PLANKTON ANALYSIS
TRAINING MANUAL
FEDERAL WATER POLLUTION CONTROL ADMINISTRATION
U.S. DEPARTMENT OF THE INTERIOR
-------
PLANKTON ANALYSIS
This course is offered for professional personnel in
the fields of water pollution control, limnology, and
also water supply. Primary emphasis is given to
practice in the identification and enumeration of or-
ganisms which may be observed in the microscopic
examination of water. Problems of significance and
control are also considered.
U.S. DEPARTMENT OF THE INTERIOR
Federal Water Pollution Control Administration
TRAINING PROGRAM
May 1970
-------
FOREWORD
These manuals are prepared for reference use of students enrolled
in scheduled training courses of the Federal Water Pollution Control
Administration.
Due. to the. limited pn.odu.at4.on and availability
at the. ma.nua.tA, it it not appiopiiate. to c-Lte.
the.m at tic.hnic.al teje'ienceA in bibliogiaphie.4
hi othe.fi fioimt oj publication.
Re.6e.ie.nc.e.t to pn.odvic.ti> and manu6ac.tuie.it it
illuttiation onty; tuck ie.$e.ie.nc.e.t do not imply
product e.ndon.te.me.nt by the. fe.de.iai Wate.1 Pollution
Control tidmini&tiation on. the. U.S. Ve.paitme.nt
0(5 Ake. lnte.iioi.
The reference outlines in this manual have been selected and developed
with a goal of providing the student with a fund of the best available
current information pertinent to the subject matter of the course. Indi-
vidual instructors may provide additional material to cover special
aspects of their own presentations.
This manual will be useful to anyone who has need for information on the
subjects covered. However, it should be understood that the manual will
have its greatest value as an adjunct to classroom presentations. The
inherent advantages of classroom presentation is in the give-and-take
discussions and exchange of information between and among students and
the instructional staff.
Constructive suggestions for improvement in the coverage, content, and
format of the manual are solicited and will be given full consideration.
H. M. Freeman
Chief, Direct Training Branch
Division of Manpower and Training
Federal Water Pollution Control
A dministration
-------
TRAINING PROGRAM
The Federal Water Pollution Control Administration of the U. S. Department
of the Interior conducts programs of research, technical assistance, enforce-
ment, and technical training for water pollution control.
Training is available at five installations of the Administration. These are:
the National Training Center located at the Robert A. Taft Sanitary Engineering
Center in Cincinnati, Ohio; the Robert S. Kerr Water Research Center, Ada,
Oklahoma; the Southeast Water Laboratory, Athens. Georgia; the Pacific
Northwest Water Laboratory, Corvallis, Oregon, and the Hudson-Delaware
Basins Office, Edison, New Jersey.
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. These courses are being
designed for nontechnical audiences, including administrators at the policy and
decision-making levels, representatives of public action groups, and others not
requiring the depth of detail of the more specialized courses.
Scientists, engineers, and recognized authorities from other FWPCA 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 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 in
the planning and development of operator-training courses. Guest appearances
of instructors from the Federal Water Pollution Control Administration, and the
loan of instructional materials such as lesson plans and visual training aids, may
be obtained through special arrangement. These training aids, including
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.
An annual Bulletin of Courses is prepared and distributed by the Water Pollution
Control Training Program. 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 one
of the training centers mentioned above.
-------
D£PAKTMENT OF THE
MANPOWER AND TRAINING DIVISION
DIRECT TRAINING BRANCH
H. M. Freeman, Chief
TRAINING ACTIVITIES OF THE ADMINISTRATION
SOUTHEAST WATER LABORATORY
Athens, Georgia
R. Roth, Sanitary Engineer, Chief
W. R. Davis, Chemist
ROBERT S. KERR WATER RESEARCH CENTER
Ada, Oklahoma
(Mrs.)M. E. Smith, Sanitary Engineer, Chief
J. E. Matthews, Aquatic Biologist
ROBERT A. TAFT SANITARY ENGINEERING CENTER
Cincinnati, Ohio
H. L. Jeter, Microbiologist, Director
(Miss) A. E. Donahue, Chemist
C R. Feldmann, Chemist
P. F. Hallbach. Chemist
H. W. Jackson, Chief Biologist
F. J. Ludzack, Chemist
R. Russomanno, Microbiologist
R. M. Sinclair, Aquatic Biologist
C. E Sponagle. Sanitary Engineer
PACIFIC NORTHWEST WATER LABORATORY
Corvallis, Oregon
L. J. Nielson, Sanitary Engineer, Chief
D. S. May, Microbiologist
J. Wooley, Aquatic Biologist
HUDSON-DELAWARE BASINS OFFICE
Edison, New Jersey
F. P. Nixon, Deputy Regional Training Officer
R. B. Fagan, Aquatic Biologist
10.69
-------
PLANKTON ANALYSIS (141)
2 weeks
CINCINNATI, OHIO May 11-22, 1970
October 5-16,1970
This course is offered for professional personnel
concerned with the evaluation of natural and polluted
waters by means of plankton examination. Limited
attention is also devoted to the examination and inter-
pretation of the fauna of activated sludge and waste
stabilization ponds.
Instruction enables the student to carry out basic
laboratory procedures in the identification and counting of
both phytoplankton and zooplankton. He will be capable
of applying laxonomic proceduresto plankton and recognize
the major types he is likely to encounter. He will be
able to calibrate a microscope and to carry counting and
group identification to the point of obtaining results
which are qualitatively and quantitatively reliable.
Attention is gixen to the significance of various
types of counts. Forms frequently found in water and
wastewater treatment plants and polluted environments
are emphasized. Techniques for plankton control are
presented. Time is provided for discussion of local
problems, both in class and with specialists at the train-
ing facility.
Representative course topics usually include
Water quality problems of biological origin
Identification of planktonic animals and plants
(a series of lectures and laboratories com-
prising approximately half of the course)
Microscope calibration
Plankton analysis
Sampling and preparation
Techniques of counting
(enumeration, methods selection
Plant operation problems
Plankton in stabilization ponds
Activated sludge fauna
Toxic algae
Other biological treatment problems
Plankton control
Plant control
Control in surface waters
Although microscopes arc available for class use,
more effective training results when it is given on the
same instrument that will be used in the home laboratory.
The microscope should have magnifications up to approx-
imately 400X, oil immersion is optional. The student
consequently is urged to hand-carry his own microscope
to the course.
-------
CONTENTS
Title or Description Outline Number
CHAPTER I INTRODUCTION
Water Resources and Needs 1
Limnology and Ecology of Plankton 2
Optics and the Microscope 3
The Aquatic Environment 4
CHAPTER H IDENTIFICATION OF PLANKTON AND
ASSOCIATED ORGANISMS
Structure and Function of Cells 1
Aquatic Organisms of Significance in Plankton Surveys 2
Types of Algae 3
Blue-Green Algae 4
Green and Other Pigmented Flagellates 5
Filamentous Green Algae 6
Coccoid Green Algae 7
Diatoms 8
Filamentous Bacteria 9
Protozoa, Nematodes, and Rotifers 10
Free-Living Amoebae and Nematodes 11
Animal Plankton 12
Laboratory Exercises
General Laboratory Instructions 15
Types of Algae 16
Identification of Diatoms 17
Identification of Animal Plankton 18
141.5.70
-------
Contents
Title or Description Outline Number
CHAPTER HI TECHNIQUES OF PLANKTON METHODOLOGY
Techniques of Plankton Sampling Programs 1
Preparation and Enumeration of Plankton in the Laboratory 2
Calibration and Use of Plankton Counting Equipment 3
Preparation of Permanent Diatom Mounts 4
Determination of Odors 5
Collection and Interpretation of Biological Lake Data 6
Determination of Plankton Productivity 8
Methods of Measuring Standing Crops of Plankton 9
Aerial Reconnaissance in Pollution Surveillance 10
Laboratory Exercises
Proportional Counting of Plankton 11
Calibration of Plankton Counting Equipment 12
Fundamentals of Quantitative Counting 13
Class Problem in Plankton Analysis 14
CHAPTER IV INTERPRETATION AND SIGNIFICANCE OF PLANKTON
Algae and Actinomycetes in Water Supplies 1
Algae as Indicators of Pollution 2
Public Health Significance of Toxic Algae 3
Odor Production by Algae and Other Organisms 4
Organic Enrichment and Dissolved Oxygen Relationships in Water 5
Plankton in Oligotrophic Lakes 6
The Effects of Pollution on Lakes 7
-------
Contents
Title or Description Outline Number
CHAPTER V PLANKTON CONTROL
Control of Plankton in Surface Waters 3
Control of Interference Organisms in Water Supplies 4
Nutrients: The Basis of Productivity 5
CHAPTER VI RELATED STUDIES
The Problem of Synthetic Organic Wastes 2
Beneficial Aspects of Algae 3
Behavior of Radionuclides in Food Chains - Freshwater Studies 4
FWPCA Responsibilities for Water Quality Standards 5
Marine and Estuarine Plankton 6
Attached Growths (Periphyton or Aufwuchs) 7
Artificial and Related Substances ~ References 8
CHAPTER VII IDENTIFICATION KEYS
Key to Selected Groups of Freshwater Animals 1
Key to Algae of Importance in Water Pollution 2
APPENDIX
Foreword
Classification-Finder for Names of Aquatic Organisms
in Water Supplies and Polluted Waters
-------
Intervale
Reservoir
CHAPTER I
INTRODUCTION
Water Resources and Needs
Limnology and Ecology of Plankton
Optics and the Microscope
The Aquatic Environment
1
2
3
4
-------
WATER RESOURCES AND NEEDS
I WATER RESOURCES
A The source of all freshwater is the
hydrologic cycle, shown in Figure 1.
CIRCULATION
EVAPORATION
\ \ -= -^ I WATER TABLE
\ IMPOUNDMENT
GROUNDWATER GROUNDWATER
THE HYDROLOGIC CYCLE
Figure 1
1 Precipitation of water as rain, snow,
hail, sleet or dew.
2 Percolation of water through soil to an
aquifer to form groundwater.
3 Runoff of water which forms lakes,
streams and rivers.
4 Evaporation of surface water or trans-
piration of water from green plants to
the atmosphere.
5 Atmospheric recirculation of the water
vapor.
B The world's supply of water is contained
within the hydrologic cycle as:
1 Oceanic water
2 Water vapor in the atmosphere
3 Ice and snow in glaciers and snowpack
4 Runoff water in lakes and streams
5 Groundwater
C Withdrawals for use are mostly from those
waters in the runoff and groundwater
phases, although some oceanic waters are
being utilized.
D Precipitation--which serves to recharge
groundwaters and surface supplies--is at
a relatively fixed annual rate.
1 Average precipitation in the U.S. is
30 inches per year or 3, 900 billion
gallons per day.
2 Evapo-transpiration losses total
approximately 21 inches per year or
approximately 2, 740 billion gallons per
day.
3 The available water totals approximately
9 inches per year or 1, 160 billion
gallons per day.
II THE DISTRIBUTION OF U.S. WATER
RESOURCES
Although the water supply in the hydrologic
cycle is fixed in amount, it is not distributed
evenly. A wide disparity of water distribution
exists both in time and space. Distribution
of the annual average precipitation is shown
in Figure 2.
A Distribution of Precipitation
1 Dependent upon:
a Atmospheric conditions such as
temperature and winds
b The geography of the region
c The general climate of the area
2 U. S. areas of high annual precipitation
a The Pacific slope varies from 10 inches
to greater than 100 inches annually.
W.RE.28d.4.70
I 1-1
-------
Water Resources and Needs
Distribution of Precipitation
(Average Annual)
Inciies
Figure 2
b The gulf states precipitation varies
from 20 to 60 inches annually.
c Precipitation in the midwest and
Great Lakes area ranges from 25 to
50 inches per year.
d Precipitation along the Atlantic Coast
averages between 35 to 50 inches
per year.
3 Areas of low annual precipitation
a The Rocky Mountain area precipitation
ranges between 10 and 20 inches per
year.
b Much of the southwest has less than
10 inches of precipitation annually.
4 Distribution of precipitation with time
a The rainy or wet season varies from
summer to winter, or in some areas
there is relatively little change
throughout the year.
b Local storms of high intensity may
reach as much as 30 inches in 24
hours.
B Distribution of Runoff
1 Dependent upon:
a Precipitation in the region
b Infiltration - which is controlled by
the geologic formations and the time
lapse between rains.
c Season of the year controls evaporation,
and snow melt.
d Topography controls the time available
to percolate through the soil.
e Vegetation type and density affects
interception and evapotranspiration.
2 Areas of high annual runoff
a Sections of the Pacific slope have
greater than 80 inches annually.
-------
Water Resources and Needs
b The eastern 1/3 of the U.S. averages
greater than 20 inches of runoff
annually.
3 Much of the western U.S.has less than
1 inch of runoff annually.
a Southwest
b Rocky Mountain states
c Rocky Mountain plateau
4 Time distribution of runoff
a Overflow--runoff during and immediately
following precipitation.
b Base flow--sustained or fair weather
runoff composed of delayed sub-
surface and groundwater runoff.
See Figure 3 for runoff cycle.
C Distribution of Groundwater
1 Groundwater volume is affected by the
same factors as runoff.
2 Geologic formations and soils control
percolation and storage of groundwater.
3 Topography controls time available
for percolation.
4 Evapo-transpiration varies with the
season, as does precipitation and
ground saturation.
Ill WATER USE
A Present Water Use in the U. S.
1 Water available for use
a Nine inches or 1, 160 billion gallons
per day are not lost through evapo-
transpiration, and is therefore
theoretically available.
b Water use in the U.S. at the present
time is approximately 390 billion
gallons per day or 3 inches of our
total supply.
c Twenty-one inches are lost through
evapo-transpiration.
storage i Surface detention = sheet of water
lnfiltratipn!J?r"*8^S&»4^La»J£ Overland flow
Surface runoff
Perchea""wate'r table ^1 mpervfou's
^_ SS^K.
Water table -===
Ground-water flow ^"^Stream channel
THE RUNOFF CYCLE
(Davis & DeWiest)
Figure 3
-------
Water Resources and Needs
2 The way in which water is used
Water uses can be grouped into two
classes. Those uses which are in situ
such as recreation, fishing, and wildlife
and those uses requiring withdrawal
from the stream. These withdrawals are:
a Agricultural uses take 46% of our
supply or 180 billion gallons/day;
only 40% of this water is returned to
the streams.
b Industrial uses take another 46% of our
supply. 2% of the water used by
industry is consumed.
c Municipal uses total approximately
25 billion gallons daily or 8% of the total.
3 Source of water used in U. S.
a National averages show 80% or 312
billion gallons per day to be from
surface sources, while 20% is taken
from the ground.
b The ratio of surface water to ground-
water varies and is dependent on the
quantity and quality available in each
locality, as well as the cost.
4 Seasonal uses of water
a Irrigation waters are used during the
growing season only.
b Some water using industries such as
the canning industry are seasonal.
c The majority of industries needs
water throughout the year.
d Municipal use is higher in the summer.
B Demand for water is increasing
1 The predicted demand of water in 1980
is approximately 600 billion gallons of
water per day, or 220, 000 billion per
year.
2 This is mainly due to expansion of
industry and irrigated agriculture.
3 Much of the demand for water will be
in areas such as the southwest, that
are already short on water.
C Methods for the Development of U. S.
Water Resources for Future Needs
1 Utilization of our present sources of
water, surface and groundwater, must
be increased. This would mean
increased storage, both on the surface
and in underground reservoirs.
2 Desalinization of ocean waters and
brackish waters holds some promise
for regions where transportation will
not be expensive.
3 Reduction of evapo-transpiration losses
will greatly increase our total available
supply.
4 Weather modification methods could
possibly give us precipitation in the
right place at the right time.
5 Greater reuse of our present supply is
both through multiple use and better
waste treatment methods.
IV SUMMARY
The total amount of water available appears
to be fixed. In view of the increasing
demands and the currently inefficient
utilization of the supply, the demand may
very shortly exceed the supply. Better
management of the resource and more
engineering research are urgently needed.
I 1-4
-------
Water resources and Needs
Table 1. AVAILABILITY OF GROUND WATER
Areas
A
B
C
D
£
F-l
F-2
G
Atlantic and Gulf Coastal Plain area
Southern Great Plains area
Appalachian Mountain and Piedmont area
Rocky Mountains, northern Great Plains,
and northern Pacific Coast area
Unglaciated central plateaus and lowlands
Basin and range
Columbia Plateau
Glaciated area of the East and Midwest
U. S. Total (rounded)
Water Use
(excluding water power)
Use in mgd and Percent
of total from Ground
Water Sources
Total
mgd
32,000
21.000
8,000
28, 000
26,000
41, 000
24, 000
57, 000
240, 000
Ground
water (%)
25
45
50
12
10
42
7
10
20
A CKNOWLEDGEMENT:
Certain portions of this outline contains
training material from prior outlines by
Peter F. Atkins, F.P. Nixon.
REFERENCES
1 Ackerman, Edward A., Lof, George O.G.,
Technology in American Water
Development. The Johns Hopkins
Press. Baltimore. 1959.
2 Senate Select Committee on National Water
Resources: Water Resources Activities
in the United States: Committee Print
No. 3. U.S. Gov. Printing Office.
January 1960.
3 Senate Select Committee on National Water
Resources: Water Resources Activities
in the United States: Committee Print
No. 24. U.S. Gov. Printing Office.
January 1960.
4 Linsley, RayK.. Kohler. Max A.,
Paulttus, Joseph H. Hydrology for
Engineers. McGraw-Hill Book Co.,
Inc., New York. 1958.
5 Chow, Ven Te. Handbook of Applied
Hydrology. McGraw-Hill Book Co.,
Inc., New York. 1964.
6 Davis, Stanley N. and DeWiest, Roger,
J.M. Hydrogeology. John Wiley
and Sons, Inc., New York. 1966.
7 American Chemical Society. Cleaning
Our Environment the Chemical Basis
for Action. ACS. Washington, DC
20036. 249pp. (2.75) 1969.
This outline was prepared by Edward D.
Schroeder, Former Engineer, FWPCA
Training Activities, SEC and revised by
L. J. Nielson, Chief Technical Training,
Pacific Northwest Water Laboratory,
Corvallis, Oregon.
I 1-5
-------
LIMNOLOGY AND ECOLOGY OF PLANKTON
I INTRODUCTION
A Most Interference Organisms are
Small.
B Small Organisms generally have
Short Life Histories.
C Populations of Organisms with
Short Life Histories may Fluctuate
Rapidly in Response to Key Environ-
mental Changes.
D Small Organisms are Relatively
at the Mercy of the Elements
E The Following Discussion will
Analyze the Nature of These Ele-
ments with Reference to the Res-
ponse of Important Organisms.
II PHYSICAL FACTORS OF THE ENVIRON-
MENT
A Light is a Fundamental Source of
Energy for Life and Heat.
1 Insolation is affected by geo-
graphical location and mete-
orological factors.
2 Light penetration in water is
affected by angle of incidence
(geographical), turbidity, and
color. 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 arithmetically,
the light tends to decrease geo-
metrically. Blues, greens, and
yellows tend to penetrate most
deeply while ultra violet, vio-
lets, 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.
3 Turbidity may originate within
or outside of a lake.
a That which comes in from
outside (allochthonous) is
predominately inert solids
(tripton).
b That of internal origin (auto-
chthonous) tends to be bio-
logical m nature.
B Heat and Temperature Phenomena
are Important in Aquatic Ecology.
1 The total quantity of heat avail-
able to a body of water per year
can be calculated and is known
as the heat budget.
2 Heat is derived directly from in-
solation; also by transfer from
air, internal friction, and other
sources.
C Density Phenomena
1 Density and viscosity affect the
floatation and locomotion of
microorganisms.
Pure fresh water achieves
its maximum density at 4 C
and its maximum viscosity
at 0°C.
b The rate of change of density
increases with the temperature.
Density stratification affects
aquatic life and water uses.
a In summer, a mass of warm
surface water, the epilimmon.
is usually present and separated
from a cool deeper mass, the
hypolimmon, by a relatively
thin layer known as the
thermoclme.
b Ice cover and annual spring
and fall overturns are due to
successive seasonal changes
in the relative densities of
the epilimmon and the hypo-
BI. MIC.eco.4b.4.70
I 2-1
-------
Limnology and Ecology of Plankton
E
limnion, profoundly influ-
enced by prevailing meteoro-
logical conditions.
c The sudden exchange of
water masses having differ-
ent chemical characteris-
tics may have catastrophic
effects on certain biota.
d Silt laden waters may seek
certain levels, depending
on their own specific gravity
in relation to existing layers
already present.
e Saline waters will also
stratify according to the
relative densities of the
various layers.
3 The viscosity of water is greater
at lower temperatures.
a This is important not only
in situations involving the
control of flowing water as in
a sand filter, but also since
overcoming resistance to
flow generates heat, it is
significant in the heating
of water by internal friction
from wave and current ac-
tion and many delay the
establishment of anchor
ice under critical conditions.
b It is easier for plankton
to remain suspended in cold
viscous (and also dense)
water than in less viscous
warm water. This is re-
flected in differences in the
appearance of winter vs
summer forms of life (also
arctic vs tropical).
Shore development, depth, inflow -
outflow pattern, and topographic
features affect the behavior of the
water.
Water movements that may affect
organisms include waves, currents,
tides, seiches, and floods.
1 Waves or rhythmic movement
a The best known are traveling
waves. These are effective
only against objects near
the surface. They have little
effect on the movement of
large masses of water.
b Standing waves or seiches
occur in all lakes but are
seldom large enough to be
observed. An "internal seich"
is an oscillation in a density
mass within a lake with no
surface manifestation may
cause considerable water
movement.
2 Currents
a Currents are arhythmic
water movements which have
had major study only in ocean-
ography. They primarily are
concerned with the translo-
cation of water masses. They
may be generated internally
by virtue of density changes,
or externally by wind or
runoff.
b Turbulence phenomena or
eddy currents are largely re-
sponsible for lateral mixing
in a current. These are of
far more importance in the
economy of a body of water
tnan mere laminar flow.
F Surface Tension and the Surface Film
1 The surface film is the habitat
of the "neuston", a group of
particular importance in water
supplies.
2 The biological effects of small
amounts of detergents and simi-
lar agents are yet to be evaluated.
Ill DISSOLVED SUBSTANCES
A Carbon dioxide is released by plants
I 2-2
-------
Limnology and Ecology of Plankton
and animals in respiration, but
taken in by plants in photosynthesis.
B Oxygen is the biological complement
of carbon dioxide, and necessary
for all animal life.
C Nitrogen and phosphorus are fun-
damental nutrients for plant life.
1 Occur in great dilution, con-
centrated by plants.
2 The distribution of nitrogen com-
pounds is generally correlated
with the oxygen curve.
D Iron, manganese, sulphur, and
silicon are other minerals impor-
tant to aquatic life which exhibit
biological stratification.
E Many other minerals are present
but their biological distribution in
waters is less well known.
F Dissolved organic matter is present
in even the purest of lakes.
BIOLOGICAL FACTORS
A Nutritional Classification of Or-
ganisms
1 Holophytic or independent or-
ganisms, like green plants, pro-
duce their own basic food ele-
ments from the physical environ-
ment.
2 Holozoic or dependent organisms,
like animals, ingest and digest
solid food particles of organic
origin.
3 Saprophytic or carrion eating
organisms, like many fungi and
bacteria, digest and assimilate
the dead bodies of other organ-
isms or their products.
B The Prey-Predator Relationship
is Simply one Organism Eating
Another.
C Toxic and Hormomc Relationships
1 Some organisms such as certain
blue green algae and some ar-
mored flagellates produce sub-
stances poisonous to others.
2 Antibiotic action in nature is
not well understood but has been
shown to play a very influential
role in the economy of nature.
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.
1 Those that pass through a plankton
net (No. 25 silk bolting cloth or
equivalent) or sand filter are
known as nannoplankton (which
usually greatly exceed the net
plankton in quantity).
2 Those less than four microns
in length are sometimes called
ultraplankton.
3 There are many ways in which
plankton may be classified: taxo-
nomic, ecological, industrial.
4 The concentration of plankton varies
markedly in space and time.
a Depth, light, currents, and
water quality profoundly affect
plankton distribution.
b The relative abundance of
plankton in the various sea-
sons is generally:
1 spring, 2 fall, 3 summer,
4 winter
B Benthic organisms (benthos) are those
living on or near the bottom, frequently
attached. Typically benthic organisms
such as certain filamentous algae, may
on occasion be broken or washed free,
I 2-3
-------
Limnology and Ecology of Plankton
and mingled with the typical plankton.
C The emergent vascular shore plants
are often a very influential commun-
ity; on death and decay, they add
nutrients to the water.
D The shallow water or littoral com-
munity is one of the most varied and
productive areas - a fluctuating water
level will discourage this community.
E The deep water and sludge or ooze
communities may contribute tastes,
odors, and undesirable chemical
characteristics.
VI THE EVOLUTION OF WATERS
A The history of a body of water de -
termines its present condition.
Natural waters have evolved in the
course of geologic time to what we
know today.
B 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
streambed, 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 in-
crease in flow.
4 In old age, streams have approa-
ched base level. During flood
stage they scour their bed and de-
posit 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 tem-
porarily interrupted. Thus as essen-
tially "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 rever-
sion to the "original" condition.)
Geological factors which significantly
affect the nature of 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 pre-
cipitation.
6 The natural vegetative cover of
the land is of course responsive
to many of the above factors and
is also severely subject to the
whims of civilization. This is
one of the major factors deter-
mining run-off versus soil absorp-
tion, etc.
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.
I 2-4
-------
Limnology and Ecology of Plankton
2 Maturing 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 irreg-
ulars areas.
c Silt brought in by tributary
streams settles out in the
quiet lake water.
d Rooted aquatics 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 pre-
cipitated in the deeper
portions of the lake in part
through the action of plants.
f When filling is well advanced
sphagnum mats extend out-
ward 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 out-
let.
b Filling with detritus eroded
from the shores or brought
in by tributary streams.
c Filling by the accumulation of
the remains of vegetable
materials growing in the
lake itself.
(Often two or three pro-
cesses may act concurrently)
VI PRODUCTIVITY
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 productivity is a "poor"
water biologically, and also a rela-
tively "pure" or "clean" water; hence
desirable as a water supply. A
productive water on the other hand
may be a nuisance to man or highly
desirable. Some of the factors
which influence the productivity of
waters are as follows:
B Factors affecting stream productivity.
To be productive of plankton, a stream
must provide adequate nutrients, light,
a suitable temperature, and time for
growth to take place.
1 Youthful streams, especially
on rock or sand substrates are low
in essential nutrients. Tempera-
tures in mountainous 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, temperature 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 avail-
able for plankton growth increases,
the balance between turbidity,
nutrient levels, and temperature
and other seasonal conditions,
I 2-5
-------
Limnology and Ecology of Plankton
determines the overall produc-
tivity.
C Factors Affecting the Productivity
of Lakes
1 The size, shape, and depth
of the lake basin. Shallow
water is more productive than
deeper water since more light
will reach the bottom to stim-
ulate rooted plant growth. As
a 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 influenced 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.
3 Turbidity reduces productivity
as light penetration is reduced.
4 The presence or absence of
thermal stratification with its
semi-annual turnovers affect
productivity by distributing
nutrients throughout the water
mass.
5 Climate, temperature, pre-
valance 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
deliberately increase or decrease
productivity. This can be dem-
onstrated by a comparison of the
TVA reservoirs which practice a
summer drawdown with some of
those in the west where a winter
drawdo'vn is the rule.
The level at which water is re-
moved from a reservoir is also
important. The hypolimnion may
may be anaerobic while the epi-
limnion is aerobic.
Reservoir discharges also pro-
foundly affect the DO, temperature,
and turbidity in the stream below
a dam. Too much fluctuation in
flow may permit sections of the
stream to dry, or provide inade-
quate dilution for toxic waste.
VIII CLASSIFICATION OF LAKES AND RESER-
VOIRS
A The productivity of lakes and impound-
ments is such a conspicuous feature
that it is often used as a convenient
menas 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 - bog lakes -
low in pH, water yellow to brown,
dissolved solids, N, P, and Ca
scanty but humic materials abun-
dant, bottom fauna and plankton
poor, and fish species are limited.
B Reservoirs may be classified into
I 2-6
-------
FACTORS AFFECTING PRODUCTIVITY
Geographic Location
Human
Influence
Sewage
Agriculture
Mining
Primary
Nutritive
Materials
Latitude
Longitude
Altitude
Geological
Formation
Topography
Composition
of Substrate
Shape of Basin
Wind
Precipitation // I Insolation
Area Bottom
Conformation
Drainage
Area
Nature at
Bottom
Depo
S
Inflow of
Allochthonous
"Materials
. Trans-
parency
-Light
Penetration
leat Penetration
and Stratification-
i Penetra Develop
^- and Littoral
Utilization Region
Seasonal Cycle
Circulat. Stagnation
Growing Season
Trophic Nature of a Lake
to
I
-o
-------
Limnology and Ecology of Plankton
IX
two types, storage and run of the
river.
1 Storage reservoirs have a large
volume in relation to their in-
flow.
2 Run of the river reservoirs have
a large flow through in relation
to their storage value.
According to location, lakes and
reservoirs may be classified as
polar, temperate, or tropical.
Differences in climatic and geo-
graphic conditions result in dif-
ferences in their biology.
THE MANAGEMENT OR CONTROL OF
ENVIRONMENTAL FACTORS
A Liebig's Law of the Minimum states
that productivity is limited by the
nutrient present in the least amoung
at any given time relative to the
assimilative capacity of the organism.
B Shelford's Law of Toleration:
Minimum Limit
of toleration
Absent
Decreasing
Abundance
Range of Optimum
of factor
Greatest abundance
Maximum limit of '
toleration
Decreasing
Abundance
Absent
C The artificial introduction of nutrients (sewage pollution or
C The artificial introduction of
nutrients (sewage pollution or fer-
tilizer) thus tneds to eliminate ex-
isting limiting mmimums for some
species and create intolerable maxi-
mums for other species.
1 Known limiting mmimums may
sometimes be deliberately
maintained.
2 As the total available energy
supply is increased, productivity
tends to increase.
3 As productivity increases, the
whole character of the water
may be changed from a meagerly
productive clear water lake
(oligotrophic) to a highly pro-
ductive and usually turbid lake
(eutrophic).
4 Eutrophic ation leads to treatment
troubles.
D Control of eutrophication may be
accomplished by various means
1 Watershed management, ade-
quate preparation of reservoir
sites, and pollution control tend to
maintain minimum limiting nu-
tritional factors.
2 Shading out the energy of insola-
tion by roofing or inert turbidity;
suppresses photosynthesis.
3 Introduction of substances toxic
to some fundamental part of the
food chain (such as copper sul-
phate) tends to temporarily inhibit
productivity.
SUMMARY
A A body of water such as a lake rep-
resents 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 mea-
sures to achieve a desired result.
REFERENCES
1 Chamberlin, Thomas C., and Salisburg,
Rollin P., Geology Vol. 1, "Geological
Processes and Their Results", pp i-xix,
and 1-654, Henry Holt and Company,
New York, 1904.
I 2-8
-------
OPTICS AND THE MICROSCOPE
I OPTICS
An understanding of elementary optics is
essential to the proper use of the microscope.
The microscopist will find that unusual pro-
blems in illumination and photomicrography
can be handled much more effectively once
the underlying ideas in physical optics are
understood.
A Reflection
A good place to begin is with reflection at
a surface or interface. Specular (or
regular) reflection results when a beam
of light leaves a surface at the same angle
at which it reached it. This type of
reflection occurs with highly polished
smooth surfaces. It is stated more pre-
cisely as Snell's Law, _i._e., the angle of
incidence, i, is equal to the angle of
reflection, r (Figure 1). Diffuse (or
scattered) reflection results when a beam
of light strikes a rough or irregular sur-
face and different portions of the incident
light are reflected from the surface at
different angles. The light reflected from
a piece of white paper or a ground glass is
an example of diffuse reflection.
Figure 1
SPECULAR REFLECTION - SNELL'S LAW
BI. MIC. 18.6.68
Strictly speaking, of course, all reflected
light, even diffuse, obeys Snell's Law.
Diffuse reflected light is made up of many
specularly reflected rays, each from a
a tiny element of surface, and appears
diffuse when the reflecting elements are
very numerous and very small. The terms
diffuse and specular, referring to reflection,
describe not so much a difference in the
nature of the reflection but rather a differ-
ence in the type of surface. A polished sur-
face gives specular reflection, a rough
surface gives diffuse reflection.
It is also important to note and remember
that specularly reflected light tends to be
strongly polarized in the plane of the reflect-
ing surface. This is due to the fact that
those rays whose vibration directions lie
closest to the plane of the reflection surface
are most strongly reflected. This effect is
strongest when the angle of incidence is
such that the tangent of the angle is equal
to the refractive index of the reflecting sur-
face. This particular angle of incidence is
called the Brewster angle.
B Image Formation on Reflection
Considering reflection by mirrors, we find
(Figure 2) that a plane mirror forms a
virtual image behind the mirror, reversed
right to left but of the same size as the
object. The word virtual means that the
image appears to be in a given plane but
that a ground glass screen or a photographic
film placed in that plane would show no
image. The converse of a virtual image is
a real image.
Spherical mirrors are either convex or con-
cave with the surface of the mirror repre-
senting a portion of the surface of a sphere.
The center of curvature is the center of the
sphere, part of whose surface forms the
mirror. The focus lies halfway between the
center of curvature and the mirror surface.
I 3-1
-------
Optics and the Microscope
Object
Virtual
Image
Mirror
Figure 2
IMAGE FORMATION BY PLANE MIRROR
Construction of an image by a concave
mirror follows from the two premises
given below (Figure 3)-
Figure 3
IMAGE FORMATION BY CONCAVE MIRROR
1 A ray of light parallel to the axis of
the mirror must pass through the
focus after reflection.
2 A ray of light which passes through the
center of curvature m-ist return along
the same path.
A corollary of the first premise is:
3 A ray of light which passes through the
focus is reflected parallel to the axis
of the mirror.
The image from an object can be located
using the familiar lens formula:
- + - 1
P q f
where p = distance from the object to
the mirror
q = distance from the image to
the mirror
f = focal length
C Spherical Aberration
No spherical surface can be perfect in its
image-forming ability. The most serious
of the imperfections, spherical aberration,
occurs in spherical mirrors of large
aperture (Figure 4). The rays of light
making up an image point from the outer
zone of a spherical mirror do not pass
through the same point as the more central
rays. This type of aberration is reduced by
blocking the outer zone rays from the image
area or by using aspheric surfaces.
Figure 4
SPHERICAL ABERRATION BY
SPHERICAL MIRROR
I 3-2
-------
Optic3 and the Microscope
D Refraction of Light
Turning now to lenses r.ithrr than nmtors
we find that the mo.sl impoi Unit i h.ir.n ler-
istic is refraction. Ki-Ti-.n linn ivli-is to
the change* ol direi lion .incl/oi veloi ily ol
light as it passes from one in diuin ID
another. The ratio of tin- \,ehn it\ in .in
(or more correi tlv in i\ v.u mini) lo tin-
velocity in the medium is i ailed the
refractive index. Som;' t.>pieal v.ilues ol
refractive index measureil with mono-
chromatic light (sodium D line) .ire listed
in Table 1.
Refraction causes an olijet t imnvrsed in
a medium of higher refractive index than
air to appear closer to the surfai e than it
actually is (Figure 5). This cffoc t may
into I'oi us and the new mu.rometer reading
is taken. Km.illy, the microscope is re-
rot used until the surface of the liquid appears
in sha i p foi us The micrometer reading
is Liken again and, with this information,
the refrac live index m?y be calculated from
the simplified equalion
. actual depth
index = - "- -
apparent depth
T.ihle I KKKRACT1VK INDICES OK COMMON
MATKRIA1.S MKASURKD WITH SODIUM LIGHT
Vac uum
Air
CO.,
Water
1.0000000
1 0002') 18
1. 0004498
1. VISO
Crown glass
Rock salt
Diamond
Lead sulfide
1.48 to
1.5443
2.417
3. 912
1. 61
Air
Actuol i
depth "
Apparent 1
depth \
\
Medium
image
i
.' Dhiarl
When the situation is reversed, and a ray
of light from a medium of high refractive
index passes through the interface of a
medium of lower index, the ray is refracted
until a critical angle is reached beyond which
all of the light is reflected from the interface
(Figure 6). This critical angle, C. has the
following relationship to the refractive indices
of the two media (
sin C
"9 L.
* , where
nl
-------
Optics and the Microscope
, E Dispersion
Dispersion is another important property
of transparent materials. This is the
variation of refractive index with color
(or wavelength) of light. When white light
passes through a glass prism, the light
rays are refracted by different amounts
and separated into the colors of the
spectrum. This spreading of light into
its component colors is due to dispersion
which, in turn, is due to the fact that the
refractive index of transparent substances,
liquids and solids, is lower for long wave-
lengths than for short wavelengths.
Because of dispersion, determination of
the refractive index of a substance re-
quires designation of the particular wave-
length used. Light from a sodium lamp
has a strong, closely spaced doublet with
an average wavelength of 5893A, called
the D line, which is commonly used as a
reference wavelength. Table 2 illustrates
the change of refractive index with wave-
length for a few common substances.
F Lenses
There arc two classes of lenses, con-
verging and diverging, called also convex
and concave, respectively. The focal
point of a converging lens is defined as
the point at which a bundle of light rays
parallel to the axis of the lens appears to
converge after passing through the lens.
The focal length of the lens is the distance
from the lens to the focal point (Figure 7).
Table 2. DISPERSION OF REFRACTIVE
INDICES OF SEVERAL COMMON MATERIALS
Refractive index
F line D line C line
blue (yellow) (red)
4861A 5893A 65631
Carbon disulfide
Crown glass
Flint glass
Water
1.
1.
1.
1.
6523
5240
6391
3372
1.6276
1.5172
1. 6270
1. 3330
1.
1.
1.
1.
6182
5145
6221
3312
The dispersion of a material can be defined
quantitatively as:
n (yellow) - 1
v = dispersion
n (blue) - n (red)
n (593mji) - 1
n (486mn) -
where n is the refractive index of the
material at the particular wavelength
noted in the parentheses.
Figure 7
CONVERGENCE OF LIGHT AT FOCAL POINT
G Image Formation by Refraction
Image formation by lenses (Figure 8)
follows rules analogous to those already
given above for mirrors
1 Light traveling parallel to the axis of
the lens will be refracted so as to pass
through the focus of the lens.
2 Light traveling through the geometrical
center of the lens will be unrefracted.
The position of the image can be determined
by remembering that a light ray passing
through the focus, F, will be parallel to
the axis of the lens on the opposite side of
the lens and that a ray passing through the
geometrical center of the lens will be
unrefracted.
I 3-4
-------
Optics and the Microscope
Figure 8
IMAGE FORMATION BY A CONVEX LENS
The magnification, M, of an image of an
object produced by a lens is given by the
relationship-
image size _ image distance _ q
object size object distance p
where q = distance from image to lens
and p = distance from object to lens.
H Aberrations of Lenses
Lenses have aberrations of several types
which, unless corrected, cause loss of
detail in the image. Spherical aberration
appears in lenses with spherical surfaces.
Reduction of spherical aberration can be
accomplished by diaphragming the outer
zones of the lens or by designing special
aspherical surfaces in the lens system.
Chromatic aberration is a phenomenon
caused by the variation of refractive index
with wavelength (dispersion). Thus a lens
receiving white light from an object will
form a violet image closer to the lens and
a red one farther away. Achromatic
lenses are employed to minimize this
effect. The lenses are combinations of
two or more lens elements made up of
materials having different dispersive
powers. The use of monochromatic light
is another obvious way of eliminating
chromatic aberration.
Astigmatism is a third aberration of
spherical lens systems It occurs when
object points arc- not located on the optical
axis of the lens and results in the formation
of an mclistiml image. The simplest
remedy for astigmatism is to place the
ohjei I (lose to the axif, of the lens system.
Interfere e Phenomena
Intcrfcrem c and diffraction are two phe-
nomena whieh arc due to the wave character-
istics of light. The superposition of two
light rays arriving simultaneously at a given
point will give rise to interference effects,
whereby the intensity at that point will vary
from dark to bright depending on the phase
different cs between the two light rays.
The first requirement for interference is
that the light must come from a single
source. The light may be split into any
number of paths but must originate from
the same point (or coherent source). Two
light waves from a coherent source arriv-
ing at a point in phase agreement will
reinforce each other (Figure 9a). Two
light waves from a coherent source arriv-
ing at a point in opposite phase will cancel
each other (Figure 9b).
Figure 9a. Two light rays, 1 and 2, of
the same frequency but dif-
ferent amplitudes, are in phase
in the upper diagram. In the
lower diagram, rays 1 and 2
interfere constructively to give
a single wave of the same fre-
quency and with an amplitude
equal to the summation of the
two former waves.
I 3-5
-------
jtics and the Microscope
otb
Figure 9b. Rays 1 and 2 are now 180°
out of phase and interfere
destructively. The resultant,
in the bottom diagram, is of
the same frequency but is of
reduced amplitude (a is
negative and is subtracted
from b).
The reflection of a monochromatic light
beam by a thin film results in two beams,
one reflected from the top surface and one
from the bottom surface. The distance
traveled by the latter beam in excess of
the first.is twipe the thickness of the film
and its equivalent air path is:
2 nt
where n is the refractive index and
t is the thickness of the film.
The second beam, however, upon reflection
at the bottom surface, undergoes a half
wavelength shift and now the total retard-
ation of the second beam with respect to
the first is given as:
retardation = 2 nt +
where \ is the wavelength of the light
beam.
When retardation is exactly an odd number
of half wavelengths, destructive interfer-
ence takes place resulting in darkness.
When it is zero or an even number of half
wavelengths, constructive interference
results in brightness (Figure 10).
I 3-6
Figure 10
INTERFERENCE IN A THIN FILM
A simple interferometer can be made by
partially silvering a microscope slide and
cover slip. A preparation between the two
partially silvered surfaces will show inter-
ference fringes when viewed with mono-
chromatic light, either transmitted or by
vertical illuminator. The fringes will be
close together with a wedge-shaped prep-
aration and will reflect refractive index
differences due to temperature variations,
concentration differences, different solid
phases, etc. The method has been used to
measure quantitatively the concentration of
solute around a growing crystal'^(Figure 11).
,50% Mirror/
N
Cover slip- -
Specimen
x-100%
/ Mirror
Figure 11
MICROSCOPICAL METHOD OF VIEWING
INTERFERENCE IMAGES
a Examination is by transmitted light.
Light ray undergoes multiple
reflections and produces dark and
light fringes in the field.. A speci-
men introduces a phase shift and
changes the fringe pattern.
b Illumination is from the top. The
principle is the same but fringes
show greater contrast.
-------
Optics and the Microscope
Each dark band represents an equivalent
air thickness of an odd number of half
wavelengths. Conversely, each bright
band is the result of an even number of
half wavelengths.
With interference illumination, the effect
of a transparent object of different re-
fractive index than the medium in the
microscope field is:
1 a change of light intensity of the object
if the background is uniformly illumi-
nated (parallel cover slip), or
2 a shift of the interference bands within
the object if the background consists
of bands (tilted cover slip).
The relationship of refractive indices of
the surrounding medium and the object is
as follows:
ns= nm(l
d =
2.44 fX
where f is the focal length of the lens,
X the wavelength, and D the diameter
of the lens.
It is seen that in order to maintain a
small diffraction disc at a given wave-
length, the diameter of the lens should
be as large as possible with respect to
the focal length. It should be noted,
also, that a shorter wavelength produces
a smaller disc.
If two pin points of light are to be distin-
guished in an image, their diffraction discs
must not overlap more than one half their
diameters. The ability to distinguish such
image points is called resolving power and
is expressed as one half of the preceding
expression
1.22 f X
resolving power = .
where ns
X
t
J Diffraction
refractive index of the
specimen
refractive index of the
surrounding medium
phase shift of the two
beams, degrees
wavelength of the light
thickness of the specimen.
In geometrical optics, it is assumed that
light travels in straight lines. This is not
always true. We note that a beam passing
through a slit toward a screen creates a
bright band wider than the slit with alter-
nate bright and dark bands appearing on
either side of the central bright band,
decreasing in intensity as a function of
the distance from the center. Diffraction
describes this phenomenon and, as one of
its practical consequences, limits the
lens in its ability to reproduce an image.
For example, the image of a pin point of
light produced by a lens is not a pin point
but is revealed to be a somewhat larger
patch of light surrounded by dark and
bright rings. The diameter, d, of this
diffraction disc (to the first dark ring)
is given as:
II THE COMPOUND MICROSCOPE
The compound microscope is an extension in
principle of the simple magnifying glass,
hence it is essential to understand fully the
properties of this simple lens system.
A Image Formation by the Simple Magnifier
The apparent size of an object is determined
by the angle that is formed at the eye by the
extreme rays of the object. By bringing the
object closer to the eye, that angle (called
the visual angle) is increased. This also
increases the apparent size. However a
limit of accommodation of the eye is reached,
at which distance the eye can no longer focus.
This limiting distance is about 10 inches or 25
centimeters. It is at this distance that the
magnification of an object observed by the
unaided eye is said to be unity. The eye can,
of course, be focused at shorter distances but
not usually in a relaxed condition.
A positive, or converging, lens can be used
to permit placing an object closer than 10
inches to the eye (Figure 12). By this means
the visual angle of the object is increased
(as is its apparent size) while the image of
I 3-7
-------
Optics and the Microscope
Eye
I mog»
Mognifitr
Figure 12
VIRTUAL IMAGE FORMATION BY
CONVEX LENS
the object appears to be 10 inches from
the eye, where it is best accommodated.
B Magnification by a Single Lens System
The magnification, M, of a simple magni-
fying glass is given by:
M= f +1
where f = focal length of the lens in
centimeters.
Theoretically the magnification can be
increased with shorter focal length lenses.
However such lenses require placing the.
eye very close to the lens surface and
have much image distortion and other
optical aberrations. The practical limit
for a simple magnifying glass is about
2 OX.
In order to go to magnifications higher
than 20X, the compound microscope is
required. Two lens systems are used
to form an enlarged image of an object
(Figure 13). This is accomplished in
two steps, the first by a lens called the
objective and the second by a lens known
as the eyepiece (or ocular).
C The Objective
The objective is the lens (or lens system)
closest to the object. Its function is to
reproduce an enlarged image of the object
in the body tube of the microscope.
Objectives are available in various focal
Eyepiece
Objective
Victual
Figure 13
IMAGE FORMATION IN
COMPOUND MICROSCOPE
lengths to give different magnifications
(Table 3). The magnification is calculated
from the focal length by dividing the latter
into the tube length, usually 160 mm.
The numerical aperture (N. A.) is a measure
of the ability of an objective to resolve detail.
This is more fully discussed in the next
section. The working distance is in the free
space between the objective and the cover
slip and varies slightly for objectives of the
same focal length depending upon the degree
of correction and the manufacturer.
There are three basic classifications of
objectives: achromats, fluorites and
apochromats, listed in the order of their
complexity. The achromats are good for
routine work while the fluorites and apo-
chromats offer additional optical corrections
to compensate for spherical, chromatic and
other aberrations.
I 3-8
-------
Table 3. NOMINAL CHARACTERISTICS OK USUAL
Nominal
focal length
mm
56
32
16
8
4
4
1.8
Nominal
magnif.
2.5X
5
10
20
43
45
90
N.A.
0.08
0. 10
0.25
0.50
0. 66
0.85
1.30
Working
distance
mm
40
25
7
1. 3
0. 7
0.5
0.2
Depth
focus
V-
50
16
8
2
1
1
0.4
Diam. of
field
mm.
8.5
5
2
1
0.5
0.4
0.2
MICROSCOPE OBJECTIVES
Resolving
power, white
light. K
4.4
3.9
1.4
0.7
0.4
0.35
0.21
Maximum
useful
magnif.
SOX
90X
250X
500X
660X
850X
1250X
Eyepiece
for max.
useful magnif.
30X
20X
25X
25X
15X
20X
12X
Another system of objectives employs
reflecting surfaces in the shape of concave
and convex mirrors. Reflection optics,
because they have no refracting elements,
do not suffer from chromatic aberrations
as ordinary refraction objectives do. Based
entirely on reflection, reflecting objectives
are extremely useful in the infrared and
ultraviolet regions of the spectrum. They
also have a much longer working distance
than the refracting objectives.
The body tube of the microscope supports
the objective at the bottom (over the object)
and the eyepiece at the top. The tube
length is maintained at 160 mm except for
Leitz instruments, which have a 170-mm
tube length.
The objective support may be of two kinds,
an objective clutch changer or a rotating
nosepiece
1 The objective clutch changer ("quick-
change" holder) permits the mounting ,
of only one objective at a time on the
microscope. It has a centering arrange-
ment, so that each objective need be
centered only once with respect to the
stage rotation. The changing of objec-
tives with this system is somewhat
awkward compared with the rotating
nosepiece. -
2 The revolving nosepiece allows mounting
three or four objectives on the microscope
at one time (there are some nosepieces
that accept five and even six objectives).
In this system, the objectives are
usually noncenterable and the stage is
centerable. Several manufacturers pro-
vide centerable objective mounts so that
each objective on the nosepiece need be
centered only once to the fixed rotating
stage. The ins ides of objectives are
better protected from dust by the rotating
nosepiece. This, as well as the incon-
venience of the so-called "quick-change"
objective holder, makes it worthwhile
to have one's microscope fitted with
rotating nosepiece.
D The Ocular
The eyepiece, or ocular, is necessary in
the second step of the magnification process.
The eyepiece functions as a simple magni-
fier viewing the image formed by the
objective.
There are three classes of eyepieces in
common use- huyghenian, compensating
and flat-field. The huyghenian (or huyghens)
eyepiece is designed to be used with
achromats while the compensating type is
used with fluorite and apochromatic
objectives. Flat-field eyepieces, as the
name implies, are employed in photo-
micrography or projpction and can be used
with most objectives. It is best to follow
the recommendations of the manufacturer
as to the proper combination of objective
and eyepiece.
I 3-9
-------
Optics and the Microscope
The usual magnifications available in
oculars run from about 6X up to 25 or
SOX. The 6X is generally too low to be of
any real value while the 25 and 30X oculars
have slightly poorer imagery than medium
powers and have a very low eyepomt. The
most useful eyepieces lie in the 10 to 20X
magnification range.
E Magnification of the Microscope
The total magnification of the objective-
eyepiece combination is simply the product
of the two individual magnifications. A
convenient working rule to assist in the
proper choice of eyepieces states that the
maximum useful magnification (MUM) for
the microscope is 1, 000 times the numeri-
cal aperture (N.A.) of the objective.
The MUM is related to resolving power
in that magnification in excess of MUM
gives little or no additional resolving
power and results in what is termed empty
magnification. Table 4 shows the results
of such combinations and a comparison
with the 1000XN. A. rule. The under-
lined figure shows the magnification near-
est to the MUM and the eyepiece required
with each objective to achieve the MUM.
From this table it is apparent that only
higher power eyepieces can give full use
of the resolving power of the objectives.
It is obvious that a 10X. or even a 15X,
eyepiece gives insufficient magnification
for the eye to see detail actually resolved
by the objective.
F Focusing the Microscope
The coarse adjustment is used to roughly
position the body tube (in some newer
microscopes, the stage) to bring the image
into focus. The fine adjustment is used
after the coarse adjustment to bring the
image into perfect focus and to maintain
the focus as the slide is moved across the
stage. Most microscope objectives are
parfocal so that once they are focused any
other objective can be swung into position
without the necessity of refocusing except
with the fine adjustment.
The student of the microscope should first
learn to focus in the following fashion, to
prevent damage to a specimen or objective:
1 Raise the body tube and place the speci-
men on the stage.
2 Never focus the body tube down (or the
stage up) while observing the field
through the eyepiece.
3 Lower the body tube (or raise the stage)
with the coarse adjustment while care-
fully observing the space between the
Table 4. MICROSCOPE MAGNIFICATION CALCULATED
FOR VARIOUS OBJECTIVE-EYEPIECE COMBINATIONS
Objective
Focal Magni-
length
56mm
32
16
8
4
1.8
fication
3X
5
10
20
40
90
5X
15X
25X
SOX
100X
200X
450X
10X
30X
50X
100X
200X
400X
900X
Eyepiece
15X
45X
75X
150X
300X
600X
1350X
20X
60X
100X
200X
40 OX
800X
1800X
25X
75X
125X
250X
500X
1000X
2250X
MUMa
(1000 NA)
SOX
100X
250X
50 OX
660X
1250X
aMUM = maximum useful magnification
I 3-10
-------
Optic3 and the Microscope
objertivr and shclo and permitting the
two to come close together without
touching.
4 Looking through the microscope and
turning the fine- adjustment in such a
way as to move the objective away from
the specimen, bring the image into
sharp focus.
The fine adjustment is usually calibrated
in one- or two-micron steps to indicate
the vertical movement of the body tube.
This feature is useful in making depth
measurements but should not be relied
upon for accuracy.
G The Substage Condenser
The substage holds the condenser and
polarizer. It can usually be focused in a
vertical direction so that the condenser can
be brought into the correct position with
respect to the specimen for proper
illumination. In some models, the conden-
ser is centerable so that it may be set
exactly in the axis of rotation of the stage,
otherwise it will have been precentered at
the factory and should be permanent.
H The Microscope Stage
The stage of the microscope supports the
specimen between the condenser and
objective, and may offer a mechanical stage
as an attachment to provide a means of
moving the slide methodically during obser-
vation. The polarizing microscope is
fitted with a circular rotating stage to
which a mechanical stage may be added.
The rotating stage, which is used for object
orientation to observe optical effects, will
have centering screws if the objectives are
not centerable, or vice versa. It is un-
desirable to have both objectives and stage
centerable as this does not provide a fixed
reference axis.
I The Polarizing Elements
A polarizer is fitted to the condenser of all
polarizing microscopes. In routine instru-
ments, the polarizer is fixed with its
vibration direction oriented north-south
(east-west for most European instruments)
while in research microscopes, the
polarizer can be rotated. Modern instru-
ments have polarizing filters (such as
Polaroid) replacing the older calcite
prisms. Polarizing filters are preferred
because they
1 are low-cost,
2 require no maintenance,
3 permit use of the full condenser
aperture
An analyzer, of the same construction as
the polarizer, is fitted in the body tube of
the microscope on a slider so that it may
be easily removed from the optical path.
It is oriented with its plane of vibration
perpendicular to the corresponding direction
of the polarizer.
J The Bertrand Len»
The Bertrand lens is usually found only on
the polarizing microscope although some
manufacturers are beginning to include it
on phase microscopes. It is located in the
body tube above the analyzer on a slider
(or pivot) to permit quick removal from
the optical path. The Bertrand lens is used
to observe the back focal plane of the objective.
It is convenient for checking quickly the type
and quality of illumination, for observing
interference figures of crystals, for adjust-
ing the phase annuli in phase microscopy
and for adjusting the annular and central
stops in dispersion staining.
K The Compensator Slot
The compensator slot receives compensators
(quarter-wave, first-order red and quartz-
wedge) for observation of the optical prop-
erties of crystalline materials. It is usually
placed at the lower end of the body tube just ,
above the objective mount, and is oriented
45° from the vibration directions of the
polarizer and analyzer.
L The Stereoscopic Microscope
The stereoscopic microscope, also called
the binocular, wide-fie Id, dissecting or
I 3-11
-------
Optics and the Microscope
Greenough binocular microscope, is in
reality a combination of two separate
compound microscopes. The two micro-
scopes, usually mounted in one body, have
their optical axes inclined from the vertical
by about 7 and from each other by twice
this angle. When an object is placed on the
stage of a stereoscopic microscope, the
optical systems view it from slightly
different angles, presenting a stereoscopic
pair of images to the eyes, which fuse the
two into a single three-dimensional image.
The objectives are supplied in pairs, either
as separate units to be mounted on the
microscope or, as in the new instruments,
built into a rotating drum. Bausch and
Lomb was the first manufacturer to have a
zoom lens system which gives a continuous
change in magnification over the full range.
Objectives for the stereomicroscope run
from about 0. 4X to 12X, well below the
magnification range of objectives available
for single-objective microscopes.
The eyepieces supplied with stereoscopic
microscopes run from 10 to 25X and have
wider fields than their counterparts in the
single-objective microscopes.
Because of mechanical limitations, the
stereomicroscope is limited to about 200X
magnification and usually does not permit
more than about 120X. It is most useful
at relatively low powers in observing
shape and surface texture, relegating the
study of greater detail to the monocular
microscope. The stereomicroscope is
also helpful in manipulating small samples,
separating ingredients of mixtures, pre-
paring specimens for detailed study at
higher magnifications and performing
various mechanical operations under micro-
scopical observation, e. g. micromampulation.
Ill ILLUMINATION AND RESOLVING POWER
Good resolving power and optimum specimen
contrast are prerequisites for good microscopy.
Assuming the availability of suitable optics
(ocular, objectives and substage condenser)
it is still of paramount importance to use
proper illumination. The requirement for a
good illumination system for the microscope
is to have uniform intensity of illumination
over the entire field of view with independent
control of intensity and of the angular aperture
of the illuminating cone.
A Basic Types of Illumination
There are three types of illumination
(Table 5) used generally.
1 Critical. This is used when high levels
of illumination intensity are necessary
for oil immersion, darkfield, fluores-
cence, low birefringence or photo-
micrographic studies. Since the lamp
filament is imaged in the plane of the
specimen, a ribbon filament or arc
lamp is required. The lamp must be
focusable and have an iris diaphragm,
the position of the filament must also
be adjustable in all directions.
2 Kohler. Also useful for intense illumi-
nation, Kohler illumination may be
obtained with any lamp not fitted with a
ground glass. The illuminator must,
however, be focusable, it must have an
adjustable field diaphragm (iris) and the
lamp filament position must be adjust-
able in all directions.
3 "Poor man's". So-called because a low-
priced illuminator may be used, this
method gives illumination of high quality
although of lower intensity because of the
presence of a ground glass in the system.
No adjustments are necessary on the
illuminator or lamp filament although
an adjustable diaphragm on the illuminator
is helpful.
All three types of illumination require that
the microscope substage condenser focus
the image of the illuminator aperture in the
plane of the specimen. In each case, then,
the lamp iris acts as a field diaphragm and
should be closed to just illuminate the field
of view. The differences in these three
types of illumination lie in the adjustment
of the lamp condensing lens. With poor
man's illumination there is no lamp conden-
ser, hence no adjustment. The lamp should
be placed close to the microscope so that
I 3-12
-------
Optics and the Microscope
Table 5. COMPARISON OF CRITICAL.
KOHLER AND JfCOR MAN'S ILLUMINATION
Lamp filament
Lamp condensing lens
Lamp iris
Ground glass at lamp
Image of light source
Image of field iris
Image of substage iris
Critical
ribbon filament
required
required
none
m object plane
near object
plane
back focal plane
of objective
Kohler
any type
required
required
none
at substage
iris
in object
plane
back focal plane
of objective
Poor man's
any type
none
useful
present
none
near object
plane
back focal plane
of objective
the entire field of view is always
illuminated. If the surface structure of the
ground glass becomes apparent in the field
of view the subs:age condenser is very
slightly defocused.
Critical Illumination
With critical illumination the lamp conden-
ser is focused to give parallel rays; focus-
ing the lamp filament on a far wall is
sufficient. Aimed, then, at the substage
mirror, the substage condenser will focus
the lamj, filament in the object plane. The
substage condenser iris will now be found
imaged in the back focal plane of the ob-
jective; it serves as a control over con-
vergence of the illumination. Although the
substage iris also affects the light intensity
over the field of view it should most decid-
edly not be used for this purpose. The
intensity of illumination may be varied by
the use of neutral density filters and. unless
color photomicrography is anticipated, by
the use of variable voltage on the lamp
filament.
Kohler illumination (Figure 14) differs
from critical illumination in the use of the
lamp condenser. With critical illumination
the lamp condenser focuses, the lamp
filament at infinity, with Kohler illumination
the lamp filament is focused in the plane of
the substage condenser iris (also coincident
with the anterior focal plane of the substage
condenser). The functions of the lamp
condenser iris and the substage condenser
iris in controlling, respectively, the area
of the illuminated field of view and the
angular aperture of the illuminating cone
are precisely alike for all three types of
illumination.
Critical illumination is seldom used because
it requires a special lamp filament and be-
cause, when used, it shows no advantage
over well-adjusted Kohler illumination.
n
Kohler Illumination
To arrange the microscope and illuminator
for Kohler illumination it is well to proceed
through the following steps:
a Remove the diffusers and filters
from the lamp.
b Turn the lamp on and aim at a con-
venient wall or vertical screen about
19 inches away. Open the lamp
diaphragm.
c By moving the lamp condenser, focus
a sharp image of the filament. It
should be of such a size as to fill,
not necessarily evenly, the microscope
I 3-13
-------
Optics and the Microscope
Critical
Kohler
Focal .plane
Poor man's
Objective
Preparation
Substaqe
condenser
Substage j
iris
Lamp iris
Lamp
condenser
Light source
I 3-14
-------
Opties and the Microscope
substagc condenser opening. II il
docs not, move the lamp aw.ix I nun
the wall to enlarge the I'll.inicnl inuj;r,
refocus.
d Turn the lamp JIN! .inn il .it Ihe mu 111-
scope mirroi t»o a* to m.iml.i in the
same 18 inches (01 atljusled l.imp
distance).
e Place a specimen on the mil i cisi ope
stage and foeiib .sluirplx. with a Id-mm
(10X) objective. Open fully the
aperture diaphragm in the substage
condenser. If the light is too bright,
temporarily place a neutral density
filter or a diffuser in the lamp.
f Close the lamp diaphragm, or field
diaphragm, to about a 1-cm opening.
Rack the microscope substage con-
denser up and down to focus the
field diaphragm sharply in the same
plane as the specimen.
g Adjust the mirror to center the field
diaphragm in the field of view.
h Remove the 16-mm objective and
replace with a 4-mm objective. Move
the specimen so that a clear area is
under observation. Place the
Bertrand lens in the optical path, or
remove the eyepiece and insert an
auxiliary telescope (sold with phase
contrast accessories) in its place.
or remove the eyepiece and observe
the back aperture of the objective
directly. Remove any ground glass
diffusers from the lamp. Now
observe the lamp filament through
the microscope.
i If the filament does not appear to be
centered, swing the lamp housing in
a horizontal arc whose center is at
the field diaphragm. The purpose
is to maintain the field diaphragm on
the lamp in its centered position. If
a vertical movement of the filament
is required, loosen the bulb base and
slide it up or down. If the base is
fixed, tilt the lamp housing in a
vertical arc with the field diaphragm
.is tin- i enter of movement (again
enile.ivormg to Keep the lamp dia-
plitM^ni in tht t entered position).
II you have- mistered this step, you
ll.ivr ac (umplished the most d iff H Ult
|)()ilioii (Heller mil lost Ope lamps
h.ivc .id iiislmcnls lo move the bulb
independently ul the la mo housing to
simplify this step.)
I ljul the specimen in plate, replace
the eyepiei e and the desired ob|ec-
tjvc ,md relot us.
k Open or i lose the field diaphragm
until it just disappears from the field.
1 Observe the back aperture of the
objective, preferably with the Bertrand
lens or the.auxiliary telescope, and
close the aperture diaphragm on the
substage condenser until it is about
four-fifths the diameter of the back
aperture. This is the best position
for the aperture diaphragm, a posi-
tion which minimizes glare and maxi-
mizes the resolving power. It is
instructive to vary the aperture dia-
phragm and observe the image criti-
cally during the manipulation.
m If the illumination is too great,
insert an appropriate neutral density
filter between the illuminator and
the condenser. Do not use the con-
denser aperture diaphragm or the
lamp field diaphragm to control the
intensity of illumination.
Poor Man's Illumination
Both critical and Kohler illumination re-
quire expensive illuminators with adjust-
able focus, lamp iris and adjustable lamp
mounts. Poor man's illumination requires
a cheap illuminator although an expensive
illuminator may be used if its expensive
features are negated by inserting a ground
glass diffuser or by using a frosted bulb.
Admittedly an iris diaphragm on the lamp
would be a help though it is not necessary.
a The illuminator must have a frosted
bulb or a ground glass diffuser.
I 3-15
-------
Optics and the Microscope
It should be possible to direi t it in
the general direction of the substage
mirror, very close thereto 01 in
place thereof.
b Focus on any preparation after
tilting the mirror to illuminate the
field.
c Remove the top lens ol the londenser
and, by racking the i ondenser up or,
more often, down, bring into foe us
(in the same plane as the spei iirn>n)
a finger, pencil or other objei t plat ed
in the same general region at> the
ground glass diffuser on the- lamp.
The glass surface itself ian then be
focused in the plane of the spei invn.
d Ideally the ground glass surface will
just fill the field of view when centered
by the substage mirror, adjustment
may be made by moving the lamp
closer to or farther from the micro-
scope (the position might be marked
for each objective used) or by cutting
paper diaphragms of fixed aperture
(one for each objective used). In this
instance a lamp iris would be useful.
e Lower the condenser just sufficiently
to defocus the ground glass surface
and render the field of illumination
even.
f Observe the back aperture of the
objective and open the substage con-
denser iris about 75 percent of the
way. The final adjustment of the
substage iris is made while observing
the preparation, the iris should be
open as far as possible, still giving
good contrast.
g The intensity of illumination should
be adjusted only with neutral density
filters or by changing the lamp voltage.
Proper illumination is one of the most im-
portant operations in microscopy. It is
easy to judge a microscopist's ability by
a glance at his field of view and the objec-
tive back lens.
I 3-16
H Kesolvmg Power
The resolving power of the microscope is
its .iluhly U> distinguish separate details
of i losely spai ed microscopic structures.
The Iheoreln al limit of resolving two
(list rele points, a distant e X apart, is
x TS.T.
where \ - wavelength of light used to
illuminate the specimen
N.A. = numerical aperture of the
objective
Substituting a wavelength of 4, 500
Angstroms and a numerical aperture of
1. i, about the best that can be done with
visible light, we find that two points about
2, OOOA (or 0. 2 micron) apart can be seen
as two separate points. Further increase
in resolving power can be achieved for the
light mic roscope by using light or shorter
wavelength. Ultraviolet light near 2, 000
Angstroms lowers the limit to about 0. 1
micron, the lower limit for the light
microscope.
The numerical aperture of an objective is
usually engraved on the objective and is
related to the angular aperture, AA
(Figure 15), by the formula:
N.A. = n sin
AA
2
where n = the lowest index in the space
between the object and the
objective.
Angular aperture
Object
Figure 15
ANGULAR APERTURE OF
MICROSCOPE OBJECTIVE
-------
Optics and the Microscope
1 Maximum useful magnification
A helpful rule of thumb is that the use-
ful magnification will not exceed 1, 000
times the numerical aperture of the
objective (see Tables 3 and 4). Although
somewhat higher magnification may be
used in specific cases, no additional
detail will be resolved.
It is curious, considering the figures
in the table, that most, if not all, manu-
facturers of microscopes furnish a 10X
eyepiece as the highest power. A 10X
eyepiece is useful but anyone interested
in critical work should use a 15-25X eye-
piece; the 5-10X eyepieces are best for
scanning purposes.
2 Abbe's theory of resolution
One of the most cogent theories of
resolution is due to Ernst Abbe, who
suggested that microscopic objects act
like diffraction gratings (Figure 16) and
that the angle of diffraction, therefore,
increases with the fineness of the detail.
He proposed that a given microscope
objective would resolve a particular
detail if at least two or three transmitted
rays (one direct and two diffracted rays)
entered the objective. In Figure 16 the
detail shown would be resolved in A and
C but not in B. This theory, which can
be borne out by simple experiment, is
useful in showing how to improve resolu-
tion. Since shorter wavelengths will
give a smaller diffraction angle, there
is more chance of resolving fine detail
with short wavelengths. Also, since
only two of the transmitted rays are
needed, oblique light and a high N. A.
condenser will aid in resolving fine detail.
3 Improving resolving power
The following list summarizes the
practical approaches to higher resolu-
tion with the light microscope:
a The specimen should be illuminated
by either critical or Kohler
illumination.
/
Figure 16
ABBE THEORY OF RESOLUTION
b The condenser should be well-
corrected and have a numerical
aperture as high as the objective to
be used.
c An apochromatic oil-immersion
objective should be used with a com-
pensating eyepiece of at least 15X
magnification. The immersion oil
should have an index close to 1. 515
and have proper dispersion for the
objective being used.
d Immersion oil should be placed
between the condenser and slide and
between cover slip and objective.
The preparation itself should be
surrounded by a liquid having a
refractive index of 1.515 or more.
e The illumination should be reasonably
monochromatic and as short in wave-
length as possible. An interference
filter transmitting a wavelength of
about 480-500 millimicrons is a
suitable answer, to this problem.
Ideally, of course, ultraviolet light
should be used to decrease the wave-
length still further.
The practical effect of many of these
factors is critically discussed by
Loveland(2) in a paper on the optics of
object space.
I 3-17
-------
Optics and the Microscope
IV PHOTOMICROGRAPHY
A Introduction
Photomicrography, as distinct from micro-
photography, is the art of taking pictures
through the microscope. A microphoto-
graph is a small photograph, a photomicro-
graph is a photograph of a small object.
Photomicrography is a valuable tool in
recording the results of microscopical
study. It enables the microscopist to
1 describe a microscopic field objectively
without resorting to written descriptions,
2 record a particular field for future
reference,
3 make particle size counts and counting
analyses easily and without tying up a
microscope,
4 enhance or exaggerate the visual micro-
scopic field to bring out or emphasize
certain details not readily apparent
visually,
5 record images in ultraviolet and infra-
red microscopy which are otherwise
invisible to the unaided eye.
There are two general approaches to photo-
micrography, one requires only a plate or
film holder supported above the eyepiece
of the microscope with a light-tight bellows,
the other utilizes any ordinary camera with
its own lens system, supported with a light-
tight adaptor above the eyepiece. It is
best, in the latter case, to use a reflex
camera so that the image can be carefully
focused on the ground glass. Photomi-
crography of this type can be regarded
simply as replacing the eye with the camera
lens system. The camera should be focused
at infinity, just as the eye is for visual
observation, and it should be positioned
close to and over the eyepiece.
The requirements for photomicrography,
however, are more rigorous than those
for visual work. The eye can normally
compensate for varying light intensities,
curvature of field and depth of field. The
photographic plate, however, lies in one
plane, hence the greatest care must be
used to focus sharply on the subject plane
of interest and to select optics to give
minimum amounts of field curvature and
chromatic aberrations.
With black and white film, color filters
may be used to enhance the contrast of
some portions of the specimen while mini-
mizing chromatic aberrations of the lenses.
In color work, however, filters cannot
usually be used for this purpose and better
optics may be required.
Photomic-rographic cameras which fit
directly onto the microscope are available
in 35-mm or up to 3-1/4 X 4-1/4 inch sizes.
Others are made which accommodate larger
film sizes and which have their own support
independent of the microscope. The former,
however, are preferred for ease of handling
and lower cost. The latter system is pre-
ferred for greater flexibility and versatility
and lack of vibration. The Polaroid camera
has many applications in microscopy and
can be used on the microscope directly but,
because of its weight, only when the micro-
scope has a vertically moving stage for
focusing rather than a focusing body tube.
B Determination of Correct Exposure
Correct exposure determination can be
accomplished by trial and error, by relating
new conditions to previously used successful
conditions and by photometry.
With the trial and error method a series of
trial exposures is made, noting the type of
subject, illumination, filters, objective,
eyepiece, magnification, film and shutter
speed. The best exposure is selected. The
following parameters can be changed and
the exposure time adjusted accordingly:
1 Magnification. Exposure time varies
as the square of the magnification.
Example Good exposure was obtained
with a 1/10-second exposure
and a magnification of 100X.
If the magnification is now
I 3-18
-------
Optics and the Microscope
200X, the correct exposure
is calculated as follows-
new exposure time = old exposure time
.new magnification.2 . ,200.2
lold magnification ' ' M00;
4/10 or, say, 1/2 second.
Kodachrome II Type A
Professional is 40.
new exposure time = old exposure time
x A. S. A. of old film = 1/100(400/40) ,
A. S. A. of new film
10/100 or 1/10 second.
It should be noted, however, that the
above calculation can be made only when
there has been no change in the illumi-
nation system including the condenser
or the objective. Only changes in magni-
fication due to changing eyepieces or
bellows extension distance can be hand-
led in the above manner.
2 Numerical aperture. Exposure time
varies inversely as the square of the
smallest working numerical aperture
of the condenser and objective.
Example: Good exposure was obtained
at 1/10 second with the 10X
objective, N.A. 0.25, at
full aperture. With a 20X
objective, N.A. 0.25, at
full aperture and the same
final magnification, what is
the correct exposure time?
new exposure time = old exposure time
/\ i^"^^^»^^^r"~ *
new N.A.
say, 1/50 second.
= 1/40 or,
It is seen that more light reaches the
photographic film with higher numeri-
cal apertures at the same magnification.
3 Film. Exposure time varies inversely
with the American Standards Association
speed index of the film.
Example: A good picture was obtained
with Eastman Tn-X film at
I/100 second. What is the
correct exposure for
Eastman Kodachrome II
Type A. The A. S. A. speed
for Tri-X is 400 and for
4 Other parameters may be varied but the
prediction of exposure time cannot be
made readily. Experience and photo-
electric devices are the best guides to
the proper exposure.
Photoelectric devices are excellent for
determining correct exposure. Since
ordinary photographic exposure meters
are not sensitive enough for photomi-
crography, more sensitive instruments,
having a galvanometer or electronic
amplifying circuit, are required. Some
photosensitive cells are inserted in the
body tube in place of the eyepiece for
light intensity readings. This has the
advantage of detecting the light level at a
point of high intensity but does not take
into account the eyepiece, the distance to
the film or the film speed.
The cell may be placed just above the eye-
piece so that it registers the total amount
of light leaving the eyepiece. Again, the
effects of film speed and the projection
distance are not accounted for. The prin-
cipal drawback with the total light
measuring photometer is the difficulty of
taking into account the area of field covered.
Take, for example, a bright field in which
only a few crystals appear, perhaps 1 per-
cent of the light entering the field of view is
scattered by the crystals and the photometer
shows close to a maximum reading. Now
assume that everything remains constant
except the number of crystals and. conse-
quently, the amount of light scattered.
The photometer reading could easily drop
by 50 percent, yet the proper exposure is
unchanged. The situation is similar for
photomicrography with crossed polars since
the photometer reading depends on the
intensity of illumination, on the bire-
fringence and thickness of the crystals and
I 3-19
-------
Optics and the Microscope
on the number and size of the crystals in
the field or, alternatively, on the area of
the field covered by birefringcnt crystals.
One of the best solutions to this problem
is to measure the photometer reading with
no preparation on the stage. A first-order
red compensator or a quartz wedge is in-
serted when crossed polars are being used
to illuminate the entire field.
An alternative is to place the cell on the
ground glass where the film will be
located. However, although all variables
except film speed are now taken into
account, measurements in the image plane
have the disadvantage of requiring a more
sensitive electronic photoelectric apparatus.
No matter what method is used for placing
the photocell, the exposure time can be
determined by the general formula
exposure time =
meter reading
The constant k will depend on the physical
arrangement and film used. To determine
k for any particular system, first set up
the microscope to take a picture. Record
the meter reading and take a series of
trial exposures. Pick out the best exposure
and calculate k. Then the k which was
determined holds as long as no change is
made in the light path beyond the photocell,
e. g. changing to a faster film or changing
the projection distance. Thus the objective,
condenser position or illuminator may be
changed without affecting k if the cell is
used as described above.
Example: With one particular arrange-
ment of photocell and film,
the meter reading is found to
be 40. A series of photographs
are taken at 1/2, 1/5, 1/10,
1/25 and 1/50 seconds. The
photomicrograph taken at 1/5
second is judged to be the best;
hence k is calculated as follows:
k = meter reading X exposure
time = 40X 1/5 = 8.
Assume now that a new picture
is to be taken at another
magnification (but with the
same film and projection
distance) and that the new
meter reading is 16, therefore
exposure time = k/meter
reading = 8/16 = 1/2 second.
V MICROMETRY
A Particle Size Determination
Linear distances and areas can be
measured with the microscope. This
permits determination of particle size
and quantitative analysis of physical
mixtures. The usual unit of length for
microscopical measurements is the micron
(1 X 10-3mm or about 4 X 10-5mch).
Measuring particles in electron microscopy
requires an even smaller unit, the milli-
micron (1 X 10~3 micron or 10 Angstrom
units). Table 6 shows the approximate
average'size of a few common airborne
materials.
Table 6. APPROXIMATE PARTICLE SIZE OF
SEVERAL COMMON PARTICULATES
Ragweed pollen
Fog droplets
Power plant flyash
(after precipitators)
Tobacco smoke
Foundry fumes
25 microns
20 microns
2-5 microns
0. 2 micron
(200 millimicrons)
0. 1 - 1 micron
(100-1000 millimicrons)
The practical lower limit of accurate
particle size measurement with the light
microscope is about 0. 5 micron. The
measurement of a particle smaller than
this with the light microscope leads to
errors which, under the best circum-
stances, increase to about + 100 percent
(usually +).
One of the principal uses of high resolving
power is in the precise measurement of
I 3-20
-------
Optics and the Microscope
particle size. There are, however, a
variety of approximate and useful proce-
dures as well.
1 Methods of particle size measurement
a Knowing the magnification of the
microscope (product of the magni-
fication of objective and eyepiece).
the size of particles can be esti-
mated. For example, with a 10X
eyepiece and a 16-mm (or 10X)
objective, the total magnification
is 100X. A particle that appears to
be 10-mm at 10 inches from the eye
has an actual size of 10 mm divided
by 100 or 0. 10 mm or 100 microns.
This is in no sense an accurate
method, but it does permit quick
estimation of particle size, the error
in this estimation is usually 10-25
percent.
b Another approximate method is also
based on the use of known data. If
we know approximately the diameter
of the microscope field, we can
estimate the percentage of the
diameter occupied by the object to
be measured and calculate from
these figures the approximate size
of the object. The size of the micro-
scope field depends on both the objec-
tive and the ocular although the latter
is a minor influence. The size of
the field should be determined with
a millimeter scale for each objective
and ocular. If this is done, esti-
mation of sizes by comparison with
the entire field diameter can be quite
accurate (5-10%).
c The movement of a graduated mechan-
ical stage can also be used for rough
measurement of diameters of large
particles. Stages are usually gradu-
ated (with vernier) to read to 0. 1
millimeter, or 100 microns. In
practice, the leading edge of the
particle is brought to one of the lines
of the cross hair in the eyepiece and
a reading is taken of the stage position.
Then the particle is moved across the
field by moving the mechanical stage
in an appropriate direction until the
second trailing edge just touches the
cross-hair line. A second reading is
taken and the difference in the two
readings is the distance moved or the
size of the particle. This method is
especially useful when the particle
is larger than the field, or when the
optics give a distorted image near the
edge of the field.
d The above method can be extended to
projection or photography. The image
of the particles can be projected on a
screen with a suitable light source or
they may be photographed. The final
magnification, M, on the projection
surface (or film plane) is given approxi-
mately by
M = DXO. M. X E. M. /25
where O. M. = objective magnification
E. M. = eyepiece magnification
D = projection distance
from the eyepiece in
centimeters.
The image detail can then be measured
in centimeters and the actual size com-
puted by dividing by M. This method
is usually accurate to within 2-5 percent
depending on the size range of the detail
measured.
e The stated magnifications and/or focal
lengths of the microscope optics are
nominal and vary a bit from objective
to objective or eyepiece to eyepiece.
To obtain accurate measurements, a
stage micrometer is used to calibrate
each combination of eyepiece and
objective. The stage micrometer is
a glass microscope slide that has.
accurately engraved in the center, a
scale, usually 2 millimeters long.
divided into 200 parts, each part repre-
senting 0. 01 millimeter. Thus when
this scale is observed, projected or
photographed, the exact image magni-
fication can be determined. For
example, if 5 spaces of the stage micro-
meter measure 6 millimeters when
projected, the actual magnification is
I 3-21
-------
Optics and the Microscope
5 (0.01)
= 120 times.
This magnification figure can be
used to improve the accuracy of
method 4 above.
f The simplest procedure and the most
accurate is based on the use of a
micrometer eyepiece. Since the
eyepiece magnifies a real image
from the objective, it is possible
to place a transparent scale in the
same plane as the image from the
objective and thus have a scale
superimposed over the image. This
is done by first placing an eyepiece
micrometer scale disc in the eyepiece.
The eyepiece micrometer has an
-arbitrary scale and must be cali-
brated with each objective used. The
simplest way to do this is to place
the stage micrometer on the stage
and note a convenient whole number
of eyepiece micrometer divisions.
The value in microns for each eye-
piece micrometer division is then
easily computed. When the stage
micrometer is removed and replaced
by the specimen, the superimposed
eyepiece scale can be used for accu-
rate measurement of any feature in
the specimen by direct observation,
photography or projection.
2 Calibration of eyepiece micrometer
Each micrometer stage scale has
divisions lOOti (0. 1 mm) apart; one
or two of these are usually subdivided
into 10n (0. 01-mm) divisions. These
form the standard against which the
arbitrary divisions in the micrometer
eyepiece are to be calibrated. Each
objective must be calibrated separately
by noting the correspondence between
the stage scale and the eyepiece scale.
Starting with the lowest power objective
focus on the stage scale, arrange the
two scales parallel and in good focus.
It should be possible to determine the
number of eyepiece divisions exactly
equal to some whole number of
divisions of the stage scale, a distance
readily expressed in microns.
The calibration consists, then, of
calculating the number, of micrpns p^er
eyepiece scale division. To make the
"comparison as accurate as possible, a
large part of each scale must be used
(see Figure 17). Let's assume that
with the low power 16-mm objective
6 large divisions of the s.tage scale
(s. m. d.) are equal to 38 divisions of
the eyepiece scale. This means that
38 eyepiece micrometer divisions (e.m. d.)
are equivalent to 600 microns. Hence:
1 e. m. d. = 600/38
= 15. 8^!
Figure 17
COMPARISON OF STAGE MICROMETER
SCALE WITH EYEPIECE MICROMETER SCALE
Thus when that micrometer eyepiece
is used with that 16-mm objective each
division of the eyepiece scale is equivalent
to 15. 8n, and it'can be used to make an
accurate measurement of any object on
the' microscope stage. A particle, for
example, observed with the 16-mm objec-
tive and measuring 8. 5 divisions on the
eyepiece scale is 8. 5 (15. 8) or 135n in
diameter.
Each objective on your microscope must
be calibrated in this manner.
A convenient way to record the necessary
data and to calculate ^emd is by means
of a table.
I 3-22
-------
Optics and the Microscope
Objective
32-mm
16- mm
4-mm
No.
no.
18
6
1
Table
smd =
emd
= 44
= 38
= 30
7
no.
1800
600
100
emd
= 44
= 38
= 30
1 emd
40.
15.
3.
9 LI
OLJL
33^
Determination of particle size
distribution
The measurement of particle size can
vary in complexity depending on parti-
cle shape. The size of a sphere may be
denoted by its diameter. The size of a
cube may be expressed by the length of
an edge or diagonal. Beyond these two
configurations, the particle "size" must
include information about the shape of
the particle in question, and the
expression of this shape takes a more
complicated form.
Martin's diameter is the simplest means
of measuring and expressing the dia-
meters of irregular particles and is
sufficiently accurate when averaged for
a large number of particles. In this
method, the horizontal or east-west
dimension of each particle which divides
the projected area into halves is taken as
Martin's diameter (Figure 18).
h-P H
Figure 18
MARTIN'S DIAMETER
The more particles counted, the more
accurate will be the average particle
size. Platelike and needlelike particles
should have a correction factor applied
to account for the third dimension since
all such particles are restricted in their
orientation on the microscope slide.
When particle size is reported, the
general shape of the particles as well as
the method used to determine the
"diameter" should be noted.
Particle size distribution is determined
routinely by moving a preparation of
particles past an eyepiece micrometer
scale in such a way that their Martin's
diameter can be tallied. All particles
whose centers fall within two fixed
divisions on the scale are tallied. Move-
ment of the preparation is usually
accomplished by means of a mechanical
stage but may be carried out by rotation
of an off-center rotating stage. A sample
tabulation appears in Table 8. The eye-
piece and objective are chosen so that
at least six, but not more than twelve,
size classes are required and sufficient
particles are counted to give a smooth
curve. The actual number tallied (200 -
2, 000) depends on particle shape
regularity and the range of sizes. The
size tallied for each particle is that
number of eyepiece micrometer divisions
most closely approximating Martin's
diameter for that particle.
4 Calculation of size averages
The size data may be treated in a variety
of ways, one simple, straightforward
treatment is shown in Table 9. For a
more complete discussion of the treat-
ment of particle size data see Chamot
and Mason's Handbook of Chemical
Microscopy(3)t page 26.
The averages^ with respect to number,
dj, surface, d3, and weight or volume,
£4, are calculated as follows for the
data in Table 9.
I 3-23
-------
Optics and the Microscope
Table 8. PARTICLE SIZE TALLY FOR A SAMPLE OF STARCH GRAINS
ins
(emd*)
Number of particles
Total
rwj rt-w n-*4 rt-*j rt-u r-su
rt-4j rtsj MHJ rr-w r*-u rt-*j
«-4j M-W I-MJ rt-ij r*-*j 111
rt-*j
rt-ia
tt-u r-*-*j r»-*j
t-t-w
rt-u rt-w rt-*-i rtHj 11-4.1 r-*oa
rt-*j
1 1
rt-u
16
98
110
107
71
ri-*j
1 1
45
II
3
470
emd exeniece mlcrometer-dixiaiona
d1 = Znd/Zn = 1758/470
= 3. 74 emd X 2. 82* = 10. 5(i
d3 = Znd3/ 2nd2 = 37440/7662
= 4. 89 emdx 2.82 = 13. Sp.
d4 = Snd4/Znd3 = 199194/37440
= 5.32 emdX 2. 82 = 15.0^
*2. 82 microns per emd
(determined by calibration of the
eyepiece-objective combination
used for the determination).
Cumulative percents by number,
surface and weight (or volume) may be
plotted from the data in Table 9. The
calculated percentages, e. g.
d = 15
nd4 X 100
d = 1
d = 1
for the cumulative weight or volume
curve, are plotted against d. Finally,
the specific surface, Sm, in square
meters per gram, m, may be calculated
if the density, O, is known, the surface
average d3, is used.
IfD=l.l, Sm = 6/daD = 6/13. 8(1. 1)
= 0. 395m2/g.
I 3-24
-------
Optics and the Microscope
Table 9. CALCULATIONS FOR PARTICLE SIZE AVERAGE
d
(Aver. diam.
in emd)
1
2
3
4
5
6
7
8
n
16
98
110
107
71
45
21
2
nd
16
196
330
428
355
270
147
16
nd2
16
392
990
1712
1775
1620
1029
128
nd3
16
784
2970
6848
8875
9720
7203
1024
nd4
16
1568
8910
27392
44375
58320
50421
8192
470 1758 7662 37440 199194
B Counting Analysis
Mixtures of particulates can often be
quantitatively analyzed by counting the
total number of particulates from each
component in a representative sample.
The calculations are, however, compli-
cated by three factors: average particle
size, particle shape and the density
of the components. If all of the compon-
ents were equivalent in particle size,
shape and density then the weight per-
centage would be identical to the number
percentage. Usually, however, it is
necessary to determine correction factors
to account for the differences.
When properly applied, this method can
be accurate to within + 1 percent and,
in special cases, even better. It is often
applied to the analysis of fiber mixtures
and is then usually called a dot-count
because the tally of fibers is kept as the
preparation is moved past a point or dot
in the eyepiece.
A variety of methods can be used to
simplify recognition of the different
components. These include chemical
stains or dyes and enhancement of optical
differences such as refractive indices,
dispersion or color. Often, however, one
relies on the differences in morphology.
e. g. counting the percent of rayon fibers
ln~a~sarm)le of "silk".
Example 1: A dot-count of a mixture of
fiberglass and nylon shows
nylon
fiberglass
262
168
Therefore
% nylon = 262/(262 + 168)X 100
= 60. 9% by number.
However, although both fibers are smooth
cylinders, they do have different densities
and usually different diameters. To
correct for diameter one must measure
the average diameter of each type of fiber
and calculate the volume of a unit length
of each.
aver. diam. volume of
p. 1-ji slice, ji3
nylon
fiberglass
18. 5
13.2
268
117
The percent by volume is, then:
262X268
_
(262X268K(168X117)
= 78. 1% by volume.
Still we must take into account the density of
each in order to calculate the weight percent.
I 3-25
-------
Optics and the Microscope
If the densities are 1. 6 for nylon and 2. 2
for glass then the percent by weight is
262 X 2C8 X 1. C
% nylon = (262 x 2fig x i.G)+(if,8X 117 X 2.2)
= 72% by weight.
Example 2 A count of quartz and
gypsum shows
quartz
gypsum
283
467
To calculate the percent by weight we must
take into account the average particle size,
the shape and the density of each.
The average particle size with respect to
weight, d4, must be measured for each
and the shape factor must be determined.
Since gypsum is more platelike than quartz
each particle of gypsum is thinner. The
shape factor can be approximated or can be
roughly calculated by measuring the actual
thickness of a number of particles. We
might find, for example, that gypsum parti-
cles average 80% of the volume of the aver-
age quartz particle, this is our shape factor.
The final equation for the weight percent is:
quartz =
283 X nd4/6X Dq
X 100
238 X ird4/6X Dq + 467 X TT d|/6 X 0. 80 X Dg
X 100
where Dq and Dg are the densities of quartz
and gypsum_respe£tively, 0. 80 is the shape
factor and d4 and d4 are the average parti-
cle sizes with respect to weight for quartz
and gypsum respectively.
ACKNOWLEDGMENT: This outline was
prepared by the U. S. Public Health Service,
Department of Health, Education and Welfare,
for use in its Training Program.
REFERENCES
1 Bunn, C.W. Crystal Growth from Solution.
Discussions of the Faraday Society No. 5.
132. Gunery and Jackson. London. (1949),
2 Loveland, R.P., J. Roy. Micros. Soc.
79, 59. (1960).
3 Chamot, Emile Monnin, and Mason,
Clyde Walter. Handbook of Chemical
Microscopy, Vol. 1, third ed. John
Wiley and Sons, New York (1959).
I 3-26
-------
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.
Water quality management, including
pollution control, thus looks to all
branches of aquatic science in efforts
to coordinate and improve man's
relationship with his aquatic environment.
II SOME FACTS ABOUT WATER
A Water is the only abundant liquid on our
planet. It has many properties most
unusual for liquids, upon which depend
most of the familiar aspects of the world
about us as 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
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 (4°C)
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"
BI.21b.2.70
I 4-1
-------
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
Figure 1
I 4-2
-------
The Aquatic Environment
TABLE 2
EFFECTS OF TEMPERATURE ON DENSITY
OF PURE WATER AND ICE*
Temperature (°C)
Density
Water
Ice **
-10
- 8
- 6
- 4
- 2
0
2
4
6
8
10
.99815
.99869
.99912
.99945
.99970
.99987
.99997
1.00000
.99997
.99988
0.00973
.9397
.9360
.9020
.9277
.9229
.9168
* 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 m, etc.), having densities
at 1 atm. pressure ranging from 1.1595
to 1. 67. These are of extremely restricted
occurrence and may be ignored in 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 DO C. It is adhesive and
may build up on submerged objects
as "anchor ice", but it is still
typical ice.
1) Seasonal increase in 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
thermocline or shear-plane.
2) While for certain theoretical
purposes a thermocline 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 thermocline.
3) Obviously the greater the
temperature differences
between epilimnion and
hypolimnion and the sharper
the gradient in the thermocline.
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
Oo 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
chemical and physical
characteristics. Such periods
are called overturns and
I 4-3
-------
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 Olg tends to go off
first in the process of
evaporation, leading to the
relative enrichment of air by
Oig and the enrichment of
water by O17 and Olg. This
can lead to a measurably
higher Ojo content in warmer
climates. Also, the temper-
ature of water in past geologic
ages can be closely estimated
from the ratio of O]^ in the
carbonate of mollusc shells.
b Dissolved and/or suspended solids
may also affect the density of
natural waters.
TABLE 3
EFFECTS OF DISSOLVED SOLIDS
ON DENSITY
Dissolved Solids
(Grams per liter)
0
1
2
3
10
Density
(at 40C)
1.00000
1.00085
>
1.00169
1.00251
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.
The viscosity of water is greater at
lower temperatures (see Table 4).
This is important not only in 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.
I 4-4
-------
The Aquatic Environment
TABLE 4
VISCOSITY OF WATER (In millipoises at 1 atm)
Temp, o C
-10
- 5
0
5
10
30
100
Dissolved solids in g/L
0
26.0
O1 A
21.4
17.94
15.19
13.10
8.00
2.84
5
18.1
15.3
13.2
8.1
10
18.24
15.5
13.4
8.2
30
18.7
16.0
13.8
8.6
Surface tension has been reported
from 34 to 160 atm. This has
biological as well as physical sig-
nificance. Organisms whose body
surfaces cannot be wet by water can
either ride on the surface film, or in
some instances may be "trapped" on
the surface film and be unable to
re-enter the water.
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-
ically, 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.
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 hypolimnion. 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.
I 4-5
-------
The Aquatic Environment
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 cur rents )are
arhythmic water movements which
have had major study only in
oceanography. 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 estuaries 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.
6 The pH of pure water has been deterr
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.
I 4-6
-------
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.
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 CO, affects the heat transmission of
the atmosphere.
C Organisms respond to and in turn affect
their environment. Man is one of the
most influential.
ID ECOLOGY IS THE STUDY OF THE
INTERRELATIONSHIPS BETWEEN
ORGANISMS, AND BETWEEN
ORGANISMS 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 pacts (Odum, 1959).
1 From a functional standpoint, an
ecosystem has two component parts.
(Figures 1. 2,3)
a Autotrophic (self-nourishing)
organisms are able to fix light
energy and manufacture food from
simple inorganic substances.
b Heterotrophic (other-nourishing)
organisms utilize, rearrange,
and decompose the complex
materials synthesized by the
autotrophs.
2 From a structural standpoint, it is
convenient to recognize four con-
stituents as comprising an ecosystem.
a Abiotic substances are basic or
essential mineral elements and
compounds.
b Autotrophic (holophytic) organisms
are the producers, largely the
green plants.
c Heterotrophic (or holozoic)
organisms are chiefly animals
that ingest or consume other
organisms or particulate organic
matter.
BI.21b.2.70
I 4-7
-------
i
oo
ESSENTIAL COMPONENTS
NON-ESSENTIAL
COMPONENTS
(SEWAGEJ
PRIMARY
DECOMPOSERS
HERBIVORES
CARNIVORES
SCAVENGERS
SECONDARY
DECOMPOSERS
TRANSFORMERS
f MINERALIZED
( EFFLUENT )(oETRITUS
SUBSTRATE
PRODUCERS
CONSUMERS
TERMINAL
r NON-LIVING
COMPONENTS
LIVING
COMPONENTS
PRINCIPAL STEPS AND COMPONENTS IN THE TRICKLING FILTER ECOSYSTEM
FROM. COOKE. ECOLOGY. 40(2) 1959 BI. ECO pi. 1 7 59
CD
a
I
H"
O
!
§
3
(D
-------
Aquatic Environment
Rg«r« 2. 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; IIMA, primary consumers
(herbivores)-bottom forms; IIMB, primary consumers (herbivores )-zooplankton; III-2, secondary consumers (car-
wvorei); III-3. tertiary consumers (secondary carnivores); IV, decomposers-bacteria and fungi of decay.
d Decomposers are heterotrophic
organisms, chiefly bacteria and
fungi that return or reduce the
complex compounds of dead
protoplasm to their original
mineral condition.
B Functioning of the Ecosystem
1 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 inter-
connected.
2 A food web is the interlocking pattern
of food chains in an ecosystem.
(Figure 1) In complex natural com-
munities, organisms whose food is
obtained by the same number of steps
are said to belong to the same trophic
(feeding) level.
3 Trophic levels
a First - Green plants (producers)
(Figure 4)fix biochemical energy and
synthesize basic organic substances.
b Second - Plant eating animals
(herbivores) depend on the
producer organisms for food.
c Third - Primary carnivores,
animals which feed on herbivores.
d Fourth - Secondary carnivores
feed on primary carnivores.
e Last - Ultimate carnivores are
the last or ultimate level of
consumers.
4 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.
5 Trophic structure of the ecosystem
The interaction of the food chain
phenomena (with energy loss at each
transfer) results in various com-
munities having definite trophic
I 4-9
-------
The Aquatic Environment
^/T\ Light'.':;;.
- j" * ^^ *" i'" % '.^j»- w->-^-^ T L i "i^'ai '' ~ i tm r ' \^L^p^r^^*«^^7 -^^^
Nutrient
supply
Bacterial
action
Death and decay
\
Predatory
animals
)TE ROT HO Ills
CAiOilVORKS)
Figure 3. A MARINE ECOSYSTEM (After Clark, 1954 and Patten, 1966)
I 4-10
-------
The Aquatic Environment
1
\r<
(a)
Decomposers 11 Carnivores (Secondar
[| Carnivores (Primary
1 | Herbivores
Producers |
(b)
A
1 1
1 I
(c)
,, &O -
f
-------
The Aquatic Environment
REFERENCES 5 Odum, E.P. Fundamentals of Ecology.
W.B. Saunders Company,
1 Clarke, G. L. Elements of Ecology. Philadelphia and London. 1959.
John Wiley & Sons, New York. 1954.
6 Patten, B.C. Systems Ecology.
2 Cooke, W.B. Trickling Filter Ecology. Bio-Science. 16(9). 196.6.
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.
I 4-12
-------
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 ASA
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.21b.2.70
I 4-13
-------
The Aquatic Environment
6 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
,ane 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 Material
Sand
Marl
Fine Gravel
Gravel and silt
Coarse gravel
Moss on fine gravel
Fissidens (moss) on coarse gravel
Ranunculus (water buttercup)
Watercress
Anacharis (water weed)
Relative Productivity
1
6
9
14
32
89
111
194
301
452
'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.
I 4-14
-------
The Aquatic Environment
1 Youthful streams, especially on rock
or sand substrates are low in essential
nutrients. Temperatures in moun-
1 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
Sand
Pebbles
Clay
Flat rubble
Block rubble
Shelving rock
Relative Productivity
1
4
8
9
11
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.
) 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.
I 4-15
-------
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.
Reservoir discharges also profoundly
affect the DO, temperature, and
turbidity in the stream below a dam.
Too much fluctuation in 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 Resei voirs 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. it-xix, 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 Ruttner, Franz.
Limnology.
Press, pp.
Fundamentals of
University of Toronto
1-242. 1953.
Tarzwell, Clarence M. Experimental
Evidence on the Value of Trout 1937
Stream Improvement in Michigan.
American Fisheries Society Trans.
66:177-187. 1936.
I 4-16
-------
The Aquatic Environment
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 Sanitary Engineering Center.
Cincinnati, Ohio. No. SEC TR W61-3.
7 Ward and Whipple. Fresh Water
Biology. (Introduction). John
Wiley Company. 1918.
This outline was prepared by H. W. Jackson,
Chief Biologist, National Training Center.
FWPCA. Cincinnati, OH 45226.
I 4-17
-------
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)
Flgura 1. IDE WATBt C1CU
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 i
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
Na
K
Ca
Mg
Cl
so4
C03
Delaware River
at
Lambertville, N.J.
6.70
1.46
17.49
4.81
4.23
17 49
32.95
Rio Grande
at
Laredo, Texas
14.78
.85
13.73
3.03
21.65
30.10
11.55
Sea Water
30.4
1. 1
1.16
3.7
55.2
7.7
»-Hco3 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
BI.21b.2.70
I 4-19
-------
The Aquatic Environment
Type of environment
and general direction
of water movement
Salinity
Degree of instability
Temperature
Water
elevation
Vertical
strati-
fication
Avail -
ability
of
nutrients
(degree)
Turbidity
Riverine
Oceanic
Figure 2 . 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.
Neritic - Relatively shallow-water
zone which extends from the high-
tide mark to the edge of the
continental shelf. (Figure 3)
I 4-20
-------
The Aquatic Environment
Pelagiat
-J600
Primary 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.
Euphotic zone - Waters into which
sunlight penetrates (often to the
bottom in the neritic zone). The
zone of basic productivity. Often
extends to 600 feet below the
surface.
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.
1) Physical factors more con-
stant than in bathyal zone.
2) Producers absent and
consumers not as abundant
as in the bathyal zone.
I 4-21
-------
The Aquatic Environment
IE 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 giUs.
IV FACTORS AFFECTING THE DISTRI-
BUTION OF MARINE ORGANISMS
A 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 in 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
Fresh Water
Stenohaline
Marine
Stenohaline
Salinity
ca. 35
Figure 4. Salinity Tolerance of Organisms
b Eurythermal - wide tolerance to
temperature changes.
I 4-22
-------
The Aquatic Environment
SNAILS
Littorina neritoides
L. rudis
L. obtusata
L. littorea
BARNACLES
Chthamalus stellatus
Balanus balanoides
B. perforatus
%&?%&*£* ^° * «4%T>
l^mm»Q
\ \f\il J 1 -I HI i V»v> IM\\O^I.C
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)
I 4-23
-------
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 in 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. L 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 Publs.
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,
FWPCA. Cincinnati. OH 45226.
I 4-24
-------
CHAPTER II
IDENTIFICATION OF PLANKTON AND ASSOCIATED ORGANISMS
Structure and Function of Cells 1
Aquatic Organisms of Significance in Plankton Surveys 2
Types of Algae 3
Blue-Green Algae 4
Green and Other Pigmented Flagellates 5
Filamentous Green Algae 6
Coccoid Green Algae 7
Diatoms 8
Filamentous Bacteria 9
Protozoa, Nematodes, and Rotifers 10
Free-Living Amoebae and Nematodes 11
Animal Plankton 12
-------
STRUCTURE AND FUNCTION OF CELLS
I INTRODUCTION
What are cells? Cells may be defined as the
basic structural units of life. The cell has
many different parts which carry on the
various functions of cell life. These are
called organelles ("little organs").
A The branch of biology which deals with the
form and structure of plants and animals
is called "Morphology. " The study of the
arrangement of their several parts is
called "anatomy", and the study of cells
is called "cytology".
B There is no "typical" cell, for cells differ
from each other in detail, and these
differences are in part responsible for the
variety of life that exists on the earth.
E FUNDAMENTALS OF CELL STRUCTURE
A How do we recognize a structure as a cell?
We must look for certain characteristics
and/or structures which have been found
to occur in cells. The cell is composed
of a variety of substances and structures,
some of which result from cellular
activities. These include both living and
non-living materials.
1 Non-living components include:
a A "cell wall" composed of cellulose
may be found as the outermost
covering of many plant cells.
b "Vacuoles" are chambers in the
protoplasm which contain fluids of
different densities (i.e., different
from the surrounding protoplasm).
2 The "living" parts of the cell are called
"protoplasm. " The following structures
are included:
a A thin "cell membrane" is located
just inside the cell wall. This
membrane may be thought of as the
outermost layer of protoplasm.
b In plant cells the most conspicuous
protoplasmic structures are the
"chloroplasts", which contain
highly organized membrane systems
bearing the photosynthetic pigments
(chlorophylls, carotenoids, and
xanthophylls) and enzymes.
c The "nucleus" is a spherical body
which regulates cell function by
controlling enzyme synthesis.
d "Granules" are structures of small
size and may be "living" or
non-living" material.
e "Flagella" are whip-like structures
found in both plant and animal cells.
The flagella are used for locomotion,
or to circulate the surrounding
medium.
f "Cilia"resemble short flagella, found
almost exclusively on animal cells.
In the lower animals, cilia are used
for locomotion and food gathering.
g The "pseudopod", or false foot, is
an extension of the protoplasm of
certain protozoa, in which the
colloidal state of the protoplasm
alternates from a "sol" to a "jel"
condition from time to time to
facilitate cell movement.
h "Ribosomes" are protoplasmic bodies
which are the site of protein
synthesis. They are too small
(150 A in diameter)to be seen with
a light microscope.
i "Mitochondria" are small mem-
branous structures containing
enzymes that oxidize food to produce
energy transfer compounds (ATP).
BI.CEL. la.3.70
H 1-1
-------
Structure and Function of Cells
B How basic structure is expressed in some
major types of organisms.
We can better visualize the variety of cell
structure by considering several specific
ceUs.
1 Bacteria have few organelles, and are
so minute that under the light
microscope only general morphological
types (i.e., the three basic shapes;
rods, spheres, and spirals) can be
recognized. The following structures
have been defined:
a The "capsule" is a thick protective
covering of the cell exterior, con-
sisting of polysaccharide or
polyp eptide.
b The cell wall and plasma membrane
are present.
c Although no well defined nucleus is
visible in bacterial cells, the
electron microscope has revealed
areas of deoxyribose nucleic acid
(ONA) concentration. This sub-
stance is present within the nucleus of
of higher cells, and is the genetic
or hereditary material.
d Some types of bacteria contain a
special type of chlorophyll
(bacteriochlorophyll) and carry on
photosynthesis.
2 The blue-green algae are similar to the
bacteria in structure, but contain the
photosynthetic pigment chlorophyll a.
a Like the bacteria, these forms also
lack an organized nucleus (the
nuclear region is not bounded by a
membrane).
b The chlorophyll-bearing membranes
are not localized in distinct bodies
(chloroplasts), but are dispersed
throughout the cell.
c Gas-filled structures called
"pseudovacuoles" are found in some
types of blue-greens.
The green algae as a group include a
great variety of structural types,
ranging from single-celled non-motile
forms to large motile colonies. Some
types are large enough to resemble
higher aquatic plants.
a The chloroplasts are modified into
a variety of shapes and are located
in different positions. Examples
of chloroplast shape and position are:
1) Parietal - located on the
periphery of the cell; usually
cup-shaped and may extend
completely around the inner
surface of the plasma membrane.
2) Discoid - also located on the
periphery of the cell, but are
plate-shaped; usually many per
ceU.
3) Axial - lying in the central axis
of the cell; may be ribbon-like
or star-shaped.
4) Radial - have arms or processes
that extend outward from the
center of the cell (radiate),
reaching the plasma membrane.
5) Reticulate - a mesh-like network
that extends throughout volume
of the cell.
b Located in the chloroplasts may be
dense, proteinaceous, starch-
forming bodies called "pyrenoids".
The flagellated algae possess one-to-
eight flagella per cell. The chloro-
plasts may contain brown and/or red
pigments in addition to chlorophyll.
a Reserve food may be stored as
starch (Chlamydomonas) paramylon
(Euglena), or as oil.
The protozoa are single-celled
animals which exhibit a variety of
cell structure.
H 1-2
-------
Structure and function of Cells
The amoebae move by means of
pseudopodia, as described
previously.
The flagellated protozoa
(Mastigophora) possess one or more
flagella.
The ciliates are the most highly
modified protozoans. The cilia may
be more or less evenly distributed
over the entire surface of the cell,
or may be localized.
IH FUNCTIONS OF CELLS
What are the functions of cells and their
structural components? Cellular function
is called "life", and life is difficult to define.
Life is characterized by processes commonly
referred to as reproduction, growth, photo-
synthesis, etc.
A Microorganisms living in surface waters
are subjected to constant fluctuations in
the physical and chemical characteristic
of the environment, and must constantly
modify their activities.
1 The cell requires a cource of chemical
energy to carry on life processes and
successfully compete with other
organisms. Plant cells may obtain
this energy from light, which is
absorbed by chlorophyll and converted
into ATP or food reserves, or from
the oxidation of food stuffs. Animal
cells obtain energy only from the
oxidation of food.
2 Cells must obtain raw materials from
the environment in order to grow and
carry out other life functions. Inorganic
and organic materials may be taken up
by passive diffusion or by "active
transport". In the later process,
energy is used to build up and maintain
a higher concentration of a substance
(such as phosphate) inside the cell than
is found outside. Algae are able to
synthesis organic matter from inorganic
raw materials (carbon dioxide and
water), with the aid of energy derived
from light, whereas animal cells must
obtain their organic matter "ready-
made" by consuming other organisms,
organic debris, or dissolved organics.
IV SUMMARY
The cell is made up of many highly special-
ized substructures. The types of sub-
structures present, and their appearance
(shape, color, etc,) are very important in
understanding the role of the organism in
the aquatic community, and in classification.
REFERENCES
1 Bold, H. C. Cytology of algae. In: G.M.
Smith, (ed.), Manual of Phycology.
Ronald Press. 1951.
Bourne, GeoffryH.,
Cell Physiology.
Press. 1964.
ed. Cytology and
3rd ed. Academic
3 Brachet, Jean. The Living Cell.
Scientific American. 205(3). 1961.
4 Corliss, John O. Ciliated Protozoa.
Pegamon. 1961.
5 Fritsch, F. E. The structure and
reproduction of the algae. Cambridge
Univ. Press. 1965.
6 Frobisher, M. Fundamentals of
microbiology. 7th edition. W.B.
Saunders Co., Philadelphia. 1962.
7 Round, F.E. The biology of the algae.
St. Martin's Press. New York. 1965.
This outline originally prepared by Michael
E. Bender, Biologist, formerly with
Training Activities, FWPCA, SEC. and
revised by Cornelius I. Weber, March 1970.
II 1-3
-------
AQUATIC ORGANISMS OF SIGNIFICANCE IN PLANKTON SURVEYS
I INTRODUCTION
A Any organism encountered in a survey is
of significance. Out problem is thus not
to determine which are of significance but
rather to decide, "what is the significance
of each?"
B The first step in interpretation is
recognition.
C Recognition implies identification and an
understanding of general relationships.
The following outline will thus review the
general relationships of living (as con-
trasted to fossil) organisms, and briefly
describe the various types.
H THE GENERAL RELATIONSHIPS OF
LIVING ORGANISMS
A Living organisms have been long 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 traditional "plant" and
"animal. " The accompanying chart con-
sequently shows the fungi as a third
"kingdom."
B The three groups are essentially defined
as follows on the basis of their nutritional
mechanisms:
1 Plante: photosynthetic, synthetizing
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 metabolized and
reduced to the mineral condition.
Ecologically known as REDUCERS.
Each of these groups includes simple,
single-celled representatives, per-
sisting at lower levels on the evolutionary
stems of the higher organisms (as well
as the higher and more complex types).
1 These groups span the gaps between
the higher kingdoms with a multitude
of transitional form. They are
collectively called the PROTISTA.
2 Within the Protista, two principle
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.
Distributed throughout these groups will
be found 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.
2 Submersed types typically do not
extend to the surface.
BI.AQ. 10d.4.70
II 2-1
-------
Aquatic Organisms of Significance in Plankton Surveys
CONSUMERS
PRO DUCERS
REDUCERS
NUTRIENT
MINERALS
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
organisms, may be green, red, brown,
etc.
"Diatoms" have "pillbox" structure of
SiOp - may move. Extremely common.
Many minute in size, but colonial forms
may produce hair-like filaments.
Golden brown in color.
"Non-motile green algae" lack
locomotor 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.
II 2-2
-------
Aquatic Organisms of Significance in Plankton Surveys
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.
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.
3 CNIDARIA (=COELENTERATA) include
corals, marine and freshwater jelly
fishes, marine and freshwater hydroids.
4 PLATYHELMINTHES are the flat worms
such as tape worms, flukes and Planaria.
NEMATHELMINTHES are the round
worms and include both free-living
forms and many dangerous parasites.
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 BRACMOPODS are bivalved marine
organisms usually observed as fossils.
10 ANNELIDS are the segmented worms
such as earthworms, sludge worms
and many marine species.
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
appendages and a chitinous exoskelton.
a CRUSTACEA are divided into a
cephalothorax and abdomen, and
have many pairs of appendages,
including two pairs of 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 tem-
porary pools.
3) COPEPODS are marine and
freshwater microcrustacea--
swim by means of unbranched
antennae.
4) OSTRA CODS are the microscopic
"clams with legs. " Generally
substrate oriented.
5) ISOPODS are dorsoventrally
compressed; called sowbugs.
Terrestrial and aquatic, marine
and freshwater.
H 2-3
-------
Aquatic Organisms of Significance.in Plankton Surveys
6) AMPfflPODA - known as scuds.
laterally compressed. Marine
and freshwater.
7) DECAPODA - crabs, shrimp,
crayfish, lobsters, etc. Marine
and freshwater.
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 fresh-
water moths
ARACHINIDA - body divided into
cephalothorax and abdomen; 4 pairs
of legs - spiders, scorpions, ticks
and mites. Few aquatic represent-
atives except for the freshwater
mites and tardigrades.
C CHORDATA
1 PROCHORDATES - 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 - snails, lizards and
turtles
d MAMMALS - whales and other
warm-blooded vertebrates with hair
e AVES - birds - warm-blooded
vertebrates with feathers
VI THE CLASSIFICATION OF ORGANISMS
A INTRODUCTION
There are few major groups of orga-
nisms that are either exclusively
terrestrial or generally aquatic. The
following remarks apply to both, however,
primary attention will be directed to
aquatic types.
B 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 under-
stood by anyone working with organisms
however.
H 2-4
-------
Aquatic Organisms of Significance in Plankton Surveys
RELATIONSHIPS BETWEEN FREE LIVING AQUATIC ORGANISMS
Energy Flows from Left to Right, General Evolutionary Sequence is Upward
PRODUCERS
Organic Material Produced,'
Usually by Photosynthesis "
CONSUMERS
Organic Material Ingested or
Consumed
Digested Internally
REDUCERS
Organic Material Reduced
by Extracellular Digestion
and Intracellular Metabolism
to Mineral Condition
ENERGY STORED
ENERGY RELEASED
ENERGY RELEASED
Flowering Plants and
Gymnosperms
Club Mosses, Ferns
Liverworts, Mosses
Multicellular Green
Algae
Red Algae
Brown Algae
Arachnids
Insects
Crustaceans
Mammals
Birds
Reptiles
Segmented Worms Amphibians
Molluscs Fishes
Bryozoa
Rotifers
Roundworms
Flat worms
Sponges
Primitive
Chordates
Echinoderms
Coelenterates
Basidiomycetes
Fungi Imperfecti
Ascomycetes
Higher Phycomycetes
DEVELOPMENT OF MULTICELLULAR OR COENOCYTIC STRUCTURE
H I G" H E R P R 0 T I S T A
Protozoa
Unicellular Green Algae
Diatoms
Pigmented Flagellates
Amoeboid
Flagellated,
(non-pigmented)
Cilliated
Suctoria
Lower
Phycomycetes
(Chytridiales, et. al )
DEVELOPMENT OF A NUCLEAR MEMBRANE
LOWER PROTISTA
(or M o n -e r a )
Blue Green Algae
Phototropic Racteria
Chemotro|iic Hacteria
Actinomycetes
Spuochaetes
Sapt ophytic
Bacterial
Types
II 2-5
-------
Aquatic Organisms of Significance in Plankton Surveys
Names are the "key number", code
designation", or "file references"
which we must have to find information
about an unknown organism.
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 subdivisions. 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).
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 through-
out 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 Ichthyolo-
gists and Herpetologists.
The system of biological nomenclature
is regulated by international congresses.
a It is based on a system of groups
and super groups, of which the
foundation (which actually exists in
nature) is the species.
b 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.
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).
a The generic (genus) name is always
capitalized and the species name
written with a small letter. They
should also be underlined or printed
in italics when used in a technical
sense. For example:
Homo sapiens - (=sentiens) modern
man
Homo heidelbergensis - heidelberg
man
Homo neanderthalis - neanderthal
man
Oncorhynchus gorbuscha - pink
salmon
Oncorhynchus kisutch - coho salmon
Oncorhynchus tshawytscha -
Chinook salmon
b 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:
II 2-6
-------
Aquatic Organisms of Significance in Plankton Surveys
Gomphonema olivaceum
Gomphonema parvulum
Gomphonema abbreviatum
three distinct species which have
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
Gomphonema olivaceum is:
Kingdom
Phylum
Class
Order
Family
Genus
Species
Plantae
Chrysophyta
Bacillariophyceae
Pennales
Gomphonema ceae
Gomphonema
olivaceum
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.
It should be emphasized that since
most categories above the species
level are essentially human con-
cepts, there is often divergence of
opinion in regard to how certain
organisms should be grouped.
Changes result as knowledge grows.
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.
REFERENCE
Whittaker, R.H.
of Organisms.
1969.
New Concepts of Kingdoms
Science 163:150-160.
a 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 as:
Superorder, Order, Suborder, etc..
Still other category names such as
"tribe", "division", "variety",
"race", "section", etc., are used
on occasion.
This outline was prepared by H. W. Jackson,
Chief Biologist, National Training Center.
Revised 1970 by R. M. Sinclair, Aquatic
Biologist, National Training Center, FWPCA,
Cincinnati, OH 45226.
H 2-7
-------
TYPES OF ALGAE
I INTRODUCTION
A A Igae - Simple plants with an autotrophic
mode of nutrition. They contain chloro-
phyll, but lack the highly differentiated
reproductive and body structures of higher
plants. The algae range in size from less
than one micron, to the giant kelp more
than 300 ft. in length.
B Algae are carried about by water currents,
wind, aquatic animals, water fowl, and
insects, and are found everywhere. They
grow actively at temperatures ranging
from -2°C to 6QOC.
C The algae play a more important role in
aquatic habitats than on land. They serve
as the basic energy source for other
aquatic organisms, providing food for
zoomicrobes, higher invertebrates and
fish.
D There are two basic types of freshwater
algal communities:
1 The phytoplankton - free floating algae
2 Phycoperiphyton - attached or bottom
forms
II Algae are classified by their pigments,
food pigments, food reserves, morphology,
and method of reporduction. Recent classifi-
cation schemes recognize eight major groups
(phyla).
A Blue-green (Cyanophyta)
B Green (Chlorophyta)
C Yellow-brown (Chrysophyta)
D Euglenoid (Euglenophyta)
E Dinoflagellates (Pyrrophyta)
F Red (Rhodophyta)
G Brown (Phaeophyta)
H Chloromonads (Chloromonadophyta)
The most numerous freshwater algae are the
yellow browns, greens, and blue-greens.
Ill BLUE-GREEN ALGAE
This group contains many organisms that
form nuisance blooms in eutrophic waters
in later summer. They are considered to
be the most primitive algae, and are
similar to the bacteria in many respects.
They lack many of the basic cell structures
found in other algae and terrestrial plants,
and have no sexual reproduction or motile
forms. Many have the ability to fix nitrogen.
A The chlorophyll-bearing membranes are
not organized to form distinct bodies
(chloroplasts).
B The cells contain chlorophyll a, but are
blue-green because of the presence of the
blue pigment, phycocyanin (Table 1).
C The nucleus is not enclosed by a
membrane.
D Reproduction is by cell division, colony
fragmentation, and spore formation:
Spores of three types are observed.
1 Akinetes - enlarged, (often) thick-
walled resting cells (see Anabaena).
2 Heterocysts - appear "empty", but
have been observed to germinate
(see Anabaena).
3 Endospores - formed by repeated
division of protoplast in a cell
(Chamaeosiphon).
BI.MIC.cla.22.3.70
II 3-1
-------
"IVpes of Algae
TABLE 1
The distribution of pigmrnts in the algal groups. = major
pigmenl(s) of (he group; 3 = a pigment comprising less than half the
total pigment content; O = present in small amounts. The data given by
earlier authors varies considerably and the present table is compiled from
tables in Fogg (1953), Strain (1958), Goodwin (1960), Egle (1960) and
Haxo and O'Heocha (1960) with additions from their discussions.
X. Alfil
^S^ tmw
\.
Fitments ^N^^
^\
"chlorCh
Chlorophyll!
Chlorophyll <
Chlorophyll 1
Chlorophyll i
Cntnu
a'Carotene
p^Carotenc
r^Caroune
-Carotene
Fuwaccne
Zeaunthin
Vulounlhin
Flavoxaxilhm
Fucounthin
Neofucoximhm (A and B)
Dutounthin
Diadinoxanlhin
Neodiadinonnlhin
Dmoxanlhin
Neodinoxanlhin
Pendimn
Mrmuthint-Echmenone]
Myraunlhophtll
OKillounlhui
Ailuaalhin
Unlnovm nanlhophyU
HUnUM
R-ph»coer)thrm
R'phyooc) an in
C-phi coert ihf in
Alloph) cocyanin
T
o
o
o
0'
^
j
1
8
o
o
o
8
8
a
j
I
»
T
o
o1
o
R
1
4
4
o
o
0
o
o
o
o
"
£
e
o
^
0
o
o
Q
a
j
|
0
*0°
±0
0
±o
o
o
Q
Q
1
I
1
0
o
o
A
3
1
.',
1
0«
V
*
z
§:
0
±o
o
9
8
z
REMARKS
Bui not In all fenerft
Kol in higher planu
In Ktriiflt uriuam
Tlraunlhm'
Occun il chroma-
protein
Alu^^huHr..^
MMU and PMWI/AM
Alia in Cryptomonadi
Also in Cryptoinoiuai.
but tbwrpuon priu
net Ihe lame
In PprpA/ra n*i«Afm
I. 5-4 times Ihe content of/9-nrolene and lutein, but famtles ofFuna almost cntiivl> ^-carotene
B. aomelimes in gnalcr irnouDl than P carotene
S In large amounts possibly in Ihe c>loplism
4 Dominant in some speciei 1 he carolenoid content! of Euitta* iff. are hifher than other ali
-------
Types of Algae
E For convenience, the phylum is divided
into two groups:
1 Coccoid - cells variously shaped,
solitary or in masses, non-filamentous
2 Filamentous - filaments solitary or in
masses, branched or unbranched,
sheathed or unsheathed; specialized
cells may be present
F Important Genera:
Anacystis (Microcystis), Gleotrichia.
Anabaena, Aphanizomenon, Schizothrix, and
Oscillatoria.
IV GREEN ALGAE
This group possesses intracellular structures
(such as chloroplasts) and chlorophyll a and b
in the ratio 2:1, similar to higher plants.
A The cells are grass green because of the
predominance of the chlorophylls over the
carotenes and xanthophylls.
B Sexual reproduction and motile cells are
common.
C These algae are commonly divided into
three groups:
1 Coccoid - non-motile, solitary or
colonial. Actinastrum. Ankistrodesmus,
Coelastrium. Golenkinia. Oocystis,
Pediastrum, Scenedesmus.
2 Filamentous - attached or planktonic,
branched or unbranched, Ulothrix,
Mougeotia. Spirogyra, Stigeoclonium,
Cladophora.
3 Flagellated -2-4 flagella, solitary or
colonial
D Important Genera: Chlamydomonas.
Pandorina, Eudorina. Volvox. Carte ria
V YELLOW-BROWN
These algae are yellowish green to golden
brown because of a predominance of carotenes
and xanthophylls. This group is usually the
dominant form in the plankton and periphyton.
The phylum contains coccoid, filamentous,
and flagellate forms of varying importance.
A Coccoid - includes the diatoms (see below)
B Filamentous - important genera include
Tribonema, Vane her ia
C Flagellates - flagella single or paired,
equal or unequal in length; ceUs solitary
or colonial; some cells are armored.
Important genera: Chromulina,
Chrysococcus. Kephrion. Synura,
Dinobryon.
D Diatoms
This group generally contributes 70-80%
of the phytoplankton and periphyton.
The cell walls contain silica and possess
distinctive markings. The chloroplasts
contain chlorophyll a and £ in nearly
equal amounts, but appear yellow brown
because of the predominance of /3-carotene
and fucoxanthin. The diatoms are
generally divided into two major groups:
1 Centric
The cells are circular in cross-section;
drum-shaped, cylindrical, or spindle-
shaped in side view. Important genera
include: Cyclotella, Stephanodiscus,
Melosira, Rhizosolenia.
2 Pennate
The cells are usually rectangular in
cross-section and boat-shaped, with
transverse rows of linear markings.
Important genera include: Fragilaria,
Synedra. Asterionella. Navicula,
Nitzschia. Gomphonema.
II 3-3
-------
Types of Algae
VI EUGLENOPHYTA
A This group contains chlorophyll ji and b
and consists primarily of motile forms
that have a single flagellum.
B The common genera are Euglena. Phacus.
and Trachelomonas.
X CHLOROMONDOPHYTA
This is a small group of miscellaneous,
little-understood, protozoa-like forms.
There are no common genera in this phylum.
REFERENCES
VII PYRROPHYTA
A These algae are greenish tan to golden
brown, and contain chlorophyll a and £.
The dominant forms (Dinoflagellates) are
unicellular, motile, and are encircled by
a transverse or spiral groove. One of the
two flagella lies in the groove and encircles
the cell; the other extends backward from
the cell. Some forms lack a cell wall,
while others have thick walls which appear
to be composed of many plates.
B The common genera include Gymnodinium,
Glenodinium. Gonyaulax. Peridinium.
and Ceratium.
VIE RHODOPHYTA
A This phylum is largely marine, and contains
chlorophyll a, /3-carotene, phycoerythrin,
and phycocyanin. The common freshwater
species are filamentous, attached forms,
and are not found in the plankton.
B Important freshwater genera include the
filamentous forms, Batrachospernum,
Audouinella and Lemanea.
PHAEOPHYTA
The brown algae are almost entirely marine,
and have no important freshwater forms.
They contain chlorophyll a and c and /3-carotene,
but are brown because of the predominance
of fucoxanthin and other xanthophylls. This
group is usually macroscopic, and contains
the large "sea weeds" or kelp.
1 Davis, C. C. The marine and freshwater
plankton. Mich. State Univ. Press,
East Lansing. 1955.
2 Fritsch, F. E. The structure and
reproduction of the algae. Vol. I.
Cambridge University Press. 1956.
3 Fritsch, F.E. The structure and
reproduction of the algae. Vol. n.
Cambridge University Press. 1965.
4 Palmer, C.M. Algae and water supplies,
USDHEW, PHS, DWSPC, Cincinnati,
1962.
5 Prescott, G.W. How to know the fresh-
water algae. Win. C. Brown Company,
Dubuque. (revision in press) 1954.
6 Prescott, G.W. Algae of the Western
Great Lakes area. 2nd ed. Wm. C.
Brown, Dubuque. 1962.
7 Prescott, G.W. The algae: A Review.
Houghton Mifflin Co., Boston. 1968.
8 Round, F.E. The biology of the algae.
Edward Arnold, Ltd., London. 1965.
9 Smith, G.M. The freshwater algae of
the United States. 2nd ed. McGraw-
Hill Book Co., New York. 1950
10 Tiffany, L.H. and Britton, M.E.
The Algae of Illinois. University
Chicago Press, Chicago (University
Microfilms, Ann Arbor, Xerox). 1952.
This outline was prepared by Dr. C.I. Weber,
Chief, Biological Methods Section, Analytical
Quality Control Laboratory, FWPCA,
1014 Broadway, Cincinnati, OH 45202.
II 3-4
-------
Types of Algae
GREEN ALGAE, NON-FILAMENTOUS
Dictyosphaerium
Polyedriopsi9
II 3-5
-------
Types of Algae
GREEN ALGAE, FILAMENTOUS
Spirogyra
II 3-6
-------
Types of Algae
BLUE-GREEN ALGAE
Coccochlor IB
(Gloeothece)
Anacystis
(Chroococcus)
I.yngbya
Aphanizomenon
Gomphosphae ria
Schizothrix
Calothrix
Agmenellum
(Merismophedia)
n 3-7
-------
Types of Algae
FLAGELLATE ALGAE
Chlorogoniu
Pandorina
II 3-8
-------
Types of Algae
DIATOMS
Navicula
(Girdle View)
Cyclotella
(Valve View)
Cyclotella
(Girdle View)
die View)
Nitzschia
n 3-9
-------
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 (phyocyanm-c) which tends to give the
cushions or mats they may form a blue-green
tinge.
Ill WHERE ARE THE BLUE-GREENS FOUND?
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-
tems. The gelatinous sheath is com-
posed of pectic substances, cellulose
and related compounds.
BI. MIC. cla. ICa. 8. 69
II 4-1
-------
Blue-Green Algae
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.
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-
tenstic 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
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, healthy 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>
No. Healthy blooms are produced by myraids
of cells living near the surface of the water
at times when environmental conditions are
especially 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.
XII 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 not 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.
II 4-2
-------
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 (i. e., not terminal).
4 Akinetes are cylindrical and
relatively long.
II 4-3
-------
Blue-Green Algat;
SOME BLUE-GREEN ALGAE
1. Notvfitamentp.tfs ( cbccoid)..Blue-Green Algae:
:>i-'i;i;l'.''. - ":-. /'/xT
II' >-| . iti'.
-------
Blue-Green Algae
5 Often imparts grassy or nastur-
tium-like odors to water.
D Oscillator la 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
hormongoma only.
5 Live species exhibit "oscillatona"
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
tnchome.
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.
Vegetative cells, heterocysts,
and even the akinetes are broader
than long.
II 4-5
-------
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.
in 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
paramylin (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. sangulnea has red pigment.
6 E. viridis generally favors water
rich in organic matter.
7 IS. 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
II 5-1
-------
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 Chlamydotnonas 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
ceU.
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).
II 5-2
-------
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 odor.
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 (Oinophyceae)
(Figures 14-16). This group is almost
exclusively flagellated and is characterized
by chromatophores which are yellow-brown
in 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".
H 5-3
-------
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. cinctum 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
Baciliariophyceae), 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 Lorica distinct
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 IJ. 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.
II 5-4
-------
Green and Other Figmented Flagellates
(fig 1-13 from Lackey and Callaway)
Euglena
Phacus
Lepocinclis
Trachelomonas
GREEN EUGLENOIDS
^
Chlamydomonas
Pandorina
Chlorogonium
GREEN PHYTOMONADS
io p
Chromulina
11
Synura
Dinobryon
YELLOW CHRYSOMONADS
14
Massartia
15
Peridinium
Ceratium
YELLOW-BROWN DINOFLAGELLATES
E 5-5
-------
Green and Other Pigmented Flagellates
FLAGELLATES
(MASTIGOPHORA)
PLANT FLAGELLATES
(PHYTOMASTIGINA)
CHRYSOMONADINA
CRYPTOMONADINA
PHYTOMONADINA
ANIMAL FLAGELLATES
(Z
OOMASTIGIN
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,
FWPCA, Cincinnati, OH 45226.
5-6
-------
FILAMENTOUS GREEN ALGAE
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.
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).
H 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 ceU( "axial").
2 Clear areas of cell sap ("vacuoles") are
generally present in the cell.
1 Including a few yellow-brown and red algae.
IE REPRODUCTION MAY TAKE PLACE
BY SEVERAL METHODS
A Cell division may occur in all cells or
in certain selected ones.
B Spores called akinetes may be formed.
C Zoo spores (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
II 6-1
-------
Filamentous Green Algae
B Branched forms
Cladophora
Pithopora
Stigeoclonium
Chaetophora
Draparnaldia
Rhizoclonium
Audouinella
Bulbochaete
Nitella
C Specialized and related forms
Schizomeris
Comsopogon
Batrachospermum
Chara
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
H.S 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.
O 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.
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. Monostroma
G Zygnemataceae
Zygnema. Spirogyra. Mougeotia
H De smidiaceae
Desmidium. Hyalotheca
I Tribonemataceae
Tribonema. Bumilleria
J Characeae
Chara. Nitella. Tolypella
E 6-2
-------
Filamentous Green Algae
13. GREENS, FILAMENTOUS
MAMWULDU
It*
II 6-3
-------
Filamentous Green Algae
VIE IDENTIFICATION
A Branching and attenuation are of primary
importance.
B P last ids: 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., Sundaralingam,
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.
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.
II 6-4
-------
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.
n 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 shapeboth 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 Micractinium. 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
II 7-1
-------
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 A nkistrodesmus cells are usually long
and slender, tapering to sharp point at
both ends. They may be straight,
curved, or twisted into loose aggregations.
A nkistrodesmus 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 Hantzschii differ only
in the sharpness of the taper toward
the tips of the cells. The former has
relatively little taper, and the latter,
more.
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.
II 7-2
-------
Coccoid Green Algae
D
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 maybe 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.
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 gellatinized
and greatly expanded mother cell wall.
Cells may be ellipsoidal or almost
cylindrical, cell wall thin, no spines
or other ornamentation. Oocvstis
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. Dimorphococ cus
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
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 in 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.
II 7-3
-------
Coccoid Green Algae
Micrasterias is relatively common,
ornate.
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 oblongum 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.
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 Staurastrum punctulatum is reported
as an indicator of clean water.
c Staurastrum paradoxicum causes a
grassy odor in water.
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.
2 Smith, G.S. Phytoplankton of the
Inland Lakes of Wisconsin. Part I.
Bulletin No. 57, Scientific Series
No. 12. 1920.
in 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.
This outline was prepared by H. W. Jackson,
Chief Biologist, National Training Center,
FWPCA, Cincinnati, OH 45226.
II 7-4
-------
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.
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.
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 cylmdnc ("centric") one view
of which is circular.
1 Pennate diatoms may be symmetrical,
transversely unsymetrical, or longitudi-
nally unsymmetrical.
Ill EXAMPLES OF COMMON DIATOMS:
A Pennate, symmetrical:
Navicula
Pinnulana
Synedra
Nitzschia
Diatoma
Fragilana
Tabellaria
Cocconeis
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.
B Pennate, unsymmetrical:
Gomphonema
Surirella
Cymbella
Achnanthes
Astenonella
Meridiem
C Centric:
D Cell markings include-
1 Raphe or false raphe extending
longitudinally.
2 Striations 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.
Cyclotella
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. lOa. 8.69
II 8-1
-------
Diatoms
Many diatoms are more abundant in late
autumn, winter, and early spring than in
the warmer season.
Fragilaria
Synedra
Asterionella
B The walls of dead diatoms generally remain
undecomposed and may be common in 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.
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
Pinnulana
Stauroneis
Pleurosigma
Amphiprora
Gomphonema
Cymbella
Epithemia
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
Stephanodisc us
Melosira
Rhizosolema
Biddulpnia
B Pennals Group
1 Fragilanneae. The false raphe group.
Representative genera:
Tabe liana
Meridion
Diatoma
4 Sunrellmeae. 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.
II 8-2
-------
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
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 8-3
-------
FILAMENTOUS BACTERIA
I INTRODUCTION
There are a number of types of filamentous
bacteria that occur in the aquatic environment.
They include the sheathed sulfur and iron
bacteria such as Beggiatoa, Crenothrix and
Sphaerotilus, the actinomycetes which are
unicellular microorganisms that form chains
of cells with special branchings, and
Gallionella, a unicellular organism that
secretes a long twisted ribbon-like stalk.
These filamentous forms have at times
created serious problems in rivers, reservoirs,
wells, and water distribution systems.
II BEGGIATOA
Beggiatoa is a sheathed bacterium that grows
as a long filamentous form. The flexible
filaments may be as large as 25 microns
wide and 100 microns long. Transverse
separations within the sheath indicate that a
row of cells is included in one sheath. The
sheath may be clearly visible or so slight that
only special staining will indicate that it is
present.
The organism grows as a white slimy or
felted cover on the surface of various objects
undergoing decomposition or on the surface
of stagnant areas of a stream receiving sewage.
It has also been observed on the base of a
trickling filter and in contact aerators.
It is most commonly found in sulfur springs
or polluted waters where I^S is present.
Beggiatoa is distinguished by its ability to
deposit sulfur within its cells; the sulfur
deposits appear as large refractile globules.
When I^S is no longer present in the environ-
ment, the sulfur deposits disappear.
Dr. Pringsheim of Germany has recently
proved that the organism can grow as a true
autotroph obtaining all its energy from the
oxidation of HgS and using this energy to fix
into all material. It can also use certain
organic materials if they are present along
with the H2S.
Faust and Wolfe, and Scotten and Stokes
have grown the organism in pure culture in
this country. Beggiatoa exhibits a motility
that is quite different from the typical
flagellated motility of most bacteria; the
filaments have a flexible gliding motion.
The only major nuisance effect of Beggiatoa
known has been overgrowth on trickling
filters receiving waste waters rich in I^S.
The normal microflora of the filter was
suppressed and the filter failed to give good
treatment. Removal of the I^S from the
water by blowing air through the water
before it reached the filters caused the
slow decline of the Beggiatoa and a recovery
of the normal microflora. Beggiatoa
usually indicates polluted conditions with
the presence of HgS rather than being a
direct nuisance.
Ill ACTINOMYCETES AND EARTHY ODORS
IN WATER
Actinomycetes are unicellular microorganisms,
1 micron in diameter, filamentous, non-
sheathed, branching monopodially, and re-
produced by fission or by means of special
conidia. Their filamentous habit and method
of sporulation is reminiscent of fungi. How-
ever, their size, chemical composition, and
other characteristics are more similar to
bacteria. These organisms may be con-
sidered as a group intermediate between the
fungi and the bacteria. They require organic
matter for growth but can use a wide variety
of substances and are widely distributed.
Actinomycetes have been implicated as the
cause of earthy odors in some drinking
waters (Romano and Safferman, Silvey and
Roach) and in earthy smelling substance has
BA.8. 1.67
H 9-1
-------
Filamentous Bacteria
been isolated from several members of the
group by Gerber and Lechevalier. Safferman
and Morris have reported on a method for the
"isolation and Enumeration of Actmomycetes
Related to Water Supplies. " But the actmo-
mycetes are primarily soil microorganisms
and often grow in fields or on the banks of a
river or lake used for the water supply.
Although residual clorination will kill the
organisms in the treatment plant or distribu-
tion system, the odors often are present
before the water enters the plant. Use of
permanganate oxidation and activated carbon
filters have been most successful of the
methods tried to remove the odors from the
water. Control procedures to prevent the
odorous material from being washed into the
water supply by rams or to prevent possible
development of the actmomycetes in water
rich in decaying organic matter is still needed.
IV FILAMENTOUS IRON BACTERIA
The filamentous iron bacteria of the
Sphaerotilus-Lepthothrix group, Crenothrix,
and Gallionella have the ability to either
oxidize manganous or ferrous ions to manganic
or ferric salts or are able to accumulate
precipitates of these compounds within the
sheaths of the organisms. Extensive growths
or accumulations of the empty, metallic
encrusted sheaths devoid of cells, have
created much trouble in wells or water dis-
tribution systems. Pumps and back surge
valves have been clogged with masses of
material, taste and odor problems have
occurred, and rust colored masses of
material have spoiled products in contact
with water.
Crenothrix polyspora has only been examined
under the microscope as we have never been
able to grow it in the laboratory. The organ-
ism is easily recognized by its special
morphology. Dr. Wolfe of the University of
Illinois has published photomicrographs of
the organism.
Organisms of the Sphaerotilus-Leptothnx
group have been extensively studied by many
investigators (Dondero e_t_. _al. , Oondero,
Stokes, Waitz and Lackey, Mulder and
van Veen, and Amberg and Cormack.) Under
different environmental conditions the mor-
phological appearance of the organism
varies. The usual form found in polluted
streams or bulked activated sludge is
Sphaerotilus natans. This is a sheathed
bacterium consisting of long, unbranched
filaments, whereby individual rod-shaped
bacterial cells are enclosed in a linear
order within the sheath. The individual cells
are 3-8 microns long and 1. 2-1.8 microns
wide. Sphaerotilus grows in great masses;
at times in streams or rivers that receive
wastes from pulp mills, sugar refineries,
distilleries, slaughterhouses, or milk pro-
cessing plants. In these conditions, it
appears as large masses or tufts attached
to rocks, twigs, or other projections and
the masses may vary in color from light
grey to reddish brown. In some rivers large
masses of Sphaerotilus break loose and clog
water intake pipes or foul fishing nets. When
the cells die, taste and odor problems may
also occur in the water.
Amberg, Cormack, and Rivers and McKeown
have reported on methods to try to limit the
development of Sphaerotilus in rivers by mter-
mittant discharge of wastes. Adequate
control will probably only be achieved once
the wastes are treated before discharge to
such an extent that the growth of Sphaerotilus
is no longer favored in the river. Sphaerotilus
grows well at cool temperatures and slightly
low DO levels in streams receiving these
wastes and domestic sewage. Growth is slow
where the only nitrogen present is inorganic
nitrogen; peptones and proteins are utilized
preferentially.
Gallionella is an iron bacterium which appears
as a kidney-shaped cell with a twisted ribbon-
like stalk emanating from the concavity of the
cell. Gallionella obtains its energy by oxi-
dizing ferrous iron to ferric iron and uses
only CO2 and inorganic salts to form all of
the cell material; it is an autotroph. Large
masses of Gallionella may cause problems
in wells or accumulate in low-flow low-
pressure water mains. Super chlormation
(up to 100 ppm of sodium hypochlorite for 48
hours) followed by flushing will often remove
the masses of growth and periodic treatment
II 9-2
-------
Filamentous Bacteria
will prevent the nuisance effects of the ex-
tensive masses of Galhonella.
REFERENCES
Beggiatoa
1 Faust, L. and Wolfe, R. S. Enrichment
and Cultivation of Beggiatoa Alba.
Jour. Bact. , 81:99-106. 1961.
2 Scotten, H.L. and Stokes, J. L. Isolation
and Properties of Beggiatoa. Arch
Fur. Microbiol. 42:353-368. 1962.
3 Kowalhk, U. and Pringsheim, E.G. The
Oxidation of Hydrogen Sulfide by
Beggiatoa. Amer. Jour, of Botany.
53:801-805. 1966.
Actinomycetes and Earthy Odors
4 Silvey, J.K.G. et_._al. Actinomycetes and
Common Tastes and Odors. JAWWA,
42:1018-1026. 1950.
5 Safferman, R. S. and Morris, M.E. A
Method for the Isolation and Enumera-
tion of Actinomycetes Related to Water
Supplies. Robert A. Taft Sanitary
Engineering Center Techn. Report
W-62-10. 1962.
6 Gerber, N.N. and Lechevalier, H.A.
Geosmin, an Earthy-Smelling Substance
Isolated from Actinomycetes. Appl.
Microbiol. 13:935-938. 1965.
Filamentous Iron Bacteria
7 Wolfe, R. S. Cultivation, Morphology, and
Classification of the Iron Bacteria.
JAWWA, 50:1241-1249. 1958.
8 Kucera, S. and Wolfe, R. S. A Selective
Enrichment Method for Galhonella
ferruginea. Jour. Bacteriol. 74:344
349. 1957.
9 Wolfe, R. S. Observations and Studies
of Crenothnx polyspora. JAWWA,
52:915-918. 1960.
10 Wolfe, R. S. Microbiol. Concentration
of Iron and Manganese in Water with
Low Concentrations of these Elements.
JAWWA, 52:1335-1337. 1960.
11 Stokes, J.L. Studies on the Filamentous
Sheathed Iron Bacterium Sphaerotilus
natans. Jour. Bacteriol. 67:278-291.
1954.
12 Waitz, S. and Lackey, J. B. Morphological
and Biochemical Studies on the Organ-
ism Sp_haerotilu£ natans. Quart. Jour.
Fla. Acad. Sci., 21(4):335-340. 1958.
13 Dondero, N.C., Philips, R.A., and
Henkelkian, H. Isolation and Preser-
vation of Cultures of Sphaerotilus.
Appl. Microbiol. 9:219-227. 1961.
14 Dondero, N.C. Sphaerotilus, Its Nature
and Economic Significance. Advances
Appl. Microbiol. 3:77-107. 1961.
15 Mulder, E.G. and van Veen, W.L. Inves-
tigations on the Sphaerotilus -Leptothrix
Group. Antonie van Leewenhoek, 29:
121-153. 1963.
16 Amberg, H. R. and Cormack, J.F. Factors
Affecting Slime Growth in the Lower
Columbia River and Evaluation of Some
Possible Control Measures. Pulp and
Paper Mag. of Canada, 61:T70-T80.
1960.
17 Amberg, H.R., Cormack, J.F. and
Rivers, M. R. Slime Growth Control
by Intermittant Discharge of Spent
Sulfite Liquor. Tappi, 45:770-779.
1962.
18 McKeown, J.J. The Control of Sphaerotilus
natans. Ind. Water and Wastes, 8:(3)
19-22 and 8:(4)30-33. 1963.
This outline was prepared by R. F. Lewis,
Bacteriologist, Microbiological Activities,
Research and Development, Cincinnati Water
Research Laboratory, FWPCA.
II 9-3
-------
FUNGI
I INTRODUCTION
A Description
Fungi are heterotrophicachylorophyllous
plant-like organisms which possess true
nuclei with nuclear membranes and nu-
cleolL. 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 i
Phylum Mycota of the plant kingdom.
Some authorities consider the fungi an
essentially monophyletic group distinct
from the classical plant and animal
kingdoms.
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.
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 microaerophihc metabolism,
but anaerobic catabolism is not uncommon.
A few species show anaerobic metabolism
and growth.
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.6.4.69
II 9-5
-------
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- l?n)
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 tri-
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 truefungalhyphae.
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).
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
(i.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 Emerson and
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
II 9-6
-------
PLATE I
"SEWAGE FUNGUS" COMMUNITY
(Attached "filamentous" growths)
Fungi
Zoogloea
Sphaerotilus natans
Beggiatoa alba
BACTERIA
Fusarium aqueductum
Leptomitus lacteus
Geotrichum candidum
FUNGI
// Epistylis 8
10
Opercularia
PROTOZOA
-------
Fungi
PLATE II
REPRESENTATIVE FUNGI
Figure *
Ftuarium aquacductuum
(Radlmacher and
Rabcnhorst) Saccardo
Microconidia (A) produced
from phialides u in Ctphalo-
iporium, remaining in ali:
balls. Macroconidia (B), with
one to several cross walls,
produced from collared phial-
ides. Drawn from culture.
Figure 3
Gcotrichiun candidum
Link ex Persoon
Mycelium -with short cells
and arthrospores. Young hy-
pha (A); and mature arthro-
spores (B). Drawn from cul-
ture.
Figure O
Achlya americana Humphrey
Ooogonium with three oo-
spores (A); young zoospor-
angium with delimited zoo-
spores (B) ; and zoosporangia
(C) with released zoosporcs
that remain encysted in clus-
ters at the month of the dis-
charge tube. Drawn from cul-
ture.
Leptomitiu lacleia (Roth)
Agardh
Cells of the hyphae show-
ing constrictions with cellulin
plugs. In one cell large zoo-
spores have been delimited.
Redrawn from Coker, 1923.
Mycelium with hypbal pegs
(A) on which rotifers will
become impaled; gemmae (B)
produced as conidia on short
hyphal branches; and rotifer
impaled on hyphal peg (C)
from which hyphae have
grown into the rotifer whose
shell will be discarded after
the contents are consumed.
Drawn from culture.
B.
FIGURE / Haplosporidivm costale. Amature spore;
Bearly plnsmodimn.
Figures 1 through 5 from Cooke; Figures 6 and 7 from Galtsoff.
-------
Fungi
sugars and grows most luxuriantly
in the presence of organic nitro-
genous wastes.
3 Ecological Roles
Although the "sewage fungi" on
occasion attain visually notice-
able concentrations, the less
obvious populations of deutero-
mycetes may be more important
in the ecology of the aquatic
habitat. Investigations of the
past decade indicate that nu-
merous 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.
basis of the morphology of the sexual and
zoosporic stage s. In practical schematics,
however, numerous fungi do not demon-
strate these stages. Classification must
therefore be based on the sum total of the
morphological and/or physiological char-
acteristics. 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 pre-
sented on the following pages.
IV Classification
In recent classification schemes, classes of
fungi are distinguished primarily on the
This outline was prepared by Dr. Donald G.
Ahearn, Professor of Biology, Georgia
State College, Atlanta. Georgia. 30303
II 9-9
-------
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
I1 Cell walls usually well defined, somatic phase not 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 comdia-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
mycehal and very extensive. The outstanding characteristics of the thallus is a tendency
to be nonseptate and, in most groups, multinuchate, cross walls are laid down in vigorously
growing material 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 (I1) Hyphal filaments when present septate, without zoospores. with or without sporangia,
usually with conida, 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 .Chytridiomycetes
The Chytridiomycetes 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 (lack 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
Dermocvstidium 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
. Hyphochytridiomycetes
'Lliesc fungi are aquatic (fresh water or marine) chytrid-hke fungi whose motile cells
possess a single anterior flagellum of the tinsel type (feather-like) They are parasitic on
algae and fungi or may be saprobic. Cell walls contain chitin with some species also demon-
stiating 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 cocnocytic, 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 parasitical Members of the family Saprolegniaceac are the common
-------
Fungi
'water molds' and are among the most ubiquitous fungi in nature The order Lagemdiales
includes only a few species which are parasitic on algae small animals, and other aquatic
life The somatic structures of this taxon are holocarpic and cndobiotic The sewage fungi
are classified in the order Leptomitales 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 lacleus may
produce rather extensive fouling floes 01 slimes in organically enriched waters
6' Flagella of unequal size, both uhiplash class Plasmodiophoromycetcs
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 these fungi apparently lack cellulose
7 (3') Mainly'saprobic, sex cell when present a zygospore class 7.\ tiomycetes
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 aie
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-hke 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)
71 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 iji 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 (21) 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 Hemiascomycetidae 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
Barren 1968)
-------
Fungi
KEY TO THE FORM-ORDERS OF THE FUNGI IMPERFECT l
1 Reproduction by means of conidia, oidia, or by budding 2
I1 No reproductive structures present . . . Mycelia Sterilia
2 (1) Reproduction by means of conidia borne in pycnidia Sphaeropsidales
2' Conidia, when formed, not in cycnidia 3
3 (21) Conidia borne in acervuh Melanconiales
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, arthrospores occasionally produced, manifest melanin pigmentation lacking 2
1' Thallus mainly filamentous, dark melanin pigments sometimes produced 3
2 (1) Ballistospores produced Sporobolomycetaceae
21 No ballistospores Cryptococcaceae
3 Comdiophores, if present, not united into sporodochia or synnemata 4
3' Sporodochia present . Tuberculanaceae
3" Synnemata present Stilbellaceae
4 (3) Conidia and conidiophores or oidia hyaline or brightly colored Monihaceae
4' Conidia and/or conidiophores, containing dark melanin pigment Dematiaceae
-------
Fungi
SELECTED1 REFERENCES
Ahearn, D.G., Roth, F.J. Jr., Meyers, S.P.
Ecology and Charact erization of Yeasts
from Aquatic Regions of South Florida.
Marine Biology 1:291-308. 1968
Alexopoulos, J. C. Introductory Mycology.
2nd ed. John Wiley and.Soms, 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
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. InAinsworth, G. C. and
Sussman, A.S. The Fungi, III.
Academic Press, New York. 1968
and Sparrow, F.K., Jr. Fungi
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.
UtahAcad. 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
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.
1187pp. 1960.
. Ecology of Freshwater Fungi.
pp. 41-93. InAinsworth, 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. Bactenol. 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
II 9-13
-------
PROTOZOA, NEMATODES, AND ROTIFERS
I GENERAL CONSIDERATIONS
A Microbial quality constitutes only one
aspect of water sanitation; microchemicals
and radionuclides are attracting increasing
amount of attention lately.
B Microbes considered here include bacteria,
protozoa, and microscopic metazoa; algae
and fungi excluded.
C Of the free-living forms, some are
members of the flora and fauna of surface
waters; others washed into the water from
air and soil; still others of wastewater
origin.
D Hard to separate "native" from "foreign"
free-living microbes, due to close
association of water with soil and other
environments; generally speaking.bacteria
adapted to water are those that can grow
on very low concentrations of nutriment
and zoomicrobes adapted to water are
those that feed on algae.
E More species and lower densities of
microbes in clean water and fewer species
and higher densities in polluted water.
F Pollution-tolerance or nontole ranee of
microbes closely related to the DO level
required in respiration.
G From pollution viewpoint, the following
groups of microbes are of importance:
Bacteria, Protozoa, Nematoda, and
Rotifer a.
II BACTERIA
A No ideal method for studying distribution
and ecology of bacteria in freshwater.
B According to Collins, ^ Pseudomonas,
Achrombacter. Alcaligenes. Chromobac-
terium. Flavobacterium. and Micrococcus
are the most widely distributed and may be
considered as indigenous to natural
waters. Sulfur and iron bacteria are
more common in the bottom mud.
C Actinomycetes, Bacillus spp., Aerogenes
spp., and nitrogen-fixation bacteria are
primarily soil dwellers and may be washed
into the water by runoffs.
D E. coli, streptococci, and_Cl. perfringens
are true indicators of fecal pollution.
HI PROTOZOA
A Classification
1 Single-cell animals in the most
primitive phylum (Protozoa) in the
animal kingdom.
2 A separate kingdom. Protista, to in-
clude protozoa, algae, fungi, and
bacteria proposed in the 2nd edition
of Ward-Whipple's Fresh-Water
Biology.(10)
3 Four subphyla or classes:
a Mastigophora (flagellates) - Sub-
class Phytomastigina dealt with '
under algae; only subclass Zoo-
mastigina included here; 4 orders:
1) Rhizomastigina - with flagellum
or flagella and pseudopodia
2) Protomonadina - with 1 to 2
flagella; mostly free-living many
parasitic
3) Polymastigma - with 3 to 8 flagella,
mostly parasitic in elementary tract
of animals and man
4) Hypermastigina - all inhabitants
of elementary tract of insects
W.BA.45b. 11.67
n 10-1
-------
Protozoa, Nematodes, and Rotifers
Ciliophora or Infusoria (ciliates) -
no pigmented members; 2 classes:
1) Ciliata - cilia present during the
whole trophic life; containing
majority of the ciliates
2) Suctoria - cilia present while
young and tentacles during trophic
life.
Sarcodina (amoebae) - Pseudopodia
(false feet) for locomotion and food-
capturing; 2 subclasses:
1) Rhizopoda - Pseudopodia without
axial filaments; 5 orders:
a) Proteomyxa - with radiating
pseudopodia; without test or
shell
b) Mycetozoa - forming plasmodium;
resembling fungi in sporagium
formation
c) Amoebina - true amoeba -
forming lobopodia
d) Testacea - amoeba with single
test or shell of chitinous
material
e) Foraminifera - amoeba with 1
or more shells of calcareous
nature; practically all marine
forms
Sporozoa - no organ of locomotion;
amoeboid in asexual phase; all
parasitic
B General Morphology
1 Zoomastigina:
Relatively small size (5 to 40 u); with
the exception of Rhizomastigina, the
body has a definite shape (oval, leaf-
like, pear-like, etc.); common members
with 1 or 2 flagella and some with 3, 4,
or more; few forming colonies; c/i ;stome
present in many for feeding.
2 Ciliophora:
Most highly developed protozoa; with
few exceptions, a macro and a micro-
nucleus; adoral zone of membranellae,
mouth, and groove usually present in
swimming and crawling forms, some
with conspicuous ciliation of a disc-like
anterior region and little or no body
cilia (stalked and shelled forms);
Suctoria nonmotile (attached) and with-
out cytostome cysts formed in most.
3 Sarcodina:
Cytoplasmic membrane but no cell wall;
endoplasm and ectoplasm distinct or in-
distinct; nucleus with small or large
nucleolus; some with test or shell;
moving by protruding pseudopodia; few
capable of flagella transformation; fresh-
water actinopods usually sperical with
many radiating axopodia; some Testacea
containing symbiotic algae and mistaken
for pigmented amoebae; cysts with single
or double wall and 1 or 2 nuclei.
4 Sporozoa: to be mentioned later.
C General Physiology
1 Zoomastigina-
Free-living forms normally holozic;
food supply mostly bacteria in growth
film on surfaces or clumps relatively
aerobic, therefore the first protozoa to
disappear in anaerobic conditions and
re-appearing at recovery; reproduction
by simple fission or occasionally by
budding.
2 Ciliophora:
Holozoic; true ciliates concentrating
food particles by ciliary movement
around the mouth part; suctona sucking
through tenacles; bacteriaand small
II 10-2
-------
Protozoa, Nemaiodes, and Rotifers
algae and protozoa constitute main
food under natural conditions; some
shown in laboratory to thrive on dead
organic matter and serum protein; not as
aerobic as flagellates - some surviving
under highly anaerobic conditions, such
as Metopus; reproduction by simple
fission, conjugation or encystation.
3 Sarcodina:
Holozoic; feeding through engulfing by
pseudopodia; food essentially same as
for ciliates; DO requirement somewhat
similar to ciliates - the small amoebae
and Testacea frequently present in large
numbers in sewage effluent and polluted
water; reproduction by simple fission
and encystation.
IV NEMATODES
A Classification
1 All in the phylum Nemata (nonsegment-
ed round worms); subdivided by some
authors into two classes:
Secernentea - 3 orders:
(phasmids)
Tylenchida, Rhabditida, Strongylida,
and Teratocephalida, with papillae on
male tail, caudal glands absent.
Adenophora - 6 orders:
(aphasmids)
Araeolaimida, Dorylaimida,
Chromdonda, Monhysterida, Enoplida,
and Trichosyrmgida no papillae on
male trail, caudal glands absent.
Orders encountered in water and sewage
treatment - Free-living forms inhabit at-
ing sewage treatment plants are usually
bacteria-feeders and those feeding on
other nematodes; those inhabitating clean
waters feeding on plant matters; they
fall into the following orders:
Tylenchida - Stylet in mouth; mostly
plant parasites; some feed on
nematodes, such as Aphelenchoides.
Rhabditida - No stylet in mouth, caudal
glands present, mostly bacteria-feeders,
common genera: Rhabiditis, Diplogaster,
Diplogasteroides, Monochoides, Pelodera,
Panagrellus, and Turbatrix.
Dorylaimida - Relatively large nematodes;
stylet in mouth, feeding on other nematodes
and probably zoomicrobes; Dorylaimus
common genus.
Chromadorida - Many marine forms,
some freshwater dwellers feeding on
algae; characterized by strong orna-
mentation of knobs, bristles or
punctations in cuticle.
Monhysterida - Freshwater dwellers;
esophago-intestinal valve spherical to
elongated, ovaries single or paired,
usually straight; common genus in
water - Monhystera.
Enoplida - Head usually with a number
of setae, Cobb reported one genus,
Mononchulus, in sand filters in
Washington, D.C.
B General Morphology
Round, slender, nonsegmented(transverse
markings in cuticle of some) worms;
some small (about | mm long, as Tn-
cephalobus), many 1 to 2 mm long
(Rhabditis. Diplogaster. and Diplogasteriodes
for instance), and some large (2 to 7 mm,
such as Dorylaimus); sex separated but few
parthenogenetic; complete elementary canal
with elaborate mouth parts with or without
stylet, complete reproductive system in
each sex, no circulatory or respiratory
system; complex nervous system with
conspicuous nerve ring across oesophagus.
C General Physiology
1 Feeding - Most sewage treatment plant
dwellers feeding on bacteria; others
preying on protozoa, nematodes, rotifers.
II 10-3
-------
Protozoa, Nematodes, and Rotifers
etc., clean-water species apparently
vegetarians; those with stylet in mouth
use the latter to pierce the body of animal
or plant and suck contents; metabolic
waste mostly liquid containing ammonium
carbonate or bicarbonate; enteric
pathogens swallowed randomly with
suspending fluid, hence remote possi-
bility of sewage effluent-borne nematodes
being pathogen-carriers.
2 Oxygen requirement - DO apparently
diffused through cuticle into body; DO
requirement somewhat similar to
protozoa, Rhabditis tolerating reduced
DO better than other Rhabditida members;
all disappear under sepsis in liquid; some
thrive in drying sludge.
3 Reproduction - Normal life cycle requires
mating, egg with embryo formation,
hatching of eggs inside or outside femals,
4 larval stages, and adult; few repro-
duce in the absence of males.
V ROTIFERS
A Classification:
1 Classified either as a class of the phylum
Aschelminthes (various forms of worms)
or as a separate phylum (Rotifera); com-
monly called wheel animalcules, on
account of circular 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 (Seisomdeaand
Bdelloidea) grouped by some authors
under Digononta (2 ovaries) and the
other being Monogononta (1 ovary);
Seisonidea containing mostly marine
forms.
3 Class Digononta containing 1 order
(Bdelloida) with 4 families, Philodmedae
being the most important.
4 Class Monogononta comprising 3 orders:
Notommatida (mouth not near center of
corona) with 14 families, Floscularida
Melicertida (corona with two wreaths of
cilia and furrow between them) with 3
families; most import genera included
in the order Notommatida: Brachionus,
H 10-4
Keratella, Monostyla. Trichocerca,
Asplanchna, Polyarthra. Synchaeta.
Microcodon; common genera under the
order Flosculariaceae: Flosculana,
and Atrochus. Common genera under
order Melicertida: Limnias and
Conochilus.
5 Unfortunately orders and families of
rotifers partly 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, connected 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 Philodma, concentrating
bacteria and other microbes and minute
particulate organic matter by ciliary
movement on corona larger microbes
chewed by mastax; some such as
Monostyla 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.
-------
Protozoa. Nenv.todes, and Rotifers
VI SANITARY SIGNIFICANCE
A Pollution tolerant and pollution non-
tolerant species - hard to differentiate -
requiring specialist training in protozoa,
nematodes. and rotifers.
B Significant quantitative difference in clean
and polluted waters - clean waters con-
taining large variety of genera and species
but quite low in densities.
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 enertic pathogens.
VII EXAMINATION OF WATER FOR MICROBES
A Bacteria - not dealt here.
B Protozoa and rotifers - should be included
in examination for planktonic microbes.
C Nematodes
(3)
D Laboratory Apparatus
1 Sample Bottles - One-gallon glass or
plastic bottles with metal or plastic
screw caps, thoroughly washed and
rinsed three times with distilled water.
2 Capillary Pipettes and Rubber Bulbs -
Long (9 in.) Pasteur capillary pipettes
and rubber bulbs of 2 ml capacity.
3 Filtration Unit - Any filter holder
assembly used in bacteriological
examination.'^' The funnel should be
at least 650 ml and the filter flask at
least 2 liter capacity.
4 Filter Membranes - Millipore SS (SS
047 MM) type membranes or equivalent.
5 Microscope - Binocular microscope
with 10X eyepiece, 4X, 10X, and 43X
objectives, and mechanical stage.
E Collection of Water Samples
Samples are collected in the same manner
as those for bacteriological examination,' '
except that a dechlormating agent is not
needed. One-half to one gallon samples
are collected from raw water and one-gallon
samples from tap water. Refrigeration is
not essential and samples may be trans-
ported without it unless examination is to
be delayed for more than five days.
F Concentration of Samples
1 One gallon of tap water can usually be
filtered through a single membrane
within 15 minutes unless the water has
turbidity. At least one gallon of sample
should be used in a single examination.
Immediately after the last of the water
has disappeared through the membrane,
the suction line is disconnected and the
membrane placed on the wall of a clean
50 to 100 ml beaker and flushed re-
peatedly with about 2 ml of sterile
II 10-5
-------
Protozoa, Nematodes, and Rotifers
distilled water with the aid of a capillary
pipette and a rubber bulb. The concen-
trate is then pipetted into a clean
Sedgewick-Rafter Counting Cell and is
ready for examination.
2 In concentration of raw water samples
having visible turbidity, two to four
membranes may be required per sample,
with filtration through each membrane
being limited to not more than 20
minutes. Samples ranging from 500 ml
to 2 liters may be filtered, depending on
whether turbidity is high or low. After
filtration the membranes are placed on
the walls of separate beakers and
washed as above. To prevent the parti-
culates from obscuring the nematodes,
the washing from each filter is examined
in a separate counting chamber.
G Direct Microscopic Examination
Each counting chamber containing the
filter concentrate is first examined under
a 4X objective. Unless the concentrate
contains more than 100 worms, the whole
cell area is surveyed for nematodes, with
respect to number, developmental stage,
and motility. When an object having an
outline resembling that of a nematode is
observed, it is re-examined under a 10X
objective for anatomical structures, unless
the object exhibits typical nematode move-
ment, which is sufficient for identifying the
object as a nematode. When the concentrate
contains more than 100 worms, the worm
density can be estimated by counting the
number of worms in representative micro-
scopic fields and multiplying the average
number of worms per field by the number
of fields in the cell area. The nematode
density may be expressed as number of
worms per gallon with or without differenti-
ation as to adult or larval stages or as to
viability.
H General Identification of Nematodes
1 While actively motile nematodes can be
readily recognized by any person who
has some general concept of micro-
scopic animals, the nonmotile or
VIII
sluggishly motile nematodes may be
confused with root fibers, plant fila-
ments of various types, elongated
ciliates such as Homalozoon vermi-
culare. or segments of appendages of
small Crustacea. To facilitate a
general identification of nematodes, the
gross morphology of three of the free-
living nematodes that are frequently
found in water supplies is shown in the
attached drawing. The drawing provides
not only the general anatomy for recogni-
tion of nematodes but also most of the
essential structures for guidance to those
who want to use the "Key to Genera" in
chapter No. 15 on Nemata by B. G.
Chitwood and M. W. Allen in the book,
Fresh Water Biology. (10>
2 Under normal conditions, practically
all nematodes seen in samples of
finished water are in various larval
stages and will range from 100 to 500
microns in length and 10 to 40 microns
in width. Except in the fourth (last)
stage, the larvae have no sexual organs
but show other structural characteristics.
3 If identification of genera is desired,
the filter washings are centrifuged at
500 rpm for a few minutes. The
supernate is discarded, except a few
drops, and the sediment is resuspended
in the remaining water. A drop of the
final suspension is examined under both
10X and 43X objectives for anatomical
characteristics without staining, and for
supplementary study of structures the
rest is fixed in 5% formalin or other
fixation fluid and stained according to
instructions given in Chitwood and
Allen's Chapter on Nemata, W
Goodey's Soil and Freshwater Nema-
or other books on nematology.
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.
H 10-6
-------
Protozoa, Nematodes, and Rotifers
Can use them on a quantitative basis -
nematodes, rotifers, and nonpigmented
protozoa present in small numbers in
clean water. Numbers greatly increased
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.
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 pol-
lution 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.
K CONTROL
A Chlorination of effluent
B Prolongation of detention time of effluent
C Elimination of slow sand filters in
nematode control.
LIST OF COMMON ZOOLOGICAL ORGANISMS
FOUND IN SEWAGE TREATMENT PROCESS -
TRICKLING FILTERS
PROTOZOA
Sarcodina - Amoebae
Amoeba proteus, A radios a
Hartmannella Spp
Arcella Vulgaris
Noegleria gruberi
Actmophrys Sol
FLAGELLATA
Bodo caudatus
Pleuromonas jaculans
Oikomonas termo
Cercomonas longicauda
Peranema trichophorium
Swimming type
Ciliophora:
Colpidium colpoda
Colpoda cuculus
Glaucoma pyriforinis
Paramecium candatum; 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)
Crawling type
Euplotes patella
Stylonychia mylitus
Urostyla Spp.
Oxytricha Spp.
NEMATODA
Diplogaster Spp. Rhabditolaimus Sp.
Monochoides Spp. Monhystera Sp.
Diplogasteroides Spp. Trilobus Sp.
Rhabditis Spp.
Pelodera Spp.
Aphelenchoides Sp.
Dorylamus Sp.
Cylmdrocorpus Sp.
Cephalobus Sp.
II 10-7
-------
Protozoa. Nematodes, and Rotifers
ROTATORIA
Diglena
Monstyla
Polyarthra
Philodina
Keratella
Brachionus
OLIGOCHAETA (bristle worms)
Aelosoma hemprichi
Aulophorus limosa
Tubifex tubifex
Lumbricillus lineatus
INSECT LARVAE
Chironomus
Psychoda Spp. (trickling filter fly)
ARTHROPODA
Lessertia Sp.
Porrhomma Sp.
Achoratus subuiaticus (collembola)
Folsomia Sp. (collembola)
Tomocerus Sp. (collembola)
3 Chang, S. L Proposed Method for Examina-
tion of Water for Free-Living Nematodes.
J. A.W. W.A. 52:695-698. i960.
4 Chang, S. L , et al. Survival and Protection
Against Chlorination of Human Enteric
Pathogens in Free-Living Nematodes
Isolated from Water Supplies Am Jour
Trop. Med. and Hyg. 9:136-142 1960
5 Chang, S. L. Growth of Small Free-Living
Amoebae in Bacterial and Bacteria-Free
Cultures. Can. J. Microbial.
6:397-405. 1960
6 Chang, S. L. and Kabler, P.W Free-Living
Nematodes in Aerobic Treatment Plant
Effluents. J. W. P. C.F. 34:1256-1261
1963.
7 Chitwood, B G. and Chitwood, M.B. An
Introduction to Nematology SectionT:
Anatomy 1st ed. Monumental Printing
Co. Baltimore. 1950. pp 8-9.
8 Cobb, N. A. Contributions to the Science
of Nematology VII. Williams and
Wilkins Co. Baltimore. 1918
9 Collins, V G. The Distribution and Ecology
of Bacteria in Freshwater, Pts I & II
Proc. Soc. for Water Treatment and
Exam. 12:40-73. 1963. (England)
10 Edmondson, W. T , et al. Ward-Whipple's
Fresh Water Biology. 2nd ed. John
fork
Wiley It Sons, New
pp 368-401.
1959
11
Goodey, T. Soil and Freshwater Nematodes
(A Monograph) 1st ed. ivietnuen and Co.
Ltd. London 1951.
REFERENCES
1 American Public Health Association,
American Water Works Association and
Water Pollution Control Federation.
Standard Methods for the E xamination
of Water ana wastewater7 J..un ea.
New York.
This outline was prepared by S. L Chang, Chief,
Etiology, Disease Studies & Quality Control
Laboratories, Water Supply & Sea Resources
Program, National Center for Urban & Industrial
Health.
2 Chang, S. L. , et al. Survey of Free-
Living Nematodes and Amoebas in
Municipal Supplies. J. A. W. W. A. 52:
613-618.
II 10-8
-------
Protozoa, Nematodes, and Rotifers
Effluent
Insects
Oligochaetes &
insect larvae
Nematodes
& rotifers
4-4
Nonpigmented
protozoa *
I I I fit
Heterotrophic
bacteria
Fungi
Algae
Autotrophic bacteria
Pathogenic organisms"
Suspended organic matter
(by hydrolysis)
Dissolved organic matter
(respiration,
deamination,
decarboxylation, etc.)
Raw Sewage
Inorganic C, P, N,
S comp.
(NH3> NO". C0°, P)
(Nitrification, sulfur
& iron bacteria)
Food Chain in Aerobic Sewage Treatment Processes
II 10-9
-------
FREE-LIVING AMOEBAE AND NEMATODES
I FREE-LIVING AMOEBAE
A Importance of Recognizing Small,
Free-Living Amoebae in Water
Supplies
1 Commonly found in soil and
natural, fresh waters - hence,
frequently encountered in ex-
amination of raw water.
2 Cysts not infrequently found in
municipal supplies - not patho-
gen carriers.
3 Cysts not to be confused with
those of Endamoeba histolytica
in water-borne epidemics.
4 Cysts useful in evaluation water
treatment efficiency in remov-
ing or destroying cysts of path-
ogenic amoeba.
B Classification of Small, Free-Living
Amoebae
1 Recognized classification based
on characteristics in mitosis.
2 Common species fall into the
following families and genera:
Family Schizopyrenidae: Gen-
era Naegleria, Didascalus. and
Schizopyrenus - first two being
flagellate amoebae.
Family Hartmannellidae:
Genera Hartmenella (Acantha-
moeba)
3 How to prepare materials for
studying mitosis - Feulgen
stain
C Morphological Characteristics of
Small, Free-Living Amoebae
1 Morphology of trophozoites -
Ectoplasm and endoplasm with
nucleus
2 Morphology of cysts - Single
BI.AQ. 14a.4.70
or double wall with or without
pores
D Cultural Characteristics of Small,
Free-Living Amoebae
1 How to cultivate these amoebae
2 Growth characteristics on plate
cultures
3 Complex growth requirements
for most of these amoebae
E Resistance of Amoebic Cysts to
Physical and Chemical Agents
F Practical Use of Small, Free-Living
Amoebae as Testing Organisms H.
rhysodes. however, can grow in or-
dinary broth.
1 Culture-induced cysts for study-
ing removal efficiency of floc-
culation and sand filtration and
cysticidal efficiency of water
disinfectants.
II FREE-LIVING NEMATODES
A Classification of Those Commonly
Found in Water Supplies
1 Phasmidia: Genera Rhabditis,
Diplogaster. Diplogasteroides.
Cheilobus, Panagrolaimus
2 Aphasmidia: Genera Monhystera,
Aphelenchus, Turbatrix (vinegar
eel). Dorylaimus, and Rhabdol-
aimus
B Morphological Features
1 Phassids
2 Aphasmids
C Life Cycle
1 Methods of mating
2 Stages of development
II 11-1
-------
Amoebae and Nematodes in Water Supplies
3 Parthenogenesis
D Cultivation
1 Bacteria-fed cultures
2 Axenic cultures
E Occurrence in Water Supplies
1 Relationship between their
appearance in finished water
and that in raw water.
2 Frequency of occurrence in
different types of raw water
and sources .
3 Survival of human enteric path-
ogenic bacteria and viruses in
nematodes.
4 Protection of human enteric
pathogenic bacteria and viruses
in nematode-carriers.
G Control
1 Chlorination of sewage effluent
2 Floeculation and sedimentation
of water
3 Chlorination of water
4 Other methods of destruction
REFERENCES
Amoebae
1 Singh, B. N., "Nuclear Division in Nine
Species of Small, Free-Living Amoe-
bae and its Bearing on the Classifica-
tion of the Order Amoebida", Philos.
Trans. Royal Soc. London, Series B,
236:405-461, 1952.
2 Chang, S. L., et al. "Survey of Free-
Living Nematodes and Amoebas in
Municipal Supplies". J.A.W.W.A.
52^:613-618, 1960.
3 Chang, S. L., "Growth of Small Free -
Living Amoebae in Various Bacterial
and in Bacteria-Free Cultures". Can.
Jour. Microbiol. 6:397-405, 1960.
Nematodes
1 Goodey, T., "Freshwater Nematodes",
1st. Edition, Methuen & Co., London,
1951.
2 Edmondson, W. T.. Ed., Ward & Whipple's
"Fresh-Water Biology" 10th Edition,
page 397, 1955.
3 Chang. S. L., etal., "Occurrence of a
Nematode Worm in a City Water Supply".
J.A.W.W.A., £1:671-676, 1959.
4 Chang, S. L., etal., "Survival, and
Protection Against Chlorination, of
Human Enteric Pathogens in Free-
Living Nematodes Isolated From Water
Supplies". Am. Jour. Trop. Medicine
& Hygiene, £:136-142, 1960.
5 Chang, S. L., etal., "Survey of Free -
Living Nematodes and Amoebas in
Municipal Supplies". J.A.W.W.A.,
52:613-618, 1960.
6 Chang, S. L., "Proposed Method for
Examination of Water for Free-Living
Nematodes". J.A.W.W.A., 52:695-698,
1960.
7 Chang, S. L., "Viruses, Amoebas,
and Nematodes and Public Water
Supplies". J.A.W.W.A., 53:288-296,
1961.
8 Chang, S. L., and Kabler, P. W., "Free-
Living Nematodes in Sewage Effluent
from Aerobic Treatment Plants". To
be published.
This outline was prepared by Shih L. Chang,
M.D., Chief, Etiology, Disease Studies &
Quality Control Laboratories, Water Supply
& Sea Resources Program, NCUIH, SEC.
II 11-2
-------
ANIMAL PLANKTON
I INTRODUCTION
A Planktonic animals or zooplankton are
found in nearly every major group of
animals.
1 Truly planktonic species (euplankton)
spend all or most of their active life
cycle suspended in the water. Three
groups are predominantly involved in
fresh water; the protozoa, rotifers,
and microcrustacea.
2 Transient planktonic phases such as
floating eggs and cysts, and larval
stages occur in many other groups.
B Many forms are strictly seasonal in
occurrence.
C Certain rare forms occur in great numbers
at unpredictable intervals.
D Techniques of collection, preservation,
and identification strongly influence the
species reported.
E In oceanographic work, the zooplankton is
considered to include many relatively large
animals such as siphonophores, ctenophores,
heptcropods, pteropods, arrowworms, and
euphausid shrimp.
F The plant-like or phytoplankton on the
other hand are essentially similar in all
waters, and are the nutritional foundation
for the animal community.
II PHYLUM PROTOZOA
A The three typically free living classes,
Mastigophora, Rhizopoda, and Ciliophora,
all have planktonic representatives. As
a group however, the majority of the phylum
is benthic or bottom-loving. Nearly any
of the benthic forms may occasionally be
washed up into the overlying waters and
thus be collected along with the euplankton.
B Class mastigophora, the nonpigmented
zooflagellates.
These have frequently been confused with
the phytomastigina or plant-like flagellates.
The distinction is made here on the basis
of the presence or absence of chlorophyll
as suggested by Palmer and Ingram 1955.
BI.AQ.20c. 4. 70
(Note Figure: Nonpigmented, Non-Oxygen
Producing Protozoan Flagellates in the
outline Oxygen Relationships.)
1 Commonly encountered genera
Bodo
Peranema
2 Frequently associated with eutrophic
conditions
C Class Rhizopoda - amoeboid protozoans
1 Forms commonly encountered as
plankton:
Chaos
Arcella
Difflugia
Euglypha
(Amoeba)
Centropyxis
Heliozoa
2 Cysts of some types may be encountered
in water plants or distribution systems;
rarely in plankton of open lakes or
reservoirs.
D Class Ciliophora
1 Certain "attached" forms often found
floating freely with plankton:
Vorticella
Carchesium
2 Naked, unattached ciliates. Halteria
one of commonest in this group! Various
heavily ciliated forms (holotrichs) may
occur from time to time such as
Colpidium, Enchelys, etc.
3 Ciliates protected by a shell or test
(testaceous) are most often recorded
from preserved samples. Particularly
common in the experience of the National
Water Quality Sampling Network are:
Codonella fluviatile
Codonella cratera
Tintinnidium (usually with organic matter)
Tintinnopsis
II 12-1
-------
Animal Plankton
III PHYLUM ROTIFERA
A Some forma such as Anuraea cochlearis
and Asplanchna pridonta tend to be present
at all dmes of the year. Others such as
Notholca striata, N. longispina and Poly-
artnra piatyptera are reported to be essen-
tiany winter torms.
B Species in approximate order of descending
frequency currently recorded by National
Water Quality Sampling Network are:
Keratella cochlearis
Polyarthra vulgaris
Synchaeta pectinata
Brachionus quadridentata
Trichocerca longiseta
Rotaria sp.
Filinia longiseta
Kellicottia longispina
Pompholyx sp.
C Benthic species almost without number may
be collected with the plankton from time to
time.
IV PHYLUM ARTHROPODA
A Class Crustacea
1 The Class Crustacea includes the larger
common freshwater euplankton. They
are also the greatest planktonic consum-
ers of basic nutrients in the form of
phytoplankton, and are themselves the
greatest planktonic contribution to the
food of fishes. Most of them are herb-
ivorous. Two groups, the cladocera
and the copepods are most conspicuous.
2 Cladocera (Subclass Branchiopoda,
Order Cladocera) or Water Fleas
a Life History
1) During most of the year, eggs
which will develop without fertil-
ization (parthenogenetic) are
deposited by the female in a dorsal
brood chamber. Here they hatch
into minature adults which escape
and swim away.
2) As unfavorable conditions develop,
males appear, and thick-walled
sexual eggs are enclosed in egg
cases called ephippia which can
often endure freezing and drying.
3) Sexual reproduction may occur
at different seasons in different
species.
4) Individuals of a great range of
sizes', and even ephippia, are
thus encountered in the plankton,
but there is no "larval" form.
b Seasonal variation - Considerable
variation may occur between winter
and summer forms of the same
species in some cases. Similar
variation also occurs between arctic
and tropical situations.
c Forms commonly encountered as
open water plankton include:
Bosmina longirostris and others
Daphnia galeata and others
Other less common genera are:
Diaphanosoma, Chydorus, Sida,
Acroperus, Cerlodapnnia, Bytho-
trephes, and the carnivorous
Leptodora and Polyphemus.
d Heavy blooms -of Cladocerans may
build up in eutrophic waters.
The copepods (order Copepoda) are the
perennial microcrustacea of open waters,
both fresh and marine. They are the
most ubiquitous of animal plankton.
a Cyclops is the genus most often
found by the National Water Quality
Sampling Network activities. Eucy-
clops, Paracyclops, Diaptomus,
Canth'ocamptus,
J-.imnocalanus~are other forms
reported to be planktonic.
Copepods hatch into a minute char-
acteristic larvae called a nauplius
which differs considerably from the
adults. After five or six moults, the
copepodid stage is reached, and after
six more moults, the adult. These
larval stages are often encountered
and are difficult to identify.
II 12-2
-------
Animal Plankton
B Class Insecta
1 Only a single species of insect can be
ranked as a true plankton, this is the
midge fly Chaborus (approx. 8 spp,
formerly Corethra).
2 The larva of this insect has hydrostatic
organs that enable it to remain perman-
ently suspended in the water.
3 It is usually found in the depths during
the daytime, but comes to the surface
at night.
V OCCASIONAL PLANKTERS
A While the protozoa, rotifers, and micro-
crustacea make up the bulk of the plankton,
there are many other groups as mentioned
above that may also occur. Locally or
periodically these may be of major import-
ance. Examples are given below.
B Phylum Coelenterata
1 Polyps of the genus Hydra may become
detached and float about hanging from
the surface film or floating detritus.
2 The freshwater medusa Craspedacusta
occasionally appears in lakes or reser-
voirs in great numbers.
C Phylum Platyhelminthes
1 Minute Turbellaria (relatives of the
well known Planaria) are sometimes
taken with the plankton in eutrophic
conditions. They are readily confused
with ciliate protozoa.
2 Cercaria larvae of Trematodes (flukes)
parasitic on certain wild animals,
frequently appear in great numbers.
When trapped in the droplets of water
on a swimmer's skin, they attempt to
bore in. Man not being their natural
host, they fail. The resultant irritation
is called "swimmer's itch". Some can
be identified, but many unidentifiable
species may be found.
3 In many areas of the world, cercaria
larvae of human parasites such as the
blood fluke Schistosoma japonicum may
live as plankton, and penetrate the human
skin directly on contact.
D Phylum Nemathelminth.es
1 Nematodes (or nemas) or roundworms
approach the bacteria and the blue-green
algae in ubiquity. They are found in
the soil and in the water, and in the air
as dust. In both marine and fresh waters
and from the Arctic to the tropics.
2 Although the majority are free living,
some occur as parasites of plants,
animals, and man, and some of these
parasites are among out most serious.
3 With this distribution, it is obvious that
they will occasionally be encountered as
plankton. A more complete discussion
of nematodes and their public health
implications in water supplies will be
found elsewhere (Chang, S L.).
E Additional crustacean groups sporadically
met with in the plankton include the following:
1 Order Anostraca or fairy shrimps
(formerly included with the two
following orders in the Euphyllopoda)
primarily planktonic in nature.
a Extremely local and sporadic, but
when present, may be dominating.
b Artemia, the brine shrimp, can
tolerate very high salinities.
c Very widely distributed, poorly
> understood.
2 Order Notostraca, the tadpole shrimps.
Essentially southern and western in
distribution.
3 Order Conchostraca, the clam shrimps.
Widely distributed, sporadic in occur-
rence. Many local species.
4 Subclass Ostracoda. the seed shrimps.
Up to 3 in. in length. Essentially
benthic but certain species of Cypris.
and Notodromas may occur in consid-
erable numbers as plankton at certain
times of the year.
5 Certain members of the large subclass
Malacostraca are limnetic, and thus,
planktonic to some extent.
a The scuds, (order Amphipoda) are
essentially benthic but are sometimes
collected in plankton samples around
II 12-3
-------
Animal Plankton
weed beds or near shore. Nekto-
planktonic forms include Pontoporela
and some species of Gammarus.
b The mysid, or opossum shrimps are
represented among the plankton by
Mysis relicta, which occurs in the
deeper waters, large lakes as far
north as the Arctic Ocean.
F The Class Archnoidea, Order Hydracarina
(or Acari) the mites. Frequent in plankton
tows near shore although Unionicola crass-
ipes has been reported to be virtually
planktonic.
G The phylum Mollusca is but scantily
represented in the freshwater plankton,
in contrast to the marine situation.
Glochidia (ciliated) larvae are occasion-
ally collected, and snails now and then
glide out on a quiet surface film and are
taken in a plankton net. An exotic
bivalve Corbicula has a planktotrophic
veliger stage.
H Eggs and other reproductive structures
of many forms including fish, insects, and
rotifers may be found in plankton samples.
Special reproductive structures such as
the statoblasts of bryozoa and sponges,
and the ephippia of cladocerans may also
be included.
I Adventitious and Accidental Plankters
Many shallow water benthic organisms
may become accidentally and temporarily
incorporated into the plankton. Many of
those in the preceding section might be
listed here, in addition to such forms as
certain free living nematodes, small
oligochaetes, and tardigrades, Collembola
and other surface film livers are also
taken at times but should not be mistaken
for plankton. Fragments and molt skins
from a variety of arthropods are usually
observed.
Pollen from terrestrial or aquatic plants
is often unrecognized, or confused with
one of the above. Leaf hairs from
terrestrial plants are also confusing to
the uninitiated, they are sometimes
mistaken for fungi or other organisms
(and vice versa).
In flowing waters, normally benthic
(bottom living) organisms are often found
drifting freely in the stream. This
phenomenon may be constant or periodic.
When included in plankton collections,
they must be reported, but recognized
for what they are.
REFERENCES
1 Edmondson, W. F., ed. Ward and
Whipples's Freshwater Biology, 2nd
Edition, Wiley & Sons, Inc., New York.
1959.
2 Hutchinson, G. Evelyn. A Treatise on
Limnology. Vol. 2. Introduction to
Lake Biology and the Limnoplankton.
Wiley. 1115 pp. 1967.
3 Lackey, J. B. Quality and Quantity of
Plankton in the South End of Lake
Michigan in 1952. JAWWA.
36:669-74. 1944.
4 McGauhey, P.H., Eich, H.F., Jackson,
H.W., and Henderson, C. A Study
of the Stream Pollution Problem in the
Roanoke, Virginia, Metropolitan
District. Virginia Polytech. Inst.,
Engr. Expt. Sta.
5 Needham, J. G. and Lloyd, J. T. The
Life of Inland Waters. Ithaca, New
York, Comstock Publishing Co., Inc
. 1937.
6 Newell, G. E. and Newell, R. C.
Marine Plankton. Hutchinson Educ.
Ltd. London. 221 pp. 1963.
7 Palmer, C.M. and Ingram, W.M.
Suggested Classification of Algae and
Protozoa in Sanitary Science.
Sew. & Ind. Wastes. 27:1183-88.
1955.
II 12-4
-------
Animal Plankton
8 Pennak, R.W. Freshwater Invertebrates 10 Welch, P.S. Limnology, McGraw-Hill
of the United States. The Ronald Press, Book Co., Inc., New York. 1935.
New York. 1953.
9 Sverdrup, H.W., Johnson, M.W., and
Fleming, R.H. The Oceans, Their
Physics, Chemistry and General This outline was prepared by H.W. Jackson,
Biology. Prentice-Hall, Inc., New York. Chief Biologist, National Training Center,
1942. FWPCA, Cincinnati, OH 45226.
II 12-5
-------
Animal Plankton
3/4
Phylum PROTOZOA
Free Living Representatives
I. Flagellated Protozoa, Class Mastigophora
Anthophysis
Pollution to Her ant
Pollution tollerant
19/1
Colony of Poteriodendron
Pollution tollerant, 35/1
II. Ameboid Protozoa, Class Saroodina
Dimastigamoetoa
Pollution tollerant
10-50/i
Huelearia.reported
to be intollerant of
pollution, 45/i-
III. Ciliated Protozoa, Class Ciliophora
Colpoda
Pollution tollerant
20-120 jai
Holophrya,reported
to be intollerant of
pollution, 35/i
Difflueia
Pollution tollerant
60-500/4
Bpistylis. pollution
tollerant. Colonies often
maerosoopio.
H.W.Jaokson
-------
Animal Plankton
PLANKTONIC PROTOZOA
Peranema trichophorum
Top
Side
Chaos
Arcella
vulgaris
Actinosphaerium
»
&
, wrcii.cJii
111
**5fe'
Vorticella
Codonella
cratera
Tintinnidium
fluviatile
-------
Animal Plankton
PLANKTONIC ROTIFERS
Various Forms of Keratella cochlearis
Synchaeta
pectinata
Polygarthra
vulgaris
Brachionus
quadridentata
Rotaria sp
-------
Animal Plankton
SOME PLANKTONIC CRUSTACEANS
CRUSTACEANS
Copepod; Cyclops. Order Copepoda
2-3 mm
Water Flea;
Daphnia
A Nauplius larva of a Copepod
1-5 mm
Order Cladocera
2-3 mm
OSTRACODE
Left: Shell closed Right: Appendages extended
1-2 mm
II 12-9
-------
Animal Plankton
PLANKTONIC ARTHROPODA
A mysid shrimp - crustacean
A water mite - arachnid
Chaoborus midge larva - Insect
Aspects in the life cycle of the human tapeworm
Diphyllobothrium la turn, class Cestoda. A. adult as in human
intestine; B. procercoid larva in copepod; C. plerocercoid
larva in flesh of pickerel (X-ray view).
H.W. Jackson
II 12-10
-------
LABORATORY EXERCISES
General Laboratory Instructions 15
Types of Algae 16
Identification of Diatoms 17
Identification of Animal Plankton 18
-------
GENERAL LABORATORY INSTRUCTIONS
I GROUND RULES
A Students will be assigned to specific
benches or seats.
B Certain items of equipment such as
microscopes or other instruments
will be assigned for the use of specific
students or groups.
C Certain reference books and other items
will be assigned to a "Central Stores"
table from which they may be borrowed,
and to which they should be returned
when not in use. The person at whose
desk they are located at the end of the
period is responsible for returning them
to Central Stores.
II FLASH CARDS
A A major objective of certain laboratory
sessions is to become familiar with a
given group or groups of organisms.
This means associating names with
organisms and/or parts. One of the
most effective devices to accomplish
this objective has been found to be the
individually prepared "flash card. "
This consists of a card (3X5. for ex-
ample) with a sketch of an organism on
one side, and its classification written
on the other side. For later study, a
pack of such cards is arranged with all
sketches up, for example. Leaf through
the pack. Your sketch will remind you
of the organism you observed in labora-
tory or elsewhere. Think its name. If
you are in doubt, turn the card over to
check yourself. When you have learned
to recognize all the sketches in the pack
in hand, turn it over and go over the
names, recalling in your mind the
organisms (sketch) each refers to.
B A few sample flash cards will be fur-
nished. You are herby instructed to
prepare additional cards for yourself
of as many different organisms as
BI. MET. lab. la. 4. 69
possible in every laboratory or field
session where it is appropriate. The
instructors will be glad to check your
identification or other items on request
but cannot take it upon themselves to
call in or check all cards. Make fre-
quent references to the "Finder. " A
supply of blank cards will be maintained
at Central Stores.
C Identification represents an expenditure
of time and effort. If this achievement
Is retained on a 3X5 card, then repeti-
tious time in identification may be saved
later. Partial identification or even no
identification can be completed later,
as one's competence and resource aids
are enlarged. Code letters or numbers
can be used on these unknowns and when
identification becomes more complete,
data may be more meaningful. Date
collected, location, and selected known
environmental parameters can enhance
the usefulness of such a set, if these
are added to each card (see Figure 1).
Ill LABORATORY SESSIONS
The laboratory sessions will not all be con-
ducted in exactly the same way, as several
different instructors will be involved. Where
no special procedure is indicated at the end
of the lecture outline covering a given group,
the instructor will give verbal instructions
on the spot.
IV PRINCIPA L OBJECTIVES OF THE FIRST
LABORATORY SESSION WILL BE:
A Assignment of Places and Equipment
B Use of the Microscope
C Use of a Biological Key
These laboratory instructions were prepared
by H. W. Jackson, Chief Biologist, and
Ralph M. Sinclair, Aquatic Biologist.
National Training Center, FWPCA,
Cincinnati, OH 45226.
II 15-1
-------
General Laboratory Instructions
Figure 1
-------
Cyanophyta
fr
Hormogonales
Lyngbya
Species of this genus have been found to occur
in surface waters, attached to substrates and
in some cases associated with pollution. The
filament of cells is enclosed by a sheath. Cells
range from 4 to 20 microns in diameter and 4 to
10 microns in length.
P. Arthropoda
C. Ins e eta
O. Ephemeroptera (Mayflies)
Mayflies are common inhabitants of both lakes and
streams. They are gill breathers which are usually
found crawling about on rocks. Most genera can be
distinguished from the stoneflies by the presence of
3 caudal filaments.
P. Rotatoria (Rotifers)
C. Monogononta
F. Brachionidae
G. Keratella
The genus Keratella represents one of the forms of
rotifers which are often found in the plankton.
suoporujsui
-------
LABORATORY EXERCISES ON TYPES OF ALGAE
I OBJECTIVES
A To learn the terminology and techniques
for identifying the major groups of algae.
B To learn to recognize the more common
genera on sight.
B A representative sample of the algal group
under consideration will be distributed.
Make a wet mount and focus at lOOx on a
portion of the algal specimen.
C Using the algae key in your manual follow
through the identification with the in-
structor, making notes and drawings of
important characteristics.
II MATERIALS
A Compound microscopes equipped with
10.20,40, and lOOx objectives and
mechanical stages.
B Microscope lamps, glass slides, cover
glasses, pipettes, algal samples, iden-
tification keys, and supplemental books.
Ill PROCEDURE
D A discussion will accompany and follow
the identification pointing out the most
important characters and how they were
used in the classification.
This same procedure will be followed
with several other algal specimens
which illustrate the major morphological
forms within a certain group.
F Unknown samples will be distributed for
identification by the class.
A A short preview of the characteristics of
the specific group will be presented.
A discussion of the classification and
importance of the specimen will follow
each identification.
This outline was prepared by M.E.
Bender, Former Biologist, FWPCA
Training Activities, SEC.
BI. MIC. cla. lab. 11.4.70
II 16-1
-------
LABORATORY: IDENTIFICATION OF DIATOMS
I OBJECTIVES
A To become familiar with important
structural features of diatoms.
B To learn to recognize some common forms
at sight.
C To learn to identify less common forms
using technical keys.
II PROCEDURE
A Transfer a drop of the water sample con-
taining diatoms to a microscope slide. Cover
with cover glass and observe under low
power (10X) of microscope.
1 Do all of the diatom cells appear to
have the same shape? Do some have
square ends and some rounded ends ?
Touch the cover glass with your pencil
several times as you observe through
the microscope and note the relationship
of the two types of ends to one another.
2 Find a place where a round-ended and a
square-ended cell are close together
and observe these under oil power
(90X). The round-ended view is that of
the top or bottom of the diatom cell and
is called the "valve" view. The square
ended view is that of the side of the cell
and is called the "girdle" view.
3 In the valve view note the cross lines in
the wall. In this diatom there are many
fine lines and a smaller number of
coarse lines. The former are present
in all diatoms and are called "striae" or
"striations. " The latter are present
only in some genera of diatoms and are
called "septa, " or in other genera,
"costae. " Which of the two types of
lines are continuous from side to side ?
The space left in the center by the in-
terrupted lines is known as a "false
(pseudo) raphe. "
What is the predominant color of the
diatom? How many plastids ? In
diatoms, the identification is based al-
most entirely on the characteristics of
the cell wall.
Make an outline drawing, at least 3
inches long, of a valve view and a girdle
view of the diatom. Show the markings
in the upper third of each. Label the
striae, septa, and false-raphe. Make
a drawing of what you image an end
view or cross(transverse) section view
would be like.
Using the key, identify your specimen,
listing the alternatives selected.
B Use the key to identify other unknowns as
far as possible, listing the alternatives
selected in the key. Make a sketch of
Navicula and Cyclotella if you identify these
forms.
Ill IMPORTANT TERMS
Capitate - having a known-like end.
Costae - coarse transverse ribs in wall.
False raphe - (see pseudoraphe)
Frustule - the wall of the diatom.
Girdle view - the side view, in which the
diatom appears to have square or blunt ends.
Nodule - a lump-like swelling in the center or
ends of the valve.
Pseudoraphe - a clear space extending the
length of the diatom and bordered on both
sides by striae.
Punctae - the dots which comprise the
striae.
BI. MIC. cla. lab.
II 17-1
-------
Laboratory: Identification of Diatoms
Raphe - a longitudinal line (cleft) bordered on
both sides of striae.
Septa - a self-like partition in the diatom,
appearing often as a coarse line.
Striae - fine transverse lines especially
evident in the valve view.
Valve view - the top or bottom view, in which
the diatom has rounded ends, or is circular
in outline.
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 I (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 Experi-
mental 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 Adjucant Regions,
Farlowia. 2:143-221. 1945.
7 Smith, G. M. Class Bacillariophyceae.
Freshwater Algae of the United States,
McGraw-Hill Book Co. New York.
pp 440-510 2nd Ed. 1950.
8 Tiffany, L.H., and Bntton, M.E. Class
Bacillariophyceae. The Algae of
Illinois, University of Chicago Press.
pp 214-296. 1952.
9 Ward, H.B., and Whipple, G.C. Class I,
Bacillariaceae (Diatoms). Freshwater
Biology, J. Wiley & Sons. New York.
pp 171-189. 1948.
10 Weber, C.I. A Guide to the Common
Diatoms at Water Pollution Surveillance
System Stations. USDI. FWPCA,
Cincinnati, OH. 1966.
11 Whipple, G.C., Fair, G. M., and Whipple.
M.C. Diatomaceae. Microscopy of
Drinking Water. Chapter 21, 4th ed.
J. Wiley and Sons, New York. 1948.
This outline was prepared by L. G. Williams,
Aquatic Biologist, Formerly with Research
and Development, Cincinnati Water Research
Laboratory, FWPCA, SEC.
II 17-2
-------
LABORATORY: IDENTIFICATION OF ANIMAL PLANKTON
I INTRODUCTION
A The great majority of organisms commonly
encountered in plankton analysis work are
plants or at least plant-like (holophytic).
From time to time, however, animals
appear (holozoic or nonchlorophyll bearing
forms), and the ability to recognize them
may be quite important.
B Many animals are soft bodied and so are
best observed in the living condition, as
they shrink and become otherwise distorted
on preservation. There are consequently
many which will not be available in a
suitable form for the following exercise.
Only such forms will be dealt with as can
readily be obtained alive, or which retain
essential characteristics on preservation.
Examine your specimen carefully, '
then read the first couplet of
statements in the key (la and Ib).
Since the specimen is large enough
to see, it obviously could not be the
object of statement la. Therefore
due to the nature of the key (as
explained in the second paragraph of
the introduction) the second alternative
(Ib) must apply. This alternative
instructs us to proceed to couplet 2.
From here on, follow from couplet
to couplet, considering each couplet
by itself, until a final selection leads
to a name. If this name or couplet
is, followed by another couplet
number, this means that the group
named is further subdivided.
II OBJECTIVES
A To Study the Nature and Use of a Key for
Identifying Organisms
B To Introduce the Beginners to the Use of
the Microscope
C To Learn to Recognize Basic Animal Types
D To Identify Animal Plankton Species as
Available, and to Become Familiar with
the Literature
III PROCEDURE
A The Use of the Biological Key
1 Obtain a "Basic Invertebrate Collection"
from the instructor.
2 Select a specimen designated by the
instructor, and turn to the "Key to
Selected Larger Groups of Aquatic
Animals "
Identify the other specimens in the
Basic Invertebrate Collection in the
same way.
Carry the identification further, to
genus and species if possible, in one
or more of the more detailed keys
listed at the end of the "Key to Selected
Larger Groups of Aquatic Animals. "
B The Use of the Microscope
1 Obtain preliminary information from
the instructor as to how to set up and
operate the instrument.
2 Place a prepared slide of a printed letter
on the stage and observe it successively
under low (100X) and high (45X) powers.
When the letter is right side up to you,
how does it appear through the microscope?
3 Place a prepared slide of a micro -
crustacean on the stage and identify it
using the "Key to Selected Larger Groups
of Aquatic Animals. " Continue your
BL MIC. cla. lab. 5b. 10. 66
II 18-1
-------
Laboratory: Identification of Animal Plankton
identification as far as possible using
Eddy and Hodson's "Taxonomic Keys. "
Prepare a "wet mount" under the
direction of the instructor and identify
the organism. Confirm your identifica-
tion in one or more of the technical
reference books available.
Identify each of the specimens in the
reference collection (labeled "III C")
as to phylum and class, and then genus
and species if possible (do not spend
undue time on the species without
assistance).
1 Make a sketch of at least one organism
of each phylum observed as an example
of a type.
Review the collection of "Explano-
mounts. " Sketch and study any with
which you are not familiar.
Work each one through the keys as if
you did not know its name.
D Examine the living material provided.
Sketch and identify animal forms
encountered as far as time permits. Can
you draw any conclusions as to the types
of animal life found in the various
habitats indicated?
This outline was prepared by H. W. Jackson,
Chief Biologist, National Training Center,
FWPCA, Cincinnati, OH 45226.
II 18-2
-------
CHAPTER El
TECHNIQUES OF PLANKTON METHODOLOGY
Techniques of Plankton Sampling Programs
Preparation and Enumeration of Plankton in the Laboratory
Calibration and Use of Plankton Counting Equipment
Preparation of Permanent Diatom Mounts
Determination of Odors
Collection and Interpretation of Biological Lake Data
Determination of Plankton Productivity
Methods of Measuring Standing Crops of Plankton
Aerial Reconnaissance in Pollution Surveillance
1
2
3
4
5
6
8
9
10
-------
TECHNIQUES OF PLANKTON SAMPLING PROGRAMS
I INTRODUCTION
A A planned program of plankton analysis
should involve periodic sampling at weekly
intervals or more often.
1 Most interference organisms are small,
and hence have relatively short-life
histories.
2 Populations of such organisms may
fluctuate rapidly in response to chang-
ing water, weather, or seasons..
3 Seasonal growth patterns of plankton
tend to repeat themselves from year to
year, thus they are relatively predictable.
B A well-planned study or analysis of the
growth pattern ot plankton in one year will
provide a basis for predicting conditions
the following year.
1 Since the seasons and the years differ,
the more records are accumulated, the
more useful can they become.
2 As the time for an anticipated bloom of
some trouble maker approaches, the
frequency of analysis may be increased.
C Detection of a bloom in its early stages
will facilitate more economical control.
II FIELD ASPECTS OF THE ANALYSIS
PROGRAM
A Two general aspects of sampling are com-
monly recognized: quantitative and
qualitative.
1 Qualitative examination tells what is
present.
2 Quantitative tells how much.
3 Either approach is useful, a combination
if best.
B Equipment of collecting samples in the
field is varied.
1 A half-liter bottle will suffice for sur-
face samples of phytoplankton if carefully
taken. If zooplankton also are of interest,
2 or more liters should be collected.
(See below).
2 Plankton nets concentrate the sample
in the act of collecting, and capture
certain larger forms which escape from
the bottles. Only the more elaborate
types are quantitative however.
3 A kemmerer-type sampler is suggested
for depth samples.
4 Other methods such as the Clark-Bumpus
sampler or the Juday plankton trap may
be employed for special purposes.
C The location of sampling points is
important
1 Both shallow and deep samples are
suggested.
a "Shallow" samples should be taken
at a depth of 6 inches to one foot.
b "Deep" samples should be taken at
such intervals as the depth of the
reservoir permits. There should be
at least one open water sampling
point.
c Each major bay or shoal area should
have at least one sampling point.
d Additional sampling stations should
be established on the basis of ex-
perience and resources.
e Samples may be composited if nec-
essary to give an overall summary of
conditions. Such summaries are not
advised and should be interpreted
with care.
BI. MIC. enu.9g. 3.70
III 1-1
-------
Techniques of Plankton Sampling Programs
A standardized vertical haul however,
can be useful for routine comparisons.
Ill RECORDS
Field conditions greatly affect the plankton,
and a record thereof should be carefully
identified with the collection.
A Time of day, turbidity of the water, and
relative cloudiness affect the amount of
light, which affects the vertical distribu-
tion of many forms.
B Water temperature affects growth rate and
behavior.
C Wind causes water drift, and wave action
breaks up colonial forms and disperses
accumulations.
D Other conditions may be usefully recorded
if circumstances permit.
IV FIELD PRESERVATION OF SAMPLES
Provision should be made for the field
stabilization of the sample until the laboratory
examination can be made. Techniques and
materials are listed below. No "ideal" pre-
servative or technique has yet been developed,
each has its virtues.
A Refrigeration or icing. The container
containing the sample can be cooled, but
under no circumstances should ice be
dropped into the sample.
B Preservation by 3-5% formaline is time-
tested and widely used. Formaline shrinks
animal tissue, fades colors, and makes all
forms brittle.
C Ultra-violet sterilization is useful in the
laboratory to retard decomposition of
plankton.
D Lugol's solution is sometimes useful.
E A special thimerosal preservative was
developed by the FWPCA Water Pollution
Surveillance System which has proved very
satisfactory and is described in reference
No. 2.
V SUMMARY AND CONCLUSIONS
A The field sampling program should be
carefully planned to evaluate all signifi-
cant locations in the reservoir or stream,
giving due consideration to the capacity
of the laboratory.
B Adequate records and notes should be
made of field conditions and associated
with the laboratory analyses in a permanent
file.
C Once a procedure for processing plankton
is adopted, it should be used exclusively
by all workers at the plant.
D Such a procedure should enable the water
plant operator to prevent plankton troubles
or at least to anticipate them and have
corrective materials or equipment
stockpiled.
REFERENCES
1 Hutcheson, George E. A Treatise on
Limnology. John Wiley and Co. New
York. 1957.
2 Jackson, H. W. Biological Examination (of
plankton) Part III in Simplified Procedures
for Water Examination. AWWA Manual
M12. Am. Water Works Assoc., N. Y.
1964.
3 Lackey, J. B. The Manipulation and Count-
ing of River Plankton and Changes in
Some Organisms Due to Formalin Pre-
servation. Public Health Reports, 53:
2080-93. 1938.
Ill 1-2
-------
Techniques of Plankton Sampling Programs
4 Olson, Theodore A. and Burgess,
Frederick J. Pollution and Marine
Ecology. Interscience Publishers.
364pp. 1967.
5 Palmer, C.M. Algae in Water Supplies.
U. S. Department of Health, Education,
and Welfare, Public Health Service
Publication No. 657, Superintendent of
Documents, Washington 25, D. C.
6 Weber, C. I. Methods of Collection and
Analysis of Plankton and Periphyton
Sda Samples in the Water Pollution
Surveillance System. App. and
Devel. Rep. (AQC Lab., 1014 Broadway,
Cincinnati, OH 45202) 19 pp. 1966.
7 Welch, P.S. Limnological Methods.
The Blakiston Co., Phila. Toronto.
1948.
8 Williams, L. G. Plankton Population
Dynamics, in National Water Quality
Network, Supplement 2, U. S. PHS
Pub. No. 663. 1962.
This outline was prepared by H. W. Jackson,
Chief Biologist, National Training Center,
FWPCA, Cincinnati, OH 45226.
in 1-3
-------
PREPARATION AND ENUMERATION OF PLANKTON IN THE LABORATORY
I RECEPTION AND PREPARATION OF
SAMPLES
A Preliminary sampling and analysis is an
essential preliminary to the establishment
of a permanent or semi-permanent program.
B Concentration or sedimentation of preserved
samples may precede analysis
1 Batch centrifuge
2 Continuous centrifuge
3 Sedimentation
C Unpreserved (living) samples should be
analyzed at once or refrigerated for future
analysis.
II PREPARATION OF THIMEROSAL
PRESERVATIVE
A The Water Pollution Surveillance System
of the FWPCA has developed a modified
thimerosal preservative. (Williams, 1962;
Weber. 1967) Sufficient stock to make an
approximately 3. 5% solution in the bottle
when filled is placed in the sample bottle
in the laboratory. The bottle is then filled
with water in the field and returned to the
laboratory for analysis.
B Preparation of Thimerosal Preservative
1 Thimerosal is available from many
chemical laboratory supply houses;
one should specify the water soluble
sodium salt.
2 Thimerosal stock: dissolve approximately
1 gram of sodium borate (borax) and
approximately 1 gram of thimerosal in
1 liter of distilled water.
The amount of sodium borate and
thimerosal may be varied slightly to
adjust to different waters, climates,
and organic contents.
3 Prepare a saturated aqueous Lugol's
solution as follows:
a Add 60 grams of potassium iodide (KI)
and 40 grams of iodine to 1 liter of
distilled water.
Prepare the preservative solution by
adding approximately 1.0 ml of the
Lugol's solution to 1 liter of
thimerosal stock.
Ill SAMPLE ANALYSIS
Microscopic examination is most frequently
employed in the laboratory to determine
what plankton organisms are present and
how many there are.
1 Optical equipment need not be elaborate
but should include.
a Compound microscope with the
following equipment:
1) Mechanical stage
2) Ocular 10X, with Whipple type
counting eyepiece or reticule
3)
Objectives
approx.
approx.
approx
approx.
10X(16 mm)
20X(8 mm)
40X(4 mm)
95X(1 8 mm)(optional)
BI.MIC.enu.l5e.6.68
III 2-1
-------
Preparation and Enumeration of Plankton
A 4OX objective with a working
distance of 12. 8 mm and an erect
image may be obtained as special
equipment. A water immersion
objective (in addition to oil) might
be considered for use with water
mounts.
Binocular eyepieces are optional
Stage micrometer (this may be
borrowed, if necessary, as it is
usually used only once, when the
equipment is calibrated)
b Inverted microscopes offer certain
advantages but are not widely available.
The same is true of some of the
newer optical systems such as phase
contrast microscopy. These are
often excellent but
expensive for routine plant use.
Precision made counting chambers
are required for quantitative work with
liquid mounts.
a Sedgwick-Rafter cells (hereafter
referred to as S-R cell) are used for
routine counts of medium and larger
forms.
b Extremely small forms or "nanno-
plankton" may be counted by use of
the nannoplankton (or Palmer) cell,
a Fisher -Littman cell, a hemacytometer,
the Lackey drop method, or by use of
an inverted microscope.
Previous to starting serious analytical
work, the microscope should be
calibrated as described elsewhere
Dimensions of the S-R cell should also
be checked, especially the depth
Automatic particle counters may be
useful for coccoid organisms.
B Quantitative Plankton Counts
1 All quantitative counting techniques
involve the filling of a standard cell of
known dimensions with either straight
sample or a concentrate or dilution
thereof.
The organisms in a predetermined
number of microscope fields or other
known area are then observed, and by
means of a suitable series of multiplier
factors, projected to a number or
quantity per ml gallon, etc.
Direct counting of the unconcentrated
sample eliminates manipulation, saves
time, and reduces error If frequency
of organisms is low, more area may
need to be examined or concentration of
the sample may be in order.
Conventional techniques employing
concentration of the sample provide more
organisms for observation, but because
they involve more manipulations,
introduce additional errors and take
more time
C Several methods of counting plankton are
in general use.
1 The numerical or clump count is
regarded as the simplest
a Every organism observed must be
enumerated. If it cannot be identified,
assign a symbol or number and make
a sketch of it on the back of the
record sheet.
b Filaments, colonies and other
associations of cells are counted
as units, equal to single isolated
cells Their identity as indicated
on the record sheet is the key to the
significance of such a count
2 Individual cell count In this method,
every cell of every colony or clump
of organisms is counted, as well as
each individual single-celled organism.
3 The areal standard unit method offers
certain technical advantages, but also
involves certain inherrent difficulties.
a An areal standard unit is 400 square
microns. This is the area of one
of the smallest subdivided squares
III 2-2
-------
Preparation and Enumeration of Plankton
in the center of the Whipple eyepiece
at a magnification of 100X
b In operation, the number of areal
units of each species is recorded on
the record sheet rather than the
number of individuals Average
areas of the common species are
sometimes printed on record sheets
for a particular plant to obviate the
necessity of estimating the area of
each cell observed individually
c The advantage of the method lies in
the cognizance taken of the relative
masses of the various species as
indicated by the area presented to
the viewer These areas, however, are
often very difficult to estimate.
4 The cubic standard unit method is a
logical extension of the areal method,
but has achieved less acceptance.
5 Separate field count
a In counting separate fields, the
question always arises as to how
to count organisms touching or
crossed by the edge of the Whipple
field Some workers estimate the
proportion of the organism lying
inside the field as compared to that
outside. Only those which are over
half way inside are counted.
b Another system is to select two
adjacent sides of the square for
reference, such as the top and left
boundaries. Organisms touching
these lines in any degree, from
outside or inside, are then counted,
while organisms touching the opposite
sides are ignored. It is important
to adopt some such system and
adhere to it consistently
c It is suggested that if separate
microscopic fields are examined,
a standard number of ten be adopted.
These should be evenly spaced in two
rows about one third of the distance
down from the top and one third of the
distance up from the bottom of the
S-R cell.
6 Multiple area count. This is an
extension of the separate field count
A considerable increase in accuracy
has recently been shown to accrue by
emptying and refilling the S-R cell,
after each group of fields are counted
and making up to 5 additional such
counts. This may not be practical with
high counts.
7 The strip count. When a rectangular
slide such as the S-R cell is used, a strip
(or strips) the entire length (or known
portion thereof) of the cell may be count-
ed instead of separate isolated fields.
Marking the bottom of the cell by evenly
spaced cross lines as explained elsewhere
greatly facilitates counting.
a When the count obtained is multiplied
by the ratio of the width of the strip
counted to the width of the cell, the
product is the estimated number of
organisms in the cell, or per ml
b When the material in the cell is
unconcentrated sample water, this
count represents the condition of
the water being evaluated without
further calculation
8 Once a procedure for concentration
and/or counting is adopted by a plant
or other organization it should be used
consistently from then on so that results
from year to year can be compared.
D Differential or qualitativej'counts" are
essentially lists of the kinds of organisms
found.
E Proportional or relative counts of special
groups are often very useful. For example,
diatoms. It is best to always count a stan-
dard number of cells.
F Plankton are sometimes measured by
means other than microscopic counts
1 Settled volume of killed plankton in an
Imhoff cone may be observed after a
standard length of time This will
evaluate primarily only the larger forms.
Ill 2-3
-------
Preparation and Enumeration of Plankton
A gravimetric method employs drying
at 60°c for 24 hours followed by ashing
at 600°C for 30 minutes. This is
particularly useful for chemical and
radiochemical analysis
A method for chlorophyll extraction
involves filtration, drying for 24 hours,
extraction with methyl alcohol, and
evaluation in a colorimeter or acetone
extracts, using a spectrophotometer
This is widely used in "productivity"
studies.
The membrane (molecular) filter has
a great potential, but a generally accept-
able technique has yet to be perfected.
B Adequate records and notes should be
made of field conditions and associated
with the laboratory analyses in a
permanent file.
C Optical equipment in the laboratory should
be calibrated.
D Once a procedure for processing plankton
is adopted, it should be used exclusively
by all workers at the plant
E Such a procedure should enable the water
plant operator to prevent plankton troubles
or at least to anticipate them and have
corrective materials or equipment
stockpiled.
a Bacteriological techniques for
cohform determination are widely
accepted.
b Nematodes and larger organisms can
readily be washed off of the membrane
after filtration
c It is also being used to measure
ultraplankton that pass treatment
plant operations.
d Membranes can be cleared and
organisms deposited thereon
observed directly, although accessory
staining is desirable.
e Difficulties include a predilection of
extremely fine membranes to clog
rapidly with silt or increase in
plankton counts, and the difficulty
of making observations on individual
cells when the organisms are piled
on top of each other It is sometimes
necessary to dilute a sample to obtain
suitable distribution
IV SUMMARY AND CONCLUSIONS
The field sampling program should be
carefully planned to evaluate all significant
locations in the reservoir or stream, giving
due consideration to the capacity of the
laboratory
REFERENCES
1 Ely Lilly Company. Merthiolate As a
Preservative Ely Lilly & Co
Indianapolis 6, Indiana.
2 Gardiner, A C. Measurement of
Phytoplankton Population by the
Pigment Extraction Method Jour.
Marine Biol. Assoc 25_(4):739-744. 1943.
3 Goldberg, E D , Baker, M , and Fox, D. L.
Microfiltration m Oceanographic
Research Sears Foundation. Jour.
Mar Res jj_- 194-204 1952
4 Ingram, W. M. , and Palmer, C M.
Simplified Procedures for Collecting,
Examining, and Recording Plankton in
Water. Jour. AWWA. 44(7).617-624.
1952.
5 Jackson, H. W. Biological Examination
(of plankton) Part III in Simplified Pro-
cedures for Water Examination. AWWA
Manual M 12. Am. Water Works Assoc.
N.Y. 1964
6 Lund, J W G. , and Tailing, J.F.
Botanical Limnological Methods with
Special References to the Algae.
Botanical Review. 23_(8&9)- 489-583.
October, 1957
III 2-4
-------
Preparation and Enumeration of Plankton
7 Weber, C.I. The Preservation of Plankton
Grab Samples. Water Pollution
Surveillance System, Applications and
Development Report No. 26, USDI,
FWPCA. Cincinnati, Ohio. (1967)
8 Williams, L. G. Plankton Population
Dynamics. National Water Quality
Network Supplement 2. U.S. Public
Health Service Publ. No. 663. (1962)
Wohlschlag, D.D., and Hasler, A.D.
Some Quantitative Aspects of Algal
Growth in Lake Mendota. Ecology.
32(4): 581-593. (1951)
This outline was prepared by H.W. Jackson.
Chief Biologist, National Training Center,
FWPCA, Cincinnati, OH 45226.
Ill 2-5
-------
CALIBRATION AND USE OF PLANKTON COUNTING EQUIPMENT
I INTRODUCTION
A With the exception of factory-set
instruments, no two microscopes can
be counted upon to provide exactly the
same magnification with any given com-
bination of oculars and objectives. For
accurate quantitative studies, it is there-
fore necessary to standardize or "calibrate"
each instrument against a known standard
scale. One scale frequently used is a
microscope slide on which two millimeters
are subdivided into tenths, and two addi-
tional tenths are subdivided into hundredths.
Figure 3.
B In order to provide an accurate measuring
device in the microscope, a Whippie
Plankton Counting Square or reticule '
(Figure 2a) is installed in one ocular
(there are many different types of reticules).
This square is theoretically of such a size
that with a 10X objective, a 10X ocular,
and a tube length of 160 mm, the image
of the square covers a square area on the
slide one mm on a slide. Since this
objective is rarely attained however, most
microscopes must be standardized or
"calibrated" as described below in order
to ascertain the actual size of the W hippie
Square as seen through the microscope
(hereinafter referred to as the "Whipple
field"). This process is schematically
represented in Figures 5 and 7. If the
Whipple eyepiece is to be used at more
than one magnification, it must be recali-
brated for each. A basic type of monocular
microscope is shown in Figure 1.
C Microscopes with two eyepieces (binocular)
are a convenience but not essential. Like
modern cars they are not only great
"performers, " but also complicated to
service or, in this instance, calibrate.
On some instruments, changing the inter-
pupillary distance also changes the tube
length, on others it does not. The "zoom"
feature on certain scopes is also essentially
a system for changing the tube length.
The resultant is that in addition to calibra-
tion at each combination of eyepiece and
objective, any other factor which may
affect magnification must also be considered.
In some instances this may mean setting up
a table of calibrations at a series of micro-
scope settings.
Another procedure is to select a value
for each of the variables involved (inter-
pupillary distance, zoom, etc.) and
calibrate the scope at that combination.
Then each time the scope is to be used for
quantitative work, re-set each variable to
the value selected. A separate multipli-
cation factor must be calculated for each
adjustment which changes the magnification
of the instrument.
Since the Whipple Square can be used to
measure both linear dimensions and
square areas, both should be recorded on
an appropriate form. A suggested format
is shown in Figure 6.
(Data written in are used as an illustration
and are not intended to apply to any
particular microscope. An unused form
is included as Figure 6-A.)
II THE CALIBRATION PROCEDURE
A Installing the Whipple Square or Reticule
To install the reticule in the ocular
(usually the right one on a binocular
microscope), carefully unscrew the upper
lens mounting and place the reticule on
the circular diaphragm or shelf which will
be found approximately half way down inside
(Figure 4). Replace the lens mounting and
observe the markings on the reticule. If
they are not in sharp focus, remove and
turn the reticule over.
On reticules with the markings etched on
one side of a glass disc, the etched sur-
face can usually be recognized by shining
the disc at the proper angle in a light.
The markings will usually be in the best
focus with the etched surface down. If the
markings are sandwiched between two glass
discs cemented together, both sides are
alike, and the focus may not be quite as
sharp.
B Observation of the Stage Micrometer
Replace the ocular in the microscope and
observe the stage micrometer as is illus-
trated schematically in Figure 5: Calibration
of the Whipple Square. On a suitably ruled
form such as the one illustrated. Figure 6,
Calibration Data, record the actual distance
in millimeters subtended by the image of
BI. MET.mic. Id. 4. 70
in 3-1
-------
Calibration and Use of Plankton Counting Equipment
the entire Whipple field and also by each
of its subdivisions. This should be
determined for each significant settling of
the interpupillary distance for a binocular
microscope, and also for each combination
of lenses employed. Since oculars and
objectives marked with identical magnifi-
cation, and since microscope frames too
may differ, the serial or other identifying
number of those actually calibrated should
be recorded. It is thus apparent that the
determinations recorded will only be valid
when used with the lenses listed and on that
particular microscope.
C Use of the 20X Objective
Due to the short working distance beneath
a 46X (4mm) objective, it is impossible
to focus to the bottom of the Sedgewick-
Rafter plankton counting cell with this lens.
A 10X (16mm) lens on the other hand
"wastes" space between the front of the
lens and the coverglass, even when focused
on the bottom of the cell. In order to make
the most efficient use possible of this cell
then, an objective of intermediate focal
length is desirable. A lens with a focal
length of approximately 8 mm, having a
magnification of 20 or 2IX will meet these
requirements. Such lenses are available
from American manufacturers and are
recommended for this type of work.
E CHECKING THE CELL
The internal dimensions of a Sedgewick-Rafter
plankton counting cell should be 50 mm long
by 20 mm wide by 1 mm deep (Figure 8).
The actual horizontal dimensions of each new
cell should be checked with calipers, and the
depth of the cell checked at several points
around the edge using the vertical focusing
scale engraved on the fine adjustment knob of
most microscopes. One complete rotation of
the knob usually raises or lowers the objective
1 mm or 100 microns (and each single mark
equals 1 micron). Thus, approximately ten
turns of the fine adjustment knob should raise
the focus from the bottom of the cell to the
underside of a coverglass resting on the rim.
Make these measurements on an empty cell.
The use of a No. 1 or 1-1/2, 24 X 60 mm
coverglass is recommended rather than the
heavy coverglass that comes with the S-R
cell, as the thinner glass will somewhat con-
form to any irregularities of the cell rim
(hence, also making a tighter seal and reduc-
ing evaporation when in actual use). Do not
attempt to focus on the upper surface of the
rim of an empty cell for the above depth
measurements, as the coverglass is supported
by the highest points of the rim only, which
are very difficult to identify. Use the average
of all depth measurements as the "true" depth
of the cell. To simplify calculations below, it
will be assumed that we are dealing with a
cell with an average depth of exactly 1. 0 mm.
IV PROCEDURE FOR STRIP COUNTS USING
THE SEDGEWICK-RAFTER CELL
A Principles
Since the total area of the cell is 1000 mm2,
the total volume is 1000 mmj or 1 ml. A
"strip" the length of the cell thus constitutes
a volume (V,) 50 mm long, 1 mm deep, and
the width of the Whipple field.
q
The volume of such a strip in mm is:
Vj = 50 X width of field X depth
= SOXwX 1
= 50 w
In the example given below on the plate
entitled Calibration Data, at a magnification
of approximately 200X with an interpupillary
setting of "60", the width of the Whipple
field is recorded as approximately 0. 55 mm
(or 550 microns). In this case, the volume
of the strip is:
Vj = 50 w = 50 X 0. 55 = 27. 5 (mm )
B Calculation of Multiplier F actor
In order to convert plankton counts per
strip to counts per ml, it is simply
necessary to multiply the count obtained
by a factor (F,) which represents the
number of tinres the volume of the strip
examined (V}) would be contained in 1 ml or
1000 mm 3. Thus in the example given
above:
3
,-, volume of cell in mm
volume examined in mm
1000 1000
= = 2775
= 36.36
approx. 36
If more than one strip is to be counted,
the factor for two, three, etc., strips
could be calculated separately using the
same relationships outlined above, changing
only the measurement for the length of
III 3-2
-------
Calibration and Use of Plankton Counting Equipment
Figure 1. THE COMPOUND MICROSCOPE
A) coarse adjustment; B) fine adjustment;
C) arm or pillar; D) mechanical stage which
holds slides and is movable in two directions
by means of the two knobs; E) pivot or joint.
This should not be used or "broken" while
counting plankton; F) eyepiece (or ocular cf:
figure 4); G) draw tube. This will be found
on monocular microscopes only (those having
only one eyepiece). Adjustment of this tube
is very helpful in calibrating the microscope
for quantitative counting (Sec. 5. 5. 2. 2.).
H) body tube. In some makes of microscopes
this can be replaced with a body tube having
two eyepieces, thus making the 'scope into
a "binocular. " I) revolving nosepiece on
which the objectives are mounted; J) through
M are objectives, any one of which can be
turned toward the object being studied. In
this case J is a 40X, K is a 1()OX, L is a 20X,
and M is a 10X objective. The product of
the magnification power of the objective being
used times the magnification power of the
eyepiece gives the total magnification of the
microscope. Different makes of microscopes
employ objectives of slightly different powers,
but all are approximately equivalent. N) stage
of the microscope; O) Sedgwick-Rafter cell in
place for observation; P) substage condenser;
Q) mirror; R) base or stand; note: for
information on the optical system, consult
reference 3. (Photo by Don Moran.).
Ill 3-3
-------
Calibration and Use of Plankton Counting Equipment
Figure 2
Types of eyepiece micrometer discs or
reticules (reticules, graticules, etc.).
When dimensions are mentioned in the
following description, they refer to the
markings on the reticule discs and not to
the measurements subtended on the micro-
scope slide. The latter must be determined
by calibration procedures such as those
described elsewhere, (a) Whipple plankton
counting eyepiece. The fine rulings in the
subdivided square are sometimes extended to
the margin of the large square to facilitate
the estimation of sizes of organisms in
different parts of the field, (b) Quadrant
ruling with 8. 0 mm circle, for counting
bacteria in milk smears for example, (c)
Linear scale 5. 0 mm divided into tenths.
For measurement of linear dimensions.
(d) Porton reticule for estimating the size
of particles. The sizes of the series of discs
is based on the square root of two so that the
areas of successive discs double as they
progress in size.
Ill 3-4
-------
Calibration and Use of Plankton Counting Equipment
strip counted. Thus for two strips in the
example cited above:
V= 100W = 100 X 0.55 = 55 mm3
1000 _ 1000 _ 1n 9
F2= -V 18'2
It will however be noted that F = -j- .
Likewise a factor F_ for three strips
F,
would equal -*- or approximately 12, etc.
C An Empirical "Step-Off" Method
A simpler but more empirical procedure
for determining the factor is to consider
that if a strip 20 mm wide were to be
counted the length of the cell, that the
entire 1000 mnv* would be included since
the cell is 20 mm wide and 1 mm deep.
This 20 mm strip width can be equated to
1000 mm3. If a strip (or the total of 2 or
more strips) is less than 20 mm in width,
the quotient of 20 divided by this width will
be a multiplier factor for converting from
count per strip(s) to count per ml.
V
Thus in the example cited above where at
an approximate magnification of 200X and
with an interpupillary setting of 60, the
width of the Whipple field is . 55 mm. Then:
= 36.36 or approx. 36
(as above)
If two strips are counted:
andF2 =
= 18.2 = approx. 18,etc.
This same value could be obtained without
the use of a stage micrometer by carefully
moving the cell sidewise across the field
of vision by the use of a mechanical stage.
Count the number of Whipple fields in the
width of the cell. There should be approxi-
mately 18 in the instance cited above.
SEPARATE FIELD COUNT USING THE
SEDGEWICK-RAFTER CELL
A Circumstances of Use
The use of concentrated samples, local
established programs, or other circumstances
Figure 3. STAGE MICROMETER
The type illustrated has two millimeters divided into tenths, plus two additional
tenths subdivided into hundredths.
Micrometer Seal*
[.2 mm
Enlargement of Micrometer Scale
III 3-5
-------
Calibration and Use of Plankton Counting Equipment
Figure 4. Method of Mounting the Whipple Disc in an Ocular. Note the upper
lens of the ocular which has been car efully unscrewed, held in the left
hand, and the Whipple disc, held in the right hand. (Photo by
Don Moran).
HI 3-6
-------
Calibration and Use of Plankton Counting Equipment
CALIBRATION OK WHIPPLE SQUARE
as seen with 10X Ocular and 43X Objective
(approximately 430X total magnification)
Whipple Square as
seen through ocular
("Whipple field")
"Small quarei" subtend
one fifth of large squares:
.0052 mm or 5. 2w
"Large square" subtends
one lenth of entire Whipple
Square: . 026 nun or 26n
Apparent lines of sight
subtend . 26 mm or 260p
on stage micrometer
scale
PORTION OF MAGNIFIED IMAGE OF STAGE MICROMETER SCALE
Figure 5
CALIBRATION OF THE WHIPPLE SQUARE
The apparent relationship of the Whipple
Square is shown as it is viewed through a
microscope while looking at a stage
micrometer with a magnification of
approximately 43 OX (10X ocular and 43 X
objective).
Ill 3-7
-------
Calibration and Use of Plankton Counting Equipment
MICROSCOPE CALIBRATION DATA
Microscope No
79
Approximate
Mortification
Tube
Length, or
Interpupillary
Setting
Linear dimensions of Whipple
squaies in millimeters*
Whole
Large
Small
Factor for
Conversion
to count/ ml
100X. obtained with (2 S-R Stripe)
Objective
Serial No.
mil, 421/10$
and Ocular
Serial No
/J9t,7M.(/t>i\
50
1,0
10
/. 13 o
/.//S
/ ItO
0.113
O.lll
O I/O
OOJlt,
ejzm.
O. 03.A3L
s.q
9.0
200X, obtained with
(2 S-R Strips)
Objective
Serial No.
&/y«,f%?/y)
and Ocular
Serial No.
>5»
taO
7o
O.SLO
o.ss-o
o.su
O.05&,
O. OSS
0 QSV
O, O //£
001 10
0 O/09
/-71
/f.l
/f.A
400X, obtained with
(Nannoplankton)
(cell-20 fields )'
Objective
Serial No.
IM/Ai/A&x}
and Ocular
Serial No.
/i? 9^7*4 (fox)
.57?
(,0
7£>
O.S.&7
O.UZ
O.JteO
a.G3t>?
n.oa&S
n.oAUO
,0t>33
.0053
.0032
/73V.
17 f 6,.
/Ss-J.
*1 mm = 1000 microns
Microscope calibration data. The form
shown is suggested for the recording of
data pertaining to a particular microscope.
Headings could be modified to suit local
situations. For example, "Interpupillary
Setting" could be replaced by "Tube Length"
or the "2S-R Strips could be replaced by
"per field" or "per 10 fields. "
Figure 6
BI.AQ.pl. 8b. 7.66
HI 3-8
-------
Calibration and Use of Plankton Counting Equipment
MICROSCOPE CALIBRATION DATA
i
Microscope No.
Approximate
Magnification
Tube
Length, or
Interpupillary
Setting
Linear dimensions of Whipple
squares in millimeters*
Whole
Large
Small
Factor for
Conversion
to count/ ml
100X, obtained with (2 S-R Strips)
Objective
Serial No.
and Ocular
Serial No
200X, obtained with (2 S-R Strips)
Objective
Serial No.
and Ocular
Serial No.
400X, obtaine
Objective
Serial No.
and Ocular
Serial No.
(Nannoplankton)
d with (cell- 20 fields )
*lmm = 1000 microns
BI.AQ.pl 8. 10. 60.
Figure 6-A
MICROSCOPE CALIBRATION DATA
Suggested work sheet for the calibration of a microscope. Details will need to be adapted
to the particular instrument and situation.
in 3-9
-------
Calibration and Use of Plankton Counting Equipment
S-R COVER
GLASS 1
WATER IN 1MM
S-R CELL
ITMir ICKICCC C_
THICKNESS S-R SLIDE
Figure 7
A cube of water as seen through a Whipple square at IOOX magnification in
a Sedgewick-Rafter cell. The figure is drawn as if the microscope were
focused on the bottom of the cell, making visible only those organisms lying
on the bottom of the cell. The little "bug" (copepod) halfway up, and the
algae filament at the top would be out of focus. The focus must be moved up
and down in order to study (or count) the entire cube.
Ill 3-10
-------
Calibration and Use of Plankton Counting Equipment
may make it necessary to employ the more
conventional technique of counting one or
more separate Whipple fields instead of
the strip count method. The basic relation-
ships outlined above still hold, namely:
3
_ volume cell in mm
volume examined in mm*
B Principles Involved
The volume examined in this case will
consist of one or more squares the dimen-
sions of the Whipple field in area and 1 mm
in depth (Figure 7). Common practice
for routine work is to examine 10 fields,
but exceptionally high or low counts or
other circumstances may indicate that
some other number of fields should be
employed. In this case a "per field"
factor may be determined to be subsequently
divided by the number of fields examined
as with the strip count. The following
description however is based on an assumed
count of 10 fields.
C Calculation of Multiplier Factor
As stated above, the total volume
represented in the fields examined con-
sists of the total area of the Whipple fields
multiplied by the depth.
V4 = (side of Whipple field)2 X depth
(1 mm) X no. of fields counted)
For example, let us assume an approxi-
mate magnification of 100X (see Figures
6 and 7 and an interpupillary setting of
"50". The observed length of one side
of the Whipple field in this case is 1.13
mm. The calculation of V. is thus:
V4= side2 X depth X no. of fields
= 1.13 X 1.13 X 1 X10 = 12.8 mm3
The multiplier factor is obtained as
above (Section IV A):
3
volume cell in mm1"
volume examined in mm
= (approx.) 78
F4 =
(If one field were counted, the factor
would be 781, for 100 fields it would
be 7.8.)
NANNOPLANKTON COUNTING
For counting nannoplankton using the high
dry power (10X ocular and 43X objective)
and the "nannoplankton counting cell"
(Figure 9) which is 0. 4 mm deep, a minimum
of 20 separate Whipple fields is suggested.
The same general relationships presented
above (Section IV) can be used to obtain a
multiplier or factor (F5) to convert counts
per 20 fields to counts per ml.
To take another example from Figure 4, at
an approximate magnification of 400X and an
interpupillary setting of 70 (see also Figure 3)
we observe that one side of the Whipple field
measures 0. 260 mm. The volume of the
fields examined is thus obtained as follows:
V5 = side2 X depth X no. of fields
= 0. 26 X 0.26 X 0.4 X 20= .54mm3
and F =
= (approx. ) 1850
It should be noted that the volume of the
nannoplankton cell, . 1 ml, is of no significance
in this particular calculation.
REFERENCES
1 American Public Health Association, et. al.
Standard Methods for the Examination
of Water, Sewage, and Industrial Wastes.
13th Edition. Am. Public Health Assoc.
New York. 1970.
2 Jackson, H.W. and Williams, L.G.
The Calibration and Use of Certain
Plankton Counting Equipment. Trans.
Am. Mic. Soc. LXXXI(1):96-103. 1962.
3 Ingram, W. M. and Palmer, C. M
Simplified Procedures for Collecting,
Examining, and Recording Plankton in
Water. Jour. Am. Water Works.
Assoc. 44(7): 617-724. 1952.
4 Palmer, C. M Algae in Water Supplies.
U. S. D. H. E. W. Public Health Service
Pub. No. 657. 1959.
5 Palmer, C. M. and Maloney, T E. A
New Counting Slide for Nannoplankton.
American Soc. Limnol. and Oceanog.
Special Publications No. 21. pp. 1-6.
1954.
Ill 3-11
-------
Calibration and Use of Plankton Counting Equipment
Area
Uncounted
_ Blrtpt.
Counted
Figure 8
Sedgewick-Rafter counting cell showing bottom scored across for ease in counting
strips. The "strips" as shown in the illustration simply represent the area counted,
and are not marked on the slide. The conventional dimensions are 50 X 20 X 1 mm, but
these should be checked for accurate work.
o
Figure 9
Nannoplankton cell. Dimensions of the circular part of the cell are 17. 9 mm diameter
X 0. 4 mm depth. When covered with a coverglass, the volume contained is 0.1 ml.
The channels for the introduction of sample and the release of air are 2 mm wide and
approximately 5 mm long. This slide is designed to be used with the 4 mm or 43X
(high dry) objective.
6 Welch, Paul S. Limnological Methods.
Blakiston Company. Phila. Toronto.
1948.
7 Whipple, G. C., Fair, G. M., and
Whipple, M. C. The Microscopy of
Drinking Water. John Wiley and Sons.
New York. 1948.
This outline was prepared by H. W. Jackson,
Chief Biologist, National Training Center,
FWQA, Cincinnati, OH 45226.
m 3-12
-------
PREPARATION OF PERMANENT DIATOM MOUNTS
I The identification of many diatoms to
genus and all diatoms to species requires
that the cells be free of organic contents.
This is necessary because the taxonomy of
the diatoms is based on the structure of the
frustule (shells) of the organisms and many
features are masked by the presence of
organic materials which may remain inside.
It is also necessary that at least 1000X
magnification (oil immersion) be used to
detect the structural features used in
identification. No simple procedure for the
accurate routine counting of diatoms has yet
been developed.
II MATERIALS NECESSARY
A Sample Concentration
1 Centrifuge (such as Universal DU)
2 100 ml centrifuge tubes
3 Membrane filter apparatus
4 Vacuum
B Slide Preparation
1 Slides, 1X3 inch, frosted-end
2 Cover glasses, circular #1, 18mm,
0. 13 - . 16 mm thick
3 Resinous mounting medium (such as
Harleco microscope mounting medium)
4 Hot plates
a 1800 F
b 7000 F
5 Disposable pipettes
6 3X6X1/4 inch steel plate
UI PROCEDURE
A The volume of sample needed will vary
according to the density of diatoms and
silt, and only with experience can the
correct sample size be determined. In
most cases, 100 ml will be sufficient.
1 Spin 100 ml at 1000 G for 20 minutes.
2 Withdraw the supernatant liquid with
an aspirator, being careful not to
disturb the concentrate at the bottom
of the centrifuge tube. (Draw off all
but 2-3 ml.)
3 Transfer the concentrate to a labelled
10 ml disposable vial. Label the vial
with a magic marker, diamond pencil,
or "time" label.
4 If the sample has been preserved with
formalin, or contains more than
1.0 gram per liter dissolved solids,
it will be necessary to wash the
concentrate with distilled water. In
this case, transfer the entire concen-
trate to a 15 ml centrifuge tube.
Dilute to 15 ml with distiDed water,
making certain that the sample is well
mixed. Spin for 10 minutes at full
speed in a clinical centrifuge. With-
draw the supernatant liquid, and refill
with distilled water. Spin again for 10
minutes. Withdraw the supernatant
liquid as before, return the concentrate
to the rinsed vial in 2-3 ml of distilled
water and proceed with the mounting.
5 If more than 200 ml of sample must be
centrifuged to obtain sufficient material
to prepare a diatom slide, concentrate
the diatoms by filtering the sample
through a 1. 2 micron pore diameter
membrane filter. Transfer the filter
to a 15 ml centrifuge tube, and dissolve
with 90% acetone. Centrifuge 10
minutes (full speed) and decant with an
aspirator. Refill with 90% acetone.
BI.MIC.enu.lab.5b.6.l
III 4-1
-------
Preparation of Permanent Diatom Mounts
agitate, and spin again for 10 minutes.
Repeat until three fresh acetone washes
have been used. Replace the acetone
with 2-3 ml of distilled water and
transfer to a labelled vial as described
in #4.'
B If the loss of minute forms in supernatant
is suspected, spin 100 ml at 1000 gs in
a batch centrifuge for as long as may be
necessary, then proceed as below.
C Mounting
1 Heat the hot plates to the prescribed
temperatures.
2 Place one cover glass on the steel plate
for each sample.
3 Place the steel plate on the 180° F hot
plate.
4 Transfer a drop of sample to a cover
glass.
5 Allow the water to evaporate (caution:
do not allow it to boil.)
6 Continue to add more sample until a
thin layer of material is noticeable on
the dry cover glass, or until all of the
concentrate has been used. This step
is especially critical, and can be
learned only by trial and error.
7 Transfer the steel plate to the 700° F
hot plate for 20-30 minutes. (The
plate should be hot enough to incinerate
paper.)
8 While the material is on the high
temperature hot plate, label the
microscope slides (use a #2 pencil
or a fine point drawing pen); place
them on the low temperature hot plate,
which now has been reset to approxi-
mately 2750 F.
9 Place a drop of mounting resin on the
microscope slides and allow the solvent
to evaporate.
10 When the incineration of the material
on the cover glasses is complete,
transfer the cover glasses, while still
hot, to the mounting medium.
11 Allow the resin to penetrate the
frustules (1-2 minutes).
12 Remove the slide, place it on a cool
desk top, and press the cover glass
lightly with a pencil eraser for a few
seconds. The medium will harden in
5-10 seconds.
13 Scrape off the excess resin with a
razor blade.
D The preparation is now ready for exam-
ination under an oil immersion objective.
ACKNOWLEDGEMENT:
Certain portions of this outline contains
training material from a prior outline by
M.E. Bender.
This outline was prepared by Dr. C.I.
Weber, Chief, Biological Methods Section,
Analytical Quality Control Laboratory,
1014 Broadway, Cincinnati, OH 45202.
Ill 4-2
-------
DETERMINATION OF ODORS
I INTRODUCTION
Odor shall be determined substantially as
prescribed by the llth edition of "Standard
Methods for the Examination of Water and
Sewage", subject to certain stipulations and
modifications made necessary by the Inter-
national Joint Commission.
The procedure and technique to be followed
are described below.
II REAGENTS AND APPARATUS
A Odor-free water1 - prepared by passing
tap water through activated carbon at a
slow rate of speed. Activated carbon
can be placed at the bottom of a 20-liter
glass bottle. The bottle can be connected
to the tap by rubber tubing leading to glass
tubing above the water. The outlet from
the bottom of the bottle should be glass
tubing. A trap made of inch glass tubing
filled with activated carbon is placed at
the end of the outlet.
500 ml glass-stoppered Erlenmeyer
flasks, each flask with a number. Glass-
ware must be thoroughly cleaned and
rinsed several times with odor-free
water before each use.
C Chemical Thermometer (0-100°C)
10 ml Mohr pipettes, 25 ml graduated
cylinders, 50 ml graduated cylinders,
200 ml graduated cylinders, 500 ml
graduated cylinders. Other pipettes
and cylinders as needed.
One liter glass-stoppered bottles to hold
samples of water being examined. Other
glass bottles and flasks as needed.
Ill PRECAUTIONS
Certain conditions are required to
obtain consistent results. Considerable
practice with the test is desirable to
develop consistent sensitivity to the
sense of smell.
1 In view of the perishability of the
odor test, these determinations
should be made immediately after
collection.
2 The prepared odor-free water should
be truly free of all detectable odor.
3 All glassware must be free of odor.
This is accomplished by thorough
cleansing followed by several rinses
with odor-free water.
4 All dilutions should be compared with
an odorless standard. This aids the
observer in deciding whether air odor
is present or not.
5 All dilutions when examined for odor
should be of a uniform temperature,
deviation not to exceed 1°C.
6 A sudden change in the character of
the odor during the testing procedure
should be considered as a warning
that there may be interference from
outside odors or that the diluting
water may not be odor-free. The
character of odor should always be
recorded for future consideration.
To eliminate psychological influences,
the samples should be coded and in-
termixed so as not to suggest to the
observer what odor concentration is
being observed.
1 Bottles should be colored or covered
with odor-free material or the observer
blindfolded to eliminate auto suggestion
WS. TO. lab. la. 1.66
III 5-1
-------
Determination of Odors
since many samples may possess
color or turbidity.
Test should be conducted in a room
free of outside odors. The observer
should be cautioned to refrain from
smoking or eating for an appreciable
time before taking test. Odors should
be washed from the hands prior to
taking test.
The test should not be prolonged to a
point where the sense of smell becomes
fatigued.
IV PROCEDURE
A To obtain the approximate range of odor
value take 50 ml, 14 ml and 5 ml of sample
and make each sample up to 200 ml with
odor-free water. Compare the odor of
these three with 200 ml of odor-free
water.
1 Cold odor: Bring dilutions to temper-
ature of 24 - 25°C.
2 Shake each flask uniformly before
smelling for odor. Observer should
characterize type of odor.
3 Note which flasks contain odor and
which do not. According to results
obtained, prepare intermediate di-
lutions, in each case using sufficient
odor-free water to make a total
volume of 200 ml.
Include a flask with 200 ml of odor-
free water with each series, as a
blank for comparison.
B Arrange flasks so that their identity is
unknown and bring to desired temperature.
1 Observe for odor and make chart with
a "plus" or "zero" for each dilution.
2 The results are reported in "threshold
odor numbers". The threshold odor
number is calculated from the amount
of sample in the most diluted portion
which gives perceptible odor. The
volume of the dilution (200 ml) divided by
the volume of the sample in the dilution
equals the threshold odor number. For
example, if 5 ml diluted to 200 ml is
the most dilute portion giving perceptible
odor:
200
40, the threshold odor is
numbered 40.
The threshold odor number shall not be
confused with the "threshold odor con-
centration". The threshold odor concentra-
tion is the smallest amount of odor-producing
material in mg/1 required to give perceptible
odor. If the threshold odor concentration is
known, that value multiplied by the threshold
odor number will give the concentration of
the odor-producing material in the sample.
This outline was prepared by E. L.
Robinson, Research Aquatic Biologist,
Fish Toxicology Laboratory, 3411 Church
Street, Newtown, OH 45244.
Ill 5-2
-------
Determination of Odors
ROBERT A. TAFT SANITARY ENGINEERING CENTER
AQUATIC BIOLOGY
ALGAL THRESHOLD ODOR EXPERIMENT
Amount of Culture
Age of Culture
No. Cells per ml_
Mixed. Unialgal, Pure
ml Exp. No.
days Temp. Tested at_
Culture Medium_
Date
Recorde r
Observer No.
Observer
Flask
No.
Culture
No.
Dilution
No.
Threshold Odor No.
Description of Odor
1
R
2
R
3
R
4
R
5
R
6
R
7
R
i
+ = Odor Detected
Remarks
Estimated Composite* T. O. No.
O = No Odor Detected
*Geometric average of T.O. No. of individual observers E.L. R. 1956
III 5-3
-------
COLLECTION AND INTERPRETATION OF BIOLOGICAL LAKE DATA
I INTRODUCTION
A The innumerable interrelationships
of an organism with its environment
are most complicated within the
environs of a lake. The impact of
these interrelationships upon a
lake sampling program and upon the
data collected must be recognized.
B Lake Sampling and Data Collection
Entails:
1 A definition of the problem.
2 A determination of the types of
samples necessary to point to
a solution.
3 A delineation of sampling sites.
4 A judgment on the number of
samples necessary.
5 A decision on the proper time
and periodicity of sample collec-
tions.
6 Some knowledge or understanding of
the science of limnology.
C In any lake sampling program the
primary interest is in the relation-
ship of the aquatic organism with
its environment, and to fully
appreciate this, one must have as
a prerequisite some knowledge of
the science of limnology.
II LIMNOLOGY - The study of inland waters
A Definitions of common terms:
1 Epihmnion - the turbulent super-
ficial layer of a lake lying above
the thermocline which does not
have a permanent thermal strati-
fication.
2. Thermoclme - the layer of water
in a lake between the epilimnion
and hypolimnion in which the
temperature exhibits the greatest
difference in a vertical direction.
3 Hypolimnion - the deep layer of a
lake lying below the thermocline
and removed from surface in-
fluence.
4 Littoral - shorward region of a
body of water; waters edge to the
lake ward limit of rooted aquatic
vegetation.
5 Sublittoral - from lakeward limit
of rooted aquatic vegetation to
the level of the upper limit of the
hypolimnion.
6 Profundal - all of the lake floor
bounding the hypolimnion below
the light controlled limit of
plant growth.
7 Limnetic or Pelagic - region of
free water in which green plants
are present only as phytoplankton.
8 Benthic - the region of the shore
and the bottom of waters.
9 Trophogenic layer - the superficial
layer of a lake in which organic
production from mineral sub-
stances takes place on the basis
of light energy.
10 Tropholytic layer - the deep layer
of the lake where organic dissimilation
predominates because of light de-
ficiency.
11 Oligotrophic - waters with a small
supply of nutrients and, hence, a
small organic production.
W. RE. Ik. 3. 10.66
III 6-1
-------
Collection and Interpretation of Biological Lake Data
12 Eutrophic - waters with a good
supply of nutrients and, hence, a
rich organic production.
13 Dystrophic - bog-type lakes con-
taining yellow-brown water and
much humus.
14, Liebig's law of the minimum -
the yield of a plant or animal
is determined by the quantity
of that particular necessary
substance which is present in
minimal amounts as determined
by the demands of the organism.
15. Seiches - oscillations of the
water level--standing waves.
16. Standing crop - the total amount
of biological growth present in
the water on a selected date.
IH FACTORS WHICH INFLUENCE SAMPLING
AND DATA COLLECTION
A Physical Features
! Temperature
a Lakes are warmed in spring
principally by the action of
wind forcing the warmer water
down into the cooler water
against the forces of gravity.
b Thermal stratification
2. Turbidity
3 Color
4. Water movement
5 Light penetration
a> A factor of turbidity, color,
biological activity, and time
of day
(1) Effective length of daylight
dimemshes with the depth
of the lake.
6 Wind velocity
7. Bottom materials
8. Size, shape, and slope of lake
basin
B, Chemical Factors
1 Alkalinity, pH, and dissolved
minerals excluding nitrogen and
phosphorus
2 Dissolved oxygen ?-
a From photosynthesis in sun-
light
b From contact of lake surface
with the air
c Fluctuates seasonally because
of temperature and biological
activity, and diurnally because
of biological activity.
3 Nutrients for biological growth -
especially nitrogen and phosphorus
a A given body of water will
produce a given quantity of
aquatic life. Biological
production is determined
primarily by the nutrients in
solution in the water, and an
increase in basic fertility will
increase biological activity.
b Basic suppliers of nutrients
include tributary streams,
precipitation from the atmos-
phere, and interchange with
lake bottom sediments.
C Biological Communities
1 The littoral community composed
of rooted vegetation, attached
algae, fish, and a host of inver-
tebrates
2 The limnetic community composed
of fish and plankton
III 6-2
-------
Collection and Interpretation of Biological Lake Data
3 The benthic community composed
of midge larvae, sludgeworms,
fingernail clams, and other bottom
dwelling organisms
IV THEORY AND PRACTICE OF LAKE
SAMPLING
A Purpose of Program
1 To point toward a logical and
satisfactory solution to the
specific problem.
2 To correlate the physical,
chemical, and bib logical pheno-
mena.
3 To reach an understanding of the
interrelationships of the biota
with the environment.
B Types of samples
1 Chemical
a Grab
b Composite
c Other
2. Biological
a Plankton
b, Benthic organisms
c. Fish
d Littoral vegetation
e, Bottom ooze
f Core samples of lake bottom
C Possible sample locations
1 Lake inlets and outlets
2 Random vertical samples in
limnetic region away from littoral
influence.
3 Samples collected at intervals
on line transecting lake basin.
4. Individual "grab"or composited
samples from similar regions
and /or depths of lake.
5 Selected sites to define a specific
problem.
D Number and periodicity of collections
1 The use of statistics is a valuable
and necessary tool both in analyzing
the data and in determining the
number of samples necessary.
2 The question, "How many samples
must I take?", is one of the more
difficult ones facing the investiga-
tor. In reality, the availability
of funds, personnel, and time are
often the determining factors in a
sampling program.
REFERENCES:
1 Birge, E.A., and Juday, C., The Inland
Lakes of Wisconsin. The Plankton. I.,
Its Quantity and Chemical Composition.
Wis. Geol. Nat. Hist. Sur. Bulletin 64,
Science Series 1. No. 13, 222 pp. 1922.
2 Coker, Robert E., Streams, Lakes,
Ponds. The University of North Carolina
Press, Chapel Hill, 327 pp. 1954.
3 Hutchmson, G. E., A Treatise on Limnology.
John Wiley and Sons, Inc., 1,015 pp.
1957.
4 Mackenthun, K. M., Ingram, W. M., and
Ralph Forges. Limnological Aspects of
Recreational Lakes, DHEW, PHS Publi-
cation No. 1167, 1964.
5 Needham, J. A., and Doyd, J. T., The
Life of Inland Waters. Comstock
Publishing Company, Ithaca, New York,
1937.
6 Reid, George K. Ecology of Inland Waters
and Estuaries. Remhold Publishing
Corporation, New York, 375 pp. 1961.
Ill 6-3
-------
Collection and Interpretation of Biological Lake Data
Ruttner, F., Fundamentals of Limnology.
University of Toronto Press., 242 pp.
1953.
Standard Methods for the Examination
of Water and Wastewater. American
Public Health Association, llth Ed.,
626 pp., 1960.
10 Welch, P. S., Limnology. McGraw Hill
Book Company, Inc., 471 pp., 1935.
11 Welch, P. S., Limnological Methods.
The Blakiston Company, Philadelphia,
381pp., 1948.
Symons, J. M., Weibel. S.R., and Robeck,
G.G. Influence of Impoundment on Water
Quality. DHEW, PHS Publication No.
9999 - WP - 18. 1964.
This outline was prepared by K. M.
Mackenthun, Biologist, formerly with
Technical Advisory and Investigations
Activities, FWPCA, SEC.
Ill 6-4
-------
DETERMINATION OF PLANKTON PRODUCTIVITY
I INTRODUCTION
Primary production is the synthesis of organic
matter from inorganic raw materials. The
energy required for this process may come
from light (photosynthesis), or from chemical
sources (chemosynthesis). The primary
synthesis of organic matter in lakes and
streams is carried on by planktonic and ben-
thic algae and bacteria, and aquatic
macrophytes.
II PHOTOSYNTHESIS
The photosynthetic process involves the up-
take of CO2 and the release of Og. The
reactions are enzyme catalyzed and are af-
fected by the following factors:
A Temperature
B Light Intensity
C Light Quality
D pH
E Nutrients
F Trace Elements
III MEASURING PRODUCTIVITY
Methods employed to measure plankton pro-
ductivity are:
A Standing Crop
B Oxygen
C pH
D Carbon-14
IV STANDING CROP METHOD
The productivity of a body of water is indicated,
in a general way, by the density of the plankton
population. The standing crop of plankton is
commonly measured by determining one or
more of the following:
A Dry and Ash-free Weight of Seston
B Cell or Unit Counts
C Cell Volume
D Chlorophyll
E Particulate and Dissolved Carbohydrate
F Particulate and Dissolved Organic Carbon
Increases in the standing crop over a period
of time may be used to determine productivity.
However, this method provides only a rough
approximation of the rate of primary
production.
V OXYGEN METHOD
The use of dissolved oxygen to determine
short-term rates of primary production was
introduced by Gaardner and Gran (1927).
Estimates of the amount of carbon fixed are
based on the premise that one molecule of
oxygen is given off for each atom of carbon
assimilated.
C0
- CH20
A "Light" and "dark" bottles are filled with
sample and resuspended at various depths
for 4-24 hours.
B The concentration of dissolved oxygen is
determined (using the Winkler Method) at
BI. ECO. pro. la. 4.70
HI 8-1
-------
Determination of Plankton Productivity
the beginning and end of the incubation
period. The values obtained are as
follows:
1 Final "light" bottle O2 - initial O2 =
net photosynthesis
2 Initial On - Final "dark" bottle O0 =
respiration
3 Net photosynthesis + respiration =
gross photosynthesis
This method has some serious disadvantages:
A The bottles provide an artificial substrate
for the proliferation of bacteria which
use up large amounts of 0%, resulting in
erroneously high respiration and low net
photosynthesis values.
B The lower limit of sensitivity of the Winkler
Method is 0. 02 mg O% /liter. This is a
serious handicap when working in oligo-
tropic lakes and the open sea.
VI CARBON-14 METHOD
The use of carbon-14 for the measurement
of the rate of carbon assimilation by phyto-
plankton was pioneered by £. Steemann
Nielsen (1952). The method is simple and
very sensitive.
A Carbon-14 labelled sodium bicarbonate
(4 - lOuc /liter) is added to "light" and
"dark" bottles, which are resuspended in
the water for 4-24 hours.
B An aliquot of the sample is passed through
a membrane filter (1.2 jipore diameter),
and the filters are treated with acid to
remove any inorganic labelled carbon.
C The (beta) activity of the filter is deter-
mined with an end-window Geiger tube,
or with gas flow or liquid scintillation
techniques.
D The carbon fixed is determined as
follows:
carbon activity on filter
fixed " total activity added
There are several important disadvantages
in this method.
A Some of the labelled photosynthesis pro-
ducts will be broken down immediately by
respiration, and the liberated carbon-14
reused in photosynthesis. Therefore, it
is generally agreed that the method mea-
sures only net photosynthesis.
B It has been found that the algae rapidly ex-
crete up to 50% of the photosynthate in the
form of organic acids, carbohydrates,
and amino acids. Since these labelled
materials are not retained by the filter,
they escape detection.
VII pH METHOD
The uptake of CO2 by the algae during photo-
syntheses results in an increase in the pH of
the surrounding medium. Periodic pH measure-
ments are made of the body of water being
studied, and the carbon uptake is determined
using published nomographs.
Verduin (1952) used this method in a study
of the productivity of Lake Erie. However,
the method has not gained wide acceptance
because it can be used only in waters with
low alkalinity.
REFERENCES
1 Allen, M. B. Excretion of Organic Com-
pounds by Chlamydomonas. Arch. f.
Mikrobiol. 24-163-168. 1956.
2 Curl, H. Jr., and Small, L. F. Variations
in Photosynthetic Assimilation in Natural
Marine Photoplankton Communities.
Limnol. Oceanogr. 10(Suppl.):R67-R73.
1965.
3 Gaardner, T., and Gran, H. H. Investi-
gations of the Production of the Plankton
in the Oslo Fjord. Rapp. et Proc. -
Verb., Con. Internal. Explor. Mer.
42:1-48. 1927.
X
available
HCO;
X
correction for
isotope
discrimination
HI 8-2
-------
Determination of Plankton Productivity
4 Goldman, C. R. Molybdenum as a Factor
Limiting Primary Productivity in
Castle Lake, California. Science 132:
1016-1017. 1960.
5 Kamen, M. D. Primary Processes in
Photosynthesis. Academic Press,
New York. 1963.
6 Marshall, S. M., and Orr. A. P. Carbo-
hydrate as a Measure of Photoplankton.
J. Mar. Biol. Assoc. U.K. 42:511-519.
1962.
7 Ryther, J. H. Photosynthesis in the Ocean
as a Function of Light Intensity.
Limnol. Oceanogr. 1:61-70. 1956.
8 Steemann Nielsen, E. The Use of Radio-
active Carbon (C-14) for Measuring
Organic Production in the Sea. J. Con.
Internal. Explor. Mer. 18:117-140. 1952.
9 Strickland, J. D. H. Measuring the
Production of Marine Phytoplankton.
BuU. Fish. Res. Bd. Can. No. 122:
1-172. 1960.
10 Verdum. J. Photosynthesis and Growth
Rates of Two Diatom Communities in
Western Lake Erie. Ecology 33(2):
163-168. 1952.
11 Vernon, L. P. Bacterial Phytosynthesis.
Ann. Rev. Plant. Physiol. 15:73-100.1962
12 Wetzel, R. G. A Comparative Study of the
Primary Productivity of Higher Aquatic
Plants, Periphyton, and Phytoplankton
in a Large, Shallow Lake. Intern at.
Rev. Hydrobiol. 49:1-61. 1964.
13 Yentsch, Charles S. The Measurement
of Cnloroplastic Pigments- Thirty
Years of Progress? pp. 255-270 in
Chemical Environment in the Aquatic
Habitat. Proc. IBP Symposium.
Amsterdam. 1967. (N.V. Noord-
Hollandsche Uitgevers Maatschappij.
Amsterdam, Netherlands. 8.95)
This outline was prepared by C. I. Weber,
Chief, Biological Methods Branch,
Analytical Quality Control Laboratory,
1014 Broadway, Cincinnati, OH 45202.
HI 8-3
-------
AERIAL RECONNAISSANCE IN POLLUTION SURVEILLANCE
I INTRODUCTION
A Definition
The word "reconnaissance" is derived from
the word "reconnoiter" which means to
conduct a preliminary examination or survey.
Its earlier applications to engineering and
military requirements has been expanded
to include photomapping and interpretations
of natural resources. Aerial reconnaissance
can be defined as "airborne examination or
survey procedures performed by heavier-
than-air craft, lighter-than-air craft, or
earth orbiting satellites. "
B Types of Aerial Reconnaissance
1 Visual
Examination of the flight path by a human
observer with no provision for permanent
recording for later study. This form
of aerial observation has limited use but
in many cases can complement the other
forms.
2 Image forming sensor
Image recording of the covered flight
path where maximum advantage can be
made of image interpretation techniques.
Image forming sensors includes such
instruments as cameras, infrared
scanners, and radar.
3 Nommage forming sensors
Nommage forming sensors include such
devices as oscilloscopes, strip charts,
and dial indicators which directly indicate
parameter differentials as received by
the sensing elements from the target scan.
4 Combination image forming and nonimage
sensors
It is advantageous at times to combine
both of these techniques in order to
rapidly interpret flight path imagery.
This advantage is shown in Figure 1
where a heated effluent discharge is
traced to a canal by infrared imagery
and indicated by the "lighter" plume
which parallels the shoreline. The
oscilloscope phototracing indicates the
temperature differential of the discharge
point as compared to the offshore bay
area.
C Detected images can be identified by one
or more of the following criteria.
1 Size of image
2 Shape of image
3 Tonal qualities of image
4 Image profile shadowing
5 Location of image
6 Texture patterns of grouped images
7 Spatial relationship of image to
surrounding bodies
D Selection of Remote Sensing Method
1 A knowledge of the electromagnetic
spectrum (Figure 2) is important in
order to select the most advantageous
method of remote sensing. It will be
noted that there are transmissibility
and sensitivity limits as well as existing
film limits with respect to equipment
wave length patterns.
2 Remote sensing methods can be
categorized into two areas.
a Source active: Utilization of an
instrument which is capable of
emitting a source of energy which
is transmitted to the target area and
emanations are received whose
characteristics are dependent upon
the nature of the specific target.
WP.SUR.fm.6.8.69
HI 10-1
-------
Aerial Reconnaissance in Pollution Surveillance
An example of such a device is the
Radar set (Figure 3) or special
Infrared Emitters both of which are
capable of being utilized in total
darkness.
b Source passive: Utilization of
instrumentation which is only capable
of receiving target area emanations.
Such remote sensing instruments,
therefore, do not transmit artificial
energy sources but depend upon the
sun as a source of energy which is
selectively received and individually
reflected by each object in the target
area. Aerial photography and most
of the remote sensing is by this
method and the included regions of
the electromagnetic spectrum are the
upper portions of the UV band to the
near infrared region.
Selection of desired wavelength is
dependent upon the optimum emission
capability of the instrument which has
the shortest wavelength possible to
sharply differentiate small objects while
being long enough to preclude excessive
energy "scattering" and yet not long
enough so that photographic recording
is impossible.
II TECHNIQUES
A Radar(RAdio Detection And Ranging)
Radar is particularly useful for discerning
certain types of vegetation which may not
appear in the best visual quality in
Panchromatic Color Film (sensitive to the
entire visible spectrum) and also has all-
weather and around-the-clock capabilities.
Radar in the longer wavelength ranges is
capable of penetrating dense vegetative
cover and this advantage has been found
to be useful in detecting drainage networks
and geological features.
B Conventional Photography
Conventional photography covers the entire
range of the visible spectrum and employs
both black-and-white film and color film.
Some of the special films are capable of
photographing a portion of the infrared band
(the "near" infrared region) and this
capability is useful in special situations.
-------
Aerial Reconnaissance in Pollution Surveillance
ELECTROMAGNETIC SPECTRUM
WAVELENGTH
3m 300km
I I I I I I I I I I I I I I I
AMM/
RAY
1 1
1 1
20
10
k uv
X-RAY
1
1
18
10
1 1
1 1
1
S
1
L
E
IR
1 1
r i i
16 14
10 10
RADAR UHF
(MICROWAVE)
1 1 1 1 1
1 1 1 1 1
12 10 8
10 10 10
RADIO
1
1
1
1
6
10
LF
1 1
1 1
AUDIO
1
1
4
10
AC
1 1
FREQ
1 1
2
10
1
UE
1
i
VISIBLE LIGHT (LIMITS OF HUMAN VISION)
A.
(cps)
THE PHOTOGRAPHIC
WAVE ANGSTROM UNITS
LENGTH
§ § §
°. o o
0. m rx
INFRA
RED
SPECTRUM
(1 Angstrom Unit1
o
0
o
0
III II
| (Red Orange Yellow
'0 1 Millimicrons = 10 ^
§ §
° °.
i 7 i ii
Green BlueVioletl
Centimeters)
o
0
o
1"
ULTRA
VIOLET
§
0
cJ
TO 11,000 A
TO 12,000 A
PHOTOTUBE SENSITIVITY
SPECIAL FILMS
h
ORDINARY FILM-
ORTHO FILM
PANCHROMATIC FILM
I TRANSMISSION LIMIT
hGLASS LENSES
FIGURE 2
-------
Aerial Reconnaissance in Pollution Surveillance
Target
FIGURE 3
1 Direct photomapping and panoramic scan
Figure 4 illustrated the manner in which
direct photomapping differs from the
panoramic scanning technique and it can
be seen that continual attached mapping
scans can be made of all or portions of
the flight path to be analyzed.
2 Multiband photography
Multiband photography is, as its name
implies, the synchronized photography
of up to nine separate photographs of the
same target area each of which is
photographed in a different wavelength.
A distinct advantage of this method is
that, during photo analysis, distinct
anomalies may be evident between
objects appearing in the different bands
and further investigation may be
warranted. An example of this is shown
in Figure 5 where it was observed that
an anomaly in vegetative growth was
apparent in the Infrared wavelength
while not being apparent in the other
visual spectral ranges. Further
investigations proved that gravel washings
in slug intervals had affected normal
growth of the streamside vegetation.
3 Infrared photography
Infrared photography has been found
useful in depicting certain hydrological
features. Within the photographic
ranges it is not a heat-measuring tool
and therefore cannot detect thermal
anomalies within bodies of water but
instead sharply delineates shoreline and
tributary features and has haze
penetrating qualities. The blues appear
much darker by infrared photography
and the reds, greens, and yellows will
show up much lighter when compared to
a panchromatic color photograph.
-------
Aerial Reconnaissance in Pollution Surveillance
Direct
Photomapping
FIGURE 4
impaired or
' dead vegetation
normal
vegetation
Infrared Photograph
FIGURE 5
-------
Aerial Reconnaissance in Pollution Surveillance
4 Gamma Ray Spectrometer
This device functions in the very short
wavelengths and has been found to have
excellent detecting capabilities for
radioactivity. Such a device would serve
as an excellent means of searching for
radioactive waste spills which have
occurred either accidentally or
intentionally.
C Infrared Radiometry
Infrared imagery is different than infrared
photography since the tone of the imagery
is directly related to the infrared radiation
emitted from the target area. This type
of scanner converts the normally
unphotographic IR band to a thermal
imagery. An optical-mechanical device
is necessary since the normal thermal
energy emitted by the camera itself will
influence radiations received from the
target area and this elimination of detector
internal radiation is accomplished by
miniturization and application of extremely
low temperatures. One example of the
usefulness of infrared imagery is shown in
Figure 6 where the dark areas along the
shoreline were found to be caused by cold
water infiltration by a previously unknown
source. Such a finding could be indicative,
for example, of a new untapped freshwater
source or a polluting influence from a
remote source. This cold water infiltration
was not evident by normal visual spectrum
photography and the IR photograph would
only indicate a more distinct shoreline.
D Automatic Analysis
In the analysis of large numbers of prints
use can be made of photoelectric scanners
wherein automatic recording of degree of
brightness can be recorded on a tape
printout. An example of this procedure is
shown in Figure 7 where each symbol has
a significance relating to "tone signature. "
The "D" zones are the darker areas while
the other symbols relate to lighter zones.
In the center the clear area is a river
tributary. Future predictions are that
automatic encoding can be channeled to
computers wherein decisions can be made,
for instance, in water management policies.
E Stereoscopic Analysis
The principles and methods developed
during World War II for the stereoscopic
analysis of aerial photographs has been a
continuing and expanding science. Its
principles are based upon the fact that the
human eyes are normally spaced about
65 millimeters apart and are capable of
compositing, via the optic nerve, the two
separate and distinct images viewed by
each orbit. Aerial stereoscopic analysis
depends upon the placing of two photographs
precise distances apart with precise
overlapping as required by normal human
visual acuity. With the aid of special
stereoscopic viewing lenses the viewer is
able, in most cases, to composite both
of the photographs in the same manner as
with normal vision and thus a sense of
depth can be imparted materially enhancing
the analyzing of the terrain.
Ill APPLICATIONS
Aerial reconnaissance in pollution sur-
veillance can be accomplished in a variety
of ways and the method chosen usually is
dictated by costs involved and what is
desired with regard to immediate or
potential information.
A Visual Observations
The simplest and least expensive method
is to visually observe the study area and
this can be useful in determining sampling
sites, locating previously undetermined
tributaries, pinpointing visual gross
pollutant effluents, etc. To augment
these observations it is usually better
to have a permanent recording of the
survey area in the form of photographic
or imagery recordings.
B Photographic Techniques
Photographic evidence, either color,
black-and-white, or special film, can be
recorded in various attitudes or positions
of the aircraft in order to make best use
of interpretation techniques such as
stereoscopic analysis.
Ill 10-6
-------
Aerial Reconnaissance in Pollution Surveillance
FIGURE 6
)) XXX "tin.ODD D
D D D n D n :
"\v r> ,,, --"-"- r D D X XX D D D D D D
J pppC ff",f CC DD ^^ D '' D D D '
( cw/ C tt*£fe ::- Cp r,D XCC DD DD D D
( ( ) ) "-::- -" CD D D ODD C C DDD DDL
( -::-::- ;:-::- DDDD ^ 'JDD D D D D L
,0
v-
0
DDDD
DO
D
D
J
D DDDD' (
DDDD DDDD (
D D D DDiO ()) )
DD D D D X CC (( )
DDDE X XCCC C ( )
D X XXX X (())((
**»*c c c ** 0) ) ) 0)
** * CC XX -" ( )) ( ) XX ( )
--"- C X "- CC ( ) ) )) )
C XXX "-C ()((())) 0
X » D ( ) ( ^ ( ) )
X D D D D D DDD D D D D D
X D D D D CCDDD D D D D D
, DDtD D DDDL
'CD D D D D D ,
DDDDD OCX DDD D D D DC
D L
D D
(
3D D
DD D D D L
FIGURE 7
-------
Aerial Reconnaissance in Pollution Surveillance
1 Current patterns and flow velocity can
be ascertained by a method developed
by the U.S. Coast and Geodetic Survey
whereby powdered aluminum is surface
distributed to a wide area and subsequent
photographic patterns can be analyzed.
Flowing bodies can either appear as
"depressions" or "elevations" depending
upon direction of flow with relation to
the photographing aircraft and from these
"parallax" anomalies the velocity can
be determined.
2 Photographic IR has found applications
in the areas of detecting sharp
delineations of shorelines and tributaries
especially when haze is prevalent.
3 The work of Strandberg concludes that
"In aerial photographs, oxygen-deficient
water frequently appears black, or at
least darker in tone and this appearance
may be caused by the incomplete reduction
of wastes by anaerobic bacteria. " Thus
his black-and-white photographs of
waterbodies with low values of dissolved
oxygen (DO) following a source of waste
discharge are characterized by a plume
discharge followed downstream by dark
water tones.
Further values ascribed to photography
in this manual include color photographs
dealing with fish kills where the extent
of damage can be better ascessed and
the use of "false" color infrared
photography where algal masses can be
differentiated and, again, the low DO
concentration is manifested by a color
anomaly.
C Imagery Techniques
1 Utilization of the "mid" and "far"
infrared wavelengths in the form of
"imagery" has been found useful in
analyzing for thermal pollution and its
concomitant circulation and diffusion
patterns for which "thermographs" can
be described for large water bodies.2
It is also possible to establish naturally
occurring patterns of seasonal and
diurnal thermal ranges for surface waters.
It has been ascertained, in principle,
that it is possible to identify pollutants
by study of spectral emissivity
characteristics for particular wave-
lengths and this can be of value, for
instance, when one is searching for a
particular pollutant and the emissivity
characteristics for the waterbody can
be compared with known patterns for
the specific pollutant. 2
Utilization of radar and orbiting
satellite platforms can produce imagery
of great usefulness. The radar technique
can precisely delineate shoreline
characteristics without vegetative
interferences and thus can thoroughly
image drainage basins. Satellite
imagery and photography can obtain the
same information but on a much broader
scale and with great rapidity. Recent
lines of thought have postulated that this
orbiting scanner can be fitted with
computing devices to render decisions
upon analysis of its constant data
collection. 3 Such decisions can be of
immeasurable aid to the water con-
servationist who may require long
range predictions based upon analysis
of whole river basins.
IV SUMMARY
Development of aerial reconnaissance in
pollution surveillance has proceeded in the
areas of technique and interpretation. With
regard to the techniques available most of
the earlier problems have been overcome to
a degree where available material is amenable
to analysis. Such early problems as pitch
and yaw of the aircraft, cloud cover, imagery
instrumentation, etc., has been overcome
to a large degree by available engineering
skills and the need for improvement due to
military needs. In the area of interpretation,
however, much has yet to be accomplished
to develop this tool as an extension of
laboratory analysis which it will augment
rather than replace.
Ill 10-8
-------
Aerial Reconnaissance in Pollution Surveillance
REFERENCES
1 Strandberg, C.H. "Aerial Discovery
Manual. " John Wiley &. Sons, Inc.,
New York.
2 Van Lopik, J. R., Ramble, G.S. and
Pressman, A.E. "Pollution Surveillance
by Noncontact Infrared Techniques."
Jour. Water Poll. Control Fed., 40, 3,
425, March 1968.
3 Colwell, R. N. "Remote Sensing of
Natural Resources. " Scientific
American. January 1968.
This outline was prepared by R. Russomanno,
Microbiologist, National Training Center,
FWPCA, Cincinnati, OH 45226.
ffl 10-9
-------
LABORATORY EXERCISES
Proportional Counting of Plankton 11
Calibration of Plankton Counting Equipment 12
Fundamentals of Quantitative Counting 13
Class Problem in Plankton Analysis 14
-------
LABORATORY: PROPORTIONAL COUNTING OF PLANKTON
I OBJECTIVE
To learn and practice the techniques of
proportional counting of mixed plankton
samples.
H MATERIALS
A Several plankton samples, each containing
a number of plankton forms.
B Class slides, cover slips, and dropping
pipets.
HI PROCEDURES
A Make an ordinary wet mount of the
sample provided.
B Scan the slide. Identify and list all types
of plankton present.
C Proportional Counting (use clump count)
1 Field count
a Count and tally all individuals of
each type present in a field. The
best way to do this is to list the
most common types separately and
record the counts and then enumerate
the other forms.
b Move the slide at random and
repeat the process. Do this for
5 or 10 fields.
c Tally the results and compute the
percent of each type.
2 Five hundred count
a Moving the slide at random count
and tally all the types of plankton
as before until a total of 500 cells
or clumps have been counted.
b Tally the results and compute the
percentage of each type as before.
IV RESULTS
A Record your results for both methods
on the board.
B Discuss the two methods and the use of
the proportional count results.
This outline was prepared by M. E. Bender,
Biologist, formerly with FWPCA Training
Activities, SEC.
BI.MIC. enu. lab. 6a. 8.69
in 11-1
-------
LABORATORY: CALIBRATION OF PLANKTON COUNTING EQUIPMENT
I OBJECTIVES
A To Become Familiar with Microscope
Calibration Procedures
B To Calibrate the Particular Equipment
Assigned to you
II MATERIALS
A Whipple, Plankton Counting Reticule
B Compound Microscope as Assigned
C Stage Micrometer
III PROCEDURE
A Adjust the interpupillary distance to the
position most comfortable for your eyes.
and record the setting on the "Microscope
Calibration Data" sheet.
B Install a Whipple plankton counting reticule
in the right eyepiece.
C Obtain a stage micrometer and focus on the
scale at 100X magnification.
D Record the exact dimensions of the entire
field in the column marked "Whole" on the
plate "Microscope Calibration Data. "
E Do the same with the 200X and 400X
magnifications.
F Return the stage micrometer to the supply
table.
G Values for the "Large" and "Small"
columns may now be calculated
arithmetically. There are ten large
squares across the whole field, and 5
small squares across the large square
which is subdivided, in the center of the
field.
H Calculate the conversion factors to counts
per ml according to the formulae in the
lecture entitled "Calibration and Use of
Plankton Counting Equipment. "
This outline was prepared by H. W. Jackson,
Chief Biologist, National Training Center,
FWPCA, Cincinnati, OH 45226.
BI. MET. mic. lab. la. 5. 70
III 12-1
-------
Laboratory: Calibration of Plankton Counting Equipment
MICROSCOPE CALIBRATION DATA
Microscope No.
Approximate
Magnification
Tube
Length, or
Interpupillary
Setting
Linear dimensions of Whipple
squares in millimeters*
Whole
Large
Small
Factor for
Conversion
to count/ ml
100X. obtained with
Objective
Serial No.
and Ocular
Serial No
200X. obtaine
Objective
Serial No.
and Ocular
Serial No.
(2 S-R Strios)
d with (2 s-R Strios)
.-_ . (Nannoplankton)
400X, obtained with (cell- 20 fields )
Objective
Serial No.
and Ocular
Serial No.
*lmm = 1000 microns
BI.AQ.pl 8 10. 60.
Ill 12-2
-------
Laboratory: Calibration of Plankton Counting Equipment
MICROSCOPE CALIBRATION DATA
Microscope No
Approximate
Magnification
Tube
Length, or
Interpupillary
Setting
Linear dimensions of Whipple
squares in millimeters*
Whole
Large
Small
Factor for
Conversion
to count /ml
100X, obtained with <2 S-R Strips)
Objective
Serial No.
and Ocular
Serial No
200X, obtainei
Objective
Serial No.
and Ocular
Serial No.
400X, obtaine
Objective
Serial No.
and Ocular
Serial No.
J with
d with
-
(2 S-R Strips)
(Nannoplankton)
(cell- 20 fields )
*lmm = 1000 microns
BI. AQ. pi 8 10. 60.
Ill 12-3
-------
LABORATORY: FUNDAMENTALS OF QUANTITATIVE COUNTING
I OBJECTIVE
To learn and practice the basic techniques of
quantitative plankton counting
II MATERIALS
A Plankton Samples Containing a Variety of
Plankton Forms
B S-R Cells and Coverglasses, Large Bore
1 ml Pipettes, Whipple Discs, Plankton
Record Form
III PROCEDURE
Fill the S-R cell with sample number 1
follows:
as
Place the coverglass diagonally across
the S-R cell This leaves the other two
corners uncovered; one for putting in
the sample fluid, the other to allow
air to be driven out as it is replaced
by the incoming aliquot. Shake the
sample to disperse the plankton. Before
settling occurs in the sample draw about
1-1/4 ml of the fluid into the pipette
and quickly fill the S-R cell by delivering
the aliquot into one of the open corners
of the chamber.
C Starting from one end of the S-R cell and
preceding to the opposite (this is called a
strip count, begin counting (clump counts)
the plankton forms. The length of the
cell may be traversed in several ways.
1 Count all the forms in the Whipple
square or in a portion of the square,
record the count and move the slide so
that the square covers the adjoining
area.
2 Move the slide very slowly counting
and recording the various forms as
they pass the leading edge of the
Whipple disc.
IV RESULTS
A Using the correction factor obtained in
the previous laboratory compute the
number of plankton organisms per ml.
B Record the results on the board.
C Discussion of Results
D Refill the slide with a fresh aliquot and
recount the sample. Compare results
with the first count.
E Count the other samples of mixed plankton
as assigned, following the same procedure.
Using lOOx focus on the sample. After
focus has been obtained switch to 200x.
Scan the slide and list the plankton forms
present.
This outline was prepared by M.E. Bender,
Former Biologist, FWPCA, Water Pollution
Training Activities, SEC.
BI. MIC. enu. lab. 7.6. 68
III 13-1
-------
Laboratory: Fundamentals of Quantitative Counting
riANKTON COUNT IECOID
Body of witcr
Dau Coltoeud
. DIM Amljrnid.
Dopth_
TOTAL COUNT
Difft rvntlal
Count |
Notes. Talleyi
Total
Vol, Area,
or Typo of Count
FIELD CONDITIONS
(feather toda}
Previous Wont her
Turbidity Moth*
Method of Collo
i T MadlBff
tlan
Total Vol Collaeted
Preservative
Fllaawntoti
Other Plan
Surface 8c
Dead Fl«h
. Alu.
Other "hyilc
al or CIWBlcal Dat
HiRniflcatlon
LABORATORY
Method of Prc
Dopartun frc
Significance
Tret taw nt Rt
Kultl factor
ANALYSIS
1
per
11 tor
Totalai
Boutin* _
puurr on OTHER DATA, ran s
Other Cheatlc
Reejjlt of Jjp
X
tatawnt
Taate and Odor
D._
niter Run*
Oltmr
SUGGESTED DASIC K)U TOR PLANKTOH MCODD3
IIWJ
4/M
ni HIC fnu pi 2 4 68
III 13-2
-------
O .-.
CHAPTER IV
INTERPRETATION AND SIGNIFICANCE OF PLANKTON
Algae and Actinomycetes in Water Supplies
Algae as Indicators of Pollution
Public Health Significance of Toxic Algae
Odor Production by Algae and Other Organisms
Organic Enrichment and Dissolved Oxygen Relationships
in Water
Plankton in Oligotrophic Lakes
The Effects of Pollution on Lakes
1
2
3
4
5
6
7
-------
ALGAE AND ACTINOMYCETES IN WATER SUPPLIES
I Water treatment always should include
detection and control of microorganisms.
A Two types of microorganisms are involved:
1 Pathogenic types include such forms as
the typhoid bacteria, the dysentery
ameba, and the infectious hepatitis
virus.
2 Interference types include taste and
odor organisms, filter-clogging
organisms, pipe-infesting organisms.
and others.
B Water treatment practices are closely
associated with these organisms.
1 For pathogens, practices include
coliform tests, use of chlorine, and
guarding the water supply against fecal
pollution.
2 For interference organisms, practices
include plankton enumeration, use of
copper sulfate and the covering of
reserviors.
3 Many of the other treatment practices
have significant effects on the organisms.
At Indianapolis, copepods were present in
parts of the distribution system in numbers
sufficient to be visible in the drinking
water. The eggs of the copepods were
found to pass through the filters and to
hatch in the distribution system.
At Oklahoma City, prominent earthy odors
have appeared frequently. The organisms
blamed for this trouble are the mold-like
actinomycetes.
At Peoria, white wigglers up to 3/8" long
were reported in the tap water, during
early March, 1956. These chironomid
larvae had hatched in the city's open
reservoir, requiring that the reservoir be
drained, cleaned and treated with a larvicide.
At Chicago, diatoms are a very important
cause of short filter runs. The one diatom
Tabellaria is considered to be more
responsible than any other organism for
this trouble.
In Ontario, the alga Cladophora often
grows in large numbers attached to rocks
on the shoreline of lakes. When the alga
is broken loose it collects near the shore-
line and gives rise to very offensive odors.
C This discussion will be limited to the inter-
ference organisms.
II EXAMPLES OF PROBLEMS CAUSED BY
INTERFERENCE ORGANISMS
A At Chicago, the alga Dinobryon reappears
almost every year, generally in June and
July in numbers sufficient to impart a
prominent fishy odor to the water. In
1951, it required an estimated $70, 500
worth of activated carbon to control the odor
of this organism for a period of two months.
G In a water supply impoundment in Utah the
plankton algae frequently cause the pH of
the water to increase to 8. 3 or higher,
requiring that the water be treated with
acid to obtain the desired pH of 8 or lower.
H In Texas a water supply from underground
sources was stored in a large open settling
basin. Oscillatona and unicellular green
algae developed in large numbers in the
stored water, turning it green and pro-
ducing a strong odor.
I Los Angeles has more than 25 open reser-
voirs of various sizes and ranging in
elevation from almost sea level to over
BI. MIC. 12c.3.70
IV 1-1
-------
Algae and Actinomycetot in Water Supplies
III
7,000 feet. Many tons of copper sulfate
are used every year in these reservoirs
for rigid control of plankton, chiefly
diatoms and occassionally blue-green
algae. This treatment is carried out to
improve the water quality including the
reduction of tastes and odors.
TYPES OF PROBLEMS CAUSED BY
INTERFERENCE ORGANISMS
A Tastes and Odors
1 May be caused by algae, actinomycetes,
Crustacea, and anaerobic bacteria.
2 Common algal odors imparted to water
are ones described as fish, earthy,
musty, grassy, cucumber, geranium,
nasturtium, and septic.
3 Common actmomycete odor is earthy.
4 Tastes produced in water by algae
include sweet and bitter.
5 Other causative agents of tastes and
odors may be industrial wastes, sludge,
and compounds dissolved from soil and
rock, and chemicals used in treatment.
B Filter Clogging
1 Both rapid and slow sand filters are
affected.
2 Diatoms are the organisms most
frequently involved but blue-green
algae, filamentous green algae and
other organisms as well as silt may
cause it.
C Other Problems in the Treatment Plant
1 Algae may cause variation in the pH,
hardness, color, and organic content
of the water.
2 Amount of plankton organisms often
influences the rate and effectiveness
of coagulation.
3 Chlorine dosage may depend upon
amount of plankton organisms present.
4 Growths of algae may reduce the flow
through influent channels and screens.
5 Organisms may be responsible for
increasing the quantity of sludge to be
disposed of in sedimentation basins.
6 Microcrustacea "spot" paper in paper
mill rolls.
D Infestation of Distribution Systems
1 Attached organisms reduce the rate of
flow in the pipes.
2 Iron and sulfur bacteria may initiate
or stimulate corrosion of pipes.
3 Organisms may appear as visible
bodies in tap water.
4 Tastes and odors may result from
presence of organisms.
5 Chlorine residual is difficult to main-
tain when organic matter is present.
6 Organisms could theoretically harbor
and protect against chlorine certain
pathogenic bacteria.
E Profuse Growths of Organisms in Raw
Water Supplies
1 A limited and balanced growth of
various organisms is generally an
asset.
2 Extensive surface mats, blooms and
marginal growths often cause troubles
along the shoreline and eventually in
the treatment plant.
3 Some fish kills may be caused by
profuse growths of algae by reducing the
DO during the night.
4 Certain massive growths of blue-green
algae are deadly poisonous to animals.
IV ORGANISMS INVOLVED
A Animal forms include protozoa, rotifers,
crustaceans, worms, bryozoans, fresh water
IV 1-2
-------
Algae and Actinomycetes in Water Supplies
sponges, water mites and larval stages of
various insects.
Plant forms include algae, actinomycetes
and other bacteria, molds and larger
aquatic green plants.
V IMPORTANCE OF BIOLOGICAL
PROBLEMS
A The increased use of surface water supplies
increases the problems caused by organ-
isms. Biological problems are less
common with ground water supplies.
B Standards of water quality requested by
domestic and industrial patrons are rising.
C Procedures for detection, control and
prevention of problems caused by organisms
are improving and are receiving more
extensive use.
VI A number of methods may be used to
control the interference organisms or their
products:
A Addition to water or an algicide or pesticide
such as copper sulfate, chlorine dioxide
or copper-chlorine-ammonia.
B Mechanical cleaning of distribution lines,
settling basins, sand filters, screens, and
reservoir walls.
C Modification of coagulation, filtration,
chemical treatment, or location of intake.
D Use of absorbent, such as activated
carbon, for taste and odor substances.
E Modification of Reservoir to Reduce the
Opportunities for Massive Growths of
Algae
1 By covering treated water reservoir to
exclude sunlight
2 By increasing the depth of the water in
reservoirs
3 By eliminating shallow marginal areas
4 By reducing the amount of fertilizing
nutrients entering the reservoir.
VII It is generally more satisfactory to
anticipate and prevent problems due to these
organisms than it is to cope with them later.
A Routine biological tests are essential to
detect the initial development or presence
of interference organisms.
1 Control measures can then be used
before problem becomes acute.
2 These tests should be applied to the
raw treatment plant water supply and
distribution system.
B In the Reservoir or Other Raw Water Supply
1 Routine plankton counts should be made
of water samples from selected loca-
tions. Plankton counter should be
aware of the particular organisms
known to be most troublesome.
2 During the warmer months routine
surveys of the reservoir, lake or
stream should be made to record any
visible growths of algae and other
organisms.
3 Odor tests of water from several
locations should be made to obtain
advance notice of potential trouble at
the treatment plant.
C In the Treatment Plant
1 Records of plankton counts and threshold
odor between each step in treatment
gives data on effectiveness of each
procedure.
2 Coagulation and filtration can be
adjusted to remove up to 95% or more
of organisms in water.
IV 1-3
-------
Algae and Actinomycetes in Water Supplies
3 Microscopic analysis of samples of
filter material for organisms can
supply data useful in modifying sand
filtration and treatment of finished
water.
D In the Distribution System With Its
Finished Water
1 Open reservoirs require constant
attention especially during summer.
2 Parts of the system farthest from the
treatment plant or adjacent to dead
ends require most frequent sampling
for organisms and tastes and odors.
VIII SUMMARY
A Interference organisms cause problems
in distribution systems, treatment plants,
raw water supplies.
B Organisms involved include algae, actino-
mycetes, other bacteria, and minute
aquatic animals.
C Control is by special chemicals, mechanical
cleaning, adjustment of chemical or
mechanical treatment and by modification
of reservoirs, intakes, etc., for the raw
water supply.
D Facilities for detection of problems in
their early stages are required for most
efficient and satisfactory control.
REFERENCES
1 Palmer, C. M. Algae in Water Supplies.
An Illustrated Manual on the Identification,
Significance, and Control of Algae in
Water Supplies. U. S. Public Health
Service Publication No. 657. 1959.
p. 88.
2 Palmer, C.M. and Poston, H.W.
Algae and Other Interference
Organisms in Indiana Water Supplies.
Jour. Amer. Water Works Assn.
48:1335-1346. 1956.
3 Palmer, C.M. Algae and Other Inter-
ference Organisms in New England
Water Supplies. Jour. New England
Water WorksAssn. 72:27-46. 1958.
4 Palmer, C.M. Algae and Other Orga-
nisms in Waters of the Chesapeake
Area. Jour. Amer. Water Works
Assn. 50:938-950. 1958.
5 Palmer, C.M. Algae and Other Inter-
ference Organisms in the Waters of
the South Central United States. Jour.
Amer. Water Works Assn. 52:897-
914. 1960.
6 Silvey, J.K. and Roach. A.W.
Actinomycetes May Cause Tastes
and Odors in Water Supplies. Public
Works 87. 5:103-106,210,212. 1956.
7 Ingram, W.M. and Bartsch, A.F.
Operators Identification Guide to
Animals Associated with Potable
Water Supplies. Jour. Amer. Water
WorksAssn. 52:1521-1550. 1960.
8 Otto, N. E. and Hartley. T.R. Aquatic
Pests on Irrigation Systems.
Identification Guide. Bur. of
Reclamation. USDI. 72 pp. 1965.
9 Herbst, Richard P. Ecological Factors
and the Distribution of Cladophera
glomerata in the Great Lakes.
Amer. Midi. Nat. 82:90-98. 1969.
This outline was prepared by C.M. Palmer,
formerly Aquatic Biologist, In Charge,
Interference Organism Studies, Microbiology
Activities, Research & Development,
Cincinnati Water Research Laboratory,
FWPCA.
IV 1-4
-------
ALGAE IMPORTANT IN WATER SUPPLIES
TASTE AND ODOR ALGAE
.^.r
-*v- -..
; . . , .
£/:: .
1- *::'
|;-Y.;:;.-
V.--'1:-'.-.
V:;:.::-
^
^ *
%
t
* .
»
x
A
* . * ^
::"x*
"~ *i
. r
.«
;
PLATE I
-------
FILTER CLOGGING ALGAE
CHROOCOCCUS
PLATE 2
-------
POLLUTED WATER ALGAE
PLATE 3
-------
CLEAN WATER ALGAE
CLAOOPHOBA
PLATE 4
-------
SURFACE WATER ALGAE
SCENEOfSMUS
PLATE 5
-------
ALGAE GROWING ON RESERVOIR WALLS
PLATE 6
-------
ALGAE AS INDICATORS OF POLLUTION
I LIMITATIONS
A Algae are only one of a number of types
of organisms present which could be
considered.
B Forms recognized here as algae are
comparatively simple, pigmented, aquatic
organisms, including blue-greens, greens,
diatoms and pigmented flagellates.
C Various pollutants react differently on
algae. Organic pollutants such as house-
hold sewage will be dealt with here.
D No algae are intestinal organisms. They
therefore are not indicators of pollution
in the same way that coliform bacteria
are.
B Wastes may have physical effects on
certain algae. May cause plasmolysis,
change in rate of absorption of nutrients,
etc.
C Wastes may reduce available light,
increase the water temperature, and
cover up the areas for attachment to
rocks.
D Wastes may prevent algal respiration at
night by reducing the DO of water.
E Wastes may stimulate other organisms
at the expense of certain algae.
F Products of waste decomposition may
act as powerful growth stimulants for
certain algae.
II ALGAE AND ORGANIC POLLUTION
A Heavy pollution may tend to limit various
kinds of algae to certain zones in the
affected area.
B These zones are distinguished according
to the degree of change which has
occurred in the organic wastes. One set
of names for these zones includes the
Polysaprobic, alpha-mesosaprobic, beta-
mesosaprobic and oligosaprobic.
C A few "pollution" algae are common in
the first two zones. Many algae are
common in and often limited to one or
both of the last two zones.
D Some workers have listed separately
those algae indicative of each of the four
zones.
Ill REASONS FOR SELECTIVITY OF
POLLUTANTS TO ALGAE
A Certain components of wastes are chemi-
cals toxic to some algae but not to others.
IV ALGAE AS INDICATORS OF POLLUTION
A Selection of list of "pollution" algae
follows an evaluation of the kinds re-
ported in published reports by numerous
workers as relatively prominent in, or
representative of, the polysaprobic and
alpha-mesosaprobic zones in a stream
polluted with sewage. It includes also
other conditions or areas approximating
these zones.
B A total list of more than 1000 kinds of
algae has been compiled to date.
1 In order to tabulate the information,
an arbitrary numerical value is
allotted to each author's record of
each pertinent alga.
2 The algae are then arranged in order
of decreasing emphasis by the
authors as a whole.
BLIND. 10a.8. i
IV 2-1
-------
Algae as Indicators of Pollution
VI SOME GENERA AND SPECIES OF ALGAE
HIGH ON THE LIST ARE AS FOLLOWS:
A Genera: Oscillator la, Euglena, Navicula,
Chlorella, Chlamydomonas, Nitzschia,
Stigeoclonium, Phormidium, Scenedesmus,
Ankistrodesmus. Phacus.
B Species: Euglena viridis, Nitzschia
palea, Oscillatoria chlorina,
Oscillator la limosa, Oscillatoria tenuis
Scenedesmus quadricauda, Stigeoclonium
tenue, Synedra ulna and Pandorina morum.
VII SOME ALGAE REPRESENTATIVE OF
CLEAN WATER ZONES IN STREAMS:
Chrysococcus rufescens, Cocconeis
placentula, Entophysalis lemaniae, and
Rhodomonas lacustris.
VIII RELIABILITY IN USE OF INDICATORS
DEPENDS IN PART UPON ACCURATE
IDENTIFICATION OF SPECIMENS
REPRESENTATIVE LITERATURE
1 Brmley, F. J. Biological Studies. Ohio
River Pollution Survey. I.
Biological Zones in a Polluted Stream.
II. Plankton Algae as Indicators of
the Sanitary Condition of a Stream.
Sewage Works Journal, 14:147-159.
1942.
2 Butcher, R.W. Pollution and
Repurification as Indicated by the
Algae. Fourth International
Congress for Microbiology (held) 1947.
Report of Proceedings. 1949.
3 Fjerdingstad, E. The Microflora of the
River Moelleaa with Special Reference
to the Relation of the Benthal A Igae
to Pollution. Folia Limnological
Scandinavia. No. 5. 1950.
4 Fjerdingstad, E. Taxonomy and Saprobic
Valency of Benthic Phytomicro-
Orgamsms. Intern. Rev. Ges.
Hydrobiol. 50:475-604. 1965.
5 Hawkes, H.A. The Biological Assess-
ment of Pollution in Birmingham
Streams. The Institute of Sewage
Purification, Journal and Proceeding's.
177-186. 1956.
6 Kolkwitz, R. Oekologie der Saprobien.
Schriftenreiche des Vereins fu"r
Wasser-, Boden-, und Lufthygiene
Berlin-Dahlem. Piscator - Verlage,
Stuttgart.
7 Lackey, J. B. The Significance of
Plankton in Relation to the Sanitary
Condition of Streams. Symposium
on Hydrobiology. University of
Wisconsin. 311-328. 1941.
8 Liebmann, H. Handbuch der
Frisbhwasser - und Abwasserbiologie.
R. Oldenbourg, Munchen.
9 Palmer, C.M. Algae as Biological
Indicators of Pollution. In
Biological Problems in Water
Pollution. Trans, of 1956 Seminar.
Robert A. Taft Sanitary Engineering
Center. 1957.
10 Palmer, C.M. The Effect of Pollution
on River Algae. Annal. N. Y. Acad.
Sci. 108:389-395. 1963.
11 Palmer, C.M. A Composite Rating of
Algae Tolerating Organic Pollution.
Jour. Phycology 5 (l):78-82. 1969.
12 Palmer, C.M. Algae in Water Supplies
of the United States. In: Algae an'd
Man.Ch 12, Plenum Press. N.Y.
pp. 239-261. 1964.
13 Patrick, R. Factors Effecting the
Distribution of Diatoms. Botanical
Review. 14: 473-524. 1948.
14 Whipple, G.C., Fair, G.M. and
Whipple, M.C. The Microscopy of
Drinking Water, 4th ed. J. Wiley
and Sons. New York. 1948.
This outline was prepared by C.M. Palmer,
Aquatic Biologist, Cincinnati Water Research
Laboratory, FWPCA.
IV 2-2
-------
Algae as Indicators of Pollution
POLLUTED WATER ALGAE
PHORMIDIUM
PLATE 3
-------
Algae as Indicators of Pollution
CLEAN WATER ALGAE
CLADOPHORA
PLATE 4
-------
PUBLIC HEALTH SIGNIFICANCE OF TOXIC ALGAE
I INTRODUCTION
A Increasing interest is being shown
in toxic algae as a result of a recent
increase in the number of reported
cases of animal poisoning, and sus-
pected cases of direct human poison-
ing thereby. Indirect poisoning of
man from eating poisoned shellfish
has been well demonstrated, and the
poisoning of fish (for human con-
sumption) by algae is suspected.
B Two general groups of algae are of
outstanding importance:
1, The blue green algae (Cyanophy-
ceae).
2 Certain armored flagellates
(Dmophyceae).
3 Although both groups occur in
both fresh and salt waters, the
blue green algae alone are known
to be toxic in fresh water. In
marine waters, the armored
flagellates predominate but
other groups are also involved.
C. The biochemical nature of algae
toxins is not well understood. They
seem in general to be substances of
low molecular weight. They are
capable of producing death or illness
in mammals and fish even when the
algae cells themselves have been
excluded.
II TOXIC FRESH WATER ALGAE
A. Blue green algae associated with
toxic outbreaks include the following
(not all blue green algae are toxic):
TABLE I
Toxic Fresh-Water Algae
Anabaena
Anabaena circmahs
Anabaena flos-aquae
Anabaena lemmermanni
B
Anacystis (Microcystis)
Anacystis cyanea (Microcystis aerugmosa)
Anacystis cyanea (Microcystis flos-aquae)
Anacystis cyanea (Microcystis toxica)
Aphamzomenon flos-aquae
Gloeotrichia echinulata
Gloeotrichia pisum
Gomphosphaena lacustris
(Coelosphaerium kuetzmgianum)
Lyngbya contorta
Nodularia spurnigena
Rivularia fluitans
Instances of animal poisoning from
algae polluted waters are numerous.
1 Toxicity to domestic animals
has been reported from around the
world.
a First published account in
1878 reported sheep, horses,
dogs, and pigs dying in South
Australia after drinking water
containing a heavy concentra-
tion of Nodularia spumigena.
b Arthur 1886 reported cattle
in Minnesota dying within one-
half hour after drinking algae
polluted water.
c Hatch 1959 has reported stock
killed and made ill by drinking
algae polluted water from barn-
yard watering tanks.
d Symptoms generally included
prostration and convulsions,
followed by death.
2 Chickens, turkeys, ducks and
geese have also been killed.
3 Dying and decomposing masses
of Aphanizomenon flos-aquae have
killed a variety of fish under ex-
perimental conditions.
L Most of the outbreaks of algae
poisoning have occurred during
periods of continuous hot weather
when the water had a high organic
content, and when the algae were
BI. MIC.txa.3b.4.70
IV 3-1
-------
Public Health Significance of Toxic Alpae
concentrated at one end of a lake
or pond by a gentle wind.
5 Only a single species of alga has
been involved in each instance.
Until recently, no cases of human
illness or death have been reliably
traced to drinking water containing
toxic algae.
1 During droughts of '30s widespread
gastroenteritis in several eastern
cities was ascribed by some in-
vestigators to huge growths of
algae in rivers used as water
supplies, even though water was
run through water treatment
processes including heavy
chlonnation.
2 Recent reports from Saskatchawan
indicate that heavy blue green algae
growths there have seriously m-
terferred with the recreational
use of water at certain resort
areas.
3 The toxic material from certain
algae may survive the laboratory
equivalent of water treatment using
alum coagulation, filtration and
chlonnation. It may survive ac-
tivated carbon treatment in amounts
ordinarily used in water treatment
plants.
Ill TOXIC MARINE ALGAE
A Toxic marine algae are found pre-
dominately among the armored flag-
ellates. The greens, blue greens,
aiid others also contribute toxic
forms. The following have been
listed as significant:
TABLE II
Toxic Marine Algae
Armored flagellates (Dinoflagellates)
Cochlodimum catenatum
Exuviaella baltica
Gonyaulax catenella
Gonyaulax polyedra
Gonyaulax tamarensis
Gymnodmium brevis
Gymnodinium veneficum
Gymnodmium splendens
Gymnodinium mitimoto
Pyrodmium phoneus
Flagellate
Hornelha marina
Prymnesium parvum
Brown
Egregia laevigata
Hesperophycus harveyanus
JViacrocystis pyrifera
Pelvetia fastigiata
Red
Gelidium cartilagmeum var. robustum
Green
Caulerpa serrulata
Blue Green
Lyngbya aestuarn
Lyngbya majuscula
Trichodesmium erythraeum
B Blooms of planktomic marine algae
may be of various colors, often
called '!Redtide"or "Red water".
1 May or may not be toxic.
2 Toxicity may be due to a true toxic
fraction or to secondary conditions
such as oxygen deficiency, hydro-
gen sulfide liberation on decom-
position, or associated bacterial
pollution.
C Occurrence of Red Tides
1 Trichodesmium - widespread and
recurrent patches in the Philippines,
East IndiAn Archipelogo, and along
the coast of South America.
IV 3-2
-------
Public Health Significance of Toxic Algae
D
2 Tnchodesmium and dinoflageHates -
Red Sea and north-west Indian
Ocean.
3 Chloromonadmeae (Hornellia
marina) occurs off the south-
east and south-west Indian Coast.
4 Chrysophysean flagellate
(Prymnesium paryum) occurs in
Denmark and Israeli ponds.
5 Dinoflagellates blooms associated
with untoward conditions have
been universally recorded from
all over the world.
a Southern California - Gonyaulax
polyedra and G. catenella.
b Washington State - Gymnodmium
splendens.
c Florida and Gulf of Mexico -
Gymnodmium brevis, and
Gonyaulax catenella.
d Bay of Fundy - Gonyaulax ta-
marensis.
e Belgium - Pyrodmium phoneus.
f Portugal - Gonyaulax polyedra.
g Angola, West Africa - Exuyiaella
baltica.
h Japan - Gymnodmium miki-
motoi and Cochlodinium
catenatum.
i Sidney Australia - Gonyaulax
polyedra.
Toxic Dinoflagellates may be present
and cause damage even though not
always present in sufficient numbers
to cause visible red tides.
1 Gonyaulax catenella occurs from
Southern California to Alaska.
It is the primary source of the
poison in shellfish on the West
Coast.
a Organism found in stomachs
of toxic mussels in consid-
erable numbers.
b Yearly maximun of Gonyaulax
occurs during and preceding
the poison in mussles.
c Mussles kept in the laboratory
in clean aerated sea water
lose their toxicity, whereas
if Gonyaulax is present they
increase in toxicity.
d An acid extract of collected
organisms contains the poison.
e Organisms have been grown
in pure culture and shown to
contain the poison.
2 Gonyaulax Tamerensis is believed
to be the cause of poisonous shell-
fish in the Bay of Fundy.
3 Pyrodinium phoneus is believed to
be the cause of poisonous shell-
fish in Belgium.
4 Gymnodmium yeneficum toxin has
been obtained salt-free by dialysis
and concentrated by evaporation
under reduced pressure. Isolated
from an area near Plymouth,
England.
5- Gymnodmium brevis - Bacterial
free cultures with concentrations
from 2. 3 to 4.8 million organisms
per liter were toxic to three
species of fish.
E Physiological Effects
1 Gonyaulax catenella toxin causes
paraesthesia, loss of strength in
mussels of the extremities and
neck, and death by respiratory
failure. Toxin apparently does
not harm shellfish, but shellfish
may concentrate and store the
poison in their tissue. Subsequent
ingestion of the shellfish by man
or other animals may cause death.
IV 3-3
-------
Public Health Significance of Toxic Algae
2, Gymnodmium veneficium toxin
may cause massive fish kills.
Small fish, namely gobies, die
within 10 minutes in toxic
cultures. Action seems to be
on the nervous system and prob-
ably is due to respiratory failure.
Injection of the toxin into the
dosal lymph sac of a frog has an
immediate paralyzing effect.
Ecological factors leading to bloom
are not well understood. It appears
that high concentrations of nutrients,
particularly nitrate and phosphate
must be available. Temperatures
must be suitable and there must be
enough phytoplankton present as an
inoculum to take advantage of these
conditions.
1 Oceanographic factors
a. Water temperature - Gonyaulax
tamerensis has been shown
to be most prevalent in the
Bay of Fundy when the water
temperature is between 10
and 14°C, with an optimum
of 13.9°C.
b Salinity - optimum unknown but
believed to be a factor.
c Nutrients - phosphates, and
Vitamin B12 have been shown
to be essential to dinoflagellate
metabolism.
d Turbulence of water.
1) Upwellings of ocean currents
along the California Coast,
occasionally and unpredic-
tably bring a supply of
critical nutrients to the
s urface from abyssal depths.
2) Tidal turbulence as in the
Bay of Fundy may expose
geologic deposits of nut-
rients.
3) Heavy rains on land have
washed nutrient materials
into the sea and caused
blooms in Florida.
2 Small amounts of sewage pollution
also may favor growth by contri-
buting nutrient material.
3 Associated phenomena
a Extraordinary wide and rapid
variations in the dissolved
oxygen content of the water
have been observed during
and immediately following
heavy concentrations of
Gonyaulax.
b Biochemical oxygen demand
when Gonyaulax are present
is far in excess of what
might be explained by sewage
or other organic pollutants.
c Dinoflagellate blooms are often
succeeded by blooms of or-
ganisms which prey on them.
In the Bay of Fundy the ciliate
protozoa, Fevella, ebrenbergii
was found feeding upon Gony-
aulax.
d Other organisms such as
diatoms may compete with
Gonyaulax for food or light
and check their growth.
Some of the complexities of dino-
flagellate ecology are suggested by
the epidemiology of fish poisoning
suffered by people throughout
tropical regions.
1 It has long been known that some
species of coral are crammed with
zooxanthelae symbiotic algae whose
role in the corals economy is still
obscure.
a The dinoflagellate Gymnodmium
adriaticum has recently been
isolated from a jellyfish, some
sea animones and corals,
G. adriaticum is not toxic but
IV 3-4
-------
Public Health Significance of Toxic Algae
other local species may be.
The parasitic dinoflagellate
Odinum which lives on fish gills
or skin should also not be over-
looked.
Coral animals and shellfish which
feed upon free swimming dino-
flagellates could also pass their
poison up the food chain to fish
and prediators which feed on fish.
Lyngbya and a few other algae also
have been suggested as a cause
of fish poisoning.
It has been suggested that there
are as many as eight different
kinds of fish poisoning, but only
two or three of these have been
studied sufficiently to indicate their
similarity or dissimilarity.
a Puffer fish poisoning - very
similar to shellfish poison but
not identical. Symplons most
frequently develop in 10 to 45
minutes and include paresthesia,
hypersahvation, sub-normal
temperature, decreased blood
pressure and a weak pulse.
The paresthesia gradually
develops into severe numbness
and finally terminates in ex-
tensive muscular paralysis.
If death occurs, it is generally
within 4 to 24 hours and results
from a progressive ascending
paralysis involving the res-
piratory muscles.
b Ciguatera poisoning - barracuda,
snappers and various reef fish
are most commonly incriminated.
The poison is fat soluble and on
this basis appears to be distinct
from shellfish poison or puffer
fish poison. Symptoms develop
in from 1 to 6 hours and include
paresthesia, headache, fever,
profuse sweating, rapid weak
pulse, prostration, muscle
pains, and joint aches. Initial
symptoms sometimes may
consist of nausea, vomiting
and diarrhea, and in severe
intoxications, paradoxical
sensory disturbances may
be present in which hot ob-
jects may feel cold. Death
may occur within a few
minutes but generally re-
quires several days. Mus-
cular twitchings, tremors
and convulsions are followed
by death due to respiratory
failure.
Gymnothorax poisoning -
frequently caused by co^sum -
ing moray eels. As with
ciguatera and puffer poisoning,
the initial symptoms include
tingling and numbness about
the lips, tongue, hands and
feet, sometimes followed by
nausea, vomiting, a metallic
taste, diarrhea and abdominal
pain as in ciguatera. The
characteristic signs, however,
appear to be profuse perspira-
tion, excessive mucus produc-
tion, rapid respiration, high
fever, purposeless movements
and violent convulsions. In
severe intoxication, death does
not occur until after 14 to 25
days.
REFERENCES:
1 Ingram, W. M., and Prescott, G. W.,
"Toxic Fresh-water Algae, " American
Midland Naturalist, 5^:75, 1954.
2 Palmer, C. M., "Algae in Water Supplies, "
U. S. Dept. of Health, Education, and
Welfare, Public Health Service,
Pub. No. 657, 1959.
3 Hatch, Ray D., Personal Communication,
Dept. of Veterinary Medicine, Univer-
sity of Illinois, Urbana, Illinois, 1959.
4 Dillenberg, H. O., and Dehnel, M. K.,
Toxic Waterbloom in Saskatchewan,
1959, Can. Med. Assoc. J. 83_:1151,
19GO.
IV 3-5
-------
Public Health Significance of Toxic Algae
5 Bishop, C.T., Anet, E.F.L. J., and
Gorham, P.R., Isolation and Iden-
tification of the Fast-Death Factor
in Microcystis aeruginosa NRC-1.
Can. J. Biochem. Physiol. 37:453.
1959.
6 McFarren, E.F.. Schafer, M.L.,
Campbell, J. E., and Lewis, K.H.,
"Public Health Significance of
Paralytic Shellfish Poison, " Advances
in Food Research JJ):135, Academic
Press, Inc., New York, 1960.
7 Hutner, S. H., and McLaughlin,
"Poisonous Tides. " Scientific
American, 199:92. 1958.
8 Ballantine, D., Abot, B.C., "Toxic
Marine Flagellates: Their Occurrence
and Physiological Effects on Animals, "
J.Gen. Microbiol. .16:274, 1957.
9 Ray, S.M., and Wilson, W.B., "The
Effects of Unialgal and Bacteria-Free
Cultures of Gvmnodinium brevis on
Fish and Notes on Related Studies
with Bacteria. " Special Scientific
Report - Fisheries No. 211. United
States Department of Interior, Fish
and Wildlife Service, 1957.
10 Habekast, R.C., Fraser, I. M., Halsted,
B.W., "Toxicology - Observations on
Toxic Marine Algae, " J. Washington
Academy Sci. 45_:101, 1955.
11 Banner, A.H., A Dermatitis-Producing
Alga in Hawaii. Hawaii Medical
Journal .19:35, 1959.
12 McFarren, E.F., Report on Collaborative
Studies of the Bioassay for Paralytic
Shellfish Poison, J. Assoc. Offic. Agri.
Chemists 42:263, 1959.
13 McFarren, E.F., and Bartsch, A.F.,
Application of the Paralytic Shellfish
Poison Assay to Poisonous Fishes,
J. Assoc. Offic. Agri. Chemists
43:548, 1960.
14 Ulitzur, S. , Purification and Separation
of the Toxins Produced by the Phyto-
flagellate Frymnesium parvum. Verh.
Internat. Verein. Limnol. 17:771-777.
1969.
15 Jackson, Daniel F. , Algae, Man, and the
Environment. Syracuse Univ. Press.
554 pp. 1968.
This outline was prepared by H.W. Jackson,
Chief Biologist, National Training Center,
FWPCA, Cincinnati, OH 45226. Portions
of this outline were taken from previous
training outlines by W. M. Ingram and
E. F. McFarren.
IV 3-6
-------
ORGANIC ENRICHMENT AND'DISSOLVED OXYGEN RELATIONSHIPS
IN WATER
I INTRODUCTION
A Oxygen normally composes approximately
20% of the atmosphere, but from only 0. 5
to 1% of unpolluted water by volume
(9 - 14 milligrams per liter, or parts per
million).
B Physically, oxygen is a colorless gas,
responsive to the laws of solubility,
temperature, etc.
C The amount of oxygen present can be
readily determined by various methods,
both chemical and physical.
D Biologically, oxygen is one of the key
elements to all life, all living organisms
contain it along with hydrogen and carbon.
It is the physiological "complement" of
carbon dioxide.
E When properly interpreted, oxygen con-
centration can serve as an excellent index
to water quality.
1 It is widely present.
2 It is intimately involved in many
processes, chemical, physical, and
biological.
H BIOLOGICAL OR PHYSIOLOGICAL
RELATIONSHIPS (Figure 1)
A Photosynthesis is the biological starting
point for synthesizing living substance out
of non-living carbon dioxide, water, and
other materials. In the process, radiant
energy is adsorbed from the sun (hence it
is an endothermic reaction) and free
molecular oxygen is released to the environ-
ment as long as sufficient light is available,
i. e., during daylight only.
1 The conventional empirical formula for
photosynthesis is a very great sim-
plification, as many as twenty steps
may actually be involved (like
respiration, controlled by systems
of enzymes).
6 CO_ + 6 H,0 + 673 Cal
£ &
(Energy sunlight)
chlorophyll
(enzyme system)
C6H12°6+6°2
(Simple sugar)
The end product is a basic organic
material known as a simple sugar
(carbohydrate, monosaccharide, of
which "glucose" is an example,
see Figures 1, A) which serves as a
starting point for the elaboration of
higher organic substances, and which
may also be oxidized or "respired" by
the cell to release the stored chemical
energy.
Synthesis of organic materials (living
protoplasm) from simple sugars and
other substances in bacteria, algae,
and man.
a Glucose and other simple sugar
molecules can be combined by
living organisms such as algae into
other carbohydrates such as starch.
b After modification into fatty acids
and glycerol, fats can be made.
c Combination with nitrogen and
other minerals taken in from the
surrounding water produces amino
acids. These can then be combined
into proteins, the most complicated
of aU life substances. Plants are
the only organisms that can synthesize
amino acids completely "from scratch. "
BI. ECO. 23.5. 70
IV 5-1
-------
Organic Enrichment and Dissolved Oxygen Relationships in Water
ENERGY CYCLES
H W Jackson
A . THE GENERAL FOOD CYCLE
mineral* in the protoplasm
of algae and other plant*
carbohydrates
fate
protein*
S
k r1
mineral* in
microcruitacea.
iniect larvae,
other animal*
a
ABS
A
PHOTO1,
CHEMICA
FIXATION
Mineral* in
forage fi*h
i
mineral* in
predator*
ANIMAL
EXCRETION
coz
H20
other
mineral*
P
K
N0j
ANIMAL AND
PLANT
RESPIRATION
BACTERIAI
lESPIRATIOr
mineral* in
bacterial
protoplasm
a
B. SYNTHESIS AND DEGRADATION OF PROTOPLASM
THE ENVIRONMENT
3
5
Iengymea of photosynthesU (chlorophyll)
^ enayrne* ol respiration
Carbohydrate*: poly*accharlde*_ dlsaccharides** monosaccharide*
Fat*: fat* and llpid*s= fatty acid* and glycerol ap=i-J |
Protein.: protein.* feStaSEHr
\ i S
urn and other mineral -
containing excreta
combined with N
and other
mineral*
BI.ECO. 2. 11.55
Figure 1
IV 5-2
-------
Organic Enrichment and Dissolved Oxygen Relationships in Water
TS£ S&eOK-SXYfiEN CYCLE m
CARBON DIOXIEtt ,
RESPIRATION
PHOTOSYNTHESIS
E1EMY for L
PROCESSES
ORGANIC MATERIAL DESTROYED
(OXYGEN CONSUMED)
ORGANIC MATERIAL, CONSTRUCTED
(OXYGEN RELEASE!?
ecoto«H,o«
BI.EC0.2a.3.58
Figure 2
4 All of these synthetic or growth
processes are endothermic and require
energy to consummate. This energy
is obtained by respiration.
Respiration. Life processes such as those
mentioned previously (II A 3, also physical
movement) are energized by the controlled
oxidation or "respiration ' of single sugars.
This is a cellular or physiological process
and continues constantly in all living things,
independently of light, as long as life
persists.
1 Types of respiration. Respiration is
an energy releasing or exothermic
biochemical process and is to be
distinguished from the external bodily
act of "breathing", which is a process
involving a lung, gill, or other organ,
whereby molecular oxygen is brought
inside the body of the organism and
carbon dioxide is released. Breathing
is transportation; respiration is
burning or biochemical utilization.
a Aerobic respiration. The most
common form of respiration is
illustrated by the oxidation of
glucose, using free dissolved
oxygen from the environment.
This is also the most efficient
type since virtually all of the
chemical energy contained in the
glucose molecule is released
(673 Cal.) and the residue is com-
pletely mineralized or stabilized to
carbon dioxide and water.
-------
Organic Enrichment and Dissolved Oxygen Relationships in Water
Although actually taking place in
more than twenty steps involving
as many different enzymes, the
overall process can be empirically
represented by the following
formula:
C6H12°6
6°
(a simple sugar such as glucose)
6 CO2 + 6 H2O + 673 Cal
(enzymes)
(energy)
b Anaerobic respiration. If free
dissolved oxygen (DO) is not available
from the environment, certain types
of bacteria, fungi, and other orga-
nisms known as "facultative aerobes"
can switch to another, less efficient
type of respiration and still obtain
energy by breaking down simple
sugars. This is known as anaerobic
respiration. Some types of micro-
organisms, the true anaerobes, can
thrive only under anaerobic conditions.
The anaerobic respiration of glucose
involves splitting the molecule with-
out the use of outside oxygen so as
to release some of the contained
energy, but not all. The remainder
is still available in the end products.
A common empirical relationship is
as follows:
3 C3H12°6-
2 CH3(COOH)2
(glucose) (enzymes) (lactic acid)
+ 2 CH_CH0OH + 2 CH, COOH
J Ct O
(ethyl alcohol)
+ 59 1/2 Cal
(energy)
(acetic acid)
c Anaerobic respiration of proteins
and their derivations by micro-
organisms (known as putrefaction)
often leads to the production of foul
smelling end products. Aerobic
respiration of proteins (known as
decay) seldom produces offensive
odors. In addition to carbohydrates,
proteins, fats, and various other
natural and unnatural hydrocarbons,
and other substances can also be
broken down or respired to rel-
atively stable end products by
microorganisms.
The adaptability of life is remarkable.
By the same mechanisms of genetics
and natural selection which have
resulted in the evolution of present
day life on earth, microorganisms
have evolved which are capable of
obtaining energy from, or in other
words, "respiring" a great variety
of new synthetic materials, the products
of modern industry. This is the basis
for the biological treatment of wastes.
a The types of biological mechanisms
involved, in general, are known
and to a degree, understood.
b Many species or kinds of orga-
nisms may be involved in the
stabilization of a single type of
waste.
Significance of respiration in sanitary
engineering. The process of respi-
ration uses or "demands" oxygen from
the environment. Any material, such
as sewage, an organic industrial waste,
motor oil, etc., which will support
life of any kind will thus exert (through
the organisms living on it) a "BOD".
a Respiration has two end "products":
energy, by which the organisms
live; and degraded, mineralized,
or stabilized material such as
digested sewage sludge which
retains little or no energy available
to organisms. That is, they have
no biochemical oxygen demand or
BOD remaining.
IV 5-4
-------
Organic Enrichment and Dissolved Oxygen Relationships in Water
Since it is independent of light, it
goes on night and day.
IE PHYSICAL PHENOMENA
A Solubility of oxygen in pure water varies
with temperature and pressure according
to well known laws and values.
1 We expect to find less oxygen in warmer
waters and more in colder. This has
both seasonal and geographical
implications.
2 Dissolved substances such as salts tend
to reduce the solubility of oxygen at a
given temperature (See Figure 3 -
Oxygen Solubility at Selected Salinities).
T B 9 IO M 12 13 14
DISSOLVED OXYGEN pern
OXYGEN SOLU9ILITY AT SELECTED SALINITIES
Figure 3
The concentration of oxygen often is
expressed as percent of saturation.
a Organisms, however, are affected
by the real quantities present, and
actually demand the most oxygen at
the highest temperature, when the
solubility is lowest.
b It is therefore more significant,
where organisms and stream con-
ditions are concerned, to express
concentration as milligrams per
liter (or parts per million).
B The movement or distribution of oxygen
throughout a water mass is not entirely
dependent on molecular diffusion, but
also involves various types of gross
water movements. Reaeration (or aeration)
is the transfer of oxygen from the air into
the water mass.
IV CHEMICAL FACTORS
A Chemical toxicity may delay the exertion
of biological oxygen demands, or the
release of oxygen by biological mechanisms.
B Chemical substances may themselves
demand oxygen from the water.
1 If oxygen is available, the chemical
oxygen demands tend to be satisfied
relatively quickly and hence are likely
to be a more local problem.
2 They are independent of any biological
effect.
3 They are in addition to biological
action.
V OXYGEN AND EUTROPfflCATION
A Diurnal interactions of photosynthesis
and respiration. (Figure 6)
1 Life, growth, and hence the need for
oxygen for respiration continue
twenty-four hours a day, as long as
life persists.
2 Since photosynthesis in nature is
activated by radiant energy from the
sun, it is operative only during the
daytime. It releases approximately
twenty times as much oxygen as is
consumed in cellular respiration,
however, so there is a great excess
left over which diffuses out into the
surrounding water. Here it is available
to biochemical oxidative demands of
other organisms with the result that
the ecological system becomes "aerobic. "
IV 5-5
-------
Organic Enrichment and Dissolved Oxygen Relationships in Water
3 DO's affected by photosynthesis thus
tend to be highest in the daytime
(particularly mid-afternoon) and lowest
at night (3 - 4A.M.).
B Organic materials from domestic sewage
or similar wastes are readily available
to microorganisms. Assimilation of food,
and growth and multiplication of the pop-
ulation can thus begin relatively quickly,
thus establishing a demand for oxygen for
respiration.
1 It takes a significant period of time for
a population of bacteria, fungi, and
other microorganisms such as protozoa
to become established, after the
initiation of growth of the few original
organisms, even under the best of
conditions.
2 Dissolved oxygen does not thus disappear
instantly from the water upon the
admixture of sewage, but rather begins
to diminish slowly. As the population of
oxygen consuming organisms builds up,
the deficit in the water increases. As
this occurs, any equilibrium which may
have existed with the atmosphere is
destroyed, and the rate of aeration
through the surface film increases.
a As the population of microorganisms
grows still larger, its rate of increase
begins to increase (logarithmic
growth). Very soon a size of pop-
ulation is achieved where the demand
for oxygen exceeds the amount
which can be supplied through the
surface film. The concentration of
free DO now begins to drop sharply
and may go to zero.
b If sufficient food for the micro-
organisms remains, anaerobic
conditions will now prevail.
Oxygen, of course, continues to
enter at a maximum' rate through
the surface film, but is immediately
used up. The overall rate of
oxidation of the waste or food
material however, is now very low
(although the total amount may be
large), as surface aeration can
supply but a small portion of the
oxygen needed.
If there is no replenishment of the
food supply by repollution, the big
population of fungi and bacteria
eventually uses up most of the
available substrate or food, and
begins to starve. Growth now
begins to slow down, and the pop-
ulation eventually drops back to
its original low level.
Meanwhile time has passed. An
abundance of fundamental plant
nutrients such as nitrate, phosphate,
potash, etc., have been released
to the water as a result of micro -
bial respiration. Algae begin to
grow on this substrate, and increase
rapidly to tremendous numbers.
Algal photosynthesis thus suddenly
begins to release great quantities
of oxygen in the daytime. At first
this is exhausted each night by
respiration of the algae and other
microorganisms, but soon persists
around the clock until the algae in
turn have exhausted their food
supply and return to numbers
normal for an unpolluted stream.
With the above mentioned advent
of free DO from the algae, the more
efficient aerobic type of respiration
can again be employed by the
microorganisms. This hastens
the oxidation or stabilization process,
and leaves behind but a minimum
residue of well mineralized material
to accumulate on the bottom.
It should be mentioned that turbidity
from suspended microorganisms
and other organic solids frequently
inhibit the establishment of an algal
population until the biochemical
oxidation or stabilization process
has been well started. Turbidity
inhibits algal growth primarily by
shading and suppression of photo-
synthesis.
IV 5-6
-------
Organic Enrichment and Dissolved Oxygen Relationships in Water
When organic material is dumped into a
body of water faster than it can be oxidized
by the mechanism described above, it
tends to accumulate on the bottom in a
partially digested condition known as
sludge.
1 Great quantities of nutrient energy are
still present, as well as a huge popu-
lation of microorganisms. There is a
very great need for oxygen which, if
available, is avidly taken up by the
organisms in the surface layers.
a Aerobic conditions in the overlying
waters thus tend to hasten the
stabilization of sludge banks by
providing oxygen for aerobic type
respiration.
b Anaerobic waters, on the other hand,
tend to preserve the sludge by
restricting all organisms to anaer-
obic types of respiration. It has
been reported that the speed of
decomposition begins to be restricted
at concentrations below 1 ppm.
Figure 4
Non-Pigmented, Non-Oxygen-Producing,
Protozoan Flagellates
2 Anaerobic conditions are present
throughout the sludge mass below the
surface. Stabilization is still
proceeding, but at a much slower
rate than if oxygen were present.
D In the deeper reservoirs, lakes, rivers,
and estuaries thermal stratification may
develop in the summer.
1 The epilimnion or upper layer, being
in contact with the atmosphere,
receives a continual replenishment of
oxygen by aeration processes.
2 If turbidity is not excessive, photo-
synthesis by algae (Figure 5) will
also provide oxygen.
3 The hypolimnion or lower layer, being
separated from the atmosphere and
frequently from the light, tends to
acquire an excess of carbon dioxide
and a deficiency of oxygen due to the
respiratory activities of micro-
organisms (such as protozoa, fungi,
and bacteria). (Figure 4)
This effect is heightened if the bottom
material is high in organic content
as has been shown below.
Figure 5
Pigmented, Oxygen-Producing, Algal
Flagellates
7
-------
Organic Enrichment and Dissolved Oxygen Relationships in Water
VI PROBLEMS AND BENEFITS RESULTING
FROM ALGAE PRODUCTION
(Figures 4, 5, 6)
A Problems to man may result when the total
"primary production" by algae leads to an
increase in the total organic content of the
water that interferes with a desired use.
Death of large algal populations may
lead to obnoxious odors through
bacterial decomposition. Oxygen
deficits may result at any time of day
in this process. Deposition of masses
of organic sediment or sludge in
estuaries and back waters may be
considerable.
This may consist of a high algal
population that produces a water with
high turbidity, taste and odor, or other
undesirable effect. High respiratory
needs may lead to nocturnal oxygen
deficit.
Certain algae may cause tastes and
odors, clog filters; or otherwise inter-
fere with potable water processing.
B The primary production of algae can also
serve as a supply of food to consumer
organisms (animals), resulting in
increased production at several (trophic)
levels: (Figure 7)
zobmicrobes, microinvertebrates,
macroinvertebrates, fishes.
9.0
(OCTOBER 2, 1957)
e.o
2
i
0
I i.o
4 o.s
0.0
(OCTOBER 3, I9S7)
SOUR RADIATION, MAM CALORIES/
SO,. CM./MIMUTE
RESPIRATORY USE AND 0.0. PRODUCTION
AT THREE DEPTHS
O.S FT.
MEAN
RESPIRATION
DISSOLVED OXYGEN RELATIONS IN OHIO RIVER (REACH II, BROMLEY)
OCT. 3, 1067
Figure 6
IV 5-8
-------
Organic Enrichment and Dissolved Oxygen Relationships in Water
Mean Surface Water Productivity Rates of the Great lakes
and Other Freshwater Lakes and Ocean Areas.
Lake or Ocean
and Location
Lake Superior
Lake Hvron
Lake Michigan
Lake Erie
Grand Traverse Bay,
N.E. Lake Michigan
Douglas Lake, Mich.
Bras d'Or Lake,
Nova Scotia, Canada
Brooks Lake, Alaska
Maknck Lake, Alaska
Torne Trask, Sweden
Ransaren, Sweden
Lake Erken, Sweden
Lake Esrom, Denmark
Lake Purest, Denmark
N.E. Atlantic -
near Denmark
North' Atlantic, same
lat. as Great Lakes
Southeast Pacific
Southwest Pacific
North Pacific,
Sub-Arctic Region
Productivity Rate
in mg.C/ m^ /day
16.62
23.04
37.62
175.20
0.34
0.23
23.5
3.2
8.8
6.4
8.8
221.0
1600
2100
21
7.5
3.3
7.2
6.4
Investigator
Parkos (1967-68)
Parkos (1968)
Parkos (19'67)
Parkos (1968)
Saunders e± al. (1962)
Saunders et_ al,. (1962)
Geen and Hargrave (1966)
Goldnan (1960)
Goldman (1960)
Rodhe (1958b)
Rodhe (1958b)
Rodhe (1958b)
Jonasson and
Mathiesen (1959)
Jonasson and
Mathiesen (1959)
Steemann-Nielsen (1960)
Steemann-Nielsen (1958b)
Holmes (1961)
Angot (1961)
Koblentz-Mishke (1961)
Figure 7
-------
Organic Enrichment and Dissolved Oxygen Relationships in Water
1 Earlier notation cited the release of
oxygen during utilization of CO9 during
algal photosynthesis. This encourages
fungal or bacterial breakdown of
pollutants.
2 Photosynthesis occurs in the presence
of adequate light and favorable conditions.
In darkness, the cells continue to
respire and may consume more oxygen
than they produced because photo-
synthesis increases the organic load.
3 Photosynthesis tends to occur at the
surface where light intensity is greatest
(Figure 6). Poor vertical mixing
would result in stratification of water
supersaturated with oxygen over oxygen
deficient water. Depending upon con-
ditions, a significant fraction of the
oxygen could be lost to the atmosphere.
ACKNOWLEDGEMENTS:
This outline contains certain material sub-
mitted by F. J. Ludzack and M. E. Bender.
REFERENCES
1 Anonymous. Aquatic Life Water Quality
Criteria. Aquatic Life Advisory
Committee on the Ohio River Valley
Water Sanitation Commission. Second
Progress Report. Sewage and Ind.
Wastes. 28(5):678-690. 1956.
2 Anonymous. Oxygen Relationships in
Streams. Proc. of Sem. at R. A.
Taft Sanitary Engineering Center.
October 30 - November 1, 1957.
3 Bartsch, A.F. Algae in Relation to
Oxidation Processes in Natural Waters.
Special Publ. No. 2. Ecology of Algae.
Pymatuning Lab. of Field Biology.
U. of Pittsburgh. Pittsburgh, Pa.
pp. 56-71.
4 Carpenter, J.H., Pritchard, D.W., and
Whaley, R.C. Observations of
Eutrophication and Nutrient Cycles in
Some Coastal Plain Estuaries, in:
Eutrophication: Causes, Consequences,
Correctives; pp. 210-221. Proc. of
Symposium. June 11-15, 1967.
Publ. Nat. A cad. Sci. Washington,
DC. 1969.
5 Custer, S.W. and Krutchkoff, R.G.
Stochastic Model for BOD and DO
in Estuaries. ASCE. Jour. San.
Eng. Div. Vol. 95. pp. 865-886.
1969.
6 Ketchum, B.H. Eutrophication of
Estuaries, in Eutrophication:
Causes, Consequences, Correctives;
pp. 197-209. Proc. of Symposium.
June 11-15, 1967. Publ. Nat. A cad.
Sci. Washington, DC. 1969.
7 Olsen, Theodore A. Some Observations
on the Interrelationship of Sunlight,
Aquatic Plant Life, and Fishes.
62nd Annual Meeting of the Am. Fish
Soc. Baltimore. 1932.
8 Richards, F.A., and Corwin, N. Some
Oceanographic Applications of Recent
Determinations of the Solubility of
Oxygen in Sea Water. Limnology and
Oceanography. l(4):263-267.
October 1956.
9 Ruttner, Franz. (Translated by D. G.
Frey and F. E. J. Fry.) Fundamentals
of Limnology. Univ. of Toronto Press.
pp. 1-242. 1951.
10 Trues dale, G. A., Downing, Al, and
Lowden, G. F. The Solubility of
Oxygen in Pure Water and Sea Water.
J. Appl. Chem. 5(2):53-62. 1955.
This outline was prepared by H.W. Jackson,
Chief Biologist, National Training Center,
FWPCA, Cincinnati, OH 45226.
IV 5-10
-------
PLANKTON IN OLIGOTROPHIC LAKES
I INTRODUCTION
The term oligotrophic was taken from the
Greek words oligos -- small and trophein --
to nourish, meaning poor in nutrients.
Lakes with low nutrient levels have low
standing crops of plankton. The term is now
commonly applied to any water which has a
low productivity, regardless of the reason.
II PHYSICA L A ND CHEMICA L CHA RA CTER-
ISTICS OF OLIGOTROPHIC LAKES*
A Very deep; high volume to surface ratio
B Thermal stratification common; volume
of the hypolimnium large compared to the
volume of the epilimnion
C Maximum surface temperature rarely
greater than 15° C
D Low concentrations of dissolved minerals
and organic matter.
1 Phosphorus, less than 1 microgram
per liter
2 NO -Nitrogen, less than 200 micrograms
per liter
E Dissolved oxygen near saturation from
surface to bottom
F Water very transparent, Secchi disk
readings of 20-40 meters are common
G Color dark blue, blue-green, or green
III PLANKTON
A Quantity
1 Standing crop very low
a Ash-free weight of plankton, less
than 0.1 mg per liter (compared to
1 mg per liter or more in eutrophic
lakes).
b Chlorophyll, 1 mg per M or less
c Cells counts, less than 500 per ml
2 Zooplankton to phytoplankton volume
ratio, 19:1.
B Quality
1 European biologists have found
oligotrophic lakes to be dominated by
Chlorophyta (usually desmids),
chrysophyta (such as Dinobryon). and
Diatomaceae (Cyclotella and Tabellaria).
Eutrophic lakes are dominated by
Synedea, Fragilaria, A sterionella,
Melosira, blue-green algae, Ceratium,
and Pediastrum. Nygaard devised
several phytoplankton quotients based
on these relationships
a Simple quotient
Number of species of
Chlorococcales _ if <1, oligotrophic
Desmidiaceae " if > 1, eutrophic
b Compound index
Myxophyceae+Chlorococcales+Centrales+Eugleniaceac
Desmidiaceae
if2.5, eutrophic
c Diatom quotient
Centrales _ if 0-0.2, oligotrophic
Pennales "if 0.2-3.0, eutrophic
BI.ECO.mic.2. 10.66
IV 6-1
-------
Plankton in Oligotrophic Lakes
2 Several lists of trophic indicators have
been published:
Two are listed here
Teiling.
Swedish Lakes
Rawson,
Canadian Lakes
Oligotrophic Tabelleria flocculosa
Dactylococcopsis
ellipsoideus
Mesotrophic Kirchneriella lunar is
Tetraeadon spp.
Pediastrum spp.
Fragilaria crotonensis
Attheya zachariasii
Melosira granulata
Oligotrophic Asterionella formosa
Eutrophic
Pronounced
Eutrophy
Aphanizomenon spp.
Anabaena flos-aquae
Anabaena circinalis
Microcystis aeruginosa
Microcystis viridis
Mesotrophic
Melosira islandica*
Tabellaria fenestrata
Tabellaria flocculosa
Dinobryon divergens
Fragilaria capucina
Stephanodiscus niagarae
Staurastrum spp.
Melosira granulata
Fragilaria crotonensis
Ceratium hirundinella
Pediastrum boryanum
Pediastrum duplex
Coelosphaerium
naegelianum
Anabaena spp.
Aphanizomenon flos-aquae
Microcystis aeruginosa
Eutrophic
Microcystis flos-aquae
IV 6-2
-------
Plankton ii Oligotrophic Lakes
Some discrepancies can be seen in the
ranking of species in the lists. These
may be the result of true differences in
the composition of the plankton, or may
be only apparent differences which
resulted from different sampling methods.
Many studies (e.g. those by Milliard,
Olive, and Rawson) have been based on
netted samples, which may be highly
biased because they contain little of the
nannoplankton. Also, it is not uncommon
to characterize populations on the basis
of one or two samples collected during
the summer months.
The dominant plankton in four
oligotrophic North American lakes are
listed below. The Great Slave Lake
and Karluk Lake data are from netted
samples taken during the summer, and
monthly, respectively. The Lake
Superior and Lake Tahoe data are from
grab samples taken twice monthly, and
quarterly, respectively.
The dominant diatoms are generally
similar in the four lakes. Asterionella
formosa and Fragilaria crotonensis
are common to all. There are also
some obvious differences. Melosira
islandica, the dominant diatom in the
Great Slave Lake and Lake Superior,
is absent from Lake Tahoe and Karluk
Lake. It was not found in Crater Lake
by Sovereign (1958), in the Mountain
lakes of Colorado by Olive (1955) or
Brinley (1950), and does not occur in
WPSS samples in streams west of the
Great Lakes. Tabellaria is also
absent from Lake Tahoe. It was
reported in Colorado lakes by Olive,
but was not abundant. Brinley makes
no reference to it, and Sovereign
indicated that it was rare in Crater
Lake samples. It is apparent that the
absence of these two diatoms from
Lake Tahoe is not related to the lake.
Except for the absence of Keratella
cochlearis from Lake Tahoe, the
rotifer populations are very similar.
Data on other segments of the zoo-
plankton population are insufficient to
permit comparison.
IV 6-3
-------
en
i
Dominant
Phytoplankton
Dominant
Zooplankton
Raw son.
Great Slave Lake
USPHS,
Lake Superior
Milliard,
Karluk Lake
WPSS,
Lake Tahoe
Melosira islandica
Asterionella formosa
Dmobryon divergens
Ceratium hirundmella
Pediastrum boryanum
Tabellaria fenestrata
Cyclotella meneghiniana
Fragilana crotonensis
Fragilaria capucina
Synedia ulna
Eunotia lunar is
Keratella cochlea r is
Kellicottia longispina
Diaptomus tenuicaudatus
Limnocalanus macrurus
Senecella calanoides
Daphnia longispina
Bosmina obtusirostns
Melosira islandica
Tabellaria fenestrata
Cyclotella kutzmgiana
Melosira granulata
Melosira ambigua
Astenonella formosa
Synedra nana
Scenedesmus spp
Ankistrodesmus spp.
Dictyosphaerium spp.
Keratella cochlearis
Kellicottia longispina
Asterionella formosa
Tabellaria flocculosa
Fragilaria crotonensis
Cyclotella bodanica
Cymbella turgida
Dictyosphaerium spp.
Sphaerocystis spp
Staurastrum spp.
Not reported
Fragilaria crotonensis
Synedra nana
Fragilaria construens
Fragilaria pinnata
Nitzschia acicularis
Asterionella formosa
Kellicottia longispina
Daphnia spp.
Diaptomus tyrelli
Epischura nevadensis
O
B'
O
F.
8
rt-
n
o
"S-
5T
fl>
CD
-------
Plankton in OLgotrophic Lakes
REFERENCES
1 Brinley, F.J. 1950. Plankton population
of certain lakes and streams in the
Rocky Mountain National Park,
Colorado. Ohio J. Sci. 50:243-250.
2 Billiard, O.K., 1959. Notes on the
phytoplankton of Karluk Lake, Kodiak
Island, Alaska. Canadian Field-
Naturalist 43:135-143.
3 Jarnefelt, H., 1952. Plankton als
Indikator der Trophiegruppen der seen.
Ann. Acad. Sci. Fennicae A.IV:l-29.
4 Knudson, B.M., 1955. The distribution of
Tabellaria in the English Lake District.
Proc. Int. Assoc. Limnol. 12:216-218.
5 Nygaard, G., 1949. Hydrobiological studies
in 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.
6 Olive, J.R., 1955. Some aspects of
plankton associations in the high
mountains lakes of Colorado. Proc.
Int. Assoc. Limnol. 12:425-435.
7 Rawson, D.S., 1953. The standing crop
of net plankton in lakes. J. Fish. Res.
Bd. Can. 10:224-237.
8 Rawson, D. S., 1956. The net plankton of
Great Slave Lake. J. Fish. Res. Bd.
Can. 13:53-127.
9 Rawson, D. S., 1956. Algal indicators
of trophic lake types. Limnol.
Oceanogr. 1:18-25.
10 Rodhe, W., 1948. Environmental
requirements of fresh-water plankton
algae. Symb. Bot. Upsal. 10:1-149.
11 Ruttner, F., 1953. Fundamentals of
Limnology, 2nd ed., Univ. Toronto
Press, Toronto.
12 Sovereign, H.E., 1958. The diatoms of
Crater Lake, Oregon. Trans. Amer.
Microsc. Soc. 77:96-134.
13 Telling, E., 1955. Some mesotrophic
phytoplankton indicators. Proc. Int.
Assoc. Limnol. 12:212-215.
14 USPHS, 1962. National Water Quality
Network, Annual Compilation of Data,
PHS Publ. No. 663.
15 Welch, P.S., 1952. Limnology, 2nded.,
McGraw Hill Book Co., New York.
This outline was prepared by C.I. Weber,
Chief, Biological Methods Section,
Analytical Quality Control Laboratory,
FWPCA, 1014 Broadway, Cincinnati, OH
45202.
IV 6-5
-------
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 mineralssalts, 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 in 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 in phytoplankton abundance
may result in taste and odor problems
in water supplies, filter clogging,
high turbidity, changes in 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
IV 7-1
-------
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 islandica var. helvetica
appeared
4 1907 - Stephanodiscus hantzschii
appeared
5 1911 - Bosmina longirostris replaced
B. coregoni
6 1920
1924 - O. rubescens occurred in great
quantities
7 1920 - milky-water phenomenon;
precipitation of CaCCL crystals (40n)
due to pH increase resulting from
photosynthesis
8 Trout and whitefish replaced by perch,
bass, and rough fish
B Hallwilersee, Switzerland
1 1897 - Oscillataria 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 Asterionella appeared, followed
by Synedra
3 About 200 years ago, Asterionella
again became abundant
4 Asterionella abundance ascribed to
domestic wastes
D Finnish Lakes
Aphanizomenon, Coelosphaerium,
Anabaena, Microcystis, are the most
common indication of eutrophy.
TABLE 1 CHANGES IN PHYSIO-CHEMICAL PARAMETERS
Zurichsee, Switzerland
Parameter
Chlorides
Dissolved organics
Date
1888
1916
1888
1914
Value
1.3mg/l
4.9 mg/1
9.0 mg/1
20.0 mg/1
Secchi Disk
before 1910
1905 - 1910
1914 - 1928
Dissolved oxygen, at 1910 - 1930
100 M, mid-summer 1930 - 1942
Max.
16. 8M
10. OM
10. OM
Minimum
it
Min.
3.1M
2.1M
1.4M
100% saturation
9% saturation
IV 7-2
-------
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 organics
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 in 1967.
2 During the period 1965-1967 the Chicago
water treatment plant has found it
necessary to increase the carbon dosage
from 23 Ibs/mil gal to 43 Ibs/mil gal,
and the chlorine dosage from 20 Ibs/mil
gal to 25 Ibs/mil gal.
Phytoplankton counts in the south end
now exceed 10, 000/ml during the
spring bloom.
Ill 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. Raw son 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
IV 7-3
-------
The Effects .of Pollution on Lakes
TABLE 2 PARAMETERS COMMONLY USED TO DESCRIBE CONDITIONS
Oligotrophic Condition
> 10 meters
< lug/1
< 200 ng/1
near 100% saturation
< 1 mg/m
< 0.1 mg/1
< 500/ml
1 Transparency
2 Phosphorus
3 NO, - Nitrogen
0
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 <1
number of species of Desmids
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.
IV 7-4
-------
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,
are:
1 Lake Washington. Seattle
The natural water supply for this lake
is nutrient poor
(Ca = 8mg/l. P< 5Mg/l. 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
(Mg/D
70
66
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 oligotrophic.
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
Eds.
1 Ayers, J. C. and Chandler, D. C.,
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.
l(l):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.
IV 7-5
-------
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
glomerata 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. A cad. 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,
FWPCA, 1014 Broadway, Cincinnati, OH.
IV 7-6
-------
Copper Sulfate
Applicator
CHAPTER V
PLANKTON CONTROL
Control of Plankton in Surface Waters
Control of Interference Organisms in Water Supplies
Nutrients: The Basis of Productivity
3
4
5
-------
CONTROL OF PLANKTON IN SURFACE WATERS
I PHILOSOPHICAL CONSIDERATIONS
A Plankton growths are as natural to aquatic
areas as green plants are to land areas
and respond to the same stimuli.
B Man is currently harnessing plankton forms
to accomplish useful work.
1 For generation of oxygen
a Stabilization of waste waters in
oxidation ponds
b Oxygen recovery from CO2 in space
travel
2 For augmentation of food supply
a Fish ponds
b Nitrogen fixation in rice growing
c Harvesting of algae for direct use
as food
A growing knowledge of the nutrient re-
quirements of plankton organisms will
lead to a more enlightened approach to
ways and means of controlling their growth
when desirable.
II CLASSICAL METHODS OF CONTROL
A Chemical
1 Inorganic
a Copper sulfate is used most exten-
sively. It is most effective in pre-
ventive rather than curative treat-
ment. It has long lasting effects in
soft waters but is short-lived in hard
waters due to precipitation of the
Cu++ as a basic carbonate The pre-
cipitated material accumulates in
bottom muds and is toxic to certain
benthal forms, some of which serve
as important fish food.
Dosages are normally based on the
alkalinity of the water. When alka-
linity is < 40 mg/1, the recommended
dosage is 0. 3 mg/1 of CuSO4 5H2O
in total volume of water. When
alkalinity is > 40 mg/1, recommended
dosage is 2. 0 mg/1 in surface foot
of water.
b Chlorine is preferable to copper
sulfate in the control of certain
forms of algae. However, it is
difficult to apply in most instances
and is very short-lived due to photo
catalytic decomposition of HC1O
HC1 + O
Organic - Numerous organic compounds
have been evaluated, especially in re-
lation to control of blue-green algae.
"Phygon", 2, 3-dichloronaphthoquinone,
has been field tested but is too specific
in its action for general application.
Ill ECOLOGICAL CONTROL
A Theory - Ecological control is based upon
the principle of preventing or restricting
growth by limiting one or more of the
essential requirements. This is an ap-
plication of Liebig's Law of the Minimum.
The logical avenues of control are as
follows:
1 Elimination of light
2 Limiting nutrient materials
B Light - Many cities have solved the prob-
lem of plankton growths by the use of
covered reservoirs, underground and
elevated. Concurrently, they have solved
contamination problems created by birds
and atmospheric fallout In open reservoirs.
BI. MIC. con. lOb. 4.70
V 3-1
-------
Control of Plankton in Surface Waters
some success has been obtained by
limiting light through the use of a film of
activated carbon.
C Nutrients - Since phytoplankton (algae)
serve as the base of the food chain, know-
ledge concerning their nutrient require-
ments is required for ecological control,
when limitation of light is impractical.
The nutrient requirements of phytoplankton
are as follows:
1 Nature of - The major nutrients are:
a Carbon dioxide
b Nitrogen - ammonia and nitrates
(also N2)
c Phosphorus - phosphates.
Minor nutrients are:
d Sulfur - sulfates
e Potassium
f Trace inorganics - magnesium, iron,
etc.
g Trace organics-vitamins, amino
acids
2 Sources of - See Fig. 1
a Atmosphere
b Groundwater - springs
c Storm water or surface runoff
d Waste waters - domestic sewage and
industrial wastes.
3 Significance of each major nutrient
a Carbon dioxide - See Fig 2
Usually present in great abundance.
Rapidly replenished from atmosphere
and bacterial decomposition of organ-
ic matter. No reasonable possibility
of human control. Nature, however,
does provide some control through
elevated pH levels if carbon dioxide
becomes depleted rapidly.
Nitrogen - Like land plants, certain
algal forms prefer nitrogen in the
form of NH3(NH4+) and others prefer
it in the form of NOs". Both forms
often become depleted during the
growing season and reach maximum
concentrations during the winter
season. A level of 0. 30 mg/1 of
inorganic nitrogen at the time of the
spring turnover is considered to be
the maxmum permissible level .
All natural surface waters are
saturated with nitrogen gas. This
serves as a source of nitrogen for
bacteria and algae capable of fixing
it.
Phosphorus - A key element in all
plant and animal nutrition. The
critical level is considered to be
001 mg/l at the time of the spring
turnover . Phosphorus is needed
to sustain nitrogen fixing forms .
D Practice Of
1 Exclusion of light - Practice well
established in distribution system
reservoirs but impractical on large
storage reservoirs.
2 Nutrient limitation
a Control of surface run-off quality
1) Agricultural
2) Other
b Diversion of sewage plant effluents
1) Madison, Wisconsin
2) Detroit Lakes, Minnesota
3) Pending - State College, Pa.
c Tertiary treatment of sewage
1) Nitrogen removal - Because of
the several forms is very difficult.
V 3-2
-------
ATMOSPHERE
STORM
WATER
(SURFACE
RUN-OFF)
WASTE
WATER
(DOM. SEW.
IND. WASTE
GROUND
WATER
(SPRINGS)
LAKE
OR
RESERVOIR
FIG. I SOURCES OF FERTILIZING
MATERIALS OF CONCERN
IN SURFACE WATERS
I
cc
o
o
£
o
p
3
O
n
o
P
o
"1
tn
-------
CO
£>
ATMOSPHERE
HCO~ + OH
o
o
o
!U
O
C/3
c
V
n
ro
£
p
(D
FIG. 2 CARBON DIOXIDE - BICARBONATE - CARBONATE - HYDROXIDE
RELATIONSHIPS IN NATURAL WATERS
-------
Control of Plankton in Surface Waters
Also, may be unsuccessful in
control unless phosphorus is con-
trolled, too, because of nitrogen
fixing forms.
2) Phosphorus removal - Phosphorus
can be effectively removed by
coagulation methods employing
lime, alum or ferric salts. It
is expensive and no one has
proven its value beyond laboratory
experiments.
d By Biological Engineering
Laboratory studies have shown that
effluents essentially free of plant
fertilizing elements can be produced
by biological treatment of wastes
with proper ratios of C to N and P.
3 Experiences
a Madison
b Detroit Lakes
c State College
d Lake Winnisquam, N. H.
E Practical Aspects
1 Diversion
2 Nutrient control
REFERENCE
Mullican, Hugh F. (CorneU Univ.)
Management of Aquatic'Vascular Plants
and Algae, pp. 464-482. (in Eutro-
phication: Causes, Consequences, and
Correction. Nat. Acad. Sci.) 1969.
This outline was prepared by C. N. Sawyer,
Director of Research, Metcalf & Eddy
Engineers, Boston, Massachusetts.
V 3-5
-------
CONTROL OF INTERFERENCE ORGANISMS IN WATER SUPPLIES
I NECESSITY FOR DATA
A Information on the number, kinds, and
effects of interference organisms in a
particular water supply is essential for
determining adequate control measures.
B Collection of the biological data should be
on a regular routine basis.
C Interpretation of data requires information
on relationship of number and kinds of
organisms to the effects produced.
D It is generally more satisfactory to an-
ticipate and prevent problems due to these
organisms than it is to cope with them later.
II CONTROL IN RAW WATER SUPPLY
A Use of algicides
1 Application of an algicide is to prevent
or destroy excessive growths of algae
which occur as blooms, mats or a high
concentration of plankton.
2 Algicide may be applied to control even
low concentrations of certain algae such
as Synura.
3 Copper sulfate is the only algicide in
common use at present.
a Application may be by dusting,
spraying or dissolving from a porous
container over all or part of the water
surface, or by continuous feeding
of the algicide at the intake of the
reservoir or pre-treatment basin.
b Effective dosage depends upon the
Alkalinity and pH and temperature
of the water and the amount and
kinds of algae to be controlled.
Bartch states that the following
arbitrary dosages have been found
to be generally effective and safe:
M.O. alkalinity > 50 p.p. m. =
2 p. p. m. in the surface foot of
water only (5.4 pounds per acre).
M. O. alkalinity < 50 p. p. m. = 0. 3
p. p. m. in total volume of water
(0. 9 pound per acre foot).
c Application of copper sulfate should
be limited to the minimum effective
dosage because of its corrosive
properties, and its toxicity to fish and
other aquatic animals.
4 Other algicides
a Promising types include inorganic
salts, organic salts, rosin amines,
antibiotics, quinonesj substituted
hydrocarbons, quaternary ammonium
compounds, amide derivatives and
phenols. Cuprichloramine which is
a combination of copper, chlorine and
ammonia, and also chlorine dioxide
have shown promise as general algi-
cides.
b For domestic water supplies they will
have to be not only economically fea-
sible but nontoxic to animal life and
to green plants other than algae.
c Due to higher costs they will prob-
ably be used only when adequate plank-
ton and algal records are kept, which
would permit early localized treat-
ment.
d Algicides selectively toxic to the
particular algae of greatest signifi-
cance would be useful.
5 Mechanical removal or spreading out
to permit rapid drying may be the sim-
plest way of handling massive growths
which are detached and washed ashore.
6 Turbidity due to silt keeps down the
plankton population. In shallow reservoirs,
fish which stir up the bottom mud will
aid in keeping turbidity due to silt high.
7 Provisions for keeping the amounts of
nutrients to a minimum may be em-
phasized more in the future.
8 For new reservoirs, clearing the site
BI. MIC. con. 6b. 4. 70
V 4-1
-------
Control of Interference Organisms in Water Supplies
of vegetation and organic debris before
filling will reduce the algal nutrients.
Steep rather than gentle slopes will
reduce the areas which allow marginal
growths to occur.
Ill CONTROL IN TREATMENT PLANT
A Coagulation and sedimentation
1 When well regulated they often will re-
move 90 per cent or more of the plank-
ton.
2 With low plankton counts, a coagulant
aid may be required.
3 Frequent removal of sludge from the
basins, especially during the warm
seasons may help to reduce tastes and
odors originating from decomposing
organic sediment.
B Sand filtration
1 Both slow and rapid sand filters tend to
reduce the plankton count of the effluent by
90 per cent or more, when well regu-
lated.
2 For rapid filters, accumulated plankton
can be removed or reduced by surface
scraping and by back washing.
C Micro-straining
1 This involves the passing of the water
through a finely woven fabric of stain-
less steel. All but the smaller plankton
organisms tend to be removed from the
water. It is being used in some treat-
ment plants in England and elsewhere.
D Activated carbon
1 The slightly soluble, organic, taste
and odor compounds tend to be readily
adsorbed by the activated carbon. It
is probably most often applied prior to
coagulation, but may be used prior to
filtration or in the raw water.
E Chlorination
1 Treatment with chlorine is practiced
primarily to destroy pathogenic organ-
isms. The dosages commonly used are
toxic also to many algae and to some of
the other groups of aquatic organisms.
However, dead as well as living organ-
isms are often capable of causing tastes
and odors and of clogging filters.
The depth and position of the intake for
entrance of raw water into the treatment
plant may determine the kinds and amount
of plankton which will be drawn into the
plant. Plankton algae generally are more
concentrated near the surface of the water
in lakes and reservoirs.
IV CONTROL IN DISTRIBUTION SYSTEM
A Maintenance of a chlorine residual con-
trols the chlorine sensitive organisms.
B Other pesticides such as cuprichloramine
have been used in attempts to control the
resistant organisms such as worms,
nematodes and copepod eggs.
C Flushing of infested portions of the system,
especially dead ends may be practiced.
D Covering of treated water reservoirs to
prevent the entrance of light will stop the
growth of algae.
E Organisms associated with pipe corrosion
are probably the most active when the water
itself is corrosive.
F Mechanical cleaning of the distribution
system may be an effective but expensive
method of reducing infestations of attached
organisms.
V SUMMARY
A Adequate control is dependent upon ade-
quate procedures for detecting and record-
ing of organisms.
B Control may involve the following-
1 Use of an algicide or pesticide.
V 4-2
-------
Control of Interference Organisms in Water Supplies
2 Mechanical cleaning of distribution
lines, settling basins, and filters,
screens, intake channels and reservoir
margins.
3 Modification of coagulation, filtration,
chemical treatment or location of raw
water intake.
4 Use of adsorbent, such as activated
carbon, for taste and odor substances.
5 Modification of reservoir to reduce the
opportunities for massive growths.
a By covering treated water reservoirs
b By increasing the depth of the water
c By eliminating shallow marginal
areas
d By reducing the amount of fertilizing
nutrients entering the reservoir
e By encouraging a balanced develop-
ment of the aquatic organisms
REFERENCE
Mackenthun, Kenneth M. The Practice of
Water Pollution Biology. FWPCA.
U.S. Dept. of Interior, Washington, DC.
1969.
This outline was prepared by C. M. Palmer,
Former Aquatic Biologist, Biological Treat-
ment Research Activities, Cincinnati Water
Research Laboratory, FWPCA, SEC.
V 4-3
-------
NUTRIENTS: THE BASIS OF PRODUCTIVITY
I INTRODUCTION
A Nutrients of importance include macro-
nutrients: those needed m large quantities,
and micronutnents: those needed in small
amounts.
B These nutrients are important because
they promote biological responses which
may interfere with some desired use of
the water by man.
C Other factors (e. g. temperature, light)
affect the use of these nutrients and should
be considered in an evaluation of the effects
of nutrients upon the ecosystem.
II Algae, bacteria, fungi and aquatic plants
are the forms of life which nutrients affect
most directly.
A Algae are of Several Types
1 Phytoplankton are small algae suspended
in the water and form the basis of pro-
ductivity in the aquatic environment.
2 Benthic algae are those forms anchored
to substrates of rock and bottom
materials.
B Aquatic plants are of several types. In
general they may be referred to as rooted
or floating forms.
C Heterotrophic bacteria are fungi which
respond to organic nutrients introduced
into water. Autotrophic bacteria may re-
spond and grow due to inorganic nutrient
sources.
Ill BIOLOGICAL LAWS
A Liebig's "law" of the minimum: the essen-
tial material available in amounts most
closely approaching the critical minimum
needed will tend to be the limiting.
B Shelford's "law" of tolerance: survival
of an organism can be controlled by the
quantitative or qualitative deficiency or
excess with respect to any one of several
factors which may approach the limits of
tolerance for that organism.
C Q10 "law": with a temperature increase
of 10 degrees centigrade metabolic pro-
cesses (rates) are approximately doubled.
IV The process of photosynthesis is the fixa-
tion of the sun's energy with the production
of organic matter by plants.
A The general reaction is given below:
CO,
B Chlorophyll contains basicaUy C, O, H,
N and Mg, and in general makes up about
5% of the dry weight of algal cells.
V MEASUREMENT OF PHOTOSYNTHESIS
A Oxygen production can be used as a
measure of photosynthesis because for
each mole of CO2 reduced to organic
carbon one mole of free oxygen is liberated.
1 The value of O2/CO2 has been found
experimentally to be 1. 25 rather than
1.0.
B CO2 Assimilation
1 The CO2 taken up by algae does not all
originate from the dissolved gas. Some
algae can use bicarbonate directly as
a source of carbon.
W.RE.ntr.2c.4.70
V 5-1
-------
Nutrients: The Basis of Productivity
2 Hence measurement of CO2 uptake from
water is a complicated problem which
must consider pH, HCO^, and COg"
concentrations.
C Fixation of Carbon-14
1 The use of C14 as a tracer of C12 in
plant metabolism and productivity
estimation has been widely used since
the early nineteen fifties.
2 In this method a known amount of C14
is added to the water and after a period
of time the proportion of C*-4 in the
plant cells to C14 added is found. The
amount of carbon assimilated is then
estimated from the following equation.
activity of
phytoplankton
activity of
added
(K) =
total carbon
assimilated
total carbon
available
3 Where K is a constant relating to the
slower uptake of C14.
4 The total carbon available is determined
chemically.
D Uptake of Mineral Nutrients
1 The measurement of depletion of
nutrients in solution has been tried
but found unreliable.
E Chlorophyll
1 The quantity of chlorophyll present has
been found to bear some relation to
productivity but not a reliable one.
VI Nutrients of significance in the growth and
production of algae and plants are discussed
below.
A Carbon
1 Sources
a Gaseous CO2
b HCOl
c CO-
d Other carbon compounds
2 Effects of the removal of carbon upon
the water
a Lowered pH
b Deposition of CaCO,
3 The quantity of carbon available is
great and it usually is not a limiting
factor.
B Nitrogen
1 Nitrogen can be taken up by most algae
as either ammonium salts or as nitrates.
Nitrites can also be used but a high con-
centration is usually inhibitory. Some
blue green algae can fix atmospheric
nitrogen. Certain algae varieties
require supplementary amines, growth
factors, etc.
2 The quantity of nitrogen in waters has
definitely been shown to limit algal
populations.
C Phosphorus
1 Phosphate seems to be the only inorganic
source of this nutrient.
2 Limiting concentrations of P have been
found to range from . 05 ppm at a mini-
mum and an inhibitory affect if P con-
centrations exceed 20 ppm.
3 Optimum concentrations have been found
to range from .018 ppm to 15 ppm.
4 Storage of inorganic phosphate by algae
has been demonstrated. The extent of
this storage may reach 80% of the total
phosphorus in algal cells.
D Silicon
1 Nutrient ratios in the algal cells of
some areas have been found to be Si 23:
N 16: PI. It can be seen from this
ratio that silicon is an important ele-
ment in algal growth.
V 5-2
-------
Nutrients: The Basis of Productivity
2 Silicon is especially important in the VII
population growth of diatoms and may
be the limiting growth factor in these
populations. A
E Inorganic micronutrients - Many elements
are needed in very small quantities by
algal cells. Some of these have a known
function in algal metabolism; others do
not.
1 Mg is a cation of major importance in
the chlorophyll molecule.
2 Co is known to be necessary for vitamin
B12-
3 Mn is necessary for several enzyme
systems.
4 Mo, V, Zn, and Cu are necessary but
these functions are not as well known.
F Organic Micronutrients
1 Of 179 algal strains investigated about
40% required vitamin supplementation
for optimum growth. Principal growth
factors that were not synthesized in
sufficient quantity are given as follows
along with the percentage of the vitamin B
deficient strains showing marked pro-
ductivity gain after supplementation:
addition increased growth on
of the strains.
b Thiamin addition increased growth
on 53% of the strains.
c Biotm addition increased growth on
10% of the strains.
Algae can use and may require many
organic compounds depending upon
environmental conditions and the ability
of the organism to synthesize required
building blocks from mineral forms of
C.N. & P. This is an area for con-
tinued investigation with many unappre-
ciated or vaguely understood ecological
factors.
PROBLEMS AND BENEFITS RESULTING
FROM ALGAE PRODUCTION
Problems to man may result when the
total "primary production" by algae leads
to an increase in the total organic content
of the water that interferes with a desired
use.
1 This may consist of a high algal popu-
lation that produces a water with high
turbidity, taste and odor, or other
undesirable effect. High respiratory
needs may lead to nocturnal oxygen
deficit.
2 Certain algae may cause tastes and
odors, clog filters; or otherwise inter-
fere with potable water processing.
3 Death of large algal populations may
lead to tastes and/or odors through
bacterial decomposition. Oxygen
deficits may result at any time of day
in this process. Deposition of masses
of organic sediment or sludge may be
considerable.
4 Other problems might be cited.
The primary production of algae can also
serve as a supply of food to consumer
organisms (animals), resulting in increased
production at several (trophic) levels:
zodmicrobes. microinvertebrates,
macroinvertebrates, fishes.
Earlier notation cited the release of
oxygen during utilization of CO2 during
algal photosynthesis. This encourages
fungal or bacterial breakdown of
pollutants.
Photosynthesis occurs in the presence
of adequate light and favorable condi-
tions. In darkness, the cells continue
to respire and may consume more
oxygen than they produced because
photosynthesis increases the organic
load.
V 5-3
-------
Nutrients: The Basis of Productivity
Photosynthesis tends to occur at the
surface where light intensity is greatest.
Poor vertical mixing would result in
stratification of water supersaturated
with oxygen over oxygen deficient water.
Depending upon conditions, a significant
fraction of the oxygen could be lost to
the atmosphere.
Increased productivity may result in
temporary reduction of the free dissolved
nutrient level in the water but harvesting
at some level is essential to prevent
later recycle.
VIE CYCLE OF NUTRIENTS
A Once nutrients enter a body of water they
are cycled through a food chain.
B Factors affecting this food chain (e.g.
toxicity, removal) will affect the con-
centration and distribution of the nutrients.
2 Lewin, Ralph A. Physiology and
Biochemistry of Algae. Academic
Press. 1962.
3 Odum, Eugene P. Fundamentals of
Ecology. W.B. Saunders Co. 1959.
4 Odum, H. T. Primary Production in
Flowing Waters. Limnology and
Oceanography. 1(2):102-117.
April 1956.
5 Ryther, John H. The Measurement of
Primary Production. Limnology and
Oceanography. l(2):72-84. April 1956.
6 Verduin, Jacob. Primary Production in
Lakes. Limnology and Oceanography.
1(2):85-91. April 1956.
ACKNOWLEDGEMENT:
This outline contains certain material
submitted by F. J. Ludzack and H.W. Jackson.
REFERENCES
1 Golterman, H.L. and Clymo, R.S.
Chemical Environment in the Aquatic
Habitat (Proc. of an IBP - symposium,
Amsterdam and Nieuwersluis Oct. 1966).
322pp. (N.V. Noord-Hollandsche
Uitgevers Maatschappij, Amsterdam.
8.95).
This outline was prepared by Michael E.
Bender, Biologist, Formerly with Training
Activities, Ohio Basin Region, SEC.
V 5-4
-------
CHAPTER VI
RELATED STUDIES
The Problem of Synthetic Organic Wastes
Beneficial Aspects of Algae
Behavior of Radionuclides in Food Chains -
Freshwater Studies
FWPCA Responsibilities for Water Quality Standards
Marine and Estuarine Plankton
Attached Growths (Periphyton or Aufwuchs)
Artificial and Related Substances - References
2
3
4
5
6
7
8
-------
THE PROBLEM OF SYNTHETIC ORGANIC WASTES
I Sources of organic chemicals in water
are varied and of differing complexity. * ^
A Natural pollutants, such as algae, actino-
mycetes, etc. contribute to organic
pollution.
1 Tastes and odors associated with these
materials are probably not merely a
result of decomposition, but are closely
associated with materials produced
during the life cycle of the organisms
and plants.
2 Discharge of nutrients in the form of
phosphorus and nitrogen compounds
from domestic or other wastes fre-
quently stimulate the production of
natural pollutants.
B Industrial wastes, due to the rate of
population increase and industrial ex-
pansion, have made the problem of
effective water treatment an acute one in
many places.
1 The production of synthetic organic
chemicals has risen steadily over the
past years, representing many new
and complex products - and of im-
portance to us - new and complex wastes.
2 The ideal method of handling industrial
waste is at its source.
a However, what is often considered
good treatment, still results in
materials present in sufficient
quantities to affect the taste and odor
of water.
b Many problems are caused by slug
discharges, often accidental.
C Domestic wastes in various stages of
treatment.
D Miscellaneous sources also contribute to
the problem.
1 Wastes from private and commercial
boats.
2 Chemicals applied to the land may be
washed into streams.
3 Chemicals applied directly to water.
a Evaporation control
b Killing off rough fish
c Aquatic plant control
II Concentrations of organic chemicals
in water, even in comparatively minor
quantities may cause difficulties
A Wastes may contain from a few mg/1 to
several hundred mg/1 of organic
contaminants.
B Surface waters may contain from a few
jjig/1 of organics to several mg/1.
1 Some of the chemicals isolated from
water, along with the concentrations
which can be detected by odor, are:'^'
Concentration
Substance
Detectable*,
Formalde hyde
Picolmes
Phenohcs
Xylenes
Refinery hydrocarbons
Petrochemical waste
Phenyl ether
Chlorinated phenohcs
50,
500
250
300
25
15
13
1
000
- 1,000
- 4, 000
- 1, 000
- 50
- 100
- 100
^Concentrations were determined by taking
the median of 4-12 observations.
CH.OTS. 40a. 4.70
VI 2-1
-------
The Problem of Synthetic Organic Wastes
III The damaging effects of organics in water
are becoming more apparent.
A Taste and odor in water is usually the first
noticed effect from organics. This is a
serious public relations and economic
problem; it also may be a health problem.
B Organic contaminants may interfere with
coagulation, damage ion exchangers, and
create chlorine and carbon demand.
C In the stream they may have adverse
effects on aquatic forms that support higher
aquatic life, cause off-flavors in fish flesh,
or have direct toxic effects on fish.
IV The methods of study employed in the
collection and identification of organic
chemicals in water involve physical and
chemical methods and instrumental analysis.
The comparatively small amounts of
organic materials may be concentrated
by adsorption on activated carbon.
1 This carbon is then extracted with
appropriate organic solvents, the
solvent extract is taken to dry ness, the
weighed extract is subjected to solubility
group separation, and these individual
groups may then be analyzed by various
methods.
2 Employing the above method on Ohio
River water, the following results were
obtained:*3^
Chemical Group
Water solubles
Ether and water
insolubles
Neutral
Amine
Weak acid
Strong acid
Amphoteric
Loss
%of
Total
20
22
14
4
8
6
10
16
TOC
in (ig/1
860
110
3
575
645
365
5, 000+
--
Relative Odor
Contribution
23
200
4,670
7
12
16
--
B Chemical separation and analyses may be
accomplished by means of column chromato-
graphy, formation by derivatives, gas
chromatography, infrared and ultraviolet
spectroscopy, x-ray diffraction, etc.
1 Specific organic chemicals recovered
from river and drinking waters by
these methods include: synthetic
detergents (ABS), phenylether, phenol,
DDT, aidrin, o-nitrochlorobenzene,
a-conedendrin, and xylene.
V Some of the types of problems that may
be attributed to organic wastes, and more
specifically to problems of taste and odor,
may be represented by the following examples:
VI 2-2
-------
The Problem of Synthetic _ Organic Wastes
A By applying the previously mentioned
carbon adsorption method, the odor po-
tential of organic pollutants and the
dilution necessary to reduce this odor
potential to a barely perceptible level has
been determined:'^'
Industry Source
Brewery
Chemical
Corn Refining
Meat Packing
Metal Fabrication
Paint
Pharmaceutical
Refinery
Soap
Cone. Required for
Detectable Odor ng/1
CHC13
Sol. Org.
770
28
1,000
1,200
890
390
290
84
900
Total
Org.
1,400
32
3,600
3,600
1,600
1,000
340
510
1,800
Dilution Factor
CHC13
Sol. Org.
14
11,000
1.4
92
2.8
69
10
780
640
Total
Org.
86
14, 000
2. 1
140
4.8
98
32
760
350
Of all the organic pollutants that can affect
the taste and odor of drinking water,
phenol has been the most extensively
studied.
1 The potential sources of this chemical,
both natural and synthetic, have been
discussed. (5)
2 The course of chlorination of phenol,
a common method of treatment in the
water plant, has been shown to proceed
by a process which starts with the pure
compound (in itself relatively tasteless)
and proceeds through strong-tasting
intermediates to tasteless end
products. (6)
1970 the petrochemical production on a
tonnage basis may be equal to 41% of all
chemicals.
1 The three principal groups of petro-
chemicals are the paraffins, the
naphthenes, and the aromatics. From
these, over 200 basic products are
manufactured, having thousands of
subordinate uses.
2 Correspondingly, more than 100 identi-
fiable compounds have been found in
waste streams from petrochemical
processes.
BIBLIOGRAPHY
C The effects of petrochemical wastes^
on water quality are becoming increasingly
important, it has been predicted that by
1 Middleton, F. M. Taste and Odor Sources
and Methods of Measurement. Taste
and Odor Control Journal. 26:1. 1960.
VI 2-3
-------
The Problem of Synthetic Organic Wastes
2 Middleton, F.M., Rosen, A.A., and
Burttschell, R. H. Taste and Odor
Research Tools for Water Utilities.
Jour. A.W. W.A. 50:21. 1958.
3 Anonymous. Objectionable Organic Con-
taminants in Water. San. Eng. Center
Activ.Rep. No. 25, 1955.
4 Sproul, O. J., and Ryckman, D. W. The
Significance of Trace Organics in Water
Pollution. PCF 33; 1188. 1961.
5 Hoak, R. D. The Causes of Tastes and
Odors in Drinking Water. Water and
Sewage Works. ^04:243. 1957.
6 Burttschell, R. H., Rosen, A. A.,
Middleton, F.M., and Ettinger, M. B.
Chlorine Derivatives of Phenol Caus-
ing Taste and Odor. Jour. A. W. W. A.
5T205. 1959.
7 Gloyna, E.F., and Malina, Jr., J. F.
Petrochemical Wastes Effects on
Water. Indus. Water & Wastes, 7_:
#5. 134. Sept.-Oct. 1962.
8 Baker, R. A. Problems of Tastes and
Odors. WPCF. 33:1099. 1961.
This outline was prepared by R. L. Booth,
Chief, Analyses Unit, Analytical Quality
Control Laboratory, 1014 Broadway,
Cincinnati, OH 45202.
VI 2-4
-------
BEHAVIOR OF RADIONUCLIDES IN FOOD CHAINS
FRESH WATER STUDIES
INTRODUCTION
Fresh water lakes and streams, in their
traditional roll of receiving and transporting
all manner of materials, seem destined to
continue this function in the atomic age and
have radionuclides. In many cases the water
may serve as an effective medium through
which the nuclides can become incorporated
into living organisms. Ultimately some of the
radioactive materials may be deposited in fish
or crops which are eaten by man and thus
provide an additional source of radiation
exposure.
This discussion is primarily directed toward
the mechanisms through which aquatic forms
may accumulate radioactive materials from
their environment and the manner in which we
may expect certain nuclides to become
distributed in biological systems of fresh
water lakes and streams. In a broad sense,
the subject of the behavior of radionuclides in
fresh water food chains might also include
irrigated crops, and animals which drink from
streams and ponds. Such problems, are how-
ever, more closely allied with terrestial
studies and in this case consideration will be
given only to those forms which actually live
in the water.
SOURCES
Fresh water streams may receive radioactive
materials from a great variety of sources.
Naturally occurring radioisotopes removed
from the atmosphere and eroded from radio-
active ores have been carried in both surface
and underground waters since early in the
history^ the earth. Typically, U238. Ra ,
and Th are present in measurable amounts,
but other isotopes such as K and C are
also present. The background levels are in
the range of 10 ...to 10 |ic/ml for most
surface waters but values in excess of
10 uc/ml have been reported for certain hot
springs of high radium content. In most
streams the natural concentration of the heavy
metals is so low and their uptake by aquatic
forms is so small that they contribute but
little to the radioactive content of the organisms.
Most of the natural radioactivity in the biota
originates from K which, in fish, amounts
to about 3 X10'6 jic/g.
With the creation of a demand for large amounts
of uranium as a basic source of nuclear energy
the uranium ore refining industry has expanded
rapidly during the past decade. As pointed
out by Tsivoglou, et al. ' , the concentrations
of radioelements of the uranium-radium
family are substantially increased in streams
which receive the effluent directly from
uranium mills or seepage from ponds used to
retain their tailings. The concentration of
radium in the biota of the streams is likewise
increased.
Liquid wastes from the fabrication of refined
uranium into fuel for reactors can be another
source of naturally radioactive isotopes to
streams. In comparison with releases at the
mills and with the problems which accompany
irradiation of the fuel, however, the contribu-
tion from the fabrication plants will usually
be quite insignificant.
The large quantities of artifically radioactive
nuclides produced in reactors form a new type
of disposal problem, the handling of which
requires a technology considerably more com-
plex than that associated with the natural
isotopes. Dependent upon individual design,
the reactors themselves may or may not
contribute significant quantities of radio-
nuclides to fresh water streams. The mega-
curie quantities of fission products which
accumulate in the reactor fuel do not, under
ordinary circumstances, escape from the fuel
while itis in the reactor. As pointed out by
Terrill , "Present practice involves extensive
control measures through design and operation
to confine the fission products within the
reactor core and its immediate environs. "
Failure of the cladding of fuel elements does,
of course, provide the opportunity for entry
of fission products into the primary coolant
stream or moderator. Under normal circum-
stances, the principal source of radioactive
materials in the fluids which pass through the
reactor core is the activation of materials
which are exposed to the neutron flux. These
activation products may originate from materials
dissolved in the coolant or they may corrode from
11. l.C. (6.62)
VI 4-1
-------
Behavior of Radionuelides' in Food Chains - Fresh Water Studies
the surfaces of the fuel elements, cooling
system, or structural materials. In the Han-
ford-type reactor, the coolant is filtered
Columbia River water which is returned to
the river after a single pass through the re-
actor. Figure 1 shows the isotopes which
predominate in effluent from the Hanford re-
actors four hours after irradiation. Many
of these have such short half-lives that they
have decayed to very low levels by the end
of the first day. Minor amounts of fission
products are also found in the Hanford reac-
tor effluent not only as a consequence of the
occasional failure of a fuel element, but also
because of natural amounts of fissionable
uranium in the Columbia River water. The
release of radioactive effluent to the Colum-
bia River is accompanied by extensive monit-
oring, both before and after discharge, in
order to provide assurance that the radiation
exposures to people who subsequently use
the river or eat fish or irrigated crops are
well within permissible limits.
Figure 1
-
Mn =
}
1
Cu«<
«,*<
Q COMPOSITION AT 4 HOURS
READ SCALE AT LEFT
rrrq COMPOSITION AT 24 HOURS
I'.:'.':* READ SCALE AT RIGHT
nRan^ra:
C,5I Np233 A,7« 5iJI 2n«> Q,Ti
Sr92
a a
RADIO-ELEMENT CONTENT OF REACTOR
EFFLUENT 4 HOURS AND 24 HOURS AFTER
IRRADIATION. (Radioelements marked * ore
are routinely measured).
"Radioactive Waste Management Operations
at the Hanford Works", included in Con-
gressional Hearings on Industrial Radio-
active Waste Disposal (1959).
Thd kinds and amounts of radionuclides dis-
charged to the Columbia River with the Han-
ford reactor effluent should not be considered
as characteristic of other types of reactors.
The concentrations of various activation
products which build up in the primary cir-
culating fluid will be dependent upon the in-
tended chemical composition of the fluid,
impurities present, the corrosion of cladding
on fuel elements, piping and vessels, film
formation, neutron energy and flux, radioactive
half-life, and the effectiveness of ion-exchange
"clean-up" loops where these are used. Sig-
nificant amounts of activation products of
common elements ranging from tritium to
neptunium can conceivably be present. In
most cases there probably will not be inten-
tional release of radioactive materials accumu-
lated in the re-circulated fluids directly to fresh
water streams. However, minor leakage from
the complex piping and fuel handling systems
into the plant effluents appears almost inevit-
able.
Excluding nuclear detonations, the largest
potential source of radionuclides to the environ-
ment is the fission products which accumulate
in reactor fuels. At the present time, the
bulk of the high-level waste which results from
the processing of the fuel elements, is confined
in some artificial or natural container. Even
so, large quantities of water with low or inter-
mediate concentrations of radionuclides result
from the later stages of most separation pro-
cesses and these must usually be disposed of
at, or very near, the processing plant. In
many cases, disposal of such wastes will be
to the ground or to pits on the surface in order
to take advantage of the ion-exchange capacity
of the soil and the time lags which allow for
decay of the isotopes with short and intermedi-
ate half-lives. At the Hanford plant the cumu-
lative volume of intermediate-level waste dis-
charged to the ground through 1958 amounted
to nearly 4 billion gallons, containing nearly
2 million curies of beta-emitters. The topo-
graphy, soil structure, and geology at Han-
ford favor retention and none of this material
has been detected in the Columbia River^4^
Conditions for surface disposal are less favor-
able at other existing major installations,
and wastes containing fission products dis-
charged to pits at both Oak Ridge<5) and Chalk
River*6' have seeped through the soil and
appeared in streams and lakes which receive
the drainage.
Radioactive wastes may also enter fresh water
streams via the drains from industrial, medical.
VI 4-2
-------
Behavior of Radionuclides in Food Chains - F^-esh Water Studies
or research facilities which use isotopes.
Although the amounts involved are compara-
tively small, such sources should not be
entirely disregarded since the Oak Ridge
National Laboratory is now shipping on the
order of 150, 000 curies of isotopes a year
to domestic users.
On the basis of high yield from nuclear fission,
significantly long half-life, and biological
importance, the fission products which one
would expect to find in greatest abundance
in the biota of fresh water streams and lakes
are: Sr89, Sr9<>, Y91, Zr95+ Nb95, Ru106,
1*31, cs137, Ba140. Ce144, and one or more
rare earths. In addition to these, neutron
activation products which appear to be of
importance include: P32, Sc46, Cr51, Mn54,
Fe55, Fe59. Co", Co60 and Zn". In the
immediate proximity to reactors, .very short-
lived isotopes such as Na , Cu^4, and As7**
may be detected.
As a result of the use of nuclear energy, the
associated mining and refinement of fission-
able materials, and the widespread use of
radioisotopes, the radioactivity in a few sur-
face waters of the world has now been in-
creased above the original background level.
As the use of atomic energy and its byproducts
becomes more extensive, the numbers of
rivers and lakes which receive radionuclides
will increase, and the organisms which in-
habit these waters will also acquire the radio-
active materials. Some appreciation of the
mechanisms involved in the uptake of radio-
nuclides by aquatic organisms is necessary
to anticipate potential sources of exposure
to man, and, if necessary, to effectively
control or reduce the amounts accumulated.
UPTAKE BY LIVING ORGANISMS
Radioactive materials present in the water
may be taken up by aquatic organisms via
three different mechanisms: 1. adsorption
onto exposed surface areas, 2. absorption
through membranes exposed to the surround-
ing water, possibly with the aid of active
transport mechanisms, and 3. engulfment
of food or inert particles which contain the
nuclides. The relative importance of these
mechanisms is different for various kinds of
organisms and even similar species. It also
fluctuates widely between different elements
and different environments.
Adsorption, which is a physical process,
occurs very rapidly. It is of greatest signifi-
cance among those organisms which nave
large surface to volume ratios such as sponge
and diatoms. A clear distinction between
adsorption and biological absorption is not
easy to make however, since the physical
process of adsorption may be a necessary
primary step in the movement of ions into
the cell. Films of bacteria and other micro-
organisms may also fix radioactive materials
on the surfaces of organisms to give an ad-
sorption-like effect.
The process of absorption is obviously of
prime importance to the plants since this is
the means by which nutrient materials are
removed from solution and metabolized.
Many non-essential elements will also enter
the plant cells because of passive diffusion
through the membranes or because the or-
ganism is handling them in a manner similar
to the essential elements. The uptake of radio-
active materials by the plants is, of paramount
importance to the higher forms since the
plants form the food base.
Direct absorption of some radioisotopes from
the water also occurs in aquatic animals, in-
cluding the fish, with the gill membranes
probably playing the important role. Such
direct uptake by fresh water fish has been
demonstrated for Sr90(7) (8) (9), Ba140 +
Lal40 (7), Ca45 (10) (11) (12), P32 (13) Na2*
(7), and Csl37 (14). However, the dominant
means by which many radioactive materials
are accumulated by aquatic animals is through
ingestion, since this is the manner in which
they obtain the bulk of their nutrient materials.
Ingestion does not insure that the isotope will
be assimilated through the gut. Some mater-
ials may pass through the gut with virtually
no uptake, and seldom, if ever, will all of
the ingested radioactive material be metaboli-
zed. For example, Watson*15* found that after
24 hours, rainbow trout retained about 60 per
cent of the radiophosphorus which they ingested.
Schiffman*16) found that only about 7 per cent
of Sr90 incorporated into natural food organ-
isms was retained by trout after one day.
VI 4-3
-------
Behavior of Radionuclides in Food Chains - Fresh Water Studies
CONCENTRATION FACTORS
The uptake of radionuclides by organisms is
obviously not a phenomenon which is peculiar
to the radioactive forms of the elements.
Rather, the radioisotope acts only as a tracer
to demonstrate the difference between the
abundance of the element in the water and in
the organism. The so-called "concentration
factor"
of organism)
((AC /ml of water )
for any radionuclide cannot exceed the ratio
which exists between the concentrations of
the element in the organism and in the water.
On this basis it should be possible to predict
the ultimate concentration factors for various
radionuclides from conventional quantitative
chemical measurements, provided the nuclide
has become distributed throughout the system
and is uniformly mixed with its stable counter-
part. In practice, however, it may be quite
difficult to measure the concentration of some
elements because of the minute quantities
involved.
Estimation of concentration factors for fresh
water organisms involves, at the present
time, a considerable amount of speculation.
While the concentration of some elements in
a variety of marine organisms is available; (17)
and the chemical composition of sea water is
relatively stable, there is very little infor-
mation readily available on the chemical com-
position of fresh water organisms. Chemistry
of various lakes and streams differs so great-
ly from one region to another that no general-
izations can be made. The picture is clouded
further by the fact that the concentration of
many elements in aquatic forms is partially
dependent upon their abundance in the surround'
ing water. Although it is virtually impossible
to write down probable concentration factors
which will apply to more than one locality,
an indication of the differences in the orders
of magnitude of the concentration for a few
of the common elements in some organisms
and in water is shown in Table 1.
It is evident then, that radioisotopes of the
different elements will be concentrated to
various amounts by the different organisms.
The concentration 'of phosphorus in Columbia
River water is about 0. 01 ppm, while in the
fish it amount* to about 6, 000 ppm. These
values would lead to an estimated concentra-
tion factor of about 600, 000 for the fish.
Since radiophosphorus is one of the isotopes
present in the effluent which is discharged in-
to the Columbia River from the Hanford reac-
tors, the actual concentration factor for this
nuclide can be measured. In contrast to the
theoretical value of 600, 000, the maximum
observed value is only about 100, 000. The
observed value is less because the biological
processes required to deposit and exchange
the phosphorus in the body of the fish are
long in comparison to the radioactive half-
life of P32, which is about 14 days. Conse-
quently, the radioisotope is not uniformly
mixed with all of the stable phosphorus in-
volved in the system. Such a reduction would
not be expected for long-lived isotopes.
Table 1
Concentration (.ug/g wet weight or ml) of
some elements in fresh water organisms and
in some major rivers of the United States
ailloan
SoiJam
Fboqbm,
bleha
Ira.
ttratlB
U(M
(Smrocrm)
1.900
1.900
290
1.900
4. SCO
2
o^o. V
IhMet Urw
(Cidtfli fly)
20
TOO
WO
300
300
0.2
fl*
(nm»i)
10
1000
iooo
3000
1
OO
ViurJ/
In
1
1
40.001
2
<0.01
CUJH,
HUEL
29
200
1.9
200
6J
l.»
l_l Values are only estimates of orders of
magnitude obtained by spectrophotometric
analysis at the Hanford Laboratories. They
are recorded here to illustrate differences
which can exist and are not intended for
use in precision work.
2j Abstracted from Moyle, J. B., "Relation-
ships between the chemistry of Minnesota
Surface Waters and Wildlife Management".
Journal of Wildlife Management 00: 303-320
1956, and Clarke. F. W.. "The Composi-
tion of the River and Lake Waters of the
United States". U. S. Geological Survey,
Prof. Paper 135, 1924. Strontium values
from Alexander, Nusbaum, and MacDonald,
"Strontium and Calcium in Municipal Water
Supplies", A.W.W.A.. Journal 46(7): 643-654.
(1954).
VI 4-4
-------
Behavior of Radionuclides in Food Chains - Fresh Water Studies
From measured amounts of radionuclides in
the effluent from Hanford reactors and in or-
ganisms living in the Columbia River im-
mediately below the reactors, concentration
factors have been calculated for the more a-
bundnat isotopes. These are listed in Table
2 for the season when maximum values occur.
7fi Afi
The concentration factors for As , Sc ,
Cr51, and Cu6^ are quite low in fish in com-
parison to those in algae and insect larvae.
This illustrates that food chains, which on
the one hand provide the mechanism for trans-
ferring radioactive materials from one or-
ganism to another, can also serve,, in some
cases, to reduce the concentration of radio-
nuclides in large animals. Two different
processes can be involved. Where possible
the organisms will eliminate non-essential
elements, and thus there will be a repeated
selection against such nuclides along the food
chain; and short-lived isotopes will decay to
low levels as they pass through the chain.
Table 2*
Observed Concentration Factors
For Significant Isotopes Found In Columbia River Organisms
Isotope
P32
Zn85
cs,37(d)
Sr80
Na24
As76
So46
Cr51
Cu64
AUae(a>
100.000-1.000,000
100. 000
1,000-5.000
10,000
100
10. 000
100, 000
100-1,000
10, 000
Insect'1"
Larvae
100, 000
10,000
1,000
100
100
1,000
1,000
100-1,000
1,000
Fish
100. 000
1,000-10,000
5,000-10,000
1, 000
100-1,000
100
10
10
10
Many of these values are not yet well defined a'nd are subject to revision.
Richardsonium ap.
Sligeoclonliim ap. Hydropayche ap
Data for Cs obtained from a pond environment rather than the Columbia
River and species are different from those listed above.
(Values are based partly on data reported by J. D. Davis, R. W. Perkins,
R.F. Palmer, W.C. Hanson, andJ.F. Cline (18).and R. C. Pendleton and
W.C. Hanson (19).)
This table reproduced from "The Need for Biological Monitoring of Radioactive
Waste Streams, " by R. F. Footer presented at the Nuclear Engineering and
Science Conference April 6-9, 1959, Cleveland, Ohio.
The isotopes which are most likely to have
high concentration factors in fish and other
large aquatic animals are those which are
readily assimilated, which are retained for
relatively long periods of time in various
body structures, and which have long radio-
active half-lives. These are characteristics
which result in relatively high body burdens
for humans as well, and which influence a
low maximum permissible concentration for
the isotope, in drinking water or food.
DIFFERENCES BETWEEN SPECIES
Differences in the concentration factors for
various isotopes among different species leads,
to different levels of radioactivity in the or-
ganisms. Columbia River organisms provide
a good example of this as shown in Figure 2.
The short-lived isotopes contribute significant-
ly to the activity density (quantity of radio-
active materials per unit mass of substance)
of plankton, algae, and sponge. But in the
higher animal forms, about 95 per cent of the
activity originates from P".
Figure 2
Radioactivity in Different Columbia
River Organisms
RELATIVE CONCENTRATION OF RADIOACTIVE MATERIALS
0 Z5 SO 75 100
SESSILE ALGAE
STIGEOCLONIUM
SPONGE
SPONGILLA
CAODIS LARVAE
HYOROPSYCHE
MAY FLY NYMPHS
PARALEPTOPHLEBIA
SNAIL FLESH
STAGNICOLA
SUCKERS
CATOSTOMUS
SHINERS I
RICHARDSONIUS
i
CRAYFISH
ASTACUS
A
P WATER
Davis and Foster, "Bioaccumulation of
Radioisotopes through Aquatic Food
Chains", Ecology 39: 530-535 U05B)
VI--4-5
-------
Behavior of Radionuclides in Food Chains - Fresh Water Studies
32
Because of the dominance of the P , it is of
interest to consider this isotope separately
from the others. Figure 3 shows the relative
concentrations of P in most of the species
included in Figure 2. The different concen-
trations result from a combination of several
factors: the chemical composition of the
various species is different, the moisture
and organic content varies,, and there are
different lag times along the food chain which
permit radioactive decay.
Figure 3
RELATIVE CONCENTRATION OF P32 IN
COLUMBIA RIVER ORGANISMS
As mentioned before, the reduction in the
concentration of a radioisotope of an essen-
tial element along the food chain may occur with
relatively short-lived isotopes because the
ratio of radioactive to non-radioactive atoms
of the element does not remain constant
throughout the entire biological system. This
ratio may properly be called the specific
activity since it is the concentration of the
Isotope per .gram of stable element - in this
case - jie P^2 per.gram of P^l. The Colum-
bia River below the Hanford reactors pro-
vides an unusual opportunity for the study
of the specific activity of P82 in various or-
ganisms of the food chain, since a near
steady state is maintained^ A "fresh" supply
of the isotope is added to the river water at
a more or less constant rate with the reactor
effluent. Since the concentration of stable
phosphorus in the river water is also rela-
tively constant, the specific activity of P32,
remains about the same in the water through-
out much of the year. If there were a near
instantaneous and complete exchange of phos-
phorus between the river water and all aquatic
organisms, then the specific activity of the
organisms would be identical with that of the
water. Figure 4 shows that this is not the
case. For plankton and sessile algae the
specific activity is virtually the same as in
the water: This indicates a very rapid exchange
of phosphorus between these forms and the
water. The specific activity is progressively
less in the insect larvae, snails, fish, and
crayfish (scavengers), indicating that re-
placement of phosphorus in these animals goes
on at a relatively slow rate, and thus the
phosphorus "ages" in the organism.
Figure 4
SPECIFIC ACTIVITY OF COLUMBIA RIVER ORGANISMS
i CRAYFISH
MOIC: THOC WLUCS WOK ESTOMTtD HUt HATtMAL COLLKHD AT I
TIMES MK> AM SUSJCCT TO REVISION
If one compares the specific activity of caddis
fly larvae with that of their food," which is
predominantly plankton for the species rep-
resented here, it can be shown that the aver-
age phosphorus atom remains in the caddis
larvae for about 9 days.
32
The low specific activities of P in the lar-
ger animals results not only from slow phos-
phorus exchange within the animals, but also
from the reduced specific activity of its food.
The effect is thus cumulative along the food
chain. For isotopes which are shorter lived
than P32, the reduction in specific activity
along the food chain may be even more spec-
tacular; for isotopes with relatively long half-
lives the reduction may not be apparent.
It must be remembered, however, that a re-
duction in specific activity along the food chain
results from a combination of two factors:
one is a comparatively short half-life, and
the other is a relatively long retention of the
element in the organisms.
yi 4-6
-------
Behavior of Radionuclides in Food Chains - Fresh Water Studies
EXCHANGE BETWEEN ORGANISMS
Although we talk in terms of food chains with
the implication that radionuclides and other
materials are transferred sequentially from
small food species to larger predatory forms,
such simple, straight-forward systems are
not strictly characteristic of aquatic com-
munities. Such communities might better
be thought of as complex chemical exchange
systems in which there is a kind of equilibrium
established between the concentration of the
nuclide in the water and in the various bio-
logical forms, silt, and other exposed sur-
faces. Several experiments have been carried
out in both the laboratory and the field with
the use of P^ in order to measure the rates
of exchange between various components of
the systems (20-31). Figure 5 is a highly
schematic representation of the kind of phos-
phorus exchange which operates in a fresh
water community. The size of the arrows is
intended to give some conception of the quan-
tity of phosphorus (or radiophosphorus in this
case) moving in a given direction in some
short unit of time. A large fraction of the
available isotope moves rapidly back and forth
between the water and the photosynthetic
plants. Animals which eat the plants obtain
an appreciable part of their radiophosphorus
from the plants, but they also carry on at
least some exchange directly with the water.
A similar arrangement exists at the higher
trophic levels. Each of the groups of organ-
isms contributes some isotope to the sedi-
ments, either through death or via fecal
pellets, with bacteria playing an important
role in decomposition. Some of the isotope
remains with the sediments and, in time, is
buried deeper. However, much of it is re-
leased back into the water and recycled
through the biota.
Figure 5
In many fresh water communities the great-
est mass of material will be made up of the
photosynthetic plants - the primary producers
which constitute the first trophic level. The
mass of successive trophic levels will usually
be smaller and smaller, so that relatively
few carnivorus fish are supported at the top
of the biomass pyramid. However, in some
open-water communities where the primary
trophic level is made up almost exclusively of
phytoplankton, with a rapid "turnover", a
part of the pyramid may be inverted, (32).
Of the total inventory of radiophosphorus and
most other radionuclides, one would expect
to find the greatest fraction held in those or-
ganisms which make up the greatest portion
of the biomass. In fresh water systems this
will usually be the green plants, which also
have rapid exchange rates and an affinity for
a large variety of nuclides. Nevertheless,
only a small fraction may be held in the large
VI 4-7
-------
Behavior of Radionuclides in Food Chains - Fresh Water Studies
fish, this may well be the part of greatest
interest to the radiation protection specialists.
It should be evident then, that radioisotopes
of biologically essential elements, when in-
troduced into streams or lakes, are not apt to
remain exclusively in the water. Rather,
they will rapidly become distributed between
the water, the biomass, and the sediments:
In many cases the portion which is removed
from the water will exceed that which remains.
If it is desired to reduce the specific activity
of a contaminating radionuclide by adding
carrier to the water, consideration must be
given to the probability that there is a greater
reservoir of essential elements in the solids
(including the biota) than is dissolved in fresh
water.
SEASONAL VARIATIONS
Where continued or repetitive release of
radioactive waste occurs, a true equilibrium
in distribution of radionuclides between water
and organisms will seldom be attained. Even
under stable conditions of flow and effluent
discharge, seasonal fluctuations in the composi-
tion and metabolism of the community will keep
the system in a dynamic state. New blooms
of plankton or vascular plants will create a
demand which will draw material from the
water and from other parts of the biomass.
Conversely, the dying-off of large masses of
vegetation may release significant quantities
of radionuclides back into the water where they
will be available to other components of the
community.
Most aquatic animals are cold blooded, and
thus their metabolic rates change with varia-
tions in temperature and so, with the seasons.
Concentrations of those radionuclides affected
by metabolic rates in aquatic organisms will
also fluctuate with the seasons. During the
cold-water period food intake is drastically
reduced. If inges.tion is the principal mode
of uptake of a radionuclide, the supply to the
animal will likewise be reduced, and the
quantity of nuclide in the animal will diminish
as a result of radioactive decay and biological
elimination. The reverse is true when tem-
peratures are high.
Figure 6 shows a smoothed curve for the
activity density of plankton and small fish of
the Columbia River over the period of a year.
Fluctuations in the plankton (predominantly
diatoms), are quite similar to those in the
water since there is a rapid exchange of nu-
clides between the water and the plant cells
by direct absorption and adsorption; tempera-
ture has little effect in this case.
A pronounced decrease in the activity density
of the plankton occurs during the late spring,
because the flow of the Columbia River increases
greatly at this time to provide additional dilu-
tion of the effluent from the reactors. Fluc-
tuations in the activity density of the fish are
on the other hand, more closely related to
temperature. As indicated above, the marked
increase in activity density of the fish during
the time of high temperature reflects an increase
in metabolic rate all along the food chain.
Not only are the fish eating more food, but each
food organism has become more radioactive,
and the effective retention time for radioac-
tive decay within each trophic level has be-
come less. The effect is thus cumulative along
the food chain.
Figure 6
SEASONAL VARIATIONS IN THE CONCENTRATION OF
P3Z IN COLUMBIA RIVER PLANKTON AND FISH
AUG SEP OCT NOV DEC
Because of the rapid exchange of some nuclides
between the water and algal cells, the activity
density of these organisms will rapidly reflect
changes in the concentration of the radionuclides
in the water. However, the uptake and elimi-
nation of most nuclides in the larger forms,
such as the fish, is a comparatively slow
VI 4-8
-------
Behavior of Radionuclides in Food Chairs - Fresh Water Studies
process. Their activity density will then
fluctuate in a rather sluggish manner in com-
parison to changes in the water or in their
food. Figure 7 shows that trout fed a ration
of P32 each day required about 2 weeks to
reach an equilibrium level. The activity
density of the fish may not therefore, provide
a good indication of the conditions in the
water at the time fish are collected. What
may be more significant however, is that
the fish samples may provide a reasonably
good index of the average conditions which
have existed in the water over a period of
hours or even weeks, dependent on the par-
ticular nuclide involved.
Figure 7
UPTAKE AND RETENTION OF P32 BY TROUT TISSUES
-UN, p-r d« ENti OF P32 FEEDING
,1,1,1 . I
0001,
Watson, George, and Hackett, U.S.A.E.G.
Document HW-59500 (1959)
CONCLUSION
Radionuclides which enter fresh water streams
and lakes may enter the food chain and thus
appear in fish and other aquatic forms eaten
by man. The radioactive contamination levels
which are apt to result in the various organ-
isms, are affected by a large number of phy-
sical, chemical, and biological factors. For
radioisotopes of some of the more abundant
essential elements, we can estimate within
broad limits what the maximum levels may
be. The characteristics of fresh water com-
munities vary so greatly between different
localitits however, that the continued release
of substantial quantities of radionuclides must
be carried out with considerable prudence
and with monitoring of the concentrations of
radionuclides which actually result in the
environs at the point of release.
REFERENCES
1 Love, S. K.. "Natural Radioactivity of
Water", Industrial and Engineering
Chemistry, Vol. 43, pages 1541-1544,
1951.
2 Tsivoglou, E. C., Bartsch, A. F.,
Rushing, D. E., and Holaday, D. A.,
"Effects of Uranium Refinery Wastes on
Receiving Waters", Sew. Ind. Wastes,
Vol. 30, No. 8, pages 1012-1027, 1958.
3 Terrill, J. G., Jr., "Radioactive Waste
Discharges from Nuclear Reactors",
Sew. Ind. Wastes, Vol. 30, pages
270-282, 1958.
4 Staff of Hanford Atomic Products Operation,
"Radioactive Waste Management Opera-
tions at the Hanford Works ", Hearings
Before the Special Subcommittee on
Radiation of the Joint Committee on
Atomic Energy, 86th Congress, on
Industrial Radioactive Waste Disposal,
Jan. 28-Feb.3, 1959, Vol. 1, pages
171-427.
5 Struxness, E. G., Morton, R. J., and
Straub, C. P., "Disposal of High Level
Radioactive Liquid Wastes in Terrestrial
Pits", International Conference on the
Peaceful Uses of Atomic Energy, Geneva,
Paper No. 554, 1955.
S Ophel, I. L., and Fraser, C. D.,
"The Chalk River Liquid Disposal Area",
Atomic Energy of Canada Limited,
Document CR-HP-709, Unclassified
June, 1959.
1 Prosser, C. L., Pervmsek, W., Arnold,
Jane, Svihla, G., and Thompkins, P. D.,
"Accumulation and Distribution of Radio-
active Strontium, Barium-Lanthanum,
Fission Mixture, and Sodium in Gold-
fish", USAEC Document MDDC-496:
pages 1-39, 1945.
VI 4-9
-------
Behavior of Radionuclides in Food Chains - Fresh Water Studies
8 Tomiyama, Tetuo, Kunio Kobayashi, and
Shinya Ishio, "Absorption of 90Sr(90Y)
by Carp," In Research in the Effects and
Influences of the Nuclear Bomb Test
Explosions II. Published by Japan
Society for the Promotion of Science
pages 1181-1187, 1956.
9 Suyehiro, Yasuo, Shizuo Yoshino,
Yoshikazu Tsukamoto, Motohiko
Akamatsu, Kozo Takahashi, and
Takajiro Mori, "Transmission and
Metabolism of Strontium 90 in Aquatic
Animals", In Research in the Effects
and Influences of the Nuclear Bomb
Test Explosions II. Published by
Japan Society for the Promotion of
Science, pages 1135-1141, 1956.
10 Lovelace, F. E., and Podolaik, H. H.,
"Absorption of Radioactive Calcium
by Brook Trout", The Progressive
Fish-Culturist, Vol. 14, pages 154-
158. 1952.
11 Rosenthal, H. L., "Uptake and Turnover
of Calcium-45 by the Guppy", Science
Vol. 34, pages 571-574, 1956.
12 Tomiyama, Tetuo, Shinya Ishio, and
Kunio Kobayashi, "Absorption of Dis-
solved 45Ca by Carassius auratus".
In Research in the Effects and Influences
of the Nuclear Bomb Test Explosions II.
Published by Japan Society for the
Promotion of Science, pages 1151-1156,
1956.
13 Tomiyama, Tetuo, Kunio Kobayshi, and
Shinya Ishio, "Absorption of 32P 64
Ion by Carp", In Research in the Effects
and Influences of the Nuclear Bomb
Test Explosions II. Published by Japan
Society for the Promotion of Science,
pages 1195-1200, 1956.
14 Pendleion, R. C., "Effects of Some En-
vironmental Factors on Bioaccumula-
tion of Cesium-137 in an Aquatic Com-
munity", Biology Research Annual Re-
port - 1958, HW-59500, Unclassified
1959.
15 Watson, D. G.. George, L. A., and
Hackett, P. L., "Effects of Chronic
Feeding of Phosphorus-32 on Rainbow
Trout", Paper Presented at the Inter-
national Congress of Radiation Research.
Burlington, Vermont, August 10-16,
1958.
16 Schiffman, R. H., "The Uptake of Stron-
tium from Diet and Water by Rainbow
Trout", Biology Research Annual Re-
port - 1958 HW-59500, Unclassified,
1959.
17 Vinogradov, A. P., "The Elementary
Chemical Composition of Marine Organ-
isms", Sears Foundation for Marine
Research, Memoir No. 2 (Yale
University, New Haven, Connecticut,
1953).
18 Davis, J. J., Perkins, R. W., Palmer,
R. F., Hanson. W. C., and Cline, J. F.,
"Radioactive Materials in Aquatic and
Terrestrial Organisms Exposed to
Reactor Effluent Water", Second Inter-
national Conference on the Peaceful
Uses of Atomic Energy, Paper No. 393,
1958.
19 Pendleton, R. C., and Hanson, W. C.,
"Absorption of Cesium-137 by Compon-
ents of an Aquatic Community", Second
International Conference on the Peace-
ful Uses of Atomic Energy, Paper No.
392. 1958.
20 Coffin. C. C., Hayes, F. R., Jodrey,
L. H., and Whiteway, S. G., "Exchange
of Materials in a Lake as Studied by
RadioactivePhosphorus", Nature Vol.
163, pages 963-964, 1949a.
21 Coffin, C. C., Hayes, F. R., Jodrey,
L. H., and Whiteway, S. G., "Exchange
of Materials in a Lake as Studied by the
Addition of Radioactive Phosphorus",
Can. J. Res., Sect. D. Vol. 27, pages
207-222, 1949b.
22 Harris, E., "Radiophosphorus Metabolism
In Zooplankton and Microorganisms",
Can. J. Zool. Vol. 35, pages 769-782,
1957.
VI 4-10
-------
Behavior of Radionuclides in Food Chains - Fresh Water Studies
23 Hayes, F.R., McCarter, J.A., Cameron,
M.L., and Livingstone, D. A., "On
the Kinetics of Phosphorus Exchange
in Lakes", Jour. Ecol. Vol. 40, pages
202-216, 1962.
24 Hayes, F.R., Reid, B.L., and Cameron,
M. L., "Lake Water and Sediment. II.
Oxidation-Reduction Relations at the
Mud-Water Interface", Limnol. &
Oceanog. Vol. 3, pages 308-317, 1958.
25 Hutchinson, G. E., and Bow en, V.T.,
"A Direct Demonstration of the Phos-
phorus Cycle in a Small Lake", Proc.
Natl. Acad. Soc. Vol. 33, pages 148-
153, 1947.
26 Hutchinson, G. E., and Bow en, V.T.,
"Limnological Studies in Connecticut.
IX. A Quantitative Radiochemical
Study of the Phosphorus Cycle in
Linsley Pond", Ecology, Vol. 31,
pages 194-203, 1950.
27 McCarter, J.A., Hayes, F.R., Jordrey,
L.H., and Cameron, M.L., "Movement
of Materials in the Hypolimnion of a
Lake as Studied by the Addition of
Radioactive Phosphorus", Can. J.
Zool. Vol.' 30, pages 128-133, 1952.
28 Macpherson, L. B., Sinclair, N.R.,
and Hayes, F.R., "Lake Water and
Sediment. III. The Effect of pH on the
Partition of Inorganic Phosphate
Between Water and Oxidized Mud or its
Ash", Limnol. & Oceanog. Vol. 3,
pages 318-326, 1958.
29 Rigler, F. H., "A Tracer Study of the -
Phosphorus Cycle in Lake Water",
Ecology Vol. 37, pages 551-562, 1956.
30 Whittaker, R.H., "Removal of Radio-
phosphorus Contaminant from the Water
in an Aquarium Community", (Hanford
Works) Biology Research - Annual
Report 1952: 14-19, U.S. Atomic
Energy Commission Document
HW-28636. Unclassified, 1953.
31 Whittaker, R.H., "An Experiment on the
Relation of Phosphate Level in Water
to the Removal and Concentration of
Radiophosphorus", (Hanford Works)
Biology Research - Annual Report
1953: 16-23, U.S. Atomic Energy
Commission Document HW-30437,
Unclassified 1954.
32 Odum, E.P., "Fundamentals of Ecology, "
W.B. Saunders Company, pages 1-546,
1959.
This outline was prepared by R. F. Foster,
Manager, Aquatic Biology, Biology Operation,
Hanford Laboratories, General Electric
Company, Richland, Washington. The report
is based on work performed under contract
No. AT(45-D-1350 for the U.S. Atomic
Energy Commission.
Additional plates by L. G. Williams, Former
Aquatic Biologist, Aquatic Biology Section,
Basic and Applied Sciences Branch. Division
of Water Supply and Pollution Control.
VI 4-11
-------
FWPCA RESPONSIBILITIES FOR WATER QUALITY STANDARDS
I INTRODUCTION
The Water Quality Act of 1965 provided for
the most forceful program yet devised to
protect and enhance the quality of the Nation's
waters. Under this law, the States had until
June 30, 1967, to adopt- water quality criteria
for their interstate and coastal waters and a
plan for implementing and enforcing the
criteria adopted. Criteria and plans acceptable
to the Secretary of the Interior become the
water quality standards applicable to the
State's interstate and coastal waters.
In the event any State standards are unacceptable,
the Secretary of the Interior has the authority
to establish standards, but only after the
States and all other affected interests have
had a full opportunity to be heard. Once
adopted, standards are enforceable by the
States and the Federal Government if any
discharge of material occurs which reduces
the quality of the water below the established
standards.
The standards make it possible for munic-
ipalities, industries, and other water users
to know in advance what their responsibilities
are for keeping clean waters clean and for
restoring polluted waters to a reasonable
degree of purity. The standards also give
the Federal Government authority to prevent
pollution before it occurs, instead of instituting
enforcement action after health and welfare
are proven to be endangered.
The standards are not intended to "lock in"
present uses of water, nor to exclude uses
not now possible. Nor are they intended to
be least common denominators of water
quality and use. Instead, they are intended
to enhance the quality and productivity of the
Nation's water resources in an orderly,
programmed manner.
Standards prepared by the States were sub-
mitted and reviewed by the Administration
prior to submitting them to the Secretary.
The standards-setting effort is the Federal
Water Pollution Control Administration's
top priority program.
When standards are reviewed, the respective
types of water uses determined to be
desirable are evaluated against the water
quality criteria prescribed to achieve those
uses. Close attention is also paid in the
review to plans prepared for implementing
the standards and to surveillance programs
developed for assessing effectiveness of the
proposed program.
To assist States in delineating factors to be
considered in reviewing standards, the
Federal Water Pollution Control Administra-
tion prepared and widely distributed
"Guidelines for Establishing Water Quality
Standards for Interstate Waters" and
"Necessary Supporting Material and
Implementation Plan Contents. " To assist
him in evaluating State submissions, the
Secretary of the Interior appointed a variety
of experts to the National Technical Advisory
Committee, with the request that they pre-
pare water quality criteria for water uses
in the area of their competence. Sub-
committees were established to develop
criteria in the following areas: fish, other
aquatic life and wildlife, municipal water
supplies, industrial water supplies,
agriculture, and recreation and aesthetics.
A comprehensive report has been completed
and is available in limited quantities.
Standards submitted for review have
generally been of a high caliber and rep-
resent a substantial Federal-State-local
cooperative effort. The standards-setting
process has resulted in actions not visible
on the horizon initially. Specific programs
with a definite schedule have been prepared
for cleaning up rivers, lakes, streams, and
coastal waters; State laws to permit com-
pliance with the Water Quality Act have been
revised where necessary; and information
previously unavailable or in an unassembled
form has been developed and collected in a
single set of documents available to all groups
with an interest in preserving and protecting
water quality. Most important, however, is
the spirit aroused in the preparation of these
blueprints for clean water. Officials at all
W.Q. STO.4a. 9. 69
VI 5-1
-------
FWPCA Responsibilities for Water Quality Standards
levels have been surprised at the interest
expressed by citizen groups and have themselves
become actively involved in assuring the
program's success.
Significant as they are, standards are basically
a guide for obtaining the longer range objective
of improving and protecting water quality.
Standards run through the clean-up program,
holding together and guiding efforts in planning,
waste treatment works construction, research,
enforcement, pollution surveillance, training,
and technical assistance.
Standards have been partially or wholely
approved for all States, three territories,
and the District of Columbia. Present
efforts are to effect resolution of unaccept-
able portions of the submitted standards
and to develop acceptable anti-degradation
statements. Development of additional and
more definitive standards may be anticipated
as the program progresses.
Additional information on this program may
be obtained from the Regional Coordinator of
Water Quality Standards or the Water Quality
Standards Staff, Office of Program Plans and
Development, Federal Water Pollution Control
Administration, Washington, D. C. 20242.
The act of setting standards or selecting words
or numbers to describe permissible water
quality ranges is obviously not an end to itself.
Surveillance is required to determine if the
standards are being met, and enforcement
may be necessary if cleanup operations
designed to meet standards are not proceeding
on schedule.
The mission of the water quality surveillance
program is:
A To monitor water quality and waste facility
information for compliance or non-
compliance with water quality standards
and to report these findings to those in a
position, to take action.
B To provide the specific information required
to assess pollution problems and to evaluate
the effects of remedial actions by:
1 Acting as a support service to other
FWPCA programs;
2 Providing technical assistance to the
states;
3 Cooperating with other Federal agencies
and states in the collection of pollution-
related information.
C To supply water quality and related
pollution information for basin-wide
water pollution control planning, to
document pollution control violations for
enforcement actions, and to fulfill
responsibilities as assigned to the program.
The highest priorities for all organizational
elements responsible for this program are
the planning, developing, and coordinating
of the surveillance systems necessary to
determine the compliance or noncompliance
of the water quality standards and
implementation plans.
This will require that Headquarters provide
technical guidance to ensure that all data
gathering and evaluation are accomplished
in a uniform and efficient manner.
Particular attention will be given to improving
the systems for the collection of water quality
data and the establishment of an Early Warning
System which identifies trends in water quality
in advance of the potential violation.
It will be the responsibility of Headquarters
to identify water quality surveillance program
activities as they are presently handled in
other programs so that these may be con-
solidated and that sufficient funds and
personnel are allocated from these programs
to adequately continue the level of
surveillance.
Regions will plan and design their surveillance
systems on a major basin basis ensuring that
the information systems output is current
and accurate.
Efforts in information collection, evaluation,
and dissemination must be highly coordinated
with appropriate Federal, state, local, and
private groups or agencies to avoid duplication
of effort, to ensure proper allocation of
resources, and to consider alternative
methods or courses of action.
Each region will provide support for the
National effort in analytical quality control
in sample handling and analysis. All essential
data developed in the region will be put in a
storage and retrieval system that is accessible
to aU of FWPCA.
VI 5-2
-------
FWPCA Responsibilities for Water Quality Standards
The order of priority for work assignments
in this program will be:
A Water Quality Standards
B Enforcement Activities
C Pollution Control Planning
D Waste Facilities Inventories
E Others as Required
The word "surveillance" suggests a possible
observance or inspection of activities.
Surveillance as practiced by the Federal
Water Pollution Control Administration,
however, involves the collecting of discrete
information expressly to define the status of
water pollution control on a point by point
basis so that timely evaluation and remedial
action can be effected.
Water pollution control in the United States
was launched as a permanent, national
program in 1956 with the enactment of Public
Law 84-660, the Federal Water PoUution
Control Act. Some entirely new concepts were
created by the Water Quality Act of 1965 and
the Clean Water Restoration Act of 1966. In
several respects the recent legislation bears
significantly on the water pollution surveillance
activity in the United States.
The Clean Water Restoration Act of 1966
transferred administration of the Oil Pollution
Act of 1924 from the Secretary of the Army
to the Secretary of the Interior. In the process,
the Oil Pollution Act, which was formerly
limited to vessel pollution affecting coastal
navigable waters and the sea within the
territorial jurisdiction of the United States,
was extended to inland navigable waters and
adjacent shorelines subject to pollution from
boats and vessels. This new authority
complements and expands upon previous
responsibilities vested in the FWPCA by
PL 84-660, as amended, for the control of
pollution resulting from land-based sources,
including oil pollution from shore-based
facilities.
II SURVEILLANCE ACTIVITIES
A Indices
The fresh waters of our nation must be put
to various uses. Among these, municipal
water supply has number one priority.
Asa consequence, statistics are kept on
the location, quantity, and type of water
treatment provided by every municipality
in the United States. Similarly,
inventories are maintained of sewerage
systems (including waste treatment
facilities) delineating the populations
served, the type of treatment provided,
and the additional facilities needed.
National data are also collected, evaluated,
and published periodically on the financing
and construction of sewage collection and
treatment facilities and on pollution-
caused fishkills. At present, the major
unknowns, insofar as waste discharges
are concerned, include those associated
with industrial manufacturing, acid mine
drainage, thermal pollution, and irrigation
return flows. In appreciation of this, the
Secretary of the Interior announced
that the FWPCA will initiate an industrial
waste discharge inventory as a continuing
program.
B Water Quality Data
In the final analysis, progress in the field
of water pollution control will be measured
by the presence or absence of desired
levels of quality in the streams and water-
ways of the Nation. Thus, while the
aforementioned statistics are essential.
Federal surveillance resources must be
aimed primarily at monitoring in-stream
conditions for immediate relation to the
established water quality standards --
point by point, stream by stream, and
basin by basin on a day-to-day basis.
To carry out this mission, the FWPCA's
Division of Pollution Surveillance pursues
the following major stream quality oriented
activities via a central facility and regional
laboratories.
VI 5-3
-------
FWPCA Responsibilities for Water Quality Standards
1 Water Quality Compliance System -
Flexible basin-wide systems of water
quality monitoring stations, including
both central laboratory analyses and
in-stream analyses via automatic
equipment.
2 Oil Pollution Act Laboratory Services -
A unit at the FWPCA laboratory in
Cincinnati specializing in techniques
required for the detection and analysis
of petroleum compounds.
3 Specialized Analytical Services and
Instrumentation - Expert analytical
services and specialized analytical
equipment for national consultation
and services and the development,
installation, and operation of appropriate
automatic water quality sensing and
transmitting instruments.
4 Analytical Quality Control - To assess
and assure the reliability of laboratory
and field analytical processes and results,
including the field testing of new laboratory
analytical procedures, and the initiation
of a complementary program with state
and interstate water pollution control
agencies to assure the validity of data
used in litigation.
5 Data Operations, Evaluation, and Control -
To provide and operate a computerized
system for the timely storage, retrieval,
processing, and analysis of necessary
water quality data and related statistics--
including interstate water quality standards
and implementation plans.
C Surveillance Stations
Aside from the many sampling stations
established for limited duration as part
of special studies and investigations,
FWPCA has, since 1956, established
stations at critical points in major basins
of the United States. Each of the basic
stations is being critically reviewed as to
location, collection frequency, and the
parametric analyses which will be required
in determining compliance with specific
water quality standards. Over 100 of the
stations are located at or near international
or interstate boundaries. Other stations
are located at or near the current or
proposed municipal water intakes of such
major cities as Kansas City, St. Louis,
Cincinnati, Washington, Philadelphia,
Buffalo, and New York City. Stipulating
the development of standards aimed at
protecting and enhancing water quality
at water supply intakes, many stations
obviously must also be located on inter-
state streams below major municipal and
industrial complexes.
Under the terms of an interagency agree-
ment with the U. S. Geological Survey,
the Survey will operate these critical
long-term monitoring stations. The
Survey's responsibilities include sample
collection and analysis and entry of the
data in the STORET System. FWPCA's
responsibilities include continuing review
and evaluation of the data to determine
adequacy of station location and parameters,
frequency of sampling, and the detection
of trends or standards violations.
1 Number - It is currently impossible to
forecast the eventual requirements for
either state or Federal pollution oriented
monitoring. We do know that the number
of stations must be substantially
increased in each major basin to fulfill
the Federal responsibility as well as
give technical assistance to the states
and to permit the Secretary to act
quickly if a state fails to establish or
maintain satisfactory interstate water
quality standards. As in the case of the
basic surveillance stations, all new
stations including reactivation of
stations occupied for short periods
during previous investigations, will
be scrutinized to assure that they
conform to certain basic criteria.
2 Locations required - The locating of the
Federal water quality surveillance
stations will be influenced by the state
implementation plans. However, it is
quite predictable that Federal stations
will be located at the mouths of rivers
and tributaries, the points at which
streams cross state lines, points above
and below major municipal and industrial
complexes, and points designated for
specific water uses, including water-
based recreation and propagation of
fish and wildlife. Some station loca-
tions may be thought of as permanent
VI 5-4
-------
FWPCA Responsibilities for Water Quality Standards
installations inasmuch as it may be
impossible to predict the time when
data will not be needed for that point.
Conversely, a significant number of
stations may measure progress toward
water quality objectives which can be
met within a predictable, definite period
of time. For example, a station
established to monitor a point downstream
from a specific waste problem may well
be deactivated, or at least curtailed,
once the problem is ameliorated.
3 Monitoring criteria - In order to maximize
the results of vested resources, pollution
control monitoring systems must be
designed for both flexibility and specificity
in regard to sampling locations, analyses
performed, and the frequency of analysis.
Water pollution surveillance systems must
be directed primarily toward detecting
changes in the degree of water pollution
associated with man's activities. These
changes occur with relative rapidity as
compared with changes associated with
most natural phenomena; for example,
the changes occasioned by the passage
of water over or through the ground.
A new industrial complex, a new sewage
treatment plant, or any new water
resources structure can effect rapid
changes in water quality; hence, the
basin system and its component monitoring
stations must be periodically reviewed
to ensure flexibility to respond quickly
to such physical changes. Each year
such reviews will undoubtedly result
in the relocating of a number of stations,
curtailing sampling at some, and expanding
the sampling frequency and diversity of
analyses at others. Thus, each basin
system must be designed to detect and
assist in the control of waste discharges,
to determine the effectiveness of waste
treatment and control, and to measure
pollution abatement progress.
4 Parameters required - Although the
adoption of an identical, inviolate list
of parameters to be associated with
each surveillance point would simplify
the administration of each basin
surveillance system, such rigidity
would be neither useful nor desirable.
Surveillance for water pollution control
purposes will be oriented to specifically
stated water quality criteria. Water
pollution problems of varying severity
exist in most rivers and streams of the
United States; yet they are never
identical. The surveillance program
will be tailored to the very special
set of circumstances and needs associ-
ated with the specific problem at
individual locations. In this sense, the
water quality criteria received from the
states show a diversity of parameters
and serve to define those which are
particularly significant for major
reaches of interstate waterways. Thus,
the parameters requiring analyses may
range both within a basin and on a
national basis. They may vary from
readily obtainable onsite information on
dissolved oxygen, conductivity, pH, and
temperature to the sophisticated
analyses of numerous biological entities,
a growing number of radionuclides,
toxic substances such as arsenic and
heavy metals, plus a formidable array
of bacteria, viruses and individual
organic contaminants including pesticides
and petroleum-based materials, all
requiring detailed laboratory evaluation.
5 Frequency of analysis and automatic
equipment - The frequency of sampling
cannot be irrevocably fixed for an entire
system of surveillance stations. Indeed,
it cannot be fixed permanently for a
single station. Periodic adjustments
of frequencies will be necessary for a
variety of reasons. The lowest sampling
frequency capable of providing desired
information is the best frequency.
Automatic sensors can facilitate the
collection of large amounts of data at
closely spaced time intervals and can
be used advantageously for a variety of
purposes. There is a definite place for
such equipment, and there will be more
need for it as the water quality standards
are implemented. However, each
situation will be examined critically to
determine if instrumentation is really
needed. For example, many physical
and chemical characteristics of surface
water remain relatively constant, or
VI 5-5
-------
FWPCA Responsibilities for Water Quality Standards
change slowly. For such parameters,
laboratory analyses of periodic grab
samples often are sufficient. Conversely,
a critical water quality parameter, such
as dissolved oxygen, which is vital to
fish and other aquatic life as well as
being generally indicative of water quality
conditions, may vary under certain
conditions from an acceptable to a non-
acceptable value in a short time. In
general, instrumentation will be applied
only to those situations where the
parameters are subject to quick change,
are essential to the development and
verification of mathematical models for
evaluating alternate pollution control
programs within a basin, or are required
to "sound an alarm" at a critical location.
The guiding principles in selecting
automatic instrumentation, as related to
water pollution control, are presented
in a "Program Guide for Automated
Instrumentation for Water Pollution
Surveillance."
III WATER QUALITY MONITORING ACTIVITIES
OF THE STATES
Consideration of water quality monitoring
activities which can best be conducted by the
states, indicates at least three levels of
surveillance to be satisfied. First, on-site
inspection of municipal and industrial waste
handling procedures must be recognized as a
basic state or local responsibility; this will
assure both more efficient operation of treat-
ment and control facilities and the highest
level of return per unit of regulatory effort.
Second, monitoring of individual plant effluent
lines should be an important part of state
programs. Third, the states should examine
the receiving water body to assure attainment
of desired water quality levels consistent with
adopted water quality standards. The latter
will require the utmost care to assure
coordination and cooperation among the states
and the Federal government in fulfilling their
mutual responsibilities. Indeed, the need for
coordination to assure wise programming of
the limited resources of the responsible
agency is FWPCA's main reason for not
defining the specific stations to be included
in each of its basin monitoring systems.
IV ANA LYTICA L QUA LITY CONTROL
A Need
A principal concern of the FWPCA is the
reliability of water quality measurements
used to determine compliance with water
quality standards. When violations occur,
these data will be used to defend litigation
proceedings. In anticipation of such
circumstances, new emphasis is being
given to analytical quality control within
FWPCA. The agency and those who work
with it must expect court challenges by
those who carry the burden of defense.
Data which cannot be successfully defended
under spirited and argumentative challenges
will mean serious loss of state and Federal
program effectiveness. Thus, the Federal
Water Pollution Control Administration is
taking a strong position toward upgrading
the present system for analytical quality
control and methods validation; before
using information from other sources the
Administration will scrutinize the data--as
it will screen its own--to assure the
successful presentation of the facts
associated with a specific pollution
situation.
V DATA HANDLING
A Coordination
Vital to the FWPCA mission and essential
to a coordinated and cooperative state/
Federal pollution control effort is a unified
or compatible approach to the processing
and timely reporting of surveillance data.
This includes both water quality information
and facilities statistics. Data of the highest
scientific accuracy and precision are of
little value unless they can be applied at
the right time.
VI 5-6
-------
FWPCA Responsibilities for Water Quality Standards
B STORET
1 Computer center - In the past, various
computer systems have been used to
handle the volume of information which
. must flow among the responsible pollution
control agencies. To assist this operation,
FWPCA has established the STORET
system (STOrage and RETrieval) to
assure that all collected information
is available in a timely and concise
fashion. This system has been converted,
along with subsystems now being
developed, for use at the Department of
the Interior's computer center which
, utilizes the IBM 360/65.
2 Extent of use - All FWPCA water quality
data and related information are currently
being placed in the STORET system so
that they can be rapidly retrieved to
support all operating arms of the
Administration throughout the United
States. FWPCA invites the use of the
STORET system by all water pollution
control agencies having need of such
data, and FWPCA frequently inserts
information developed by other Federal
and state agencies if it has a direct use
consistent with the pollution control
mission. At the present time, STORET
contains water quality data and, in many
instances, flow data for over 6, 000
stations which have been operating for
one year or more. At least 3, 500
station records include flow data and/or
quality data collected by the U. S.
Geological Survey which are being used
in enforcement, comprehensive planning,
and water quality standards development
activities of FWPCA. Similarly, FWPCA
projects have inserted data from ten
states, as collected by state water
pollution control or water resource
agencies. In addition, water quality
and quantity data for over 100 locations
have been inserted in STORET to assist
the Tennessee Valley Authority to
implement pollution control responsibilities.
3 Retrieval of data - Since statistical
, printouts of STORET data for specific
locations are available upon request,
many states are now tapping the STORET
reservoir of information. The
availability of concise printouts,
including all available data of reputable
quality at a specific location, is
particularly valuable in judging whether
a proposed criteria will in fact result
in water quality enhancement. At
present, turn-around time for variable
retrievals from the system, as offered
to the various agencies, is dependent
on the mails. Ultimately, FWPCA is
planning a national communications
data handling system which would
further reduce the time required to
respond to requests for services. The
cost of such a system will be small as
compared to the cost of developing
analytical results and will be minor
when one considers the estimated 20 to
50 billion dollar cost of the remedial
construction program. By connecting
the central computer with terminal
facilities located in FWPCA regional
offices, laboratories, and other major
program offices, it will be possible to
utilize the full power of the STORET
system. Such a network will make
STORET and the developing subsystems
available to all elements of FWPCA and
other water pollution control agencies.
VI SUMMARY
In this new era of water pollution control in
the United States, the fight to restore water
quality in our environment has just begun.
Accordingly, the necessary operations are
still in the formative stages as related to the
ultimate needs. The type of information to
be gathered must be carefully selected; the
data must be verified to assure that they meet
their intended purpose. The coordinated
exchange of information among cooperating
and responsible agencies will play a major
role in minimizing the cost of surveillance
activities as well as speeding water pollution
control toward its ultimate goal--enhancement
of the quality and value of the Nation's
water resources.
This outline was prepared for the Training
Program by M. L. Wood, Director, Robert
S. Kerr Water Research Center, Ada,
Oklahoma.
VI 5-7
-------
MARINE AND ESTUARINE PLANKTON
I INTRODUCTION
In recent years the rate of increase in the
human population of the earth and the resulting
need for more food has intensified our interest
in, and increased emphasis on, studies of those
organisms forming the key units or "links" in
oceanic food chains, the plankton. In addition
to this interest, the value of these planktonic
organisms as "indicators" of the origin of
water masses, both oceanic and estuarme,
has become increasingly evident to students of
oceanography and estuarme hydrography.
The common organisms making up the estuarme
and marine plankton and the general environ-
mental factors influencing their occurrence,
abundance and distribution are, therefore,
worthy of the attention of all persons interested
in the biology of these environments.
II THE ESTUARINE ENVIRONMENT
A Definition
An estuary may be variously defined. For
biological purposes it might best be con-
sidered as a region v/here river water
mixes with, and measurably dilutes sea
water; or, as those water masses which,
by virtue of their position, are directly
subject to the combined action of river and
tidal currents.
B General Structure
1 Physical, chemical and environmental
characteristics of an estuary are depend-
ent on the degree of mixing and on the
currents and circulation patterns set up
when two dissimilar water masses meet.
2 Types of estuary:
a Two-layered
b Partially mixed
c Completely mixed
The mixing and circulation of estuaries
are controlled by such varied factors as
wind, rate of fresh-water run-off, tidal
amplitude and physiographic features
such as the slope, width and depth of the
estuarme basin.
These diverse factors acting simultane-
ously make each estuary a unique con-
stantly changing environment.
Ill THE ESTUARINE PLANKTON
A Plankton Types
1 Permanent estuarme plankton is com-
posed of those organisms remaining for
an appreciable Jength of time within the
estuary.
2 Transient estuarme plankton is made up
of those transitory forms from:
a Fresh water feeding into the head of
the estuary.
b Marine waters brought into the
estuary from the sea.
The stay of transient forms in the
estuary is brief and may be limited to
a single tidal cycle.
B Permanent Estuarme Plankton
1 Example: St. John River, New
Brunswick, Canada
a Salt water penetrates 25 miles up-
stream from the river mouth.
BI.MAR. mic. 2a.4.70
VI 6-1
-------
Marine and Estuanne Plankton
1) At the upper end of the estuary
barnacle nauplu and Sagitta elegans
are found in August.
2) Both of these planktomc forms are
less common near the mouth of the
estuary.
b This is an instance where the length
of the estuarine system leads to a
water mass and its plankton becoming
"stranded" in the system and, there-
fore, more or less permanent.
2 Example: Miramichi River, New
Brunswick, Canada
a Smelt larvae occur in samples from
the middle part of the estuary begin-
ning in June.
b During the ensuing summer (June-
August) catches in the same area
indicate growth of the larvae.
c All evidence indicates a light-
controlled diurnal vertical migration
of the larvae. In the summer months
this would keep them in the bottom
layer of inflowing sea water most of
the time.
C Transient Estuarine Plankton
1 Example: Margaree River, Nova
Scotia, Canada
a This is a very short river as com-
pared with the St. John and Miramichi.
b Plankton tows reveal organisms of
marine or freshwater origin depend-
ing on state of the tide.
1) At high tide marine cladocerans,
copepods, ctenophores, polychaete
larvae and marine diatoms are
found.
2) At low tide very few planktomc
organisms are found and these are
all of fresh water origin; e.g.,
Dephnia, Cyclops and freshwater
diatoms.
No permanent plankton exists in the
Margaree estuary, since it is a short,
shallow system.
IV THE MARINE ENVIRONMENT
A When compared with that of the estuary the
environment of the ocean is found to be
more stable.
B Currents
1 No part of the ocean is completely
stagnant. Circulation has evolved in
all oceans, determined largely by the
rotation of the earth, positions of the
sun and moon, the winds and the con-
figurations of the ocean basins.
2 There are also currents caused by
difference in density, by up well ings
and by the entry of river waters into
the sea.
C Planktomc distribution is largely a function
of temperature, nutrients, salinity, light,
and current patterns.
V THE MARINE PLANKTON
A Types of Marine Plankton
1 The phytoplankton includes all the freely
floating photosynthetic forms. Individu-
ally they are microscopic in size and
are unicellular or in groups of cells
loosely bound together.
2 The zooplankton includes a great array
of micro- and macroscopic free floating
animals ranging in size from several
micra to about 50 mm. (Some jellyfish
may reach 1 m. in diameter.)
3 Marine plankton may be further divided
into:
a Temporary plankton consisting of
transitory, floating eggs, larvae and
juveniles of bent hi c and nektomc
organisms, medusoid stages of
benthic coelenterates, and the spores
and gametes of various benthic algae.
VI 6-2
-------
Marine ana Estuanne Plankton
1) Temporary plankton is often
seasonal in occurrence and
generally is confined to neretic
or coastal waters.
b Permanent plankton consists of
organisms completely adapted to
open ocean existence which remain
floating or feebly swimming through-
out their entire life cycle.
B Organisms of the Phytoplankton
1 Diatoms (Bacillanophyceae) are first
in rank as producers of organic material
(especially in northern waters).
a Reproduction
Reproduction in diatoms is by binary
fission. This potential for geometric
increase in population size contributes
to "blooms" under favorable
conditions.
2 Dmoflagellates are generally considered
second to diatoms as primary producers
though often they are more important in
southern waters.
a Dmoflagellates are most well known
for their "blooms" which result in
catastrophic mortalities. Red tides
caused by various species of Gym-
nodmium in southern areas are a
good example.
3 Coccolithophores constitute major com-
ponents of the plankton in some areas.
They are of special geological signifi-
cance because of the accumulation of
their calcium carbonate coccoliths in
marine sediments.
C Factors Affecting Distribution of
Phytoplankton
1 Light
The amount of light energy available to
photosynthesizers diminishes with depth.
2 Vertical circulation
Adequate supplies of plant nutrients
(especially nitrates and phosphates)
must be present if the phytoplankton
is to grow.
a The most abundant source of such
nutrients is bottom sediment which
is out of reach of the photosynthesizing
organisms.
b Methods of returning nutrients to the
surface are:
1) Upwellings (most commonly en-
countered near shore).
2) Divergences of major currents
(more common in areas far from
shore).
3 Temperature and salinity act largely
as selective agents.
D Marine Zooplankton
1 The marine zooplankton contains organ-
isms in every major group of marine
animals except the sponges, bryozoans,
brachiopoda, and mammals. Copepods
are the most abundant representatives.
2 The greatest significance of the zoo-
plankton lies in their role as organic
links in the biological economy of the
sea.
3 Cyclic Representation of a General
Food Chain in the Sea:
Sun
CO,
\
PH
PHYTOPLANKTON ) GRAZERS
\
Nutrients
CARNIVORES I
Death &'
Sedimentation
I
vDeath &
Excretion* CARNIVORES II
VI 6-3
-------
Marine and Estuarine Plankton
REFERENCES
1 Hurt, W.V. and McAlister, W. B. Recent
Studies on the Hydrography of Oregon
Estuaries. Res. Briefs, Fish. Comm.
of Oregon, Vol. 7, pp. 14-27. 1959.
2 Clemens, W.A. Pastures of the Sea.
Occasional Pap. Calif. A cad. Sci.,
No. 41, 3 figs., 8 pp. 1963.
3 Hardy, A. C. The Open Sea, Its Natural
History: The World of Plankton.
Houghton-Mifflin Co., Boston, 335 pp.
1956.
4 Johnson, M. W. Plankton. IN J.W.
Hedgpeth, ed., Treatise on Marine
Ecology and Paleoecoplogy, Vol. 1,
Ecology. Chap. 16, pp. 443-459, Mem.
67, Geological Soc. of America. 1957.
5 Ketchum, B. H. The Exchanges of Fresh
and Salt Waters in Tidal Estuaries.
Jour. Marine Res., Vol. 10, pp. 18-
38. 1951.
Olson, Theodore A. and Burgess,
Frederick J. Pollution and Marine
Ecology. Interscience Publishers.
364 pp. 1967.
Rogers, H.M. Occurrence and Retention
of Plankton within the Estuary. Jour.
Fisheries Res. Bd. Canada, Vol. 5,
pp. 164-171. 1940.
This outline was prepared by G. C. Hughes,
Assistant Professor of Biological Sciences,
University of the Pacific, and Assistant
Professor of Marine Science, Pacific
Marine Station, Dillon Beach, Marin County,
California.
VI 6-4
-------
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
organisms 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, Lais on, 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 A djective
various epiholitic, nereiditic, sessile
plants epiphytic
animals epizooic
wood epidendritic, epixylonic
rock epihthic
[After Srameck-HuseM 1946) and via Sladeckova
(1962)] Most above listed latin-root adjectives
are derivatives of nouns such as epihola,
epiphyton, spizoa, etc.
BI.MIC.enu. 19a.4. 70
VI 7-1
-------
Attached Growths (Periphyton or Aufwuchs)
III Periphyton, as with all other components
of the environment, can be sampled quali-
tatively (what is present) and quantitatively
(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 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
serve as the principal periphyton
substrates.
V ARTIFICIAL SUBSTRATE PLACEMENT
A Position or Orientation
1 Horizontal - Includes effects of settled
materials.
2 Vertical - Eliminates many effects of
settled materials.
B 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.)
C Current is Important
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 d. e. detritus, sub-
strate, etc.) so that some commonly A
employed quantitative procedures are
not applicable.
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
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).
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.
VI 7-2
-------
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.
VIII DETERMINING THE QUANTITY OF
GROWTH(S)
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.
4 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)
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
K EXPRESSION OF RESULTS
A Qualitative
1 Forms found
2 Ratios of number per group found
3 Frequency distribution of varieties
found
B Quantitative
1 A real 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 O?/mg of growth/hour
VI 7-3
-------
Attached Growths (Periphyton 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 Periphyton. 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 Periphyton Community.
Botanical Review. 28:2:286. 1962.
Srameck~Husek (On the Uniform
Classification of Animal and Plant
Communities in our Waters).
Sbornik MAP 20:3:213. Orig. in
Czech. 1946.
Thomas, N.A. Method for Slide
Attachment in Periphyton Studies.
Manuscript. 1968.
Weber, C.I. Methods of Collection and
Analysis of Plankton and Periphyton
Samples in the Water Pollution
Surveillance System. Water Pollution
Surveillance System Applications and
Development Report No. 19, FWPCA,
Cincinnati. 19+pp. (multilith). 1966.
This outline was prepared by Lowell E. Keup,
Acting Supervisory Biologist, Biological and
Chemical Section, National Field Investigations
Center, FWPCA, U. S. Dept. of the Interior,
Cincinnati, OH.
VI 7-4
-------
ARTIFICIAL AND RELATED SUBSTRATES - REFERENCES
INTRODUCTION
This reference list in the field of artificial
substrates has been prepared for those not
familiar with the literature and for those
planning to use one or more of these
techniques. It includes references both to
periphyton and benthos including techniques,
rationale, and practical applications. Since
1967, Dr. C. I. Weber of the Analytical
Quality Control Laboratory has compiled an
annual bibliography for the Midwest
Benthological Society. These bibliographies
are recommended for additional and current
references. This list is thus complementary
to Weber's annual citations for the periphyton.
Addresses and sources are given where
possible.
Priority has been given to comprehensive
works; those of historical note, especially
where the original description of a sampler
is given, and a sampling of recent papers
to show versatility and application for general
and specific uses.
GENERAL
Cooke, William B. Colonization of Artificial
Bare Areas by Microorganisms. Bot.
Rev. 22(9):613-638. Nov. 1956.
Sladeckova, Alena. Limnological Investiga-
tion Methods for the Periphyton C'Aufwuchs")
Community. Bot. Rev. 28(2):286-350.
1962.
Cummins, K. W. An Evaluation of Some
Techniques for the Collection and Analysis
of Benthic Samples with Special Emphasis
on Lotic Waters. American Midland
Naturalist, Vol. 67, No. 2. pp. 477-504.
1962.
PERIPHYTON - GENERAL
Weber, C. E. (FWPCA, 1014 Broadway
Cincinnati, OH 45202) Benthic Macro-
invertebrates and Periphyton. Select and
Current Bibliographies. Midwest
Benthological Society.
Sladeckova, Alena. The Significance of the
Periphyton in Reservoirs for Theoretical
and Applied Limnology. Verh. Internat.
Verein. Limnol. 16:753-758. 1966
PERIPHYTON - PARAFFIN-COATED
SUBSTRATES
Beers, G. D. and Neuhold, J. M.
Measurement of Stream Periphyton on
Paraffin-Coated Substrates. Limnol.
andOceanogr. 13(3)-559-562. 1968.
PERIPHYTON - GLASS SLIDES
See general references above.
Thomas, Nelson A. (National Center for
Field Investigations, FWPCA, 5555
Ridge Avenue, Cincinnati, OH 45213. )
Method for Slide Attachment in Periphyton
Studies. Manuscript. (Nonhardening
"Plasti-tak" used for attaching slides to
bricks. )
Weber, Cornelius I., and McFarland,
Ben H. (FWPCA, Analytical Quality
Control, 1014 Broadway, Cincinnati, OH
45202) Periphyton Biomass-Chlorophyll
Ratio as an Index of Water Quality. 17th
Annual Meeting, Midwest Benthological
Society. 19 pp. Multilithed. 1969.
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.
PERIPHYTON - DIATOMS
Patrick, R , Hohn, M. H., and Wallace, J.H.
A New Method for Determining the Pattern
of the Diatom Flora. Notulae Naturae,
Academy of Natural Sciences of Philadelphia.
No. 259. pp. 1-12. 1954.
Hohn, M. H. The Use of Diatom Populations
as a Measure of Water Quality in Selected
Areas of Galveston and Chocolate Bay,
Texas. Publications of the Institute of
Marine Science of the University of Texas.
Vol. 6. pp. 206-212. 1959.
Patrick, R. Factors Affecting the Distribution
of Diatoms. Botanical Review. Vol. 14.
No. 8. pp. 473-524. 1948.
Patrick. R. A Discussion of Natural and
Abnormal Diatom Communities. In:
Jackson, D. F. (ed.), Algae and Man.
Plenum Press, New York. pp. 185-204.
1964.
BI. BIB. 1.5.70
VI 8-1
-------
Artificial and Related Substrates - References
Hohn, M. Determining the Pattern of the
Diatom Flora. Journal of the Water
Pollution Control Federation, Vol. 33.
No. 1. pp. 48-53. 1961.
Hohn, M. The Relationship Between Species
Diversity and Population Density in Diatom
Populations from Silver Springs, Florida.
Transactions of the American Micro-
scopical Society. Vol. 80. No. 2. pp.
140-165. 1961.
PERIPHYTON - PLASTIC SLIDES AND
PANELS
Grzenda, Alfred R. and Brehmer, Morris L.
A Quantitative Method for the Collection
and Measurement of Stream Periphyton.
Limnol. Oceanog. 5(2):190-194.
PERIPHYTON - DIATOMS - STYROFOAM
Hellerman, J. A Study of the Diatoms of
Mohonk Lake, New York and Vicinity.
Unpublished M. S. Thesis. Rutgers - The
State University. 1962.
Hohn, Matthew H. (Department of Biology,
Central Michigan University, Mt. Pleasant,
Michigan). Artificial Substrate for Benthic
DiatomsCollection, Analysis, and
Interpretation, p. 87-97. 1966. In:
Organism Substrate Relationships in
Streams Pymatuning Special Publication.
No. 4. $2.50. Pymatuning Laboratory
of Ecology, University of Pittsburgh.
Pittsburgh, PA 15213.
PERIPHYTON - TRICKLING FILTERS
Cooke, W. B. Continuous Sampling of
Trickling Filter Populations. Sewage and
Industrial Wastes. Vol. 30. No. 1.
pp. 21-27. 1958.
Cooke, W. B. Fungi in Polluted Water and
Sewage. IV The Occurrence of Fungi in
a Trickling Filter-type Sewage Treatment
Plant. Proceedings of the 13th Industrial
Waste Conference, Purdue University,
Series No. 96, Vol. 43. No. 3. pp. 26-45.
1959.
Cooke, W. B. Trickling Filter Ecology.
Ecology. Vol. 40. No. 2. pp. 273-291.
1959.
PERIPHYTON - LABORATORY STUDIES
Ehrlich, Gary G. and Slack, Keith V.
(Water Res. Div., USGS, Menlo Park,
California 94025). Uptake and Assimila-
tion of Nitrogen Microecological Systems.
Spec. Tech. Pub. 448. ASTM. p. 11-23.
1969.
MACROINVERTEBRATES - GENERAL
Macan, T. T. Methods of Sampling the
Bottom Fauna of Stony Streams. Mitt.
Intern. Ver. Limnol. No. 8. p. 1-21.
1958.
MACROINVERTEBRATES - BRUSH BOXES
Scott, D. C. Biological Balance in Streams.
Sewage and Industrial Wastes. Vol. 30.
No. 9. pp. 1169-1172. 1958
MACROINVERTEBRATES - ROCK BASKETS
Anderson, J. B. and Mason, William T., Jr.
A Comparison of Benthic Macroinvertebrates
Collected by Dredge and Basket Sampler.
Jour. Water Poll. Cont. Fed. 40(2):252-259.
Anderson, J. B. and Mason, W. T., Jr.
The Use of Limestone-Filled Samples for
Collecting Macroinvertebrates from
Large Streams. Water Poll. Surv. Syst.
Application and Develop. Report No. 17,
FWPCA-USDI, Cincinnati, Ohio. May 1966.
Henson, E. B., Jr. A Cage Sampler for
Collecting Aquatic Fauna. Turtox News.
43:298-299. 1965.
Hilsenhoff, William L. An Artificial Substrate
Device for Sampling Benthic Stream
Invertebrates. Limnology and Oceanography.
14(3):465-471. 1969.
Mason, W. T., Anderson, J. B., and
Morrison, G. E. (FWPCA, Analytical
Quality Control, 1014 Broadway, Cincinnati,
Ohio) Limestone-filled, Artificial Substrate
Sampler-float unit for Collecting Macro-
invertebrates in large streams. Progressive
Fish Culturist. 29:74. 1967.
Wene, G., and Wickliff. E. L. Modification
of the Stream Bottom and Its Effect on the
Insect Fauna. Can. Entomologist. 72:131-
135. 1940.
VI 8-2
-------
Artificial and Related Substrates - References
MA GROIN VERTEBRATES - MULTIPLE
PLATES
colonized "than the living plant"
(invertebrate s).
Arthur, JohnW. and Horning, William B.,
(Federal Water Pollution Control
Administration, Duluth, Minnesota,
National Water Quality Laboratory.
The Use of Artificial Substrates in
Pollution Surveys. The Amer. Midland
Natur. Vol. 82. No. 1. pp. 83-89. 1969.
Hester, F. E. and Dendy, J S.
A Multiple-Plate Sampler for Aquatic
Macroinvertebrates. Trans. Am. Fish.
Soc. 91 (4):420-421. April 1962.
Brewer, Jesse W. and Gleason, Gale R.
Modification of the Dendy Principle for
Stream Bottom Sampling. Midwest
Benthological Society. 12th Annual Mtg.
Mimeo. 1964.
Dendy, J. S.
Animals.
1963.
Living Food for Aquatic
Turtox News 41(10):258-259.
MACROINVERTEBRATES - CEMENT
PLATES
Britt, N. W. New Methods for Collecting
Bottom Fauna from Shoals or Rubble
Bottoms of Lakes and Streams.
Ecology. 36:524-525. 1955.
MACROINVERTEBRATES - BURIED TRAYS
Moon, H. P Methods and Apparatus Suitable
for an Investigation of the Littoral Region
of Oligotrophic Lakes. Int. Rev. Hydrobiol.
32:319-333. 1935.
MACROINVERTEBRATES - PLASTIC TAPES
FOR BLACKFLIES
Williams, T. R. and Obeng, L. A Compari-
son of Two Methods of Estimating Changes
in Simulium Larvae Populations, with a
Description of a New Method. Ann. Trop.
Med. Parasit. 56:358-361. 1962.
MACROINVERTEBRATES - ARTIFICIAL
WEEDS
Freshwater Biological Association (The
Ferry House. Far Sawrey, Ambleside.
Westmoreland, England). 1969.
Thirty-seventh Annual Report, p. 36.
Artificial weeds made of strands of
polyethylene and polypropylene twine.
'Artificial Littorella is more heavily
MACROINVERTEBRATES - BRICKS
Elvins, B J. Investigation of the Animal
Population in Polluted Streams. Journ.
Inst. Sewage Purif. Part 6. p. 569.
MACROINVERTEBRATES - 10 or 20 Stone
Surveys
Chutter, F. M. On the Ecology of the Fauna
of Stones in the Current in a South African
River Supporting a Very Large Simulium
(Diptera) Population. J. App. Ecol.
5:531-561. 1968.
Williams. T. R and Obeng, L. A Comparison
of Two Methods of Estimating Changes in
Simulium Larval Populations, with a
Description of a New Method. Ann. Trop.
Med. Parasit. 56:358-361. 1962.
Reed, Roger J. 1966. Some Effects of DDT
on the Ecology of Salmon Streams in
Southeastern Alaska. USFWS Special
Scientific Report No. 542. 15 pp.
Cope, Oliver B. Effects of DDT Spraying
for Spruce Budworm on Fish in the
Yellowstone River System. Trans. Amer.
Fish. Soc. 90:239-251.
MACROINVERTEBRATES - ECOLOGY
Waters, T. F. Recolonization of Denuded
Stream Bottom Areas by Drift. Trans-
actions of the American Fisheries Society,
Vol. 93, No. 3, pp. 311-315. 1964.
Waters, T. F. Standing Crop and Drift of
Stream Bottom Organisms. Ecology.
42:532-537. 1961.
Driscoll, E. G. Attached Epifauna-Substrate
Relations. Limnol. Oceanogr. 12(4):633-641.
1967.
MICROINVERTEBRATES - SESSILE
PROTOZOA
Spoon, D. M. and Burbanck, W. D.
A New Method for Collecting Sessile
Ciliates in Plastic Petri Dishes with Tight
Fitting Lids. J. Protozool. 14(4):735-739.
1967.
VI 8-3
-------
Artificial and Related Substrates - References
Burbanck, W. D. (Emory University,
Atlanta, Georgia) 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.
Sickel, James B. A Survey of the Mussel
Populations (Unionidae) and Protozoa of
the Altamaha River with Reference to
Their Use in Monitoring Environmental
Changes. MS Thesis. Emory University.
133 pp. 1969.
Sladecova, Alena. (Inst. Chem. Tech.,
Prague, Czechoslovakia) Factors
Affecting the Occurrence and Stratifica-
tion of Sessile Protozoans in Artificial
Reservoirs, (in Russian.Summ. in English
and Czech) Technology of Water.
8(1):483-490. 1964.
SUBSTRATES - BUOYS AND OTHER
NAVIGATIONAL AIDS
Miller, Milton A. Isopoda and Tanaidacea
from Buoys in Coastal Waters of the
Continental U. S., Hawaii, and the
Bahamas (Crustacea). Proc. U. S. Nat.
Mus. 125 (3652):53 pp. 1968.
Fremling, C. R. Biology and Possible
Control of Nuisance Caddisflies of the
Upper Mississippi River. Agricultural
and Home Economics Experiment Station,
Iowa State University of Science and
Technology. Ames, Iowa. Research
Bulletin 483, pp. 856-879. 1960.
RECOVERY DEVICES
Ziebell, Charles D., McConnell, W J ,
and Baldwin, Howard A. A Sonic
Recovery Device for Submerged Equipment.
Limnol. and Ocean. 13(1):198-200. 1968.
Fox, Alfred C. (Univ. of Georgia.
Cooperative Fisheries Unit, Athens, Georgia)
Personal Communication. Use of Inexpensive
Detonator; "Seal Salute. " Miller Fireworks
TVA, Div. of Health and Safety,
Water Quality Branch. 1967.
Waters, T F Notes on the Chlorophyll
Method of Estimating the Photosynthetic
Capacity of Stream Periphyton. Limnol.
Oceanogr. 6:486-488. 1961.
Wetzel, R. G. Techniques and Problems
of Primary Productivity Measurements
in Higher Aquatic Plants and Periphyton.
In: C. R. Goldman et. "Primary
Productivity in Aquatic Environments. "
Univ. Calif. Press. Berkeley. 1966.
U. S. Dept. of the Interior, FWPCA.
Keup, Lowell and Stewart, Keith.
National Field Investigations Center,
4676 Columbia Parkway, Cincinnati,
OH 45226. Effects of Pollution on
Biota of the Pigeon River, North
Carolina and Tennessee. 35 pp. 1966.
Utilization of glass slides attached
to bricks for periphyton, chlorophyll
biomass ratios to evaluate paper mill
and other industrial effluents.
Keup, Lowell. National Field
Investigations Center. Effects of
Pollution on Aquatic Life Resources
of the South Platte River Basin in
Colorado. Vol. 1 and Vol. 2. 1967.
Vollenweider, Richard A. et al. (Eds.)
A Manual on Methods for Measuring
Primary Production in Aquatic Environ-
ments. International Biological Programme
Handbook. No. 12. 213 pp. Davis. 1969.
NOTE: Mention of commercial products and
manufacturers does not imply endorsement
by the Federal Water Pollution Control
Administration and the U. S. Department of
the Interior.
and Novelty Company,
Holland, Ohio 43528.
shipping. 1969.
501 Gleneary Road,
$4.00 gross and
ARTIFICIAL SUBSTRATES - PRACTICAL
APPLICATIONS IN WATER POLLUTION
CONTROL
Taylor, Mahlon P. Thermal Effects on the
Periphyton Community in the Green River.
VI 8-4
-------
CHAPTER VII
IDENTIFICATION KEYS
Key to Selected Groups of Freshwater Animals
Key to Algae of Importance in Water Pollution
1
2
-------
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, Ib), and select the alternative
that seems most reasonable. If you
selected "la" you have identified the
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.21E.8.69
VII 1-1
-------
Key to Selected Groups of Freshwater Animals
la The body of the organism comprising
a single microscopic independent
cell, or many similar and indepen-
dently functioning cells associated
m a colony with little or no differ-
ence between the cells, i.e. , with-
out forming tissues; or body com-
prised of masses of multmucleate
protoplasm. Mostly microscopic,
single celled animals.
Phylum PROTOZOA
Ib The body of the organism com-
prised of many cells of different
kinds, i.e., forming tissues.
May be microscopic or macro-
scopic.
2a Body or colony usually forming
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
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 of wheel animalcules.
Phylum TROCHELMINTHES
(Rotifera) (Fig. 3)
4b Larger, wormlike, or having
strong skeleton or shell.
5a Skeleton or shell present. Skel-
eton may be external or internal.
5b Body soft and /or wormlike.
Skin may range from soft to
parchment -like.
6a Three or more pairs of well
formed jointed legs present.
Phylum ARTHROPODA (Fig. 4)
6b Legs or appendages, if present,
limited to pairs of bumps or hooks.
Lobes or tenacles, if present,
soft and fleshy, not jointed.
7a Body strongly depressed or
flattened in cross section.
7b Body oval, round, or shaped like
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
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)
lOa 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)
lOb Divided into sections or segments
15
19
10
11
VII 1-2
-------
Key to Selected Groups of Freshwater Animals
lOc Unsegmented, head blunt, one 18
or two retractile tentacles.
Flat pointed, tail.
lla 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 12
jaws (if present). Fig. 8E.)
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 ANNELIDIA (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, nereid worms.
Phylum ANNELIDIA (Fig. 9)
14a Segments with bristles and/or fleshy
lobes or other extentions. 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, often slimey, 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 PELECYPODA (Fig. 10)
19a Three pairs of regular walking 29
legs, or their rudiments. Wings
present in all adults and rudiments
in some larvae.
Class INSECTA (Figs. 22, 24D,
25, 26, 28, 29)
19b More than three pairs of legs
apparently present.
20a Body elongated, head broad and flat
20
'VII 1-3
-------
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 BRANCHIPODA
(Fig. 16)
22b Less than ten pairs of swimming 23
or respiratory appendages.
23a Body and legs inclosed in bi- 24
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.
Cyclops or cladocera.
Subclass CLADOCERA (Fig. 12)
24b Locomotion accomplished by 25
body legs, not by antennae.
25a Appendages leaflike, flattened,
more than ten pairs.
Subclass BRANCHIPODA
(See 22 a)
25b Animal less than 3 mm, in length.
Appendages more or less slender
and jointed, often used for walking.
Shells opaque. Ostracodes.
(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. 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)
VII 1-4
-------
Key to Selected Groups of freshwater Animals
29b
30a
No functional wings, though
pads in which wings are develop-
ing may be visible. Some may
resemble adult insects very
closely, others may differ ex-
tremely from adults.
30
External pads or cases in which 35
wings develop clearly visible.(Figs.
24.16.27)
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
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. 24Q
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
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)
38
VE 1-5
-------
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.
Ofcder COLEOPTBRA
(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).
4Ob No paired appendages. Mouth
a round suction disc.
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
42a Paired appendages are legs 43
42b Paired appendages are fins,
gills covered by a flap
(operculum). 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
42 44a Warm blooded
41 44b Cold blooded. Body covered
with horny scales or plates
Class REPTILIA
45a Body covered with feathers
Birds.
Class AVES
45b Body covered with hair
Mammals.
Class MAMMALIA
45
VII 1-6
-------
Key to Selected Groups of Freshwater Animals^
REFERENCES - Invertebrates
REFERENCES - Fishes
Eddy, S. and Hodson, A. C.
Taxonomic Keys to the Common
Animals of the North Central States"
Burgess Pub. Co. , Minneapolis,
pp 1-141. 1955.
Edmondson, W. T. (ed). and Ward
and Whipple's Freshwater Biology.
John Wiley & Sons, New York.
pp 1-1248. 1959.
Jahn, T. L. and Jahn, F.F. "How
to Know the Protozoa" Win. C.
Brown Company, Dubuque, Iowa.
pp 1-234. 1949.
Kudo, R. "Protozoology" Charles
C. Thomas, Publisher, Spring-
field, Illinois, pp 1-778. 1950.
Palmer, E. Lawrence "Fieldbook
of Natural History" Whittlesey
House, McGraw-Hill Book Co.,
Inc., New York. 1949.
Pennak, R. W. "Freshwater Inverte-
brates of the United States"
The Ronald Press Co. , New York
pp 1-769. 1953
Pr.afo, W. W. 'M MMtunA of tfa* Common
Invertebrate Animals Exclusive of
Insects" The Blaikston Co. , Phila.
pp 1-854. 1951
American Fisheries Society A List
of Common and Scientific Names
of Fishes from the United States
and Canada. Special Publication
No. 2, Am. FishSoc., Dr. E.A
Seaman, Sec. -Treas., Box 483,
McLean, Va (Price $1. 00 paper,
$2. 00 cloth). 1960.
Bailey, Reeve M. A Revised List of
the Fishes of Iowa with Keys for
Identification, IN: Iowa Fish and
Fishing. State of Iowa, Super, of
Printing. 1956. (Excellent
color pictures.)
Eddy, Samuel "How to Know the
Freshwater Fishes" Wm. C
Brown Co., Dubuque, Iowa. 1957.
Hubbs, C. L. and Lagler, K.F.
Fishes of the Great Lakes Region.
Bull. Cranbrook Inst Science,
Bloomfield Hills, Michigan. 1949.
Lagler, K.F.
Biology"
Dubuque,
"Freshwater Fishery
Wm. C Brown Co. ,
Iowa. 1952
Trautman, M. B. "The Fishes of Ohio"
Ohio State University Press,
Columbus 1957. (An outstanding
example of a State study )
VII 1-7
-------
Key to Selected Groups of Freshwater Animals
1. Spongilla spicules
Up to .2 mm. long.
3A Rotifer, Polyarthra
' ~
3B. Rotifer. Keratella
Up to .3 mm.
4A. Jointed leg
Caddis fly
4B. Jointed leg
Crayfish
2B. Bryosoal mass. Up to
several feet diam.
3C- Rotifer, Philpdina
Up to. 4 mm.
2A. Bryozoa, Plumatella. Individuals up
to ? mm. Intertwined masses maybe
very extensive.
4C. Jointed leg
Ostracod
5. Tapeworm head.
Taenia. Up to
25 yds. long
6A. Turbellaria, Mesostoma
Up to 1 cm.
6B. Turbellaria, Dugesia
Up to 1. 6 cm.
7. Nematodes. Free living
forms commonly up to
1 mm. , occasionally
more.
-------
Key to Selected Groups ojJFreshwater Animals
8B. Diptera, Mosquito
pupa. Up to 5mm.
8A. Diptera, Mosquito larvae
Up to 15 mm. long.
8C. Diptera, chironomid
larvae. Up to 2 cm.
ilE'
pupa. Up to 2. 5 cm.
9D. Diptera, Rattailed maggot
Up to 25 mm. without tube
9A. Annelid,
segmented
worm, up to
1/2 meter
10B. Alasmidonta, end view.
10A. Pelecyopod, Alasmidonta
Side view, up to 18 cm. long.
9B. Annelid, leech up to 20 cm.
12A. Branchiopod,
Daphnia. Up
to 4mm.
11A. Ostracod, Cypericus
Side view, up to 7 mm.
HB. Cypericus, end view.
12 B. 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
vn 1-10
19A. Calanoid copepod, ,
Female 19B. Cyclopoid copepodr
Up to 3 mm. Female
Up to 25 mm.
-------
Key to Selected Groups of Freshwater Animals
V"n
21A.
21B.
21C,
21D. 21E.
21. Trichoptera, larval cases,
mostly 1-2 cm.
22. Megaloptera, alderfly
Up to 2 cm.
E/-'
23A. Beetle larvae, 23B. Beetle larvae, 24A. Odonata; dragonfly
Dytisidae, Hydrbphilidae nymph up to 3 or
Usually about 2 cm. Usually about 4 cm
I 1 cm.
ra:
24B". Odonata, tail
of damselfly
nymph
(side view)
Suborder
Zygoptera
(24B, D)
24D. Odonata, damselfly
nymph (top view)
24E, Odonatar front view
///" / of dragonfly nymph
showing "mask"
partially extended
Suborder
Anisoptera
, E, C)
24C. Odonata, tail of
dragonfly nymph
(top view)
VII 1-11
-------
Key to Selected Groups of Freshwater Animals
25A. Hemiptera.
Water Boatman
About 1 cm.
25B. Hemiptera,
Water Scorpion
About 4 cm.
Plecoptera,
Stonefly nymph
Up to 5cm.
27 .Epheme ropte ra,
Mayfly nymph
Up to 3cm.
28A. 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.
Diptera, Mosquito
Up to 20 mm.
VII 1-12
-------
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
1' 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) Calls in isolated or clustered threads or ribbons which are only one cell thick or wide. 5
4J 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' Heterocysta 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 (Cloeotrichia) ..... 9
8' No spore (akinete) present (Rivularia) ......................... .10
9 (8) Gelatinous colony a smooth bead ....................... Cloeotncrua echinulatc
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 (7'1 Cells adjacent to heterocyst wider than heterocyst ....... ... Calothrix braunii
11' Cells adjacent to heterocyst narrower than heterocyst . ... Calothrix parietina
12(6') Branching present
12" Branching absent
13(12) Branches in pairs ............ Scytonema tolypothricoides
13' Branches arising singly. .. .. ---- . Tolypothrix tenuis
14(12') Heterocyst terminal only (Cvclindrospermum) .................................... 15
14' Hetrocysts intercalary (within the filament) ---- ... . 16
15 (14) Heterocyst round . . . . ................ ... Cylindrospermum muscicola
15' Heterocyst elongate ............... . Cylindrospermum stagnate
16 (141) 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 ...... Nodular la spumigena
18' Heterocysts and vegetative cells not shorter than the thread width ................... -19
19(18') Heterocysts rounded (Anabaena) ............................................. 20
19' Heterocysts chndric. ...................................... Aphanizomenon flos -aquae
20(19) Cells elongate, depressed in the middle; heterocysts rare. ........... Anabaena constricta
20' Cells rounded, heterocysts common ............................................... 21
21(20') Heterocysts with lateral extension! ....................... Anabaena planctonica
21' Heterocysts without lateral extensions .............................................. £2
-------
22(21') Threads 4-8|» wide Anabaena flos-ag^ae
22' Threads 8-14M wide Anabaena circinalla
23(5') Branching absent 24
23' Branching (including "false" branching) present 84
24 (23) Cell pigments distributed throughout the protoplasm 25
24' Cell pigments limited to plastids 49
25 (23) Threads short and formed as an even spiral 285
25' Threads very long and not forming an even spiral "
26(25') Several parallel threads of cells in one common sheath Microcoleus subtorulosus
26' One thread per sheath if present 27
27(26') Sheath or gelatinous matrix present 28
27' No sheath nor gelatinous matrix apparent (Oscillatoria) 35
28(27) Sheath distinct; no gelatinous matrix between threads (Lvngbya) 29
28' Sheath indistinct or absent, threads interwoven with gelatinous matrix between (Phormidium). . .
29(28) Cells rounded LynKbYa ocracea
29' Cells short cylindric 30
30 (29') Threads in part forming spirals LYnBbYa lafierheimii
30' Threads straight or bent but not in spirals . 31
31(30') Maximum cell length 3 5n ; sheath thin Lyngbya dipuett
31' Maximum cell length 6 SM ; sheath thick Lyngbya versicolor
32(28') Ends of some threads with a rounded swollen "cap" cell .... .. . . . 33
32' Ends of all threads without a "cap" cell 34
33(32) End of thread (with "cap") abruptly bent . Phormidium uncmatum
33' End of thread (with "cap") straight . Phormidium autumnale
34 (32') Threads 3-5ii in width ... . . ... Phormidium inundatum
34' Threads 5-12M in width. . Phormidium retzu
35 (271) Cells very short, generally less than 1/3 the thread diameter . . .36
35' Cells generally 1/2 as long to longer than the thread diameter 39
36(35) Cross walls constricted Oscillatoria ornata
36' Cross walls not constricted ... 37
37(36') Ends of thread, if mature, curved . . 38
37' Ends of thread straight ... Oscillatoria limosa
38 (37) Threads 10-14|> thick . ... . . . Oscillatoria curvicepa
38' Threads 16-60|i thick Oscillatoria pnnceps
39(35') Threads appearing red to purplish Oscillatoria rubeacena
39' Threads yellow-green to blue-green . . . 40
40(39') Threads yellow-green 41
40' Threads blue-green 43
41 (40) Cells 4-7 times as long as tne thread diameter Oscillatoria put rid a
41' Cells less than 4 times as long as the thread diameter 42
42 (41') Prominent granules ("pseudovacuoles") in center of each cell Oscillatoria lauterbornii
42' No prominent granules in center of cells Oscillatoria chlorina
43(40') Cells 1/2-2 times as long as the thread diameter 44
43' Cells 2-3 times as long as the thread diameter 48
44 (43) Cell walls between cells thick and transparent Oscillatoria pseudogeminata
44' Cell walls thin, appearing as a dark line 45
vn 2-2
-------
45(44') Ends of thread straight .... ..... .. ..... Oaeillatona agardhu
45' Ends of mature threads curved . . . .... ..... . . 46
46 (45') Prominent granules present especially at both ends of each cell . . Oscillatona tenuis
46' Cells without prominent granules .......... . . . . 47
47 (46') Cross walls constricted ................... Oscillatoria chalybea
47' Cross walls not constricted . ....... ..... Oscillatoria formosa
48(43') End of thread long tapering ......................... Oscillatoria splendida
48' End of thread not tapering . .................. Oscillatoria amphibia
49 (24') Cells separate from one another and enclosed in a tube (Cymbella) ...... 251'
49' Cells attached to one another aa a thread or ribbon . ......... . . . 50
50 (49') Cells separating readily into discs or short cylinders, their circular (ace 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 .............. 53
52' Numerous markings in the cell walJ absent (Scenedesmus) ...... ....... 128
53 (52) Wall markings of two types, one coarse, one fine ..... ........... 185
53' Wall markings all fine (Fragilana) ............... . . 54
54(53') Cells attached at middle portion only . ......... Fragilaria crotonensis
54' Cells attached along entire length ...................... 55
55(54') Cell length 2 5-100,, ...... .................. FraRilana capucina
55' Cell length 7 -25,, ..... '. . ....... Fragilaria construens
56(51') Plastid in the form of a spiral band (Spirogyra) .. ............ 57
56' Plastid not a spiral band .....................
CO
57(56) One plastid per cell . ... ...... .... .
57' Two or more plastids per cell ......................
58(57) Threads 18-26, wide ............................. SpiroHyra communi.
58' Threads 28-50(i wide ..........................
59(58') Threads 28-40p wide ............. SpiroByra variajs
59' Threads 40-50M wide . . . . ............ SpirofiYra port.calis
60 (57'» Threads 30-45p wide: 3-4 plastids per cell ........................ SpiroRYra fluviatiha
60' Threads 50-80» wide, 5-8 plastids per cell . . Spirofiyra maiuscula
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 ...... Desrmdium Rrevillii
63' Each cell with a ring of granules near one end Hyalotheca mucosa
64(62') Cells dense green, each plastid reaching to the wall . . ZyBnema
64' Cells light green, plastids not completely filling the cell .
65(64') Width of thread 26-32,,, maximum cell length 6<> . . . ... Zyfinema insifine
65' Width of thread 30-36p, maximum cell length 120p ....... ZyRnema pectmatum
66 (61') Plastid a wide ribbon, passing through the cell axis (Mougeotia). ... . 67
66' Plastid or plastids close to the cell wall (parietal) ..... ... . 69
VII 2-3
-------
67(66) Threads with occasional "knee-joint" bends ............... Mouge^tia Renuflexa
67 ' Threads straight ............................................ 68
68 (67') Threads 19-24M wide, pyrenoids 4-16 per cell ........... Moufjeotia sphaerocarpa
68' Threads 20-34V wide; pyrenoids 4-10 per cell ............ Mouggotia scalaris
69 (661) Occasional cells with one to several transverse wall lines near one end (Oedogonium) 70
69' Occasional terminal transverse wall lines not present ............ 73
70 (69) Thread diameter less than 24ji ................ 7*
70' Thread diameter 25(i or more ......... ....
71 (70) Thread diameter 9-14,. . . ... . - Oedopjonium suecicum
71' Thread diameter 14-23M ....... . Oedogonium boscii
72(70) Dwarf male plants attached to normal thread, when reproducing Oedoeonium idioandrosporum
72' No dwarf male plants produced .......... Oedogoniurn ^rande
73 (69 ') Cells with one plastid which has a smooth surface ........ 7<|
73" Cells with several plastids or with one nodular plastid ..... 78
74(73) Cells with rounded ends . . Stichococcus bacillaris
74' Cells with flat ends (Ulothrix) .............
75(74') Threads 10M or less in diameter .........
75' Threads more than 10|i in diameter ....
76(75) Threads 5-6, in diameter ..... - . . Ulothj-jx variabihs
76'
Threads 6-10H m diameter . Ulothrix tenemma
77(75') Threads 11-17,, in diameter . Ulothrix aequahs
77'
Threads 20-60,, in diameter . Ulothrix Zonata
78 (73') Iodine test for starch positive, one nodular plastid per cell .... 79
78' Iodine test for starch negative, several plastids per cell 80
79 (78) Thread when broken, forming "H" shape segments Microspora amoena
79' Thread when fragrrfented, separating irregularly or between cells (Rhizoclonmm) 100
80 (781) Side walls of cells straight, not bulging A pattern of fine lines or dots present in the wall
but often indistinct (Melosira) ... . 81
80' Side walls of cells slightly bulging Pattern of wall markings not present (Tribonema) 83
81(80) Spine-like teeth at margin of end walls . . . . 82
81' No spine-like teeth present . . .Melosira varians
82 (81) Wall with fine granules, arranged obliquely . Melosira crenulata
82' Wall with coarse granules, arranged parallel to sides Melosira granulata
83 (80') Plastids 2-4 per cell . Tribonema minus
83' Plastids more than 4 per cell . .. . Tribonema bombycinum
84 (231) Plastids present; branching "true" . . . 85
84' Plastids absent, branching "false" . Plectonema tomasmiana
85(84) Branches reconnected, forming a net ... . Hydrodictyon reticulatum
85' Branches not forming a distinct net . ... .86
86 (85') Each cell in a conical sheath open at the broad end (Dinobryon) . .87
86' No conical sheath around each cell. . .... .90
87(86) Branches diverging, often almost at a right angle . ... Dinobryon divergens
87' Branches compace often almost parallel . . . ... 88
88(87') Narrow end of sheath sharp pointed . . - ..89
88' Narrow end of sheath blunt pointed ... . . . .Dinobryon sertulana
VII 2-4
-------
89 (88) Narrow end drawn out into a stalk ... . Dinobryon stipitatum
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 flexilis
91' Short branches on the main thread rebranched two to four times Nitella gracilis
92 <90') Terminal cell eacli with a colorless spine having an abruptly swollen base (Bulbochaete) 93
92' No terminal spines with abruptly swollen bases . . 94
93 Vegetative cells 20-48»* long . Bulbochaete mirabilis
93' Vegetative cells 48-88H long . . .Bulbochaete insignia
94(92') Cells red, brown, or violet .......... . . . Audouinella violacea
94' Cells green . . 95
95 (94') Threads enclosed in a gelatinous bead or mass 96
95" Threads 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 mam thread to branches (Chaetophora) 98
97 (96) Branches (from the main thread) with a central, mam axis Draparnaldia plumosa
97' Branches diverging and with no central main axis Draparnaldia glomerate
98(96') 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
991 Most of the cells essentially alike in density 10°
100(99') Branches few in number, and short, colorless ....... Rhizoclonium hieroglyphicum
100' Branches numerous and green ...
101 (1001) Terminal attenuation gradual, involving two or more cells (Stieeoclonium) 102
101' Terminal attenuation absent or abrupt, involving only one cell (Cladophora) 104
102(101) Branches frequently in pairs ...... 103
102'
Branches mostly single Stigeoclomum stagnate
103(102) Cells in main thread 1-2 times as long as wide StiReoclomum lubricum
103' Cells in main thread 2-3 times as long as wide . Stigeoclomum tenue
104(101') Branching often appearing forked, or in threes Cladophora aegagropila
104' Branches distinctly lateral
105 (104') Branches forming acute angle with main thread, thus forming clusters Cladophora glomerata
105' Branches forming wide angles with the main thread . . lob
106(105') Threads crooked and bent ... Cladophora frac^
106' Threads straight .... ....
107(106') Branches few. seldom rebranching .... Cladophora insignia
107' Branches numerous, often rebranching. Cladophora crispata
108 (41) Plant or tube with a tight surface layer of cells and with regularly spaced swellings (nodes)
1 . . 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 Relatinosa
109' Cells not as loosely arranged spheres
110(109') Plants branch . V.1
110' Plants not branched Schizomens leibleinu
111(110) Clustered branching 11Z
111' B ranches single 115
VII 2-5
-------
112(111) Thraada embedded in gelatinous matrix (Batrachosoermum) 113
112' No gelantinous matrix (Chara) "4
113 (112) Nodal masses of branches touching one another Batraehospermiim vagum
113' Nodal masses of branches separated by a narrow space. . . Batrachospermum moniliforme
114 (1121) Short branches with 2 naked cells at the tip Chara globularis
114' Short branches with 3-4 naked cells at the tip Chara vulgans
115(111') Heterocysts present, plastids absent Stigonema minutum
115' Heterocysts absent, plastids present . Compnopogon coeruleus
116(3') Red eye spot and two flagella present for each cell 125
116' No eye spots nor flagella present II7
117(116') Round to oval cells, held together by a flat gelatinous matrix (Agmenellum) . . ..131
117' Cells not round and not enclosed in a gelatinous matrix 118
118(117') Cells regularly arranged to an unattached disc. Number of cells 2, 4, 8, 16, 32, 64, or
128 133'
118' Cells numerous, membrane attached on one surface 119
119(118') Long hairs extending from upper surface of cells . Chaetopeltis megalocystis
119' No hairs extending from cell surfaces Hildenbrandia rivularis
120 (21) Constriction at the base of every branch Dichotomosiphon tuberosus
120' No constrictions present in the tube (Vaucheria) . . 121
121 (1201) Egg sac attached directly, without a stalk, to the main vegetative tube . .Vaucheria sessilis
121' Egg sac attached to an abrupt, short, side branch 122
122(121') One egg sac per branch .- Vaucheria terrestris
122" Two or more egg sacs per branch Vaucheria geminata
123(1') Cells in colonies generally of a definite form or arrangement 124
123' Cells isolated, in pairs or in loose, irregular aggregates 173
124 (123) Cells with many transverse rows of markings on the wall .185
124' Cells without transverse rows of markings 125
125(124') Cells arranged as a layer one cell thick I26
125' Cell cluster more than one cell thick and not a flat plate . 137
126(125) Red eye spot and two flagella present for each cell Gonium pectorale
126' No red eye spots nor flagella present . . .127
127 (1261) Cells elongate, united side by side in 1 or 2 rows (Scenedesmus) . 128
127' Cells about as long as wide .... ... . 131
128 (127) Middle cells without spines but with pointed ends Scenedesmus dimorphus
128' Middle cells with rounded ends 1*9
129(128") Terminal cells with spines . . ... .130
129' Terminal cells without spines . . . Scenedesmus bnuga
130 (129) Terminal cells with two spines each Scenedesmus quadricauda
130' Terminal cells with three or more spines each . . . Scenedesmus abundans
131 (117) Cells in regular rows, immersed in colorless matrix (Agmenellum ouadriduphcatum) 132
131' Cells not immersed in colorless matrix . - ... .133
132(131) Cell diameter 1 3 to 2 2|» . . . . Agmenellum quadriduplicatum . tenulBaima type
132' Cell diameter 3-5|i . Agmenellum quadriduphcatum. glauca type
133 (1311) Cells without spines, projections, or incisions . . Crucigema quadrata
133' Cells with spines, projections, or incisions . . 134
VII 2-6
-------
134(133') Cells rounded . . Micractinium pusillum
134' Cells angular (Pediastrum) .... .135
135(134') Numerous spaces between cells Pediastrum duplex
135' Cells fitted tightly together . 136
136 (135') Cell incisions deep and narrow . Pediastrum tetras
136' Cell incisions shallow and wide . Pediastrum boryanum
137(125') Cells sharp-pointed at both ends, often arcuate .. . . 138
137' Cells not sharp-pointed at both ends; not arcuate . . 141
138 (137) Cells embedded in a gelatinous matrix ... . Kirchnenella lunar is
138' Cells not embedded in a gelatinous matrix . 139
139 (1381) Cells all arcuate; arranged back to back . Selenastrum gracile
139' Cells straight or bent in various ways, loosely arranged or twisted together
(Ankistrodesmus) . 140
140(139') Cells bent Ankistrodesmus falcatus
140' Cells straight . ... Ankistrodesmus falcatus var acicularis
141 (1371) Flagella present, eye spots often present . 142
141' No flag el la nor eye spots present . 152
142 (141) Each cell in a conical sheath open at the wide end (Dinobryon) 86
142' Individual cells not in conical sheaths . 143
143 (142') Each cell with 1-2 long straight rods extending . Chrysosphaerella longispina
143' No long straight rods extending from the cells. 144
144 (1431) Cells touching one another in a dense colony .... . . 145
144' Cells embedded separately in a colorless matrix . 149
145 (144) Cells arranged radially, facing outward ... . 146
145' Cells all facing in one direction ... . .... . . 147
146(145) Plastida brown, eye spot absent . Synura uvella
146' Plastids green, eye spot present in each cell Pandorina morum
147(145') Each cell with 4 flagella . Spondylomorum quaternarium
147' Each cell with 2 flagella (Pyrobotrys) .148
148(147') Eye spot in the wider (anterior) end of the cell ... ... . Pyrobotrys stellata
148' Eye spot in the narrower (posterior) end of the cell . . . . Pyrobotrys gracilia
149(1441) Plastids brown . Uroglenopsis americana
149' Plastids green . . . . . . . . 150
150(149') Cells 16, 32, or 64 per colony Eudorma elegans
150' Cells more than 100 per colony . . 151
151(150') Colony spherical, each cell with an eye spot. .. . Volvox aureus
151' Colony tubular or irregular: no eye spots (Tetraspora) . 109
15Z (1411) Elongate cells, attached together at one end, arranged radially (Actmastrum) 153
152' Cells not elongate, often spherical 154
153(152) Cells cyhndric . .Actinastrum gracillimum
153' Cells distinctly bulging Actinastrum hantzschii
154(152') Plastids present J55
154' Plastids absent, pigment throughout each protoplast . . . 168
155 (154) Colonies, including the outer matrix, orange to red-brown . . Botryococcus braunii
155' Matrix, if any, not bright colored, cell plastids green 156
VII 2-7
-------
156 (155') Colonies round to oval j^g
156' Colonies not round, often irregular in form 157
157(156') Straight (flat) walls between adjacent cells (Phytocoms) . .. Z78
157' Walls between neighboring cells rounded 15g
158(157') Cells arranged as a surface layer in a large gelatinous tube (Tetraspora) . . 109
158' Colony not a tube, cells in irregular pattern 159
159 (1581) Large cells more than twice the diameter of the small cells (Chlorococcum) . .280'
159' Large cells not more than twice the diameter of the small cells (Palmella) 281
160(156) Cells touching one another, tightly grouped . ... Coe last rum microporum
160' Cells loosely grouped _ , , _ 161
161 (1601) Colorless threads extend from center of colony to cells .... . . 162
161' No colorless threads attached to cells in colony . . 164
162 (161) Cells rounded or straight, oval (Dictyosphaerium) 163
162' Cells elongate, some cells curved Dimorphococcus lunatus
163(162) Cells rounded Dictyosphaerium pulchellum
163' Cells straight, oval Dictyosphaerium ehrenbergianum
164(161') Cells rounded 165
164' Cells oval Oocystis borgei
165 (164) One plastid per cell 166
165' Two to four plastids per cell Gloeococcus schroeteri
166 (165) Outer matrix divided into layers (Cloeocystis) ... . . . . . ... 167
166' Outer matrix homogeneous Sphaerocystts schroeteri
167(166) Colonies angular . .. Gloeocystis planctonica
167' Colonies rounded Gloeocystis gigaa
168(154') Cells equidistant from center of colony (Gomphoaphaeria) . . . . . .. 169
168' Cells irregularly distributed in the colony ...... . . 172
169(168) Cells with pseudovacuoles Gomphospaeria wichurae
169" Cells without pseudovacuoles .... . ... . .. 170
170(169') Cells 2-4|>in diameter (Comphosphaena lacustris) . . 171
170' Cells ovate . . Gomphosphaeria aponina
171(170) Cells spherical. . Gomphosphaeria lacustris, kuetzmgianum type
17'' Cells 4-15 in diameter . . . ... Gomphosphaeria lacustris, colhnaii type
172 (1681) Cells ovid, division plane perpendicular to long axis (Coccochloris) . 286
172' Cells rounded, or division plane perpendicular to short axis (Anacystis) . . 286'
173 (1231) Cells with an abrupt median transverse groove or incision . . - .174
173' Cells without an abrupt transverse median groove or incision . ... .184
174 (173) Cells brown, flagclla present (armored flagellates) . . . 175
174' Cells green, no flagella (desrmds) . 178
175(174) Cell with 3 or more long horns Ceratium hirundinella
175' Cell without more than 2 horns 176
176 (1751) Cell wall of very thin smooth plates Glenodinium palustre
176' Cell wall of very thick rough plates (Peridimum) 177
177 (1761) Ends of cell pointed Peridimum wisconsinense
177' Ends of cell rounded Peridimum cinctum
178 (174') Margin of cell with sharp pointed , deeply cut lobes or long spikes . . 179
178' Lobes, if present, with rounded ends 182
VII 2-8
-------
179 (178) Median incision narrow, linear Micrasteriaa truncata
179' Median incision wide. "V" or "U" shaped (Staurastrum) 180
180 (179) Margin of cell with long spikes Staurastrum paradoxum
ISO1 Margin of cell without long spikes 181
181 (180") Ends of lobes with short spines Staurastrum polymorphum
181' Ends of lobes without spines Staurastrum punctulatum
182 (1781) Length of cell about double the width , Euastrum oblongum
182' Length of cell one to one and one-half times the width (Coamarium) ... . ... 183
183 (1821) Median incision narrow linear Coamarium botrytia
183' Median incision wide, "U" shaped Cosmarium portianum
184(173') Cells triangular Tetraedron muticum
184' Cells not triangular . 185
185 (124) Cells with one end distinctly different from the other ... 186
185' Cells with both ends essentially alike 225
186 (185) Numerous transverse (not spiral) regularly spaced wall markings present (diatoms) 187
186' No transverse regularly spaced wall markings 193
187 (186) Cells curved (bent) in girdle view Rhoicosphenia curvata
187' Cells not curved in girdle view 188
188 (187') Cells with both fine and coarse transverse lines Meridion circulare
188' Cells with transverse lines all alike in thickness 189
189 (1881) Cells essentially linear to rectangular; one terminal swelling larger than the other
(Asterionella) 190
189' Cells wedge-shaped, margins sometimes wavy (Gomphonema) 191
190 (189) Larger terminal swelling 1-1/2 to 2 times wider than the other Asterionella formosa
190' Larger terminal swelling less than 1-1/2 times wider than the other. .Asterionella Rracilhma
191 (1891) Narrow end enlarged in valve view Gomphonema geminatum
191' Narrow end not enlarged in valve view 192
192 (1911) Tip of broad end about as wide as tip of narrow end in valve view . Gomphonema parvulum
192' Tip of broad end much wider than tip of .narrow end in valve view. Comphonerna olivaceum
193 (1861) Spine present at each end of cell Schroederia aetigera
193' No spine on both ends of cell 194
194 (1931) Pigments in one or more plastids 195
194' No plastid; pigments throughout the protoplast Entophysalis lemamae
195 (194) Cells in a conical sheath (Dinobryon) 86
195' Cells not in a conical sheath .196
196 (195') Cell covered with scales and long spines MaHomenas caudata
196' Cells not covered with scales and long spines 197
197(196') Protoplasts separated by a space from a rigid sheath (lorica) 198
197' No loose sheath around the cells 202
.199
199 (198') Lorica opaque; yellow to reddish or brown Trachelomonas crebea
199' Lorica transparent; colorless to brownish (Chryaococcusl . .. . .. . .200
200(199')' Outer membrane (lorica) oval Chrysococcus ovahs
200' Outer membrane (lorica) rounded . 201
vn 2-9
-------
201(200') Lorlca thickened around opening Chry.ococcus rutescens
maior
201" Lorica not thickened around opening Chrysococcus
202 (197 ') Front end flattened diagonally ............................................. 203
203' Front end not flattened diagonally ............................ .......... .... 206
203(202) Plaatids bright blue -green (Chroomonas) ............................... 204
203' Plastids brown, red, olive-green, or yellowish ............................ .. 205
204 (203) Cell pointed at one end ....................................... Chroomonas nordstetu
204' Cell not pointed at one end ................................ Chroomonas setoniensis
205(203') Gullet present, furrow absent ..... ........................ Cryptomonae erosa
205' Furrow present, gullet absent ................................. Rhodomonaa
206(202') Plastids yellow-brown ......................................... Chromulina roaanoffi
206' Plastids not yellow-brown; generally green . .......................... 207
207(206') One plastid per cell .................................................. 208
207' Two to several plastids per cell ................................ ' ' 211
208(207) Cells tapering at each end ...................... Chlorogonlum euchlorum
208" Cells rounded to oval ................................................ ~TT. - 209
209 (208') Two flagella per cell (Chlamydomonas) ................... 210
209' Four flagella per cell ............................ " . ." Cater'ia multifil'is
210(209) Pyrenoid angular, eye spot in front third of cell ........ Chlamydomonas reinhardi
210 Pyrenoid circular, eye spot in middle third of cell ........... . Chlarnvdomon"as eloboaa
211(207.) Two plastids per cell ......................... Cryptoglena pjgra
211' Several plastids per cell ................................ -- ^212
212 (2111) Cell compressed (flattened) (Phacus) .... . 213
212' Cell not compressed ......................... ' ' ,..
213(212) Posterior spine short, bent .................. Phacus pleuronectee
213 Posterior spine long, straight ........... . . . . Phacus longicluda"
214 (212) Cell margin rigid ................................... 215
2141 Cell margin flexible (Euglena) ... .... 217
215(214) Cell margin with spiral ridges ............................... Phacus pyrum
^15 Cell margin without ridges, but may have spiral lines (Lepocinclis) . . . . . . 216
216 (215') Posterior end with an abrupt, spine-like tip .............. . Lepocinclis ovum
216' Posterior end rounded ..................... Lepocinclis
217(214') Green plastids hidden by a red pigment in the cell . . . . Euglena sanguinea
217' No red pigment except for the eye spot ....... . . . ~~ - 218
218(217') Plastids at least 1/4 the length of the cell ............. .......... 219
218' Plastids discoid or at least shorter than 1/4 the length of the cell . 220
219 (218) Plastids two per cell ............................. ... Euelena agll,8
219' Plastids several per cell, often extending radiately from the center. . Euglena viridis
220(218') Posterior end extending as an abrupt colorless spine .............. 221
220' Posterior end rounded or at least with no colorless spine .......... 222
221(220) Spiral markings very prominent and granular ..................... Euglena spirogyra
221' Spiral markings fairly prominent, not granular ..................... Euglena oxyuris
222 (220') Small, length 35-55, ........................... Euglena gracilis
222' Medium to large, length 65, or more ............. . ...... - 223
223(222') Medium in size; length 65-200, .......................... 224
223- Large in size, length 250-290M ................ ".'.'.'. '.'. . '.".'..'. .Eugiena e'hren'bergii
vn 2-10
-------
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 . . Euglena deses
225 (1851) Cells distinctly bent (arcuate); with a spine or nan owing to a point at both ends 226
225' Cells not arcuate - 23°
226 (225) Vacuole with particles showing Brownian movement at each end of cell Cells not in
clusters (Closterium) 2-7
226' No terminal vacuoles. Cells may be in clusters or colonies . 228
2Z7 (226) Cell wide, width 30-70|i . . . . Closterium momhferum
227' Cell long and narrow, width up to 5M Closterium aciculare
228 (2261) Cell with a narrow abrupt spine at each blunt end Ophiocytium capitatum
2Z81 No blunt ended cells with abrupt terminal spines . 229
229 (228') Sharp pointed ends as separate colorless spines . 193
2Z91 Sharp pointed ends as part of the green protoplast 137
230 (225) One long spine at each end of cell ... . . 231
230' No long terminal spines . . 232
231 (230) Cell gradually narrowed to the spine . -137
231' Cell abruptly narrowed to the spine Rhizosolema gracihs
232 A regular pattern of fine lines or dots in the wall (diatoms) 233
232' No regular pattern of fine lines or dots in the wall . . 276
233 (50, Cells circular in one (valve) view, short rectangular or square in other (girdle) view . 234
232)
233' Cells not circular in one view. -*°
234 (233) Valve surface with an inner and outer (marginal) pattern of striae (Cyclotella) 235
234' Valve surface with one continuous pattern of striae (Stephanodiscus) . 238
235(234) Cells small. 4-10,» in diameter Cyclotella filomcrata
235' Cells medium to large, 10-80 in diameter . . . 236
236 (2351) Outer half of valve with two types of lines, one long, one short 23?
236' Outer half of valve with radial lines all alike Cyclotella meneghiniana
237 (236) Outer valve zone constituting more than 1/2 the diameter Cyclotella bodanica
237' Outer valve zone constituting more than 1/2 the diameter. Cyclotella compta
21<)
238(234') Cell 4-25|i in diameter. . _,
238' Cell25-65M in diameter . .. Stephanodiscus mafiarae
239(238) Cell with two transverse bands, in girdle view Steohanodiseus binderanus
239' Cell without two transverse bands, in girdle view .. Stephanodiscus hantzschii
240(233') Cells flat, oval (Cocconeis) *
240' Cells neither flat nor oval
241(240) Wall markings (striae) 18-20 in 10M Cocconeis pediculus
241' Wall markings (striae) 23-25 in 10M. Cocconeis placentula
243
242(240') Cell aigmoid in one view
242' Cell not sigmoid in either round or point ended (valve) or square ended (girdle) surface
view.
243(242) Cell sigmoid in valve surface view . CYro3iBma attenuatum
243' Cell sigmoid in square ended (girdle) surface view . . ... Nitzschia aciculans
244 (242') Cell longitudinally unsymmetrical in at least one view . 24j>
244' Cell longitudinally symmetrical
245(244) Cell wall with both fine and coarse transverse lines (striae and costae) . 246
245' Cell wall with fine transverse lines (striae) only . - 247
VII 2-11
-------
246 (245)
246'
247 (245)
247'
248 (247)
248'
249 (248')
249'
250(247')
250'
251 (250)
251'
252 (2511)
(246)
252'
253 (252')
253'
254 (244')
254'
255 (254)
255'
Z56 (Z551)
256'
2S7 (254)
257'
258 (257)
258'
259 (257')
259'
260 (Z591)
260'
2ol (260)
261'
262 (261)
262'
i
-------
267 (2661) Striae distinctly composed of clots (punctae)
267' Striae essentially as continuous lines
268 (2671) Central clear area on valve face rectangular
268' Central clear area on valve face oval
269 (268') Cell length 29-40)1. ends slightly capitate
269' Cell length 30-l20(i, ends not capitate
270 (260') Knob at one end larger than at the other (Asterionella)
270' Terminal knobs if present equal in size (Synedra)
Navicula lann-olata
Navicula graciloidcs
Navicula eryptoccphala
Navicula racliosa
189
271
271 (2701)
271'
272 (2711)
272'
273 (272')
273'
274
274'
275 (274')
275'
276 (232')
276'
277 (276)
277'
278 (277')
278'
279 (278')
279'
280 (2791)
Z80'
281 (159')
281'
282 (281')
282'
Clear space (pseudonodule) in central area
No pseudonoriule in central area
Sides parallel in valve view, each end with an enlarged nodule
Sides converging to the ends in valve view
Valve linear to lanceolate-linear, 8-12 striae per 10p
Valve narrowly linear-lanceolate, 12-18 striae per 10(1
Valve 5-6|» wide ....
Valve 2-4M wide ... . . .
Synrdra pulchclla
272
Synedra ulna
.274
275
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 augustissima
Green to brow.i pigment in one or mure plastids
No plastids, blue and green pigments throughout protoplast
Cells long and narrow or flat
Cells rounded
Straight, flat wall between adjacent cells in colonies
Rounded wall between adjacent cells in colonies
277
284
233
278
fhytoconis botryoides
279
Cell either with 2 opposite wall knobs or colony of 2-4 cells surrounded by distinct mem-
brane or both .... l64
Cell witnout 2 wall knobs, colony not of 2-4 cells surrounded by distinct? 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 Hurmcola
Palmclla murosa
282
283
Chlorella rllipsoidoa
283 (282)
283'
284 (276')
284'
285 (25)
285'
Cell 5-10^1 in diameter, pyrenoid indistinct
Cell 3-5u in diameter, pyrenoid distinct
Cell a spiral rod . . .
Cell not a spiral rod
Thread septate (with crosswalls)
Thread non-septate (without crosswalls)
Chlorella vulj-aris
Chlorella pyrpnoidosa
285
286
Arthrospira jenncri
Spirulma nordstrdtii
286 (172) Cells dividing in a plane at right angles to the long axis
(284')
286' (1721) Cells sperical or dividing in a plane parallel to the long axis (Anacystis)
287 (286') Cell containing pseudovacuoles
287' Cell not containing pseudovacuoles.
Coccochluria stagnina
287
Anacystis cyanea
288
VII 2-13
-------
288(287') Cell 2-6B in diameter; aheath often colored Anacyatis montana
288' Cell 6-50n in diameter; sheath colorless 289
289 (2881) Cell 6-12n in diameter; cells in colonies are mostly spherical . . . . Anacystis thermalis
289' Cell 12-50(1 in diameter; cells in colonies are often angular. . Anacyatis dimidiata
VII 2-14
-------
APPENDIX
CLASSIFICATION-FINDER FOR NAMES OF
AQUATIC ORGANISMS IN WATER SUPPLIES
AND POLLUTED WATERS
-------
FOREWORD
The following work is more easily defined in terms of what it is not,
than what it is; it is not a "key" in the usual biological sense of the
word, nor is it a glossary of a dictionary. It is rather a device for
determining what general kind of organism or group is designated by
some unrecognized name, be it common or scientific.
If one has access to one or more of the references cited, he can find
the same name, and learn much more about it; but not everyone has
all of these books, and the information is often couched in highly
technical terms.
The nonbiologist would be well advised to read Part I before attemp-
ting to use the Finder. The experienced biologist on the other hand
may proceed directly to the index and quickly be referred to the larger
group to which his unknown organism belongs.
No professional systematist will find himself completely at home. In
an effort to present a relatively simple concept of relationships
couched in standard terms for all groups or organisms, some violence
was done to certain highly sophisticated systems of classification. It
is hoped, however, that the layman will find accuracy sufficient for his
needs, and the specialist will be referred to technical literature where
he can satisfy his needs for greater detail.
While every effort has been made to ensure accuracy, it is inevitable
that errors have crept in. Please call them to our attention.
Grateful appreciation is extended to Michael E. Bender and Charles L.
Brown, Jr., both former Biologists with the Water Pollution Training
Activities, for their valuable contributions and encouragement.
H.W. Jackson
Chief Biologist
National Training Center
-------
PART I. The System of Biological Classification
PART II. Outline of Biological Classification
-------
PART I
The System of
Classification
-------
CLASSIFICATION - FINDER
for
NAMES OF AQUATIC ORGANISMS
in
WATER SUPPLIES AND POLLUTED WATERS
Part I. The System of Classification
I INTRODUCTION
A Every type of living creature
has a favorite place to live.
There are few major groups that
are either exclusively terres-
trial or aquatic. The following
remarks will therefore apply in
large measure to both, but pri-
mary attention will be directed
to aquatic types.
B One of the first questions usu-
ally posed about an organism is:
"What is it?", usually meaning
"What is it's name?". The nam-
ing or classification of bio-
logical organisms is a science
in itself (taxonomy). Some of
the principles involved need to
be understood by anyone working
with organisms however.
1 Names are the "key number",
"code designation", or "file
references" which we must
have to find information
about an unknown organism.
2 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 subdivi-
sions. There are over a
million and a half items (or
species) included in the
system of biological nomen-
clature (very few libraries
have a million books).
3 The system of biological no-
menclature is regulated by
international congresses.
a It is based on a system of
groups and super groups,
of which the foundation
(which actually exists in
nature) is the species.
Everything else has been
devised by man and is sub-
ject to change and revision
as man's knowledge and
understanding increase.
b The basic categories em-
ployed are as follows:
(1) Similar species are
grouped into genera
(genus)
(2) Similar genera are
grouped into families
(3) Similar familes are
grouped into orders
(4) Similar orders are
grouped into classes
(5) Similar classes are
grouped into phyla
(phylum)
(6) Similar phyla are
grouped into kingdoms
The scientific name of an or-
ganism is its genus name plus
its species name. This is ana-
logous to our system of sur-
names (family names) and given
names (Christian names).
a The generic (genus) name
is always capitalized and
the species name written
with a small letter. They
should also be underlined
or printed in italics when
used in a technical sense.
For example:
Homo sapiens - modern man
Homo neanderthalis -
neanderthal man
Esox niger - Chain pickerel
Esox lucius - northern pike
Esox masquinongy -
muskellunge
b 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
BI.AQ. 24. 10.66
-------
RELATIONSHIPS BE1WEEN LIVING ORGANISMS
PLANTS 5
ORGANIC MATERIAL
PRODUCED, USUALLY
BY PHOTOSYNTHESIS
ENERGY FLOWS FROM LEFT TO RIGHT, GENERAL EVOLUTIONARY
SEQUENCE IS UPWARD
ANIMALS 81
ORGANIC MATERIAL INGESTED OR
CONSUMED
DIGESTED INTERNALLY
ENERGY STORED
ENERGY RELEASED
FUNGI 250
ORGANIC MATERIAL
REDUCED
BY EXTRACELLULAR
DIGESTION AND IN-
TRACELLULAR META-
BOLISM TO MINERAL
CONDITION
ENERGY RELEASED
FLOWERING PLANTS
AND GYMNOSPERMS 76
CLUB MOSSES, FERNS 76
LIVERWORTS, MOSSES 73
ALGAE 12
ARACHNIDS 167
INSECTS 154
CRUSTACEANS 129
SEGMENTED WORMS 121
MOLLUSCS 172
MOSS ANIMALS 120,181
WHEEL ANIMALS 116
ROUNDWORMS 113
FLATWORMS 108
MAMMALS 2 3
BIRDS 242
REPTILES 241
AMPHIBIANS 240
FISHES 195
PROCHORDATES 191
STARFISH GROUPS 185
BASIDIOMYCETES 266
ASCOMYCETES 265
HIGHER PHYCOMYCETES
261
JELLYFISH - CORAL GROUP 103
SPONGES 99
DEVELOPMENTOFMULTICELLULARORCOENOCYTICORGANISMS
DIATOMS 38
PIGMENTED FLAGELLATES
HIGHER PROTISTA
PROTOZOA 82
AMOEBOID PROTOZOA 86 CILLIATED PROTOZOA 92
FLAGELLATED PROTOZOA 85 SPOROZOA 98
12 I (COLORLESS FLAGELLATES 85 SUCTORIA 97
LOWER PHYCOMYCETES
261
DEVELOPMENT OF A NUCLEAR MEMBRANE
BLUE GREEN ALGAE 7
PHOTOTROPIC
BACTERIA 252
CHEMOTROPIC BACTERIA
252
LOWER PROTISTA (ORMONERA)
NOTE: NUMERALS REFER TO PARAGRAPHS IN PARTS 2 AND 3.
W. B. COOKE AND H. W. JACKSON, AFTER WHITTAKER
ACTINOMYCETES 253
SPIROCHAETES 255
MYXOBACTERIA 254
PARASITIC
BACTERIA 251
AND VIRUSES
SAPROBIC BACTERIA 251
BI.ECO.pl.2b.4.66
-------
Classification - Finder
genus Anabaena (an alga),
we must simply use the
generic name, and:
Anabaena^ planctonica,
Anabaena gonstricta, and
Anabaena flos-aquae
are three distinct species
which have different signi-
ficances to water treatment
plant operations.
A complete list of the various
categories to which an organism
belongs is known as its "classi-
fication". For example, the
classification of a type of
frog spittle, a common fila-
mentous alga, and a crayfish
or crawdad are shown side by
side below. Their scientific
names are Spirogyra crassii and
Cambarus sciotensis.
a Examples of the classifica-
tion of an animal and a
plant:
(Frog Spittle)
Plantae
Chlorophyta
Chlorophyceae
Zygnematales
Zygnemataceae
Spirogyra
crassa
Kingdom
Phylum
Class
Order
Family
Genus
Species
(Crayfish)
Animalia
Arthropoda
Crustacea
Decapoda
Palaemonidae
Cambarus
sciotensis
These seven basic levels of
organization are often not
enough for the complete de-
signation of one species
among thousands; however,
and so additional echelons
of terms are provided by
grouping the various cate-
gories into "super..."
groups and subdividing them
into "sub..." groups as:
Superorder, Order, Suborder,
etc. Still other category
names such as "tribe", "di-
vision", "variety", "race",
"section", etc. are used on
occasion.
c Additional accuracy is gained
by citing the name of the
authority who first described
a species (and the date) im-
mediately following the spe-
cies name. Authors are also
often cited for genera or
other groups.
d A more complete classification
of the above crayfish is as
follows:
Kingdom Animalia
Phylum Arthropoda
Class Crustacea
Subclass Malacostraca
Order Decapoda
Section Nephropsidea
Family Astacidae
Subfamily Cambarinae
Genus Cambarus
Species sciotensis Rhoades
1944
e It should be emphasized that
since all categories above
the species level are essen-
tially human concepts,! there
is often divergence of opin-
ion in regard to how certain
organisms should be grouped.
Changes result as knowledge
grows.
f The most appropriate or cor-
rect name for a given species
is also sometimes disputed,
and so species names too are
changed. The species itself,
as an entity in nature, how-
ever, is relatively timeless
and so does not change to
man's eye.
II THE GENERAL RELATIONSHIPS OF
LIVING ORGANISMS
:
A Living organisms (as contrasted to
fossil types) have long been group-
ed into two kingdomfi: Plant King-
doms and Animal Kingdoms. Modern
developments however have made this
-------
Classification - Finder
simple pattern technically unten-
able. 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;
1 Plantae: photosynthetic;
synthesizing their own organic
substance from inorganic min-
erals. 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 metabolized and reduced to
the mineral condition. Ecolo-
gically known as REDUCERS.
C Each of these groups includes
simple, single celled representa-
tives, persisting at lower levels
on the evolutionary stems of the
higher organisms.
1 These groups span the gaps be-
tween the higher kingdoms with
a multitude of transitional
forms. They are collectively
called PROTISTA.
2 Within the protista, two prin-
ciple sub-groups can be defined
on the basis of relative com-
plexity of structure:
a The bacteria and blue algae,
lacking a nuclear membrane,
may be considered as the
lower protista or MONERA.
b The single celled algae and
protozoa having a nuclear
membrane, are best referred
to simply as the higher
protista.
-------
PART II
Outline
of Biological
Classification
-------
Classification - Finder
Part II. Biological Classification
I INTRODUCTION
A What is it?
B Policies
C Procedures
II PLANT KINGDOM
A "Algae" defined
1
2
3
4
5
6
B PHYLUM CYANOPHYTA - blue-green 7
algae
CLASS Hyxophyceae 8
Order Chroococcales 9
Order Hormogonales 10
Suborder Hetercystineae 11
C PHYLUM CHLOROPHYTA - green 12
algae
CLASS Chlorophyceae 13
Order Volvocales 14
Order Ultrichales 15
Order Chaetophorales 16
Order Chlorococcales 17
Order Siphonales 18
Order Zygnematales 19
Order Tetrasporales 20
Order Ulvales 21
Order Schizogonales 22
Order Oedogoniales 23
Order Cladophorales 24
CLASS Charophyceae 25
Order Charales 26
D PHYLUM CHRYSOPHYTA - yellow- 27
green algae or yellow-brown
algae
CLASS Xanthophyceae 28
Order Heterocapsales
Order Heterococcales
CLASS Chrysophyceae -
yellow-green algae
Order Chrysomondales
Order Rhizochrysidales
Order Chrysosphaerales
Order Chrysocapsales
Order Chrysotrichales
30
31
32
33
34
35
36
37
Order Rhizochloridales
29
CLASS Bacillariophyceae - 38
Diatoms
Order Pennales - pennate 39
diatoms
Order Centrales - centric 40
diatoms
E PHYLUM EUGLENOPHYTA - eugle- 41
noid algae
F PHYLUM PYRRHOPHYTA - yellow 42
brown algae
CLASS Desmokontae 43
Order Desmomonadales 44
CLASS Dinophyceae - 45
dinoflage Hates
Order Gymnodiniales 46
Order Peridiniales 47
Order Dinocapsales 48
Order Chloromonadales 49
CLASS Cryptophyceae 50
G PHYLUM CHLOROMONADOPHYTA 51
H PHYLUM RHODOPHYTA - red algae 52
CLASS Rhodophyceae 53
Order Bangiales 54
Order Nemalionales 55
Order Gelidiales 56
-------
Classification - Finder
Order Cryptonemiales
Order Gigartinales
Order Rhodymeniales
Order Ceramiales
57
58
59
60
I PHYLUM PHAEOPHYTA - brown algae 61
CLASS Phaeophyceae 62
Order Ectocarpales 63
Order Sphacelariales 64
Order Tilopteridales 65
Order Chordiales 66
Order Desmarestiales 67
Order Punctariales 68
Order Dictyosiphonales 69
Order Laminarlales 70
Order Fucales 71
Order Dictyotales 72
J PHYLUM BRYOPHYTA 73
CLASS Hepaticae - liverworts 74
CLASS Musci - mosses 75
K VASCULAR PLANT GROUP 76
Emergent vegetation 77
Rooted plants - floating leaves 78
Submerged vegetation 79
Free floating plants 80
III ANIMAL KINGDOM 81
A PHYLUM PROTOZOA - protozoa 82
CLASS Mastigophora 83
Subclass phytomastlgina 84
Subclass zoomastigina 85
CLASS Sarcodina - amoeboid 86
protozoa
Order Amoebina
Order Foraminifera
Order Radiolaria
Order Heliozoa
87
88
89
90
Order Mycetozoa (Myxomycetes) 91
CLASS Ciliophora - ciliates 92
Order Holotricha 93
Order Spirotricha 94
Order Peritricha 95
Order Chonotricha 96
CLASS Suctoria - suctoria 97
CLASS Sporozoa 98
B PHYLUM PORIFERA - sponges 99
CLASS Calcispongea 100
CLASS Hyalospongea 101
CLASS Demospongea 102
C PHYLUM COELENTERATA 103
CLASS Hydrozoa - hydroids 104
CLASS Scyphozoa - jellyfish 105
CLASS Actinozoa (Anthozoa) - 106
corals
D PHYLUM CTENOPHORA - comb 107
jellies
E PHYLUM PLATYHELMINTHES - 108
flatworms
CLASS Turbellaria - turbella- 109
rians
CLASS Trematoda - fluke 110
CLASS Cestoldea - tapeworms 111
F PHYLUM NEMERTEA - proboscis 112
worms
G PHYLUM NEMATODA - threadworms,113
roundworms
-------
Classification - Finder
H PHYLUM NEMATOMORPHA -
Horsehair worms
114
I PHYLUM ACANTHOCEPHALA - thorny 115
headed worms
J PHYLUM ROTIFERA - rotifer, 116
wheel animalcules
K PHYLUM GASTROTRICHA - gastro- 117
trichs
L PHYLUM KINORHYNCHIA
M PHYLUM PRIAPULIDA
N PHYLUM ENDOPROCTA
118
119
120
0 PHYLUM ANNELIDA - segmented 121
worms
CLASS Polychaeta - polychaet 122
worms
CLASS Oligochaeta - bristle 123
worms
CLASS Hirudlnea - leeches 124
CLASS Archiannelida 125
CLASS Echiuroidea 126
CLASS Sipunculoidea - peanut 127
worms
P PHYLUM ARTHROPOOA - jointed 128
legged animals
CLASS Crustacea - crustaceans 129
Subclass Branchiopoda 130
Order Anostraca - fairy 131
shrimps
Order Notostraca - tadpole 132
shrimps
Order Conchostraca - clam 133
shrimps
Order Cladocera - water fleas!34
Subclass Ostracoda - seed 135
shrimps, ostracodes
Subclass Copepoda - copepods 136
Subclass Branchiura - fish 137
lice
Subclass Cirripedia -
barnacles
Subclass Malacostraca
Order Leptostraca
138
139
140
Order Hoplocardia 141
(Stomatopoda) - mantis shrimps
Order Syncarida
Order Peracarida
Suborder Mysidacea
Suborder Cumacea
Suborder Tanaidacea
142
143
144
145
146
Suborder Isopoda - sowbugs,147
pillbugs
Suborder Amphipoda - scuds 148
Order Eucarida 149
Suborder Euphausiacea - 150
krill
Suborder Decapoda - shrimp,151
lobster, crab
Macrurous group (4 tribes) 152
shrimps, prawns, lobsters,
crayfish
Brachyurous group 153
(2 tribes) - crabs and hermit
crabs
CLASS Insecta - the Insects 154
Orders represented by imma- 155
ture stages only.
Order Plecoptera - stone- 156
flies
Order Ephemeroptera -
mayflies
157
Order Odonata - dragon and 158
damselflies
Order Megaloptera - alder- 159
flies, dobsonflies, fishflies
Order Neuroptera - spongilla-160
flies
-------
Classification - Finder
Order Trichoptera - caddis- 161
flies
Order Lepidoptera - aquatic 162
caterpillars
Order Dlptera - two winged 163
flies
Orders including aquatic 164
adults
Order Coleoptera - beetles 165
Order Hemiptera - true bugs 166
CLASS Arachnoldea - spiders, 167
scorpions, mites
Order Xlphosoura - horse- 168
shoe or king crabs
Order Hydracarina - aquatic 169
mites
Order Pantopoda (Pycnogonida)-170
pycnogonids
Order Tardigrada 171
Q PHYLUM MOLLUSCA 172
CLASS Amphlneura - chitons 173
CLASS Gasteropoda - snails 174
Order Prosobranchiata 175
Order Opisthobranchiata 176
Order Pulmonata - air breath- 177
ing snails
CLASS Scaphopoda - tusk 173
shells
CLASS Bivalvia 179
(Pelecypoda)
CLASS Cephalopoda - squid, ISO
octipus, nautilus
R PHYLUM BRYOZOA (Ectoprocta) - 181
Moss animals
S PHYLUM BRACHIOPODA - lamp 182
shells
T PHYLUM CHAETOGNATHA - arrow 183
worms
U PHYLUM PHORONIDEA - tufted 184
worms
V PHYLUM ECHINODERMATA -
echinoderms
185
CLASS Asteroidea - starfishes 186
CLASS Ophiuroidea - brittle 187
stars
CLASS Echinoidea - sea urchins 188
CLASS Holothuroidea - sea 189
cucumbers
CLASS Crinoidea - sea lilies 190
W PHYLUM CHORDATA - chordates 191
Subphylum Hemichordata - 192
Acorn worms
Subphylum Urochordata -
tunicates, sea squirts
193
Subphylum Cephalochordata - 194
lancelets
Subphylum Vertebrata 195
(Craniata) - vertebrates
CLASS Agnatha - jawless 196
fishes
Order Myxiniformes - 197
hagfishes
Order Petromyzontiformes - 198
lampreys
CLASS Chrondrichthys -
cartilage fishes
199
Order Squaliformes - sharks 200
Order Rajiformes - skates, 201
rays
Order Chimaeriformes -
chlmaeras
202
CLASS Osteichthys (Pisces) - 203
bony fishes
Order Acipenseriformes - 204
sturgeons
Order Polyodontidae -
paddle fishes
205
-------
Classification - Finder
Order Semionoteformes - gars 206
Order Amiiformes - bowfins 207
Order Clupeiformes - soft 208
rayed fishes
Family Clupeidae - herrings 209
Family Salmonidae - trouts, 210
salmon
Family Esocidae - pikes, 211
pickerels
Family Serranidae - sea 228
basses
Order Myctophiformes -
lizard fishes
Order Cypriniformes -
212
213
Family Cyprinldae - minnows, 214
carps
Family Catostomidae - suckers215
Family Ictaluridae - fresh- 216
water catfishes
Order Anguilliformes - eel- 217
like fishes
Order Notacanthiformes -
spiny eels
218
Order Beloniformes - needle- 219
fishes, flying fishes
Order Cyprinodontiformes - 220
killifishes, livebearers
Order Gadiformes - cods and 221
hakes
Order Gasterosteiformes - 222
stickelbacks
Order Lampridiformes - Opahs, 223
ribbon fishes
Order Beryciformes - beard- 224
fishes
Order Percopsiformes - trout 225
and pirate perches
Order Zeiformes - dory
226
Order Perciformes - spiny- 227
rayed fishes
Family Centrarchidae -
sunfishes, freshwater
basses
229
Family Percidae - perch 230
Family Sciaenidae - drum 231
Family Cottidae - sculpins 232
Family Magilidae - mullets 233
Order Pleuronectiformes - 234
flounders
Order Echeneiformes - remoras235
Order Gobiesociformes - 236
clingfishes
Order Tetraodontiformes - 237
spikefishes
Order Batrachoidiformes - 238
toadfishes
Order Lophiiformes -
goosefishes
239
CLASS Amphibia - frogs, toads,240
salamanders
CLASS Reptilia - turtles, 241
snakes, lizards
CLASS Aves - birds
CLASS Mammalia - whales,
seals, walrusses
IV FUNGUS KINGDOM
A Bacteria
Eubacteria
Actinomycetes
Myxobacteria
Spirochaetes
Other bacterial types
B FUNGI
"Phycomycete" group
242
243
250
251
252
253
254
255
256
260
261
-------
Classification - Finder
CLASS Chytrldiomycetes 262
CLASS Oomycetes 263
CLASS Zygomycetes 264
CLASS Ascomycetes 265
CLASS Basidiomycetes 266
CLASS Fungi Imperfecti 267
10
------- |