activity rhythms.
1 Seasonal drift occurs, for example,
in some maturing stoneflies which
drift downstream for emergence.
This is another reason for a serious
consideration of drift in bottom fauna
sampling since such presence of
stoneflies could easily be misinterpreted.
2 Periodic or diel drift occurs in peaks
for successive 24-hour periods.
a Night-active. Light intensity is the
phase-setting mechanism.
b Day-active. Water temperature
appears to be the phase-setter.
t
IE DIEL DRIFT
A Diel activity rhythms generally include
two peaks during the 24-hour period,
one major and the other minor.
1 The bigeminus type in which the
major peak occurs first (after sunset).
£
FIGURE 1
2 The alternans pattern with the major
peak occurring last.
x:
1200
1800
2400
0600
Time
Sunset Sunrise
FIGURE 2
B Drift Rate and Density (Waters, 1969)
1 Drift rate defined is ".. .the quantity
of organisms passing a width transect
or portion thereof, per unit time,
BI. ECO. 22a. 9.72
-------
Stream Invertebrate Drift
it is a measure of displacement or
the movement of organisms from one
place to another. "
2 Drift density .. is the quantity of
organisms per unit volume of water,
in much the same way as plankton
density can be defined. "
IV IMPLICATIONS FOR BIOLOGICAL
SAMPLING
A The drift from productive upstream
reaches may support a fish population
existing in relatively barren stream
sections.
B Drift will colonize artificial substrates,
such as suspended rock baskets, when
placed in such habitats.
C A bottom sampler such as the Surber,
could also be sampling drift when only
resident benthic organisms are intended
to be collected. This would depend on
the hour of collection and length of time
the Surber sampler is in the water.
D Application of drift studies have been
widely used in pesticide related studies.
In conjunction with such studies, Dimond
concluded that-
1 The status of drift is a much better
indicator of the steady state and of
total productivity than is the status
of the bottom fauna.
2 Bottom sampling, however, is
superior when analyzing survival
and recovery of the quality of population.
3 A combination of both in such a
sampling program would be most
likely to yield the most useful data.
E Drift sampling techniques have been
useful for recovery of large numbers of
sand-dwelling mayflies, which were
once rarely collected.
V MAJOR TAXA INVOLVED IN DRIFT
A The crustacean order Amphipoda
1 Gammarus species
2 Hyalella azteca
B The Insect Orders
1 Ephemeroptera
Baetis species (apparently universal)
2 Plecoptera
3 Trichoptera
4 Diptera
Simuliidae
5 Elmidae
C The main groups exhibiting very high
drift rates include- Baetis, some
Gammarus species, and some Simuliidae.
REFERENCES
1 Anderson, N. H. Biology and Down-
stream Drift of some Oregon
Trichoptera. Can. Entom. 99:507-
521. 1967.
2 Dimond, John B. Pesticides and
Stream Insects. Bull. 23, Maine
Forest Service, 21 pp. 1967.
3 Dimond, John B. Evidence that drift of
Stream Benthos is Density Related.
Ecology 48:855-857. 1967.
4 Pearson, William D., and Kramer,
Robert H. A Drift Sampler driven
by a Waterwheel. Limnology and
Oceanography 14(3) 462-465.
5 Reed, Roger J. Some Effects of DDT on
the Ecology of Salmon Streams in
Southeastern Alaska. Spec. Sci.
Report-Fisheries 542 1-15. U.S.
Bureau Comm. Fisheries. 1966.
-------
Stream Invertebrate Drift
6 Waters, Thomas F. Interpretation of
Invertebrate Drift in Streams.
Ecology 46 (3) 327-334. 1965.
7 Waters, Thomas F. Diurnal Periodicity
in the Drift of a Day-active Stream
Invertebrate. Ecology 49-152. 1968.
8 Waters, Thomas F. Invertebrate
Drift-Ecology and Significance to
Stream Fishes. (T.G. Northco e,
Ed.) Symposium Salmon and Trout
in Streams. University of British
Columbia, Vancouver. pp. 121-134.
1969.
This outline was prepared by R. M.
Sinclair, Aquatic Biologist, National
Training Center, Water Programs Operations,
EPA, Cincinnati, OH 45268.
-------
ARTIFICIAL SUBSTRATES
I INTRODUCTION THE NATURE OF
ARTIFICIAL SUBSTRATES
A Artificial substrates are anything
deliberately placed in the water for the
purpose of providing a place for benthic
or attached (sessile, sedentary, etc.)
organisms to grow on or in. This is in
contrast to "bait" which is used as an
attractant.
B Their origins for commercial use, or human
food production are rooted in antiquity.
Some examples are
1 Ropes, poles, brush, concrete
structures, and other objects thrust
into the bottom, or suspended in
estuarine waters to catch and grow
oysters and mussels (cultural techniques),
known virtually around the world.
2 Straw or reed tepees planted in shallow
alkaline lakes (in Mexico for example)
to catch the eggs of Corixids (Insecta.
Order Hemiptera, back-swimmers).
Eggs are harvested by drying and
brushing them off onto white sheets.
Used for human food.
C The fouling of ships bottoms, piling, etc.
by barnacles and other marine life is an
"artificial substrate in reverse".
D The use of aggregate to support a zoogloeal
mass of micro-biota in a trickling filter,
thus simulating a riffle area in a surface
stream, is a modern concept to harness
and make use of "consumer" and "reducer"
elements of a community in order to
dissipate the energy (oxidize, exhaust the
food value) contained in sewage.
II ECOLOGICAL BASIS
A Artificial substrates are based on the
"laws of organismal distribution. "
1 Any given kind of organisms tends to be
present (inhabit) in all available suitable
habitats.
*A community which has achieved a point of no further change, under a given set of
environmental conditions. Time scale may vary with circumstances.
NOTE- Mention of commercial products and manufacturers does not imply endorsement
by the Environmental Protection Agency.
2 Any given habitat tends to be inhabited
by all suitably adapted kinds of
organisms.
B A "substrate" being an object (or group
of objects) constitutes a habitat suitable
for sessile or attached organisms, and
also those that naturally burrow in, crawl
over, or otherwise live associated with
objects. Natural objects here could mean
the bottom, stones, sticks (floating or
sunk), etc.
C Organisms that would not be attracted to
substrates would be plankton and nekton
(fish and larger swimming invertebrates).
D Ecological Succession
Colonization is rapid in a biologically
productive water, and normally reaches
a stable climax* community in about a
month. A typical outline of successive
forms to appear in a freshwater situation,
for example, might be as follows.
1 Periphyton (slime forming) stage
(see also below)
a Bacteria - within an hour
b Diatoms - within the first day
c Other micro-algae - within the first day
d Protozoa - within the first day
2 Macroinvertebrate dominated stage
(see also below)
a Primary attached or sedentary
colonizers - second to third day
1) Net caddisflies
2) Bryozoa
3) Cordylophora caspia
4) Hydra
BI. MET. fm. 7d.9. 72
-------
Artificial Substrates
b Primary foragers
1) Mayflies
2) Stoneflies
3) Midges
c Secondary attached or sedentary
colonizers.
1) Sponges
2) Filamentous algae
d Adventitious forms
1) Crustaceans
2) Flatworms
3) Leeches
4) Snails
5) Other
3 Artificial substrates in a marine
environment proceed through similar
stages, except that the macroinvertebrate
stage may be more subject to variation
in the attachment of broods of barnacle,
oyster, and other larvae resulting from
greater numbers of types present, tidal
current variation, meteorological
conditions, etc.
Ill ARTIFICIA L SUBSTRATES AS SCIENTIFIC
COLLECTING DEVICES
A A review of the history of artificial
substrates for collecting microorganisms
(aufwuchs) (Cooke, 1956) indicates that
glass microscope slides were first used
for this purpose about 1915. Wood or
metal panels appear, however, to have been
deliberately exposed for the scientific
collection of larger organisms at least
since approximately the turn of the century,
and probably long before that (Visscher, 1928).
B Biological Applications
The principles of the artificial substrate
remain the same, regardless of the
community sampled. Two general types
of communities and associated samplers
have been employed
1 Periphyton (or aufwuchs) samplers
Periphyton is the community of slime
forming microorganisms which is the
first to attach to objects newly exposed
under water. This community is
generally considered to provide an
anchor layer to which other higher
forms of life can more readily attach.
It tends to persist until overgrown or
displaced by larger organisms, and
then in turn can be found spreading over
the surfaces of these same larger
plants and animals.
2 Periphyton has been widely studied as
it appears on 1 X3 glass microscope
slides which are equally convenient to
expose in the field and to study in the
laboratory.
3 Particular studies have included
a The original bacterial and fungal
slime
b Diatom identification and counts
c Identification and counts of other
microscopic algae
d Protozoans
e Primary productivity
4 The macroinvertebrate community is
sampled by a great variety of devices
such as those cited below. The
organisms are usually removed from
the substrate for study. Applications
have included the following:
a General study of the macroinverte-
brate community
-------
Artificial Substrates
b Estimates of productivity
c Studies of the life cycle of particular
species
d Studies of the influence of the sub-
strate on the attachment of sessile
forms
1) The influence of toxic paints for
the prevention of fouling organisms
2) Wood panels to study the pene-
tration of boring molluscs and
crustaceans
C Effect of type of device on what is collected
1 Wood boring organisms like teredo
worms (Mollusca, Pelecypoda) or
gribbles (Arthropoda, Isopoda) would
obviously be attracted primarily to
wood (although some are known to bore
in other materials).
2 Delicate forms and crawling forms
would be most likely to be collected on
devices having a shape to protect
against strong currents
3 Those with strong attachments could
endure swift currents, often, surpris-
mgly, even during periods of original
attachment (ex. byssus attached clams
which are also benthic forms).
4 Bottom burrowers would be most likely
collected in artificially contained
portions of bottom material.
D Effect of Location
1 The depth at which a sampler is sus-
pended may influence the organisms
attracted.
2 Location in or out of a current, direct
sunlight, etc , will influence the take.
E Some Types of Devices
1 Cement plates, panels, and blocks
2 Ceramic tiles
3 Wood blocks
4 Metal plates
5 Glass slides -1X3 inch micro slides
are used by many workers. Numerous
devices are employed to hold them.
They are generally either floated
(Weber and Raschke 1966) or sus-
pended in racks, anchored to
submerged bricks or other objects.
6 Plastic petri dishes
Burbanck and Spoon utilized an
ordinary 50 X 12 mm plastic petri
dish for collecting sessile protozoa,
Sickle modified this by using a
styrofoam cup (6 oz. size) with the
bottom third being cut off. The
lower unit of the plastic dish is
easily wedged into place in the cup
and the device is simply held by a
nylon line on a rope held in place by
an appropriate anchor and float.
The cup which tends to float is so
held that the petri dish bottom is in
a horizontal position and bottom side
up.
7 Multiple plate (Hester and Dendy, 1962)
a Common current procedure
utilizes 3-inch squares of 1/4
inch thick Masonite separated
by 1-inch square spacers.
These may be
b Threaded on an eye bolt or long
rod.
c Suspended by a loop of nylon cord.
8 Baskets or trays of bottom-type
material
a Trays of bottom material sunk in
the surface layer of the bottom.
b Baskets of stones suspended in
the water (Anderson and Mason,
1966).
9 Boxes, cages, bundles, etc., of
brush, reeds, or artificial material.
-------
Artificial Substrates
10 Polyethylene tapes
11 Plastic webbing
Minnesota Mining and Manufacturing
Company conservation web no. 200.
12 Styrofoam
13 Glass coverslips
Small slips are floated on the surface
of the water. Highly useful for protozoa
and rotifers. Remove and place on a
micro slide. Examine as a wet mount.
F Retrieval is an acute problem with all of
these samplers.
1 Physical factors
a Relocation
b Floods and drift
c High water
2 Well marked samplers or floats are
naturally vulnerable to the public,
resulting in disturbed, damaged, or
destroyed sample gear.
a This has been overcome by an
ingenious submerged float and
recovery line device. The weak
link in a submerged recovery line
is a modified flash bulb. An
electronic device actuated by an
underwater gun breaks the bulb
allowing the float and attached
line to surface. (Ziebell,
McConnell, and Baldwin)
b This unit has been further modified
by Fox (University of Georgia
Cooperative Fishery Unit) who
used an inexpensive detonator,
"Seal Salute". The latter is an
inexpensive fused charge designed
for underwater explosion.
IV ARTIFICIAL SUBSTRATES OR SAMPLERS,
AND WATER QUALITY
A Artificial substrates provide a habitat
("place to live"). It follows from the
laws of distribution (II A I and 2 above),
that the community which inhabits a
device will be governed by the physical
nature or structure interacting with the
characteristics of the surrounding water
(velocity, temperature, chemical
characteristics, etc.). Since the nature
of the sampler is controlled, it is evident
that the characteristics of the water
constitute the variable factor.
B Water Quality Surveillance
1 Similar substrates suspended side by
side in the same water tend to accumulate
(essentially) the same communities and
quantities of organisms.
2 Similar substrates suspended in different
waters accumulate different communities
and quantities.
3 Ergo* different communities and
quantities collected from similar
substrates at different places and times,
probably indicate different water qualities.
a These may be natural (seasonal,
diurnal, etc.)
b Or they may be a result of human
influences (pollution)
c A series of samplers the length of a
stream, lake, or estuary can suggest
"steady state" differences in water
quality.
d A series of samplers exposed over a
period of time at a given site can
suggest changes of water quality in
time.
4 The artificial substrate thus essentially
constitutes an in-situ bioassay of the
water.
-------
Artificial Substrates
C Interpretation and Significance of
Collections
1 The unit of comparison is most
appropriately taken as "the sampler".
The artificial substrate by definition is
not the natural local bottom material,
and unless it consists of a portion of
that bottom which has been actually
removed and replaced in an artificial
container (III-D-7)the composition and
magnitude of the community it contains
may or may not bear a definitive
relationship to the actual natural
problem The take of the artificial
substrate thus may have relatively
little relationship to the take of a
Petersen or an Ekman grab (dredge).
2 Comparisons between different types
of samplers are likewise hazardous
Each is what it is, and if they are
different they are not identical, thus
the biota each collects cannot be
expected to be identical (CF II A)
3 Artificial substrates should generally
be compared on a "sampler vs sampler"
basis, or for periphyton, "unit area
vs unit area".
REFERENCES
1 Anderson, J. B. and Mason, William T. Jr.
A Comparison of Benthic Macro-
invertebrates collected by Dredge and
Basket Sampler. Jour. Water Poll.
Cont. Fed. 40(2) 252-259.
2 Arthur, John W. and Horning, W. B , II.
The Use of Artificial Substrates in
Pollution Surveys. Amer Midi. Nat.
82(1) 83-89.
3 Besch, W., Hoffman, W. , and Ellenberger,
W. Das Macrobenthos auf
Polyatchylensubstraten in Fliessgs-
wasseren. Annals de Limnologic.
3(2) 331-367 1967
4 Burbanck, W.D. and Spoon, D.M The
Use of Sessile Ciliates Collected in
Plastic Petri Dishes for Rapid
Assessment of Water Pollution.
J. Protozool. 14(4) 739-744. 1967.
5 Cooke, William B. Colonization of
Artificial Bare Areas by Microorganisms.
Bot Rev. 22(9) 613-638. Nov. 1956.
6 Fox, Alfred C. Personal Communication.
1969.
7 Hester, F.E. and Dendy, J.S.
A Multiple-Plate Sampler for Aquatic
Macroinverteb rates. Trans.Am.
Fish. Soc. 91(4) 420-421. April 1962.
8 Hilsenhoff, William L. An Artificial
Substrate Device for Sampling Benthic
Stream Invertebrates Limnology and
Oceanography 14(3) 465-471 1969.
9 Mason, W.T., Jr., Anderson, J. B., and
Morrison, G.E. A Limestone-Filled,
Artificial Substrate Sampler Float Unit
for Collecting Macroinvertebrates in
Large Streams. Prog. Fish-Cult
29 74. 1967.
10 Ray, D. L Marine Boring and Fouling
Organisms University of Washington
Press, Seattle. pp 1-536. 1959.
11 Sickel, James B. A Survey of the
Mussel Populations (Umonidae) and
Protozoa of the Altamaha River with
Reference to their Use in Monitoring
Environmental Changes MS Thesis.
Emory University 133 pp. 1969
12 Sladeckova, A. Limnological Investigation
Methods for the Periphyton ("Aufwuchs")
Community. Bot. Rev. 28(2) 286-350.
1962.
13 Spoon, D.M. and Burbanck, WD. A
New Method for Collecting Sessile
Ciliates in Plastic Petri Dishes with
Tight Fitting Lids. J. Protozool
14(4) 735-739. 1967.
-------
Artificial Substrates
14 Visscher, J. Paul. Nature and Extent
of Fouling of Ships Bottom. Dept.
Comm. Bur. Com. Fish. Doc.
No. 1031. pp 193-252. 1928.
15 Weber, C.E. and Rauschke, R. L.
Use of a Floating Periphyton Sampler
for Water Pollution Surveillance.
Water Poll. Sur. Sept. Applications
and Develop. Report No. 20.
FWPCA -USDI, Cincinnati, Ohio.
September 1966.
16 Wene, George and Wickliff, E. L.
Modification of a Stream Bottom and
its Effect on the Insect Fauna.
Canadian Entomologist. Bull. 149,
5 pp. 1940.
17 Ziebell, Charles D., McConnell, W. J.,
and Baldwin, Howard A. A Some
Recovery Device for Submerged
Equipment. Limnol. and Ocean.
13(1):198-200. 1968.
This outline was prepared by H. W. Jackson,
Chief Biologist and R. M. Sinclair, Aquatic
Biologist, National Training Center, Water
Programs Operations, EPA, Cincinnati,
OH 45268.
-------
ATTACHED GROWTHS
(Periphyton or Aufwuchs)
I The community of attached microscopic
plants and animals is frequently investigated
during water quality studies. The attached
growth community (periphyton) and suspended
growth community (plankton) are the principal
primary producers in waterways--they con-
vert nutrients to organic living materials and
store light originating energy through the
processes of photosynthesis. In extensive
deep waters, plankton is probably the pre-
dominant primary producer. In shallow lakes,
ponds, and rivers, periphyton is the predominant
primary producer. During the past two
decades, investigators of microscopic
01 ganisms have increasingly placed emphasis
on periphytic growths because of inherent
advantages over the plankton when interpreting
data from surveys on flowing waters
A Blum (1956) . . .workers are generally
agreed that no distinctive association of
phytoplankton is found in streams, although
there is some evidence of this for individual
zooplankters (animals) and for a few
individual algae and bacteria. Plankton
organisms are often introduced into the
current from impoundments, backwater
areas or stagnant arms of the stream....
Rivers whose plankton is not dominated by
species from upstream lakes or ponds are
likely to exhibit a majority of forms which
have been derived from the stream bottom
directly and which are thus merely
facultative or opportunistic plankters. "
B "The transitory nature of stream plankton
makes it nearly impossible to ascertain at
which point upstream agents producing
changes in the algal population were
introduced, and whether the changes
occurred at the sampling site or at some
unknown point upstream. In contrast,
bottom algae (periphyton) are true com-
ponents of the stream biota. Their
sessile-attached mode of life subjects
them to the quality of water continuously
flowing over them. By observing the
longitudinal distribution of bottom algae
within a stream, the sources of the agents
producing the change can be traced
(back-tracked)" (Keup, 1966).
II TERMINOLOGY
A Two terms are equally valid and commonly
in use to describe the attached community
of organisms. Periphyton literally means
"around plants" such as the growths over-
growing pond-weeds, through usage this
term means the attached film of growths
that rely on substrates as a "place-to-
grow" within a waterway. The components
of this growth assemblage consists of
plants, animals, bacteria, etc. Aufwuchs
is an equally acceptable term [probably
originally proposed by Seligo (1905)] .
Aufwuchs is a German noun without
equivalent english translation, it is
essentially a collective term equivalent
to the above American (Latin root) term -
Periphyton. (For convenience, only,
PERIPHYTON, with its liberal modern
meaning will be used in this outline.)
B Other terms, some rarely encountered in
the literature, that are essentially
synonymous with periphyton or describe
important and dominant components of the
periphytic community are Nereiden,
Bewuchs, Laison, Belag, Besatz, attached,
sessile, sessile-attached, sedentary,
seeded-on, attached materials, slimes,
slime-growths, and coatings.
The academic community occasionally
employs terminology based on the nature
of the substrates the periphyton grows on
(Table 1).
TABLE 1
Periphyton Terminology Based
on Substrate Occupied
Substrate Adjective
various epiholitic, nereiditic, sessile
plants epiphytic
animals epizooic
wood epidendritic, epixylomc
rock epilithic
[After Srameck-Husek (1946) and via Sladeckova
< 1962)J Most above listed latin-root adjectives
are derivatives of nouns such as epihola,
epiphyton, spizoa, etc.
BI. MIC. enu. 19b. 5. 71
-------
Attached Growths (Periphyton or Aufwuchs)
III Periphyton, as with all other components V
of the environment, can be sampled quali-
tatively (what is present) and quantitatively A
(how much or many are present).
A Qualitative sampling can be performed by
many methods and may extend from direct
examination of the growths attached to a
substrate to unique "cuttings" or scrapings.
It may also be a portion of quantitative
sampling. B
B Quantitative sampling is difficult because
it is nearly impossible to remove the
entire community from a standardized or
unit area of substrate.
1 Areas scraped cannot be determined
precisely enough when the areas are
amorphous plants, rocks or logs that C
serve as the principal periphyton
substrates.
2 Collection of the entire community within
a standard area usually destroys individual
specimens thereby making identification
difficult (careful scraping can provide
sufficient intact individuals of the species
present to make qualitative determinations), VI
or the process of collection adds sufficient
foreign materials (1. e. detritus, sub-
strate, etc. ) so that some commonly A
employed quantitative procedures are
not applicable.
B
IV Artificial substrates are a technique
designed to overcome the problems of direct
sampling. They serve their purpose, but
cannot be used without discretion. They are
objects standardized as to surface area,
texture, position, etc. that are placed in the
waterway for pre-selected time periods during
which periphytic growths accumulate. They C
are usually made of inert materials, glass
being most common with plastics second in
frequency. Over fifty various devices and
methods of support or suspension of the
substrates have been devised (Sladeckova,
1962) (Weber, 1966) (Thomas, 1968).
ARTIFICIAL SUBSTRATE PLACEMENT
Position or Orientation
1 Horizontal - Includes effects of settled
materials.
2 Vertical - Eliminates many effects of
settled materials.
Depth (light) - A substrate placed in lighted
waters may not reflect conditions in a
waterway if much of the natural substrate
(bottom) does not receive light or receives
light at reduced intensity. (Both excessive
light and a shortage of light can inhibit
growths and influence the kinds of organisms
present. )
Current is Important
1 It can prevent the settling of smothering
materials.
2 It flushes metabolic wastes away and
introduces nutrients to the colony.
THE LENGTH OF TIME THE SUBSTRATE
IS EXPOSED IS IMPORTANT.
The growths need time to colonize and
develop on the recently introduced
substrate.
Established growths may intermittently
break-away from the substrate because
of current or weight induced stresses, or
"over-growth" may "choke" the attachment
layers (nutrient, light, etc. restrictions)
which then weaken or die allowing release
of the mass.
A minimum of about ten days is required
to produce sufficient growths on an
artificial substrate, exposures exceeding
a longer time than 4-6 weeks may produce
"erratic results" because of sloughing or
the accumulation of senile growths in
situations where the substrate is
artificially protected from predation and
other environmental stresses.
-------
Attached Growths (Periphyton or Aufwuchs)
VII Determining the variety of growths present
is presently only practical with microscopic
examination. (A few micro-chemical pro-
cedures for differentiation show promise--
but, are only in the early stages of development. )
VIH DETERMINING THE QUANTITY OF
GROWTH(S)
Nutrient analyses serve as indices of
the biomass by measuring the quantity
of nutrient incorporated.
a Carbon
1) Total organic carbon
2) Carbon equivalents (COD)
A Direct enumeration of the growths while
attached to the substrate can be used, but
is restricted to the larger organisms
because (1) the problem of maintaining
material in an acceptable condition under
the short working distances of the objective
lenses on compound microscopes, and
(2) transmitted light is not adequate
because of either opaque substrates and/or
the density of the colonial growths.
B Most frequently, periphyton is scraped
from the substrate and then processed
according to several available procedures,
the selection being based on the need, and
use of the data.
1 Aliquots of the sample may be counted
using methods frequently employed in
plankton analysis.
a Number of organisms
b Standardized units
c Volumetric units
d Others
2 Gravimetric
a Total dry weight of scrapings
b Ash-free dry weight (eliminates
inorganic sediment)
c A comparison of total and ash-free
dry weights
3 Volumetric, involving centrifugation of
the scrapings to determine a packed
biomass volume.
b Organic nitrogen
c Phosphorus - Has limitations
because cells can store excess
above immediate needs.
d Other
5 Chlorophyll and other bio-pigment
extractions.
6 Carbon-14 uptake
7 Oxygen production, or respiratory
oxygen demand
IX EXPRESSION OF RESULTS
A Qualitative
1 Forms found
2 Ratios of number per group found
3 Frequency distribution of varieties
found
4 Autotrophic index (Weber)
5 Pigment diversity index (Odum)
B Quantitative
1 Area! basis--quantity per square
inch, foot, centimeter, or meter.
For example
a 16 mgs/sq. inch
b 16, 000 cells/sq. inch
2 Rate basis. For example
a 2 mgs/day, of biomass accumulation
b 1 mg Og/mg of growth/hour
-------
Attached Growths (Penphyton or Aufwuchs)
REFERENCES
1 Blum, J.L. The Ecology of River Algae.
Botanical Review. 22 5-291. 1956.
2 Dumont, H. J. A Quantitative Method for
the Study of Penphyton. Limnol.
Oceanogr. 14(2):584~595.
3 Keup, L.E. Stream Biology for Assessing
Sewage Treatment Plant Efficiency.
Water and Sewage Works. 113-11~411.
1966.
4 Seligo, A. Uber den Ursprung der
Fischnahrung. Mitt. d. Westpr.
Fisch. -V. 17:4-52. 1905.
5 Sladeckova, A. Limnological Investigation
Methods for the Penphyton Community.
Botanical Review. 28-2.286. 1962.
6 Srameck-Husek. (On the Uniform
Classification of Animal and Plant
Communities in our Waters).
Sbornik MAP 20 3 213. Orig. in
Czech. 1946.
7 Thomas, N.A. Method for Slide
Attachment in Penphyton Studies.
Manuscript. 1968.
8 Weber, C.I. Methods of Collection and
Analysis of Plankton and Penphyton
Samples in the Water Pollution
Surveillance System. Water Pollution
Surveillance System Applications and
Development Report No. 19, FWPCA,
Cincinnati. 19+pp. (multilith). 1966.
9 Weber, C.I. Annual Bibliography
Midwest Benthological Society.
Penphyton. 1014 Broadway,
Cincinnati, OH 45202.
10 Hynes, H.B.N. The Ecology of Running
Waters. Univ. Toronto Press.
555 pp. 1970.
This outline was prepared by Lowell E. Keup,
Chief, Technical Studies Branch, Division of
Technical Support, EPA, Washington, DC 20242.
-------
APPLICATION OF BIOLOGICAL DATA
I ECOLOGICAL DATA HAS TRADITIONALLY
BEEN DIVIDED INTO TWO GENERAL
CLASSES'
A Qualitative - dealing with the taxonomic
composition of communities
B Quantitative - dealing with the population
density or rates of processes occurring
in the communities
Each kind of data has been useful in its own
way.
II QUALITATIVE DATA
A Certain species have been identified as
1 Clean water (sensitive) or oligotrophic
2 Facultative, or tolerant
3 Preferring polluted regions
(see Fjerdinstad 1964, 1965, Gaufin
& Tarzwell 1956, Palmer 1963, 1969,
Rawson 1956, Telling 1955)
B Using our knowledge about ecological
requirements the biologist may compare
the species present
1 At different stations in the same river
(Gaufm 1958) or lake (Holland 1968)
2 In different rivers or Jakes (Robertson
and Powers 1967)
or changes m the species in a river or/lake
over a period of several years. (Carr
& Hiltunen 1965, Edmondson & Anderson
1956, Fruh, Stewart, Lee & Rohlich 1966,
Hasler 1947).
C Until comparatively recent times taxonomic
data were not subject to statistical treat-
ment
III QUANTITATIVE DATA: Typical
Parameters of this type include
2
A Counts - algae/ml, benthos/m ,
fish/net/day
3
B Volume - mm algae /liter
C Weight - dry wgt, ash-free wgt
D Chemical content - chlorophyll,
carbohydrate, ATP, DNA, etc
E Calories (or caloric equivalents)
F Processes - productivity, respiration
IV Historically, the chief use of statistics
in treating biological data has been in the
collection and analysis of samples for these
parameters. Recently, many methods have
been devised to convert taxonomic data into
numerical form to permit-
A Better communication between the
biologists and other scientific disciplines
B Statistical treatment of taxonomic data
C In the field of pollution biology these
methods include-
1 Numerical ratings of organisms on the
basis of their pollution tolerance
(saprobic valency: Zelinka & Sladecek
1964)
(pollution index: Palmer 1969)
2 Use of quotients or ratios of species in
different taxonomic groups (Nygaard
1949)
BI.EN.3a. 12.70
-------
Application of Biological Data
3 Simple indices of community diversity
j Information theory
Organisms are placed in taxonomic
groups which behave similarly under
the same ecological conditions. The
number of species in these groups
found at "healthy" stations is com-
pared to that found at "experimental"
stations. (Patrick 1950)
The basic equation used for
information theory applications was
developed by Margalef (1957).
_1 . N'
N 2 N ' N, 1. . . N '
b A truncated log normal curve is
plotted on the basis of the number
of individuals per diatom species
(Patrick, Hohn, & Wallace 1954)
where I - information/individual,
N , N,. . .N are the number of
individuals in species a, b, ...
s, and N is their sum.
c Sequential comparison index.
(Cairns, Albough, Busey & Chanay
1968). In this technique, similar
organisms encountered sequentially
are grouped into "runs".
total organisms examined
d Ratio of carotenoids to chlorophyll
in phytoplankton populations-
OD430/OD665(Margalef 1968)
OD435^OD67Q(Tanaka, et al 1961)
e The number of diatom species present
at a station is considered indicative
of water quality or pollution level.
(Williams 1964)
number of species (S)
number of individuals (N)
number of species (S)
® square root of number of individuals (s/ N)
h ^(Menhinick 1964)
logg N
E n. (n - 1) (Simpson 1949)
1 H = 1 1
N (N - 1)
where n = number of individuals
belonging to the l-th species,
and
N = total number of individuals
This equation has also been used
with
1) The fatty acid content of algae
(Mclntire, Tinsley, and Lowry
1969)
2) Algal productivity (Dickman 1968)
3) Benthic biomass (Wilhm 1968)
REFERENCES
1 Cairns, J , Jr., Albough, D.W.,
Busey, F, and Chaney, M.D.
The sequential comparison index -
a simplified method for non-biologists
to estimate relative differences in
biological diversity in stream pollution
studies. J. Water Poll. Contr Fed
40(9).1607-1613 1968.
2 Carr, J.F and Hiltunen, J.K. Changes
m the bottom fauna of Western Lake
Erie from 1930 to 1961. Limnol.
Oceanogr. 10(4):551-569. 1965
3 Dickman, M. Some indices of diversity.
Ecology 49(6).1191-1193. 1968
-------
Application of Biological Data
4 Edmondson, W.T. and Anderson, G.C
Artificial Eutrophication of Lake
Washington. Limnol. Oceanogr.
1(1)47-53. 1956
5 Fjerdingstad, E. Pollution of Streams
estimated by benthal phytomicro-
organisms. I. A saprobic system
based on communities of organisms
and ecological factors Internat'l
Rev. Ges. Hydrobiol. 49(1) 63-131 1964
6 Fjerdingstad, E. Taxonomy and saprobic
valency of benthic phytomicro-
organisms. Hydrobiol. 50 (4) 475-604
1965
7 Fruh, E G , Stewart, K. M,, Lee, G, F
and Rohlich, G.A. Measurements of
eutrophication and trends. J Water
Poll Contr. Fed. 38<8)• 1237-1258
1966.
8 Gaufin, A.R. Effects of Pollution on a
midwestern stream. Ohio J. Sci.
58(4) 197-208 1958.
9 Gaufin, A.R and Tarzwell, C M. Aquatic
macroinvertebrate communities as
indicators of organic pollution in Lytle
Creek. Sew. Ind. Wastes. 28(7) 906-
924. 1956,
10 Hasler, A.D Eutrophication of lakes by
domestic drainage. Ecology 28(4) 383-
395. 1947
11 Holland, R.E Correlation of Melosira
species with trophic conditions in Lake
Michigan. Limnol. Oceanogr.
13(3) 555-557. 1968
12 Margalef, R. Information theory in
ecology. Gen. Syst 3 36-71 1957.
13 Margalef, R. Perspectives in ecological
theory. Univ. Chicago Press. 1968.
14 Mclntire, C.D., Tmsley, I J. and
Lowry, R. R Fatty acids in lotic
penphyton another measure of
community structure. J Phycol.
5 26-32. 1969
15 Menhinick, E. F. A comparison of some
species - individuals diversity indices
applied to samples of field insects.
Ecology 45-859. 1964,
16 Nygaard, G. Hydrobiological studies m
some ponds and lakes II. The
quotient hypothesis and some new or
little-known phytoplankton organisms
Klg Danske Vidensk. Selsk Biol
Skrifter 7 1-293. 1949.
17 Patten, B.C. Species diversity in net
plankton of Raritan Bay J. Mar.
Res. 20 57-75. 1962.
18 Palmer, C.M. The effect of pollution on
river algae. Ann. New York Acad.
Sci. 108-389-395. 1963
19 Palmer, C. M. A composite rating of
algae tolerating organic pollution.
J Phycol. 5(1) 78-82. 1969.
20 Patrick, R., Hohn, M.H. and Wallace,
J H. A new method for determining
the pattern of the diatom flora. Not.
Natl. Acad. Sci , No. 259
Philadelphia 1954
21 Rawson, D. S. Algal indicators of trophic
lake types. Limnol. Oceanogr.
1 18-25. 1956.
22 Robertson, S. and Powers, C.F
Comparison of the distribution of
organic matter in the five Great Lakes
m: J. C Ayers and D. C. Chandler,
eds. Studies on the environment and
eutrophication of Lake Michigan.
Spec. Rpt. No. 30, Great Lakes Res.
Div ,Inst. Sci. & Techn., Univ.
Michigan, Ann Arbor. 1967
23 Simpson, E.H. Measurement of diversity.
Nature (London) 163 688. 1949.
24 Tanaka, O.H , Irie, S Izuka, and Koga, F.
The fundamental investigation on the
biological productivity in the Northwest
of Kyushu. I. The investigation of
plankton. Rec. Oceanogr. W. Japan,
Spec. Rpt. No 5, 1-57 1961.
-------
Application of Biological Data
25 Telling, E. Some mesotrophic phyto-
plankton indicators. Proc. Intern
Assoc. Limnol. 12 212-215 1955.
26 Wilhm, J L. Comparison of some
diversity indices applied to populations
of benthic macroinvertebrates in a
stream receiving organic wastes. J
Water Poll. Contr Fed 39(10) 1673-1683
1967.
27 Wilhm, J L. Use of biomass units in
Shannon's formula, Ecology 49 153-156.
1968
28 Williams, L.G. Possible relationships
between diatom numbers and water
quality Ecology 45(4) 810-823. 1964
29 Zelmka, M. and Sladecek, V Hydro-
biology for water management.
State Publ. House for Technical
Literature, Prague 122 p 1964.
This outline was prepared by C I. Weber,
Chief, Biological Methods Section, Analytical
Quality Control Laboratory, NERC, EPA,
Cincinnati, OH 45268.
-------
PROCEDURES FOR FISH KILL INVESTIGATIONS
I INTRODUCTION
Fish kills in natural waters, though un-
fortunate, can in many instances indicate
poor water quality leading to investigations
which may improve water quality. Prompt
investigations should be organized and
conducted so that the resultant data implicates
the correct cause. Fish kills tend to be
highly controversial, usually involving the
general public as well as a number of
agencies Therefore, the investigator can
expect his findings to be disputed, quite
possibly in a court of law
The following procedures are presented as
a working guide for investigating and re-
porting fish kills as developed by the
personnel of The Lower Mississippi River
Comprehensive Project (FWPCA).
II TYPES AND EXTENT OF FISH KILLS
A Natural Mortalities - Those which are
caused through natural phenomena such
as, acute temperature change, storms,
ice and snow cover, decomposition of
natural materials, salinity change,
spawning mortalities, parasites, and
bacterial or viral epidemics.
B Man caused fish kills - Produced by
environmental changes through man's
activity, and may be attributed to
municipal wastes, industrial wastes,
agricultural activities and water control III
activities.
C One dead fish in a stream may be called A
a fish kill, however, in a practical
sense some minimal range in number of
dead fish observed plus additional
qualifications should be used in reporting
and classifying fish kill investigations.
The following definitions should be used
as guidelines in reporting fish kill
investigations. These qualifications
are based on a stream approximating
200 feet in width and 6 feet in depth.
For other size streams, adjustments
should be made.
1 Minor fish kill considered here as
NO fish kill and reported so-
1-100 dead or dying fish confined
to a small area or stream stretch.
Providing this is not a reoccurring
or periodic situation. For
example, near a waste outfall in
which stream dilution plays its
part and nullifies the effect of the
deleterious material. If this is a
reoccurring situation, it could be
of major significance and, there-
fore, investigated.
2 Moderate fish kill- 100 - 1000
dead or dying fish observed. In a
stream where dilution has had the
chance to play its role involving
a mile or so of stream , a number
of species are affected, and
apparently normal fish can be
collected immediately downstream
from the observed kill area.
3 Heavy fish kill 10, 000 fish or
more observed dead or dying.
In a stream where dilution has
had the chance to play its part,
but ten miles or more of the
stream are involved, many
species of fish are affected and
dying fish may still be observed
downstream.
PREPARATION FOR FIELD
INVESTIGATION
Secure maps of area to be investigated.
1 U.S. Geological Survey maps
a 1/250, 000 scale for general
location
b 1/24.000 for accurately
defining the kill area in the
field
2 Navigation maps (appropriate
agency)
BI. FI. 13d. 9. 72
-------
Procedures for Fish Kill Investigations
3 Other sources
4 From the data received from the
reporting agency, locate the kill
area on the map.
a Determine best access points.
b Locate possible known industries,
municipalities, or other
potential sources of pollution.
c Estimate the possible area to
be traveled or inspected on
1) water
2) land
3) both
Secure sampling equipment and deter-
mine size of investigation team needed.
of "Standard Methods" for specific
physical and chemical equipment
required for collecting, analyzing,
or preserving samples possibly
containing the suspected causative
agent.
Form an investigating party
a If only one man is available
to make the investigation,
preference for choosing the
man should be in this order
1) Specialized professional
personnel, such as,
engineer, chemist, or
biologist who is ex-
perienced in investigating
fish kills and who is
capable of adequately
reporting the technical
aspects of the investigation.
1 Standard equipment to be taken on
all investigations (a standard
checklist with space for special
equipment will often save
embarrassment in the field.)
a Thermometer
b Dissolved oxygen sampler
c D.O. bottles
d Winkler D.O. test kit
e Conductivity test meter
f pH test meter or chemical kit
g Sample bottles
h Pencils and note paper
i Current edition of "Standard
Methods for the Examination
of Water and Wastewater "
2 If preliminary information is
available on the possible cause of
the kill, consult the latest edition
2) A non-specialized pro-
fessional engineer,
chemist, or biologist who
has little or no experience
in fish kill investigations,
but who is capable of
adequately reporting the
technical aspects of the
investigation.
3) A technician who has
considerable field
experience in pollution
and fish kill investigations
and who is capable of
reporting some of the
technical aspects of the
investigation.
4) An office technician or
other personnel who has
had limited field work in
pollution investigations.
b If two or more men are needed
for the investigation, the party
should include at least one
person under category (1)
above. Preferably, the team
should include:
-------
Procedures for Fish Kill Investigations
1) A biologist to make a
survey of the biological
changes.
2) An engineer to make an
evaluation of the physical
condition of the fish kill
area and to make an
investigation of an industry
or a municipal wastewater
treatment plant if needed.
c If a fish kill is observed in its
initial state m the field by any
one of the people listed under
the classification in Section
B.3.a., the project office
should be informed immediately
(after working hours the project
director or deputy director
should be informed) so that an
adequately equipped, specialized
investigating party can be
formed if needed.
C Contact personnel of the laboratory or
laboratories which will participate in
analyzing samples. If possible estimate
the following and record on sample form
No. 1.
1 The number and size of samples to
be submitted
2 The probable number and types of
analyses required
3 The dates the samples will be
received by the laboratory
4 Method of shipment to the laboratory
5 To whom the laboratory results are
to be reported
6 The date the results are needed
IV MAKING THE FIELD INVESTIGATION
A Contact the local lay person or official
who first observed the kill and reported
it.
1 Obtain any additional information
which might be helpful which was
not reported previously.
2 If possible, retain the reporting
party as a guide or invite him to
accompany the investigating team.
B Make a reconnaissance of the kill area.
1 Make a decision as to the extent
of the kill and if a legitimate kill
really has occurred.
2 If a legitimate kill exists take steps
to trace or determine the cause.
a Always perform the following
physical or chemical tests,
during the initial steps of the
investigation
1) Temperature
2) pH
3) Dissolved oxygen
4) Specific conductance
While none of these factors
may be directly involved in
the kill these tests are per-
formed simply and rapidly in
the field and can be used as a
baseline or starting point for
isolating the cause (s) of the
kill.
b Record other physical
observations such as.
1) Appearance of water, i.e.,
turbidity, high algal
blooms, oily, unusual
appearance, etc.
2) Stream flow pattern, i.e.,
high or low flow, stagnant
or rapidly moving water,
tide moving in or out, etc.
If possible obtain reading
from stream gage if one
is near kill area.
-------
Procedures for Fish Kill Investigations
3) Weather conditions pre-
vailing at the time of the
investigation and information
on weather immediately
prior to the kill
3 Make a rough sketch or define the
kill area on a map so that sampling
points, sewer outfalls, etc. can be
accurately located on a drawing to
be included in a final report.
4 Take close-up and distance
photographs of:
a Dead fish in the stream in the
polluted area.
b The stream above the polluted
area.
c Wastewater discharges.
Photographs will often show a
marked delineation between the
wastewater discharge and the
natural flow of water. Pictures
taken at a relatively high elevation,
(a bridge as opposed to a boat or
from a low river bank) will show
more and be more effective.
Color photographs are also more
effective in showing physical con-
ditions of a stream in comparison
to black and white prints.
C Sampling Procedures - The extent and
method of sampling will depend upon
location and upon the suspected cause of
the kill.
1 Stream and wastewater sampling.
a Sample the following points when
the pollutional discharge is
coming from a well defined
outfall.
1) The effluent discharge
outfall
2) The stream at the closest
point above the outfall which
is not influenced by the
waste discharge
3) The stream immediately
below the outfall
4) Other points downstream
needed to trace the extent
of the pollution
b The sampling should be ex-
tensive enough that when all
the data is compiled no question
will exist as to the source of
the pollution which killed the
fish.
c The number of samples to be
collected at a given cross
section will depend principally
on the size of the stream.
1) Streams less than 200 feet
wide, not in an industrial
area usually can be
adequately sampled at one
point in a section (Figure 1).
2) Streams 200 feet or wider
generally should be
sampled two or more
places in a section
immediately above and
below the pollutional
discharge, where the
pollutional waste has
adequately mixed with the
stream flow one sample
may suffice.
3) A number of samples m a
cross section may be
required on any size of
stream to show that the
suspected pollutional
discharge is coming from
a source located in an
industrial or municipal
complex (Figure 2).
4) Extensive cross sectional
sampling on rivers
greater than 200 feet wide
will be required for kills
involving suspected
agricultural or other types
of mass runoff.
-------
Procedures for Fish Kill Investigations
Suspected source of
pollution.
Areo of dead fish and/or
obvious pollution discharge.
Direction of - »
—— > tf)
^0)
8 s
OJ °
Bridge
Figure I — Minimum Water Sampling Point On Stream 200 Feet Or Less
Wide Involving An Isolated Discharge.
Discharge sources relatively close
to suspected source of pollution
s&X-FD
SIM
A
Direction of
flow
uspecled source
of pollution
Bridge
E
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n ci
s-
< ?
4-
11 g • i 119
|
III
Figure 2 — Minimum Water Sampling Points On A Stream Running
Through An Industrial Or Municipal Complex.
PLATE I - RELATIONSHIP OF FISH KILLS TO SOURCE OF TOXICITY
-------
Procedures for Fish Kill Investigations
5) Sample depths - On streams
5 feet in depth or less, one
mid-depth sample per
sampling locations. For
streams of greater depths,
appropriate sampling
judgment should be used,
since stratification may be
present.
Explanation of Plate I
1) Collection point 1, Figure
1 and points 3 and 4,
Figure 2 should be collected
as near to the point of
pollutional discharge as
possible. These points
will vary according to
stream flow conditions; the
pollutional discharges into
a slow'sluggish stream
usually will have a cone of
influence upstream of the
outfall, whereas, a swift
flowing stream usually will
not.
2} Collectmg an upstream
control sample from a
bridge within sight of the
pollutional discharge
would probably be satis-
factory in Figure 1 but
definitely not m Figure 2.
3) Figures 1 and 2 are given
for illustrative purposes
only and should be used
only as a guide for sampling.
Thought must be given to
each individual situation
to insure adequate, proper
sampling. While too many
samples are better than
too few, effort should be
made not to unduly over-
load the laboratory with
samples collected as a
result of poor sampling
procedures.
2 Biological sampling
a In every investigation of fish
or wildlife kills the paramount
item should be the immediate
collection of the dying or only
recently dead organism.
This may be done by anyone,
sampling and preservation is
as follows-
1) Collect 20 plus drops of
blood in a solvent rinsed
vial, seal same with
aluminum foil, cap and
freeze.
2) Place bled carcass, or
entire carcass if beyond
bleeding stage, in plastic
bag and freeze. In case
no method of freezing is
available, icing for a
short period prior to
freezing may be acceptable.
Labeling of both blood and
carcass is important.
3) Controls-live specimens
of the affected organisms
should be obtained from
an area within the same
body of water which had
not been influenced by the
causative agent. Once
obtained these specimens
should be handled in a
like manner.
b The number of individuals
involved and the species
affected should be enumerated
in some manner. At most
these will be estimates.
Depending on the given situation
such as area or distance
involved and personnel available
enumeration of fish kills may
be approached in one of the
following ways.
-------
Procedures for Fish Kill Investigations
1) For large rivers, establish
observers at a station or
stations (e. g., bridges}
and count the dead and/or
dying fish for a specified
period of time, then pro-
ject same to total time
involved.
2) For large rivers and Jakes,
traverse a measured
distance of shoreline,
count the number and
kinds of dead or dying fish.
Project same relative to
total distance of kill.
Other aspects of the biota
which should be considered
are the aquatic plants. In lakes
and ponds floating and rooted
plants should be enumerated
and identified. The collection
of plankton (rivers and lakes)
should be taken in order to
determine the degree of bloom,
which in itself may cause fish
kills because of diurnal DO
levels.
Both aquatic plants and macro-
invertebrates may be preserved
in a 5% formalin solution.
3) For lakes and large ponds, 4 Bioassay
count the number and species
within measured areas, and Static bioassay techniques as out-
then project to total area lined in Standard Methods may be
involved. effectively used to determine acute
toxicity of wastes as well as
4) For smaller streams one receiving waters,
may walk the entire
stretch involved and count a In situ using live boxes
observed number of dead
individuals by species. b Mobile bioassay laboratory
3 Biological sampling Macro-
Invertebrates
a Sampling of benthic organisms
after the more urgent aspects
of the kill investigation has been
completed can prove to be
rewarding relative to extent and
cause of kill. Since this general
form of aquatic life is somewhat
sedentary by nature any release
of deleterious materials to their
environment will take its toll.
Thus by making a series of
collections up and downstream,
the affected stretch of stream
may be delineated when the
benthic populations are compared
to those from the control area.
Also the causative agent may
be realized when the specifics
of the benthic population present
are analyzed.
c Samples returned to Central
Lab for toxicity tests
V DETAILED EXAMINATION OF SOURCE
OF POLLUTION
A Seven general categories under which
causes of kills can be grouped are:
1 Industrial waste discharges
2 Waste discharges from municipal
sewerage systems
3 Water treatment plant discharges
4 Agriculture and related activities
5 Temporary activities
6 Accidental spills of oil and other
hazardous substances
-------
Procedures for Fish Kill Investigations
7 Natural causes
B Industrial Waste Discharges
1 Upon locating the outfall source,
collect a sample immediately if
possible at the point where the
wastes leave the company property.
2 Make an in-plant inspection if
possible.
a Contact the plant manager or
person in charge
b Request a brief tour of the
facilities
c Obtain general information
concerning the products
manufactured, raw materials,
manufacturing process,
quantities, sources, and
characteristics of wastes, and
waste treatment facilities if any
Possibly the company may be
able to supply a flow diagram
or brochure of the plant
operations.
d Request specific information
concerning the plant operation
immediately prior to the start
of the kill
C Waste discharges from a municipal or
domestic type sewerage system
1 Discharges from this source may
be domestic sewage and industrial
wastes combined with domestic
sewage. These wastes may be
subjected to treatment of a municipal
treatment plant or may be dis-
charged directly, untreated to a
stream
2 Generally, the municipality or
owner of the sewerage system is
held responsible for any discharge
in such a system, consequently,
after collecting samples, the owner
or a representative of the owner of
the sewerage system should be
contacted. This may be a sewage
treatment plant operator, city
engineer, public works supervisor,
a subdivision developer, etc
a Obtain information about the
operation of the system
b If the cause of the kill was the
result of an industrial waste
discharge to a municipal
sewer and thence to a stream,
information should be obtained
from a municipal official about
the industry and the problem.
An inspection of the industrial
plant may be desirable.
Generally, this should be done
only in cooperation with a
municipal official
D Agriculture and Related Activities
1 Pollution capable of causing fish
kills may result from such
agricultural operations as crop
dusting and spraying fertilizer
applications, and manure or other
organic material discharges to a
stream
2 Generally, kills related to these
factors will be associated with
high rains and runoff.
3 The source or type of pollution
may be difficult or impossible to
locate exactly. It may involve a
large area. Talking to local
residents may help pinpoint the
specific problem area Runoff
from fields, drainage ditches, and
small streams leading to the kill
area are possible sampling places
which may be used to trace the
cause
E Temporary Activities
1 Causes of kills may result from
such temporary or intermittent
activities as mosquuo spraying,
construction activities involving
chemicals, oils, or other toxic
-------
Procedures for Fish Kill Investigations
substances, and weed spraying
with herbicide containing materials
toxic to fish such as arsenic.
2 As with agricultural activities,
tracing the cause of these kills is
difficult and may require extensive
sampling.
3 Accidental spills from ruptured
tank cars, pipelines, etc., and
dike collapse of industrial ponds
are frequently sources of fish kills.
F Possible Natural Causes of Fish Kills
1 Types of natural causes
a Oxygen depletion due to ice and
snow cover on surface waters
b Oxygen depletion at night because
of plant respiration or at any-
time during the day because of
natural occurring organics in
the water
c Abrupt temperature changes
d Epidemic and endemic diseases,
parasites, and other natural
occurring biological causes
e Lake water inversion during
vernal or autumnal turnover
which results in toxic material
or oxygen-free water being
brought to the surface
f Interval seiche movement in
which a toxic or low DO
hypolimnion flows up into a bay
or bayou for a limited period
of time, and later returns to
normal level
2 Fish kills in rivers below high dams
immediately following the opening
of a gate permitting hypolimnionic
water to flow down the stream
(as in TVA region)
VI CASE HISTORY
A The Lower Mississippi Endrin kill is an
excellent example of the investigation of
a major fish kill Bartsch and Ingram
give the following summary (See Table 1)
TABLE 1
ELEMENTS OF INVESTIGATIONS
I Examination of usual environmental
factors
~ Elimination of parasites, bacterial or
viral diseases and botulism as causes of
mortalities *
in Considerations of toxic substances
Examination and prognostication of
symptoms of dying fish Autopsy,
including
Haematocrits and white cell counts
Kidney tissue study
Brain tissue assay for organic
phosphorus insecticide
Tissue analysis for 19 potentially
toxic metals
Gas chromatographic analysis of
tissues, including blood, for
chlorinated hydrocarbon insecticides
IV Explorations for toxic substances
Bloassay with Mississippi River
water
Bioassay with extracts from river
bottom mud
Bloassay with tissue extracts from
fish dying in river water and
bottom mud extracts
Bioassay with endrin to compare
symptoms and tissue extract
analyses with those of dying fish in
all bioassays
V Intensive chemical analysis for
pesticides in the natural environment,
experimental environment, river fish,
and experimental animals
VI Surveillance of surface waters for
geographic range and intensity of
pesticide contamination
VII Correlation and interrelation of findings
* Th« invaailgawr should ba awaro of tha
(act ihit apparaotly healthy flah nuy b*
hirtoortaf pathosanlc bactarL* In (hair
bloodatraama
-------
Procedures for Fish Kill Investigations
The investigation was designed to con-
sider and eliminate potential fish kill
possibilities that were not involved and
come to a point focus on the real cause.
It was found that the massive kills were
not caused by disease, heavy metals,
organic phosphorus compounds, lack of
dissolved oxygen or unsuitable pH.
Blood of dying river fish was found to
have concentrations of endrin equal to or
greater than laboratory fish killed with
this pesticide, while living fish had
lesser concentrations. Symptoms of
both groups of dying fish were identical.
It was concluded from all data obtained
that these fish kills were caused by
endrin poisoning.
B Recent investigations in Tennessee have
shown that the leaking of small amounts
of very toxic chemicals from spent
pesticide-containing barrels used as
floats for piers and diving rafts in lakes
and reservoirs can produce extensive
fish kills. The particular compound
used to control slime growth in manu-
facturing processes, contained two
primary chemicals in solution
(phenylmercuric acetate and 2, 4,
6-trichlorophenol) The former com-
pound which breaks down to form
diphenylmercury was found to be more
toxic to aquatic life than the latter.
REFERENCES
1 American Public Health Association, Inc.
Standard Methods for the Examination
of Water and Wastewater Section 231
Bioassay, Examination of Polluted
Waters, Wastewaters, Effluents, Bottom
Sediments, and Sludges. Thirteenth
Edition. New York. 1971.
2 Bartsch, A.F. and Ingram, William N
Biological Analysis of Water Pollution
in North America International
Verein Limnol. 16-786-800. 1966
3 Bullock, G L. and Snieszko, S F
Bacteria m Blood and Kidney of
Apparently Healthy Hatchery Trout
Trans American Fisheries
Society 98(2) 268-27 1 1969
4 Burdick, G E Some Problems in the
Determination of the Cause of Fish
Kills. Biol Prob in Water
Pollution. USPHS Pub. No 999-
WP-25. dd. 289-292. 1965
5 Fish Kills Caused by Pollution in
1970. 11th Annual Report 21 p.
1972.
6 Mount, Donald I. and Putnicki, George J.
Summary Report of the 1963
Mississippi River Fish Kill
Investigation, 31st North American
Wildlife and Natural Res. Conf
11pp. 1966
7 Smith, L. L. Jr., et al. Procedures
for Investigation of Fish Kills
(A guide for field reconnaissance
and data collection) ORSANCO,
Cincinnati, OH. 24 pp. 1956
8 Tennessee Valley Authority Fish
Kill in Boone Reservoir. TVA
Water Quality Branch, Chattanooga
TN 61 pp. 1968.
9 Tennessee State Game and Fish
Commission. Field Manual for
Investigation of Pollution and Fish
Kills {USPHS WPD 3-0351-65
Grant) 71 pp. undated
10 Willoughby, L. G. Salmon Disease in
Windermere and the River Leven;
The Fungal Aspect. Salmon and
Trout Magazine. 186:124-130. 1969.
11 Muncy, Robert J. Observations on the
Factors Involved with Fish Mortality
as the Result of Dinoflagellate "Bloom"
in a Freshwater Lake Proc. 17th
Ann. Conf. Southeastern Assoc. of
Game & Fish Commissioners, pp.
218-222.
This outline was prepared by Jack Geckler,
Research Aquatic Biologist, Fish Toxicology
Activities, EPA, Newtown, OH 45244
-------
Project Personnel
Contacted
a. Name
b. Means of
Contact
c. Date & Time
1. Reporting source
a. Agency
(1) Address
(2) Phone (s)
b. Individual
(1) Address
(2) Phone
(3) Fish Kill Network yes no
c. Other Contacts
(1 > Address
<2) Phone
(3) Fish Kill Network yes no
2. Data furnished by reporting source
a. Location of Kill
b. Dates of Kill Dying last observed
c. Kinds of organisms
d. Approximate number killed
e. Cause of kill (if known)
f. Suspected causative sources
g. Measures taken
h. Other Agencies contacted
(1) Date and Time
3. Action requested
a. Field investigations
b. Laboratory analysis
4. Assistance to Project
a. Provided by
b Personnel
c. Equipment
d. Transportation facilities
-------
AN INITIATION INTO STATISTICS
I INTRODUCTION
Quantitative statistical analysis has been
publicized by some analyst as a cure all.
This exaggerated claim is adhered to by only
a few opportunists.
Quantitative analysis should be thought of as
a valuable tool which can be exploited by
close cooperation between the statistician
and the investigator in a joint attempt to find
better methods of controlling or understanding
the complex interaction of men, materials,
machines and the natural environmental
resources.
Modern statistics is still a relatively new
subject with along list of unsolved statistical
problems. Unless caution is observed the
inevitable result will be some unhappy
experiences with inadequately equipped
amateurs. In point of fact, most statistical
procedures are quite simple and intuitively
acceptable once they have been pointed out in
a particular context. Heuristic persuasion
can be effectively used to give insights into
the concept, development and foundations of
the particular tool used.
E IMPORTANCE OF PREPLANNING
Many investigators now believe that statistics
and particularly the statistical design of data
collection has potentially impressive leverage
effects on the amount of information to be
included and obtainable from any research
effort.
Statistical methods cannot reveal anything
that is not already implicit in the data.
However, every correct inference of
interest possible should be made. A
continual effort should be made to recover
all of the information but there never will
be a way to find more than is already there.
It would be better to do enough work on five
projects than an inadequate amount on a much
larger number. You don't have to plan in
order to fail, all you have to do is fail to
plan.
Many investigators still believe that
statisticians only enter at the end of the
work--that is, at the final examination of
the data. It is small comfort to the
investigator to be told after the event, that
the answers to his questions are inconclusive
and were bound to be so because an inadequate
amount of effort was allotted to the work.
It is well known that the end doesn't justify
the means but in data collection and analysis
the end dictates the means. The importance
of preplanning cannot be overemphasized.
If you fail to plan then you are planning to
fail.
Any investigation problem can be viewed as
a five step process. These are:
1 Problem definition
2 Data collection scheme
3 Actual data collection
4 Data analysis
After all, any finite body of data contains
only a limited amount of information. This
limit is set by the inherent nature of the data
themselves and cannot be increased by any
amount of ingenuity exercised by the data
analyst. However intelligent preplanning
can increase this limit.
5 Written report or recommendations
The most important step is the first one
because the second is based upon it. The
third follows from the second, etc. The
ST.51.10.72
-------
An Initiation Into Statistics
process is like a chain with five links. If the
first link is incorrect, the entire chain is.
Too often, too little time is spent on step one.
Many times the problem is vaguely or ill
defined. The best protection against these
faults is to write down black on white a state-
ment of the problem and have all connected
with the investigation to agree on the statement.
With this as an introduction we proceed to
describe the field of statistics briefly.
Ill ALTERNATIVE VIEWS OF STATISTICS
The field of statistic can be described from
many different angles. Each viewpoint results
in a different insight into this vast discipline.
In this introduction we consider the entire
field from three viewpoints. The first view is
very simple, the second introduces more
details and finally the last viewpoint is very
comprehensive with respect to subject matter
but is only one molecule deep.
IV FIRST VIEWPOINT
The first way of looking at statistics is
to partition the data source into the method
of acquisition as either experimental or
nonexperimental.
FIGURE 1
FOURFOLD MAP OF ENDS AND MEANS IN APPLIED STATISTICS
Source Or Means Of Data Acquisition
Nonexperimental
Experimental
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Tables and graphs
Averages central tendency
Dispersion measures
Frequency curves
Correlation
Survey sampling
Index numbers
Same
Assessment of probabilities
Theoretical means, variances,etc.
and corresponding confidence
intervals
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Specialized branches of
applied statistics
Demography
Econometrics
Genetics
Causal inference from
time series
Design of factorial experiments
Randomization methods
Significance tests
Regression analysis
Analysis of variance and
covariance
Probit analysis
-------
An Initiation Into Statistics
In an experimental investigation, control is
exercised over some or all of the influencing
variables, while in a nonexperimental
investigation nature is merely measured as
is without any control on any of the influencing
variables.
The purpose of the analysis is also viewed as
two-fold, that is, either enumerative or analytic,
The combination of double means and ends
leads to the four-fold table in Figure 1 This
is taken essentially from reference 1.
Historically, the field of statistics started
with the upper left quadrant about the turn
of the century. The research proceeded
clockwise until the lower left hand quadrant
was reached. That is causal inference from
observational data. This quadrant is the
most difficult and has been receiving the
most research attention lately.
In each quadrant some of the techniques are
indicated and the reader should be further
cautioned that the lines drawn are fluid and
are not rigid. A partial listing has been
given in each quadrant and no attempt has
been made to give a 100% enumeration.
V SECOND VIEWPOINT
The next viewpoint of statistics is in this
orientation the one taken by The International
Statistical Institute Figure 2 shows their
scheme for classification of abstracts Any
article in statistics can be classified in one
or more of the indicated titles. Some articles
are given a primary classification and others
in addition are given a secondary classification.
Let us give a brief introduction into the major
topics listed in Figure 2.
Upon inspection of the subtitles of the first
topic, mathematical methods, it is evident
that these subtitles are a list of the mathe-
matical techniques used by the professionals
to solve the difficult problems encountered
in the development of the theory and methods.
FIGURE 2
SCHEME FOR CLASSIFICATION
OF ABSTRACTS
0. MATHEMATICAL METHODS (White)
0.
General papers
1.
Solution of equations
2.
Methods of curve and surface fitting,
smoothing
3.
Interpolation and quadrature
4.
Special functions and transforms
5.
Functional relationships
6.
Determinantal and matrix analysis
7.
Game theory
8.
Programming techniques
9.
Group and field theory
10.
Graph theory and combinatorial
analysis
11.
Measure theory
12.
Optimisation
1. PROBABILITY (Pink)
0.
General papers
1.
Calculus of probabilities
2.
Expected values
3.
Combinatorial problems
4.
Geometric probability
5.
Limit theorems
6.
Stochastic convergence
7.
Stochastic approximation
8.
Decision theory and functions
9.
Transforms Fourier, Laplace, etc.
10.
Convolutions
2. FREQUENCY DISTRIBUTIONS (Green)
0. General papers
1. Descriptive properties
2. Transformations of vanatcs
3. Normal and lognormal
4. Binomial, multinomial and
hypergeometric
5. Poisson, exponential, negative
binomial, logarithmic and
contagious
6. Rectangular, extreme value and
Weibull
7. Pearson and "series expansion"
distributions
8. Truncated and mixed distributions
9. Multivariate distributions
10. Limit distributions
11. Approximations
12. Other distributions
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An Initiation Into Statistics
FIGURE 2
3. SAMPLING DISTRIBUTIONS (Light Blue)
0.
General papers
1.
t, z, F and x distributions
2.
Non-central distributions
3.
Studentisation
4.
Quadratic forms
5.
Correlation and regression
coefficients
6.
Location and scale statistics
7.
Shape and other descriptive
statistics
8.
Order statistics
9.
Multivariate problems
10.
Limit distributions
11.
Linear forms
4. ESTIMATION (Yellow)
0. General papers
1. Properties of estimators
2. Types of estimator Bayes,
maximum likelihoodj least
squares, etc.
3. Individual estimators point
4. Individual estimators: interval
5. Inequalities tolerance limits
and regions
6. Distribution-free methods
7. Sequential methods
8. Multivariate problems
9. Finite population procedures -
surveys
10. Simultaneous estimation
11. Distribution functions and densities
12. Decision theory
5. HYPOTHESIS TESTING (Purple)
0. General papers
1. Properties of test
2. Individual hypotheses
3. Two-sample problem
4. k-sample problem
5. Outliers
6. Distribution-free tests
7. Sequential tests
8. Multivariate problems
9. Types of test- likelihood ratio,
Bayes, minimax, etc.
10. Goodness-of-fit tests
11. Combining and comparing tests
12. Decision theory
6. RELATIONSHIPS (Grey)
0.
General papers
1.
Regression, linear hypothesis.
polynomials
2.
Correlation inc. canonical
correlation
3.
Factor methods and principal
components
4.
Discriminant analysis and clustei
analysis
5.
Ranking and scaling methods
6.
Systems of equations- structure
7.
Non-linear equations-logistic
8.
Transformed relationships-
quantal response
9.
Association and contingency
10.
Functional relationships
11.
Non-standard conditions
12.
Other multivariate methods
7. VARIANCE ANALYSIS (Biscuit)
0.
General papers
1.
Fixed effects model
2.
Variance components model
3.
Mixed and other models
4.
Non-orthogonal data and missing
values
5.
Non-standard conditions-failure
of assumptions
6.
Covariance analysis
7.
Multiple comparisons, multiple
decision procedures
8.
Ranked data
9.
Sequential methods inc. preliminary
tests
10.
Combining sets of results
11.
Precision of measurement
12.
Multivariate models
8. SAMPLING DESIGN (Orange)
0.
General papers
1.
Simple random, stratified,
multi-stage
2.
Sampling with unequal probability
3.
Multi-phase sampling, double
sampling
4.
Natural (human, animal and
biological) populations
5.
Non-sampling problems
6.
Censored, systematic and quota
sampling
7.
Nature and number of units, cost
and efficiency
8.
Acceptance inspection
9.
Process control
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An Initiation Into Statistics
FIGURE 2 (continued)
9. DESIGN OF EXPERIMENTS (Blue)
0. General papers
1. Block designs, designs for two-way
elimination of heterogeneity
2. Factorial arrangements
3. Response surfaces
4. Nature of unit, number of replications,
cost and efficiency
5. Paired comparisons and matching
problems
6. Preference tests
7. Repeated and sequential experiments
8. Weighing problems
9. Sensitivity problems
10. Systematic designs
11. Screening tests
12. Other designs, e.g., mixtures
10. STOCHASTIC PROCESSES AND TIME
SERIES (Red)
0. General papers
1. Properties of individual process
2. Estimation problems
3. Tests of hypotheses
4. Queuemg, storage, risk and congestion
theory
5. Information theory
6. Stationary processes and spectral
analysis
7. Auto and serial correlation
8. Multivariate processes
9. Biological population studies, genetic
models
10. Renewal theory
11. Markov processes
12. Branching processes
11. MISCELLANEOUS AND SPECIAL
TOPICS (Cream)
0. General statistical methodology
1. Statistical tables and charts
2. Probability graph papers
3. Nomograms and graphic methods
4. Machine methods, hand and punched
cards
5. Machine methods, electronic digital
6. Machine methods, other
7. Monte Carlo methods
8. Index numbers
9. History, biography and bibliography
10. Inventory
11. Life-testing and reliability
12. Teaching and training methods
These are the tools used to extend the
present boundaries of knowledge. They
are not statistics per se but are an indis-
pensable aid to the research statistician.
The next topic, probability, can be
described as a method for quantitizing
uncertainty where the limits of uncertainty
are a numerical value which does not lie
outside the range from zero to one. Most
people have a correct intuitive feel for
probability. If an event can happen (l. e.,
it will rain) say with probability 0. 9
(or 90%) then one knows that the event
could or could not occur. However, for
a large number of trials, on the average
nine times out of ten it will occur (or ram)
and one in ten it won't.
There is no universally acceptable abstract
calculus of probability. That is to say if
one adopted a set of axioms and developed
the theory of probability then the inevitable
result would be some controversy and many
heated arguments. Some fault would be
found in the development of the theory by
the purists. The purists are those who are
concerned with the philosophical and correct
logical foundations. They are necessary
and serve a useful purpose. They are not
boat rockers but keep it from sinking be-
cause they plug any holes that may be
present.
Most statisticians view probability from a
practical viewpoint and their attitude is
that it works and does a good job, so let
the purists thrash out the difficulties.
In the third topic, frequency distribution,
studies are concerned with characterizing
and identifying the distribution of a variable
or measurable characteristic. The most
famous of these distribution is the bell
shaped symmetric curve known as the
normal distribution. It is characterized
by two parameters, namely, the mean,
a measure of central tendency and the
standard deviation, a measure of spread
or variability.
-------
An Initiation Into Statistics
The list of distribution used in statistic
is greater than the subtitles listed. This
list includes only the most important ones
and a complete list of distributions
developed by researchers with their
properties would fill a book.
The usefulness of a frequency distribution
lies in the fact that from it one can calcu-
late the probability of occurrence of
different values or set of values for the
variable.
Very few people have the luxury when doing
research of measuring every unit in the
population. For this reason sampling
distribution per se are studied quite
vigorously. Constraints of time, money,
people and resources needed, versus
available, force researchers into taking
a sample or a subset of the entire
population. On the basis of a sample,
decision statements or estimates are
made about the entire population.
Different samples have different values or
a sample varies from one sample set to
another. Each sample as well as any value
calculated from it has its own sampling
distribution. These must be known before
quantitative statements can be made.
The three most important sampling distri-
butions are the t, F, and Chi-square.
They, as well as the normal, are tabulated
in almost every textbook on statistics.
In the fifth subject, estimation, the sample
values are used to infer information about
the entire population. For example, from
a sample, one can calculate the sample
mean and use this to make statements
about the parent population mean. The
calculated sample mean (X) is an estimate
of the population mean {the symbol used
is the Greek letter mu).
One legitimate question that can be asked
is "is this the best possible way?" Best
possible way is a fuzzy concept and must
be unfuzzied before the question can be
answered. That procedure produces many
interesting paths as evidenced by the list
of subtitles.
Hypothesis testing (or significance testing),
our next topic, is a technique that is too
often not really understood. It can be
understood by anyone since the concept
is really quite simple and is analogous
to a comparison procedure familiar to
everyone. A test of hypothesis is the
familiar method of comparing two things
(an unknown and a known) for identity or
nonidentity. After the comparison has
been completed then one of two conclusions
is made: 1) the two things compared are
not identical or 2) the things compared are
identical.
In the world of statistics, the unknown in
the comparison procedure is the sampled
population. The known used for the
comparison is a hypothetical population
which is also called the mathematical
model or the test assumptions.
The sampled population values are used
to evaluate a statistic which can be thought
of as a formula for combining the informa-
tion available in the sample. The observed
statistic value obtained from the sample
is compared with corresponding values
obtained from the mathematical model.
If the observed and expected statistic
values are in disagreement then the
sampled and hypothetical population
disagree or are not identical. If the
statistic values agree the populations
agree or are identical.
A simple example will make the ideas
clear. Assume you are sampling from
normal with an unknown mean and a
standard deviation equal to ten. We
wish to test if the unknown mean equal
five. The mathematical model or known
is normal, mean = 5 and standard
deviation = 10. If the conclusion of the
comparison procedure is agreement
between the sampled population and the
mathematical model. This means both
normals have equal means and standard
deviations. That is equivalent to saying
the two means are equal to five or the
unknown mean from the sampled popula-
tion equals five. If the statistical
-------
An Initiation Into Statistics
decision was the two populations
disagree then the evidence says that the
mean of the sampled population doesn't
equal five.
In summary, a one sentence nontechnical
definition for a test of hypothesis could
be "A test of hypothesis can be defined
as a method for comparing an unknown
sampled population with a known population
(or mathematical model) and deciding if the
two populations are m agreement (or are
alike) or disagreement (or are not alike). "
A longer nontechnical working definition
for hypothesis testing would be as follows.
A test of significance can be defined as
a method of analyzing data so as to dis-
criminate between two hypotheses. The
first hypothesis is called the null hypothesis
which means there is no difference between
sampled and hypothetical population. The
difference between the two populations is
equal to zero or is null. The second
hypothesis is called the alternative hypoth-
esis which should be the operational
statement of the experimenters research
hypothesis. Next the best test for the
alternative hypothesis is selected, that
is, the one that has the highest probability
of saying the research hypothesis is true
when it is in fact true. The probability of
deciding the null hypothesis is false when
true is selected by the researcher.
This value is called the alpha-error. The
alpha-error specifies those values for the
statistic for which the decision is agreement
(or do not reject HQ or there is no difference
between sampled and hypothetical populations)
or there is disagreement (accept alternative
or research hypothesis). The sample is now
calculated and the statistic value is com-
pared with the decisions values determined
by the alpha-error. The decision is made
to either: 1) do not reject the null hypothesis
or, 2) reject the null hypothesis (or equiva-
lently accept research hypothesis).
For those readers who want an in depth
nontechnical explanation of a tests of
significance, the references listed by this
writer are suggested as a beginning.
Our next topic relationship needs no
introduction since every scientist under-
stands that some sort of predictive
technique is implied.
Variance analysis is next on our list.
In variance analysis, the total variation
displayed by a set of observations may,
in certain circumstances be separated
into components associated with defined
sources of variation. The defined sources
are used as a criterion of classification
for the observations. That portion of
variance which is left undefined or
explained is called experimental error.
Many standard situations can be reduced
to the variance analysis form.
Our ninth topic has many aliases, two
examples are sampling plan and survey
design. The purpose is to measure
nature as it exists by taking a sample
from a population. Statistical data are
collected to provide a rational basis for
action. The action may call for the
enumerative interpretation of the data
or it may call for an analytic interpretation.
These actions determine the difference
between enumerative and analytic surveys.
Enumerative surveys would be classified
in the upper left hand quadrant of Figure 1,
while analytic surveys should appear in the
lower left quadrant of that same figure.
In a design of experiment, our next topic,
the researcher is interested in the change
produced in a response by the influencing
factors. For example, a researcher may
be interested in the yield in bushels of
wheat (the response) when different
fertilizers are used in different amounts
(the influencing factors under his control).
The design of experiment consists of the
set of rules which match treatments (or
list of influencing factors to be studied)
with the units (plots of ground) to which
the treatments are to be applied. The
principles of randomization, replication
and stratification are employed and
produce the many designs available.
In the eleventh topics, the adjective
stochastic implies the presence of a
-------
An Initiation Into Statistics
random variable. Hence a stochastic
process is one wherein the system incor-
porates an element of randomness as
opposed to a deterministic system. A
time series is a set of ordered observa-
tions on a measurable characteristic
taken at different points in time.
The final classification is the miscellaneous
basket for topics which are not large enough
for their own major classification.
VI THIRD VIEWPOINT
The final viewpoint will be the question and
answer approach. This will lead to a more
detailed problem classification viewpoint and
tie in some of the topics discussed in the
second viewpoint.
Suppose you are interested in the B.O. D.
measurements at a particular point in the
Bay of San Francisco. The totality of all
such measurements will be called a parent
population and will be represented by some
distribution as shown in Figure 3.
PARENT
POPULATION
DISTRIBUTION
MEASURABLE CHARACTERISTIC
Figure 3
Three questions the researcher can ask at
this point are:
1) What is the research hypothesis?
Does my boss agree 9
2) What is the definition of our population?
3) Am I going to study my population as
is or subdivide it and study each
subdivision?
Let us assume that the answer to the first
question in our imaginary study has been
finalized. Question one is still the best
starting point in any investigation.
In this imaginary study, the answer to the
second question is clearly answered but in
other studies, the question is not always
easily answered.
Some answers to the third question could
be as follows: I will subdivide the population
and compare daytime versus nighttime. An
alternative choice could be, I will subdivide
into the four seasons and compare.
After these three questions have been satis-
factorily answered, the next question could
be:
4) Will I be able to measure the entire
population or take a sample?
If the answer to this question is: I will
measure all the population, then the data
analysis will be descriptive statistics.
Statistical analysis is subdivided into the
two main areas, namely descriptive and
inferential statistics.
The first of these is descriptive statistics
which is concerned with the problem of how
you condense a large mass of data in such a
way so that you summarize the available
data into several succinct numbers which
is simple to interpret and easier for the mind
to grasp and comprehend. This type of
statistic is the one students first meet in
a course in statistics (See Figure 4).
STATISTICS
DESCRIPTIVE INFERENTIAL
FIGURE 4
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An Initiation Into Statistics
If the answer to question 4 is take a sample
(few investigators have the luxury of
measuring the entire population) then
several other good questions occur like the
following
5) How big a sample do I need' Is my
budget adequate for it'
6) If sample size is not fixed in advance,
how will data collection be terminated9
7) How will our samples be obtained?
Time, place, order, etc.
8) If our samples are not random, how
will we generalize'
Statistical theory assumes random samples,
hence violation of this assumption invalidates
the application of the theory.
9) Am I going to use the sample for
statistical inference with respect to
estimation or hypothesis testing?
(See Figures 5 and 6).
Hypothesis testing can be briefly defined
as a method of analyzing data so as to
discriminate between two hypotheses.
The one hypothesis is the research
hypothesis while the other is generally
the negation of the research hypothesis.
Estimation is concerned with statements
about the numerical value of unknown
population parameters from sample data.
If the estimate is a single number, then
it is called a point estimate. If an interval
is calculated within which the parameter
lies in a probability sense, then it is
called an interval estimate. (See Figure 7)
Hypothesis testing can be subdivided into
two broad types.
If the parent population is normally
distributed, then the hypothesis testing
is said to be parametric. If normality
cannot be proven or safely assumed, then
the test is said to be nonparametric.
(See Figure 7)
USE STATISTICAL INFERENCE
AND THE DATA FROM
PARENT
POPULATION
OR
SAMPLED
POPULATION
THE SAMPLE TO ESTIMATE
PARENT POPULATION
PARAMETERS
DISTRIBUTION
OF THE SAMPL:
RANDOM SAMPLE OF SIZE n
FIGURE 5
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An Initiation Into Statistics
STATISTICS
DESCRIPTIVE
INFERENTIAL
HYPOTHESIS TESTING
FIGURE 6
ESTIM-
ATION
STATISTICS
DESCRIPTIVE
INFERENTIAL
HYPOTHESIS TESTING
ESTIMATION
POINT
INTERVAL
PARAMETRIC NON-
(NORMAL PARAMETRIC
DATA) (NON- NORMAL
DATA)
ONE-
SIDED
TWO-
SIDED
UPPER LIMIT
LOWER LIMIT
FIGURE 7
If hypothesis testing is to be applied, then
some pertinent questions are:
10) What is the null hypothesis?
11) What is the alternative hypothesis?
12) What is the Type I error?
13) What is the Type II error?
14) What is the consequence of a Type I
error ?
15) What is the consequence of a Type II
error?
At this point, other thought provoking
questions can be asked.
16) What are the variables not measured
that affect my observations?
17) Are we going to reject observations
that seem aberrant? If so, on what
basis, objective or subjective?
18) Have I given any thought to lost
information ?
19) Have I given any thought to precision,
accuracy and bias of measurements?
20) If there is more than one observer
involved, how will I insure uniformity
or separate their differences as it
affects my results.
21) How will we minimize our nonsampling
errors ?
Some more thought-provoking questions of
general nature are
22) Will the variable observed be the one
I am really interested in?
23) Is the data analysis predetermined
before data collection or will this be
determined after data collection?
24) What is it we are trying to do ?
25) What do I have to work with9
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An Initiation Into Statistic-3
26) What do I need to know9
27) Who else is concerned or should be 7
I would like to close this section by asking
some pointed questions related to the
computer.
28) Is a computer going to be used or will
the analyses be manual?
29) If a computer is going to be used,
are the programs needed available?
30) If the computer programs are not
available now, will they be ready when
the data has been collected?
31) If the computer is going to be used
and a large mass of data will have to
be entered, how will the data be fed
into the computer?
VII RECOMMENDATIONS
There are many short courses in statistics
available through EPA National Training
Center. Figures 8 and 9 list the 5-day
courses and the 8-hour seminars dealing
with statistics.
FIGURE 8
[
5-DAY
Number Title
801 Basic Environmental Statistics
Environmental Statistics
802
Design of Experiments
804
Nonparametric
806
Analyzing Qualitative Data
810
Applied Regression Analysis
815
Sample Size Determination
820
Survey Sampling
FIGURE 9
8-Hour Seminars
I
'.Number Title
899. 1
Survey Sampling for Managers
899.2
Analysis of Variance and Design
of Experiments
899. 3
Regression and Correlation
899. 4
An Introduction to Hypothesis
Testing
. 899.5
Statistical Quality Control
; 899.6
Estimation and Hypothesis Testing
1
I
for Normal
' 899.7
1
Nonparametric Hypothesis Testing
i 899.8
Introduction to and the Analysis of
i
j
Categorical Data
| 899.9
An Introduction to Probability
The five-day courses are concerned with
applications while the seminars are designed
to acquaint the listeners with the topics and
the type problems that can be solved.
REFERENCES
1 Wold, H. Causal Inference from Observa-
tional Data. J. R. Statist. Soc. A.
119.28.1956.
2 Santner, J. F. An Introduction to Tests of
Significance Training Manual, National
Training Center, DTTB, MDS, OWP.EP
3 Santner, J. F. Variations on a Test of
Significance Training Manual, National
Training Center, DTTB, MDS, OWP.EP
This outline was prepared by Mi. J. 1?. SnnliuM ,
Mathematical Statistician, National Tiannng
Center, MDS, WPO, EPA, Cincinnati, OH
45268.
-------
USING BENTHIC BIOTA IN WATER QUALITY EVALUATIONS
I BENTHOS ARE ORGANISMS GROWING
ON OR ASSOCIATED PRINCIPALLY
WITH THE BOTTOM OF WATERWAYS
Benthos is the noun.
Benthomc, benthal and benthic are adjectives.
II THE BENTHIC COMMUNITY
A Composed of a wide variety of life
forms that are related because they
occupy "common ground"--the water-
ways bottom substrates. Usually
they are attached or have relatively
weak powers of locomotion. These
life forms are
1 Bacteria
A wide variety of decomposers
work on organic materials,
breaking them down to elemental
or simple compounds (hetero-
trophic). Other forms grow on
basic nutrient compounds or form
more complex chemical compounds
(autotrophic).
2 Algae
Single-cell plants that are the
basic producers of food that
nurtures the animal components
of the community.
3 Flowering Aquatic Plants(Pondweeds)
The largest flora, composed of
complex and differentiated tissues.
Many are rooted.
4 Micro-Fauna
Animals that pass through a U. S.
Standard Series No. 30 sieve, but
are retained on a No. 100 sieve.
Examples are rotifers and micro-
crustaceans. Some forms have
organs for attachment to substrates.
while others burrow into soft
materials or occupy the inter-
stices between rocks, floral
or faunal materials.
5 Macro-Fauna
Animals that are retained on a
No. 30 sieve. This group in-
cludes the insects, worms,
molluscs, and occasionally fish.
Fish are not normally considered
as benthos, though there are
bottom dwellers such as sculpins
and darters.
B It is a self-contained community, though
there is interchange with other commun-
ities. For example: Plankton settles
to it, fish prey on it and lay their eggs
there, terrestrial detritus is added to it,
and many aquatic insects migrate from
it to the terrestrial environment for
their mating cycles.
C It is a stationary water quality monitor
The low motility of the biotic compon-
ents requires that they "live with" the
quality changes of the over-passing
waters. Changes imposed in the long-
lived components remain visible for
extended periods, even after the cause
has been eliminated. Only time will
allow a cure for the community by
migration and reproduction
III HISTORY OF BENTHIC OBSERVATIONS
A Ancient literature records the vermin
associated with fouled waters.
B 500-year-old fishing literature refers
to animal forms that are fish food and
used as bait.
C The scientific literature associating
biota to water pollution problems is
over 100 years old (Mackenthun and
Ingram, 1964).
BI.MET.fm. 8d. 9.72
-------
Using Benthic Biota 111 Water Quality Evaluations
D Early this century, applied biological
investigations were initiated.
1 The entrance of State boards of Health
into water pollution control activities.
2 Creation of state conservation agencies.
3 Industrialization and urbanization.
4 Growth of limnological programs
at universities.
E A decided increase in benthic studies
occurred in the 1950 decade, and much
of today's activities are strongly influenced
by developmental work conducted during
this period. Some of the reasons for this
are:
1 Movement of the universities from
"academic biology" to applied
pollution programs.
2 Entrance of the federal government
into enforcement aspects of water
pollution control.
3 A rising economy and the development
of federal grant systems.
4 Environmental Protection Programs
are a current stimulus.
IV WHY THE BENTHOS''
A It is a natural monitor
B The community contains all of the components
of an ecosystem.
1 Reducers
2 Producers
3 Consumers
a Herbivores
b Predators
C Economy of Survey
1 Manpower
2 Time
3 Equipment
D Extensive Supporting Literature
V REACTIONS OF THE COMMUNITY TO
POLLUTANTS
A Destruction of Organism Types
1 Beginning with the most sensitive forms,
pollutants kill m order of sensitivity
until the most tolerant form is the last
survivor. This results in a reduction
of variety or diversity of organisms.
2 The usual order of macroinvertebrate
disappearance on a sensitivity scale
below pollution sources is shown in
Figure 2.
Stoneflies
Mayflies
Caddisflies
Amphipods
Isopods
Midges
Oligochaetes
As water quality improves, these
reappear in the same order.
B The Number of Survivors Increase
1 Competition and predation are reduced
between forms.
2 When the pollutant is a food (plants,
fertilizers, animals, organic materials).
C The Number of Survivors Decrease
1 The material added is toxic or has no
food value.
2 The material added produces toxic
conditions as a byproduct of decom -
position (e.g., large organic loadings
produce an anaerobic environment
resulting in the production of toxic
sulfides, methanes, etc.)
-------
Using Benthic Biota in Water Quality Evaluations
D The Effects May be Manifest m Combinations
1 Of pollutants and their effects.
2 Vary with longitudinal distribution in
a stream. (Figure 1)
DIRECTION OF FLOW ¦
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TIME OR DISTANCE
..NUMBER OF KINDS
NUMBER OF ORGANISMS
SLUDGE DEPOSITS
Four basic responses of bottom animals to pollution
A Organic wastes eliminate the sensitive bottom animals
and provide food in the form of sludges for the surviving toler-
ant forms B Large quantities of decomposing organic wastes
eliminate sensitive bottom animals and the excessive quanti-
ties of byproducts of organic decomposition inhibit the tolerant
forms; in time, with natural stream purification, water quality
improves so that the tolerant forms can flourish, utilizing the
sludges as food C. Toxic materials eliminate the sensitive
bottom animals, sludge is absent and food is restricted to that
naturally occurring m the stream, which limits the number of
tolerant surviving forms Very tome materials may eliminate
all organisms below a waste source D. Organic sludges with
toxic materials reduce the number of kinds by eliminating
sensitive forms Tolerant survivors do not utilize the organic
sludges because the toxicity restricts their growth
Figure 1
E Tolerance Grouping (Figure 2)
Flexibility must be maintained in the
establishment of tolerance lists based
on the response of organisms to the
environment because of complex relation-
ships among varying environmental
conditions. Some general tolerance
patterns can be established. Stonefly
nymphs, mayfly naiads, hellgrammites,
and caddisfly larvae represent a grouping
(sensitive or intolerant) that is quite
sensitive to environmental changes.
Blackfly larvae, scuds, sowbugs, snails,
fingernail clams, dragonfly nymphs,
damselfly nymphs, and most kinds of
midge larvae are intermediate (facultative
or intermediate) in tolerance. Sludge-
worms, some kinds of midge larvae
(bloodworms), and some leeches are
tolerant to comparatively heavy loads
of organic pollutants. Sewage mosquitoes
and rat-tailed maggots are tolerant of
anaerobic environments.
F Structural Limitations
The morphological structure of a species
limits the type of environment it may
occupy. Species with complex appendages
and exposed, complicated respiratory
structures, such as stonefly nymphs,
mayfly numphs, and caddisfly larvae,
that are subjected to a constant deluge
of settleable particulate matter soon
abandon the polluted area because of the
constant preening required to mamtain
mobility or respiratory functions, other-
wise, they are soon smothered.
Species without complicated external
structures, such as bloodworms and
sludgeworms, are not so limited in
adaptability. A sludgeworm, for example,
can burrow in a deluge of particulate organic
matter and flourish on the abundance of
"manna. " Morphology also determines
the species that are found in riffles, on
vegetation, on the bottom of pools, or in
bottom deposits.
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Using Benthic Biota in Water Quality Evaluations
VI SAMPLING PROCEDURES
A Fauna
1 Qualitative sampling determines the
variety of species occupying an area.
Samples may be taken by any method
that will capture representatives of the
species present. Collections from such
samplings indicate changes in the
environment, but generally do not
accurately reflect the degree of
change. Mayflies, for example, may
be reduced from 100 to 1 per square
foot. Qualitative data would indicate
the presence of both species, but might
not necessarily delineate the change in
predominance from mayflies to sludge-
worms.
2 Quantitative sampling is performed to
observe changes in predominance. The
most common quantitative sampling
tools are the Petersen and Ekman
dredges and the Surber stream bottom
or square-foot sampler. Of these, the
Petersen dredge samples the widest
variety of substrates. The Ekman
dredge is limited to fine-textured and
soft substrates, such as silt and sludge.
The Surber sampler is designed for
sampling riffle areas; it requires
moving water to transport dislodged
organisms into its net and is limited
to depths of two feet or less.
3 The collected sample is screened with
a standard sieve to concentrate the
organisms, these are sorted from the
retained material, and the number of
each kind determined. Data are then
adjusted to number per unit area,
usually to the number per square foot
of bottom or occasionally to number
per square meter. This adjustment
standardized the method of data
expression.
4 Independently, neither qualitative nor
quantitative data suffice for thorough
analyses of environmental conditions.
A cursory examination to detect damage
may be made with either method, but
a combination of the two gives a more
precise determination. If a choice must
be made, quantitative sampling would
be best, because it incorporates a
partial qualitative sample.
REPRESENTATIVE BOTTOM-DWELLING MACROANIMALS
Drawings from Geckler, j.( K. M. Mackenthun and W. M. Ingram, 1963.
Glossary of Commonly Used Biological and Related Terms m Water and
Wastewater Control, DHEW, PHS, Cincinnati, Ohio, Pub. No. 999-WP-2.
A
Stonefly nymph
(Plecoptera)
I
Fingernail clam
(Sphaeriidae)
B
Mayfly nymph
(Ephem e ropt e ra)
J
Damselfly nymph
(Zygoptera)
C
Hellgrammite or
K
Dragonfly nymph
(Amsoptera)
Dobsonfly larvae
(Megaloptera)
L
Bloodworm or midge
D
Caddisfly larvae
(Trichoptera)
fly larvae
(Chironomidae)
E
Black fly larvae
(Simuliidae)
M
Leech
(Hirudinea)
F
Scud
(Amphipoda)
N
Sludgeworm
(Tubificidae)
G
Aquatic sowbug
(Isopoda)
O
Sewage fly larvae
(Psychodidae)
H
Snail
(Gastropoda)
P
Rat bailed maggot
(Tubifera-Eristalis)
KEY TO FIGURE 2
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Using Benthic Biota in Water Quality Evaluations
B ^ C
SENSITIVE
¦- A
E
F G
INTERMEDIATE
IV' m
TOLERANT
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Using Benthic Biota in Water Quality Evaluations
B Flora
1 Direct quantitative sampling of natu-
rally growing bottom algae is difficult.
It is basically one of collecting algae
from a standard or uniform area of the
bottom substrates without disturbing
the delicate growths and thereby dis-
tort the sample. Indirect quantitative
sampling is the best available method.
Artificial substrates, such as wood
blocks, glass or plexiglass slides,
bricks, etc., are placed in a stream.
Bottom-attached algae will grow on
these artificial substrates. After two
or more weeks, the artificial sub-
strates are removed for analysis.
Algal growths are scraped from the
substrates and the quantity measured.
Since the exposed substrate area and
exposure periods are equal at all of
the sampling sites, differences in the
quantity of algae can be related to
changes in the quality of water flowing
over the substrates.
2 The quantity of algae on artificial sub-
strates can be measured in several
ways. Microscopic counts of algal
cells (Cooke, 1958, Gumtow, 1955)
and dry weight of algal material
(Castenholz, 1960, Grezenda and
Brehmer, 1960) are long established
methods.
Microscopic counts involve thorough
scraping, mixing, and suspension of
the algal cells. From this mixture
an aliquot of cells is withdrawn for
enumeration under a microscope. Dry
weight is determined by drying and
weighing the algal sample, then ig-
niting the sample to burn off the algal
materials, leaving inert inorganic
materials that are again weighed.
The difference between initial weight
and weight after ignition is attributed
to algae.
Any organic sediments, however, that
settle on the artificial substrate along
with the algae are processed also.
Thus, if organic wastes are present
appreciable errors may enter into
this method.
3 During the past decade, chlorophyll
analysis has become a popular method
for estimating algal growth. Chloro-
phyll is extracted from the algae and
is used as an index of the quantity of
algae present. The advantages of
chlorophyll analysis are rapidity,
simplicity, and vivid pictorial results.
The algae are scrubbed from the
artificial substrate samples, filtered
to remove excess water, then each
sample is placed (with filter) in equal
volumes of acetone or alcohol, which
extracts the chlorophyll from the algal
cells. A second filtering to remove
the extracted algae and other detritus
produces a non-turbid, colored chlor-
ophyl extract. The chlorophyll ex-
tracts may be compared visually.
Because the chlorophyll extracts fade
with time, colorimetry shouldbeused
for permanent records. Harvey (1934)
describes an artificial color standard.
Richards with Thompson (1952) de-
scribes a method of colorimetry that
determines the quantities of various
kinds of chlorophyll along with other
plant and animal pigments. For rour
tine records, simple colorimeters
will suffice. At very high chlorophyll
densities, interference with colori-
metry occurs, which must be corrected
through serial dilution of the sample
or with a nomograph
VII FACTORS INVOLVED IN DATA INTER-
PRETATION
Two very important factors in data evalua-
tion are a thorough' knowledge of conditions
under which the data were collected and a
critical assessment of the reliability of the
data's representation of the situation.
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Using Benthic Biota in Water Quality Evaluations
A Maximum-Minimum Values
The evaluation of physical and chemical
data to determine their effects on aquatic
organisms is primarily dependent on
maximum and minimum observed values.
The mean is useful only when the data are
relatively uniform. The minimum or
maximum values usually create acute
conditions in the environment.
B Identification
Precise identification of organisms to
species requires a specialist in limited
taxonomic groups. Many immature
aquatic forms have not been associated
with the adult species. Therefore, one
who is certain of the genus but not the
species should utilize the generic name,
not a potentially incorrect species name.
The method of interpreting biological
data on the basis of numbers of kinds
and numbers of organisms will typically
suffice.
C Lake and Stream Influence
Physical characteristics of a body of
water also affect animal populations.
Lakes or impounded bodies of water
support different faunal associations
from rivers. The number of kinds
present in a lake may be less than that
found in a stream because of a more
uniform habitat. A lake is all pool,
but a river is composed of both pools
and riffles. The nonflowing water of
a lake exhibits a more complete set-
tling of particulate organic matter that
naturally supports a higher population
of detritus consumers. For these
reasons, the bottom fauna of a lake
or impoundment cannot be directly
compared with that of a flowing stream.
D Extrapolation
How can bottom-dwelling macrofauna
data be extrapolated to other environ-
mental components' It must be borne
in mind that a component of the total
environment is being sampled. If the
sampled component exhibits changes,
then so must the other interdependent
components of the environment. For
example, a clean stream with a wide
variety of desirable bottom organisms
would be expected to have a wide vari-
ety of desirable bottom fishes; when
pollution reduces the number of bottom
organisms, a comparable reduction
would be expected in the number of
fishes. Moreover, it would be logical
to conclude that any factor that elim-
inates all bottom organisms would
eliminate most other aquatic forms
of life.
VIII IMPORTANT ASSOCIATED ANALYSES
A The Chemical Environment
1 Dissolved oxygen
2 Nutrients
3 Toxic materials
4 Acidity and alkalinity
5 Etc
B The Physical Environment
1 Suspended solids
2 Temperature
3 Light penetration
4 Sediment composition
5 Etc
IX AREAS IN WHICH BENTHIC STUDIES
CAN BEST BE APPLIED
A Damage Assessment
If a stream is suffering from pollutants,
the biota will so indicate A biologist
can determine damages by looking at the
"critter" assemblage in a matter of hours
Usually, if damages are not found, it will
not be necessary to alert the remainder
of the agency's staff, pack all the equip-
ment, pay travel and per diem, and then
wait five days before enough data can be
assembled to begin evaluation.
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I"sing Bomhio Biota 111 Walei Quality Evaluations
B By determining what damages have been
' done, the potential cause "list" can be
reduced to a few items for emphasis and
the entire "wonderful worlds" of science
and engineering need not be practiced with
the result that much data are discarded
later because they were not applicable to
the problem being investigated
C Good benthic data coupled with chemical,
physical and engineering materials can be
used to predict the direction of future
changes and to estimate the amount of
pollutants that need to be removed from
the waterways.
REFERENCES
1 Castenholz, R.W. Season Changes in the
Attached Algae of Freshwater and Saline
Lakes in the Lower Grand Coulee,
Washington. Limnology and Oceanography
5 (1): 1 1960.
2 Cooke, W.B. Continuous Sampling of
Trickling Filter Populations I.
Procedures. Sewage and Industrial
Wastes 30( 1):21. 1958.
3 Grezenda, A.R. and Brehmer, M. L.
A Quantitative Method for Collection
and Measurement of Stream Periphyton.
Limnology and Oceanography 5(2)rl90.
I960.
7 Keup, L. E., Ingram, W.M and
Mackenthun, K. M The Role of
Bottom Dwelling Macrofauna in Water
Pollution Investigations. USPHS
Environmental Health Series Publ.
No 999-WP-38, 23 pp. 1966.
8 Keup, L. E., Ingram, W.M. and
Mackenthun, K.M. Biology of Water
Population- A Collection of Selected
Papers on Stream Pollution, Waste
Water, and Water Treatment.
Federal Water Pollution Control
Administration Pub No. CWA-3,
290 pp. 1967.
9 Mackenthun, K. M. The Practice of
Water Pollution Biology FWQA.
281 pp. 1969.
10 Richards, F.A. and Thompson, T. G.
The Estimation and Characterization
of Plankton Populations by Pigment
Analysis II. A Spectrophotometric
Method for the Estimation of Plankton
Pigments. Journal Marine Research
11(2)-156. 1952.
11 Stewart, R.K , Ingram, W.M and
Mackenthun, K.M. Water Pollution
Control, Waste Treatment and Water
Treatment: Selected Biological Ref-
erences on Fresh and Marine Waters.
FWPCA Pub. No. WP-23, 126 pp.
1966
4 Gumtow, R.B. An Investigation of the
Periphyton in a Riffle of the West
Gallatin River, Montana. Trans. Am.
Microscopical Society 74(3)-278. 1955
5 Harvey, H.W. Measurement of Phytoplankton
Populationb. Journal Marine Biological
Assoc 19-701 1934.
6 Hynes>, H B.N. The Ecology of Running
Waters. Univ. Toronto Press. 1970.
This outline was prepared by Lowell E. Keup,
Chief, Technical Studies Branoh, Div, of
Technical Support, EPA, Washington, D. C.
20242.
-------
CASE PREPARATION AND COURTROOM PROCEDURE
I TYPES OF PROCEEDINGS IN WHICH
WATER QUALITY EVIDENCE MAY
BE USED
A Administrative Proceedings
1 Rule making
a Setting up of regulations having
general application, e.g., stream
classifications and implementation
plan target dates
b Factors of safety and absolute
prohibitions may be appropriate
2 Adjudications
a Determinations by agency having
expertise with respect to particular
discharge or discharger, e.g.,
approval of plans and specs and
time schedule of a particular
discharger
B Court Actions
1 Civil in behalf of state or federal
government
a Actions to compel action or sus-
pension of action - nuisance, health
hazard, etc.,--including court
action following federal conference
--hearing procedure
b Discharges from specific industries
c Littering
d Discharges harmful to fish and/or
crustaceans
e Discharges harmful to specific
types of receiving waters
f Discharges of poisons
NOTE--In some of these situations
doing the act may constitute
the violation, in others
proof of intent or knowledge
of effects may also have to
be proved.
3 Private actions for damages or
to compel action
a Alleged harm to plaintiff, e.g.,
pollution of stream killing animals
C Procedural Matters
1 See Attached sheet "Administrative
and Court Proceedings" on Burden
of proof, fact finding, and methods of
presentation of evidence.
D Classes of Evidence - General Rules
1 Facts - direct
b Violations of Water Quality Standards
c Violations of Effluent Standards or
discharge permits
d Tort or contract actions relating to
design and/or operation of treatment
facilities
2 Criminal (dependent on content of
applicable statutes)
a Discharge of specific materials
a The material was floating from
the outfall.
2 Derived values - expert testimony
- test results and/or opinion as to
effects
a The D. O. was zero, the waterway
was polluted, the plant can be built
in 6 months.
3 Hearsay
a Joe told me
W.Q. le. la. 1. 72
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Case Preparation and Courtroom Procedure
4 Relevancy
5 Admissibility vs. weight
a Even if admissible, the weight to be
given is up to fact finder-- credi-
bility.
E Admissibility of Results of Sampling
and Testing (Numbers)
1 Sampling
a Cham of custody
b Tags, etc.
c Containers
d Place and time
e Retention of samples (Proving that
the sample represents what is at
issue in the action (relevancy),that
there has been no opportunity for
tampering, and availability of
portions for analysis by other side
(non-transitory criteria) ).
2 Analysis
a Who performed (Can identity of
each participant be shown')
b Admission through supervisor
custodian
c Scientific acceptance of method.
Is there a particular method required
to be used by the agency9
d Propriety of conduct
e Retention of bench cards and other
indicia of results. (Your attorney
can make arrangements to substitute
copies for originals).
3 Tests
a Comparison with actual conditions
b Mathematical models - how can a
computer be cross-examined9
F Admissibility of Expert Opinion on
Causes and Effects
1 Who has special knowledge - and of
what particular areas7
2 Indicators
3 Significance of numerical determin-
ations or observations
4 Consistency with own prior publications
and testimony
5 Have underlying facts been or need to
be proved--first hand information of
this and/or comparable situations.
6 Use of treatises
G Conduct on the Witness Stand
1 General
a On direct - know what counsel will
ask and let him know generally
what you will answer, but don't
make it sound rehearsed.
b Use layman's language to extent
possible.
c Listen to question and answer it
to best of your ability.
d Speak so that court reporter, judge,
jury, and counsel can hear you.
e Speak in language that will be
understood, don't talk down.
f Answer only what you are asked
--don't volunteer, however, answer
with precision.
g There is nothing wrong with asking
to have a question repeated or
rephrased.
h There is nothing wrong with saying
that you consulted with your
attorney before you testified, but
beware of the question "Did Mr. X
tell you what to say9 "
-------
Case Preparation and Courtroom Procedure
1 There is nothing wrong with thinking
out your answer before responding.
j You are not expected to know all
the answers—if you do not know,
admit it.
k Don't attempt to answer questions
outside your area of personal
knowledge (hearsay) or beyond your
expertise. (Your may be an expert
on conducting laboratory tests, but
not on epidimeological inferences
from results).
1 Don't try to answer before the judge
rules on objection.
m Show that you are an impartial
dispenser of information and/or
opinion, not a protagonist.
n Don't be afraid to admit what may
appear to be damaging.
2 If you are testifying as an expert
a Establish qualifications -- give
information relevant to your area
of expertise -- educational (in-
cluding this course'), work,
publications, number of times you
have testified previously.
b Differentiate between physical facts
(measurements and observations)
and opinion (derived values).
c Be prepared to discuss theory (in-
cluding assumptions) instruments
used, techniques(including choice
of a particular technique),physical
limitations and errors, inter-
ferences
d If experiments were conducted,
be able to justify both as to theory
and relevancy to this litigation.
e If you're being paid to testify,
admit it.
3 Scientific personnel as advisers to
counsel
a Review and refamiliarize self with
materials before you discuss with
your attorney.
b Be in a position to present all facts
known to you simply and concisely
Who, What, When, Where, and
Why, How.
c Don't overlook facts and/or test
results because you don't think
they're important. Let attorney
decide what he needs.
d Use of standard report forms
e Ability to recommend additional
witnesses with needed specialized
knowledge
f Ability to aid in cross-examination
of other side's experts and reconcile
opinions and/or results
g Be candid - sometimes better not
to start a lawsuit or accept a
settlement than lose in the end.
H Non-Verbal Presentation of Evidence
1 Exhibits - including photographs
2 Summaries
3 Business and/or government records
a Prepared contemporaneously and
m usual course of activities
4 Pre-prepared direct examination
a Usually limited to actions before
ICC, FPC, and other federal
agencies.
I Criminal Procedure
1 Privilege Against Self Incrimination
(available only to persons)
a Warning and suspects
b Effect of duty to report spills
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Case Preparation and Courtroom Procedure
c Effect of duty to obtain license or
permit and/or furnish operating
reports
3 Unreasonable search and seizure
a Available to persons and
corporations
d Immunity from prosecution
4 Procedures and need for arrest and
This outline was prepared by David I. Shedroff,
Enforcement Analyst, Office of Enforcement
and General Counsel, Cincinnati Field
Investigations Center, 5555 Ridge Avenue,
Cincinnati, OH 45268.
Administrative &. Court Proceedings,
and Excerpts from Revised Draft of
Proposed Rules of Evidence for the
United States Courts can be found on the
following pages.
2 Double Jeopardy
search warrants -- possible cause
-------
ADMINISTRATIVE & COURT PROCEEDINGS
Court or Agency
Fact Finder
Burden of Proof
Comments
State Pollution Control
Agency
Rule making-adjudi-
cation
Federal Water Pollution
Control Act
Conference
Hearing
Court
Court
Civil Case --
- for money only
- injunction
preliminary or
temporary
permanent
- administrative
appeal
Agency
Head of agency
Hearing Board
Judge
Judge or jury
Judge
Judge
Judge - whether
"arbitrary and
capricious" or sub-
stantial evidence
As per statute -
usually weight of
evidence.
Weight of evidence
Must show immediate
harm or danger.
Usually clear and convincing.
Hearing may be conducted by hearing
examiner, agency member, or full
agency. Appeal may be on facts and
law or law alone, depending on statute.
Reports acceptable.
Specific testimony.
Uses prior material, and may take
additional testimony.
Must also show likelihood of success at
final hearing - bond required for non-
government plaintiff.
"Balance Equities"
Sometimes have complete new trial.
Criminal case
includes penalties
Jury unless waived. Beyond reasonable
doubt.
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Case Preparation and Courtroom Procedure
Excerpts from Revised Draft of Proposed
RULES OF EVIDENCE FOR THE UNITED STATES COURTS
GENERAL PROCEDURES
Rule 102.
PURPOSE AND CONSTRUCTION
These rules shall be construed to secure fairness in administration, elimination
of unjustifiable expense and delay, and promotion of growth and development of
the law of evidence to the end that the truth may be ascertained and proceedings
justly determined.
Rule 101.
PRELIMINARY QUESTIONS
(a) Questions of Admissibility Generally. Preliminary questions concerning the
qualification of a person to be a witness, the existence of a privilege, or the
admissibility of evidence shall be determined by the judge, subject to the pro-
visions of subdivision (b). In making his determination he is not bound by the
rules of evidence except those with respect to privileges.
(b) Relevancy Conditioned on Fact. When the relevancy of evidence depends upon
the fulfillment of a condition of fact, the judge shall admit it upon, or subject to,
the introduction of evidence sufficient to support a finding of the fulfillment of the
condition.
Rule 615.
EXCLUSION OF WITNESSES
At the request of a party the judge shall order witnesses excluded so that they
cannot hear the testimony of other witnesses, and he may make the order of his
own motion. This rule does not authorize exclusion of (1) a party who is a natural
person, or (2) an officer or employee of a party which is not a natural person
designated as its representative by its attorney, or (3) a person whose presence
is shown by a party to be essential to the presentation of his cause.
Rule 611.
MODE AND ORDER OF INTERROGATION AND PRESENTATION
(a) Control by Judge. The judge may exercise reasonable control over the mode
and order of interrogating witnesses and presenting evidence so as to (1) make the
interrogation and presentation effective for the ascertainment of the truth, (2) avoid
needless consumption of time, and (3) protect witnesses from harassment or undue
embarrassment.
(b) Scope of Cross-Examination. A witness may be cross-examined on any matter
relevant to any issue in the case, including credibility. In the interests of justice,
the judge may limit cross-examination with respect to matters not testified to on
direct examination.
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Case Preparation and Courtroom Procedure
Rule 613.
PRIOR STATEMENTS OF WITNESSES
(a) Examining Witness Concerning Prior Statement. In examining a witness
concerning a prior statement made by him, whether written or not, the state-
ment need not be shown or its contents disclosed to him at that time, but on
request the same shall be shown or disclosed to opposing counsel.
JUDICIAL NOTICE
Rule 201.
JUDICIAL NOTICE OF ADJUDICATIVE FACTS
(b) Kinds of Facts. A judicially noticed fact must be one not subject to reasonable
dispute in that it is either (1) generally known within the territorial jurisdiction of
the trial court or (2) capable of accurate and ready determination by resort to
sources whose accuracy cannot reasonably be questioned.
(g) Instructing Jury. The judge shall instruct the jury to accept as established
any facts judicially noticed.
RELEVANCE
Rule 401.
DEFINITION OF "RELEVANT EVIDENCE"
"Relevant evidence" means evidence having any tendency to make the existence
of any fact that is of consequence to the determination of the action more probable
or less probable than it would be without the evidence.
Rule 402.
RELEVANT EVIDENCE GENERALLY ADMISSIBLE,
IRRELEVANT EVIDENCE INADMISSIBLE
All relevant evidence is admissible, except as otherwise provided by these rules,
by other rules adopted by the Supreme Court, by Act of Congress, or by the
Constitution of the United States. Evidence which is not relevant is not admissible.
COMPETENCY OF WITNESSES
Rule 601.
GENERAL RULE OF COMPETENCY
Every person is competent to be a witness except as otherwise provided in these
rules.
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Case Preparation and Courtroom Procedure
Rule 602.
LACK OF PERSONAL KNOWLEDGE
A witness may not testify to a matter unless evidence is introduced sufficient
to support a finding that he has personal knowledge of the matter. Evidence to
prove personal knowledge may, but need not, consist of the testimony of the
witness himself. This rule is subject to the provisions of Rule 703, relating to
opinion testimony by expert witnesses.
EXPERT TESTIMONY
Rule 702.
TESTIMONY BY EXPERTS
If scientific, technical, or other specialized knowledge will assist the trier of
fact to understand the evidence or to determine a fact in issue, a witness qualified
as an expert by knowledge, skill, experience, training, or education, may testify
thereto in the form of an opinion or otherwise.
Rule 703.
BASES OF OPINION TESTIMONY BY EXPERTS
The facts or data in the particular case upon which an expert bases an opinion or
inference may be those perceived by or made known to him at or before the hearing.
If of a type reasonably relied upon by experts in the particular field in forming
opinions or inferences upon the subject, the facts or data need not be admissible
in evidence.
Rule 705.
DISCLOSURE OF FACTS OR DATA UNDERLYING EXPERT OPINION
The expert may testify in terms of opinion or inference and give his reasons
therefore without prior disclosure of the underlying facts or data, unless the
judge requires otherwise. The expert may in any event be required to disclose
the underlying facts or data on cross-examination.
Rule 706.
COURT APPOINTED EXPERTS
(a) Appointment. The judge may on his own motion or on the motion of any
party enter an order to show cause why expert witnesses should not be appointed,
and may request the parties to submit nominations. The judge may appoint any
expert witnesses agreed upon by the parties, and may appoint witnesses of his
own selection. An expert witness shall not be appointed by the judge unless he
consents to act. A witness so appointed shall be informed of his duties by the
judge in writing, a copy of which shall be filed with the clerk, or at a conference
in which the parties shall have opportunity to participate. A witness so appointed
shall advise the parties of his findings, if any, his deposition may be taken by any
party, and he may be called to testify by the judge or any party. He shall be subject
to cross-examination by each party, including a party calling him as a witness.
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Case Preparation and Courtroom Procedure
HEARSAY
Rule 801.
DEFINITIONS
The following definitions apply under this Article:
(a) Statement. A "statement" is (1) an oral or written assertion or
(2) nonverbal conduct of a person, if it is intended by him as an assertion.
(b) Declarant. A "declarant" is a person who makes a statement.
(c) Hearsay. "Hearsay" is a statement, other than one made by the declarant
while testifying at the trial or hearing, offered in evidence to prove the truth
of the matter asserted.
Rule 802.
HEARSAY RULE
Hearsay is not admissible except as provided by these rules or by other rules
adopted by the Supreme Court or by Act of Congress.
Rule 803.
HEARSAY EXCEPTIONS: AVAILABILITY OF DECLARANT IMMATERIAL
The following are not excluded by the hearsay rule, even though the declarant is
available as a witness
(5) Recorded Recollection. A memorandum or record concerning a matter about
which a witness once had knowledge but now has insufficient recollection to enable
him to testify fully and accurately, shown to have been made when the matter was
fresh in his memory and to reflect that knowledge correctly. If admitted, the
memorandum or record may be read into evidence but may not itself be received
as an exhibit unless offered by an adverse party.
(6) Records of Regularly Conducted Activity. A memorandum, report, record,
or data compilation, in any form, of acts, events, conditions, opinions, or
diagnoses, made at or near the time by, or from information transmitted by,
a person with knowledge, all in the course of a regularly conducted activity, as
shown by the testimony of the custodian or other qualified witness, unless the
sources of information or other circumstances indicate lack of trustworthiness.
(18) Learned Treatises. To the extent called to the attention of an expert witness
upon cross-examination or relied upon by him m direct examination, statements
contained in published treatises, periodicals, or pamphlets on a subject of
history, medicine, or other science or art, established as a reliable authority
by the testimony or admission of the witness or by other expert testimony or by
judicial notice. If admitted, the statements may be read into evidence but may
not be received as exhibits.
-------
Case Preparation and Courtroom Procedure
IDENTIFICATION OF PERSONS AND SAMPLES
Rule 901.
REQUIREMENT OF AUTHENTICATION OR IDENTIFICATION
(a) Gensral Provision The requirement of authentication or identification as a
condition precedent to admissibility is satisfied by evidence sufficient to support
a finding that the matter m question is what its proponent claims.
(b) Illustrations. By way of illustration only, and not by way of limitation, the
following are examples of authentication or identification conforming with the
requirements of this rule
(1) Testimony of Witness with Knowledge. Testimony that a matter is what it is
claimed to be.
(3) Comparison by Trier or Expert Witness. Comparison by the trier of fact or
by expert witnesses with specimens which have been authenticated
(9) Process or System. Evidence describing a process or system used to produce
a result and showing that the process or system produces accurate result.
ADMISSIBILITY AND PROOF OF SPECIAL MATTERS
Rule 406.
HABIT, ROUTINE PRACTICE
(a) Admissibility. Evidence of the habit of a person or of the routine practice of an
organization, whether corroborated or not and regardless of the presence of eye-
witnesses, is relevant to prove that the conduct of the person or organization on a
particular occasion was in conformity with the habit or routine practice.
(b) Method of Proof. Habit or routine practice may be proved by testimony in the
form of an opinion or by specific instances of conduct sufficient in number to warrant
a finding that the habit existed or that the practice was routine.
Rule 612
WRITING USED TO REFRESH MEMORY
If a witness uses a writing to refresh his memory, either before or while testifying,
an adverse party is entitled to have it produced at the hearing, to inspect it, to
cross-examine the witness thereon, and to introduce in evidence those portions which
relate to the testimony of the witness.
Rule 1006.
SUMMARIES
The contents of voluminous writings, recordings, or photographs which cannot
conveniently be examined in court may be presented in the form of a chart, summary,
or calculation. The originals, or duplicates, shall be made available for examination
or copying, or both, by other>parties at a reasonable time and place The judge may
order that they be produced in court.
-------
BACTERIA AND PROTOZOA
AS TOXICOLOGICAL INDICATORS
I INTRODUCTION
The WINOGRADSKY COLUMN is an excellent
simple classroom experiment as well as a
miniature ecosystem which yields a variety
of photosynthetic and other protista especially
the bacterial forms important in photosynthesis
research. These photosynthetic bacteria, as
pointed out by Dr. Hutner, are ubiquitous in
wet soils and natural waters but ordinarily
escape notice.
H PREPARING THE COLUMN
The materials needed are simple laboratory
items.
A The inoculum (a black sludge) may be
easily obtained from a local sewage
treatment plant or the bottom of a pond
or lake. Because the USPHS document,
containing Dr. Hutner's paper, is out of
print it is reproduced here.
B Directions for preparing the column and
other useful information are given in that
paper.
C Dr. Hutner's bibliography should be
sufficient for those who wish more
information.
Ill ECOSYSTEM DEVELOPMENT
A Factors such as the substrate used, the
inoculum, the overlying supernatant water,
and laboratory conditions as temperature
and light, will all influence the particular
type of biota forming successive layers
or zones. The accompanying figure is
therefore generalized and is not intended
to be absolute.
B Some representative forms are listed for
general information. The numbers
correspond to those on the figure.
1 Inorganic substrate on toweling.
2 Green photosynthetic bacteria
Microchloris, Chlorobium, and
Chlorochromatum, methane bacteria,
and SO., reducers.
4
3 Photosynthetic purple sulfur bacteria.
Thiopedia and Thiosarcma.
4 Filamentous sulfur bacteria,
Beggiatoa.
5 Non-sulfur photosynthetic bacteria,
Rhodopseudomonas.
6 Blue-green algae, Schizothrix and
Oscillatoria.
7 Diatoms, Nitzschia.
8 Coccoid green algae, Ankistrodesmus,
and flagellate greens, Chlamydomonas,
similar to a stabilization pond flora.
9 Filamentous green algae, Ulothrix.
C Besides the photosynthetic bacteria and
other protista there will be a variety of
protozoa found in the aerobic levels
(app. 6 through 9). Many of these
protozoans are typical fauna of activated
sludge.
D The possibilities are endless for further
experimentation. These ecosystems are
also convenient and inexpensive sources
for protozoa and other protista for class
and laboratory instruction.
IV MICRO AQUARIA
A Fenchel describes a micro aquarium
(1.5 X5 cm) which may be observed under
a compound microscope. (Figure 2)
B The development of communities of
organisms is quite similar to the
Winogradsky Column. (Figure 3)
PC.20a 9.72
-------
Bacteria and Protozoa as Toxicological Indicators
C The basic media consists of one liter of
natural water 10 g CaSO^, 1 g glucose,
1 g of peptone, autoclaved and stored at
5° C.
D Before use agar is boiled with the media.
While hot, the media is introduced into
one end of the "micro aquarium" with a
pipet. After the agar congeals, natural
water samples are added. During
incubation and when not being observed,
the micro aquaria are kept in a moist
chamber.
3 Burbanck, W.D. and Spoon, D.M.
The Use of Sessile Ciliates Collected
in Plastic Petri Dishes for Rapid
Assessment of Water Pollution.
Jour, of Protozoology. 14(4)-739-744.
1967.
4 Curds, C.R. and Cockburn, A.
Protozoa in Biological Sewage Treat-
ment Processes. I. A Survey of the
Protozoan Fauna of British Percolating
Filters and Activated Sludge Plants,
Water Research 4-225-236. 1970.
E Although Fenchel used a seawater medium
and inoculum, freshwater sources would
be equally useful.
F In these microaquaria simple ecosystems
develop when they are kept in complete
darkness. Complex photosynthetic
communities develop when they are
illuminated. A natural ecosystem is
figured by Fenchel (Figure 4), and a
related food web is shown in Figure 5
(both from Fenchel).
G Microaquaria Using Plastic Petri Dishes
Sessile ciliates have been successfully
collected, cultured, and used for bioassay
using the same petri dish (membrane filter
type, with tight fitting lids).
H Microaquaria using silicone cement rings
which allow diffusion of gases through the
silicone cultures will thereby remain
active indefinitely.
ADDITIONAL REFERENCES
1 Hutner, S.H. Protozoa as Toxicological
Tools. The Jour, of Protozoology.
11(1) 1-6. 1964.
2 Spoon, D.M. and Burbanck, W.D.
A New Method for Collecting Sessile
Ciliates m Plastic Petri Dishes with
Tight-Fitting Lids. Jour, of
Protozoology 14(4) 735-739. 1967.
5 Curds, C.R. and Cockburn, A.
II. Protozoa as Indicators in the
Activated Sludge Process. Water
Research 4 237-249. 1970.
6 Curds, C.R. An Illustrated Key to the
British Freshwater Ciliated Protozoa
Commonly Found in Activated Sludge.
Water Poll. Research Laboratory.
Stevenage, England. 90 pp.
i
7 Fenchel, Tom. The Ecology of Marine
Microbenthos. IV. Structure and
Function of the Benthic Ecosystem,
its Chemical and Physical Factors
and the Microfauna Communities with
Special Reference to the Ciliated
Protozoa. Ophelia 6 1-182. 1969.
8 Hutner, S. H. Botanical Gardens and
Horizons in Algal Research. In Challenge
for Survival. Pierre Dansereau ed.
Columbia University Press. 1970.
9 Hutner, S.H. ; Baker, H. , and Cox, D.
Nutrition and Metabolism in Protozoa.
Chapter in Biology of Nutrition, pp. 85-177.
edited by R.N. Fiennes. Pergamon Press.
1972.
10 Hutner, S.H. The Urban Botanical Garden-
An Academic Wildlife Preserve. Garden
Journ. 19(2):37-40. 1969.
This outline was prepared by Ralph M. Sinclair,
Aquatic Biologist, National Training Center,
Water Programs Operations, EPA, Cincinnati,
OH 45268.
-------
Bacteria and Protozoa as Toxieological Indicators
WfNOGRADSKy COLUMN
GENERALIZED
green
FILAMENTOUS GREEN ALGAE
green
SUPERNATE
COCCOID GREEN ALGAE
dull green
magenta
w hite
red
gyttja
4 GREEN PHOTO
INORGANIC SUBSTRATE
DIATOMS
BLUEGREEN ALGAE
NON-SULFUR PHOTO
FILAMENTOUS SULFUR
PURPLE SULFUR
BACTERIA
FIGURE 1
-------
Bacteria and Protozoa as Toxicological Indicators
1 cm
I i sea-water
agar with CaSQ^
glucose and
• peptone
0 5 >0 <5 days
FIGURE 2
A micro aquarium fitted with electrodes and the redox conditions through 17 days.
FIGURE 3
Drawing made by tracing a micrograph of the bacterial plate in a micro aquarium
(same experiment as shown on Figure 2.) Most conspicuous are the filaments of
Beggiatoa and the ciliates Cyclidium citrullus, Euplotes elegans and Holosticha
sp. Below the Oscillatoria filament (lower left) a Plagiopogon loricatus is seen.
Bacteria (except Beggiatoa) are not shown.
-------
Bacteria and Protozoa as Toxicological Indicators
FIGURE 4
The microflora and fauna in the surface of the Beggiatoa patches. (Oscillatoria,
Beggiatoa, Thiovolum, diatoms, euglenoids, nematode, Tracheloraphis sp.,
Frontonia marina, Diophrys scutum, Trochiloides recta).
METOPUS SPP
STROMBIDIUM SPP
ASPIDISCA
SEP
FUSCUM
PLAGIOPQGQN
LQRICATUS
-iEP
DISCOLOR
FIGURE 5
The food relationships of the most common ciliates in "estuarine" sediments and in sulphureta.
-------
ENVIRONMENTAL REQUIREMENTS OF FRESH-WATER INVERTEBRATES
L. A. Chambers, ^Chairman
Bacteria-Protozoa as Toxicological Indicators in Purifying Water
S. H. Hutner, Herman Baker, S. Aaronson and A. C. Zahalsky*
There is a cynical adage that all travelers
become entomologists. But, now with DDT
and detergents, travelers and stay-at-homes
alike are becoming toxicologists. We have
an immediate interest in pollution problems
Our laboratory receives, like the East River
and adjoining United Nations buildings, a
generous sootfall from a nearby power plant.
Also, we have seen a superb fishing ground,
Jamaica Bay in New York City, become a
sewer. (Jamaica Bay is, however, being
restored to its pristine cleanliness--but not
the U. N. area.) We take our theme
nevertheless not from aesthetics but from
statements by Berger (1961): (1) It is an
expensive, time-consuming project". . .to
predict with confidence a new waste's
probable impact on certain important down-
stream water uses. " And (2) "The
toxicological phase of the study is perhaps
its most perplexing aspect. The specialized
services and cost necessary for determining
the effect of repeated exposure to low con-
centrations of the waste for long periods of
time would inevitably place this job out of
reach of most public agencies. Equally
discouraging, perhaps, is the probability
that the toxicological study may take as long
as two years."
As described here, advances in microbiology
offer hopes of lightening this burden. The
first question is What kind of microcosm
will serve for toxicological surveys, especially
for predicting the poisoning of biological means
of waste disposal*5 The second is: Can the
protozoa of this microcosm predict toxicity
to higher animals'?
The food-chain pyramids of sewage plants
and polluted waters have been adequately
described (Hynes, I960, Hawkes, 1960).
A problem treated here is how to scale those
microcosms to experimentally manipulable
microcosms yielding dependable predictions
for the behavior of sewage-plant microcosms.
THE WINOGRADSKY COLUMN AS SOURCE
OF INOCULA FOR MINERALIZATIONS AND
AS TOXICOLOGICAL INDICATOR SYSTEM
Total toxicity depends on intrinsic toxicity-
persistence relationship. Techniques for
testing the degradability of organic compounds--
and so their persistence m soil and water- -
are still haphazard. The enrichment culture
technique, in which one seeks microbes that
use the compound in question as sole substrate--
hence degrades it and even "mineralizes"
it--was developed by the Dutch school of
microbiologists. Enrichment cultures are
used routinely by biochemists wishing to
work out the microbial catabolic metabolic
pathway for a compound of biochemical
interest. Since the compounds dealt with by
biochemists are of biological origin, micro-
bial degradability can be assumed. Still,
finding a microbe to degrade a rare biochemical
is not always easy Dubos, m a classical
hunt for a microbe able to live off the capsules
of pneumococci, found the bacterium only
after a long search which ended in a cranberry
bog. Such difficulty in finding microbes that
degrade rare biochemicals implies an even
greater difficulty in finding microbes that
degrade many products of the synthetic
^Director, Allan Hancock Foundation and Head, Bio. Dept., Univ. South. Calif.
+Haskins Laboratories, 305 E. 43rd St., New York 17, N.Y., and Seton Hall College
of Medicine and Dentistry, Jersey City, N. J. Pharmacological work from Haskins
Laboratories discussed here was assisted by grant R6-9103, Div. of General Medical
Sciences of the National Institutes of Health. Paper presented by S. H. Hutner.
-------
Environmental Requirements of Fresh-Water Invertebrates
organic chemicals industry, since such
compounds may embody biochemically rare
or biochemically nonexistent linkages.
Intimations of the importance of inoculum
abound in the literature, e.g., Ross and
Sheppard (1956) could not at first obtain
phenol oxidizers from ordinary inocula
(presumably soil and water), but manure
and a trickling filter from a chemical plant
proved abundant sources of active bacteria.
One wonders how extensive a study underlies
the statement quoted by Alexander (1961) that
"soils treated with 2,4, 5-T (trichlorophenoxy-
acetic acid) still retain the pesticide long
after all vestiges of toxicity due to equivalent
quantities of 2, 4-D have disappeared. "
What then is a reasonable inoculum for testing
a compound's susceptibility to microbial
attack? The size range is wide from the
traditional crumb or gram of soil or mud to
the scow-load of activated sludge contributed
by New York City to inaugurate the Yonkers
sewage-disposal plant. We suggest that to
strike a practical mean in getting a profile
of soil, mud, or sludge to be used as inoculum
the uses of the Winogradsky column should be
explored. Directions for Winogradsky column
and bacteriological enrichments have been
detailed (Hutner, 1962) and so only an outline
is given here. The column is prepared by
putting a paste of shredded paper, CaCO3>
and CaSO^ at the bottom of a hydrometer jar,
filling the jar with mud smelling H^S, covering
with a shallow layer of water, and illuminating
from the side with an incandescent lamp. In
2 or 3 weeks sharp zones appear- a green-
and-black anaerobic zone at the bottom
(a mixture of green photosynthetic bacteria
along with SO^-reducers, methane producers,
and the like), over that a red zone
(predominantly photosynthetic bacteria),
above this garnet or magenta spots or zone
(predominantly non-sulfur photosynthetic
bacteria), above this a layer rich in blue-
green algae (the transition to the aerobic
zones), above this aerobic bacteria along with
green algae, diatoms, other algae, and an
assortment of protozoa. This makes an
excellent simple classroom experiment to
demonstrate the different kinds of photo-
synthetic organisms, especially the bacterial
forms that are important in photosynthesis
research and that ordinarily escape notice
yet are ubiquitous in wet soils and natural
waters.
As pomted out by our colleague, Dr. L.
Provasoli (1961), the heterotrophic
capacities of algae are very imperfectly
known. This is underscored by recent
studies of the green flagellate Chlamydomonas
mundana as a dominant in sewage lagoons in
the Imperial Valley of California (Eppley and
MaciasR, 1962), other than that it prefers
acetate among the few substrates tried, its
heterotrophic capacities are unknown.
More unexpectedly, some strains of the
photosynthetic bacterium Rhodopseudomonas
palustris use benzoic acid anaerobically as
the reductant in photosynthesis (Scher, Scher,
and Hutner, 1962) narrowing the gap between
the photosynthetic pseudomonads and the
ubiquitous pseudomonads so often represented
among bacteria attacking resistant substrates
(e.g., hydrocarbons) as well as highly
vulnerable substrates. For the widely
studied, strongly heterotrophic photosynthetic
flagellates Euglena gracilis and E. viridis,
common in sewage, no specific enrichment
procedure is known, meaning that their
ecological niches are unknown but laboratory
data provide hints.
The increasing use of oxidation ponds would
in any case urge a greater use of scaled-
down ecological systems in which development
of none of the photosynthesizers present in
the original inoculum was suppressed.
Conceivably, some of the rare microbes
attacking rare substrates--and such microbes
are likely to represent a source of degraders
of resistant non-biochemicals--are specialists
in attacking products of photosynthetic
organisms.
Traditionally, the inoculum for a Winogradsky
column is a marine or brackish mud (as
New Yorkers we would be partial to mud from
flats of the Harlem River). Little is known
about the effectiveness as inocula of freshwater
muds or water-logged soils. It would be
valuable to know how complete a column
could develop from material from a trickling
filter or an activated-sludge plant. A practical
issue is- Might the poisoning of a sewage-
oxidation system be paralleled by the poisoning
of a Winogradsky column, where the poison
was mixed with the inoculum for the column9
Might the variously colored photosynthetic
zones of the column and the aerobic population
on top provide sensitive indicators for the
-------
Environmental Requirements of Fresh-Water Invertebrates
performance of a sewage-oxidation system
subjected to chemical wastes'5
If a particular compound mixed with inoculation
mud or sludge suppressed development of the
full Winogradsky pattern, one might assume
that the compound at the test concentration
was poisonous and persistent. Biological
destruction of such poisons, if at all possible,
might demand a long hunt for suitable micro-
organisms, then buildup of the culture to a
practical scale. This might best be done with
illuminated shake or aerated cultures, with
the inocula coming from a variety of environ-
ments. Optimism that microbes can be found
capable of breaking almost all the linkages of
organic chemistry is fostered by the study of
antibiotics, which include a wealth of
previously "unphysiological" linkages--azo
compounds, oximes, N-oxides, aliphatic and
aromatic nitro and halogen compounds, and
strange heterocyclic ring systems. Some
natural heterocycles, e.g., pulcherrimmic
acid and 2-n-nonyl-4-hydroxyqumolme, have
a disquieting resemblance to the potent
carcinogen 4-mtroquinoline N-oxide.
PROTOZOA AS TOXICOLOGICAL TOOLS
A difficult problem is one mentioned earlier,
persistence joined with low-grade toxicity to
higher animals. Recent developments in the
use of protozoa as pharmacological tools show
that protozoa can serve as sensitive detectors
of metabolic lesions ("side actions"9) of a
wide assortment of "safe" drugs. The list
includes the "anticholesterol" triparanol
(MER/29). Triparanol toxicity manifested
itself with several protozoa, including
Ochromonas danica (Aaronson et al., 1962)
and Tetrahymena (Holz et al., 1962).
Triparanol was not acting simply as an anti-
cholesterol for its obvious toxicity to protozoa
was annulled by fatty acids as well as by
sterols. The connection between the protozoan
results and the "side actions" of triparanol--
baldness, impotence, and cataracts--are of
course unclear, but protozoal toxicity might
serve as an initial warning that it might not
be as harmless as assumed from short-term
experiments with higher animals.
The anticonvulsant primidone provides a
clear indication of how protozoa can be used
to pinpoint the location of a metabolic lesion.
Primidone had been known to cause folic acid-
responsive anemias. It is therefore easy to
find that with joint use of a thymine-dependent
Escherichia coll and the flagellate Cnthidia
fasciculata reversal of growth inhibition by
folic acid and related compounds permitted
the charting of interferences with the inter-
connected folic acid, bioptenn, and DNA
function (Baker et al., 1962), which amply
accounted for the megaloblastic anemias,
Lactobacillus casei, a bacterium much used
in chemotherapeutic research, was unaffected
by the drug.
In another instance, where the mode of action
of the drug in higher animals was unknown,
growth inhibition property of the anticancer
compound 1-ammocyclopentane- 1-carboxylic
acid was reversed for Ochromonas danica
by L-alanine and glycine, as was the inhibition
property of l-amino-3-methyl-cyclohexane-
1-carboxylic acid by L-leucine (Aaronson
and Bensky, 1962).
Growth inhibition of Euglena by the potent
carcinogen 4-nitroquinoline N-oxide was
annulled by a combination of tryptophan,
tyrosine, nicotinic acid, phenylalanine,
uracil (Zahalsky et al., 1962) and, in more
recent experiments, the vitamin K relative
phthiocol. These N-oxides are of interest
because of recent work indicating that perhaps
the main way in which the body converts such
compounds as the amino hydrocarbons to the
actual carcinogens may be by an initial
hydroxylation of the nitrogen, e.g., work
by Miller et al., (1961). Whether the
peroxides in photo-chemical smogs of the
Los Angeles type act on hydrocarbons to
produce carcinogenic N-oxides is entirely
unknown. Leighton (1961) lists an array of
peroxy reactions produced by sunlight in
polluted air.
Our aforementioned experience with primidone,
a ketonic heterocycle, led us to test the
sedative thalidomide. It was toxic for
Ochromonas danica 0_. malhemensis and
Tetrahymena pyriformis, this toxicity was
-------
Environmental Requirements of Fresh-Water Invertebrates
annulled by nicotinic acid (or nicotinamide)
or vitamin K (menadione) (Frank et al, 1963).
We do not know whether a similar protection
could have been conferred on human embryos
or polyneuritis in the adult.
Many widely used herbicides of the dinitrophenol
type are powerful poisons for higher animals.
We do not know how sensitive protozoa would
be for detecting their persistence. However,
since somewhat similar thyro-active com-
pounds can be sensitively detected by their
exaggeration of the B^ requirement of
Ochromonas malhamensis (Baker et al, 1961),
this flagellate might be a useful test object
for dinitrophenols and congeners.
The Paramecium (and perhaps too the
Tetrahymena) test for polynuclear benzenoid
carcinogens has remarkable sensitivity and
specificity (Epstein and Burroughs, 1962, Hull,
1962). This test depends on the carcinogen-
sensitized photodynamic destruction of
paramecia by ultraviolet light. This test is
approaching practicality for air, and there is
no reason to suppose it cannot be applied to
benzene extracts of foodstuffs and water.
CONCLUSIONS
We have suggested here new procedures for
examining the intrinsic toxicity-persistence
relationship, using as test organisms the
protists represented conspicuously in a
Winogradsky column. The new field of
micro-toxicology is virtually undeveloped.
The urgent need for detecting chronic, low-
grade toxicities is evident from many sides.
This is not the place for a detailed discussion
of the medical implications of this area of
research, but it should be emphasized that
chronic toxicities and carcinogenesis are
related. Conversely, Umezawa (1961) has
remarked that most antitumor substances
have chronic toxicities and that elaborate
testing procedures for toxicity are required
to fix the daily tolerable dose, apparently this
problem is a central theme in medical as well
as pollution research. Inhibition of growth of
an array of protozoa is now in practical use
as a means of detecting anticancer substances
in antibiotic beers (Johnson et al, 1962).
Since the embryos appear to lack the
detoxication mechanisms of the adult animal
(Brodie, 1962), toxicity for protozoa (which
presumably lack these detoxication
mechanisms) should be compared with that
for the embryo, not the adult, as emphasized
by the thalidomide disaster.
There are further limitations on the use of
microbes as detectors of toxicity. High-
molecular toxins seem inert to microbes,
and antihormones (with the exception of
anti-thyroid compounds) are generally inert.
The main usefulness of microbial indices of
toxicity would appear, then, to be for detecting
low-molecular poisons acting on cellular
targets rather than on cell systems and
organs. These are precisely the poisons
likely to put out of business a pollution-
control installation primarily dependent on
microbial activity.
REFERENCES
Aaronson, S. and Bensky, B. 1962.
Protozoological studies of the cellular
action of drugs. I. Effect of
1-aminocyclopentane- 1-carboxylic acid
and l-amino-3-methylcyclo-hexanel-1 -
carboxylic acid on the phytoflagellate
Ochromonas danica. Biochem. Pharmacol.
11:983-6.
Aaronson, S., Bensky, B., Shifrine, M. and
Baker, H. 1962. Effect of hypocholes-
teremic agents on protozoa. Proc. Soc.
Exptl. Biol. Med. 109-130-2.
Alexander, M. 1961. "Introduction to Soil
Microbiology", John Wiley &. Sons, N. Y.
(see p. 240).
Baker, H., Frank, O., Hutner, S. H.,
Aaronson, S., Ziffer, H. and Sobotka, H.
1962. Lesions in folic acid metabolism
induced by primidone. Experientia
18.224-6.
Baker, H., Frank, O., Pasher, I., Ziffer, H.,
Hutner, S.H. and Sobotka, H. 1961.
Growth inhibition of microorganisms by
thyroid hormones. Proc. Soc. Exptl.
Bio. Med. 107 965-8.
-------
Environmental Requirements of Fresh-Water Invertebrates
Berger, B.B. 1961. Research needs in water
quality conservation. In "Algae and
Metropolitan Wastes", Trans. 1960
Seminar, Robt. A. Taft Sanitary Eng.
Center, Cincinnati 26, Ohio, p. 156-9.
Brodie, B. D. 1962. Drug metabolism-
subcellular mechanisms. In "Enzymes
and Drug Action", J. L. Mongar and
A.V. S. de Reuck, eds., Ciba Foundation
Symposium, J. & A. Churchill Ltd.,
p. 317-40.
Eppley, R.W. and MaciasR, F.M. 1962.
Rapid growth of sewage lagoon
Chlamydomonas with acetate. Physiol.
Plantarum 15-72 - 9.
Epstein, S. S. and Burroughs, M. 1962.
Some factors influencing the photodynamic
response of Paramecium caudatum to
3,4-benzypyrene. Nature 193:337-8.
Frank, O., Baker, H., Ziffer, H.,
Aaronson, S., and Hutner, S.H. 1963.
Metabolic deficiencies m protozoa induced
by Thalidomide. Science 139 110-1.
Hawkes, H.A. 1960. Ecology of activated
sludge and bacteria beds. In "Waste
Treatment", P,C.G. Isaac, ed.,
Pergamon, N.Y., Oxford, etc., p. 52-97,
discussion p. 97-8.
Holz, G.G., Jr., Erwin, J., Rosenblum, N.
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