A CURRICULUM ACTIVITIES GUIDE TO
WATER
POLLUTION
AMD
ENVIRONMENTAL
VOLUME 2
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A CURRICULUM ACTIVITIES GUIDE
T 0
WATER POLLUTION
and
ENVIRONMENTAL STUDIES:
APPENDICES
U. S. ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF WATER PROGRAMS
MANPOWER DEVELOPMENT STAFF
TRAINING GRANTS BRANCH
1972
For sale by the Superintendent of Documents, U.S. Government Printing Office, Washington, B.C., 20402 - Price $2.25
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This guide was prepared by the Til ton Water Pollution Program, financed
by Grant No. 1TT1-WP-41-01 and supplemental grants from Training Grants
Branch, Office of Water Programs, Environmental Protection Agency and
by a grant from the Ford Foundation. The work of editing and compiling
the guide was done by:
John T. Hershey
Head, Science Department
Germantown Academy
Fort Washington, Pennsylvania
Albert L. Powers
Head, Science Department
Brewster Academy
Wolfeboro, New Hampshire
Stephen P. McLoy
Teacher of Political Theory
Til ton School
Til ton, New Hampshire
Alan D. Sexton
Teacher of Science
George School
Newtown, Pennsylvania
Information on revisions and additionally planned volumes of the guide
may be obtained from:
Training Grants Branch
Office of Water Programs
U. S. Environmental Protection Agency
Washington, D. C. 20460
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TABLE OF CONTENTS
Appendix
1. WATER QUALITY PARAMETERS A-l
A. Chemistry A-l
1. Acid Base Parameters A-l
a. Acidity A-l
b. Alkalinity A-9
c. pH A-13
2. Dissolved Gases A-14
a. C02 A-14
b. Chlorine A-14
c. Dissolved Oxygen A-15
-—,
•-I 3. Dissolved and Suspended Solids A-19
a. Chloride A-19
Q b. Hardness-Calcium, Magnesium, Total A-21
c. Iron A-24
d. Nitrate A-25
r< e. Nitrite A-26
"T f. Phosphate A-26
-* g. Sulfate A-28
h. Turbidity A-30
4. Oxygen Demand A-31
5. Interpretation A-37
6. Bibliography A-44
B. Bacteriology A-45
1. Total Coliform A-45
2. Fecal Coliform A-46
3. Fecal Streptococci A-48
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Appendix
4. General Procedures A-49
a. Sterilization A-49
b. Preparation of Media „ A-50
c. Preparation of Solutions A-53
d. Collection of Water Sample „ A-55
e. Filtration Volumes - Selection and Dilution . „ A-55
f. Preparation of Filter for Incubation „ A-59
g. Incubation „ A-62
h. Counting Techniques „ A-63
i. Disposal of Cultures A-65
5. Bibliography A-65
C. Aquatic Biology „ A-67
1. The Basis of the Biological Evaluation of
Pollution A-67
2. The Identification of Aquatic Organisms . A-68
3. Biological Field Methods . A-96
a. Benthos A-96
b. Periphyton or Aufwuchs A-105
c. Plankton A-105
d. Nekton A-106
e. Sample Data Sheets A-107
4. Biological Laboratory Methods A-123
a. Benthos A-123
b. Periphyton or Aufwuchs A-123
c. Plankton A-124
d. Nekton A-125
e. Bioassays and Biomom'toring A-125
f. Diversity Indices A-127
5. The Significance and Interpretation of
Biological Data A-128
6. Bibliography A-140
D. Engineering and Physics A-142
1. Mapping A-142
11
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Appendix
2. Flow A-144
a. Velocity A-145
b. Volume A-146
c. References A-146
E. Computer Applications A-148
1. Stream A-148
2. DIV A-154
3. DIVERS A-158
4. DPL0T A-161
5. ANALYZE A-165
6. STR-CLAS A-173
2. IMPLEMENTATION A-186
A. Cost A-186
B. Scheduling A-187
C. Motivation A-187
3. LIMITATIONS A-189
A. Time and Transportation A-189
B. Methods and Equipment A-189
C. Dealing with Owners A-190
4. EVALUATION A-191
5. BIBLIOGRAPHY A-193
A. Core References A-194
B. Additional References A-197
iii
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Appendix
C. Periodicals A-198
D. Movies A-199
E. Equipment A-204
6. WATER POLLUTION AND ENVIRONMENTAL GLOSSARY A-206
7. LABORATORY AND/OR FIELD SAFETY A-249
A. General Comments „ A-249
B. Bacterial Studies „ A-250
C. Chemistry „ A-251
D. Field Trips A-251
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Appendix I Water Quality Parameters
This appendix is included as a technical reference aid to teachers
using this guide. It is organized in four parts: (A) Chemistry,
(B) Bacteriology, (C) Aquatic Biology, and (D) Engineering and Physics.
A. Chemistry
Chemical parameters are quite specific, can be quantitated
relatively quickly and precisely, and can be related to water
quality requirements. It is seldom feasible or worthwhile to apply
all analytical procedures to a given water sample. However, certain
analyses are performed more or less routinely on water samples and
are included in this section.
Each part includes an identification of the selected parameter and
its common sources. This is followed by a description of the
chemistry involved in the more common approaches to the analysis of
that parameter. The procedures include references to commercial
testing kits and, in some instances, detailed instructions for
those who do not have access to commercial kits.
Commercial kits provide effective approaches to rapid and reasonably
accurate analyses, especially when time, facilities and lack of
trained personnel are limiting factors. Consequently, the procedures
include references to the following commercial units:
Delta Model 50 Portable Laboratory, Delta Scientific Corp.,
Lindenhurst, New York 11757
Hach DR-EL Portable Engineer's Laboratory, Hach Chemical Co.,
Ames, Iowa 50010
LaMotte Model #AM-21, LaMotte Chemical Products Co.,
Chestertown, Maryland 21620.
These kits have been identified only because they proved satis-
factory during the development of this program. This endorsement
does not imply superiority to other units that may be commercially
available.
An annotated bibliography appears at the end of this section. The
listings include those references which should be readily available
when investigating chemical parameters of pollution.
1. Acid-Base Parameters
a. Acidity
Acidity, a measure of the ability of a water sample to
neutralize hydroxide (OH~) ions, is subdivided into free
(mineral), un-ionized (weak acid) and total forms. The
chemical species which neutralizes the hydroxide ion is
A-l
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Appendix 1
identified as the hydrogen (H+) ion and is present in all
water samples.
Some of the substances which contribute to acidity (i.e.,
serve as sources of hydrogen ions) are depicted in Fig. 1.
Direct hydrogen ion donors are depicted within the circle
while those outside the circle provide hydrogen ions directly.
Fe*1* 3H40
Fig. 1 - Total Acidity
1) Free Acidity
All acids contain hydrogen; however, certain acid com-
pounds readily dissociate to form H ions in water
solution.^ This dissociated (free) form of the hydrogen
ion is known as free acid (Fig. 2) and is a component of
industrial wastes and drainage from sulfide-rich terrain.
Fig. 2 - Free Acidity vs. Total Acidity
1 +
The H is bound with water in forms such as H 0 but will be
considered as H+ throughout this guide. 3
A-2
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Appendix 1
When hydroxide ions are added to an acidic water sample,
they react with the free acid to form water, thus
resulting in a decrease in the free acidity (Fig. 3).
The quantity of hydroxide ions needed to reach the methyl
orange end-point is considered a measure of free acidity.
Fig. 3 - Free Acid Titration
a) Procedure
(1) The Hach and LaMotte kits do not provide
instructions for free acidity determinations.
However, it is possible to extend their C0?
procedures to include free acidity measure-
ments by titrating to a methyl orange end-
point before going on the phenolphthalein
end-point as follows:
1. Prepare the sample as described in Step 1
of the kit orocedures.
2. Add 1 drop of methyl orange indicator (see
(3) below). If the solution is orange-
yellow, the free acidity is not
measurable. Continue with step 2 of the
kit procedure if a bound acidity value is
desired.
A-3
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Appendix 1
3. Titrate with the titration reagent
designated for the kit's C02 procedure
until the orange-yellow end-point is
obtained. If chlorine residuals interfere
with the end-point determination, add 1
drop 0.1M sodium thiosulfate to a new
sample and repeat stages 2 and 3.
4. Record the volume of titrant used and
calculate the free acidity in the same
manner as described for the COo procedure.
Both kits use sodium hydroxide as the
titrant according to the following
reaction:
Na+ + OH" + H+ + X" = Na+ + X" + HgO
(X~ = any anion of a titrateable acid)
(2) The Delta kit has a free acidity procedure
which utilizes the reaction just described but
substitutes bromcresol green indicator for
methyl orange. It is also possible to adapt
the Delta C02 procedure to a free acidity
determination as described in (1).
(3) An alternate procedure to (1) is available as
follows:
Equipment:
25 ml graduated cylinder
medicine droppers
50 ml Erlenmeyer flask
burette or 1-ml pipette graduated in 0.1 ml units
Reagents:
Methyl Orange Indicator: Dissolve 0.5 g
methyl orange in 1 liter of distilled water.
0.1M Sodium Thiosulfate: Dissolve 2.5 g
Na2$20 -5 HpO in 100 ml of distilled water.
Q.02M NaOH: Prepare 1M NaOH by dissolving
4 g NaOH in 100 ml of FRESHLY BOILED distilled
water. Then dilute 2 ml of the stock solution
to a 100 ml volume with FRESHLY BOILED dis-
tilled water.
A-4
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Appendix 1
Method:
1. Measure 10 ml of sample into the 50 ml
Erlenmeyer flask.
2. Add 1 drop of methyl orange indicator.
If the solution is orange-yellow, the free
acidity is not measurable and should be
reported accordingly.
3. Titrate the sample with .02M NaOH. Record
the ml needed to reach the orange end-point.
If chlorine residuals interfere with the
end-point determination, add 1 drop 0.1M
sodium thiosulfate to a new sample and
repeat stages 2 and 3. (A reference for
the end-point can be prepared by adding 1
drop of methyl orange to 10 ml of pH 4.5
solution prepared by combining 1.36 g
0'3H0, sodium acetate, and 10 ml
1M HC2H302 with distilled water to make
100 ml solution.)
Calculations:
For uniformity, acidity is expressed as CaC03
equivalents, even though no CaCOj may be
present. The equation for calculating free
acidity is
mg CaC03/l = (A) (Molarity of NaOH) • 50,000
sample volume
If: Molarity of NaOH = 0.02M
Sample Volume = 10 ml
A = ml of 0.02M NaOH needed to attain the
methyl orange end-point,
Then: mg CaCO~/l = A x 100
O
2) Un-ionized (Bound) Acidity (CO Determination)
The acids in the larger circle of Fig. 2 account for un-
ionized acidity. However, C0? is primary contributor
to un-ionized acid levels in most samples
(H?0 + CO = H CO,). Carbon dioxide commonly enters
^ C. L, 3
A-5
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Appendix 1
water via absorption from the atmosphere and as an
end-product of both aerobic and anaerobid biological
oxidation and respiration.
i
Once the free acidity is decreased sufficiently by
reaction with hydroxide ions, weak acids such as carbonic
acid begin to release their hydrogen as free hydrogen ions
(Fig. 4).
Total Acidity
Fig. 4 - Weak Acid Titration
When enough hydroxide ions are added to reach the
phenolphthalein end-point, these substances will yield
most of their bound hydrogen. Consequently, a quantita-
tive evaluation of un-ionized acidity is achieved by
calculating the amount of hydroxide added.
a) Procedure
(1) The Hach, LaMotte, and Delta procedures are
actually evaluations of total acidity (free
and un-ionized). The free acidity in water
which has a pH greater than 4.5 is not
measurable. However, if the pH is less than
4.5, free acidity determinations must be com-
pleted and then subtracted from the total
value obtained by means of this procedure.
The reactions are:
2 NaOH + H0CO, = 2H.O + Na~CCL (1)
L. 3 ^ £ O
H?0 + NaJXJ + CO. = 2 NaHCO, (2)
c- f. 3 t- J
A-6
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Appendix 1
Reactions (1) and (2) are employed if NaOH
is the titrant. Only reaction (2) is employed
if Na2CO is the titrant.
«J
(2) The following procedure is suggested as an
alternate.
Equipment:
See (3) under Free Acidity.
Reagents:
Phenolphthalein Indicator: Place 0.5
g phenolphthalein in 50 ml denatured
ethanol and dilute to 100 ml with distilled
water.
0.1M Sodium Thiosulfate: Refer to Free Acidity,
part (3) for preparation.
0.02M NaOH: Refer to Free Acidity, part (3).
Method:
1. Measure 10 ml of the sample into the
50 ml Erlenmeyer flask.
2. Add one drop of phenolphthalein indicator.
(If the solution turns pink, there is no
measurable acidity.) Titrate with 0.02M
NaOH until the pink phenolphthalein end-
point is reached.
Calculations:
Acidity is expressed as mg/1 CaCO.,. The
un-ionized fraction may be calculated
according to the following equation:
mg CaC03/l = (B) (Molarity of NaOH) . 50,000
sample volume
If: Molarity of NaOH = 0.02M
Sample Volume = 10 ml
B = ml of 0.02M NaOH needed to attain
the phenolphthalein end-point after com-
pleting the methyl orange titration
A-7
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Appendix 1
Then: mg CaCCL/1 = B x 100
O
3) Total Acidity
Total acidity includes all hydrogen ion donors measured
by titration of a water sample to the phenolphthalein
end-point.
a) Procedure
(1) The CO- procedures in the Delta, Hach and
LaMotte kits will give total acidity
evaluations without modification.
(2) An alternate approach is to combine the
alternate procedures suggested for free un-
ionized acidity as follows:
1. Titrate to the methyl orange end-point
and calculate free acidity if desired
(see (1) on page A-6)
2. Add phenolphthalein and titrate to the
phenolphthalein end-point (see (2) on
page A-7)
Calculate the total acidity as mg
CaC03/l = C x 100, where C = total
volume of titrant used.
4) Reference
Standard Methods for the Examination of Water and Waste-
water, (12th ed.) American Public Health Association,
New York, 1965, pp. 46-47.
A-8
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Appendix 1
b. Alkalinity
Alkalinity is an indicator of the ability of a given water
sample to neutralize or accept hydrogen (H+) ions. Some
of the substances which comprise or contribute to alkalinity
within the pH range of 4.5 to 11 are depicted in Fig. 1.
The circle on the left of Fig. 1 includes several substances
which accept hydrogen ions directly during alkalinity
measurements (titrateable alkalinity). The circle on the
right includes substance which undergo chemical changes such
as the hydrolysis of water which produce hydrogen ion
acceptors. Those chemical species within the overlap of the
two circles may serve in both capacities. Hydroxide, car-
bonate, and bicarbonate ions are normally the predominating
members of their respective groups.
Fig. 1 - Components of Alkalinity
(pH 4.5 to 11)
Alkalinity is determined by titrating samples which are
alkaline to phenolphthalein to the phenolphthalein end-point
with sulfuric acid. This serves as a measure of the
"phenolphthalein alkalinity" which includes nearly all
hydroxides and half of the carbonates present. Titration is
then continued beyond the phenolphthalein end-point to the
methyl orange or bromcresol green-methyl orange. This step
of the titration neutralizes the remaining half of the
carbonates and the bicarbonates. The addition of the
sulfuric acid volume needed to reach the phenolphthalein
end-point to the amount needed to reach the methyl orange
end-point leads to a calculation of the "total alkalinity."
A-9
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Appendix 1
Sometimes it is desirable to attempt a calculation of the
concentrations of individual contributors to alkalinity.
Simplified calculation procedures summarized in Table 1
are based upon the following concepts:
(1) Hydroxides, carbonates, and bicarbonates are
usually the major sources of alkalinity in
natural waters.
(2) Hydroxides and bicarbonates are incapable of
existing together in the same solution.
(Assumed, but not true.)
(3) The hydroxide supply is essentially exhausted by
titration to the phenolphthalein end-point.
(4) One-half of the carbonates is titrated upon
reaching the phenolphthalein end-point.
(5) The bicarbonates and the remaining half of the
carbonates are titrated when proceeding from the
phenolphthalein end-point to the methyl orange
end point.
Table 1. Alkalinity Relationships
TITRATION
RESULT
P=T
P< 1/2T
P=1/2T
P> 1/2T
P=0
HYDROXIDE
ALKALINITY
equals T
0
0
T -2 (T-P)
or 2P-T
0
CARBONATE
ALKALINITY
0
2P
T
2(T-P)
0
BICARBONATE
ALKALINITY
0
T-2P
0
0
T
P = Phenolphthalein Alkalinity T = Total Alkalinity
1) Procedure
a) Refer to Delta, Hach, and LaMotte kits. The
reactions are classical acid-base neutralizations.
H+ + X" = HX (X" = any anion of a weak acid)
A-10
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Appendix 1
b) The following procedure using available
laboratory materials is suggested (1).
Equipment:
25 ml graduated cylinder
medicine droppers
50 ml Erlenmeyer flask
burette or 1 ml pipette graduated in 0.1 ml units
Reagents:
Methyl Orange Indicator: Dissolve 0.5 g of
methyl orange in 1 liter of distilled water.
Phenolphthalein Indicator: Place 0.5 g of
phenolphthalein into 50 ml of denatured ethanol
and dilute to 100 ml.
0.1M Sodium Thiosulfate: Dissolve 2.5 g of
Na?S 0,'5H?0 in 100 ml of distilled water.
£ C. *J ^~
0.01M Sulfuric Acid: Add 3 ml of concentrated
H2SO (18M) to 1 liter of distilled water,
yielding 0.05M H2S04. Dilute 20 ml 0.05M H2S04 to
100 ml yielding 0.01M H2S04.
Method:
1. If present, remove free residual chlorine
by adding 1 drop of sodium thiosulfate to
a 100 ml sample.
2. Measure a 10 ml sample into the titration
flask and add 1 drop of phenolphthalein. If
solution is not pink, no free alkalinity is
present. Skip step 3 and proceed to step 4.
3. Add 0.01M sulfuric acid to the sample with
the pipette or burette. Record the number of mis
needed to reach the pink end-point. Use this
number in the calculation of phenolphthalein
alkalinity.
4. Add 1 drop of methyl orange indicator.
A-11
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Appendix 1
5. Continue to titrate with 0.01M sulfuric acid
until the methyl orange end-point is reached.
Record the volume (ml) used and combine this
value with the volume (ml) obtained in step 3.
Use this value for the calculation of total
alkalinity. (A reference for the end-point
can be prepared by adding 1 drop of methyl
orange to 10 ml of pH 4.5 solution prepared by
combining 1.36 g NaC^Cyi^O and 10 ml 1M
HCpHoO with water to make 100 ml solution.)
Calculations:
For uniformity, alkalinity is expressed as mg
CaCOo/1 even though there may be no CaCOo present.
The equation for the phenolphthalein alkalinity is
mg CaC03/l = A x (Molarity of H2SO ) x 100,000
Volume of Sample
where A equals ml of the titrant used to reach the
phenolphthalein end-point and the concentration
of the sulfuric acid is expressed as molarity.
This can be reduced to:
mg CaC03/l = A x 100
if a 10 ml sample is used and the sulfuric acid
is 0.01M.
In the same way, the total alkalinity is calculated
as
mg CaC03/l = B x
100
where B is the TOTAL number of ml needed to reach
the methyl orange end-point.
2) Reference
Standard Methods for the Examination of Mater and Waste-
water, (12th ed.), American Public Health Association,
New York, 1965, pp. 48-50.
A-12
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Appendix 1
c. pH
pH is a measurement which reflects the instantaneous
free hydrogen ion concentration in a water sample. Free
hydrogen and hydroxide ions exist in equilibrium in all
aqueous solutions. If these ions are present in equal
amounts, the sample is described as neutral and has a pH
value of 7. If the hydrogen ion concentration is less than
the hydroxide ion concentration, the solution is said to be
basic and has a pH value greater than 7. If the hydrogen
ion concentration is greater than the hydroxide ion con-
centration, the solution is acidic and has a pH value less
than 7 (Fig. 1).
acidic basic
[H+]> [OH"] [H+] < [OH"]
10 11 12 13 14
neutrality ( [ H+ ] = [ OH" ] )
Fig. 1 - pH Relationships
It is essential to regognize that pH is not a measurement
sensitive to the presence of substances which may con-
tribute to the total acidity or alkalinity of a given
sample. Consequently, it must not be confused with the
results of total acidity and alkalinity determinations.
Samples which possess a neutral pH may possess high acidity
and/or alkalinity values. Because natural waters are
buffered by the C02, HC03, CO system to a pH range of
6.5 to 7.5, marked deviations from neutrality are generally
the result of industrial or acid mine contamination.
The pH of water samples is usually determined by either
colorimetric or electrometric techniques. Colorimetric
procedures rely upon chemical substances which undergo
color changes with change in pH. There are numerous
reagents which demonstrate this phenomenon; however, each
is effective as a pH indicator within a limited pH range
only. A versatile pH measurement system must contain
numerous indicators covering the entire pH spectrum.
These indicators are either impregnated on paper strips,
A-13
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Appendix 1
used separately in solution form, or combined to create
a "universal" or "wide-range" indicator solution.
Electrometric techniques yield the greatest accuracy. They
employ meters which, by means of a glass electrode, detect
differences in electric potential which occur with differing
pH values. Once the meter is properly calibrated, pH read-
ings are read directly from the instrument scale.
Procedure:
1) Refer to the Delta, Hach or LaMotte kits. All three
utilize colorimetric procedures.
2) As alternatives to the kits, the following procedures are
recommended:
a) Purchase universal indicator or a good quality pH
paper from any chemical supply house and use accord-
ing to the accompanying instructions.
b) Use a pH meter. Models differ in operation; there-
fore, instructions for their use must be obtained from
the manufacturer. The pH of a given sample should be
obtained propmptly to prevent changes due to reactions
with COg from the air or loss of 062 to the air.
2. Dissolved Gases
a. C02 - See end of this reference section for material.
b. Chlorine (residual)
Both free and combined forms of chlorine are used as disin-
fectants in attempts to curb waterborne diseases. Chlorine
does not occur naturally in water but may enter through
sewage treatment effluents and industrial wastes.
In the quantitative determination of chlorine, an organic
compound orthotolidine is oxidized in acid solution by both
free and combined forms of chlorine. This produces a yellow
colored compound hoioquinone, which is measured colorimetrically.
An alternate method which corrects for color interferences
is known as the orthotolidine-arsenite method. Total resi-
dual chlorine is measured in the usual way with orthotolidine
as described above. A second test which serves as a blank
is prepared by introducing sodium arsenite solution before
adding the orthotolidine. The arsenite, being a much stronger
reducing agent than orthotolidine, reduces both free and
A-14
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Appendix 1
combined chlorine. This prevents their reaction with the
orthotolidine. Any color present in this second test is
due to interference by other chemical substances and the
reagents being used. The total residual chlorine level can
be calculated as follows:
Total chlorine
residual
(Test 1)
1) Procedure
and
Interfering
Color
(Test 2)
Total Residual
Chlorine
a) Refer to Delta kit. For clear waters, Delta uses
the orthotolidine method. The reaction is as
follows:
CH3
CH,
H
orthotolidine
A holoquinone (yellow)
For turbid waters, Delta uses the orthotolidine-
arsenite method as described above.
b)
c)
Refer to Hach kit. Because of color fading,
has developed a modified orthotolidine reagent called
0-ToliVer which stabilizes the final color for longer
periods of time. The reaction is similar to that in
the Delta kit.
Refer to the LaMotte kit. It uses the orthotolidine
method as described above.
c. Dissolved Oxygen
Dissolved oxygen is an essential substance for the support of
most aquatic life. Its concentration in water (normally very
low compared with that in air) varies with fluctuations in
such factors as temperature, types and concentrations of dis-
solved and suspended solids, biotic activity, and agitation
of the water. Both depressed and elevated (supersaturated)
dissolved oxygen levels are encountered in aquatic studies.
In view of our understanding of the biological role of DO,
deleterious effects of low or nonexistent levels of DO are
hardly surprising. Harmful effects accompanying DO super-
saturation of water supplies have not been so readily
A-15
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Appendix 1
anticipated. However, fish have demonstrated low tolerance
to DO supersaturation as indicated by an increased incidence
of mortality and disease in such waters (1,2).
Regardless of the test used for determination of DO, the
sampling procedures must avoid aeration and warming. More-
over, the test must be done immediately or the oxygen must
be fixed if chemical and biochemical influences are to be
avoided. The Azide-Winkler method, an accurate and feasible
test for DO, eliminates interference by nitrite ions through
the use of sodium azide. Dissolved oxygen is fixed by the
addition of manganese sulfate and an alkali-iodide-a/:ide
reagent. In this reaction, the oxygen oxidizes manganous
ions to manganese oxyhydroxide; Mn 0(OH)2. Under acid con-
ditions (obtained by adding concentrated suKuric acid or
the less dangerous solid form of sulfamic acid), the man-
ganese oxyhydroxide oxidizes iodide ions to produce free
iodine. The amount of free iodine produced is equivalent
to the dissolved oxygen originally present. Following
titration to a pale straw color with sodium thiosulfate, starch
is added and the titration is continued until the blue color
disappears. With clean water samples, the titration may be
delayed under acid conditions for up to 6 hours. Prompt
titration is required for polluted water.
1) Procedure
a) Azide-Winkler method (in lab without kit)^
(1) Equipment
4-5 ml pipettes
burette, in 0.1 ml units with a 50 ml capacity
BOD bottles, 300 ml capacity
Erlenmeyer flask, 250 ml
(2) Reagents
Manganese Sulfate Solution: Dissolve 480 g
MnS04-4H20 in distilled water, filter and dilute
to 1 liter.
Alkali-iodide-azide Reagent: Dissolve 500 g
NaOH and 150 g KI in distilled water and dilute
to 1 liter. To this solution add 10 g NaNj
dissolved in 40 ml of distilled water. This
reagent should not give a color with starch
solution when diluted and acidified.
A-16
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Appendix 1
Concentrated Sulfuric Acid: Use concentrated
reagent grade acid ((^$04). Handle carefully!
Starch Solution: Prepare a paste of 5-6 g soluble
starch in a small amount of distilled water.
Pour this paste into 1 liter of boiling distilled
water, allow to boil a few minutes and let settle
over night. Use the clear supernate.
Sodium Thiosulfate Solution: Dissolve 24.82 g
Na2s2°3'5H2.9 i" boi1ecl and cooled distilled
water and dilute to 1 liter. Preserve by add-
ing 0.4 g of NaOH per liter.
Working Sodium Thiosulfate Titrant 0.0375M:
Prepare by either diluting 375 ml sodium thiosul-
fate stock solution to 1 liter or by dissolving
9.30 g Na2S203-5H20 in freshly boiled and cooled
distilled water and dilute to 1 liter. (For
standardizing the sodium thiosulfate, refer to
Standard Methods, p. 407.)
b) In the field
(1) Fill a 300 ml glass stoppered bottle with sample
water by allowing the sample to enter through a
glass or rubber tube which extends to the bottom
of the bottle. An overflow displacing the bottle
contents 2-3 times is necessary to ensure that the
test sample has not been exposed to the air.
Stopper the bottle immediately upon removing the
tube. Be sure that no bubbles are trapped within
the bottle.
(2) Add 2 ml manganese sulfate to the collecting
bottle by means of a pipette inserted just below
the surface of the liquid.
(3) Add 2 ml alkali-iodide-azide reagent in the same
manner.
(4) Stopper with care to exclude air bubbles and mix
by inverting the bottle several times. When the
precipitate settles shake again and allow to settle.
Note: The oxygen is fixed according to the
following reaction.
Mn++ + 2 OH" + 1/2 02 —> MnO(OH)2 (eq. 1)
(golden brown
flocculant)
A-17
-------
Appendix 1
c) In the lab
(1) Add 2.0 ml concentrated H2S04 with the pipette
above the surface of the liquid; stopper and
invert several times to dissolve the precipitate.
Note: With the addition of sulfuric acid,
the proper low pH conditions are obtained
for the destruction of interfering NO'?" by
the sodium azide which was added in th~e alkali
iodide-azide reagent above. The following
reactions occur: ,
NaN3 + H+ —> HN3 + Na+ (eq. 2)
HN3 + N02- + H+—> N2 + N02 + H20 (eq. 3)
Under the same pH conditions, the Mn+'*
oxidizes I" to produce free I2 as follows:
Mn 0(OH)? + 2 I" + 4 H+ —> Mn
3H20
++
+ I,? +
teq. 4)
(2) In an Erlenmeyer flask, titrate the 300 m'l sample
with 0.0375M sodium thiosulfate to a pale straw
color.
(3) Add 2 ml of starch solution. A blue color forms
indicating the presence of molecular iodine, I^.
Continue titrating until the molecular iodine Ts
reduced to iodide ions as indicated by the dis-
appearance of the blue color.
Note: The reaction is
2 S203" + I2—> S406" + 2 r (eq. 5)
(4) Record the total amount of sodium thiosulfate used.
2) Calculations
1 ml of 0.0375M Na2So03 is equivalent to 0.2 mg DO per
300 ml sample as follows:
According to (eq. 5), S203~~ loses 1 electron so that 1
liter of 0.0375M Na2$203 will lose 0.0375 moles of electrons
(or 1 ml will lose 3.75 x 10~5 moles of electrons). To
change 1 mole of molecular oxygen (02) to 0" requires
4 moles of electrons. 3.75 x 10"5 moles of electrons will
reduce approximately 9.4 x 10~& moles of molecular 02.
A-18
-------
Appendix 1
Since 1 mole of CL has a mass of 32 g, 9.4 x 10 moles
has a mass of O.OoOS g or 0.3 mg. Each milliliter of
sodium thiosulfate used in the titration of a 300 ml sample
indicates the presence of 0.3 mg 02/300 ml or 1 mg
liter (1 ppm).
Summarizing: Each ml of sodium thiosulfate added in steps
3 and 4 equals 1 mg/1 DO (1 ppm).
3) As an alternative to the laboratory method described above,
refer to either the Hach or LaMotte kits. They utilize
chemical principles outlined for the laboratory method
with exceptions as follows:
a) Hach: Substitutes phenylarsene oxide (PAO) for the
sodium thiosulfate titrant.
b) LaMotte: Utilizes an unmodified Winkler procedure;
consequently, it is subject to interference by nitrite
ions.
4) References
(1) McKee, J. E. and H. W. Wolf, Water Quality Criteria.
(2nd ed.), State Uater Quality Control Board,
Pub. #3-A, Sacramento, Calif., 1963, p. 181.
(2) Ibid.
(3) Standard Methods for the Examination of Water and
Wastewater, (12th ed.), American Public Health
Association, New York City, 1965, pp. 415-419.
(4) Carbon Dioxide prior to Chlorine
Refer to Section 2), Un-ionized (Bound) Acidity,
Acid-Base Parameters (see page A-5).
3. Dissolved and Suspended Solids
a. Chloride
The chloride ion is a component of many salts and most living
organisms. Because chloride salts are usually soluble, ions
find their way into natural waters by phenomena such as ero-
sion and leaching. Examples of other common chloride sources
include sea water intrusion, human and animal sewage, fertil-
izers, industrial wastes, and winter salting of highways.
Gradually add mercuric nitrate or silver nitrate solution to a
water sample containing an indicator. The mercuric or silver
ions combine with the chloride ions until the chloride supply
A-19
-------
Appendix 1
is essentially depleted. At this point, mercuric or silver ions
form a colored complex by reacting with the indicator. The
amount of mercuric nitrate or silver nitrate solution added
indicates the chloride ion concentration.
1) Procedure
a) Refer to the Hach, Delta, or LaMotte kits if they are
available. These kits utilize the following reactions:
Hach Kit: (1) Hg++ + 2CT = HgCl2
(2) Hg++ + Diphenylcarbazone = purple complex
Delta & LaMotte Kits:
(1) Ag+ + CT = AgCl
(2) 2 Ag+ + Cr04 = Ag2Cr04 (red color)
b) If a commercial kit is not available, the following
procedure which uses the above reactions is suggested.
Equipment:
burette, 25 ml
porcelain evaporating dish, 250 ml
glass stirring rod
assorted beakers, graduates, one-liter volumetric
flasks, and bottles as needed
five ml pipette
Reagents:
Silver Nitrate, 0.0141M: Dissolve 2.396 g silver nitrate
(AgNOo) in distilled water and dilute to 1 liter in a
volumetric flask. Standardize against 0.0141M sodium
chloride solution. One ml silver nitrate solution
equals approximately 0.500 ml Cl".
Sodium Chloride, 0.0141M: Dissolve 0.8241 g of sodium
chloride (NaCl) in distilled water and dilute to 1 liter
in a volumetric flask. One ml sodium chloride solution
equals 0.500 mg Cl~.
A-20
-------
Appendix 1
Potassium Chromate Indicator: Dissolve 50 g potassium
chromate (I^CrC^) and dilute to 1 liter with distilled
water.
Method:
1. To standardize the silver nitrate, add 20 ml of
0.0141M sodium chloride to 1 ml of potassium
chromate indicator in a porcelain evaporating dish.
Titrate as per step 4 below. Then calculate the
normality constant as follows:
m1 AgN°3 x 500 = Normality constant
20
2. Place 100 ml sample or a smaller quantity diluted
to 100 ml with distilled water in a porcelain
evaporating dish.
3. Add one ml of potassium chromate indicator with a
pipette.
4. Add silver nitrate solution from a burette, stirring
the dish contents until a uniform pinkish-yellow
end-point is reached. Record the ml of silver
nitrate added.
5. Repeat steps 2, 3, and 4 above using 100 ml of
distilled water as a blank in place of the sample.
6. Calculate the final result as follows:
mg/1 Cl~ = (ml AgN03 for sample - ml AgN03 for blank) (Normality Constant)
ml original sample
2) Reference
Water Pollution Control Federation, Simplified
Laboratory Procedures for Wastewater Examination,
WPCF Publication, No. 18, 1968, pp. 45-46.
b. Hardness—Calcium, Magnesium, Total
Hardness is a water quality parameter which limits the lathering
or foaming ability of soaps and increases the tendency of a
water sample to produce scale in pipes, heaters, and boilers.
Hard water is caused by the presence of divalent ions such as
A-21
-------
Appendix 1
Calcium (Ca ) and magnesium (Mg ). Additional ions (e.g.,
Sr , Mn , Fe++) can cause hardness but are present only in
limited amounts in most water supplies. If their concentra-
tions are elevated, they should be included in calculations of
total hardness. All of these cations enter water sources
via industrial wastes, sewage, and contact with soil and rock
formations.
The chemical determination of total hardness involves the
titration of a water sample to which an indicator, such as
Eriochrome black T, has been added. The substance EDTA is
used as the titrant because of its ability to complex with
divalent cations. Prior to titration, the indicator forms a
red complex with Ca++ or Mg++. During titration within a
specific pH range, the red indicator releases its bound
cations to the EDTA and reverts to its blue pigment. Total
hardness is calculated from the amount of EDTA needed to
reach the blue end-point.
In the determination of calcium hardness, magnesium is pre-
cipitated as magnesium hydroxide by the addition of alkali.
The rest of the procedure is completed as outlined above.
Magnesium hardness is calculated by subtracting the calcium
value from the total hardness figure.
1) Procedure
a) For total hardness, refer to the Hach or LaMotte
kits. The following reactions are employed:
M++
(Eriochrome black T) \ ^ (M Eriochrome black T)
blue color I7 ~/^~~ wine red complex
EDTA _A > (M-EDTA)
colorless complex
M++ = any divalent cation
EDTA = ethylenediamine tetraacetic acid
b) For calcium hardness, refer to tha Hach, LaMotte or
Delta kits. Following the addition of sodium hydroxide
or potassium hydroxide to precipitate magnesium
hydroxide, the following reaction occurs:
Mg++ + 2 OH" = Mg (OH)2
The reactions obtained from the Hach and LaMotte kits
were described above. While the Delta kit uses different
A-Z2
-------
Appendix 1
reagents, it appears to utilize a similar process.
c) Magnesium hardness may be calculated by determining
the difference between the total hardness and calcium
hardness value.
d) As an alternate procedure, calcium hardness may be
evaluated in a rough quantitative fashion by the follow-
ing precipitation procedure according to the reaction:
Ca++ + CyOfl— * CaC?04 (s)
£ " t*T/ T • j \
(solid)
Equipment:
2 test tubes
4 dropping pipettes
Reagents:
Stock 0.01M Ca++ Solution: Add 1.11 g of CaClo to
100 ml of distilled water and dilute to 1 liter.
Working Ca++ Standard (80 ppm Ca++): Add 20 ml of
the stock solution to 80 ml of distilled water.
Concentrated Ammonia Water.
4% Ammonium Oxalate: Dissolve 4 g of (NHJoCpO/ in
50 ml of distilled water and dilute to 100 ml.
Method:
1. Prepare a reference sample containing Ca++ by placing
20 drops (1 ml) of the working Ca standard into
Tube 1.
2. Place 20 drops of the water sample into Tube 2.
3. Add two drops of concentrated ammonia water to
both tubes.
4. Add 4% ammonium oxalate dropwise until a reaction
is observed. Do not add more than 5 drops.
5. Compare the amount of precipitation in Tube 2 with
that in Tube 1. Report your result as being
greater than or less than 80 ppm.
A-23
-------
Appendix 1
2) Reference
Standard Methods for the Examination of Hater and
Wastewater, (12th ed.). American Public Health
Association, New York City, 1965, p. 149.
c. Iron
Ionic forms of iron occur in water as either the iron (II) or
iron (III) form. Iron (II) is easily oxidized to iron (III)
which reacts with hydroxides to form insoluble iron (III)
hydroxide, thus keeping iron concentrations in most water
supplies at low levels. While toxic to many organisms,
elevated iron concentrations support iron bacteria (which may
cause corrosion) in pipe lines or structures with formation
of slimes, pits, encrustations, and other undesirable effects.
Dissolved iron originates from soils or rock formations during
leaching and erosion processes effected by acidic water flows.
Also, there is evidence which suggests that iron enters water
sources through changes produced in environmental conditions
as a result of biological reactions.
In quantitative iron studies, it is necessary to convert all
of the iron (III) to the soluble iron (II) form. This; is
accomplished by dissolving any precipitated iron (III) hydroxide
by the addition of hydrochloric acid and reducing the iron (III)
species to iron (II) through the action of hydroxylamine, a
strong reducing agent. The water sample is then treated with
1 ,10-phenanthroline which combines with the iron (II) to form
an orange-red complex suitable for colorimetric evaluation.
An alternative procedure involves the conversion to iron (II),
as described, followed by the addition of ethyl enedi ami ne
which buffers the water sample and complexes* heavy metals
which might give erroneously high results. 2,2,2-tripyridine
is added to yield a reddish-purple iron (II) complex for color-
imetric study.
1) Procedure
Refer to the Hach, Delta, or LaMotte kits. The following
reaction sequences are used:
Fe(OH)3
4 Fe3+ + 2 NH2OH = 4 Fe++ + N20 + H20 + 4 H+
* binds up
A-24
-------
Appendix 1
These are followed by:
1. In the Hach and LaMotte kits
+ Fe1
2. In the LaMotte kit:
_ ft
+F.
«
2) Reference
Sawyer, C. N.,and P. L. McCarty, Chemistry for
Sanitary Engineers, (2nd ed.), McGraw-Hill Book
Co., New York City, 1968, pp. 446-448.
d. Nitrate
Nitrate ions are end-products of the oxidation of nitrogen
or nitrogen compounds. They are formed by (1) the nitrogen
fixation activity of certain bacteria and algae, (2) the
oxidation of atmospheric nitrogen during electrical storms, and
(3) the oxidation of nitrogenous compounds (ammonia, nitrates,
proteins, certain organics) in both water sources and aerobic
sewage treatment systems. Their use in fertilizers as a source
of nitrogen for plant protein synthesis constitutes a source
of pollution, as excess amounts are carried into water supplies
by percolation and runoff.
In the suggested procedures, nitrates are measured by reduction
by cadmium to nitrite ions followed by reaction with sulfanilic
acid to form a diazonium salt. The salt is reacted with 1-
naphylamine hydrochloride to form a red-colored azo dye.
The presence of nitrite ions in the original water sample will
cause falsely high nitrate values. A correction is achieved
by measuring the nitrite level separately (see Nitrite) and
subtracting the resulting nitrite value from the nitrate value
obtained in the cadmium reduction method just described.
1) Procedure
Refer to the Hach, Delta, or LaMotte kits.
A-25
-------
Appendix 1
Their reactions are:
N03" + Cd = N02~ + CdO (reduction of N03")
+ HN02 + HC1 =
Nitrous acid
Sulfanilic
acid
(Eq. 1)
S03H
+ 2 H20 (Eq. 2)
(diazotization)
A diazonium
salt
Diazonium
salt
NH3C1
1-Naphthylamine
hydrochloride
(Eq. 3)
NH3C1
A red-colored
azo dye
e. Nitrite
Nitrites are intermediates in the chemical or biological
modification of nitrogenous compounds such as ammonia, nitrates,
certain organics, dyes, and proteins. Accordingly, they may
occur in water supplies containing such substances.
Nitrites are measured by conversion to a diazonium salt through
reaction with sulfanilic acid. Upon reaction with 1-riaphthyla-
mine hydrochloride, a red-colored dye develops which is easily
measured by colorimetric procedures.
Procedure
Refer to the Hach, Delta, or LaMotte kits. The chemistry is
described by Equations (2) and (3) of the Nitrate Ion section.
f. Phosphate
The phosphate ion exists in both organic and inorganic forms.
With the exception of bottom sediments, and samples containing
algae and suspended particles which may possess organic phos-
phorous as a major phosphorous form, emphasis is placed on
analystical evaluations of the inorganic forms outlined in
Table 1.
A-26
-------
Appendix 1
Table 1. - Inorganic Phosphates
Polyphosphates* Orthophosphates*
(meta) (MP03)x MH2P04
(pyro) M4P207 M2HP04
(tri) M5P3010 M3P04
(tetra) MgP4013
* M = any monovalent cation
These determinations are considered significant because of
our increased awareness of the role of phosphates in life
processes (ATP, enzyme function, buffering) combined with
their extensive use in fertilizers, detergents, water sof-
teners and as nutrients in the biological degradation of
sewage.
The suggested procedures detect only orthophosphates; con-
sequently, it is necessary to convert the polyphosphates to
the ortho form if a reliable measure of the inorganic
phosphates content is to be obtained. This process occurs
in all aqueous systems but may take from hours to several
days for completion under field conditions. In the labora-
tory, the conversion is hastened by boiling the sample in an
acidic solution. If organic phosphorous is to be included
in the analysis, it must be converted to the orthophosphate
form through oxidation by sodium persulfate (refer to Stand-
ard Methods).
Detection of the orthophosphate form is accomplished by
reacting it with ammonium molybdate to form ammonium phos-
phomolybdate. This product is subsequently reduced to
molybdenum blue by reaction with stannous ions.
1) Primary Procedure
Refer to the Hach,.Delta, or LaMotte kits if they are
available.
a) P04"3 + 12 (NH4)2 Mo04 + 24 H+ = (NH4)3P04'12Mo03
+ 21 NH4++ 12 H20
b) (NH4)3P04'12Mo03 + Sn++ = (molybdenum blue)
+ Sn+4
A-27
-------
Appendix 1
2) The following alternative procedure for orthophosphate
only is suggested. The reactions just described are utilized.
Equipment:
2 test tubes
3 medicine droppers
Reagents:
Stock O.OQ1M Phosphate Solution: Add 0.136 g KH2P04 to
distilled water making total volume 1 liter.
Working Standard ppm ^PO/p Add 10 ml of stock solution
to yyo mi of distilled water.
Ammonium Molybdate - Nitric Acid Reagent: Dissolve 15 g
of ammonium molybdate in 300 ml of distilled water. Add
100 ml of nitric acid 1:1 dilution of concentrated HNC>3
and saturate with ammonium nitrate.
Method:
1. Prepare a reference sample containing phosphate ions
by placing 20 drops of working standard in Tube 1.
2. Place 20 drops of the water sample into Tube 2.
3. Add 10 drops of the ammonium molybdate-nitric acid
reagent to each tube.
4. Add a few crystals of stannous chloride to both
tubes. A blue color should appear if orthophosphate
ions are present.
5. Compare the intensity of the blue pigment in Tube 2
with that of Tube 1. Report your results as having
less than or greater than 1 ppm orthophosphate.
Sulfate
The sulfate ion, a complex of sulfur and oxygen, is capable of
serving as an oxygen donor for biochemical oxidations occur-
ing under anerobic conditons. This action results in the
conversion of the sulfate ion to the sulfide form which
equilibrates with hydrogen ions to form hydrogen sulfide.
The latter substance possesses an objectionable "rotten egg"
odor and is capable of being oxidized by sulfur bacteria to
form sulfuric acid. The sulfate ion is derived from sewage,
A-28
-------
Appendix 1
industrial and agricultural effluents, erosion, and percola-
tion of water through pyrite or sphalerite ore deposits.
Analytical techniques for sulfates are based upon the forma-
tion of insoluble barium sulfate by the addition of barium
ions. The resulting solid may be collected, dried, and weighed
or may be kept in colloidal suspension by the use of a con-
ditioning reagent containing hydrochloric acid, sodium chloride,
glycerol, and other organic compounds and then measured by
turbidimetric procedures. At least one titrimetric procedure
is available which involves the gradual addition of barium
chloride to a water sample containing an indicator. The
barium ions precipitate with the sulfate ions until the
sulfate ion supply is essentially depleted. Excess barium
ions then combine with the indicator to produce a color
change. The sulfate level is calculated from the amount of
barium chloride needed to achieve the end-point.
1) Procedure:
a) Refer to the Hach or Delta kits. They utilize the
following reactions:
(1) Hach kit (Turbidimetric Procedure)
Ba++ + S04" = BaS04 (solid)
(2) Delta kit (Titrimetric Procedure)
Ba++ + S04" = BaS04 (solid)
THEN Ba++ + Indicator = orange-red complex
b) An alternative rough quantitative procedure is
suggested as follows.
Apparatus:
2 test tubes
4 medicine droppers
Reagents:
Stock 0.01M MnS04 Solution: Add 1.7 g MnS04'H20 to
100 ml distilled water and dilute to 1 liter.
Working S04~" Standard (96 ppm S04"): Add 10 ml of
stock solution to 90 ml of distilled water.
A-29
-------
Appendix 1
6M Hydrochloric Acid: Dilute concentrated HC1 (12M)
to 1/2 its original concentration.
0.1M Barium Chloride: Dissolve 2.08 g BaCl2 in 50 ml
of distilled water and dilute to 100 ml.
Method:
1. Prepare a reference sample containing SO^ by
placing 10 drops of the working standard into
Tube 1.
2. Place 10 drops of the water sample in Tube 2.
3. Add 2 drops of HC1 and 1 drop of BaCl2 to each
test tube.
4. Formation of a white precipitate or cloudiness
indicates the presence of SO*". Compare the
amount of cloudiness or precipitation in Tube 2
with that in Tube 1 and report your result as
greater or less than 96 ppm S0,~~-
h. Turbidity
Turbidity limits light penetration within a body of water by
causing incident light to be scattered or absorbed rather
than transmitted appreciable distances through the sample.
Turbid water is caused by the presence of suspended organic
and inorganic solids derived from erosion, surface drainage
systems, and domestic and industrial wastes. It exerts a
negative influence on photosynthesis and water temperature
by reducing the amount of light reaching subsurface areas and
can, by itself, kill fish and other organisms. Increases in
turbidity may follow a chain reaction sequence by providing
bacteria and other microorganisms contributing to turbidity
with an abundant supply of nutrients required for growth and
reproduction.
Turbidity is measured by comparing the interference to the
passage of incident light in the questioned sample with that
in a standard reference. Although the accuracy of photometric
or nephelometric techniques is questionable, such procedures
are convenient for approximating turbidity and are used in
most commercial kits.
Procedure:
1) Refer to Hach or Delta kits.
2) The following procedure which measures the depth of light
A-30
-------
Appendix 1
penetration can be used to supplement photometric deter-
mination of turbidity. The depth of light penetration
is affected by turbidity, but also color.
a) Equipment (a home-made Secchi disk costs about $.50)
(1) calibrated rope
(2) tempered plywood aide, 20 cm in diameter, with
alternate white and black quadrants
(3) eye-bolt, washers
20 cm
Calibrated rope
-Washer
Figure 1.
b) Method
(1) Lower disk and record depth of disappearance.
(2) Lower disk below the recorded point and then
slowly raise it. Record the depth at which the
disk first becomes visible.
(3) Average the two readings. Secchi disk readings
range from a few centimeters to over 40 meters.
4. Oxygen Demand
Biochemical Oxygen Demand (BOD)
BOD values reflect the quantity of molecular oxygen required
for the decomposition of organic compounds by aerobic biochem-
ical processes. Consequently, BOD values serve as an index of
the pollution strength of wastes by measuring the amount of
oxygen which may be removed from water supplies as these
wastes are being aerobically stabilized.
A-31
-------
Appendix 1
The BOD determination is a bioassay procedure requiring
(1) excess 0?, (2) favorable physical conditions, (3) essential
nutrients, (4) suitable organisms, and (5) time. While 20
days are usually required to approach complete waste stabiliza-
tion, the length of the assay is set at 5 days. The shorter
period usually allows for the measurement of a substantial
fraction of the total BOD. It also minimizes interference by
autotrophs, particularly nitrifying bacteria, which aerobically
metabolize inorganic nitrogen. These organisms usually require
more than 5 days to become established in a fresh sewage
sample but may start promptly in a stream, lake, or effluent
sample. Aerobic stabilization of inorganic nitrogen does
create an increased oxygen demand; however, attempts to
evaluate this parameter according to the following procedures
are not valid.
Aerobic stabilization of nitrogen components is becoming in-
creasingly important in impounded waters but is not normally
included in the described procedure.
1) Procedure for unchlorinated water
If the sample has been chlorinated, it is recommended
that the BOD not be performed. A good job of chlorina-
tion renders the BOD meaningless. However, a dechlorina-
tion and reseeding procedure is described in the next sec-
tion for those who desire to attempt it.
Equipment:
burette, graduated in 0.1 ml units with a 50 ml capacity
BOD bottles, 300 ml capacity
Erlenmeyer flask, 250 ml
10 ml measuring pipette
large-tipped volumetric pipette
incubator, controlled at 20° C
Reagents:
Manganous Sulfate Solution: Refer to the procedure for DO.
Alkaline Iodide-Sodium Azide Solution: Refer to the procedure
for DO.
Sulfuric Acid: Use concentrated reagent-grade acid (^SO.).
Handle carefully, since this material will burn hands and
A-32
-------
Appendix 1
clothes. Rinse affected parts with tap water to prevent
injury.
Sodium Thiosulfate Solution: Refer to the procedure for DO.
Starch Solution: Refer to the procedure for DO.
Distilled Water: Water used for solutions and for prepara-
tion of the solution water must be of highest quality. It
must contain no copper or decomposable organic matter.
Ordinary battery distilled water is not good enough.
Phosphate Buffer Solution: Dissolve 8.5 g KHgPO^., 21.75 g
K2HP04, 33.4 g Na2HP04-7H20 and 1.7 g NfyCl in distilled
water and make up to T liter. The pH buffer should be
checked with a pH meter (or pH paper).
Magnesium Sulfate Solution: Dissolve 22.5 g MgS04'7H20 in
distilled water and make up to 1 liter.
Calcium Chloride Solution: Dissolve 27.5 g anhydrous
CaCl2 in distilled water and make up to 1 liter.
Ferric Chloride Solution: Dissolve 0.25 g FeC^-GF^O in
distilled water and make up to 1 liter.
Dilution Water: Add 1 ml each of phosphate buffer, mag-
nesium sulfate, calcium chloride, and ferric chloride
solutions for each liter of distilled water. Store at
a temperature as close to 20 C as possible. This water
should not show a drop in DO of more than 0.2 mg/1 after
incubation for 5 days.
Method:
a) The percent dilution to be used must be determined. To
make this calculation, one should understand that
dilution water at room temperature contains approx-
imately 8 mg/1 of dissolved oxygen (DO). Consequently,
if the oxygen demand of the sample to be tested is
greater than 8 mg/1, dilution of the sample has to
be made. It is desirable to have at least 1 mg/1 of
initial oxygen left after 5-day incubation. Table 1
ia an aid to estimate the dilutions to use.
A-33
-------
Appendix 1
*Initial D0=7 mg/1
*Initial D0=8 mg/1
Percent
Dilution
(X)
1
2
3
4
5
6
7
8
9
10
15
20
25
50
Sample
added to 300-
ml. Bottle
(ml)
3
6
9
12
15
18
21
24
27
30
45
60
75
150
BOD Range
Min.
(mg/1)
210
105
70
53
42
35
30
26
24
21
14
11
8
4
Max.
(mg/1)
490
245
162
123
98
82
70
62
56
49
33
25
20
10
Percent
Dilution
(X)
1
2
3
4
5
6
7
8
9
10
15
20
25
50
Sample
added to 300-
ml. Bottle
(ml)
3
6
9
12
15
18
21
24
27
30
45
60
75
150
BOD Range
Min.
(mg/1)
240
120
80
60
48
40
34
30
27
24
16
12
9.6
4
Max.
(mg/1)
560
280
187
140
112
94
80
70
62
56
37
28
22
12
*Initial DO is the concentration of dissolved oxygen in mg/1 of the mixture of the
dilution water and the sample immediately after initial mixing.
Table 1. An Aid in Selection of Percent Dilution for BOD Determination
Raw sewage usually contains about 100 to 300 mg/1 BOD
so that 1- and 2-percent dilutions generally are used;
settled sewage BOD's usually range from 50 to 200 mg/1,
and 2- and 3-percent or 3- and 4-percent dilutions are
common; trickling filters use 5- and 10-percent; for
activiated sludge effluents, use 10-, 20-, or 50-per-
cent depending upon how good the effluent is. Very
strong sewages or industrial wastes are diluted 1 part
wastewater to 10 parts dilution water before making
the dilutions of 1- to 2-percent. In this way a range
of 1,000 to 3,000 mg/1 BOD is covered. However, the
inexperienced operator is advised not to try to
analyze industrial wastes.
A-34
-------
Appendix 1
b) Fill two 300 ml BOD bottles about half-full with dilu-
tion water.
c) Using a large-tipped pipette, measure the precalculated
amount of sample into the two 300 ml BOD bottles.
d) Fill each bottle with dilution water and insert stop-
pers the same way. See that all air bubbles are excluded.
e) Fill two additional bottles with straight dilution
water and insert stoppers the same way.
f) Incubate one bottle containing the diluted sample and
one bottle containing only dilution water.
g) Determine the initial DO levels of the diluted sample
and of the dilution water by running dissolved oxygen
determinations on the two remaining bottles.
h) After 5 days, run a dissolved oxygen determination on
the incubated bottles. Record the DO contents. (The
increase or decrease of DO in the bottles with just
dilution water is intended to serve only as a measure
of dilution water quality. There should be no increase
or decrease more than 0.5 mg/1 when compared to the
initial DO value of the dilution water.)
Calculations:
BOD values are calculated as follows:
100 x (Initial DO of diluted sample - DO of sample after 5 days)
Percent of sample added
= mg/1 (5 day BOD)
2) Dechlorination and Reseeding Procedure3
Whenever BOD determinations are to be made on chlorinated
water samples, sufficient reducing agent must be added to
remove the chlorine. After dechlorination, the sample must
be "reseeded" with organisms.
Method:
a) Secure an unchlorinated sample of raw sewage or primary
effluent about 24 hours prior to the time when you
expect to set up dechlorinated and seeded samples for
determination of BOD. Collect about one liter of
unchlorinated sample and let stand at room temperature
A-35
-------
Appendix 1
overnight. Pour off the clear portion of the sample
and use it for the "seed."
b) Check for the presence of chlorine in the composite
sample proceeding as follows:
(1) Carefully measure 100 ml of well-mixed sample
into a 250 ml Erlenmeyer flask.
(2) Add a few crystals of KI to the sample and dis-
solve the crystals.
(3) Add 1 ml of concentrated H^SO^ and mix well.
(4) Add five drops of starch.
1. If no blue color is produced and chlorine is
absent, the BOD of the composite may be deter-
mined without further treatment. In this
case, all of the chlorine has been "used up"
by the water and it may be assumed that a
sufficient number of organisms remains so
that the full BOD will be exerted.
2. If a blue color is produced, titrate the
composite sample with 0.025M NapS203-5H20 to
the end-point between the last trace of blue
color and a colorless solution. Make the
titration very slowly, counting the drops
of sodium thiosulfate used and recording the
number.
c) To dechlorinate a sample for BOD testing, measure out
another 100 ml portion of the well-mixed composite
into a clean 250 ml Erlenmeyer flask. Add the number
of drops of 0.025M sodium thiosulfate determined
necessary for dechlorination in step b4 above. Mix
well. Use this sample for determination of BOD. If
more sample is needed, place a larger sample into a
clean container and add a proportionate number of
drops of the sodium thiosulfate for dechlorinating.
d) For seeding of the sample, add 1 ml of the aged seed
(step a above) to each of the BOD bottles containing
dechlorinated sample.
e) Set up samples of the seed for determination of the
BOD using 2, 3, and 4 percent (3, 6, and 9 ml seed)
and determine the 5 day depletion due to 1 ml of seed.
A-36
-------
Appendix 1
Calculations:
If the sample has been dechlorinated and reseeded as
described, the 5 day BOD should be calculated as
follows:
B - (A + C)
0x 100 = 5 day BOD expressed as mg/1
where
A = 5 day DO depletion of seed sample/ml seed
B = Initial DO (mg/1) of diluted sample
C = DO (mg/1) of sample after 5 days
D = Percent of sample used
3) References
1. Water Pollution Control Federation, Simplified Labora-
tory Procedures for Wastewater Examination, WPCF
Publication, No. 18, Washington, D. C., 1968,
pp. 38-40.
2. Ibid., pp. 41-43.
5. Interpretation
Aided by natural selection, existing aquatic ecosystems have
evolved through geologic time. Organisms have adapted to their
environments to the extent that the components of these environ-
ments are now the very factors upon which they depend. Deviations
from this make-up, especially if sudden, may adversely affect the
organisms living there.
Even within a given locale, the environmental conditions which one
observes are limitless. Consequently, universal favorable concen-
trations of dissolved solids, gases, etc., are either exceedingly
difficult or impossible to identify. Since toxicity of chemicals
varies not only with the types and ages of the organisms concerned
but also with duration of exposure, temperature, accompanying dis-
solved and suspended substances, flow rate, etc., even generaliza-
tions concerning concentrations at which specific substances become
toxic are not feasible. Because of these difficulties, favorable,
tolerable and toxic concentrations are now indicated in this manual
on the premise that such information is, at its best, of little
significance or, at its worst, misleading.
The following activities are recommended as aids in the interpreta-
tion of chemical data (1).
A-37
-------
Appendix 1
a. Sample the ecosystem periodically over a long period of time.
Identify norms and note all biological and chemical changes,
especially those which occur suddenly. Evaluate your data in
terms of the entire ecosystem. Chemical determinations are
of limited significance alone.
b. Determine, in the laboratory, environmental factors which are
favorable or tolerable.
c. Use bioassay techniques to identify responses of organisms to
various concentrations of potential toxicants and try to deter-
mine permissible levels for the ecosystem under study.
d. Test the laboratory findings in the field to evaluate their
vali di ty.
To facilitate interpretation of test results, two tables are
included which emphasize those factors which are known to either
interfere with chemical tests (Table 1) or influence toxicity
(Table 2).
A-38
-------
Appendix 1
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Appendix 1
Footnotes: Table 1 .
1. Unhydrolyzed aluminum and/or iron (II) sulfate cause difficulty in
determining the end-point. Performance of the titration at boiling
temperature alleviates this problem.
2. Free chlorine may be removed by adding 1 drop 0.1M ^8203^20
to the titration sample.
3. Calcium carbonate and magnesium hydroxide precipitates cause fading
end-points and should be removed by filtration.
4. The presence of toxic substances such as heavy metals may interfere
with BOD determinations.
5. It also interferes in silver and mercuric nitrate tests.
6. It interferes in mercuric nitrate test.
7. It interferes in silver nitrate test.
8. pH must be in the range of 7-8. Errors are introduced above and
below this range.
9. The presence of algae may result in erroneously high Cl2 deter-
mination.
10. Temperature must be controlled at 20°C; otherwise, the C^ concen-
tration will vary.
11. Interference is caused by manganic manganese.
12. Color corrections may be made by using the orthotolidine-arsenite
method.
13. Azide modification of Winkler overcomes nitrite interferences.
Refer to Standard Methods for additional modifications.
14. pH values greater than 10 favor precipitation of CaC03, thus
causing drifting end-points which may yield low results (EDTA
Method).
15. Phosphates do not interfere in the tripyridine method for iron
determinations.
16. Periodate method for manganese determinations.
A-41
-------
Appendix 1
MODE OF INTERFERENCE Table 1.
a. 0 interferes with reaction mechanism.
b. X interferes with phylometric readings.
c. - interferes with end-point determination.
d. Test does not differentiate.
e. It forms interfering ppt under conditions of test.
f. It disturbs carbon dioxide - carbonate equilibrium.
g. It alters reaction rate.
h. It alters concentration.
* The material for this table was obtained from Standard Methods for the
Examination of Water and Wastewater. This reference should be consulted
for further information.
A-42
-------
Appendix 1
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A-43
-------
Appendix 1
6. Bibliography
a. American Public Health Association, Standard Methods for the
Examination of Water and Wastewater, (13th ed.), American
Public Health Association, Inc., New York City, 1971. This
includes detailed information concerning the identity,
origin, and analysis of numerous chemical parameters. It
is an indispensable reference for water pollution studies.
b. Federal Water Pollution Control Administration, Report of the
Committee on Water Quality Criteria, Superintendent of
Documents, U. S. Government Printing Office, Washington,
D. C., 1968. This is a compilation of FWPCA water quality
criteria recommendations and supporting information.
c. McKee, J. E.,and H. W. Wolf, Water Quality Criteria, (2nd ed.),
Water Quality Control Board, Sacramento, Calif., 1963.
Although dated, this is an outstanding compilation of the
technical, social, and legal history of water quality
criteria. Reviews of Federal and State policies as well
as commentaries on an enormous number of chemical pollutants
are included. 3827 references are cited.
d. Sawyer, C. N.,and P. L. McCarty, Chemistry for Sanitary
Engineers, (2nd ed.), McGraw-Hill Book Co., New York City,
1968. This is a lucid presentation of the theory and
methods of sanitation chemistry intended for the reader
possessing a solid foundation in elementary chemistry.
A-44
-------
Appendix 1
B. Bacteriology
There are numerous types of bacteria and most environments are
capable of supporting some bacterial life. Certain bacteria, in-
cluding those used in food processing or those found in the soils
which enable plants to obtain nutrients, are beneficial. Some bac-
teria, however, cause food to decay and are responsible for various
diseases.
Water can contain many types of bacteria in large numbers Some
of these bacteria are harmless to man, but certain types, termed
pathogenic, cause diseases such as typhoid fever, dysentary, and
cholera. The possible danger of disease dictates that water be
free of pathogenic bacteria. Because the cultivation of patho-
genic forms is difficult and requires trained personnel, a group
of more easily cultivated bacteria is used to indicate the pos-
sible presence of pathogens. These indicator organisms include
total coliform, fecal coliform, and fecal streptococci organisms.
1. Total Coliforms
Total coliforms include a group of rod or stick-shaped organ-
isms characterized by their ability to ferment a specific
sugar (lactose) at 35°C within 48 hours. Although coliforms
are introduced to water supplies via water runoff from soil,
drains, etc., they are considered significant as indicator
organisms because of their predominance in the intestinal
tracts of warm-blooded animals. While not all animal wastes
contain pathogens, the excrement of diseased animals and
animals serving as carriers of pathogens do present health
hazards.
The total coliform density is roughly proportional to the
amount of excremental waste present. With exceptions,
elevated coliform populations are suggestive of significant
contamination by excrement of warm-blooded animals. Several
factors which cause fluctuations in total coliform popula-
tions are summarized in Table B-l.
Table B-l Factors Influencing Total Coliform Levels
Higher Lower
1. Sewage intrusion 1. pH changes
2. Nutritive effluents (contain- 2. Temperature changes
ing sugar, dairy wastes, etc.)
3. Storm drain overflows 3. Land runoff (pro-
longed rain)
4. Land runoff (initial storms) 4. Toxic wastes
A-45
-------
Appendix 1
Coliform population limits have been set by federal and state
health services. These limits vary with the designated use
of the water supply and are quite variable within such desig-
nations.
Table B-2
Water Quality Criteria with Respect to Total Coliform Popula-
tions
Class of Water
Mass. N.H. Vt.
a. Drinking water 1 * 1 * 1 *
b. Water of highest quality,
designated for ingestion after
disinfecting supplies. 50 50 50
c. Suitable for bathing and
recreation; irrigation and
agricultural uses, good fish
habitat; good aesthetic value;
acceptable for ingestion fil-
tration and disinfection. 1,000 240 1,000
d. Suitable for recreational
boating; irrigation of crops
not consumed raw; habitat for
wild life and fish; certain
industrial cooling and process-
ing use. Unspecified
e. Suitable for aesthetic enjoy-
ment, power, navigation, and
certain industrial cooling and
processing uses. Unspecified
*This is a national standard specified by U.S.P.H.S.(1962)
Fecal Coliform
Fecal coliform, a component of the total coliform population,
is characterized by its ability to reproduce on a special
medium (M-FC) at a temperature of 44.5 to 5QOC. Because non-
fecal coliforms may grow below 44°C and fewer fecal coliforms
grow above 45°C, temperature maintenance within the specified
tolerance is critical.
A-46
-------
Appendix 1
Fecal coliforrns are gaining notoriety as pollution indices
because of their relatively infrequent occurrence except in
association with fecal pollution. Moreover, because survival
of the fecal coliform group is shorter in environmental water
than for the coliform group as a whole, high fecal coliform
levels indicate relatively recent pollution.
When accompanied by fecal streptococci counts, fecal coli-
form values may aid in the differentiation of animal from
human waste (Table B-3). However, caution must be exercised
in the interpretation of such data because of technical dif-
ficulties in performing precise counts.
Table B-3
Average Individual Density Per Gram of Feces (Indicator Micro-
organisms From Some Animals)
Animal
Man
Duck
Sheep
Chicken
Cow
Turkey
Pig
Fecal
Col iform
Million
13.0
33.0
16.0
1.3
0.23
0.29
3.3
Fecal
Streptococci
Million
3.0
54.0
38.0
3.4
1.3
2.8
84.0
Ratio
Fecal Col ./Fecal Strep.
4.4
0.4
0.4
0.4
0.2
0.1
0.04
It is anticipated that national standards for water use will
be established according to population densities of fecal
coliform as shown in Table B-4.
Table 6-4
Water Quality Criteria With Respect to Fecal Coliform Population
Kind of Water Recommended Numbers
of Fecal Coliforms
1. Water designated for Primary 1. Should not exceed a
Contact Recreation mean of 200/100 ml
2. Water other than for Primary 2. Should not exceed a
Contact Recreation mean of 1000/100 ml
3. General Recreational Surface 3. Average not to exceed
Water 2000/100 ml
A-47
-------
Appendix 1
Primary Contact Recreational Activities are defined as
those activities in which there is a prolonged and intimate
contact with the water involving considerable risk of in-
gesting water quantities sufficient to pose a significant
health hazard.
The Food and Drug Administration recommends that the surface
waters above shell fish beds shall not have fecal coliform
counts above 70/100 ml.
3. Fecal Streptococci
Fecal streptococcus, as used in this discussion, refers to
any streptococcus commonly found in significant numbers in
the feces of human or other warm-blooded animals. Fecal
streptococci are spherical organisms which generally occur
in pairs or short chains when viewed microscopically. They
are capable of reproducing at 45°C and, in some instances,
at 10°C on a selective medium containing sodium azide and
other inhibitors.
Because fecal streptococci do not occur in pure water or
virgin soil, their presence in water supplies indicates the
existence of warm-blooded animal pollution. Their validity
as an index of pollution is enhanced by their inability to
reproduce in water supplies. Moreover, fecal streptococci
are resistant to salts; therefore, this group could have
special value for salt water investigations.
Fecal streptococci determinations, when accompanied by fecal
coliform studies, serve as a valuable tool in the differen-
tiation of animal from human wastes (Section 2, Table B-3).
In intestinal wastes of human origin, the ratio of number of
fecal coliforms to number of fecal streptococci tends to be
greater than four. In comparison, when such ratios are de-
termined for intestinal wastes from nonhuman sources;, the
values tend to be markedly less than 0.7. When interpreting
fecal streptococci data, the following three points should
be considered.
a. The presence of fecal streptococci in untreated
water indicates the presence of fecal pollution
by warm-blooded animals.
b. In samples where the source and significance of
the coliform group have been questioned, the
presence of the streptococcus group should be in-
terpreted as indicating that at least a portion
of the coliform group is derived from fecal
sources.
A-48
-------
Appendix 1
c. Because of the uncertainties in die-off rates,
the absence of fecal streptococci does not
necessarily mean that water is bacteriologically
safe.
4. General Procedures
a. Sterilization
It is necessary to sterilize all necessary equip-
ment in order to assure that only bacteria from
the collected water sample will be counted. There-
fore, the following instructions must be followed
closely.
Equipment:
autoclave or pressure cooker
Items for immediate sterilization;
1) for preparation of media (Procedure b_)
Petri dishes (number determined by sampling
needs)
2) for sample dilution (Procedure ej
1 100-ml graduated cylinder
3 1-ml pipettes
3 125-ml pipettes
3 125-ml flasks
distilled water
3) for filtration (Procedure f)
1-, 5-, 10-ml pipettes (number determined by
sampling needs)
Items to be sterilized as needed:
1) for preparation of media (Procedures b_ and e)
collection of bottles containing 0.2 ml 10%
sodium thiosulfate
A-49
-------
Appendix 1
2) for preparation of solutions (Procedures c_ and f_)
phosphate buffer in closed container
Method:
1) All glassware and distilled water should be auto-
claved. Use 15 Ibs. of pressure at 121°C for 15
minutes. The openings of all flasks and cylinders
should be wrapped with aluminum foil. Do not auto-
clave plastic parts or culture media without con-
sulting the manufacturer's instructions.
2) Plastic parts can usually be sterilized by boiling
in a water bath for 3 minutes.
3) Filter membranes come presterilized. However, after
use, they may be washed off in 95% ethanol, auto-
claved at 12 Ibs. for 12 minutes for reuse.
Note: Reuse membranes for the same media only.
4) Petri dishes (Millipore) are presterilized. For
reuse, they should be soaked in liquid household
bleach for 10 minutes and rinsed thoroughly under
running water. Then they should be immersed in 70%
isopropyl alcohol for 10 minutes and dried. Fol-
lowing assembly, they may be stored for later use.
b. Preparation of Media
After bacteria are collected and filtered from the water
sample, they must be allowed to grow at precise temper-
atures into visible colonies which can be counted easily.
To facilitate growth, the proper nourishment (culture
medium) must be provided. Total coliform, fecal coliform,
and fecal streptococci require different types of culture
media, prepared as follows:
Equipment: (Note glassware need not be sterile.)
3 125-ml flasks
3 100-ml graduated cylinders
2 2-ml pipettes
2 glass stirring rods
1 balance
1 heat source
A-50
-------
Appendix 1
Reagents and Media:
Dehydrated M-Coliform Broth (MF Endo Broth)
Dehydrated M-FC Broth Base
Dehydrated M-Enterococcus Agar
Agar
Ethanol (95%)
Rosalie Acid
Sodium Hydroxide
Distilled Water (300 ml)
Procedure:
The following methods describe the preparation of agar
media for use without pads. This approach is more con-
venient for preparation of multiple plates. If absorption
pads and broth are preferred, omit the agar and add 2 ml
of broth to each pad.
1) Total coliform using M-Coliform or MF Endo broth base
a) Pipette 2 ml 95% ethanol into a 100-ml graduated
cylinder and fill with distilled water to the
100 ml mark. Transfer this to a 125-ml flask.
b) Add 4.8 g of dehydrated M-Coliform or MF Endo
broth base and 1.5 g of agar to the diluted
alcohol solution. Mix thoroughly. Note: MF
Endo broth base can be purchased with agar al-
ready added. If this is done, follow the manu-
facturer's instructions for preparation.
c) Cover flask with a foil cap and heat the mixture
with agitation until it just begins to boil. (Do
not reheat or prolong the heating. This reduces
the selectivity of the media.)
d) Cover the bottom of each sterile petri dish with
the broth. This should be done while the broth
is still warm. It will gel as it cools, allowing
the filter membrane to be placed directly on it.
Three or 4 dishes are normally prepared for each
A-51
-------
Appendix 1
water sample (refer to Section e). Petri dishes
prepared in this way may be refrigerated for 24 hours.
2) Fecal coliforms using M-FC Broth base
a) Dissolve 0.8 g NaOH in 50 ml distilled water
b) Dissolve 0.1 g rosolic acid in 10 ml 0.2M NaOH
prepared in Step a_.
c) Place 3.7 g M-FC Broth and 1.5 g agar into a
125-ml flask.
d) Pipette 1 ml 1% rosolic acid solution (prepared
in (b)) into a 100-ml graduated cylinder and
add distilled water to the 100 ml mark. Pour
this mixture into the flask containing the agar
and broth base.
e) Place a foil cap on the flask and heat, with con-
tinuous agitation, to the boiling point. Remove
from the heat immediately to avoid destruction of
the selectivity of the medium.
f) Cover the bottom of each sterile petri dish with
the warm medium. Dishes prepared according to
these instructions may be refrigerated for one
week.
3) Fecal streptococcus using M-Enterococcus agar
a) Add 4.2 g M-Enterococcus agar to 100 ml distilled
water.
b) Heat to boiling. Remove from the heat immediately
to avoid destruction of the selectivity of the
medium.
c) Cover the bottom of each sterile petri dish with
the warm medium. The prepared dishes may be
stored in a cool, dark place for 1 week.
References:
(1) Microbiological Analysis of Water, Mi Hi pore
Corp., Bedford, Mass., 1969, pp. 3 and 5.
(2) Mi Hi pore Experiments in Microbjology, Millipore
Corp., Bedford, Mass., 1969, pp. 17-19.
(3) U. S. Department of the Interior, Current Practices
in Mater Microbiology, U. S. Government Printing
Office, Washington, D.C. ,1969.
A-52
-------
Appendix 1
c. Preparation of Solutions
1) Sodium thiosulfate
Chlorine is added to public water supplies and
sewage treatment effluents to kill bacteria. When
collecting water samples, it is necessary to "de-
activate" any chlorine present to avoid killing
bacteria after they are trapped in the collection
bottle. If this is not done, false data suggesting
low or absent quantities of bacteria may be obtained.
Sodium thiosulfate is used to "deactivate" the chlo-
rine and may be prepared and stored. When needed,
the correct amount of sodium thiosulfate is added
to the collection bottle and is autoclaved to assure
sterilization.
Equipment:
1 1-ml pipette
1 125-ml flask
250-ml collection bottles (number determined by sam-
pling needs)
metal foil or paper
Reagents:
Sodium Thiosulfate
Distillled Water
Procedure:
a) Dissolve 10 g NazSgOa'SHzO in 50 ml distilled
water and dilute to 100 ml.
b) Add 0.2 ml of this sodium thiosulfate solution
to each 250-ml bottle. If glass stoppered bot-
tles are used, place a thin strip of paper in
the neck to avoid "freezing" of the stopper.
c) Seal cap and neck with foil and autoclave using
15 Ibs. pressure at 121°C for 15 minutes. After
the bottles are sterilized, they should remain
sealed until the time of collection. Label each
bottle to identify it as sterilized and ready for
use.
A-53
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Appendix 1
2) Phosphate buffer
The collected water samples are to be poured through
a membrane filter to trap the bacteria in a later
step. It is necessary to rinse the funnel with a
sterile phosphate buffer solution to assure that all
bacteria are washed onto the membrane. The phosphate
buffer may be prepared at any time and sterilized
when needed for use.
Equipment:
1 500-ml flask
1 1-liter flask
1 1-liter glass bottle (preferably with glass
stopper)
2 2-ml pipettes
pH meter or close range pH paper
Reagents:
Potassium Dihydrogen Phosphate (KH2.P04)
Sodi urn Hydroxi de
Distilled Water
Method:
a) Dissolve 34.0 g KH2P04 in 250 ml distilled water
and dilute to 500 ml.
b) Prepare a 1M NaOH solution by dissolving 4 g
NaOH in 50 ml of distilled water and diluting
to 100 ml.
c) Add the 1M NaOH drop by drop to the solution of
KH^P04 (Step a) until the pH is 7.2. Read the pH
using a pH meter.
d) Dilute the adjusted solution to 1 liter with dis-
tilled water. Label the solution as "Stock
Phosphate" and store for later use.
e) Add 1.25 ml of this phosphate solution to each liter
of distilled water being converted to the "working"
phosphate buffered water.
A-54
-------
Appendix 1
f) Autoclave the buffered water in a closed glass
bottle using 15 Ibs. pressure at 121°C for at
least 15 minutes. Be sure to allow the autoclave
pressure to drop slowly!
d. Collection of Water Sample
When collecting a water sample, it is important to select a
location which is representative of the body of water. The
collection of water too close to the shore or within stag-
nant areas may not yield a representative sample. A thorough
analysis involves sample collection at varying depths (i.e.,
about every three feet). However, analysis for recreation
usage requires only that a sample be taken one foot under the
surface in the middle of the swimming area. For larger streams
or rivers, a sterile device can be lowered from a bridge into
the main current and filled. The sample must not be contam-
inated by surface scum or unnatural turbidity at any time
during or after the collection.
Equipment:
250-ml sterilized collection bottles containing 0.2 ml
sodium thiosulfate solution (added before sterilization)
Method:
1) Remove the foil hood and paper strip from the stopper
of the sterile bottle.
2) Place the entire bottle under the water in an inverted
position and turn it upright. Keep your hands clear of
the water entering the bottle. In moving water, it is
wise to keep your hands downstream relative to the neck.
3) Fill the bottle about 2/3 full and replace the stopper
while the entire unit is still submerged. The air space
is left in the bottle to allow adequate mixing of the
sample later. The sample should be iced immediately
after collection. Samples may be held a maximum of 6
hours in the field if necessary, and an additional 2
hours in the laboratory.
e. Filtration Volumes - Selection and Dilution
In using membrane filtration as a means of detecting bacteria
in water, definite limitations arise concerning the number of
countable bacteria colonies on each membrane filter. If too
many colonies develop, some may be fused together making ac-
curate counting impossible. If there are not enough, the
count may not be representative. Therefore, certain ranges
A-55
-------
Appendix 1
of colonies on each membrane are used as criteria for se-
lecting the dishes which will give the most accurate repre-
sentation of the bacterial density population (Table B-5).
Table B-5 Recommended Ranges of Colony Counts for Membrane
Filtration
Techniques
Test Number of Colonies
Minimum Maximum
Total coliform 20 80
Fecal coliform 20 60
Fecal streptococci 20 100
Total bacteria counts 20 200
The selection of sample volumes which result in counts within
the above ranges depends upon the actual bacterial population
of the sample. A summary of relationships which exist between
filtration volume bacterial levels within samples is presented
in Table B-6.
Table B-6 Ranges Covered By Representative Filtration
Volumes
ml sample
filtered
100
10
1
0.1
0.01
Bacterial count
20 colonies
20
200
2000
20,000
200,000
60 colonies
60
600
6000
60,000
600,000
per 100 ml based on
80 colonies
80
800
8000
80,000
800,000
100 colonies
100
1000
10,000
100,000
1,000,000
A-56
-------
Appendix 1
As indicated in the Table, a moderately polluted lake con-
taining several thousand organisms/100 ml would require
the filtration of at least 1 ml undiluted water.
1) Procedure for selection of filtration volumes
A direct calculation of the filtration volume can be
made if there is prior knowledge of the bacterial pop-
ulation density in the water under study. The follow-
ing relationship is utilized:
Sample filtration A
= 100 X
volume (ml) average count/100 ml
where A = mid-range number of colonies for an acceptable
plate count which varies according to the or-
ganisms being detected as follows:
A = 50 for total coliform counts
A =40 for fecal coliform counts
A = 60 for fecal streptococci counts
As a sample problem consider a stream with an estimated
total coliform level of 25,000/100 ml. The calculation
is: 100 X 50
25,000 total coliform/100 ml
thus giving a required filtration volume of 0.2 ml.
To avoid disappointing results, 3 or 4 different volumes
should be analyzed to increase the likelihood that at
least one membrane will possess an acceptable number of
colonies. The following guidelines may aid in the selec-
tion of these varying filtration volumes.
a) Total coliform counts should be based on filtration
volumes varying by a factor of 4 or less.
b) Fecal coliform counts should be based on filtration
volumes varying by a factor of 3 or less.
c) Fecal streptococci counts should be based on filtra-
tion volumes varying by a factor of 5 or less.
If no prior bacterial data are available for a body of
water under investigation, Table B-7 will be of assist-
ance.
A-57
-------
Appendix 1
Table B-7
Filtration Volumes for Waters Not Previously
Studied
Filtration Volumes (ml) *
Source Total Coliform Fecal Coliform Fecal Strep.
Unpolluted Raw
Surface Water
1,4,15,60
(33-8000)
1,3,10,30
(67-6000)
(20-10,000)
Polluted Raw .02,.08,.15, .5
Surface Water (4,000-400,000)
.1,. 3, 1.0, 3.0
(670-60,000)
0.1,0.5,2.0 for
Animal Pollution
1000-100.000
(0.2,1.0,5.0) or
(400-50,000)
Sewage and
.0003,.001,.003
Dilute Sewage (200,000-
27,000,000)
.003,.001,.003
(670,000-
0.2x,lx.,5x Total
Colif. for Animal
Pollution
20,000,000) lx,5x,25x Total
Colif. otherwise
* Ranges/100 ml covered by the recommended volumes are enclosed in
parenthesis.
2) Procedures for dilution
NOTE: Dilution is necessary only if an acceptable count
(see Table B-5) was not obtained by using different fi 1 -_
tration volumes of the undiluted raw sample. If dilution
is necessary, follow the procedures below, being sure to
filter the total diluted volume.
The following procedure is for the preparation of a
1:1,000,000 dilution.
To choose the correct dilution, it is convenient to change
the filtration volume to scientific notation involving a
volume that can be easily pipetted. For example., 0.002 ml
can be written as 0.2 ml x 10"2. Then 0.2 ml of a 1:100
(or 10~2) dilution can be pipetted for filtration.
Equipment:
3 1 ml pipettes (sterilized)
1 100 ml graduated cylinder (sterilized)
3 125 ml flasks (sterilized)
distilled water (sterilized)
A-58
-------
Appendix 1
Method:
a) Place 99 ml sterile distilled water into each of
three sterile flasks.
b) Shake the sample bottle thoroughly to assure a uni-
form distribution of bacteria.
c) Add 1 ml sample water to Flask 1 containing 99 ml
sterile distilled water. Mix well. This gives a
dilution of 1:100.
ml of mixture from Flask 1 and place into
Mix well. This gives a 1:10,000 dilution
d) Remove 1 ml
Flask 2. Mix well. This gives a 1:10,000 dilution,
e) Remove 1 ml of mixture from Flask 2 and place into
Flask 3. Mix well. This gives a 1:1,000,000 dilu-
ti on.
(For higher dilutions, continue this procedure.)
f. Preparation of Filter for Incubation
Now that the glassware is prepared, the sample collected and
diluted, and the petri dishes filled with medium, you are
ready to catch and to grow the bacteria. When the water sam-
ple is passed through the membrane filter in the filtration
apparatus, the bacteria in the water are trapped on the filter.
By removing the filter and placing it in a petri dish prepared
with appropriate nutrients, the bacteria are ready for growth
in the incubator. Because of the need to transfer the filter
from the filtration apparatus to the petri dish, it is essen-
tial to follow the aseptic techniques outlined below.
Equipment:
filtration apparatus *
vacuum system (syringe, hand pump and fittings, or
standard vacuum pump)
forceps without corrugations on the inside of the
tips
methanol or ethanol
gas burner or alcohol lamp
large, wide container for boiling water bath (if
filtration apparatus is not presterilized)
A-59
-------
Appendix 1
stand to support (lined) flask over burner
sterilized petri dishes containing nutrients
flasks with diluted samples of water being tested
25-ml graduated cylinder (sterilized) - one for
each different water sample
presterilized membrane filters
sponge or cloth
100 ml beaker for soaking forceps
sterilized pipettes (number and capacity is deter-
mined by sampling needs)
* This procedure assumes apparatus to be unsterile, but
autoclaving in advance is acceptable for Mi Hi pore
Sterifil Filtration System.
Filtration
Funnel
(Threaded Bottom)
Membrane
Gridded Filter
Filter Support
(Threaded top) ./? /
Filter Base -/
-------
Appendix 1
Method:
1) Set up a boiling water bath by placing a large,
wide-mouthed container, containing distilled water,
on a stand above a gas burner.
2) Disinfect a laboratory table surface by swabbing it
with alcohol. Allow the surface to dry before pro-
ceeding. Also swab the bottoms of apparatus to be
placed on the table.
3) On the work surface, assemble the funnel unit and
receiver flask and connect it to the vacuum system.
(Fig. B-l)
4) Set out the petri dishes (raised-lettered side down)
and collection bottles.
5) Pour about 20 ml of methanol or ethanol into a small
flask and place forceps into the solution. Before
using the forceps, they should always be placed brief-
ly in a direct flame from a burner or alcohol lamp
to burn off the excess alcohol. Allow them to cool
slightly before touching the highly flammable membrane
filter.
6) Immerse the filtration funnel and base in the boiling
water bath for 3 minutes. After removing them from
the water, attach them to the receiver flask. Loosen
the funnel to allow placement of the membrane filter.
7) Using flamed forceps, remove a sterile membrane from
a sealed package. Discard the blue wax packaging
disk and reseal the filter envelope. Place the mem-
brane, grid side up, over the porous plate of the
filter support on the filtration apparatus. Carefully
place the funnel over the filter base and lock it in
place.
8) Pour 20 to 30 ml of sterile buffered water into the
funnel. Check for leakage around the funnel base.
(If any occurs, repeat Steps 7 and 8.) Leave the
buffer solution in the funnel.
9) Shake the prepared sample thoroughly and, using the
highest dilution first, pipette the predetermined vol-
ume (Procedure e) into the buffer solution in the
funnel. (Be sure the pipette is sterilized.) If more
than 20 ml of the sample is to be filtered, a sterile
graduated cylinder may be used in lieu of the pipette.
A-61
-------
Appendix 1
10) Reduce the pressure in the receiver flask by creating a
partial vacuum in it. Use either a hand pump (e.g.,
Millipore Vacuum System) or an electric pump.
11) Rinse the funnel by filtering three volumes of 20 to
30 ml of sterile buffered water through the membrane.
12) Loosen the filtration funnel. Remove the cover of a
petri dish. Remove the filter membrane from the fil-
ter support with flamed forceps. Place the filter
membrane, grid side up, on the medium in the petri dish,
using a rolling motion to avoid trapping any air.
Replace the lid on the dish and label it on the bottom,
identifying the sample and the filtration volume.
13) Before preparing the next membrane, sterilize the forceps
by putting them into the alcohol. Do not forget to
flame them. (The rinse with the buffer solution is
sufficient to clean the funnel.)
14) Starting with Step 7, repeat the procedures for the re-
maining sample volumes.
Note: The buffer rinses (Step 11) clean the funnel suf-
ficiently for all subsequent filtration unless analyzing
water samples for drinking purposes.
Incubation
As pointed out in "Preparation of Media," bacteria need cul-
ture media to grow well. The bacteria also need warmth, mois-
ture, and darkness to grow rapidly. Therefore, the petri
dishes containing the bacteria are placed in an incubator to
assure proper conditions.
Method:
1) Incubation of total coliforms
After preparation, invert the petri dishes con-
taining total coliform cultures and place them
in a standard incubator for 24 hours at 35-
0.5°C.
Note: The humidity within the incubator must approach
100?^; however, when tightly sealed plastic petri dishes
are used, a portion of the broth evaporates, raising
the humidity within the dish itself to 100% and making
A-62
-------
Appendix 1
adjustment of the humidity outside the dishes unnecessary.
If dish covers are loose fitting, the humidity can be
maintained by placing a vegetable crisper containing wet
towels in the incubator. The dishes are then placed on
top of the towels and covered with the crisper's lid.
2) Incubating fecal coliform cultures
a) After preparation, invert the petri dishes containing
fecal coliform cultures and place them in waterproof
plastic bags (3 to 6 dishes per bag).
b) Submerge the bags in waterbath incubator and incubate
at 44.5±0.2°C.
Note: The temperature of the waterbath is critical. Above
44.7°C. counts drop rapidly. Below 44.3°C specificity is
lost. Therefore, no more than 20 minutes should elapse
between filtration and incubation to prevent nonfecal coli-
form colonies from developing at lower temperatures. Sub-
mergence in waterproof plastic bags reduces the temperature
equilibrium time considerably.
3) Incubating fecal streptococcus cultures
After innoculation, invert the petri dishes containing
fecal streptococcus cultures. Place them in a standard
incubator for 48 hours at 35*0.5°C.
h. Counting Techniques
If all the previous steps were carefully followed, bacterial
colonies should now be visible for counting. The results of
the count allow us to determine whether the water source is
polluted and, if so, how badly.
Method:
1) Counting
a) Place the petri dish to be counted under the micro-
scope after removing the lid. Lighting for the counts
must be from a fluorescent light source as close to
directly above the petri dish as possible; the image
of the light source is reflected off the colony sur-
faces into the microscope.
b) Count colonies in an orderly back-and-forth sweep
from top to bottom of the filter, using grids as
channels. Be sure to avoid mixing any colonies or
counting any colonies twice simply because they are
A-63
-------
Appendix 1
in contact with a grid-line. Count all colonies
individually. Even if two or more are in contact,
almost invariably, they show a fine line of contact.
Other individual colonies may have grown to unusual
shapes because of particles or fibres that may have
found their way onto the filter membrane. A hand
tally is convenient for counting the colonies.
Note: In dishes of total coliform colonies, the
coliform colonies demonstrate a greenish metallic
luster, or "sheen," which may cover the entire sur-
face of the colony or may appear only in the center
of the colony. (Any amount of "sheen" production
denotes a coliform colony.) Noncoliform colonies
are lighter and do not show this "sheen" even though
they may be shiny. In the fecal coliform dishes, the
fecal coliform colonies are blue and all other col-
onies are cream colored (any amount of blue is posi-
tive). Fecal streptococcus colonies have a reddish
hue, while other colonies range from cream to clear.
2) Calculations:
The count from your membrane must be adjusted for the
dilution of the sample and the volume filtered. Results
should be in coliforms per 100 ml
Number of Coliform/100 ml
= Number of colonies
Vol. of filtration sample x 10°
For your calculations use the petri dish from
the dilution that gives a direct count between
20 and 80. If there are too many colonies on
all three dishes, the number is recorded as
"T. N. T. C." (too numerous to count). The fol-
lowing shows sample data and calculation:
Dish #1 Dish #2 Dish #3
No. of coliform TNTC 39 7
Dilution 1:100 1:10,000 1:1,000,000
Volume filtered 20 ml 20 ml 20 ml
Dish #2 is used for the calculation because
the number of colonies was between 20 and 80.
Number of coliform/100 ml
= 39 X 100 = 1,950,000/100 ml
20x 1/10,000
A-64
-------
Appendix 1
i. Disposal of Cultures
Cultures, whether on filters or in any other medium should
be handled with the utmost care, as if they were all poten-
tially dangerous. When you have completed the experiment
and observed the results, the cultures should be destroyed
or deactivated, and the the petri dishes resterilized. This
can be accomplished by the following procedure:
1) Using forceps, carefully remove the petri dish covers
and place both covers and dishes into a large beaker
or pan containing liquid household bleach for 10 min-
utes. Unless membranes are to be preserved or reused,
they should also be soaked in the bleach.
2) If membranes are to be reused, they should be soaked
in ethanol to destroy the colonies. Then sterilization
should be completed according to page A-50, step 3.
3) Membranes may be preserved with colonies by drying on
a paper towel. They may even be reconstituted with
distilled water for later demonstrations.
5. Bibliography
a. American Public Health Association, Standard Methods for the
Examination of Water and Wastewater, (13th ed.), American
Public Health Association, Inc., New York City, 1971.
This is an essential, comprehensive reference for water
quality studies, which includes methods of qualitative
and quantitative bacterial investigations.
b. Federal Water Pollution Control Administration, Report of the
Committee on Water Quality Criteria, U. S. Government
Printing Office, Washington, D. C., 1968. It summarizes
FWPCA recommendations for water classification categories
and criteria and serves as a valuable aid in the inter-
pretation of test results.
c. McKee, J. E., and H. W. Wolf, Water Quality Criteria, (2nd
ed.), Water Quality Control Board, Sacramento, Calif.,
1963. This is a thoroughly documented coverage of nation-
wide water quality policies, biological effects of pol-
lutants, and judicial action.
d. Pelczar, M. J., and R. D. Reid, Microbiology, McGraw-Hill
Book Co., New York City, 1965. This elementary micro-
biology textbook includes introductions to the taxonomy,
biochemistry, cultivation, control and ecological roles
of microorganisms.
A-65
-------
Appendix 1
e. Microbiological Analysis of Hater, Mi Hi pore Corp., Bedford,
Mass., 1969.A variety of specific techniques for the
isolation and identification of bacteria in water samples
is presented.
f. Experiments in Microbiology, Mi Hi pore Corp., Bedford, Mass.,
1969. It contains an illustrated introduction to mem-
brane filtration and culturing techniques and theory
and is supplemented by experiments oriented toward the
beginning student.
A-66
-------
Appendix I
C. Aquatic Biology
A biological investigation of an aquatic community should lead to
an understanding of the extent to which it has been affected by
man. In order to obtain this understanding, those doing the
investigating must be able to assess the biological effects of
pollution, to identify organisms, to understand and employ field
and laboratory procedures, and to interpret the data which they
collect. The following section contains information which will
be useful to those conducting a biological investigation.
1. The Basis of the Biological Evaluation of Pollution
Pollutants may affect aquatic environments in two ways.
Indirectly, they may produce modifications such as altering
the food chain, changing the average annual temperature, or
reducing the concentration of dissolved oxygen. Directly,
pollutants may act physically or physiologically on the
resident organisms.
Biological effects of pollutants may be studied by field
observation, laboratory evaluation, or both. In the field,
evaluations are usually based on comparisons with an actual
or imaginary unpolluted reference or "control" site. For
example, the aquatic life downstream from a point of pollution
might be compared with that upstream from the source of
pollution. Also, the quality of aquatic life in a polluted
area may be compared to previous conditions in that area if
"prepollution" studies had been made.
Likewise, laboratory evaluation almost always involves a
comparison, or control, setup. For instance, the bioassay
technique involves the exposing of some type of organism to
a series of concentrations of some substance for a stated
period of time under controlled conditions.
Qualitative and quantitative evaluations may be used to
indicate the "health" of aquatic environments. The indicator
concept is based on the idea that there must be some organism
which is found only in polluted areas. This is true only for
certain types of bacteria which are present in the intestines
of warm-blooded aminals. Finding these bacteria demonstrates
a strong likelihood that animal excrement is present. Higher
forms of life which are pollution-tolerant may also be found in
clean water; therefore, finding of these organisms does not
necessarily indicate pollution. The species composition of
an aquatic community is sensitive to environmental conditions
and hence to pollution.
A-67
-------
Appendix I
Quantitative data may refer to an entire aquatic community or
only to selected or individual taxon. Numbers of individuals,
quantity (biomass), or both may be investigated. Rate of
production, food web interrelationships, and energy flow might
also be investigated.
2. The Identification of Aquatic Organisms
Questions usually posed about an organism which is seen for the
first time are "What is it?" and "What is its name?" Because
there are over a million and a half kinds (species) of
organisms known, a rather elaborate file reference system must
be used for naming and classification.
The system of biological nomenclature consists of a series of
groupings, or taxa (singular: taxon). Species is the founda-
tion taxon. Similar species are grouped into genera (singular:
genus), and similar genera are grouped into families. The
system continues in a like manner to order, class, phylum
(plural: phyla), and kingdom. Kingdom is thus the broadest, or
most inclusive, taxon.
An investigator must make a decision, based upon the purposes
for which he wishes to use his data, as to how precisely he
wishes to identify the organisms which he observes. For some
purposes he might want to know only how many kinds of organisms
were present. In this case he could simply designate them as
species (a), species (b), species (c), etc. These designations
would be based upon careful observations of likenesses and
differences of the organisms studied. This type of
"classification" would be acceptable for beginning students.
If more precision is desired, an investigator would probably make
use of an identification key.
The following key to basic types of plankton and small
aquatic organisms should enable the student to determine the
general type of organism he is observing. He may then
wish to proceed to a more complete key. Identification to
species level will usually be very difficult and should only
be attempted under the guidance of a taxonomist of recognized
competence in the particular taxon in question.
A-68
-------
Appendix I
Key To Types of Plankton And Other Aquatic Organisms*
Read each question in turn, refer to the specimen for the answer.
If the answer is yes, proceed to the paragraph indicated in the
"yes" column; if no, turn to the paragraph number in the "no"
column. If you have made a mistake, the "no" column reference
may send you back to reexamine some earlier decision.
If the answer is "yes," there may be no number given in the "yes"
column, but there will be a name in capital letters which is the
name of the group of organisms to which the specimen belongs, and
a plate number is cited which illustrates one or more examples.
*This Key was prepared by Dr. H. W. Jackson,
Training Program, Federal Water Pollution
Control Administration, Cincinnati, Ohio.
Yes No
1. Is it necessary to use a microscope to see
the organism? 3 2
2. Is the organism, or a mass of it, some
shade of green or brown? (The shape is
probably stringy, round or shapeless.) 9 13
3. Is the body relatively complex, with
many tiny active hairs or other external
structures and complicated insides? 11 4
4. Do the cells contain internal bodies
(usually green or golden brown) called
chloroplasts? (Sometimes these cells
are contained inside outer covers which
may hide them completely or partially.
Sometimes they have red "eyespots" or
long slender hairs called flagellae.) 6 5
5. Are the cells without any, or at least
with very little visible, internal
structure? Generally, these cells are
bluish-green in color (especially a mass
of them together) and are very minute. If
so, they are Blue-Green Algae. See Plates
I, II. - 3
A-69
-------
Appendix I
Yes No
6. Do these tiny plants consist of single
cells or groups of cells which move about
by means of one or more long slender hair-
like "flagellae?" Red eyespots usually can
be seen.* If so, they are Flagellates.
See Plates V, VI.
*0ne kind is large and filled with rust-
red granules. -- 7
7. Are they golden brown in color, with a
tendency to sharply angular edges? They
may be cylindrical, thread-like or
boat-shaped. Some may move in a hesitant
manner. If so, these are Diatoms. See
Plates VII, VIII. - 8
8. These plants are all green. Do they
consist of single cells or small
clumps of 2 to 4 cells, but not long
strings or filaments? (No flagellae
or movement should be observed. If it
is, return to 6.) These are Coccoid Green
Algae. See Plate III. -- 9
9. The following plants (9 and 10) are all
filamentous or thread-like, and consist
of single cells; cylindrical, barrel-
shaped, or roundish, attached end-to-end.
Is green pigment (chlorophyll) contained
in various shaped bodies within the cells
(chloroplasts)? If the mass of
filaments appears green or yellow-green to
the naked eye, they are Filamentous Green
Algae. See Plate IV. — 10
10. Are the cells of the filament apparently
without internal structure (although
confused "pseudovacuoles" may sometimes
be seen)? Larger oblong cells with heavy
walls (heterocysts) or apparently empty
cells (akinetes) may occur. Some types
with smooth surfaces may move slowly but
visibly. Mass of filaments appears some
shade of bluish-green or red to the naked
A-70
-------
Appendix I
Yes No
eye, occasionally very dark. If as above,
these are Filamentous Blue-Green Algae.
See Plates I, II. — 4
11. Is the body composed of a single cell or
unit? It may be enclosed in a shell or
sheath. It may bear cilia over all or
part of the body; flagellated forms may
also be encountered: Protozoa. See
Plates IXa and IXb. -- 12
12. Is the noncolored body of the organism
composed of many cells, with well organized
internal structures? There are often one
or two crowns of tiny hairs or cilia
(which can rarely be seen individually) at
one end. One common type crawls like an
"inchworm," or "accordion"; others have
flattened shells, spines, or other features,
but there are no true legs. If as above,
this is a Rotifer. See Plate X. — 13
13. Does the specimen have jointed appendages
(joints in the body can also usually be
seen), usually with characteristic hairs
or setae (may not be extended unless
animal is active)? 14 3
14. Is the body completely enclosed in two
minute clam-like shells? Jointed legs
may be extended from between shells for
swimming. These are microcrustaceans,
Ostracods. See Plate XII. -- 15
15. Does the elongated, segmented, trans-
parent body (which may range up to
approximately 1/2 inch in length) have a
head with two eyes? If so, this is a
phantom midge larva, an insect:
Chaoborus. See Plate XII. -- 16
16. Does the organism have a single eye
and two shell-like projections that come
down on either side of the legs? Eggs
may be present in a large pouch inside
the upper back part of the shell.
A-71
-------
Appendix I
Yes No
This is another microcrustacean, a water
flea: Cladocera. See Plate XI. -- 17
17. Is there a single eye, the body
relatively cylindrical, no side shells,
usually tapering toward the rear?
There may be segments (or joint)
in the body. Two large front "legs"
(actually antennae) are used for
locomotion. This is another micro-
crustacean, a Copepod. See Plate XI. -- 18
18. Is the body similar to the above, but
roundish or pear-shaped, smaller, and
with no segmentation or joints in the
body? There are three pairs of
relatively large legs. This is the
larva of a copepod called a Nauplius.
See Plate XI. — 11
A-72
-------
Appendix I
Blue-Green Algae Myxophyceae
Oscillatoria spp., filaments (trichomes) range from .6 to over
60 11 in diameter. Ubiquitous, pollution tolerant.
Lyngbya spp., similar to Oscillatoria but has a sheath. A, Lyngbya
contorta, reported to be generally intolerant of pollution; B, L.
birgei.
Plate I a.
A-73
-------
Appendix I
Blue-Green Algae Myxophyceae
Aphanizomenon flos-aguae A, colony; B, filament
Anabaena flos-aquae A, akinetej B, heterooyst
Plate I b.
A-74
-------
Appendix I
borne Blue-Green Algae
I. Nonfilamentous (coccoid) Blue-Green Algae:
Anacystis (Chroococcus) (X600)
Agnienellum
(MerismcpediuraJ (X600)
Coccochloris (Gloeocapsa)( X600i Microcystis (X600) Polycystis
Plate II a.
A-75
-------
Appendix I
Some Blue-Green Algae
II. Filamentous blue-green algae:
Trichomes of Spirulina. (X600)
Trichomes of Arthrospira (X600)
Phormidium (with she-
athl
Oscillatoria (without
sheath) (X825)
Anabaena (X82£)
True branching
Hapalosiphon (X375)
False branching
Tolypothrix (X375')
Plate II b.
A-76
-------
Appendix I
Nonmotile Green Algae: Coccoid
(Chlorophyceae)
Pediastrum
Species of the Genus Scenedesimis
S. caudatus
S. abundans
S. dimorphus
Plate III a.
A-77
-------
Appendix I
Nonmotile Green Algae: Coccoid
(Chlorophyceae)
Desmids
go
Closterium
Cosmarium
Staurastrum
Plate III b.
A-78
-------
Appendioc I
Nonmotile Green Algae
(Chlorophyceae)
A-79
-------
Appendix I
Flagellated Algae
Goniaulax
Ceratium
Eudorina
Plate V a.
A-80
-------
Appendix I
Flagellated Algae
Trachelomonas
Chlamydonionas
Mallomonas
Dinobryon
A, form of colony; B, cell in
lorica.
Plate V b.
A-81
-------
Appendix I
General Morphology Of Algal Flagellates
Greens
Phacus Trachelomonas
Euglena gracilis Lepocinclis
Greens
Carteria V Gonium Sociale
Chlamydomonaj^ Chlorogonium
Colony of Volvox
Gonium Pectorale Colonj Pandorina Colony
Plate VI a.
A-82
-------
Appendix I
General Morphology of Algal Flagellates
Yellows
Chromulina
Synura Dinobryon Lorica
Browns
Peridinium Gyrnnodinuim
Plate VI b.
A-83
-------
Appendix I
Diatoms - Bacillariophyceae
Valve views
Girdle views,
Stylized to show
basic diatom
structure.
A discoid or central
diatom such as
Stephanodis ous
Plate VII a.
A pennate or navicular
datom such as
Synedra
A-84
-------
Appendix I
Diatoms - Bacillariophyceae
A colony of Asterionella
(girdle~views)
A co?.ony of Fragillaria
(girdle^views)
A
.nil 1 II IU
Gomphonoma
A, valve view; B, girdle view.
IL Tl
Diagram showing progressive diminution in the size of certain
frustules through successive cell generations of a diatom.
Plate VII b.
A-85
-------
Appendix I
flVlft '. * r.*~**r*?*-* 1 .<:"•** ----» v» •
RHIZCSOLENIA
ASJERIONE1LLA
CYtLOTELQ-3
-------
Appendix I
Phylum Protozoa
Class Mastigophora, the flagellates
Bodo
Pollution tolerant
19 i
Peranema
(moderately pollution tolerant)
Anthophysis
Pollution tolerant
Naegleria
(Pollution tolerant;
Colony of Poteriodendron
Pollution tolerant, 35 >
Plate IX a.
A-87
-------
Appendix I
Phylum Protozoa
Class Sarcodina, the amoebas
Some forms often found as plankton:
Ameba
Top
Side
Arcella
(Pollution tolerant)
Plate IX b.
Actinosphaerium
(Pollution tolerant)
up to 300 i
A-88
-------
AppencLLx I
Phylum Protozoa
Class Ciliophora, the ciliates
Euplotgjs
Pollution tolerant, 90
Fodophrya, a suctorian
ciliate. Pollution
tolerant. 20-^0 i
Central View
Side View
Golpoda
Pollution tolerant
20-120ji
Holophrya, reported
to be intolerant of
pollution, 35 V-
Plate IX c.
Epistylis, pollut-
ion tolerant. Col-
onies often macro-
scopic.
A-89
-------
Appendix I
Phylum Protozoa
Class Ciliqphora, the ciliates
Some forms often found as plankton:
Yorticella
(Pollution tolerant)
Head 75 - 100 i
Codonella
60 - 70^1
Tintinnidium
100 - 2uO
Plate IX d.
A-90
-------
Appendix I
Plankton!c Rotifers
Various Forms of Keratella cochlearis
Philodino Rotaria type
Plate X a.
A-91
-------
Appendix I
Plankton!c Rotifers
Various Forms of Keratella Cochlearis
Synchaeta
Brachionus
Polygarthra
Plate X b.
A-92
-------
Appendix I
Class Crustacea
Fairy Shrimp;
Eubranchipus, Order
Phyllcpoda
20-25 mm,
Crayfish, or crawdad;
Cambarus, Order Deoapoda
5-15 cm.
Aquatic Sow Bugj Asellus,
Order Isopoda
10-20 miru
Water Flea
Daphnia
Order
Cladocera
Plate XI a.
A-93
-------
Appendix I
Class Crustacea
Scudj Hyalella,
Order Amphipoda
10-15 mm
Fish Louse^ Argulus,
a parasitic Copepod
5-6 rron
Copepod; Cyclops, Order Copopoda
2-3 ram
Plate XI b.
A-94
-------
Appendix I
Left: Shell closed
Ostracod
Right: Appendages extended
A Nauplius larva of a Copepod
Chaoborus midge larva
Plate XII
A-95
-------
Appendix I
3. Biological Field Methods
Biological field activities usually consist of two major
activities: the collection of specimens and the recording
of careful observations. Compact kits of field collecting
equipment and materials greatly increase efficiency, especially
if the collection site is remote from transportation. All
collecting containers should be identified with location,
station number, sample number, and the date. Much time may be
saved by using data sheets or cards with uniform arrangement for
entering the data. The same data sheet may include laboratory
or field analysis. Sample data sheets are included at the end
of this section. Field notes should be taken in pencil to
preserve them in case they get wet.
Observations of the general biological and physical
characteristics of the sampling site should be recorded before
any sampling is done. Underwater swimming or the use of scuba
may be valuable in certain locations for direct observation
and collecting. Underwater and aerial photography may be
useful.
Because of the diverse nature of aquatic organisms, different
methods of collection are used for the various kinds. Aquatic
mammals and birds usually require other approaches and are not
included. Collection methods for oceanic, estuarine, or fresh-
water situations are similar. Marine organisms range to larger
sizes than those of freshwater. Because of the corrosive
nature of sea water, special care should be taken in the design
and maintenance of collecting equipment. Site selection and
collection schedules for marine sampling are influenced by
such factors as tidal currents and salinity distribution
rather than river currents, riffles, and pools. Lake collection
usually shows less predictable flow patterns. Before going into
the field, the investigator should decide on the size range of
the organisms to be collected (microscopic, macroscopic) and the
kind of organisms (invertebrates, vertebrates, vascular or
nonvascular plants) which he will seek.
The following sections explain the collection methods for four
groups of aquatic life:
a. Benthos
These are bottom dwelling organisms. They may be attached,
crawling, or burrowing forms. Some of the collecting
devices are shown in Plates I and II. In most instances,
home-made equipment can be substituted for the standard
research type. Hand picking of benthic organisms from
rocks, sticks, etc., that have been picked out of the
water is a fast and much used method for quickly
A-96
-------
Appendix I
determining what is present and what might be expected
in additional samples.
Patches of seaweed and eelgrass and shallow weedy margins
are most often studied on a qualitative basis only. The
apron net is used for collections in weed beds or in other
heavy vegetation. It is simply a pointed wire sieve on a
long handle with coarse screening on the top to keep out
leaves and sticks. Poking it into and then withdrawing it
from the weed masses is the method of operation.
Masses of weeds may be pulled out on the bank (with rakes,
grappling hooks, etc.). The benthic organisms can then be
observed as they crawl out.
Quantitative estimates of both plants and animals can be
made by using a "stove pipe" sampler. This is a hollow tube
which is forced down through the weed mass in shallow water
and embedded in the bottom. The contents can then be removed
and placed into a series of sieves for sorting.
A frame of known dimensions can be placed on the bottom, and
the material within is then cropped out. This is especially
good for larger plants and for large bivalves. It is also
useful on sand and mud flats.
Handle-operated samplers, such as the Jackson, are
effective for sampling a variety of bottoms down to the
depth of the handles. Such samples are then washed through
graded screens to retrieve the organisms.
The Ekman Dredge is a device which is used to collect bottom
samples.It should be used in bodies of water which have
muddy or sandy bottoms. It will not work well on gravel
or rocky bottoms.
The dredge is lowered into the water until it comes to rest
on the bottom. In shallow water, you can place it on the
bottom; for deeper water, you lower it on a line or a stick.
Next the spring mechamism is tripped. This is done by hand
in shallow water and by using the messenger (a device which
comes with the dredge) in deep water. After the spring
is released, the jaws snap shut and enclose the sample.
Finally, bring the dredge to the surface and empty the
sample into a plastic bag. Refrigerate or cool the bag if
the sample will not be studied within an hour. (Benthic
organisms decompose rapidly in warm weather.)
Dump the sample out onto the top of a series of graduated
mesh, brass screens. The screen with the largest mesh (size
of openings) should be on the top and that with the smallest
A-97
-------
Appendix I
on the bottom. Mesh sizes in between should be arranged in
order of decreasing mesh size. Flood the sample with water.
(Stirring the sample may be helpful.) This procedure will
effect two sortings according to size, "soil" partic'les and
macroinvertebrates. Use forceps to collect the macro-
invertebrates. Count, identify, mass, and preserve them.
You may want to compute the density and the biomass of the
macroinvertebrates and relate these to other parameters of
your study. You may want to study the relationship between
"soil" particle size and type and the macroinvertebrates
present. An Ekman Dredge can be ordered from Wildlife
Supply Company, Saganaw, Michigan and other companies. The
cost is about $60.
The Petersen type, which grabs without weights, will take
satisfactory samples in firm muds but tends to bury itself
in very soft bottoms. It is seldom used in shallow water
except as noted below.
The riffle (rift) is one of the most satisfactory habitats
for comparing stream conditions at different locations. The
hand screen is the simplest and easiest device to use, but
the resulting collections are qualitative only. The screen
is firmly placed in the stream bed. The upstream bottom is
thoroughly disturbed with the feet. The current carries the
organisms to the screen. The screen is then lifted, and the
contents are dumped into a sorting tray or collecting jar.
The Surber Square Foot Sampler is one of the best
quantitative collection devices for rifts. It is firmly
planted on the bottom. The stones and other material within
the square frame are carefully rubbed by hand to dislodge
all benthic organisms. The current carries them into the
net. A stiff vegetable brush is often useful, especially if
the bottom materials are covered with moss. When bottom
materials are picked up which are free from macroinvertebrates,
the sampling is finished. Before removing the sampler from
the water, the bottom should be "fanned" with the hand to
kick up any macroinvertebrates which may have fallen straight
down rather than being carried into the net. The organisms
are then removed from the net and placed in a plastic bag or
a collection bottle. To insure a representative sampling,
3 to 5 square foot samples should be taken at each location.
A Petersen type grab may be used in deep swift riffles. It
is placed on the bottom and worked into place with the feet
or with poles. After being closed, it is lifted by pulling
on the rope in the usual manner.
A-98
-------
Appendix I
A strong medium-weight dipnet is the closest thing to a
universal collecting tool. Collections are made by sweeping
through weeds, over the bottom, or in open water. The
handle should be from 4 to 6 feet long and about the weight
of a garden rake handle. The rim should be made of steel or
brass. The size of the rim stock will depend on the size of
the rim; it should be strong but not cumbersome. The netting
should be the strongest available preferably with about a
1/16 inch mesh. Nets which are too fine plug up easily and
cannot be moved quickly through the water. The net must be
protected around the rim. This can be done by sewing canvas,
leather, or pieces of old innertube around the rim.
When sampling from vessels, a crane or winch is often used.
The general ideas described for shallow water often apply
also to deeper waters. The Petersen type grab is probably
the best all-around sampler for the greatest variety of
bottoms at all depths, from shoreline down to over 10,000
meters. If hauled by hand, the grab should be fitted with 5/8
or 3/4 inch diameter rope in order to provide an adequate
hand grip. It is best handled by means of wire ropes and a
winch.
Drag dredges or scrapes (Plate I) are often used in marine
waters. They have not been used to any great extent in
freshwater studies.
Since most biological communities are not evenly distri-
buted, one should routinely take at least two, and
preferably more, samples from any one site.
Artificial substrates (growing surfaces) are also used in
studying benthic organisms. When an artificial habitat is
exposed in a given site for 2 to 3 weeks, it tends to become
populated by all available species partial to that type of
habit. These devices can then be collected and taken to the
laboratory for evaluation. They consist of such items as
cement plates and panels, wood (especially for burrowing
forms), glass, microscope slides (Catherwood diatometer),
the Hester-Dendy Sampler, baskets holding natural bottom
material, ropes suspended in the water, and sticks thrust
into the bottom. The Hester-Dendy Sampler is easily made
and lends itself well to student use; therefore, it is
described in detail below.
The Hester-Dendy Sampler is used to collect benthic (bottom-
dwelling) macroinvertebrates. It can also be used to collect
attached algae and some types of diatoms. This sampler has
.0929 square meter (1 square foot) of exposed surface.
A-99
-------
Appendix I
Ekman
Bottom Grabs
-J^ P
Jackson
Plate I
A-100
-------
Appendix I
Limnological Equipment
Hand Screen
Surber Sampler
Specimen or
reagent bottles
Apron net
Sorting pan
Slurrey bucket
Pail
Plate II
A-101
-------
Appendix I
Deep Water Equipment
Biological dredge
Plate III
A-102
-------
Appendix I
The materials needed are 1/4 inch and 3/16 inch hardboard,
threaded 1/4 inch steel rod, and nuts. Cut the 1/4 inch
hardboard into 1 inch squares and the 3/16 inch hardboard
into 3 inch squares. Drill 3/8 inch holes in the centers
of all the squares.
Assemble the sampler by sliding one of the 3 inch squares
on the steel rod, and add one of the 1 inch squares. Con-
tinue this procedure until you have added nine more of the
3 inch squares, and eight more of the 1 inch squares. Now
add a nut to each end of the steel rod and tighten. A nylon
cord may be used in place of the steel rod.
Tighten the nuts until no space remains between the 1 inch
and the 3 inch squares and until the squares will stay in
place. (An extra 3 inch square may be added without a
spacer if you wish to determine which microorganisms might
attach themselves to the nonexposed surfaces.)
For any investigation, either place all samplers in flowing
water or place all samplers in still waters. (Still waters
and flowing waters usually have different benthic populations.)
The samplers may be placed on the bottom if it is composed of
sand, gravel, or rock. If the bottom is made of mud, suspend
the sampler just off the bottom. (If this is not done, the
sampler may become covered with mud and the collected
benthic sample will not be representative.)
If the samplers are placed in highly populated or well used
areas, they should be hidden so that they will not be
disturbed. In most locations, the samplers should be tied to
an overhanging branch, to a tree root, or to the bank. Heavy
(30 pound test or higher) monofilament fishing line will be
less visible than most kinds of string. In very swift water
or in locations where attachment is difficult, the sampler may
be attached to a metal rod which has been driven into the
bottom of the stream or lake.
To show the effects of an effluent on the benthic macro-
invertebrates, place samplers upstream and downstream from
the point at which the effluent enters. Samplers should be
placed on both sides of the stream; for the larger streams
and rivers, they should also be placed in the middle.
After the samplers have been in the water for 2 weeks or
more, they should be collected. Immediately after removing
each sampler from the water, place it in a plastic bag and
add some surface water from which the sampler came. This
will prevent the loss and drying out of the organisms.
A-103
-------
Appendix I
If more than an hour will elapse before you begin to
identify the organisms, the plastic bag containing the
sampler should be cooled with ice or should be refrigerated.
This will prevent the organisms from decomposing, which can
happen very rapidly, especially in hot weather.
Open the plastic bag over a white porcelain tray and remove
the water and the sampler. Disassemble the sampler and
scrape off any macroinvertebrates which are still attached.
(A laboratory spatula works well for the scraping.) If
large numbers of organisms are present, remove and collect
them from one 3 inch square at a time. This will make them
easier to count and to identify. (This writer has collected
one sampler which had more than 1,300 organisms on it.)
The organisms may be preserved in alcohol or formalin for
future reference.
The results should be used to compute diversity. The biomass,
mass of the life in a specified unit of the environment
(1 square foot, in this study), can also be computed. This
would give an indication of the productivity of the water.
Benthic collections often consist of large amounts of debris.
Various procedures may be followed to separate the organisms
from the debris. This separation may be done by hand
picking, which is best done on a white enameled tray using
light touch limnological forceps. Screening is one of the
most practical means of separation. The sample may be dumped
onto the screens, and then separated by pouring water over it
to wash away the mud and debris. Another method is to place
the sample in a bucket or tub and then add water. The mix-
ture is swirled vigorously, and the supernatant is poured
through the screen. The residue should be examined for
heavier forms which did not float to the top. A variation
of this method is to pour a salt or sugar solution Into the
bucket. The mixture is stirred well, and the supernatant is
poured through the screen (save it for reuse). The denser-
than-water solution effects the separation of organisms from
the debris. A solution of 2-1/2 pounds of table sugar per
gallon of water is considered to be optimum for most samples.
Preservation of samples may be achieved by placing them in
80% ethyl alcohol in the field. For prolonged storage, they
should be placed in a fresh solution of 70% ethanol. Formalin
is also effective in 3% to 10% solutions of the commercial
form. Odor and shrinkage problems exist with this pre-
servative. Neutralized formalin eliminates some of the
undesirable effects. For short-term preservation,
refrigeration and icing are adequate.
A-104
-------
Appendix I
b. Periphyton or Aufwuchs
Periphyton is the collection of organisms attached or
clinging to stems and leaves of rooted plants or other
surfaces projecting above the bottom of an aquatic system.
It consists of algae, fungi, small animals, and protists.
Periphyton is sometimes referred to as the slime forming
organisms.
One qualitative method of collection is to scrape the
periphyton from surrounding surfaces, and to place the
scrapings in a 4% formaldehyde solution. This can be
quantified somewhat by scraping all surface material from
a measured area. A more effective quantitative procedure
would be to collect the periphyton on an artificial sub-
strate such as glass microscope slides suspended in the
water as described above.
By using a microscope and the appropriate identification
keys, one can identify the periphyton in the sample.
c. Plankton
Plankton (plancton) is defined as all the microscopic
plants, animals, and protists normally swimming or suspended
in open water.
A comprehensive plankton sampling program would involve
sampling at weekly or more frequent intervals. A year-long
study of this type would provide valuable data which could
be used to predict conditions in following years.
Phytoplankton (algae) can be collected at the surface in
half-liter bottles. For deeper samples, a Kemmerer, Nansen,
or other specialized collector may be used. A plankton net
is also useful. Sizes number 20 or number 25 are commonly
used for collecting phytoplankton. Nets concentrate the
organisms in the process of collecting-, however, the smaller
forms will be lost through any net.
Zooplankton (animals and protists) have the ability to swim
away from a collection bottle; so they are best captured
with nets which are towed at moderately fast speeds. Number
12 nets (operative size 0.119 mm, 125 meshes per inch) or
smaller numbered net sizes are commonly used. The mesh size
of the net determines the size of the plankton to be
collected.
Both shallow and deep samples are suggested. Shallow
samples are taken at a depth of 6 inches to 1 foot. Surface
film is also often significant. Deep samples should be
A-105
-------
Appendix I
taken at as many locations between the surface and the
bottom as the study demands. The most complete study would
sample the entire water column and would record the kinds of
plankton found at each level.
Estuarine plankton should be sampled at different stages of
the tide. Since plankton is affected by the forces of winds
and currents, a tow is often best made at right angles to the
direction of wind or current.
Zooplankton tend to collect near the bottom in daylight
and to distribute more evenly at night. One method commonly
used to get a representative sample is to take an oblique
tow from the bottom to the top of the water column.
Field conditions greatly affect plankton, and they should
be carefully noted on the field data card.
Unless the samples will be analyzed within an hour after
collection, they should be stabilized in the field.
Refrigeration or icing is very helpful, but do not put the
ice j[n_ the sample. A 5% formalin solution is often used, but
it shrinks animals and makes all forms brittle. Lugol's
solution is a good preservative. Ultra-violet sterilization
is sometimes used to retard the decomposition of plankton.
A good methiolate preservative has been developed by the
FWQA; it has been described by Weber (1968).
d. Nekton
The larger, free swimming animals such as fish, shrimp,
and eels are called nekton. To insure a representative
sample, they must be collected from the obscure and unlikely
areas as well as the obvious. A check should be made with the
local authorities before the sampling is done because many
of the standard techniques that are used are not legal for
the layman. Professionally trained workers are very
important in this area of investigation than perhaps in any
other area.
The various devices include haul seines, gillnets, trap nets,
traps, trawls, and electrofishing apparatus.
Personal observations by competent personnel and informal
inquiries with local residents often yield valuable infor-
mation. The organized creel census yields data on what
kind and how many fish are being caught.
Fish and other nekton are sometimes tagged or branded to
trace their movements during migration and at other times.
A-106
-------
Appendix I
Miniature radio transmitters can be fed to or attached to
them and the nekton can be tracked over considerable
distances. Physiological information can be obtained in
this manner, also. This is known as telemetry.
e. Sample Data Sheets
Examples of data sheets appear on the following pages.
These can be reproduced easily; if desired, they can be
punched or stapled into notebooks.
A-107
-------
Appendix I
LAKE, IMPOUNDMENT, OR ESTUARY
SURVEY
Location and General Characteristics
Reporter:
1. Name:
County: Township:
Nearest Town:
Map agency: Name No.
2. Observed nearby land use:
3. Maximum drawdown or tidal range
4. Depth: Average Maximum _
5. Area: Shoreline Length
6. Shoreline development*:
7. Watershed size:
8. Nature and extent of erosion observed:
9. Possible pollution sources:
10. Study Station No. : Description:
- S
2 *J a.'W
S = Shoreline length
a = area L-l
A-108
-------
Appendix I
GENERAL MAP
Showing station locations
A-109
-------
Appendix I
LAKE, IMPOUNDMENT, OR ESTUARY
SURVEY
Station:
Present Conditions
Date:
1. Weather
a. Cloud cover:
b. Wind directionT
c. Air Temp:
d. Other:
2. Waves
a. Height:
b. Other:
Length:
Time:
Velocity:
Precip:
Fetch:
3. Ice, thickness:
Condition:
4. Floating materials:
5. Water Color:
6. Nature and origin of color:
7. Odor, if distinctive:
8. Secchi disc:
or
Turbidity rod:
9. Notes:
L-2
A-no
-------
Appendix I
Station:
LAKE, IMPOUNDMENT, OR ESTUARY
SURVEY
Temperature Profile
Date: Time:
Temperature
8 12 16 20 24 28 32 36 40
ri
o
CO
T3
•o
-------
Appendix I
Station:
*d o
LAKE, IMPOUNDMENT, OR ESTUARY
SURVEY
Physical and Chemical
Date: Time:
o>
o crp
D
o>
T3
0)
>-5
d
0)
<-h
0)
P
t-l-
0)
0)
ft-
o
a
(D
a
L-3b
A-112
-------
Appendix I
LAKE, IMPOUNDMENT, OR ESTUARY
SURVEY
Station:
•o o 3 S
• • • iJ
Physical and Chemical
Date: Time:
_, ^ <_i. M. y oq .^ re a. o a"
3*
re
M
O
re
r-t-
0)
1
3
>-"•
3
P3
rt-
M-
o
5
CO
fD
re
c*
3-
o
a
05
re
a
L- .
A-113
-------
Appendix I
LAKE, IMPOUNDMENT, OR ESTUARY
SURVEY
Biological Data - Plankton
Station: Date: Time:
Survey Counts
Sample No.: Type: Depth:
Procedure:
Results:
Sample No. : Type: Depth:
Procedure:
Results:
L-4a
A-114
-------
Appendix I
LAKE, IMPOUNDMENT, OR ESTUARY
SURVEY
Biological Data - Plankton
Station: Date: Time:
Qualitative, Differential, or Proportional Counts
L-4b
A-115
-------
Appendix I
LAKE, IMPOUNDMENT, OR ESTUARY
SURVEY
Station:
Biological Data - Fish
Date: Time:
How collected:
Sample No.
E Weight
No. of fish
Other:
Dominant Kinds
Sample No.
Other:
E Weight
Dominant Kinds
No. of fish
L-5
A-116
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Appendix I
AQUATIC VEGETATION MAP
A-117
-------
Appendix I
LAKE, IMPOUNDMENT, OR ESTUARY
SURVEY
Station:
Biological Data - Bottom Forms
(Benthos and Periphyton)
Date:
1. Aquatic Vegetation
Kinds of Plants
Extent of Coverage
How collected:
Nature of bottom:
2. Periphyton
Kinds
Extent of Coverage
Description:
How collected:
Nature of bottom:
3. Attached Algae
Kinds
Extent of Coverage
How collected:
Nature of bottom:
L-6
A-118
-------
Appendix I
BOTTOM TYPES MAP
A-119
-------
Appendix I
LAKE, IMPOUNDMENT, OR ESTUARY
SURVEY
Biological Data - Bottom Forms
(Benthos and Periphyton)
Station:
Date:
Time:
Sample No. Unit
No. Spp.
No. Ind.
1. Insects
Kinds
No. or Rel.
Abundance*
2. Other Invertebrates
Kinds
No. or Rel.
Abundance*
How collected:
Nature of bottom:
* + = present
c = common
a = abundant
d = dominating
L-7
A-120
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Appendix I
LAKE, IMPOUNDMENT OR ESTUARY
SURVEY
Station:
Bacteriological Data
Date:
Time:
How and where collected:
Time collected:
Collected by:
Tests started
5. m.
Temp:
a. m.
-p. m.
--,
By:
Tests requested:
(check)
Coliform: _
Fecal
Coliform:
Fecal
Streptococcus:
Remarks:
Test results: Coliforms:
Fecal
Coliforms:
Fecal
Streptococci:
Method: Membrane Filter ( )
100 ml.
100 ml/
100 ml.
L-8
A-121
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Appendix I
NOTES
A-122
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Appendix I
4. Biological Laboratory Methods
Living specimens are the most desirable for laboratory analysis.
Unfortunately, the investigator must often work with preserved
specimens. When conducting a comprehensive investigation, a
biologist usually collects samples faster than he can analyze
them.
After the laboratory data have been recorded, they should be
interpreted with reference to the field notes which were taken
at the time of sample collection.
a. Benthos
Although some fish (i.e., flounders) are listed as benthic
organisms, the usual bottom sample is analyzed for the
macroinvertebrates which it contains. After the foreign
material has been removed from the sample (see Section 3),
the investigator counts, identifies, and determines the mass
of the organisms.
A technique commonly used is to pour the sample into a white
porcelain tray which has some tap water in it. Laboratory
forceps are usually used to remove the organisms. If the
sample contains large numbers of organisms, the investigator
will find a mechanical hand counter very useful. The
organisms should be placed on blotting paper for 1 minute
before the mass determination is made.
If the investigator does not wish to "key out" the organisms,
he may simply sort and group on the basis of like appearance.
He might sketch and/or describe the different kinds and report
something like "75 individuals of taxon (or type) 1."
b. Periphyton or Aufwuchs
Direct analysis of the growths attached to the substrate can
be carried out but must be restricted to the larger organisms.
This is due to the difficulty of keeping the material in an
acceptable condition under the short working distances of the
objective lenses of compound microscopes and due to the fact
that transmitted light is not adequate when the colonial
growths are thick or the substrate is opaque.
More often the periphyton is scraped from the substrate and
then processed. An aliquot part of the sample may be
counted using methods frequently employed in plankton analysis.
The number of organisms per unit of volume can then be
determined.
A-123
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Appendix I
The total dry weight of the scrapings and the ash-free
dry weight (which eliminates inorganic sediments) can be
determined and compared. A packed biomass and volume could
be determined by centrifugation of the scrapings. Nutrient
analyses serve as indices of the biomass by measuring the
quantity of nutrient incorporated. Total organic carbon,
carbon equivalents (COD), and organic nitrogen
determination would be helpful. Phosphorus has limitations
because cells can store excess above immediate needs.
Chlorophyll and other bio-pigment extractions might be
carried out to determine the amount of these which are
present. Other investigations might measure carbcn-14 up-
take, oxygen production, or respiratory oxygen demand.
Qualitative studies would produce such results as the kinds
found, ratios for number of individuals per kind found, and
a frequency distribution of varieties found.
Quantitative investigations could yield amounts peir unit
area, milligrams per square centimeter. Rate studies could
determine such things as milligrams per day of biomass
accumulation or milligrams of oxygen produced per milligram
of growth per hour.
c. Plankton
Microscopic examination is most frequently done in the
laboratory to determine the number and kinds of organisms
present. Optical equipment need not be elaborate for
qualitative studies. If more precision is required, such
items as a Whipple counting eyepiece, a mechanical stage,
and a stage micrometer may be used.
Precision-made counting chambers such as Sedgewick-Rafter
counting cells, Palmer-Maloney counting cells, or haemo-
cytometers are required for quantitative work with liquid
mounts. Qualitative "counts" are lists of the kinds of
organisms found and the numbers of each per unit of volume
or area.
The organisms are observed and, by means of a suitable
series of multiplier factors, projected to a number or mass
per unit volume. Counting of an unconcentrated sample
eliminates manipulation. If the density of organisms is
low, more area can be examined or the sample can be con-
centrated. The concentration of the sample provides more
organisms for observation, but this introduces additional
errors and takes more time.
Several methods of counting are in general use. The
numerical or clump count is regarded as the simplest. The
A-124
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Appendix I
area! standard unit method (See Standard Methods) provides
more information. The cubic standard unit method is a logical
extension of the area! method, but it has not achieved wide
acceptance due to its difficulty.
The field count is done by counting and tallying all
individuals of each type present in a field of view. A good
way to do this is to list the most common types separately,
record their counts, and enumerate the other forms present.
This is done for five or ten randomly chosen fields. Finally,
the results are tallied and the percentage of each type is
computed.
The Five Hundred Count is done by moving the slide at random
and counting and tallying all the types until a total of 500
cells or clumps have been counted. Then the investigator
should tally the results and compute the percentage of each
type as before.
Sometimes, measurements are made by means other than micro-
scopic counts. Settled volume of killed plankton may be
measured in an Imhoff cone or a graduated cylinder after a
standard period of time. This will evaluate only larger
forms. A gravimetric method involves drying at 60° C for
24 hours followed by ashing at 600° C for 30 minutes. This
is particularly useful for chemical and radiological
analyses.
Chlorophyll can be extracted by filtering, drying for 24 hours,
and extraction with methyl alcohol. Evaluation can be made
by using a colorimeter or by using chromatographic methods.
A membrane filter may be used. The filter can be cleared with
immersion oil and organisms can be observed directly after 24
hours, or the collected material can be washed off and
observed immediately.
d. Nekton
Population studies are often done with the larger animals.
The individuals should be checked for general condition and
for the presence of parasites.
As mentioned before, the collection of fish is best done by
professionals.
e. Bioassays and Biomonitoring
The bioassay technique may be used on any appropriate
organisms from protozoa to fish. In the lab, two types of
apparatus, the static jar and the continuous flow, may be
employed to provide the various dilutions of the toxicant
used.
A-125
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Appendix I
Static jar tests are containers with a known concentration
of substance and organisms. These tests are seldom run for
more than 1 week and are read only in terms of percent;
survival or kill. This is usually termed acute toxicity.
The continuous flow apparatus may also be used to measiure
acute toxicity. This setup allows a solution to flow into
the containers at certain time intervals. Continuous flow
apparatus is virtually essential for long-term tests at sub-
lethal concentrations. Parameters other than lethal
thresholds can then be measured, such as the effect on
growth rate and breeding success of a species of fish.
Biomonitoring permits continuous surveillance over the
toxicity of an effluent. This technique involves the
placing of living organisms in test waters. A normal length
of time to run the test in water pollution studies is
96 hours. For the results to be significant, a 50% or
greater kill must occur during the 96 hours.
In the laboratory, place ten middle-sized Daphnia in each
of several 4 ounce bottles. Daphnia magna, commonly called
water fleas, are small crustaceans. If they cannot be
obtained locally, they can be ordered live from a biological
supply house. Middle-sized ones are used so that natural
mortality due to age, will not lead to incorrect results.
Cover each bottle with nylon cloth having at least 80
threads to the inch. If this kind of cloth is not available,
two thicknesses of a piece of nylon stocking will probably be
acceptable. The cloth must detain the Daphnia and permit the
dissolved substances to diffuse into the bottle. Use rubber
bands to fasten the cloth over the opening of the bottle.
Place five of these bottles in a 6 inch square wire
container (which can be made from 1/4 inch hardware cloth)
and place it in the stream. After 96 hours, count arid record
the number of living adults and offspring.
Small (2 to 3 inch) bluegills (Lepomis macrochirus) and/or
largemouth basses (Micropterus salmoides) can" also'be used.
These should be fed and acclimated to~Taboratory conditions
for 2 to 4 days before being placed in the stream.
The fishes can be transported to the bioassay stations in 10-
gallon milk containers lined with large plastic bags. If
aeration is necessary, use an aquarium pump which can be run
off an inverter. The inverter changes 12 volts into 110 volts,
At each station place, 10 bluegills or 10 largemouth basses
in a Gee's galvanized, quarter-inch-square wire minnow trap
(manufactured by Cuba Specialty Manufacturing Company,
A-126
-------
Appendix I
Houghton, New York). Close the hole in each end of the
minnow trap with a cork or a rubber stopper. This will
prevent predators, such as eels, from getting into the trap.
Slowly acclimate the fish to the temperature of the stream.
This can be done by putting the fish into a plastic bag
(with water from the milk can) and then placing the plastic
bag into the stream or else by adding stream water--small
amounts at a time—to the plastic bag.
After adding the fish to the trap, place it in 1 or 2 feet
of water of very low velocity. If slow-moving water cannot
be found, place it on the downstream side of a large rock or
other obstruction.
Transfer the living fish from the trap to a plastic bag
(after 96 hours). Note the conditions of the surviving fish.
Look for vitality or a lack of it. Observe the fins. Some
chemical effluents cause them to deteriorate. Record the
number of dead fish and the number and conditions of each of
the surviving fish.
If these tests are done in highly populated or well used
areas you must carefully hide the bottles and traps. If
this is not done, they will be disturbed or taken.
Be sure that you do not subject the Daphnia or the fish to
temperature shock when placing them in the stream. Daphnia
can stand a rapid temperature change of only one to 2o F and
the fish can stand a rapid change of only 2° F to 4° F.
f. Diversity Indices
The statistical analysis of some kinds of biological data
may be done by computing the diversity index. Generally
speaking, the greater the diversity, the healthier the
biotic community. There is presently no one method of
computing this index, which is universally accepted by the
professionals. The methods range from simple ones to others
that are best calculated by computers.
The sequential diversity index is explained below. An
exercise is included to show the use of this index in the
activities section (Chapter 3) of this guide. Others may
be found in some of the references listed at the end of this
section.
The sequential diversity index is calculated by dividing the
number of runs by the number of specimens as show below:
Diversity index = number of runs
number of specimens
A-127
-------
Appendix I
A run is a set of like individuals picked up or observed
respectively. A run ends when an individual of another
kind is found. Therefore, a run can consist of only one
individual. The specimen being observed need only be com-
pared with the previous one. If it appears to be similar,
it is part of the same run; if not, it is part of a new run.
The more runs for a given number of specimens, the greater
the diversity.
Suppose that an investigator is observing cellular algae on
a haemocytometer. As he scans the field of view from left
to right, he observes the following: 1st, five cells of the
same kind which he calls type S; 2nd, ten cells of another
kind which he calls type I; 3rd, five cells of type S; and,
finally, one cell of type U. At this point, the investigator
has four runs and 21 specimens. The usual procedure would be
to continue in a like manner until 200 specimens have been
counted; then the diversity index would be computed.
Macroinvertebrates may be poured out onto a grid of some
sort and treated in the same manner. Alternatively, they may
be sorted according to kind. For example, there might have
been 80 green, worm-like specimens, 40 snails, and 80 leeches.
The green, worm-like specimens would be represented by
numbers 1-80, the snails would be 81-120, and the leeches 121-
200. Numbered slips of paper would then be randomly drawn
to determine the number of runs.
If more than 200 macroinvertebrates are in a sample, randomly
pick and sort until 200 specimens have been removed. The
remaining organisms can be discarded. (If results from
different locations are to be meaningfully compared, the
diversity indices should be computed for the same number of
specimens.)
5. The Significance and Interpretation of Biological Data
The interpretation of data is a time-consuming process.
Biological variability often confuses the beginner. One of the
commonest examples of this is that a few individuals of the same
species will respond differently than the others to apparently
the same environmental conditions. One should be aware of
exceptions, but he should not be disturbed if he cannot explain
them.
In order to get the most complete picture of an aquatic system,
data from as many parameters as possible should be studied. As
a matter of fact, data from a single parameter may mean nothing
by themselves. Finding interrelationships among the various
parameters and relating these to the whole are extremely
important.
A-128
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Appendix I
Care should be exercised to be sure that the data were collected
from a representative sample. Following are a series of tables
which should aid in the interpretation of data. The information
contained in them is not absolute; it should be used only as a
guide.
Warning
If the relationships do not seem to apply to your
investigation, other factors (perhaps requiring equipment
not available to you) may be involved. Rather than risk
publicizing an unwarranted conclusion, seek the advice of
a professional.
Cross-check related observations in different tables.
For example, if low DO is detected under chemical testing,
check turbidity under physical observations and severe
organic pollution under biological observations.
The interpretation of phrases such as "great variety,"
"less variety," and "high coliform count" may pose a problem
for beginners. Most states have developed water quality
standards which will be helpful in the interpretation of
bacterial and some chemical data. If professional macro-
invertebrate data (or assistance) are not available to the
beginning investigator, he will have to collect extensive
data himself; then make his own interpretations.
Table 1 - C - 1 Biological Observations
In Case of: Look for or Expect:
1. Using Sequential Diversity Index (SDI)
Great variety with few of each kind Clean water
Less variety with great abundance Overly enriched
(Moderate organic
pollution)
One or two kinds only, with very great Severe organic
abundance pollution
A-129
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Appendix I
In Case of: Look for or Expect:
2. The Qualitative Interpretation of Freshwater
Macroi nvertebrates
May fly, caddis fly, and stone fly Clean water
larvae, plus a considerable variety
of other macroinvertebrates
Pollution tolerant types predominate, Moderate organic
although a few less tolerant or unknown pollution
forms may be present Suggestion: Confirm with
coliform and other tests.
One or two pollution tolerant types only, Severe organic pollution
often present in overwhelming abundance Suggestion: Same as
above.
No macroinvertebrates at all, little or Toxic pollution
no plant life Suggestion: Same as
above.
3. Quantitative Interpretation of Freshwater Invertebrates
from Riffle Areas
Note: Carefully review weather records for the preceding
few weeks. A severe flood could invalidate the
following interpretations.
0-2 grams per ft.2 (blotted Unproductive, probably
live weight) clean stream.
Suggestion: Check for
toxicity.
3-5 grams per ft.2 Normally productive.
Probably well balanced
stream community.
Over 6 grams per ft.2 A. Highly productive
stream, probably
organically enriched
(polluted).
A-130
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Appendix I
In Case of:
Look for or Expect:
4. Productivity Measurements
Water at sampling site seems to be un-
productive or overproductive
5. Fish Behavior
Many fish observed to be "topping"
(gulping air and/or splashing on surface)
B. Note: These
relative values will
differ in different parts
of the country.
C. Check the SDI.
Suggestion: Measure
plankton productivity
using standing crop,
oxygen, pH, or carbon-14
method (or combination).
Suggestion:
1. Check DO, IDOD, and
BOD.
2. Check for toxic or
oxygen demanding
chemicals.
3. Determine organic con-
tent of water and
bottom debris and/or
sediments.
4. Check temperature.
Table 1 - C - 2 Bacteriological Observations
In Case of:
Look for or Expect:
High coli form count
A. Raw or unchlorinated sewage dis-
charge.
B. Pasture or feed lot drainage.
A-131
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Appendix I
In Case of:
Look for or Expect:
Low coliform count
C. Storm sewer drainage immediately
after a rain storm.
A. Clean water.
B. Heavily chlorinated sewage
effluent.
C. Toxic discharge as from
pharmaceutical company manufacturing
antibiotics, or toxic chemicals.
D. Other source of toxicity such as
acid mine drainage.
Table 1 - C - 3 Chemical Observations
In Case of:
Look for or Expect:
High DO's (12-30 mg/1) durinq day- A. High biological productivity,
light hours (supersaturation) especially producers (plants).
B. Relatively quiet waters.
C. Chemical interference in oxygen
determination.
Suggestion:
1. Check DO between 2 and 3 a.m.
2. Check DO at 1 or 2 hour inter-
vals around the clock and graph
results.
3. Search for source of excessive
fertility.
4. Examine bottom muds for black
anaerobic foul smelling (^S)
deposits that are overgown by
plants.
A-132
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Appendix I
In Case of
Look for or Expect:
Low DO's (0-4 mg/1) during
daylight hours
Toxic chemicals in general
Toxic or smothering chemicals
which sink to bottom
Floating oil slick
5. Compare DO above and below a dam
or rapids area. Deaeration may be
detected.
A. High organic content, both dis-
solved and suspended solids.
B. High total bacteria and fungus
count.
C. If clear water, look for
anaerobic spring (groundwater).
D. Chemical interference in oxygen
determination.
E. Note water temperature.
Suggestion:
1. Check for coliform bacteria.
2. See "Physical, Low Velocity"
(Suggestion #1).
Reduced biological productivity
may be selective or complete.
No living organisms on or in bottom
materials, but overlying water may
have rich plankton and/or nekton
population.
A. Low DO near surface.
B. Oil coated wharf pilings, floats,
and shore.
C. Dead or dying oil soaked birds
and aquatic mammals.
D. Few or no living organisms on oil
covered surfaces.
If in an estuary or open ocean front
shore, this includes the entire
intertidal zone.
A-133
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Appendix I
In Case of:
Look for or Expect:
High pH (8-11), high alkalinity
(200 or more), high hardness
(300 or more)
Low pH (2-5)
Specific chemical effects
A. High turbidity.
B. High biological productivity.
C. If low biological production, look
for toxicity or biologically
intolerable combination of
chemicals.
D. May be of natural origin.
A. Acid mine drainage or industrial
discharge.
B. Low biological productivity.
C. Low turbidity.
See Table 1-D of Appendix I
Table 1 - C - 4 Physical Observations
In Case of:
Look for or Expect:
High velocity and turbulence
A. DO approximately at saturation
for temperature.
B. Hard bottom, little sediment.
C. Biological organisms adapted to
swift water.
D. Particulate materials kept in
suspension.
E. Bank and bottom scouring
(erosion).
A-134
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Appendix I
In Case of:
Look for or Expect:
Low velocity
High volume of flow:
normal, non-flood conditions
(over 1,000 c.f.s.)
Flood conditions (any stream)
A. DO may be above or below
saturation.
B. Coarse particulates may settle to
bottom.
C. Bottom may be soft.
D. Organisms, if present, may be
burrowers or may crawl freely on
surface.
Suggestions:
1. If turbidity is high, see
High Velocity, Item D.
2. Check for kinds and amounts of
plankton.
A. Note: This is probably a "big"
river.
B. These are difficult and expensive
to study, even for professional
groups.
Suggestions:
1. Examine the water for plankton.
2. Place artificial substrates for
both periphyton and macroinverte-
brates.
3. Carry out chemical and physical
analyses of water.
A. High coliforms count in first few
hours, diminishing as time goes on.
B. "Dumping" of waste holding ponds
by industry.
C. "By-passing" of sewage treatment
plants.
A-135
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Appendix I
In Case of:
Look for or Expect:
Low volume of flow (up to 1,000
c.f.s.)
Heated water discharge
(no chemical pollution assumed
although heated discharges often
contain chlorine or other
chemicals used to kill
biological growths in the
plant. In this case, see also
"Toxic Chemical" section.)
Lakes, reservoirs, and
estuaries
D. See also "physical" High
Velocity, Item D, and Low
Velocity, Item B.
A. Note: Smaller streams from 0 up
to 200 - 300 c.f.s. are generally
most satisfactory for group
studies but much depends on local
circumstances, resources, and
objectives.
B. Most of the analyses described can
be carried out on such a stream.
A. Differences between the biota in or
near the discharge canal or pipe
and that in or around the intake.
B. Artificial substrates may be used.
Suggestions:
1. Make the above comparison winter
and summer, or even better, each
season.
2. Chart the dispersal of the heated
water on the receiving water at
different times of the year,
different wind directions,
different tidal phases, etc. If
available, use depth recording
thermometer and include depth as
well as surface temperature.
Graph your results.
A. Thermal (or other density caused)
stratification.
B. Changes in water level, either
natural or man made.
Suggestions:
1. Practically every suggestion
offered elsewhere for stream or
A-136
-------
Appendix I
In Case of: Look for or Expect:
river pollution studies can be
applied to lakes, reservoirs,
or estuaries, making due
allowance for differences in the
basic nature of the waters.
2. Stratification, seiches, density
currents, tidal currents, and
salinity are additional physical
factors to be considered.
3. Biological procedures are
virtually identical, but while DO
is the same, most of the chemical
methods cited apply to fresh-
water only.
A-137
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Appendix 1
Figure 1 - C - 1 Benthos Responses to Pollution
fl.
I
t
I
I
I
ft.
C.
I
t
I
I
I
0,
_, Number of Kinds
-, Number of Organisms
, Sludge Deposits
A-138
-------
Appendix I
Figure 1 - C - 1
Four Basic Responses of Bottom Animals to Pollution
A. Organic wastes eliminate the sensitive bottom animals and provide
food in the form of sludges for the surviving tolerant forms.
B. Large quantities of decomposing organic wastes eliminate sensitive
bottom animals and the excessive quantities of by-products of
organic decomposition inhibit the tolerant forms; in time, with
natural stream purification, water quality improves so that the
tolerant forms can flourish, utilizing the sludges as food.
C. Toxic materials eliminate the sensitive bottom animals; sludge is
absent and food is restricted to that naturally occurring in the
stream, which limits the number of tolerant surviving forms. Very
toxic materials may eliminate all organisms below a waste source.
D. Organic sludges with toxic materials reduce the number of kinds by
eliminating sensitive forms. Tolerant survivors do not utilize the
organic sludges because the toxicity restricts their growth.
A-139
-------
Appendix I
6. Bibliography
Cairns, J., Jr., ejt a][, "The Sequential Comparison Index—A
Simplified Method for Nonbiologists to Estimate Relative
Differences in Biological Diversity in Stream Pollution
Studies," Journal, Water Pollution Control Federation,
1968, pp. 1607-1613"! This is an excellent article and is
highly recommended.
Clark, W. J., and W. F. Sigler, "Method for Concentrating
Phytoplankton Samples Using Membrane Filters," Journal of
Limnology and Oceanography, 1963, 8 (1): 127-129.
Eddy, S., How to Know the Fresh-Water Fishes, Wm. C. Brown Co.,
Dubuque, la., 1957. It is a fairly good reference which can
easily be carried into the field.
Edmondson, W. T., Fresh-Water Biology, (2nd ed.), John Wiley
and Sons, Inc., New York City, 1959. Considered by many to
be the aquatic biologists' "Bible," it is excellent but
very technical reference.
Foerster, J. W., "A Phyco-Periphyton Collector," Turtox News,
1969, 47-3: 82-84. This article describes a simple, easy-
to-make periphyton collector. Collection is made on glass
microscope slides.
Gardiner, A. C., "Measurement of Phytoplankton Population by the
Pigment Extraction Method," Journal of the Marine Biological
Association, 1943, 25 (4): 739-744.
Leech, T. H. E., The Complete Life History of Grossus Disgustus,
The Annelid Press, Franklin, N. H., 1969. It is a highly
recommended book. The author uses this hardy hirudinean to
explain the sequential biological distention of parameters.
A List of Common and Scientific Names of Fishes from the United
States and Canada, American Fisheries Society, Publication
#2, Washington, D. C., 1960.
Pennak, R. W., Fresh-Water Invertebrates of the United States,
Ronald Press, New York City, 1953.This is a good general
reference but very technical.
Pollison, D. P., and W. M. Craighead, Lehigh River Biological
Investigation, Delaware River Basin Commission, Trenton,
N. J., 1968, pp. 24-25. This is a comprehensive report on
a river study. A biomonitoring technique is described;
the report calls it a bioassay.
A-140
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Appendix I
Needham, J. G., and P. R. Needham, A Guide to the Study of
Fresh-Water Biology, Holden-Day, Inc., San Francisco,
Calif., 1962. It is a good reference book with excellent
drawings of organisms.
Sources of Limnological and Oceanographic Apparatus and Supplies,
American Society of Limnology and Oceanography, Publication^
#2, Washington, D. C., 1961. Many specialized items of
biological collecting equipment are not available from the
usual supply houses. This publication lists the ..uppliers.
Standard Methods for the Examination of Water and Wastewater,
(13th ed.), American Public Health Association, Inc., lew
York City, 1971. This book discusses the physical,
chemical, and biological analytical techniques, bioassays,
and chemical analysis. It has good drawings and keys of
organisms. Every school should have at least one copy.
Wilhm, J. L., "Patterns of Numerical Abundance of Animal
Populations," The American Biology Teacher, March 1969,
pp. 147-150. A diversity index is presented as well as
other means of statistically analyzing biological data.
Wilhm, J. L., and T. C. Dorris, "Biological Parameters for
Water Quality Criteria," Biological Science, June 1968,
18-6: 477-481. A comprehensive discussion of the diversity
index is given.
A-141
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Appendix 1
D. Engineering and Physics
A number of useful tools which relate to the previously discussed
chemical, bacteriological, and biological parameters are used in water
studies. In the Engineering and Physics Section, mapping, flow measure-
ments and computer use are discussed as they apply to water pollution
and water surveys.
Map reading can facilitate water survey planning and provide a better
understanding of the body of water being studied. Flow measurement
provides valuable data in interpreting variations in results from chemi-
cal, bacteriological, and biological samplings. The computer Is a
valuable tool in compiling sampling data. A working knowledge in these
three areas will aid in water studies and the interpretation of
collected data.
1. Mapping
In water surveys, maps provide an invaluable tool for the record-
ing of sampling sites and for providing information on water
sources and posssible points of pollution. For example, in the
initial planning of a river study a topgraphic map is useful for
indicating incoming streams and whether those streams pass
through populated areas. Local sewage pipe line maps, obtained
through the city engineer or department of public works, will
pinpoint sources. Thus, before any field work is done, a fairly
good idea of possible sources of pollution may be obtained through
the use of maps.
When a small area is being studied, such as a pond, a map can be
easily drawn using the plane-table survey method. This method
may be used in grade levels 6-12.
Plane-Table Survey Method
a. Equipment
1) Tape measure
2) Wooden stakes
3) A light weight table or head board with some type of
support (i .e., legs)
4) Paper, pencil, ruler
5) Plumb line or carpenter's level
b. Procedure
Although the following procedure is given for mapping a small
A-142
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Appendix 1
pond, the same procedure may be used to map a parking lot, a
school playground, a woodlot, etc. In mapping areas such as
these, sightings would be taken to existing objects (i.e.,
trees, parked cars, swings) rather than driving stakes.
Before the following procedure is started, the mappers should
reconnoiter the site to become familiar with it.
1)
2)
3)
Determine a base line from the ends (A,B) of which almost
all points on the shore line (C-J) are visible. Place
stakes at point A and B.
Place stakes along the shoreline (C-J) so that they are
visible from points A and B.
Place the table or head board at point A. Using the plumb
line or carpenter's level, make sure the table is hori-
zontal .
4) Tape a piece of paper to the table. At eye level to the
table, line up stake B with a ruler and draw a line toward
the sighting. This line is called the base line.
5) From point A line up the other stakes (C-J) with a ruler
and draw a line along the line of sight.
6) Measure the distance between point A and point B with a
tape measure.
7) On the base line sketched in step 4) place point B accord-
ing to the scale desired. For instance, if the distance
between point A and B is 100 feet, point B could be placed
10 inches from point A on the sketched base line. This
would give a scale of "1 inch equals 10 feet."
8) Move the table to point B and again make sure the table is
horizontal.
A-143
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Appendix 1
9) Align the map so that point A may be seen by placing a ruler
along the base line and sighting along the top of the ruler.
10) As soon as the base line is aligned, sight points C-J and
draw the lines toward the sightings.
11) The lines drawn from point B should intersect those lines
drawn from point A. Darken those points and erase the con-
struction lines.
12) Connect all the points with a continuous line. The map
is now to scale as determined in step 7.
13) Fill the map in with whatever information is pertinent
(e.g., north-bearing direction, stream inlets, houses,
etc.).
References and Resources
Phillips, E.A., Field Ecology, D. C. Heath Co., Boston, 1964.
Welch, P. S., Limnological Methods, McGraw-Hill Book Co., New
York City, 1948.
Topographic maps of any area in the United States may be obtained
through the Geological Survey, Department of the Interior,
Distribution Section, Washington, D. C. State index cir-
culars and a folder describing topographic maps may be
obtained free from the above address.
2. Flow
Flow measurements, the velocity and volume of water in a stream,
are among the more important data collected in a water survey.
The velocity of water movement will determine the types of organisms
living in a particular segment of a stream. Likewise, the velocity
will affect the transport of nutrients and organic food past those
organisms attached to stationary surfaces; the transport of plankton
and benthos as drift, which in turn serve as food for higher organisms;
the transport of silts and sediments; and the addition of dissolved
oxygen through surface aeration.
The volume of water in a stream determines to what extent toxic
substances and bacteria are diluted and, therefore, the immediate
effects of an effluent on a stream's condition. Flow measurements
should be determined whenever chemical, bacteriological, or bio-
logical samplings are taken. Variations in chemical, bacteriologi-
cal, or biological results can often be attributed to flow varia-
tions.
A-144
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Appendix 1
Elaborate apparatus is usually used in flow gauging studies, such
as current meters and weirs. The floatation method of estimating
flow, however, can be achieved with simple equipment by students
in the 7th - 12th grades.
a. Velocity
1) Equipment
a) A floating object (float) (This float is to be carried
along by the water and should be as immune as possible
to air flow and should be as visible as possible. In
deep water, an orange will satisfy the needs. For
smaller streams, smaller floats such as corks or rubber
balls have been found very useful.)
b) Measuring tape or calibrated rope in feet
c) Stopwatch or watch with second hand
d) Boots
2) Procedure
a) It is most difficult to measure flow in slowly moving
waters that are over 4 feet deep. This would mean
that a stream with rapids would be best to use for the
float.
b) Locate two points (parallel to flow) in center of stream,
any measurable distance apart.
c) Measure and record the distance between the two points,
making sure that the area is free of any obstructions
(rocks, garbage, etc.).
d) The moment you place the float at the upstream point,
start timing.
e) Mark the time at the instance the float passes the down-
stream point.
f) Using the formula, V(Velocity) = D(Djstance).compute the
value. T (Time)
Note: Average velocity should be calculated to deter-
mine the true velocity of the stream. It can be con-
cluded that the water of a stream or river will flow
fastest on the surface at the center, and that the
average velocity of a stream will, therefore, be less
than the surface velocity.
A-145
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Appendix 1
g) To obtain the average velocity, multiply the surface
velocity by the constant bottom type factor, 0.9 for
smooth bottomed streams or rivers (sand, clay, etc.)
and 0.8 for rough bottomed streams or rivers (rocks,
debris, etc.).
b. Volume
1) Equipment
a) A meter, yard stick or measuring tape for measuring
the depth and width
b) A slide rule, if desired, for calculating the results
2) Procedure
a) Find the average width in feet of the stream between
the same two points used in velocity measurements by
finding the width at regular intervals between the
points and taking the average of the widths.
b) Determine the average depth in feet of the streams
between the points, a certain number of depths from
one side of the stream to the other. Take the average
of these depths at each interval, and then take the
average of the average depths for the entire distance
between the two points.
c) Compute the average cross section of the stream in
square feet between the two points by multiplying the
average width by the average depth.
d) Compute the volume of flow in second-feet (sec./ft.)
by multiplying the average cross section by the average
velocity in feet per second, obtained from the velocity
calculations. The breakdown of units of measurement
in the calculations is as follows:
(feet)2 x feet = (feet)3 or ft.3/sec.
sec. sec.
When flow measurements are being taken from a stream or
river in which wading would be dangerous or impossible,
a boat may be used or measurements may be taken from a
bridge.
c. References
Mackenthun, Kenneth M., The Practice of Water Pollution Biology,
U. S. Department of the Interior, Washington, D. C., 1969"
This paperback contains only half a page on flow but it is a
good text for aquatic biology.
A-146
-------
Appendix 1
Manual on Water. (3rd ed.), American Society for Testing and
Materials, Philadelphia, Pa., 1969. This text has a chapter
on flow measurement but deals with the equipment of interest
to industries.
Grover, N. C., and Harrington, A. R., Stream_F1ow. Dover Publica-
tions, New York City, 1966. This is a good paperback
dealing, among other things, with methods and instruments
for measuring stream flow.
A-147
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Appendix 1
E. Computer Applications
The volumes of data generated from the activities in this text can be
handled most efficiently by a digital computer. The computer's speed
and accuracy is readily adapted to the field of pollution studies as a
tool for the ecologist. Though not everyone is an accomplished pro-
grammer, programs can be generated in the timeshare "BASIC" language
that can be run by almost everyone.
The following programs are written with the nonprogrammer in mind.
By using the Dartmouth College computer, through a remote teletype
terminal, anyone who can type can access the system, call up the
appropriate program (title known front an annotated catalog), and run
that program to obtain the necessary results from his experiment.
The conversational nature of the Dartmouth "BASIC"1 langugage permits
the programmer to write a "prompting" type of program, so that the user
gets the feeling that he is having a "conversation."
These programs are not restricted to the Dartmouth system alone. With
minor changes, they can be accepted by most timeshare systems, or can
be rewritten in Fortran for nontimeshare systems (although they will
lose their conversational nature). Local conditions will dictate how
the computer can be adapted as a tool for the ecologist.
1. STREAM
STREAM calculates the cross section, velocity and volume of flow
of a stream. Input the site information (site number and loca-
tion), the distance in feet from one depth reading to the next,
the depth at each reading in feet and inches, the number of
velocity trials, the distance in feet and inches of each trial, and
the travel time in seconds for each trial. The method of cal-
culating cross section uses a series of triangular and rectangular
areas, where field accuracy dictates volumetric accuracy. The
output is a summary of cross sectional area in ft. , stream velocity
in ft./sec. and volume of flow in ft.-Vsec. A question is then
asked as to whether or not you want a plot. A "yes" will produce
a plot of the cross section of the stream at the site of study.
A "RUN" and "LIST" follow.
BASIC Programming,(Preliminary 5th ed.), Kemeny and Kurtz, Dartmouth
Press, Hanover, N. H., 1969.
A-148
-------
RUN
STREAM 09 AUG 70 20:39
THIS PROGRAM CALCULATES CROSSECT ION., VELOC ITY, FLOW VOLUME
AND PLOTS THE CROSSECTIONAL PROFILE.
DIRECTIONS: ANSWER COMPUTER QUESTIONS.
STREAM CROSSECTION CALCULATION.
SITE NO.? 3
LOCATION? WINNESQUAM RIVER
WIDTH OF STREAM (F,I>? 50,0
HOW MANY DEPTH READINGS WERE TAKEN? 13
DISTANCE FROM SHORE TO FIRST MEASUREMENT(FT.) AND DEPTH (F,I)? 5,2,0
DISTANCE(F),DEPTH(F,I>? 3,3*0
DISTANCE(F),DEPTH(F,I>? 2,4,0
DISTANCECF),DEPTHCF,I>? 2,4,0
DISTANCECF),DEPTH(F, I)? 3,5,0
DISTANCE(F),DEPTH(F,I)? 2,6,0
DISTANCE(F),DEPTH(F,I)? 3,7,0
DISTANCE(F),DEPTHCF,I)? 4,5,0
DISTANCE(F),DEPTH(F,I)? 3,4,0
DISTANCE(F),DEPTH(F,I)? 3,3,0
DISTANCE(F),DEPTH(F,I)? 5,2,0
DISTANCE(F),DEPTH(F,I>? 5,1,0
DISTANCE(F),DEPTH(F, I)? 5,1,0
DISTANCE FROM LAST DEPTH TO SHORE? 5
AVERAGE VELOCITY CALCULATION.
HOW MANY TRIALS WERE CONDUCTED? 2
DISTANCE BETWEEN POINTS(FT.,IN.)? 48,0
TIME OF FLOATCSEC.)? 16.2
DISTANCE BETWEEN POINTS(FT.,IN.)? 50,0
TIME OF FLOAT(SEC.)? 17.5
WAS THE STREAM 1)SMOOTH OR 2) ROUGH BOTTOMED? 1
DATA FOR:WINNESQUAM RIVER
THE AVERAGE VELOCITY OF STREAM AT SITE 3 IS 2.62 FT./SEC.
THE CROSSECTION IS 147 SQ.FT.
THE VOLUME OF FLOW IS 385.1 FT.T3/SEC.
DO YOU WISH A PLOT? YES
A-149
-------
CROSSECTION AT SITE 3 ON WINNESQUAM RIVER
MEASUREMENT WATER LEVEL
BANK *
•
•
•
•
5(2) * .
•
•
8(3) * .
•
10 ( 4 ) *
•
12 ( A ) *
»
•
15 ( 5 ) *
•
17 ( 6 ) *
•
•
20 ( 7 ) *
*
*
•
24 ( 5 ) *
*
•
27 ( 4 ) *
*
*
30 ( 3 ) *
•
•
*
•
35 ( 2 ) * .
•
»
•
•
40 ( 1 ) *.
•
•
•
•
45 ( 1 ) *.
•
•
•
•
BANK ( 50 ) *
DO YOU HAVE ANOTHER SITE TO CALCULATE? NO
A-150
-------
-1-
STREAM
100'
110' CALCULATES CROSSECTION^VELOCITY*FLO¥ VOLUME AND PLOTS SITE PROFILE.
1201
130 PRINT
140 PRINT
150 PRINT "THIS PROGRAM CALCULATES CROSSECTION,VELOCITY,FLOW VOLUME "
155 PRINT "AND PLOTS THE CROSSECTIONAL PROFILE."
170 PRINT
180 PRINT "DIRECTIONS: ANSWER COMPUTER QUESTIONS."
190 PRINT
200 PRINT TAB(15);"STREAM CROSSECTION CALCULATION."
210 PRINT
220 PRINT "SITE NO.";
230 INPUT SI
240 PRINT "LOCATION";
250 INPUT LS
260 PRINT "WIDTH OF STREAM (F,I)";
270 INPUT W,W5
280 LET W=W+W5/12
290 PRINT "HOW MANY DEPTH READINGS WERE TAKEN";
300 INPUT R
310 PRINT"DISTANCE FROM SHORE TO FIRST MEASUREMENT(FT.) AND";
320 PRINT " DEPTH (F,I)";
330 INPUT D(l>,H(1>,ICl>
340 LET H(l )=H( 1 )+I < 1 )/12
350 LET W1=D(1)
360 FOR J=2 TO R
370 PRINT "DISTANCE(F),DEPTH(F,I>";
380 DIM D(20),H(20>, 1(20)
390 INPUT D(J),H(J),I(J)
400 LET H(J)=H(J>+I(J)/12
410 LET W1=W1+DCJ>
420 NEXT J
430 PRINT "DISTANCE FROM LAST DEPTH TO SHORE";
440 INPUT D(R+1)
450 LET W1=W1+D(R+1)
460 IF W1=W THEN 540
470 PRINT
480 PRINT "ERROR IN WIDTH MEASUREMENT."
490 PRINT
500 PRINT "DO YOU WISH TO CONTINUE";
510 INPUT RS
520 IF R$="YES" THEN 540
530 STOP
540 LET T=.5*D(1)*H(1>
550 LET W=W1
560 FOR 1=1 TO R-l
570 IF H(I+1)>H(I) THEN 610
580 LET T=T+.5*(H(I)-H(I+l)>*D(I+1)+DCI+l)*H(I+1)
590 NEXT I
A-151
-------
-2-
STREAM (CONTINUED)
600 GO TO 630
610 LET T=T + .5*(H(I+1 >-H(I»*D(I-H)+HCI >*D(I + 1 )
620 GO TO 590
630 LET T=T+.5*D(R+1)*H(R)
640'VELOCITY
650 PRINT
660 PRINT TAB(15>;"AVERAGE VELOCITY CALCULATION."
670 PRINT
680 PRINT "HOW MANY TRIALS WERE CONDUCTED";
690 INPUT K
700 LET V=0
710 FOR J=l TO K
720 PRINT "DISTANCE BETWEEN POINTS(FT.,IN.>";
730 INPUT F(J),I(J)
740 LET F( J)=F( J)+I ( J)/12
750 PRINT "TIME OF FLOAT(SEC.)";
760 INPUT S(J)
770 LET V=F(J)/S(J)+V
780 NEXT J
790 LET V=V/K
800 PRINT "WAS THE STREAM 1)SMOOTH OR 2) ROUGH BOTTOMED";
810 INPUT A
820 IF A=2 THEN 850
830 LET V=V*.9
840 GO TO 860
850 LET V=V*.8
860 PRINT
870 PRINT "DATA FOR:";L$
880 PRINT
885 LET V=INT(100*V+.5)/100
890 PRINT "THE AVERAGE VELOCITY OF STREAM AT SITE";S1;" IS
900 PRINT V;"FT./SEC."
910 PRINT
915 LET T=INT(100*T+.5)/100
920 PRINT "THE CROSSECTION IS ";T;" SQ.FT."
930 PRINT
935 LET F=INT(10*T*V+.5>/10
940 PRINT "THE VOLUME OF FLOW IS ";F;" FT.T3/SEC."
950 PRINT
960 PRINT " DO YOU WISH A PLOT";
970 INPUT RS
980 IF R$="YES" THEN 1000
990 STOP
1000 GOSUB 1060
1010 PRINT
1020 PRINT"DO YOU HAVE ANOTHER SITE TO CALCULATE";
1030 INPUT RS
1040 IF R$="YES" THEN 190
1050 STOP
1060'PLOT
A-152
-------
-3-
STREAM (CONTINUED)
1070 PRINT
1080 PRINT
1090 PRINT
1100 PRINT TABC10>;"CROSSECTION AT SITE ";Sl;" ON ";L$
1110 PRINT
1120 PRINT "MEASUREMENT";TAB(29);"WATER LEVEL"
1130 PRINT
1140 PRINT "BANK";TAB(34);"*"
1150 LET J=l
1160 LET D9=D(J)
1170 FOR X=l TO W-l
1180 IF X=D9 THEN 1210
1190 PRINT TAB(34);"."
1200 GO TO 1250
1210 IF H(J)<1 THEN 1270
1220 PRINT X;TAB(5);"(";H(J);")";TAB(34-INT(H(J)+.5));"*";TAB(34);"."
1230 LET J=J+1
1240 LET D9=D9+D(J)
1250 NEXT X
1260 GO TO 1290
1270 PRINT X;TAB(33);"*."
1280 GO TO 1230
1290 PRINT "BANK (";W;")";TAB(34);"*"
1300 RETURN
1310 END
A-153
-------
Appendix 1
2. DIV
DIV calculates the diversity index of a sample in a biotic com-
munity using the information and formulas presented in the article
"Biological Parameters for Water Quality Criteria" by Jerry L.
Wilhm and Troy C. Dorris presented in Bioscience, Vol. 18, No. 6.
The diversity index is an indication of pollution levels, for
"values less than 1 have been obtained in areas of heavy pollution,
values from 1 to 3 in areas of moderate pollution, and values ex-
ceeding 3 in clean water areas." Refer to Activity E, Chapter 3.
This program is not conversational in nature. Data must be inserted
as "DATA" statements beginning with line 720. These "DATA" state-
ments must be of the following form: 146, 5, 131, 7, 4, 1, 3, 1,
1.86, etc., where 146 is the total number of individuals, 5 the
number of individual types, 131 through 3 the numbers per individual
population, 1 the site number, and 1.86 the biomass of that sample.
The output is a table listing the individual diversity, diversity
index, theoretical maximum and minimum diversity, and a redundancy
factor. Redundancy expresses the dominance of a type, while
diversity shows the compositional richness of a mixed population
aggregation of organisms.
A "RUN" and "LIST" follows.
"Biological Parameters for Water Quality Criteria," Wilhm and Dorris,
Bioscience, Vol. 18, No. 6, 1968.
A-154
-------
RUN
DIV
09 AUG 70
19:50
INDIVIDUAL
LOCATION DIVERSITY DIVERSITY
1
2
3
4
5
6
9
10
11
12
13
14
15
16
17
18
19
20
21
22
3.94323
4.15551
4.3029
3.74413
4.96296
5.38642
4.2596
3.07844
3.93598
3.22023
4.79031
2.79423
4.92
3.15433
5.21978
2.05037
5.24932
3.46477
4.53496
3.31442
0.657052
1 .34916
0.777571
0.49188
1 .01232
5.36488 E
1 .59888
1 .27176
0.583135
1 .01045
0.730937
1.11113
0.71 1892
0.820883
0.259188
1 .80127
0.20262
1 .45072
0.830485
1 .62428
MAXIMUM
DIVERSITY
2.26021
2.27043
2.44827
1.87177
2.51868
-2 1.97682
2.94719
2.25769
2.19182
2.38556
1.94339
1.82516
2.50029
1 .86566
2.73366
1 .8543
2.55626
2.3364
2.49195
2.46072
MINIMUM
DIVERSITY BIOMASS R
0.196572
0.165477
0.184772
0.17351 1
0.107735
4.63947 E-2
0.30109
0.465142
0.198744
0.425624
7.50793 E-2
0.347148
0.1 12172
0.267617
0. 105908
0.748345
8.62842 E-2
0.346989
0.152622
0.466243
1 .86
1.05
1.5
0.2
0 0.
6.1
2.3
0. 15
0.64
0.67
0.9
2.36
5.48
0.85
5.4
1 .8
6.0
1 .5
1.8
1 .5
0.776861
0.437667
0.738105
0.812532
624798
0.996242
0.509546
0.550017
0.807137
0.70161
1 0.648957
0.483105
0.748873
0.653786
0.941669
0.04795
1 0.9529
0.445195
0.710232
0.419378
R IS A REDUNANCY EXPRESSION WHICH IS THE
DOMINANCE OF ONE OR MORE SPECIES AMD IS THE
INVERSE PROPORTION TO THE WEALTH OF THE SPECIES.
A-155
-------
DIV
100 REM SOURCE:BIOLOGICAL PARAMETERS FOR WATER QUALITY CRITERIA
110 REM BY WILHM AND DORRIS FROM BIOSCIENCE VOL.18 NO.6
120 REM
130 REM PREPARED BY WPP - TILTON SUMMER '70
140 REM
150 PRINT TAB(10>;"INDIVIDUAL"JTAB(32>;"MAXIMUM";TAB(43>;"MINIMUM"
160 PRINT "LOCATION";TAB(10>;"DIVERSITY";TAB<21);"DIVERSITY";TAB(32)
170 PRINT "DIVERSITY";TABC43).;"DIVERSITY";TABC54>;"BIOMASS R"
180 PRINT
190 PRINT
200 PRINT
210 GOSUB 310
220 GOSUB 460
230 GOSUB 530
240 GOSUB 620
250 READ 0>R
260 IF 0<10 THEN 700
270 PRINT TAB(2);0;
280 PRINT TAB(10);QjTABC21);E;TABC32>;M(1>;TAB(43>;
290 PRINT M(2>;TABC54>;R;/CM<1>-M(2»
300 GOTO 200
310 DIM N(72),A(50)
320 READ Z
330 IF Z = 0 THEN 840
340 LET N=LOG(Z)
350 FOR T=Z-l TO 1 STEP -1
360 LET N=LOGCZ-T)+N
370 NEXT T
380 READ S
390 LET B=0
400 FOR C=l TO S
410 READ A CO
420 LET B=LOGCACC))+B
430 NEXT C
440 LET Q=(1/Z)*(N-B)
450 RETURN
460 DIM E(72)
470 LET D=0
480 FOR 1=1 TO S
490 LET D=CACI>/Z)*LOG/Z)/LOGC2)+D
500 NEXT I
510 LET E=-D
520 RETURN
530 LET F=N/LOG(2)
540 LET G=lNTCZ/S+.5)
550 LET H=LOG(G)
560 FOR C=G-1 TO 1 STEP -1
570 LET H=LOG CO+H
580 NEXT C
590 LET G=H/LOG<2)
A-156
-------
-2-
DIV (CONTINUED)
600 LET M(l>=(F-S*G>/Z
610 RETURN
620 LET J=Z-CS-1)
630 LET K=LOGCJ>
640 FOR C = J-1 TO 1 STEP -1
650 LET K=LOGCC)+K
660 NEXT C
670 LET K=K/LOG(2)
680 LET MC2)=(F-K)/Z
690 RETURN
700 PRINT TAB(3);0;
710 GO TO 280
720 DATA 146,5*131*7,4,1,3*1,1.86*181*5,99,68*1*1*12,2, 1.05*208
730 DATA 6,180,1*18*3,1,5,3,1.5,119,4,110,4*1*4,4*.2,401*6,326
740 DATA 17,9,1,6,42,5,0,596*4,593,1,1,1,6,6.1,203,9,118,2,61
750 DATA 11*2,1,1,6,1,9,2.3,64,6,48,2*9,2,2,1,10,.15,144,5,131
760 DATA 6,5,1,1,11,.64,72,6*59,7, 1, 1
770 DATA 3,1,12,.67,335,4,288,15,31,1,13,.91,48*4,35,1,2,10
780 DATA 14*2.36,382*6,336,5,30,1,8*2,15*5.48*68,4,58,2,3,5
790 DATA 16,0.85,509,7,494,3,2,1,2*6,1,17,5.40,24,5*8,1,3
800 DATA 1, 11,18,1 .8
810 DATA 523,6,511,1,6,1,2,2,19,6.01,94,6,62,3,21,1,2,5,20,1.5
820 DATA 263,6,228,9,13,9*3,1,21,1.8,81,7,3,2,30,41,1,2,1,22,1.5
830 DATA 0,0,0,0*0*0,0,0*0,0,0*0*0,0,0,0,0
840 PRINT
850 PRINT
860 PRINT
870 PRINT
880 PRINT
890 PRINT
900 PRINT "R IS A REDUNANCY EXPRESSION WHICH IS THE"
910 PRINT "DOMINANCE OF ONE OR MORE SPECIES AND IS THE"
920 PRINT "INVERSE PROPORTION TO THE WEALTH OF THE SPECIES."
930 END
A-157
-------
Appendix 1
3. DIVERS
DIVERS is similar to DIV, except it incorporates Stirling's for-
mula, H = ~E (Ni/N) 1og2 (Ni/N)1,to indicate population diversity
where N indicates total population count and Ni indicates count
per individual type. The run time of this program is far less
than DIV, and the printout also gives a comment as to pollution
level as noted under the comment for DIV.
This program is also nonconversational in nature and you must
replace the data using "DATA" statements after line 370. The
form is as follows: 1, 146, 5, 131, 7, 4, 1, 3, etc. Where 1
is the site number, 146 the total population, 5 the number of
individual types, and 131-3 the number of each individual type.
A "RUN" and "LIST" follows.
"Biological Parameters for Water Quality Criteria," Wilhm and Dorris,
Bioscience, Vol. 18, No. 6, 1968.
A-158
-------
RUN
DIVERS
09 AUG 70 19:57
LOCATION DIVERSITY
COMMENT
1
2
3
4
5
6
9
10
11
12
13
14
15
16
17
18
19
20
21
22
0.657052
1 .34916
0.777571
0.49188
1.01232
5.36488 E-2
1 .59888
1 .27176
0.583135
1 . 0 1 04 5
0.730937
1.11113
0.709969
0.820883
0.259188
1 .80127
0.20262
1 .45072
0.830485
1 .62666
HEAVY POLLUTION.
MODERATE POLLUTION-
HEAVY POLLUTION.
HEAVY POLLUTION.
MODERATE POLLUTION.
HEAVY POLLUTION.
MODERATE POLLUTION.
MODERATE POLLUTION-
HEAVY POLLUTION.
MODERATE POLLUTION.
HEAVY POLLUTION.
MODERATE POLLUTION-
HEAVY POLLUTION.
HEAVY POLLUTION.
HEAVY POLLUTION.
MODERATE POLLUTION-
HEAVY POLLUTION.
MODERATE POLLUTION-
HEAVY POLLUTION.
MODERATE POLLUTION-
TIME:
READY
0.168 SEC
A-159
-------
DIVERS
100'
110' SOURCE: BIOLOGICAL PARAMETERS FOR WATER QUALITY
120' WILHM AND DORRIS - BIOSCIENCE VOL. 18 NO. 6
130'
140' DIVERSITY USING STIRLING'S FORMULA
150*
160 PRINT
170 PRINT "LOCATION"; TABC10); "DIVERSITY"; TABC30);
180 PRINT
190 PRINT
200 READ L,N,S
210 IF N=0.0 THEN 490
220 LET H=0.0
230 FOR 1=1 TO S
240 READ N(I >
250 LET H=(N(I)/N) * (LOG (N( I )/N) /LOG (2 ) ) + H
260 NEXT I
270 LET H=-H
280 PRINT TABC2); L; TABC10); H; TAB(26>;
290 IF H<1 THEN 350
300 IF H<3 THEN 330
310 PRINT "CLEAN WATER."
320 GO TO 200
330 PRINT "MODERATE POLLUTION."
340 GO TO 200
350 PRINT "HEAVY POLLUTION."
360 GO TO 200
370
380
390
400
410
420
430
440
450
460
470
480
490
DATA
DATA
DATA
DATA
DATA
DATA
DATA
DATA
DATA
DATA
DATA
DATA
END
1
3
5
9
1
1
1
1
1
1
2
9
»
J
9
0
1
3
5
146,
208*
401*
203,
,64,
, 144
,335
,381
7,509
9
1
0*
,523
,263
0*0
5
6
6
9
6
*
,
,
,
,
,
, 131,
, 180*
,326,
, 118,
,48,2
5,131
7
1
1
2
,
*
4,288,
6,336
7,494
6,511
6,228
,
,
,
,
,4, 1,3, 2, 181, 5, 99, 68, 1,1,
,18*3*1*5*4*119*4, 110, 4*1
7*9,1,6,42,6,596,4,593, 1,
,61,1 1,2, 1, 1*6* 1
9*2*1*2
5*6*1* 1*12,72,6,59,7, 1, 1*
15*31* 1* 14, 48*4*35, 1,2, 10
5*30, 1*8*2, 16* 68, 4, 58, 2, 3
3, 2, 1,2, 6, 1,18, 24, 5*8* 1,3
1,6,1,2,2,20,94,6,62,3,21
9*13,9*3*1*22*80*7*3*2*30
1
»
1
3
*
*
*
*
2
4
* 1
* 1
5
1*11
1,2*
41* 1
CRITERIA
"COMMENT"
5
* £J* 1
A-160
-------
Appendix 1
4. DPLfDT
DPL0T is a teletype plotting program for data. Some programming
changes are necessary to alter the scale (at present, the program
is set from 0 to 3). Since 60 is a reasonable teletype size (70
is maximum), a factor must be devised by which to multiply the
result in line 270 so as to equal 60 (i.e., for a high scale of
3, multiply by 20). It is necessary to change line 390 to indicate
the new scale and to divide the result (namely R(I)) by the
appropriate multiple in lines 510 and 560.
Input location, name of test, date, remarks (if any), site number,
and result at that site. The output will be a scaled graph on which
the site numbers and results at each site will be printed.
A "RUN" and "LIST" follows.
A-161
-------
OLD DPLOT
READY
RUN
DPLOT
09 AUG 70 19:58
LOCATION ? NESHEMINY
TEST RESULTS FOR ? DIVERSITY
DATE TAKEN ? 6-24-69
REMARKS? COMPLETE WATERSHED
NO. OF SITES TESTED ? 20
SITE NO. AND RESULTS? 1*0.6571
SITE NO. AND RESULTS? 2,1.3492
SITE NO. AND RESULTS? 3,0.7776
SITE NO. AND RESULTS? 4,0.4919
SITE NO. AND RESULTS? 5,1.0123
SITE NO. AND RESULTS? 6,0.05365
SITE NO. AND RESULTS? 9,1.5988
SITE NO. AND RESULTS? 10,1.2718
SITE NO. AND RESULTS? 11,0.5831
SITE NO. AND RESULTS? 12,1.0105
SITE NO. AND RESULTS? 13,0.7309
SITE NO. AND RESULTS? 14,1.1111
SITE NO. AND RESULTS? 15,0.7099
SITE NO. AND RESULTS? 16,0.8209
SITE NO. AND RESULTS? 17,0.2592
SITE NO. AND RESULTS? 18,1.8013
SITE NO. AND RESULTS? 19,0.2026
SITE NO. AND RESULTS? 20,1.4507
SITE NO. AND RESULTS? 21,0.8305
SITE NO. AND RESULTS? 22,1.6266
A-162
-------
TEST RESULTS FOR DIVERSITY TAKEN ON 6-24-69 ON NESHEMINY
REMARKS: COMPLETE WATERSHED
SCALE FROM 0 TO 3.
+ + > • • >
.
1 . *( 0.6571 )
•
2 . *( 1.3492 )
•
3 . *C 0.7776 )
•
4 . *( 0.4919 )
•
5 . *( 1.0123 )
•
6 .*( 0.05365 )
•
9 . *( 1.5988 )
a
10 . *( 1.2718 )
•
11 . *(. 0.5831 )
•
12 . *( 1.0105 )
•
13 . *( 0.7309 )
•
14 *C 1 .1 111 )
•
15 . *( 0.7099 )
•
16 . *( 0.8209 )
•
17 . *C 0.2592 )
•
18 . *( 1.8013 )
•
19 . *( 0.2026 )
*
20 . *< 1.4507 )
»
21 . *( 0.8305 )
•
22 . *( 1.6266 )
TIME: 0.330 SEC.
READY
A-163
-------
-1-
DPLOT
110' PLOT ROUTINE FOR DATA
120'
130 DIM S(30),R<30>
140 PRINT "LOCATION ";
150 INPUT L$
160 PRINT "TEST RESULTS FOR ";
170 INPUT T$
180 PRINT "DATE TAKEN "',
190 IMPUT D$
200 PRINT "REMARKS";
210 INPUT RS
220 PRINT "NO. OF SITES TESTED ";
230 INPUT N
240 FOR 1=1 TO N
250 PRINT "SITE NO. AND RESULTS";
260 INPUT SCI )*R(I )
270 LET R(I )=R(I )*20
280 NEXT I
290 PRINT
300 PRINT
310 PRINT
320 PRINT
330 PRINT "TEST RESULTS FOR ";T$;" TAKEN ON ";D$;" ON ";L$
340 PRINT
350 PRINT "REMARKS: ";R$
360 PRINT
370 PRINT
380 PRINT TABC5);
390 PRINT "SCALE FROM 0 TO 3."
400 PRINT
410 PRINT TAB(5);
420 FOR 1=1 TO 61
430 IF 1=21 THEN 580
440 IF 1=41 THEN 580
450 PRINT ".";
460 NEXT I
470 PRINT
480 FOR 1=1 TO N
490 PRINT TABC5);"."
500 IF RCI)=0 THEN 560
510 PRINT SCI);TAB(5);".";TAB(R(I)+5);"*";"(";R(I)/20;")"
520 NEXT I
530 PRINT
540 PRINT
550 GO TO 600
560 PRINT S(I );TAB(5>;"*";TAB(9);"C";R(I>/20;">"
570 GO TO 520
580 PRINT "+";
590 GO TO 460
600 END
A-164
-------
Appendix 1
ANALYZE
ANALYZE is programmed to help analyze a sample of water quickly
and accurately. It uses as its parameters: pH, C0?> D0> ID0D,
B0D, C0LIF0RM, FECAL C0LIF0RM, FECAL STREP, and PH0SPHATE.
Input location, site number, date, and the above parametric read-
ings obtained from your tests. The output consists of general
comments about each parametric result and, in some cases, suggests
other tests that might be done to obtain a more complete analysis.
A "RUN" and "LIST" follows.
A-165
-------
RUN
ANALYZE 09 AUG 70 20:57
THIS PROGRAM IS DESIGNED TO HELP YOU ANALYZE YOUR SAMPLE.
LOCATION? WINNESQUAM RIVER
SITE #? 3
DATE? 3-24-70
DO YOU HAVE A PH READING? YES
READING? 6.2
DO YOU HAVE A C02 READING? YES
READING? 18
DO YOU HAVE A DISSOLVED OXYGEN CD.O.) READING? YES
READING? 4
TEMP IN C? 13
DO YOU HAVE A READING OF IMMEDIATE DISSOLVED OXYGEN
DEMAND (I.D.O.D.)? YES
READING? 2
DO YOU HAVE A READING OF BIOLOGICAL OXYGEN DEMAND (B.O.D.)? YES
READING? 10
DO YOU HAVE A COLI FORM COUNT PER 100 ML«? YES
READING? 33766
DO YOU HAVE A READING OF FECAL COLIFORM PER 100 ML.? YES
READING? 250
DO YOU HAVE A FECAL STREP READING? YES
READING? 100
DO YOU HAVE A PHOSPHATES READING IN PPM.? YES
READING? 0.02
A-166
-------
RESULTS FROM WINNESQUAM RIVER TAKEN ON 3-24-70 AT SITE 3 .
THE WATER IS NEUTRAL.
C02 COULD BE A LIMITING FACTOR IF D.O. IS LOW AND
PH IS NOT 'NATURAL'.
D.O. IS LOW FOR CLASS 'A' WATERS.
AT 13 DEGREES C.*THE THEORETICAL D.O. SATURATION LEVEL IS
10.6 .
THE PERCENT OF D.O. IN RELATION TO THE THEORETICAL
D.O. SATURATION IS 27 %.
THE 02 BALANCE IN THIS WATER IS POOR. CHECK I.D.O.D.*
B.O.D.*COLIFORM COUNT FOR POSSIBLE CLUES AS TO THE REASON.
NOTE: DISSO=D.O.-I.D.O.D.
DISSO OF 2 SHOWS OXYGEN DEMANDING MATERIAL IN THE WATER.
THE READING OF 10 FOR B.O.D. HAS NO DISTINCT
RELATIONSHIP TO D.O. BECAUSE OF A VOLUMETRIC DIFFERENCE.
TOTAL COLIFORMS ARE CONSIDERED 'RELIABLE' INDICATORS
AS TO THE POSSIBLE PRESENCE OF BACTERIAL PATHOGENS...
SINCE THE TOTAL COUNT IS 33766 PER 100ML- -.THIS BODY
OF WATER IS UNFIT FOR HUMAN CONTACT.
THE PRESENCE OF FECAL COLIFORMS IN WATER INDICATES
RECENT FECAL CONTAMINATION. SINCE THE FECAL COUNT WAS
REPORTED AS 250 PER 100 ML.* THIS BODY OF WATER IS
UNFIT FOR PUBLIC WATER SUPPLY.
SINCE FECAL STREP EQUALS 100 THE CONTAMINATION
IS LIKELY HUMAN WASTE.
PHOSPHATES ARE PRESENT IN SUFFICIENT AMOUNTS, 0.02
PPM* THAT COULD 'TRIGGER' AN ALGAL BLOOM*IF OTHER
CONDITIONS ARE RIGHT.
TIME: 0.998 SEC.
READY
A-167
-------
-1-
ANALYZE
140
150
160
170
180
190
200
210
220
230
240
250
260
320
330
340
350
360
370
380
390
420
430
440
450
460
470
480
490
500
510
520
WATER ANALYSIS PROGRAM
"THIS PROGRAM IS DESIGNED TO HELP YOU ANALYZE YOUR SAMPLE."
"LOCATION";
L$
"SITE #";
SI
"DATE";
D$
100
110
120'
130 PRINT
PRINT
PRINT
PRINT
INPUT
PRINT
INPUT
PRINT
INPUT
MARGIN 65
PRINT
PRINT
PRINT
LET 1=0
270 DIM AC20),BC60)
280 PRINT "DO YOU HAVE A PH READING";
290 GOSUB 1490
"DO YOU HAVE A C02 READING";
1490
DO YOU HAVE A DISSOLVED OXYGEN CD.O.) READING
1490
"DO YOU
"DEMAND
1490
"DO YOU HAVE A READING OF BIOLOGICAL OXYGEN DEMAND
1490
"DO YOU HAVE A COLIFORM COUNT PER 100 ML.";
1490
"DO YOU HAVE A READING OF FECAL COLIFORM PER 100 ML
1490
"DO YOU HAVE A FECAL STREP READING";
1490
"DO YOU HAVE A PHOSPHATE READING IN PPM.";
1490
REM PH=AC1) C02=AC2)
REM FECAL COLIFORM=AC7)
PRINT
PRINT
"RESULTS FROM ";L$;" TAKEN ON ";D$;" AT SITE ";Si;"
300 PRINT
310 GOSUB
PRINT'
GOSUB
PRINT
PRINT
GOSUB
PRINT
GOSUB
PRINT
400 GOSUB
410 PRINT
GOSUB
PRINT
GOSUB
PRINT
GOSUB
HAVE
CI .D,
A READING
O.D.>";
OF IMMEDIATE DISSOLVED OXYGEN"
CB.O.D.)";
02=AC3) IDOD=AC4) BOD=AC5) COLIFORM=AC6)
FECAL STREP=AC8) PHOSPHATES=AC9>
PRINT
PRINT
530 PRINT
540 IF AC1)=-1 THEN 650
550 IF AC1)>8.6 THFN 640
560 IF AC1)>8.2 THEN 620
570 IF AC1)>5.999 THEN 600
580 PRINT "THE WATER IS ACIDIC."
590 GO TO 650
A-168
-------
-2-
ANALYZE (CONTINUED)
600
610
620
630
640
650
660
670
680
690
700
710
720
730
740
750
760
770
780
790
800
810
820
830
840
850
860
870
880
890
900
910
920
930
940
950
960
970
980
990
1000
1010
1020
1030
1040
1050
1060
1070
1080
1090
"THE
650
"THE
650
"THE
WATER IS NEUTRAL.
PRINT
GO TO
PRINT
GO TO
PRINT
PRINT
IF AC2)=-1
IF A(2)>25
IF A(2)>15
PRINT "C02
GO TO 770
PRINT
PRINT
GO TO
PRINT
PRINT
PRINT
PRINT
IF A(3)=-l THEN 1290
IF A(3>>8.2 THEN 830
IF AC3)>5 THEN 880
WATER IS SLIGHTLY ALKALINE.
WATER IS ALKALINE,
THEN 770
THEN 740
THEN 710
IS NOT A
LIMITING FACTOR.
BE A LIMITING
•NATURAL*."
FACTOR IF D.O. IS LOW AND
"C02 COULD
"PH IS NOT
770
"C02 IS PROBABLY A LIMITING FACTOR.EXAMINE THE
"AREA WITH A DI-URNAL STUDY FOR POSSIBLE ALGAL
"BLOOM. EXAMINE, ALSO, PH AND D.O. READINGS."
"D.O. IS LOW FOR CLASS 'A' WATERS."
900
"THE D.O. IS HIGHER THAN IS 'NATURAL*.IF THE D.O. IS ";
"GREATER THAN 10, IT MAY INDICATE AN ALGAL BLOOM OR SOME
"'UNNATURAL* CONDITION.IT IS RECOMMENDED THAT A ";
"COMPLETE 02 ANALYSIS BE DONE AT THIS SITE."
900
IS PROBABLY NOT A LIMITING FACTOR AND WILL SUPPORT
FISH LIFE."
D.O.
MOST
PRINT
GO TO
PRINT
PRINT
PRINT
PRINT
GO TO
PRINT
PRINT
PRINT
PRINT
LET 1=0
LET B(I ) = 14.6
FOR 1=1 TO 50
READ BCD
NEXT I
LET P=(A(3)/B(Q))*100
PRINT "AT ";T;" DEGREES C.,THE
PRINT " LEVEL IS ";B(T)i".M
PRINT "THE PERCENT OF D.O. IN
LET P=INT(P+.5>
PRINT "D.O. SATURATION IS";?;1
DATA 14.2,13.8,13.5,13
DATA 10.8,10.6,10.4,10
DATA 8.4,8.2,8.1,7.9,7
DATA 6.8,6.7,6.6,6.5,6
IF P>=75 THEN 1170
IF P>=50 THEN 1130
PRINT
THEORETICAL D.O. SATURATION"
RELATION TO THE THEORETICAL
1,12.8,12.5,12.2,11.9,11.6,11.3,11.1
2,10.0,9.7,9.5,9.4,9.2,9.0,8.8, 8.7,8.
8,7.6,7.5,7.4,7.3,7.2,7.1,7.0,6.9
4,6.3,6.2,6.1,6.0,5.9,5.8,5.7,5.6
A-169
-------
-3-
ANALYZE (CONTINUED)
1100
1110
1120
1130
1140
1150
1160
1170
1180
1190
1200
1210
1220
1230
1240
1250
1260
1270
1280
1290
1300
1310
1320
1330
1340
1350
1360
1370
1380
1390
1400
1410
1420
1430
1440
1450
1460
1470
1480
1490
1500
1510
1520
1530
1540
1550
1560
1570
1580
1590
PRINT "THE 02 BALANCE IN THIS WATER IS POOR. CHECK I.D.O.D.,";
PRINT "B.O.D.,COLIFORM COUNT FOR POSSIBLE CLUES AS TO THE REASON."
GO TO 1290
PRINT
PRINT " THE 02 BALANCE IS FAIR. IT PROBABLY DOES NOT ACT AS A ";
PRINT "LIMITING FACTOR*ESPECIALLY IF THE TEMP IS <15 CENT."
GO TO 1290
PRINT
IF P>i00 THEN 1250
PRINT "THE 02 BALANCE IS GOOD*AND SHOULD INDICATE A HEALTHY ";
PRINT "STREAM. NO ANAEROBIC CONDITIONS SHOULD BE PRESENT IF ";
PRINT "A REPRESENTATIVE SAMPLE WAS TAKEN...CAUTION! CHECK "J
PRINT "FLOW READINGS FOR THEY MAY MASK ACYUAL CONDITIONS."
GO TO 1290
PRINT
PRINT "02 FIGURES INDICATE A 'SUPER-SATURATED* CONDITION.";
PRINT "RECHECK 02 READING. A SPECIES DIVERSITY IS RECOMMENDED ";
PRINT "- ALONG WITH A COMPLETE ALGAL COUNT..."
PRINT
IF A(4)=-l THEN 1440
IF A(4)>A(3) THEN 1430
IF A(4)<.5 THEN 1430
LET D1=A(3)-A(4)
PRINT
PRINT "NOTE: DISSO=D.O.-I.D.O.D."
PRINT
IF Dl+3<0 THEN 1400
PRINT "DISSO OF "JDU"
PRINT "WATER."
GO TO 1440
PRINT "DISSO OF ";Dl;"
PRINT "WATER ARE NOT A FACTOR."
GO TO 1440
GOSUB 1650
IF A(5)=-l THEN 1710
PRINT
PRINT "THE READING OF ";A<5);" FOR B.O.D. HAS NO DISTINCT ";
PRINT "RELATIONSHIP TO D.O. BECAUSE OF A VOLUMETRIC DIFFERENCE."
GO TO 1710
INPUT R$
LET 1=1+1
IF RS="NO" THEN 1620
IF R$="YES" THEN 1580
PRINT "INCORRECT FORMAT, PLEASE TYPE YES OR NO."
PRINT "DO YOU HAVE A READING";
INPUT R£
GO TO 1510
PRINT
PRINT "READING";
INPUT Ad)
SHOWS OXYGEN DEMANDING MATERIAL IN THE
SHOWS OXYGEN DEMANDING MATERIALS IN THE
A-170
-------
-4-
ANALYZE (CONTINUED)
1600
1610
1620
1630
1640
1650
1660
1670
1680
1690
1700
1710
1720
1730
1740
1750
1760
1770
1780
1790
1800
1810
1820
1830
1840
1850
1860
1870
1880
1890
1900
1910
1920
1930
1940
1950
1960
1970
1980
1990
2000
2010
2020
2030
2040
2050
2060
2070
2080
2090
IF 1=3 THEN 1650
GO TO 1630
LET A(I)=-1
PRINT
RETURN
PRINT
INPUT
PRINT
GO TO
PRINT
'TEMP IN C"S
1640
"THESE RESULTS FOR I.D.O.D. ARE IMPOSSIBLE,RECHECK.
RETURN
IF A(6)<=50 THEN 1750
PRINT
IF A(6)<250 THEN 1800
IF A(6)>=250 THEN 1880
IF A(6)=-l THEN 2310
PRINT "THE TOTAL COLIFORM COUNT IS ";A(6)J" PER 100
PRINT " THE COUNT IS SUFFICIENT FOR CLASS 'A* WATER
PRINT
GO TO
PRINT
PRINT
PRINT
PRINT
PRINT
PRINT
PRINT
GO TO
PRINT
PRINT
PRINT
PRINT
PRINT
IF AC7)>0 THEN 2000
IF A(7)=-l THEN 2060
PRINT "THIS BODY OF WATER PROBABLY HAS NO RECENT FECAL ";
PRINT "CONTAMINATION. HOWEVER, IF THE TOTAL COLIFORM COUNT
PRINT "IS >50 PER 100 ML., ANOTHER FECAL COUNT IS ADVISED."
PRINT
GO TO
PRINT
PRINT
PRINT
PRINT
PRINT
GOTO 2210
IF A(9><0.015 THEN 2120
PRINT "PHOSPHATES ARE PRESENT IN SUFFICIENT AMOUNTS,";AC9
PRINT " PPM, THAT COULD 'TRIGGER' AN ALGAL BLOOM,IF OTHER
PRINT "CONDITIONS ARE RIGHT."
ML.
1930
"TOTAL COLIFORM COUNT IS CONSIDERED A 'RELIABLE1 INDICATOR "1
"AS TO THE POSSIBLE PRESENCE OF BACTERIAL PATHOGENS ...";
"THEIR PRESENCE INDICATES INADEQUATE WASTE TREATMENT.";
"SINCE THE TOTAL COUNT IS "jA(6>;" PER 100 ML.* THIS ";
"BODY OF WATER IS LIMITED TO BATHING AND IS UNACCEPTABLE ";
"FOR PUBLIC WATER SUPPLY."
1930
"TOTAL COLIFORMS ARE CONSIDERED 'RELIABLE' INDICATORS ";
"AS TO THE POSSIBLE PRESENCE OF BACTERIAL PATHOGENS...";
"SINCE THE TOTAL COUNT IS ";AC6>;" PER 100ML. ,THIS BODY "J
"OF WATER IS UNFIT FOR HUMAN CONTACT."
2060
"THE PRESENCE OF FECAL COLIFORMS IN WATER INDICATES ";
"RECENT FECAL CONTAMINATION. SINCE THE FECAL COUNT WAS ".
"REPORTED AS ";AC7>;" PER 100 ML., THIS BODY OF WATER IS
"UNFIT FOR PUBLIC WATER SUPPLY."
A-171
-------
-5-
ANALYZE (CONTINUED)
2100
21 10
2120
2130
2140
2150
2160
2170
2180
2190
2200
2210
2220
2230
2240
2250
2260
2270
2280
2290
2300
2310
PRINT
GO TO
PRINT
PRINT
PRINT
PRINT
PRINT
PRINT
PRINT
PRINT
GO TO
IF AC
IF A(
PRINT
PRINT
PRINT
GO TO
PRINT
PRINT
PRINT
GO TO
END
2310
"PHOSPHATES..REPORTED TO BE PRESENT IN QUANTITIES OF ";A(9);
" PPM, SEEM SUFFICIENTLY LOW TO PARTIALLY INHIBIT ";
"THE PRODIGIOUS GROWTHS OF ALGAE. IF ALGAE DOES PERSIST ";
"IN LARGE NUMBERS, CHECK THE FOLLOWING:"
TABC13 >;"NITRATE-NITROGEN"
TAB(13);"NITRITE-NITROGEN"
TAB(13)J"AMMONIA COMPOUNDS"
2310
8)=-l THEN 2060
7)/A(8)>l THEN 2270
"SINCE FECAL STREP EQUALS
"IS LIKELY ANIMAL WASTE."
2060
"SINCE FECAL STREP EQUALS
"IS LIKELY HUMAN WASTE."
2060
';AC8>;" THE CONTAMINATION
';A(8>;f> THE CONTAMINATION
A-172
-------
Appendix 1
6. STR-CLAS
STR-CLAS is a stream classification program using New Hampshire
standards established January 1, 1970.* See Table 1 at end of
this appendix. The parameters for the classification are: pH,
D0, TURBIDITY, HYDR0CARBON (0IL), C0LIF0RM, FECAL C0LIF0RM, AND
FECAL STREP. By inputting the above parameters from any site study,
the output will consist of a classification (A,B,C,D) per para-
meter. The lowest classification being the stream classification
at that site.
A subprogram, "STREAM 1," calculates volume of flow and another
subprogram, "SATTABLE," provides a table of DO saturations from
0° to 50° C for checking your results. A basic description of the
final classification is also available.
A "RUN" and "LIST" follows.
* This program will be revised to use Federal Standards,
A-173
-------
RUN
STR-CLAS 09 AUG 70 20:46
THIS PROGRAM CLASSIFIES WATER PER SITE BY N.H.STANDARDS
LOCATION ? WINNESQUAM RIVER
DATE OF TEST ? 7-24-70
SITE # ? 3
DO YOU HAVE A PH READING? YES
READING? 6.2
DO YOU HAVE A DISSOLVED OXYGEN CD.O.) READING? YES
READING? 4
WATER TEMP IN C? 13
DO YOU HAVE A TURBIDITY READING? YES
READING? 22
DO YOU HAVE A HYDROCARBON (OIL) FACTOR? NO
DO YOU HAVE A COLIFORM COUNT PER ML.? YES
READING? 33766
DO YOU HAVE A FECAL COLIFORM READING? YES
READING? 250
DO YOU HAVE A FECAL STREP READING? YES
READING? 100
DO YOU HAVE A FLOW CALCULATION TO MAKE? NO
DO YOU HAVE THE VOLUMETRIC VALUE ALREADY? YES
VALUE? 385.1
A-174
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SOURCE: NEW HAMPSHIRE WATER SUPPLY AMD POLLUTION CONTROL
COMMISSION. JAN.1,1970.
NOTE: THE FINAL CLASSIFICATION IS THE LOWEST CLASS NOTED BELOW!
RESULTS OF WATER CLASSIFICATION FOR SITE 3 ON WINNESQUAM RIVER FOR
7-24-70
A PH OF 6.2 INDICATES CLASS C WATER.
A D.O. OF 4 INDICATES CLASS D WATER.
A TURBIDITY READING OF 22 INDICATES CLASS B OR C WATER,
COMMENT:NON-TROUT STREAM ACCEPTABILITY.
A TOTAL COLIFORM READING OF 33766 PER 100 ML.
INDICATES CLASS C OR D WATER.
COMMENT:
ANY FECAL COLIFORM OR FECAL STREP READINGS (. YOURS
ARE 250 AND 100 ) INDICATES RECENT
CONTAMINATION BY WARM BLOODED ANIMALS.
THIS CONTAMINATION IS LIKELY HUMAN WASTE.
ALL OF THIS IS BEING CARRIED ALONG AT 385.1 FTt3/SEC.
DO YOU WISH AN EXPLAINATION OF THE WATER CLASSES (YES/NO)? YES
WHICH CLASS OF WATER (A,B,C,D>? C
CLASS C: ACCEPTABLE FOR RECREATIONAL BOAT ING,FISHING,
AND INDUSTRIAL WATER SUPPLY WITH OR WITHOUT TREATMENT*
DEPENDING ON INDIVIDUAL REQUIREMENTS. (. THIRD HIGHEST
QUALITY).
WOULD YOU LIKE ANOTHER CLASS EXPLAINED (YES/NO)? YES
WHICH CLASS OF WATER (A,B,C,D>? D
CLASS D: AESTHETICALLY ACCEPTABLE. SUITABLE FOR CERTAIN
INDUSTRIAL PURPOSES, POWER AND NAVIGATION. (LOWEST ALL-
OWABALE QUALITY NOW LESS THAN 1/2 MILE IN ENTIRE STATE).
WOULD YOU LIKE ANOTHER CLASS EXPLAINED (YES/MO)? NO
DO YOU WISH A TABLE OF THEORETICAL D.O.SATURATION FROM 0 TO 50 CENT.? YE
A-175
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THEORETICAL D.0.SATURATION TABLE FROM 0 TO 50 CENT.
THEORETICAL
TEMP (°C) D.O. (ppm)
0 14.6
1 14.2
2 13.8
3 13.5
4 13.1
5 12.8
6 12.5
7 12.2
8 11.9
9 11.6
10 11.3
11 11.1
12 10.8
13 10.6
14 10.4
15 10.2
16 10
17 9.7
18 9.5
19 9.4
20 9.2
21 9
22 8.8
23 8.7
24 8.5
25 8.4
26 8.2
27 8.1
28 7.9
29 7.8
30 7.6
31 7.5
32 7.4
33 7.3
34 7.2
35 7.1
36 7
37 6.9
38 6.8
39 6.7
40 6-6
41 6.5
42 6.4
43 6.3
44 6.2
45 6.1
46 6
47 5.9
48 5.8
49 5.7
50 5.6
A-176
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-1-
STR-CLAS
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PRINT
SUB STREAM1;SATIABLE
MARGIN 70
I
' WATER CLASSIFICATION PROGRAM FOR NEW HAMPSHIRE.
' WRITTEN DURING WATER POLLUTION PROGRAM - SUMMER '70
PRINT " THIS PROGRAM CLASSIFIES WATER PER SITE BY M.H.STANDARDS."
PRINT
PRINT
PRINT "LOCATION ";
INPUT L$
PRINT "DATE OF TEST "',
INPUT D$
PRINT "SITE # ";
INPUT SI
LET 1=0
PRINT
PRINT "DO YOU HAVE A PH READING";
GOSUB 2510
PRINT "DO YOU HAVE A DISSOLVED OXYGEN (D.O.) READING";
GOSUB 2510
PRINT "DO YOU HAVE A TURBIDITY READING";
GOSUB 2510
PRINT"DO YOU HAVE A HYDROCARBON (OIL) FACTOR";
GOSUB 2510
PRINT "DO YOU HAVE A COLIFORM COUNT PER ML.";
GOSUB 2510
PRINT "DO YOU HAVE A FECAL COLIFORM READING";
GOSUB 2510
PRINT "DO YOU HAVE A FECAL STREP READING";
GOSUB 2510
PRINT "DO YOU HAVE A FLOW CALCULATION TO MAKE";
INPUT R$
IF R$="YES" THEN 510
PRINT "DO YOU HAVE THE VOLUMETRIC VALUE ALREADY";
INPUT AS
IF A$="NO" THEN 530
PRINT "VALUE";
INPUT F
GO TO 530
GOSUB #1
• A(1)=PH A(2)=D.O.
' A(6)=FECAL COLIFORM
t
LET 1=1
IF A(I)=-! THEN 710
IF A(I)>8.5 THEN 640
IF AC I)>8 THEN 660
A(3)=TURBIDITY A(4)=OIL
A(7)=FECAL STREP
A(5)=COLIFORM
A-177
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-2-
STR-CLAS (CONTINUED)
600 IF ACI)>6.5 THEN 680
610 IF ACI)>6 THEN 700
620 GOSUB 2200
630 GO TO 710
640 GOSUB 2200
650 GO TO 710
660 GOSUB 2180
670 GO TO 710
680 GOSUB 2220
690 GO TO 710
700 GOSUB 2180
710 LET 1=2
720 IF ACI)=-1 THEN 870
730 FOR J=0 TO 50
740 READ S(J)
750 NEXT J
760 DATA 14.6*14.2*13.8*13.5*13.1,12.8*12.5*12.2*11.9*11.6*11.3*11
770 DATA 10.8*10.6*10.4*10.2*10.0*9.7*9.5*9.4*9.2*9.0*8.8*8.7*8.5
780 DATA 8.4*8.2*8.1*7.9*7.8*7.6*7.5*7.4*7.3*7.2*7.1*7.0*6.9
790 DATA 6.8*6.7*6.6*6.5*6.4*6.3*6.2*6.1*6.0*5.9*5.8*5.7*5.6*5.5
800 IF ACI )>.75*SCT) THEN 840
810 IF AC I)> 5 THEN 860
820 GOSUB 2200
830 GO TO 870
840 GOSUB 2220
850 GO TO 870
860 GOSUB 2180
870 LET 1=3
880 IF ACI)=-1 THEN 1010
890 IF AC 1X5 THEN 940
900 IF AC 1X10 THEN 960
910 IF ACIX25 THEN 990
920 GOSUB 2200
930 GO TO 1010
940 GOSUB 2140
950 GO TO 1010
960 GOSUB 2240
970 GOSUB 2280
980 GO TO 1010
990 GOSUB 2240
1000 GOSUB 2300
1010 LET 1=4
1020 IF ACI>=-1 THEN 1110
1030 IF ACI)=1 THEN 1080
1040 IF ACI)=2 THEN 1100
1050 PRINT "INPUT EITHER 1 OR 2 FOR THE OIL FACTOR";
1060 INPUT ACI)
1070 GO TO 1030
1080 GOSUB 2180
1090 GO TO 1110
A-178
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-3-
STR-CLAS (CONTINUED)
1100 GOSUB 2200
1110 LET 1=5
1120 IF ACI)=-1 THEN 1200
1130 IF ACI)<50 THEN 1170
1140 IF ACIX240 THEN 1190
1150 GOSUB 2260
1160 GO TO 1200
1170 GOSUB 2140
1180 GO TO 1200
1190 GOSUB 2160
1200 PRINT
1210 PRINT TABC25);
1220 FOR 1=1 TO 10
1230 PRINT "*";
1240 NEXT I
1250 FOR 1=1 TO 4
1260 PRINT
1270 NEXT I
1280 PRINT "SOURCE: NEW HAMPSHIRE WATER SUPPLY AND POLLUTION CONTROL"
1290 PRINT "COMMISSION. JAN.1,1970."
1300 PRINT
1310 PRINT "NOTE: THE FINAL CLASSIFICATION IS THE LOWEST CLASS NOTED";
1320 PRINT " BELOW!"
1330 PRINT
1340 PRINT "RESULTS OF WATER CLASSIFICATION FOR SITE ";S1;" ON ";
1350 PRINT L$; " FOR ";D$
1360 PRINT
1370 PRINT
1380 IF AC1)=-1 THEN 1410
1390 PRINT "A PH OF ";Ad);" INDICATES CLASS ";A$C1);" WATER."
1400 PRINT
1410 IF AC2)=-1 THEN 1440
1420 PRINT "A D.O. OF ";AC2);" INDICATES CLASS ";ASC2);" WATER."
1430 PRINT
1440 IF A(3)=-l THEN 1500
1450 PRINT "A TURBIDITY READING OF ";A(3)J" INDICATES CLASS ";A$C3);
1460 PRINT " WATER."
1470 IF AC3)<10 THEN 2110
1480 IF AC3)<25 THEN 2110
1490 PRINT
1500 IF AC4)=-1 THEN 1590
1510 IF AC4)=1 THEN 1560
1520 PRINT "A DEFINITE HYDROCARBON COIL) OBSERVATION INDICATES CLASS";
1530 PRINT " D WATER."
1540 PRINT
1550 GO TO 1590
1560 PRINT "A SLIGHT HYDROCARBON COIL) FILM INDICATES AT BEST CLASS";
1570 PRINT " C WATER."
1580 PRINT
1590 IF AC5)=-1 THEN 1630
A-179
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-4-
STR-CLAS (CONTINUED)
1600
1610
1620
1630
1640
1650
1660
1670
1680
1690
1700
1710
1720
1730
1740
1750
1760
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1800
1810
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1830
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1880
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1900
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1920
1930
1940
1950
1960
1970
1980
1990
2000
2010
2020
2030
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2050
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2080
2090
PRINT
PRINT
PRINT
IF AC
PRINT
PRINT
PRINT
PRINT
PRINT
IF AC
IF AC
PRINT
GO TO
PRINT
PRINT
PRINT
"A TOTAL COLIFORM READING OF ";AC5);'f PER 100 ML."
"INDICATES CLASS ";ASC5);" WATER."
6) = -l THEN 1740
TABC15);"COMMENT:"
TABC5);"ANY FECAL COLIFORM OR FECAL STREP READINGS C YOURS"
TABC5);"ARE ";AC6);" AND ";A(7)J") INDICATES RECENT"
TABC5);"CONTAMINATION BY WARM BLOODED ANIMALS."
7) = -l THEN 1740
6)/AC7)>l THEN 1730
TABC5);"THIS CONTAMINATION IS LIKELY ANIMAL WASTE."
1740
TABC5);"THIS CONTAMINATION IS LIKELY HUMAN WASTE."
"ALL OF THIS IS BEING CARRIED ALONG AT ";F;" FTt3/SEC."
FOR J=l TO 5
PRINT
NEXT
PRINT
INPUT
IF R$
GO TO
PRINT
PRINT
INPUT
IF L$
IF L$
IF L$
GOSUB
GO TO
PRINT
GOSUB
GO TO
PRINT
GOSUB
GO TO
PRINT
GOSUB
PRINT
PRINT
INPUT
IF R$
PRINT
PRINT
PRINT
INPUT
IF AS
STOP
GOSUB
J
"DO YOU WISH AN EXPLANATION OF THE WATER CLASSES CYES/NO)";
R$
="YES" THEN 1830
2030
"WHICH CLASS OF WATER CA,B,C,D)";
L$
="A" THEN 1910
="B" THEN 1940
="C" THEN 1970
2320
1990
2370
1990
2410
1990
2460
"WOULD YOU LIKE ANOTHER CLASS EXPLAINED CYES/NO)";
R$
="YES" THEN 1840
"DO YOU WISH A TABLE OF THEORETICAL D .0 .SATURATION FROM";
" 0 TO 50 CENT.";
AS
="YES" THEN 2090
#2
A-180
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-5-
STR-CLAS (CONTINUED)
2100 STOP
2110 PRINT
2120 PRINT TAB(5);"COMMENT:";B$(3)
2130 GO TO 1490
2140 LET A$(I)="A"
2150 RETURN
2160 LET A$(I)="B"
2170 RETURN
2180 LET AS(I)="C"
2190 RETURN
2200 LET AS(I)="D"
2210 RETURN
2220 LET A$(I)="A OR B"
2230 RETURN
2240 LET A$(I)="B OR C"
2250 RETURN
2260 LET A$(I)="C OR D"
2270 RETURN
2280 LET B$(I)="TROUT STREAM ACCEPTABILITY."
2290 RETURN
2300 LET B$(I)="NON-TROUT STREAM ACCEPTABILITY."
2310 RETURN
2320 PRINT
2330 PRINT " CLASS D: AESTHETICALLY ACCEPTABLE. SUITABLE FOR CERTAIN"
2340 PRINT " INDUSTRIAL PURPOSES, POWER AND NAVIGATION. (LOWEST ALL-"
2350 PRINT " OWABALE QUALITY NOW LESS THAN 1/2 MILE IN ENTIRE STATE)."
2360 RETURN
2370 PRINT " CLASS A: POTENTIALLY ACCEPTABLE FOR PUBLIC WATER SUPPLY"
2380 PRINT " AFTER DISINFECTION. NO DISCHARGE OF SEWAGE OR OTHER"
2390 PRINT " WASTES. (QUALITY UNIFORMLY EXCELLENT)."
2400 RETURN
2410 PRINT "CLASS B: ACCEPTABLE FOR BATHING AND RECREATION,FISH"
2420 PRINT "HABITAT AND PUBLIC WATER SUPPLY AFTER ADEQUATE TREATMENT."
2430 PRINT " NO DISPOSAL OF SEWAGE OR WASTES UNLESS ADEQUATELY TREATED."
2440 PRINT " (HIGH AESTHETIC VALUE.)"
2450 RETURN
2460 PRINT " CLASS C: ACCEPTABLE FOR RECREATIONAL BOAT ING,FISHING,"
2470 PRINT " AND INDUSTRIAL WATER SUPPLY WITH OR WITHOUT TREATMENT,"
2480 PRINT " DEPENDING ON INDIVIDUAL REQUIREMENTS. ( THIRD HIGHEST"
2490 PRINT " QUALITY)."
2500 RETURN
2510 LET 1=1+1
2520 INPUT R$
2530 IF R$="NO " THEN 2640
2540 IF R$="YES" THEN 2590
2550 PRINT "INCORRECT FORMAT. PLEASE TYPE YES OR NO."
2560 PRINT " DO YOU HAVE A READING";
2570 INPUT R$
2580 GO TO 2530
2590 IF 1=4 THEN 2700
A-181
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-6-
STR-CLAS (CONTINUED)
2600 PRINT "READING";
2610 INPUT A(I)
2620 IF 1=2 THEN 2670
2630 GO TO 2650
2640 LET A(I)=-1
2650 PRINT
2660 RETURN
2670 PRINT "WATER TEMP IN C";
2680 INPUT T
2690 GO TO 2650
2700 PRINT "(1) SLIGHT OIL FILM OR (2) OIL SLICK";
2710 INPUT A(I)
2720 GO TO 2650
2730 END
A-182
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-1-
STREAM1
100 DIM D(20),H(20).» I (20)
110 PRI NT
120 PRINT
130 PRINT TAB(15>;"STREAM CROSSECTION CALCULATION."
140 PRINT
150 PRINT
160 PRINT "WIDTH OF STREAM (F,I>";
170 INPUT W..W5
180 LET W=W+W5/12
190 PRINT "HOW MANY DEPTH READINGS WERE TAKEN";
200 INPUT R
210 PR I NT"DISTANCE FROM SHORE TO FIRST MEASUREMENT(FT.) AND1
220 PRINT " DEPTH (F,I)";
230 INPUT D(l>,H(1>,I( 1 )
240 LET H(l)=H(1)+I(1)/12
250 LET W1=D( 1 )
260 FOR J=2 TO R
270 PRINT "DISTANCE(F),DEPTH(F>I)";
280 INPUT D(J),H(J),I(J)
290 LET H(J)=H(J)+I(J)/12
300 LET W1=W1+D(J)
310 NEXT J
320 PRINT "DISTANCE FROM LAST DEPTH TO SHORE";
330 INPUT D(R-H)
340 LET W1=W1+D(R+1)
350 IF W1=W THEN 430
360 PRINT
370 PRINT "ERROR IN WIDTH MEASUREMENT."
380 PRINT
390 PRINT "DO YOU WISH TO CONTINUE";
400 INPUT R$
410 IF R$="YES" THEN 430
420 STOP
430 LET T=.5*D(1>*H(1>
440 LET W=W1
450 FOR I =1 TO R-l
460 IF H(I+1)>H(I) THEN 500
470 LET T=T+.5*(H(I)-H(I+l))*D(I+1)+D(I+1)*H(I+1>
480 NEXT I
490 GO TO 520
500 LET T=T + .5*(HCI+1)-H(I>>*D(I-H >+H(I )*D(I + 1 >
510 GO TO 480
520 LET T=T+.5*D(R+1)*H(R)
530'VELOCITY
540 PRINT
550 PRINT TAB(15);"AVERAGE VELOCITY CALCULATION."
560 PRINT
570 PRINT "HOW MANY TRIALS WERE CONDUCTED";
580 INPUT K
LET V=0
A-183
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-2-
STREAM1 (CONTINUED)
600 FOR J=l TO K
610 PRINT "DISTANCE BETWEEN POINTSCFT.,IN.)";
620 INPUT FCJ),ICJ>
630 LET F(J)=F(J)+I(J)/12
640 PRINT "TIME OF FLOAT(SEC.)";
650 INPUT SCJ)
660 LET V=F(J)/S(J)+V
670 NEXT J
680 LET V = V/K
690 PRINT "WAS THE STREAM 1)SMOOTH OR 2) ROUGH BOTTOMED";
700 INPUT A
710 IF A = 2 THEN 740
720 LET V=V*.9
730 GO TO 750
740 LET V=V*.8
750 LET F=T*V
760 LET F=INT(F+.5>
770 PRINT
780 PRINT "FOR A MORE COMPLETE ANALYSIS,USE THIS DATA IN 'STREAM'
790 PRINT
800 RETURN
810 END
A-184
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-1-
SATTABLE
100 DIM S(51 )
110 ' THEORETICAL D.O. SATURATION TABLE.
120 PRINT
130 PRINT
140 PRINT "THEORETICAL D.0.SATURATION TABLE FROM 0 TO 50 CENT
150 PRINT
160 PRINT TAB(26>;"THEORETICAL"
170 PRINT TAB(5);"TEMP";TAB(30);"D.O."
180 PRINT
190 FOR 1=0 TO 50
200 IF I>9 THEN 260
210 PR I NT TAB ( 6 ) ', I ',
220 IF SCIX10 THEN 280
230 PRINT TAB(28>;S(I>
240 NEXT I
250 RETURN
260 PRINT TABC5);i;
270 GO TO §20
280 PRINT TAB(29>;SCI>
290 GO TO 240
300 END
A-185
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Appendix 2
Implementation
The implementation of the activities depend on the school's ability to
handle several problems, namely: cost, scheduling, and motivation. These
problems are interrelated at most schools; therefore, they must be resolved
as the program proceeds. Beginning with a small group of interested admin-
istrators, teachers, and students seems to be a good approach. A club or
single course offering may be as far as a school can proceed in the first
year. The schools listed in the preface have programs underway or are
beginning programs. Furthermore, they are grouped into clusters which
meet monthly, carry on interschool activities, and support the implementa-
tion of the program in their region. The organized regions are: Central
New Hampshire, Southern New Hampshire-Vermont, Eastern Massachusetts,
Central Massachusetts, Northern Massachusetts, New York-Pennsylvania-New
Jersey, Washington (D.C.), and South.Carolina-Georgia. Any school listed
in the preface may be contacted to obtain the cluster coordinator's name.
The sections below deal with the various problems of implementation.
A. Cost
The expense involved in beginning a program can be minimized in several
ways; costs are increased by equipment, travel, and books and references.
Equipment for all but a few activities is not specialized and probably
already exists in the various school laboratories. For example, any
of the chemical analyses may be made in the lab. Usually, however, this
makes testing slow, inconvenient, and remote for the test site. Field
test kits which are modular can be obtained to perform selected tests;
an elementary kit which contains tests for DO, temperature, turbidity,
pH, chlorine, phosphate, sulphate, and nitrate are sufficient to test.
Bacterial studies do require equipment which may not be on hand; con-
sult with the staff of your local hospital, county health department,
and sewage treatment plant. Usually an arrangement may be worked out
to use their facilities.
If your school is a nonprofit enterprise it qualifies for U. S. govern-
ment surplus. To obtain information on the location of such surplus
depots, contact your state education department. Local industry has
been found to be particularly helpful; money may not be too easily
obtained, but often slightly used equipment or materials will be made
available. If you make your needs known through news media and at
community meetings and fairs, almost anything can happen. Students at
Germantown Academy have helicopter service available for aerial photo-
graphy as a result of their activities in mapping and monitoring the
Wissahickon Creek.
Much of the equipment listed in the activities may be made by hand. If
students and teachers work together on this project, a great deal of
equipment may be made in a short time. The making of equipment leads to
new and better methods of inquiry.
A-186
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Appendix 2
B. Scheduling
Teachers have noted that informal arrangements with other teachers
allow the greatest flexibility when the school is unable to provide
adequate scheduled time. The sports department arrangements for team
games usually may be duplicated if the importance of academic pursuits
can be established. Past experience indicates that as the school gains
recognition in the community for its achievements, the difficulty in
getting time for activities diminishs rapidly. Activities such as (G)
and (H) in Chapter 4 are good beginning places to arouse community
interest.
C. Motivation
Helping to build the necessary interest ir pollution among teachers,
students, administrators, and parents will be an important role which
you must perform. The pollution problems which we face today are, in
themselves, grave enough to motivate people to action. Therefore, in
many cases, all that needs to be supplied is a vehicle for expressing
and focusing this action. This can very often be accomplished through
a club, the planning and presentation of an assembly program, or a
variation of an earth day program.
Any of the above activities may serve to allow the students, faculty,
administration, and even the parents to become involved and interested
in some particular facet of our pollution problems. Once interest and
activity have begun there are several means of aiding in its continuance.
School administrators are particularly fond of anything which gains
favorable publicity for the school. Most of the above mentioned
activities will produce this in the form of newspaper articles and radio
and television newscasts. All that you need do is be sure to advise
them of what you are planning.
We have found that one factor which aids in the sustained effort of
the students is to make them aware of your belief that they are capable
of making significant contributions to the community at large. They
do possess the initiative and curiosity to determine problems, conduct
research, and translate the information into meaningful conclusions,
but it is important that you make your awareness of this known to them.
A published statement to this effect on the part of the faculty, the
publication of a student journal, or the submission of their data and
conclusions to the appropriate public agencies serves to substantiate
this in the minds of the students.
Since students enjoy this activity-oriented approach and, at the same
time, learn more through it, other teachers will want to know what
you are doing. Do not be afraid to explain it to them or even better,
invite them to participate in the activities along with the students.
Your success should provide the necessary motivation for them to try
it also.
A-187
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Appendix 2
It is wise to motivate as many parents as possible. Naturally, they
are curious about what their children are doing in school. Invite them
to participate or at least arrange for a demonstration for them. They
may prove to be an important resource for you in terms of the informa-
tion, equipment, and transportation which they may be able to provide.
A-188
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Appendix 3
Limitations
There are a number of possible trouble spots which may pop up during
the implementation of these activities. A foreknowledge of these problem
areas will, in most cases, be all that is needed to avoid them. In general
the problems seem to fall into three categories: time and transportation,
methods and equipment, and dealing with other people.
A. Time and Transportation
1. Most activities in thie guide are designed to last an hour and a
half; however, many do require more time for completion. For
one of the filming activities, for example, students camped
overnight on Cardigan Mountain in order to film the sunrise.
2. Some lab experiments required by these activities take a long time
to obtain the results. The BOD test, for example, requires 5 days
before final results can be obtained. Thus it may become a test
of your students' patience.
3. Some people will have difficulty locating suitable streams or lakes
for their activities. Streams with the easily monitored depth,
rate of flow, width, suitable access points, etc., are not always
close to the school.
4. Transporting a large class to and from a site may pose problems
in simply obtaining permission to use school busses and to miss
lunch, parents' permissions, a driver for the school bus, box
lunches, or money for lunches. Methods must be devised to over-
come such obstacles. Often making friends with the janitors,
cafeteria people, and bus drivers is the key.
B. Methods and Equipment
1. After gaining permission to leave school with your class and having
organized all these details, do not forget to advise parents that
the students may be required to wade in streams and grovel in the
dirt. Students' clothes may become somewhat soiled.
2. Also before leaving the school, be sure you have all the equipment
necessary for the activity. This reduces transportation costs and
total time required.
3. Students should be well versed in the care of equipment in the field
since it is much more difficult to keep it dry and clean.
4. When sterile procedures are required in the field, special care must
be taken because the chances of contamination seem to increase in
proportion to the distance from the lab.
A-189
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Appendix 3
5. There is some danger involved when students are dealing with sewage
wastes. Hip boots and rubber gloves are sometimes a necessity.
6. We have found some keys especially difficult to use. Keep looking
for one that is not as vague as the one you are presently using.
7. If the cost of some of the equipment is prohibitive in your case,
please use Appendix 2 on Implementation to help remedy these
difficulties.
C. Dealing with Others
1. When taking relatively large groups to confined places (a city
manager's office, for example) or a privately owned area (such as
a farm), it is advisable that the person or persons being contacted
understand the size and age level of the group beforehand so that
they will be prepared.
2. Always be sure to arrange your appointments well in advance.
3. If you arrange an interview, be sure to list your questions before
you arrive. In general, people are willing to cooperate but their
time is limited.
4. Always gain permission to use private land, even if you are only
using it as access to a lake, river, or stream.
5. Be sure to arrange for the protection of equipment if it must be
left in a certain place for any length of time. Many experiments
have been disrupted or completely destroyed by curious or unknowing
outsiders. Experience has shown that experiments placed in drive-
ways or parking lots are particularly vulnerable.
6. Since these activities are concerned with water pollution, you must
be prepared at some point to deal with the polluters themselves.
To what extent you are willing to incur their wrath is not only a
test of your own moral fortitude but also that of your school.
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Appendix 4
Evaluation
Accountability is an essential ingredient in all educational programs.
Effective fulfillment of this parameter requires careful planning accompanied
by appropriate evaluation. This may seem antagonistic to an approach con-
sisting of activities which are defined only to the point necessary to
initiate thinking processes and promulgated through questions which may
lead to unexpected outcomes. However, planning and evaluation are "musts"
if any success or measure of success is to be obtained.
Organizational and evaluative aids are included in each section of the
activity outlines. The introductions state general objectives. The ques-
tions provide direction (but not in a dictational manner) and suggest
behavioral changes which might be observed for evaluative purposes. Pro-
cedures, past examples, and limitations include guidance in organizing the
activity.
The foundation for planning and evaluation must begin at the time the
activity is selected. This is accomplished by identifying the desired out-
comes or objectives on paper in behavioral form. General statements such
as the following are of limited use:
a. The student will understand the effects of chemicals on a specific
ecosystem.
b. The students will develop an appreciation of the economic factors
associated with pollution.
However, behavioral statements which not only designate the desired
outcome but also identify a resulting behavior and the conditions under
which it is to be observed and evaluated can be extremely useful in planning
and evaluative efforts. For example:
When provided alkalinity, iron, and dissolved oxygen data for
a body of water, the student correctly describes one or more
biological effect which might be observed at the site.
These objectives should include not only cognitive or psychomotor cate-
gories but also affective behavior. While the former are significant, the
urgency of current pollution justifies emphasis of attitudes, values, and
motivation. Other than these remarks, it is not within the scope of this
guide to cover thoroughly the philosophy and writing of behavioral objectives.
Such information is readily available (see Bibliography) and should be con-
sulted by those who are unfamiliar with behavioral objectives.
As stated earlier, preliminary behavioral objectives should be formulated
immediately upon selection of the activity. In addition to serving an
evaluative role, they are a source of guidance for planning the unit. How-
ever, they must not be allowed to limit innovative approaches to the study.
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Appendix 4
Once the activity is in progress, the preliminary objectives should be
modified. If the activity is allowed to progress creatively, many unexpected,
desirable goals will emerge. They must not be ignored because they were
not recognized at the start. To the contrary, such goals should be incor-
porated into the list of objectives. It may be necessary to discard some
of the preliminary behavioral objectives if they are found inappropriate.
In this way the objectives evolve throughout the entire activity. As such
they become representative of the activity rather than a list of idealistic
goals developed for a file.
The revised behavioral objectives are used as guides in the formulation
of an evaluative tool appropriate to the study. If properly constructed,
the criteria and methods of evaluation will be stated within each objective.
In addition to fulfilling immediate planning and evaluative needs, the
resulting behavioral objectives and data provide valuable tools for
modifying the activity for future students.
References
Eiss, A., Behavioral Objectives in the Affective Domain, National Science
Teachers Association, Washington, D. C., 1969. This useful guide
for writing behavioral objectives includes specific examples encom-
passing psychometor, cognitive and affective domains.
Koran, J., et_ al_, How to Write Behavioral Objectives in Science
Instruction, National Science Teachers Association, Washington,
D. C., 1969. A discussion of the theory and expression of objectives
concerned with affective behavior is presented.
Mager, R. F., Preparing Instructional Objectives, Fearon Publishers,
Palo Alto, Calif., 1962.This is a programmed text for instruction
in the writing of objectives.
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Appendix 5
Bibliography
During this age of the "information explosion," it is imperative that
students learn to use many appropriate sources while researching a par-
ticular problem. For this reason a listing of those books and pamphlets
found particularly useful by teachers and students in the study of water
pollution during the 1969 and 1970 summer programs and the 1969-1970
school year are included. It is hoped that most of the publications
listed under Core References will be made available to students while
studying water pollution. The references listed under Additional Refer-
ences are also excellent, but due to prior consideration on the technical
level should probably be added only after the Core References have been
secured.
This bibliography is by no means comprehensive. Hopefully, current
periodicals, state and federal water surveys, for your area, and local
library resources will supplement this reference listing.
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Appendix 5
A. Core References
1. The following set of publications will provide a working reference
source for any class, upper elementary through 12th grade. It is
recommended that multiple copies of the asterisked publications be
obtained, depending on class size and grade level. Those refer-
ences with one asterisk (*) should be purchased in multiple copies
for elementary through 8th grade use. The double asterisk (**)
indicates that multiple copies could best be used for a 9th through
12th grade situation.
a. **American Public Health Association, Standard Methods for the
Examination of Water and Wastewater, (13th ed.), American
Public Health Association, Inc., 1740 Broadway, New York,
N. Y., 1971. This is an indispensible technical refer-
ence which includes test procedures and explanations of
dissolved solids and gases found in water. The biology
section includes collection methods and diagrams of
organisms.
b. Billings, W. D., Plants, Man, and the Ecosystem, (2nd ed.),
Fundamentals of Botany Series, Wadsworth Publishing Co.,
Belmont, California, 1970. 160 pp. This paperback can
introduce ecological concepts to 9th-12th graders and
contains a section concerning man's effects on the
environments.
c. Carvajal, J., and M. E. Munzer, Conservation Education—A
Selected Bibliography, The Interstate Printers and Pub-
lishers, Danville, 111., 1968. 98 pp. This is an
annotated bibliography dealing with not only water but
also air, population, and land conservation.
d. Frost, T. P., The Galloping Ghost of Eutrophy, Society for the
Protection of New Hampshire Forests, 5 South State St.,
Concord, N. H., 1968. 36 pp. This is a well written
pamphlet on the problem of eutrophication.
e. Leopold, L. B., and K. S. Davis, Water, Life Science Library,
Time Inc., New York City, 1966. 200 pp. This is a well
illustrated book covering water use and hydrology
(particularly useful for lower grade use).
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Appendix 5
f. **McKee, J. E., and H. W. Wolf, Water Quality Criteria,
(2nd ed.), Water Quality Control Board, Sacramento,
Calif., 1963. 548 pp. This is an excellent reference,
although dated, on chemical, biological, radioactive,
and pesticide pollutants. Each pollutant is explained
and many toxicity levels are listed. 3827 references
are listed.
g. **Heedham, J. G.,and P. R. Needham, A Guide to the Study of
Fresh-Water Biology, Holden-Day, Inc., 500 Sansome St.,
San Francisco7 Calif., 1969. 108 pp. This is an ex-
cellent reference book for any fresh-water biology work
which is thoroughly illustrated with a good section on
collection and equipment.
h. Ward, R. C., Principles of Hydrology, McGraw-Hill Book Co.,
New York City, 1967.Easily readable for high school
level, it contains stimulating ideas for activities.
i. Environmental Education for Everyone—Bibliography of Curricu-
lum Materials for Environmental Studies, National Science
Teachers Association, 1201 Sixteenth St., N. W.,
Washington, D.C., 20036, 1970. 36 pp. The most com-
prehensive bibliography reviewed to date, it includes
programs in environmental education curriculum guides,
textbooks, experiments, enrichment readings, periodical
listings, film strips, film lists, and other invaluable
materials for environmental studies.
j. Simplified Laboratory Procedures for Wastewater Examination,
WPCF Publication #18, Water Pollution Control Federation,
3900 Wisconsin Ave., Washington, D.C., 30016, 1969. 62
pp. Simplified Standard Method tests for physical and
chemical examination of water and is best for use in
grades 9-12.
2. The following publications may be purchased through the Superin-
tendent of Documents, U.S. Government Printing Office, Washington,
U. C. 20402.
a. Austin, John H., A Primer on Waste Water Treatment, 1969.
24 pp., $.55. This is a clearly written pamphlet con-
cerning the methods and problems of waste water
treatment which is good for 7th-12th grade use.
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Appendix 5
b. *Baldwin, H. L.,and C. L. McGuinnes, A Primer on Ground
Water. 1966. 26 pp., $.25. This pamphlet explains
ground water and ground-water resources competently
for 7th-12th grade use.
c. *Leopold, L. B.,and W. B. Langbein, A Primer on Water,
1966. 50 pp., $.35. This is a good introductory
pamphlet on hydrology and water use, is good for
7th-12th grade use, and includes a glossary.
d. Ingram, W. M., K. M. MacKenthum and A. F. Bartsch,
Biological Field Investigative Data for Uater
Pollution Surveys, 1966. 139 pp., $.70.This paper-
back has sections on graphical expression of data and
organism response to organic pollution and has both
good references and a glossary.
e. Thomas, H. E., The Yearbook of Agriculture, 1955: Water.
751 pp., $2.00. It contains numerous papers on water
source and use and is an excellent book for the money...
if it is still available from the Superintendent.
f. The Practice of Uater Pollution Biology, 1969. 281 pp.,
$1.50. This paperback covers areas such as aquatic
environments, organic wastes, toxic materials, acid
mine and radioactive wastes, eutrophication, marine
environments, water and waste treatment, and nuisance
organisms and has a large reference section.
g. Water Quality Criteria, Federal Water Pollution Control
Administration, 1968. 234 pp., $3.00. This book
defines the various uses of water in the U.S. and
recommends the various parameter limits for the
different water uses. Good references and glossaries
are included.
3. Individual copies of the following three pamphlets may be
obtained free from Mi Hi pore Corporation, Bedford, Massachusetts,
01730.
a. Experiments in Microbiology, 1969. 45 pp. An excellent
pamphlet for 7th-l2th grade use. Procedures and
experiments to familiarize students with microorganisms--
their occurrence in nature and ways to culture them--
are given.
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Appendix 5
b. Microbiological Analysis of Water, 1969. 25 pp. Good
procedures and explanations of coliform indicators
and other bacteria are discussed.
c. Microbiology for the Beginning Student, 1969. 12 pp.
Basic explanation of the membrane filtration technique
and equipment is presented.
4. The following three pamphlets may be purchased for $.50 each
through Educational Products Division, LaMotte Chemical Products
Company, Chestertown, Md., 21620.
a. *Amos, W. H., Limnology, 1969. 39 pp. This introduction
to the fresh water environment is simply written and
can be used by 7th-12th graders. Running and still
water and the organisms found in each are discussed.
b. Renn, C. E., A Study of Water Quality, 1968. 46 pp. This
is a good pamphlet dealing with the general aspects
of water pollution (usable in 7th-12th grade).
c. Renn, C. E., Our Environment Battles Water Pollution, 1969.
32 pp. This is a good paperback, usable in elementary
through 12th grade, dealing with various dissolved
solids and gases and their relationship with organisms.
B. Additional References
The following list of references could be extremely beneficial
in the study of water pollution. However, due to the price or
technical nature, these texts should probably be purchased after
the Core References have been obtained.
a. Chorley, R. J., (ed.), Water, Earth, and Man, Barnes and
Noble, Inc., New York City, 1969. This is a good
reference on hydrology and water use, capable of being
used by 7th-12th graders.
b. Foerster, J. W., "A Phyco-periphyton Collector," Turtox News,
47-3, pp 82-84, 1969. It describes a simple, easy-to-
make periphyton collector. The periphyton is collected
on glass microscope slides.
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Appendix 5
c. McHarg, I. L., Design With Nature, Natural History Press,
New York City, 1969. This is an excellent book which
deals with the need for environmental planning as
further development (of land) takes place (best for
high school use).
d. Pelczar, M. J.,and R. D. Reid, Microbiology, McGraw-Hill
Book Co., New York City, 19l>5~This is a microbiology
text which includes introductions to the taxonomy,
biochemistry, cultivation, control, and ecological
role of microorganisms.
e. Pennak, R. W., Fresh-Water Invertebrates of the United
States, Ronald Press Co., New York City, 1953.769
pp. This is a thorough, technical text.
f. Sawyer, C. N.,and P. L. McCarty, Chemistry for Sanitary
Engineers, (2nd ed.), McGraw-Hill Book CoV,"New York
City, 1968. 518 pp. A thorough presentation of the
theory and methods of sanitation chemistry is given.
This book might best be used as a teacher's reference
because a knowledge of elementary chemistry is assumed.
g. Smith, G. M., Fresh-Water Algae of the United States, McGraw-
Hill Book Co., New York City, 1950. 719 pp. This is
a comprehensive text on algae--collection, preservation,
and methods of study.
h. Ward, H. B.,and G. C. Whipple, Fresh-Water Biology, (2nd ed.),
John Wiley and Sons, New YorkTity, 1959. This is the single
most comprehensive manual for the identification of
aquatic plants and animals.
i. Special Publication Number 1, Sources of Limnological and
Qceanographic Apparatus and Supplies, American Society
of Limnology and Oceanography. Many specialized items
of biological collecting equipment are not available
from the usual supply houses. This publication lists
the suppliers. It is available from the Secretary of
the Society.
C. Periodicals
To keep students aware of environmental news, a number of
periodicals should be made available for classroom use. The
following periodicals were found useful during the 1969-70
school year:
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Appendix 5
a. The Conservation Foundation Letter, (monthly), The Con-
servation Letter, 1717 Massachusetts Ave., N. W.,
Washington, D. C. 20036, $6/yr. This is a three-
hole punched newsletter capable of being accumulated
for reference in a loose-leaf notebook and deals with
current legislation and all types of environmental
news. It is probably best for high school use.
b. Environmental Science and Technology, (monthly), American
Chemical Society, 20th and Northampton Sts., Easton,
Pa. 18042, $7/yr. This is a technical magazine but
is good for high school use.
c. Environment, (10 issues/yr.), Environment, Circulation
Dept., 438 North Skinker Boulevard, St. Louis, Mo.
63130, $8.50/yr. This is a well written periodical,
dealing in depth with current environmental problems
(capable of use in grades 7th-12th).
D. Movies
During the 1969-1970 school year, Miss Elizabeth Gage and her
students at Northfield School reviewed the following environ-
mental films. The film reviews worked on during the 1970
summer programs at Til ton have not yet been compiled but will
be forthcoming in the newsletters.
Title: It's Your Decision: Clean Water
Source: Association Films, Inc.
Regional Film Centers:
600 Grand Avenue, Ridgefield, N. J. 07657
561 Hillgrove Avenue, LaGrange, 111. 60525
324 Delaware Avenue, Allegheny County, Oakmont, Pa. 15139
Information: Color, 14 minutes, free loan.
Summary: The movie begins by briefly tracing the development of
towns and cities along rivers and streams. It shows how
rivers were able to handle their pollutional load, but
with our rapidly growing population, the pollution crisis
has become more acute. Animated figures are used to show
the basic characteristics of polluted water and how primary
and secondary water treatment plants operate.
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Appendix 5
Appraisal: Because the movie is easy to understand, it could be
shown to several different types of audiences. Clean
Water would be effective as an introductory film for a
pollution course. The film would be of benefit to
elementary school students if seen as an introduction
to our nation's pollution problems. The film would also
be very effective if shown in a community where a secondary
water treatment plant is needed. It could be shown to
the general public, too, possibly as a short feature in a
movie theatre.
Title: Oops, or How Broad Shoulders Polluted the River
Source: University of Minnesota, Audio-Visual Department, 2037
University Avenue, S. E., Minneapolis, Minn. 55455.
Information: Black and white (Color: $5.85), 22 minutes.
Summary: This movie gives insight into industry's problems in
controlling pollution, listing several different aspects
such as analyzing wastes, repairing equipment, showing
concern for waste treatment plants, and spreading re-
sponsibility to everyone. It stresses the importance on
competent, well-trained employees. The title refers to
a worker, "broad shoulders," who through inattentiveness,
poor management, and poor training created a big mess
of overflowing tanks of chemicals, petroleum, and suds.
Appraisal: The class felt that this movie was good for our purpose
but is best suited for its intended purpose of making in-
dustry aware of the problems of pollution and suggesting
how these problems can be solved. It was good for our
class because we have little opportunity to come into
contact with industry, their problems, and their attempts
to remedy the situations.
Title: Becket Adventures
Source: Sid Dupont, Becket Academy, East Haddam, Conn.
Information: Color, 30 minutes.
Summary: Mr. Dupont visits schools showing films or slides of the
canoe trip that Becket summer students take down the
Connecticut River from its source to the sound. The boys
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Appendix 5
on the trip surveyed points along the river, recording
animal and plant life .in the area, bottom structure,
flow, weather, geology and the condition of the river
itself. This information was written in a log which may
be obtained from Becket Academy.
Appraisal: Although this film was made by amateurs, we feel it
is useful for obtaining a general picture of the con-
dition of the Connecticut. We feel that Mr. Dupont
wants others to become aware of the environment in which
they live through this movie, just as the boys became
aware through their trip. We feel that this is a worth-
while movie for students of pollution to see.
Title: Story of a Lake
Source: Chevron Chemical Company, Advertising and Public Relations,
Ortho Division, 200 Bush Street, San Francisco, Calif.
Information: Color, 10-15 minutes, free loan (one day).
Summary: The film describes the conditions in a small lake choked
with water weed (Elodea) and includes interviews of home-
owners who describe the effect of the lake in this condi-
tion on their recreational enjoyment of the lake and on
property values of lake sites. The people try to control
the growth of water weed by raking it out but find that
this offers only temporary control. The lake community
calls on the services of a professional herbicide company
which sprays the lake with the chemical, Diquat, a product
of the Chevron Company. There are scenes showing the lake
being sprayed from a small boat. The effects of the chem-
.ical are described in rather general terms ("...attacks
green matter and breaks down cell structure..."). The
herbicide takes effect in 4-5 days and after 10-14 days
the water weed has decomposed. Views of the clean lake
are shown and community members are interviewed for their
impressions.
Appraisal: The film is clearly selling Diquat, a product of the
Chevron Chemical Company, and the biological content of
the film is minimal. It is, in our opinion, not as useful
for water pollution studies as it might be. One wonders
if the herbicide is "harmless to water animals," why the
men spraying the water are wearing coveralls and masks while
they are working. Further, we wonder specifically what
effect the chemical has on cells...maybe a letter to the
Chevron Company would clear this up for us.
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Appendix 5
Title: Membrane Microfiltration: A New Tool for Classroom Science
Source: Bernard I. Sohn, Educational Division, Millipore Corporation,
Bedford, Mass. 01730.
Also, Tilton School has this film to lend upon request.
Information: Color, 30 minutes, free loan.
Summary: The movie dealt specfically with Millipore apparatus, its
uses in the classroom, in the field, in industry, and in
medicine. It also stressed the advantage of the Millipore
over previous techniques of filtering by using clever pho-
tography. Animated figures representing bacteria described
the filtering process and its usefulness in detecting the
presence of micro-organisms.
Appraisal: Although this film was intended for showing to teachers,
we feel it should be used as an introduction for students
as well. The film is simple yet comprehensive in its
coverage of the subject.
Tit_1 e: Water: Pattern of Life
Source: Ohio Department of Natural Resources, Administrative
Services Section, 1500 Dublin Road, Columbus,
Ohio 43212.
Information: Color, free loan.
Summary: This movie told about the sources of, uses of, and problems
with natural water in the state of Ohio. It covered
many topics such as the hydrologic cycle, transportation,
droughts, flood control, underground water supply,
growing population, recreational uses of water, industrial
uses of water, reservoirs, and quality control of water.
It offered suggestions as how to solve problems; for example,
surveying, control of flood plains, and long range planning
of water use and supply. It also suggested that areas
should be divided into watershed basins to control the
problems.
Appraisal: This film would be good as an introduction to water
pollution or biology because it shows the wide usage of
water. It does not, however, deal specifically with pollu-
tion or control of it. It also is somewhat outdated because
it speaks of Lake Erie as a valuable source of water,
which it was at one time!!
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Appendix 5
Title: What Are We Doing to Our World (in two parts)
Source: University of Minnesota, Audio-Visual Department, 2037
University Avenue, S. E., Minneapolis, Minn. 55455.
Information: Color, 25 minutes each, $7.20 each reel.
Summary: "We are going to have to choose between the advancements
of technology. . ." and the necessity of maintaining eco-
logical balances on earth. This film explores some serious
consequences of unrestricted growth, pollution, waste, and
over-population problems.
What was a question of conservation on earth is becoming
a question of survival. Experiments such as the one at
Hubbard Brook attempt to understand ecological balances
and explore some of the unforeseen consequences of man's
attempts to make a better world.
Appraisal: This is an excellent film which provokes very good
discussion among students. It is an unbalanced film from
the point of view that it stresses what we are doing to
pollute and far less is said about what we might do to
prevent pollution. In several places, it is brought out
that people will have to re-examine their attitudes and
values concerning their relation to the earth environment
and this serves to balance the views presented. The
spokesman for the Agricultural Chemists presents some
interesting arguments about continued use of pesticides.
Title: Municipal Sewage Treatment Processes
Source: Communicable Diseases Center, Atlanta, Ga. 30333.
Information: Black and White, 13 minutes, free loan.
Summary: This movie begins with a definition of municipal sewage,
where it goes, and the results of its discharge. It goes
into great detail with "on location" scenes about the
different types of sewage treatment plants. The movie then
gives a basic outline of the steps a city or town must
follow in planning the design, construction, and main-
tenance of a plant. A simple basis for choosing the most
effective type of treatment is given and summarizes the
results that people can expect from their plant. The
movie also goes into some detail about the operation of
water treatment plants.
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Appendix 5
Appraisal: Since the movie is clear and easily understood in its
description of sewage and its treatment processes it could
be an effective way to inform the people of a town of the
need for a treatment plant and of the decisions they will
have to make. Though the movie is somewhat dated, it
could be used early in a water pollution course.
Title: The River Must Live
Source: Shell Film Library, 450 North Meridian Street,
Indianapolis, Ind. 46204.
Information: Color, 21 minutes, free loan.
Summary; This film deals with the growth of industry as a cause of
the growth in pollution. The fact that one person's
effluent is another's supply is mentioned. ' Processes of
decay of life in the river through bacteria and the1
different forms of life contained within an ecosystem are
discussed. The film speaks of how the pollutional imbalance
decreases the supply of oxygen. There is a need for time
in order to restore the natural balance. When there is too
much pollution the river dies. After being dumped in the
river, the pollution is carried to and lost in the sea's
vastness. From there, through the hydrocycle, the water
is purified and returned to the land. Different methods
of waste treatment can purify the water before it reaches
the sea. The film also treats the different uses of
water — industrial , commercial, and domestic.
Appraisal: This film was rated very good to excellent. The
narration was easy to follow, the organization was good,
and the music was appropriate. There was excellent visual
treatment of uses of water and different types of waste
water treatment. Also, the Shell Company refrained ad-
mirably from pushing its product. They did skim over the
possibility of oceanic pollution, ignoring the tertiary
treatment factor. The best use of this film would be as
an introduction in water pollution.
E. Equipment
Various companies are selling commercial water testing equipment
suitable for classroom use. Catalogs and descriptive literature
can be obtained from the following addresses.
1. Chemical testing equipment
a. Hach Chemical Company
Box 907
Ames, la. 50010
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Appendix 5
b. Delta Scientific Corporation
120 East Hoffman Ave.
Lindenhurst, N. Y. 11757
c. LaMotte Chemical Products Company
Educational Products Division
Chestertown, Md. 21620
2. Bacteriological equipment
Millipore Corporation
Educational Products Division
Bedford, Mass. 01730
3. Aquatic Biology Equipment
a. Oceanography Unlimited, Inc.
108 Main St.
Lodi, N. J. 07644
b. Wildlife Supply Company
2200 S. Hamilton St.
Saginaw, Mich. 48602
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Appendix 6
Water Pollution and Environmental Glossary
This glossary is a compilation of terms from aquatic ecological,
hydrologic and chemical fields of endeavor. Of the many persons
who contributed, the principle contributor was John E. Mathews
of the Department of the Interior (Robert S. Kerr Water Research
Center, Ada, Oklahoma).
Terms underscored in a definition are separately defined in this
Glossary. When appropriate, closely associated or related terms
are cited parenthetically, following the definition. Specific
synonyms are noted in parentheses with the listed word.
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Appendix 6
ABIOTIC FACTOR Physical, meteorological, geological, or chemical
aspect of environment.
ABYSSAL ZONE All of a sea or of a very deep lake below the bathyal
zone. The primary energy source for this region lies far above
in the euphotic zone; density of life depends on the amount of
organic material that settles from the euphotic zone. (See
Hadal zone.)
ACCLIMATION Physiological and behavioral adjustments of an organism
in response to a change in environment. (See Adaptation.)
ACCLIMATIZATION Acclimation of a particular species over several
generations in response to marked environmental changes.
ACID A hydrogen ion (H+) donor.
ACIDITY See Appendix 1: Chemistry.
ACTINOMYCETES Filamentous microorganisms intermediate between the
fungi and bacteria, although more closely related to the bacteria.
These organisms are widely distributed in soils and are often con-
spicuous in lake and river muds. They are often associated with
taste and odor problems in water supplies.
ACUTE TOXICITY Any toxic effect that is produced within a short period
of time, usually 24-96 hours. Although the effect most frequently
considered is mortality, the end result of acute toxicity is not
necessarily death. Any harmful biological effect may be the result.
(See Chronic Toxicity, Direct Toxicity.)
ADAPTATION Change in the structure, form or habits of an organism to
be better fit changed or existing environmental conditions. (See
Acclimation.)
AEROBIC Refers to life or processes occuring only in the presence of
free oxygen; refers to a condition characterized by an excess of
free oxygen in the aquatic environment. (See Anaerobic.)
ALGAE (Alga) Simple plants, many microscopic, containing chlorophyll.
Algae form the base of the food chain in aquatic environments.
Some species may create a nuisance when environmental conditions
are suitable for prolific growth.
ALKALINITY Appendix 1: Chemistry.
A-207
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Appendix 6
ALLOCHTHONOUS Pertaining to those substances, materials, or organisms
in a waterway which originate outside and are brought into the
waterway. (See Autochthonous.)
ALLUVIAL FAN (Delta)
ANABOLISM Synthesis or manufacture of organic compounds within an
organism. (See Metabolism.)
ANADROMOUS Pertaining to fishes that spend most of their life in salt
water but enter freshwater to spawn (e.g., salmon, shad, striped
bass, etc.). (See CatadromousT)
ANAEROBIC Refers to life or processes occurring in the absence of
free oxygen; refers to conditions characterized by the absence
of free oxygen. (See Aerobic.)
ANTAGONISM Reduction of the effect of one substance because of the
introduction or presence of another substance (e.g., one sub-
stance may hinder, or counteract, the toxic influence of another).
(See Synergism.)
APHOTIC ZONE That portion of a body of water to which light does not
penetrate with sufficient intensity to have any biological sig-
nificance. (See Euphotic Zone.)
ARTIFICIAL SUBSTRATE A device placed in the water (for a specified
period of time) that provides living spaces for a multiplicity
of organisms (e.g., glass slides, concrete blocks, multiplate
samplers, rock baskets, etc.). The primary purpose of arti-
ficial substrates is to allow the investigator to collect
organisms in areas where the physical habitat is limiting or
cannot be adequately sampled using conventional methods.
ASSIMILATION 1. Removal of dissolved or suspended materials from a
water mass by biological, chemical, and physical processes; 2.
Conversion or incorporation of absorbed nutrients into body
substances. (See Synthesis.)
ASSOCIATION All organisms occupying a given habitat.
ATOLL Large, thick, coral mass encircling a lagoon in tropical oceans;
sometimes portions of the reef become built up with sand, silt,
soil and vegetation to become an island. (See Barrier Reef,
Fringing Reef.)
AUFwUCHS (Periphyton)
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Appendix 6
AUTOCHTHONOUS Pertaining to those substances, materials, or organisms
originating within a particular waterway and remaining in that
waterway. (See Allochthonous.)
AUTOTROPHIC (Holophytic) Self nourishing; denoting those organisms
that do not require an external source of organic material but
can utilize light energy and manufacture their own food from
inorganic materials (e.g., green plants, pigmented flagellates).
(See Heterotrophic.)
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Appendix 6
BARRIER BEACH A ridge of deposits separated from the mainland by an
interval of water.
BARRIER REEF Large, thick, coral mass more or less surrounding an
island or paralleling the mainland shore in tropical areas and
separated from the land mass by a lagoon. (See Atoll, Fringing
Reef.)
BASE A hydrogen ion (H+) acceptor.
BATHYAL ZONE That region of the sea that extends from the euphotic
zone to the bottom of the continental slope. Density of life in
this zone depends on organic material settling from the euphotic
zone and is generally inversely proportional to the depth.
BEACH The zone of demarcation between land and water of lakes, seas,
etc., covered by sand, gravel or larger rock fragments.
BENTHIC REGION The bottom of a waterway; the substratum that supports
the benthos.
BENTHOS Bottom-dwelling organisms. These include: (1) sessile ani-
mals such as sponges, barnacles, mussels, oysters, worms, and
attached algae; (2) creeping forms such as snails, worms and in-
sects; (3) burrowing forms, which include clams, worms, and some
insects; and (4) fish whose habits are more closely associated
with the benthic region than other zones (e.g., flounders).
BIQASSAY A determination of the biological effect of some substance,
factor or condition employing living organisms or cells as the
indicator.
BIOCHEMICAL OXYGEN DEMAND See Appendix 1: Chemistry.
BIQCOENOSIS The plants and animals comprising a community.
BIOLOGICAL CONTROL 1. Use of natural predators, parasites, or patho-
gens to reduce or eliminate pest organisms (e-9-> use of gambusia
to feed on mosquito larvae). 2. Control of organisms by inter-
ference with their physiological processes (e.g., sterilization
of male flies).
BIOMASS The total amount of living material in a particular habitat
or area; an expression dealing with the total weight of a given
population of organisms.
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BIOMONITORING 1. Continuous surveillance of an effluent (or dilution
thereof) by using living organisms to test its suitability for dis-
charge into a receiving water. 2. Use of living organisms to
test the quality of a receiving water downstream from a waste dis-
charge. (See Bioassay.)
BIOTA All life of a region.
BIOTIC FACTORS (Biological Factors) In ecology, those environmental
factors which are the result of living organisms and their activities;
distinct from physical and chemical factors (e.g., competition, pre-
dation, etc.). (See Ecological Factor.)
BIQTIC POTENTIAL The inherent capability of an animal to multiply in
an unrestricted environment. (See Environmental Resistance.)
BIOTQPE (Habitat)
BLOODWORMS Midge fly larvae. Many of the species have hemoglobin
in the blood causing a red color and are often associated with rich
organic deposits. Also, the common name for certain of the marine
segmented worms (class Polychaeta). (See Sludgeworms.)
BLOOM A readily visible, concentrated growth or aggregation of minute
organisms, usually algae, in bodies of water.
BRACKISH WATERS Those areas where there is a mixture of fresh and salt
water; the salt content is greater than fresh water but less than
sea water; the salt content is greater than in sea water.
BUFFER SOLUTION A solution which, within limits, resists changes in pH.
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Appendix 6
CARNIVOROUS Pertaining to animals that feed on other animals. (See
Herbivorous.)
CARRYING CAPACITY The maximum quantity of organisms that any parti-
cular habitat can support over an extended period.
CATABOLISM The breakdown of organic compounds within an organism.
(See Metabolism.)
CATADROMOUS Pertaining to fish that spend most of their life in
freshwaters but migrate to the sea to spawn (e.g., american eel).
(See Anadromous.)
CATASTROPHIC DRIFT Massive drift of bottom organisms under conditions
of stress such as floods or toxicity. (See Drift Organisms,
Incidental Drift, Periodic Drift.)
CHEMICAL STRATIFICATION A layering of water in a lake because of
density differences owing to the varying or differential concen-
trations of dissolved substances with depth. (See Stratification.)
CHLOROPHYLL Green photosynthetic pigment present in many plant and
some bacterial cells. There are seven known types of chlorophyll;
their presence and abundance vary from one group of photosynthetic
organisms to another.
CHRONIC TOXICITY Toxicity, marked by a long duration, that produces an
adverse effect on organisms. The end result of chronic toxicity
can be death although the usual effects are sublethal (e.g.,
inhibits reproduction, reduces growth, etc.). These effects are
reflected by changes in the productivity and population structure
of the community. (See Acute Toxicity.)
CLASSIFICATION The placing of organisms into taxa (or categories)
according to established scientific requirements. (See Taxonomy,
Taxon.)
CLEAN WATER ASSOCATION An a^socja_vion_ of organisms found in any
natural, unpolluted environment. These associations are character-
ized by the presence of species that are sensitive to environmental
changes caused by introduction of pollutants. In many cases the
presence of a wide variety of species with relatively few individuals
representing any one of them is also characteristic. (See Sensitive
Organisms, Tolerant Association.)
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Appendix 6
CLINOMETER The standard instrument used by geographers to measure the
slope of a hillside.
COASTAL PLAIN A plain between the sea and higher land, usually at a
low elevation.
COASTAL WATERS Those waters surrounding the continent which exert a
measurable influence on uses of the land and on its ecology. The
Great Lakes and the waters to the edge of the continental shelf.
COASTAL ZONE Coastal waters and adjacent lands which exert a measur-
able influence on the uses of the sea and its ecology. The zone
extends onshore to the upper reaches of the tidal zone and adjacent
shore areas. (See Estuary.)
COLD-BLOODED ANIMALS Animals that lack an internal temperature re-
gulating mechanism to offset external temperature changes. Their
body temperature fluctuates to a large degree with that of their
environment. Examples are fish and aquatic invertebrates.
COLONY A distinguishable localized population within a species.
COMMUNITY All forms of life inhabiting a common environment.
COMPENSATION LEVEL The depth of a waterway at which there is a balance
between photosynthesis and respiration.
COMPETITION The effort of two or more individuals or species of a
community to utilize some of the same environmental resources.
COMPETITION EXCLUSION PRINCIPLE (Gause's Rule) No two species can
occupy the same niche at the same time.
CONSUMERS Organisms which feed upon other organisms; often divided
into first order consumers (Herbivores), second order (or higher)
consumers (Carnivores which eat primary consumers), etc. (See
Heterotrophic, TrophTc Level.)
CONTINENTAL SHELF The shallow, gently sloping portion of the sea
bottom bordering a continent, down to a depth of about 200
meters.
CONTINENTAL SLOPE The steeply sloping portion of the sea bottom ex-
tending seaward from the continental shelf.
CORAL A marine member of the phylum Coelenterata which secretes a
hard exoskeleton, chiefly of calcium carbonate.
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Appendix 6
CORAL REEF Large coral mass associated with coastal areas in the
tropics. (See Barrier Reef, Fringing Reef, Atol1.)
CRITERIA (Water Quality Criteria)
CRITICAL LEVEL (Threshold)
CRITICAL RANGE In bioassays, the value range of any factor between
the maximum level or concentration at which no organisms die to
the minimum level or concentration at which all organisms die
under a given set of conditions in a given period of time.
CULTURAL EUTRQPHICATION Acceleration by man of the natural process of
enrichment (aging7 of bodies of water.
CULTURE Cultivation of organisms in a medium containing necessary
nutrients.
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Appendix 6
D
DECOMPOSERS (Reducers)
DELTA (Alluvial Fan) A fan-shaped deposition of silt, sand, gravel
or other materials from a stream which occur when the hydraulic
gradient lessens abruptly, as in the discharge of a stream into
a lake or of a river into an ocean.
DENSITY (Population, Species) The number of individuals in relation
to the space in which they occur; refers to the closeness of
individuals to one another.
DENSITY STRATIFICATION (Stratification)
DEPOSITING SUBSTRATES Bottom areas where solids are being actively
deposited. These often occur in the vicinity of effluent dis-
charges. (See Sludge Deposits.)
DETRITUS Fragments of detached or broken down material.
DIFFUSION The even mixing of one compound throughout another.
DIRECT TOXICITY Toxicity that has an effect on organisms themselves
instead of having an effect by actual alteration of their habitat
or interference with their food supply. (See Acute Toxicity,
Chronic Toxicity, Indirect Toxicity.)
DISSOCIATION The separation of preexisting ions during the process
of solution.
DISSOLVED OXYGEN See Appendix 1 : Chemistry.
DISSOLVED SOLID Any substance which existed primarily as a solid
prior to the solution process.
DIURNAL 1. Refers to an event, process, or specific change that
occurs every day, usually associated with changes from day to
night. 2. Pertaining to those organisms that are active during
day time. (See Nocturnal.)
DIVERSITY Pertaining to the variety of species within a given
association of organisms. Areas of high diversity are character-
ized by a great variety of species; usually relatively few
individuals represent any one species. Areas with low diversity
are characterized by a few species; often relatively large numbers
of individuals represent each species.
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Appendix 6
DOMINANT Species which by their activity, behavior, or number, have
considerable influence or control upon the conditions of existence
of associated species; species which "controls" its habitat and
food web. (See Predominant.)
DRIFT ORGANISMS Benthic organisms temporarily suspended in the water
and carried downstream by the current. (See Inci dental Drift,
Periodic Drift, Catastrophic Drift.)
DYSTROPHIC LAKES Shallow lakes with brown water, high organic: matter
content, low nutrient availability, poor bottom fauna, and high
oxygen demand; oxygen is continually depleted and pH is usually
low. In lake aging the "age" between a eutrophic lake and a
swamp.
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Appendix 6
EBB TIDE That period of tide between a high water and the succeeding
low water; falling tide. (See Flood Tide.)
ECOLOGICAL FACTOR Any part or condition of the environment that in-
fluences the life of one or more organisms. (See Biot'ic Factor.)
ECOLOGICAL NICHE The role of an organism in the environment, its
activities and relationships to the living and nonliving environ-
ment; food and nutrition relationships are of primary importance.
(See Habitat Niche.)
ECOLOGY Interrelationships between organisms and their environment,
abiotic and biotic.
ECOSYSTEM A community, including all the component organisms, together
with the environment, forming an interacting system.
r£r.OTYPE (Habitat Form) The growth form or appearance of an organism
which is characteristic of a specific habitat. (Individuals of
the same species may appear different in various habitats.)
EMERSED (Emergent) AQUATIC PLANTS Plants that are rooted at the
bottom of a body of water, but project above the surface (e.g.,
cattails, bulrushes, etc.). (See Floating Aquatic Plants,
Submersed Aquatic Plants.)
END-POINT The point at which a titration is to be terminated, some-
times signifying the presence of equivalent amounts of reactants.
ENRICHMENT An increase in the quantity of nutrients available to
aquatic organisms for their growth and development. (See
Eutrophication.)
ENVIRONMENT All external influences and conditions affecting the life
and development of an organism.
ENVIRONMENTAL RESISTANCE Restriction imposed on the numerical in-
crease of a population by environmental factors. (See Biotic
Hutgntial.)
EPILIMNION The water mass extending from the surface to the ther-
mocline in a stratified body of water; the epilimnion is less
dense than the lower waters and is wind-circulated and essentially
homothermous. (See Hypo!imnion.)
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Appendix 6
EQUILIBRIUM 1. The condition in which a population or community is
maintained with only minor fluctuations in composition over an
extended period of time. Sometimes called Dynamic equilibrium.
2. A dynamic interaction of two opposing chemical or physical
processes occurring at equal rates.
ESTUARY That portion of a coastal stream influenced by the t_i_de_ of
the body of water into which it flows; a bay, at the mouth of a
river, where the tide meets the river current; an area where
fresh and marine waters mix. (See Positive Estuary, Inverse
Estuary, Neutral Estuary, Coastal Zone.)
EULITTQRAL ZONE (Tidal Zone)
EUPHOTIC ZONE The lighted region of a body of water that extends
vertically from the water surface to the depth at which
photosynthesis fails to occur because of insufficient light
penetration.
EURY- Prefix meaning wide (e.g., euryhaline refers to a wide range
of salinity tolerance; eurythermal refers to a wide range of
temperature tolerance). (See Steno-.)
EUTROPHIC LAKES Lakes which are rich in nutrients and organic materi-
als, therefore, highly productive. These lakes are often shallow
and seasonally deficient of oxygen in the hypo!imnion. (See
Oligotrophic Lakes.)
EUTROPHICATION The natural process of the maturing (aging) of a lake;
the process of enrichment with nutrients, especially nitrogen and
phosphorus, leading to increased production of organic matter.
(See Cultural Eutrophication, 01igotrophic Lakes, Eutrophic Lakes.)
EVAPOTRANSPIRATION The total of the transpiration of the plants of
an area plus the evaporation of water from the area equals the
total loss in the form of vapor.
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Appendix 6
FALCULTATIVE Refers to the capability of an organism to live under
varying conditions (e.g., a falcultative anaerobe is an organism
that although usually living in the presence of free oxygen can
live in the absence of free oxygen). (See Obligate.)
FALL OVERTURN A physical phenomenon that may take place in a body of
water during early autumn. The sequence of events leading to
fall overturn include: (1) cooling of surface waters, (2) density
change in surface waters producing convection currents from top to
bottom, (3) circulation of the total water volume by wind action,
and (4) vertical temperature equality. The overturn results in a
uniformity of the physical and chemical properties of the entire
water mass. (See Spring Overturn.)
FATHOM A unit of measurement equal to 6 feet (1.83 meters).
FAUNA Animal life.
FECAL COLIFORM See Appendix 1: . Bacteriology.
FECAL STREPTOCOCCUS See Appendix 1 : Bacteriology.
FIRTH A narrow arm of the sea; also the opening of a river into the
sea. (See Estuary.)
FJORD (Fiord) A narrow arm of the sea between highlands. (See Firth,
Estuary.)
FLOATING AQUATIC PLANTS Rooted plants that wholly or in part float
on the surface of "the water (e.g., water Tillies, water hyacinth
and duckweed). (See Emersed Aquatic Plants, Submersed Aquatic
Plants.)
FLOOD TIDE That period of tide between low water and the succeeding
high water; a rising tide. (See Ebb Tide.)
FLORA Plant life.
FOOD CHAIN Dependence of a series of organisms, one upon the other,
for food. The chain begins with plants and ends with the largest
carnivores (e.g., phytoplankton, zooplankton, forage fish, game
fish). Food chains usually do not exist in nature; they are parts
of food webs.
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Appendix 6
FOOD CYCLE (Food Web) All the interconnecting food chains in a
community.
FORAGE FISH Fish, usually smaller species, that are important as
food for other species.
FREE-SWIMMING (Motile) Actively moving about in water or capable of
moving about in water. (See Sessile.)
FRINGING REEF Large coral mass at the edge of any land mass in
tropical seas; it begins at the water's edge and may extend out
to a quarter mile. (See Barrier Reef, Atol1.)
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Appendix 6
GAME FISH (Sport Fish) Those species of fish considered to possess
sporting qualities on fishing tackle (e.g., salmon, trout, black
bass, striped bass, etc.). Game fish are usually more sensitive
to environmental changes than rough J'ish.
CAUSE'S RULE (Competition-Exclusion Principle)
GROUND WATER The body of water derived from percolation; retained
in the soil, bub-soil and underlying rocks of an area.
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Appendix 6
HABITAT (Biotype) A specific type of place that is occupied by an
organism, a population, or a community.
HABITAT FORM (Ecotype)
HABITAT NICHE The specific part or smallest unit of a habitat occu-
pied by an organism. (See Ecological Niche.)
HADAL ZONE Pertaining to that part of the ocean at depths exceeding
6,000 meters, including both water and floor or bottom. (See
Abyssal Zone.)
HERBIVORE An organism that feeds on plant material; a first order
consumer.(See Carnivore.)
HETEROGENEOUS Consisting of dissimilar elements or constituents.
(See Homogeneous.)
HETEROTRQPHIC (Holozoic) Pertaining to organisms that are dependent
on organic material for food. (See Autotrophic.)
HIGHER AQUATIC PLANTS (Pond Weeds) Those plants whose seeds germinate
in the water phase or substrate of a body of water and which must
spend part of their life cycle in water. This grouping includes
plants which grow completely submersed as well as a variety of
emersed and floating leaf types.
HQLOPHYTIC (Autotrophic)
HOLQZOIC (Heterotrophic)
HOMOGENEOUS Of uniform composition throughout.
HOMQTHERMOUS Having the same temperature throughout.
HYDROLYSIS The reaction of a salt with water to produce a basic or
acidic solution.
HYPOLIMNION The region of a body of water that extends from the
thermocline to the bottom and is essentially removed from major
surface influences. (See Epilimnipn.)
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Appendix 6
I
IDENTIFICATION The use of a taxonomic key or the equivalent to de-
termine the scientific name of an organism.
INCIDENTAL DRIFT The casual, random drift of organisms. (See Drift
Organisms, Catastrophic Drift, Periodic Drift.)
INDICATOR 1. A substance which, by means of a color change, identi-
fies the end-point of a titration. 2. A substance which, by
means of a color change, qualitatively and/or quantitatively
evaluates the presence of an unknown substance.
INDIRECT TOXICITY Toxicity that affects organisms by interfering
with their food supply or modifying their habitat instead of
directly acting on the organisms themselves"! (See Direct
Toxicity.)
INFILTRATION The term used by hydrologists to describe the gradual
downward flow of water from the surface through soil to ground
water and water table reservoirs.
INLET A short, narrow waterway connecting a bay, lagoon, or similar
body of water with a large parent body of water; a stream which
flows into a lake; an arm of the sea, or other body of water,
that is long compared to its width and that may extend a con-
siderable distance inland.
INSTAR^ A stage in the life cycle of an insect or other arthropod
between two successive molts.
INTERACTION Mutual or reciprocal action or influence between organisms,
between organisms and environment, or between environmental
factors.
INTERSPECIFIC Refers to relations or conditions between species.
(See Intraspecific.)
INTERTIDAL ZONE (Tidal Zone)
INTOLERANT ORGANISMS (Sensitive Organisms)
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Appendix 6
INTRASPECIFIC Refers to relations or conditions between individuals
within a species. (See Interspecific.)
INVERSE ESTUARY Type of estuary in which evaporation exceeds the
supply of freshwater; evaporation freshwater inflow + pre-
cipitation. (See Positive Estuary, Neutral Estuary.)
INVERTEBRATES. Animals without an internal skeletal structure (e.g.,
insects, mollusks, crayfish). (See Vertebrate.)
K)N An atom or group of atoms which has become charged either by
loss or by gain of one or more electrons.
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Appendix 6
LAGOON 1. A shallow sound, pond, or channel near or communicating
with a larger body of water. 2. A settling pond for treatment
of wastewater.
LARVA The immature form of an animal which is unlike its parents.
Larvae are usually self-feeding but must pass through some sort
of metamorphosis before assuming the characteristics of the
adult; in insects, the wormlike stage between the egg and the
pupa.
LAW OF THE MINIMUM. LIEBIG'S "The growth and reproduction of an
organism is dependent on the nutrient substance, such as oxygen,
carbon dioxide, calcium, etc., that is available in minimum
quantity." (See Limiting Factor.)
LAW OF TOLERANCE, SHELFORD'S "When one environmental factor or con-
dition is near the limits of toleration, either minimum or
maximum, that one factor or condition will be the controlling
one and will determine whether or not a species will be able to
maintain itself." (See Limiting Factor.)
LEACHING The process by which nutrients in the soil are dissolved
and carried away by water flowing through it by processes such
as percolation.
LENTIC Pertaining to standing (nonflowing) waters such as lakes,
ponds, and swamps. (See Lotic.)
LIFE CYCLE The various phases, changes, or stages through which an
individual passes from the fertilized egg to death of the mature
organism. Briefly stated it is birth-maturation-reproduction-
death.
LIMITING FACTOR A factor whose absence, or excessive concentration,
exerts some restraining influence upon a population through
incompatibility with species requirements or tolerance. (See
Law of the Minimum, Law of Tolerance.)
LIMNETIC ZONE The open-water region of a lake, especially in areas
too deep to support rooted aquatic plants. This region supports
plankton and fish as the principal plants and animals. (See
Littoral Zone.)
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Appendix 6
LIMNOLOGY The ecology of fresh waters.
LITTORAL ZONE The shallow area that extends from shore to the lake-
ward limit of rooted aquatic plants; the shoreward region of a
body of water; in marine ecology, the tidal zone. (See Limnetic
Zone.)
LOT1C Pertaining to flowing waters such as streams and rivers.
(See Lentic.)
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Appendix 6
M
MACROQRGANISMS Those organisms retained on a U.S. standard sieve
No. 30 (openings of 0.589 mm); those organisms visible to the
unaided eye. (See Microorganisms.)
MACROPHYTE Any plant that can be seen with the naked, unaided eye
[e.g., aquatic mosses, ferns, liverworts, rooted plants, etc.).
MEDIAN TOLERANCE LIMIT (Tin,) The concentration of tested substance
in water at which just 50% of the test organisms survive for
a specified period of exposure. (See Tolerance Limit.)
MEROMICTIC LAKES Lakes in which dissolved substances create a grad-
ient of density differences with depth; this prevents complete
mixing or circulation of water masses. (See Chjemi caj_
Stratification.)
MEROMIXIS A condition of permanent stratification of water masses
in lakes.
MESOLIMNION (Thermocline)
METABOLISM The sum of all chemical processes occuring within an
organism; includes both synthesis (anabolism) and breakdown
(catabolism) of organic compounds.
METALIMNION (ThermoclIne)
METAMORPHOSIS Distinct transformation of an animal from one dis-
tinctive life history stage to another in its postembryonic
development (e.g., larva of an insect to a pupa). (See Life
Cycle.)
MICROORGANISMS Those organisms retained on a U.S. standard sieve No.
100 (openings of 0.149mm); those minute organisms invisible or
only barely visible to the unaided eye. (See Macroorganisms.)
MOLARITY A concentration unit which denotes the number of moles of
particles (molecules or ions) present in 1 liter of solution.
MOLE A collective unit which signifies 6 x 10" of anything but is
used primarily when dealing with molecules or ions.
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Appendix 6
MOLT To cast or shed periodically the outer body covering which
permits an increase in size. This is especially characteristic
of invejrtebrates. (See Ins tar.)
HOTIIE (Free swimming)
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Appendix 6
N
NANOPLANKTON Very minute plankton not retained in a plankton net
equipped with No. 25 silk bolting cloth (mesh, 0.03 to 0.04 mm).
NATURAL SELECTION Processes occurring in nature which result in
selective survival and elimination of individuals less well
adapted to their environment.
NAUPLIUS Free-swimming microscopic larval stage characteristic of
many crustaceans, barnacles, etc.
NEAP TIDES Exceptionally low tides which occur twice each month
when the earth, sun and moon are at right angles to each other-,
these usually occur during the moon's first and third quarters.
(See Spring Tides.)
NEKTON Macroscopic organisms swimming actively in water (e.g.,
fish). (See Plankton.)
NERITIC ZONE Relatively shallow water zone which extends from the
high-tide mark to the edge of the continental shelf.
NET PLANKTON Plankton retained in a plankton net equipped with No.
25 silk bolting cloth (mesh, 0.03 to 0.04 mm).
NEUSTON Organisms associated with, or dependent upon, the surface
film (air-water interface) of bodies of water.
NEUTRAL ESTUARY Type of estuary in which neither the freshwater
inflow nor the evaporation predominates; freshwater inflow +
precipitation = evaporation. (See Positive Estuary, Inverse
Estuary.)
NEUTRALIZATION The process of nullifying the effects of an acid
(base) through the addition of a base (acid), usually accompanied
by the formation of salt, water, and heat.
NICHE See Ecological Niche, Habitat Niche.
NOCTURNAL Pertaining to those organisms that are active at night.
(See Diurnal.)
NUISANCE ORGANISMS (Pests) Those organisms capable of interfering
with the use or treatment of water.
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Appendix 6
NUTRIENTS 1. Elements, or compounds, essential as raw materials
for organism growth and development (e.g., carbon, oxygen,
nitrogen, phosphorus, etc.). 2. The dissolved solids and
gasses of the water of an area.
NYMPH An immature developmental form characteristic of the pre-
adult stage in insects that do not have a pupal stage (e.g.,
May flies and stone flies). (See Larva.)
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Appendix 6
0
OBLIGATE Limited to one mode of life or action. (See Facultative.)
OCEANIC ZONE The region of open ocean beyond the continental shelf.
OLIGOTROPHIC LAKES Deep lakes which have a low supply of nutrients,
thus they support very little organic production. Dissolved
oxygen is present at or near saturation throughout the lake
during all seasons of the year. (See Eutrophic Lakes.)
OMBROTROPHY Air induced changes in water quality.
OMNIVOROUS Animal which is a first order consumer at some times and
a second or higher order consumer at others. (See Herbivorous,
Carnivorous.)
OPTIMUM LEVEL The most suitable degree of an environmental factor
for the full development of the organism concerned. (See
Tolerance Range.)
ORGANISM Any living individual.
OSMOREGULAnON The adjustment in the osmotic concentration of
solutes in body fluids in organisms to environmental conditions
(e.g., when salmon migrate from salt to freshwater).
OVERTURN The period of mixing (turnover) by top to bottom circulation
of previously stratified water masses. This phenomenon may occur
in spring and/or fall; the result is a uniformity of physical and
chemical properties of the water at all depths. (See Thermal
Stratification, Chemical Stratification, Spring Overturn, Fall
Overturn.)
OXIDATION The loss of electrons by an atom or ion_.
OXIDIZING AGENT A substance capable of accepting electrons from
another substance and, thereby, being reduced.
OXYGEN DEBT A temporary phenomenon that occurs in an organism when
available oxygen is inadequate to supply the respiratory demand.
During such a period the metabolic processes result in the accum-
ulation of breakdown products that are not oxidized until sufficient
oxygen becomes available.
OXYGEN_DEFICIT The difference between observed oxygen concentration
and the amount that would theoretically be present at 100% satu-
ration for existing conditions of temperature and pressure.
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Appendix 6
PARASITE An organism that lives on or in a host organism during all
or part of its existence. Nourishment is obtained at the expense
of the host.
PATHOGEN An organism or virus that causes a disease.
PELAGIC ZONE The open sea, away from the shore. Comparable with the
limnetic zone of lakes.
PERCOLATION Infiltration.
PERIODIC DRIFT Drift of bottom organisms at regular or predictable
intervals such as diurnal, seasonal, etc. (See Drift Organisms,
Catastrophic Drift. Incidental Drift.)
PERIPHYTON (Aufwuchs) Attached microscopic organisms growing on the
bottom, or other submersed substrates, in a waterway.
PESTICIDE Any chemical preparation used to kill pests. Include
insecticides, herbicides, fungicides, etc.
PESTS (Nuisance Organisms)
JDH_ See Appendix 1: Chemistry.
PHOTOSYNTHESIS The metabolic process by which simple sugars are
manufactured from carbon dioxide and water by plant cells using
light as an energy source. (See Chlorophyll.)
PHOTIC ZONE (Euphotic Zone)
PHYTOPLANKTON The plants of the plankton. Unattached microscopic
plants subject to movement by wave or current action. (See
Zooplankton.)
PLANKTON Suspended microorganisms that have relatively low powers
of locomotion or that drift in the water subject to the action
of waves and currents. (See Benthos, Periphyton, Nekton.)
POND WEEDS (Higher Aquatic Plants)
POOLS Areas of a stream, where the velocity of current is reduced.
The reduced velocity provides a favorable habitat for pjankton.
Silts end other loose materials that settle to the bottorrfof
pools are favorable for burrowing forms of benthos. (See Riffle.)
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Appendix 6
POPULATION A group of interacting individuals of the same species,
area, or community.
POSITIVE ESTUARY Coastal indentures in which there is a measurable
dilution of sea water by land drainage; freshwater inflow + pre-
cipitation evaporation. (See Inverse Estuary, Neutral Estuary.)
POTAMON ZONE Stream reach at lower elevations characterized by re-
duced flow, higher temperature, and lower dissolved oxygen levels.
(See Rithron Zone.)
ppm (parts per million) A unit of concentration equivalent to the
number of mi 11igrams of solute in 1 liter of solution.
PRECIPITATE 1. (noun) A solid which separates from a solution because
of some chemical or physical change. 2. (verb) The formation of
such a solid.
PREDATOR An animal that kills and consumes other animals. (See Prey.)
PREDOMINANT Those organisms that are of outstanding abundance in a
particular community for a given period of time. (See Dominant.)
PREY An animal that is killed and consumed by another animal. (See
Predator.)
PRIMARY PRODUCTIVITY The total quantity of protoplasm produced by
autotrophic organisms per unit of time in a specified habitat.
PRODUCERS Organisms that synthesize organic Material from inorganic
substances (e.g., plants). (See Consumers, Reducers.)
PRODUCTION The process of producing organic material; the quantity
produced.
PRODUCTIVITY Rate of protoplasm formation or energy utilization by
one or more organisms; total quantity of organic material produced
within a given period in a specified habitat.
PROFUNDAL ZONE The deep, Bottom-water area beyond the depth of
effective light penetration. All of the lake floor beneath the
hypgjiinnion.
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Appendix 6
PROLIFIC Pertaining to organisms that have a high reproductive
potential and normally produce large numbers of young.
PROTOPLASM The living material in cells of plants and animals.
PUPA An intermediate, usually quiescent, form following the larval
stage in insects, and maintained until metamorphosis to the
adult stage. (See Larva.)
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Appendix 6
QUALITY A term to describe the composite chemical, physical, and
biological characteristics of a water with respect to its
suitability for a particular use.
QUIESCENT Refers to the temporary cessation of development, move-
ment or other activity. (See Pupa.)
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Appendix 6
R
RAPIDS Areas of a stream where velocity of current is great enough
to keep the bottom clear of all loose materials, thus providing
a firm substrate. The surface of the water is disrupted by
turbulent currents. This area is occupied largely by specialized
benthic or periphytic organisms that can firmly attach or cling
to a firm substrate. (See Pools, Riffles.)
RED TIDE A visible red-to-orange coloration of an area of the sea
caused by the presence of a bloom of certain plankton. These
blooms are often the cause of major fish kills.
REDD A type of fish spawning area associated with flowing water and
clean gravel. Fishes that utilize this type of spawning area
include trout, salmon, some minnows, etc.
REDUCERS (Decomposers) Those organisms, usually bacteria or fungi,
that break down complex organic material into simpler compounds.
(See Producers, Consumers.)
REDUCING AGENT A substance capable of releasing electrons to another
substance, thereby, being oxidized.
REDUCTION The gain of .electrons by an atom or an ion.
REEF A ridge of rocks, sand, soil, or coral projecting from the
bottom to or near the surface of the water.
RESPIRATION The complex series of chemical and physical reactions in
all living organisms by which the energy and nutrients in foods
are made available for use. Oxygen is used and carbon dioxide
released during this process. (See Metabolism.)
RIFFLES Fast sections of a stream where shallow water races over
stones and gravel. Riffles usually support a wider variety of
bottom organisms than other stream sections. Also called rifts.
(See Pools. Rapids.)
RITHRON ZONE Stream reach at higher elevations characterized by rapid
flow, low temperature, and high dissolved oxygen levels. (See
Potamon Zone.)
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Appendix 6
ROUGH FISH Those species of fish considered to be of either poor
fighting quality when taken on tackle, or of poor eating quality
(e.g., carp, gar, suckers, etc.). These fish are considered un-
desirable in most situations. Most species in the group are
more tolerant of widely changing environmental conditions than
game fish.
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Appendix 6
SALT MARSH Low area adjacent to the sea that is covered with salt
tolerant vegetation and regularly flooded by the high tide;, similar
inland areas near saline springs or lakes, though not regularly
flooded.
SAPROB 1C Living on dead or decaying organic matter. (See Scavenger^)
SAPROBICITY The sum of all metabolic processes which are the direct
opposite of primary production; can be measured either by the
dynamics of metabolism or analysis of community structure.
SAPROBIENSYSTEM European system of classifying organisms according to
their response to organic pollution in slow moving streams: 1.
Alpha-Mesosaprobic Zone - Area of active decomposition, partly
aerobic, partly anaerobic, in a stream heavily polluted with organic
wastes; 2. Beta-Mesosaprobic Zone - That reach of a stream that
is moderately polluted with organic wastes; 3. Oligosaprobic Zone -
That reach of a stream that is slightly polluted with organTc
wastes and contains the mineralized products of self-purification
from organic pollution; but with none of the organic pollution
remaining; 4. Polysaprpbic Zone - That area of a grossly polluted
stream which contains the complex organic wastes that are de-
composing primarily by anaerobic processes.
SCAVENGER An organism that consumes decomposing organic matter.
SECONDARY PRODUCTIVITY Total quantity of animal (and other Hetero-
trophic) protoplasm produced per unit of time in a specified
habitat. (See Primary Productivity, Productivity.)
SEDIMENT The material that settles to the bottom of a waterway.
SEEPAGE Any flow of ground water to the surface of the land. This
can be in wells, springs, streams or in trickles of water we see
in areas such as roadside cuts.
SEICHE Periodic oscillations in the water level of a lake or inland
sea. These oscillations take place when a temporary local de-
pression or elevation of the water level occurs.
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Appendix 6
SENSITIVE ORGANISMS (Intolerant Organisms) Organisms that exhibit a
rapid response to environmental changes and are killed, driven out
of the area, or as a group are substantially reduced in numbers
when their environment is fouled. (See Tolerant Association.)
SESSILE Pertaining to those organisms that are attached to a sub-
strate and not free to move about (e.g., periphyton). (See Free-
swi turning.)
SESTON All material, both organic and inorganic, suspended in a
waterway.
SLOPE The term used to describe the steepness of a hillside. It is
often expressed in degrees (of an angle) or in per cent. A ten
per cent slope means an increase in altitude of 10 feet for every
100 horizontal feet traveled.
SLUDGE DEPOSITS Accumulations of settled, usually rapidly decomposing,
organic material in the aquatic system.
SLUDGEWORMS Aquatic segmented worms (class Oligochaeta) that exhibit
marked population increases in waters polluted with decomposable
organic wastes. (See Bloodworms.)
SPAWN 1. In aquatic animals, to produce or deposit eggs or sperm.
2. To produce eggs or young. 3. Eggs of fishes and higher
aquatic invertebrates.
SPECIES (Both singular and plural) An organism or organisms forming a
natural population, or groups of populations, that transmit specific
characteristics from parent to offspring. Each species is repro-
ductively isolated from other populations with which they might
breed. Hybrids, the results of interbreeding, usually exhibit a
loss of fertility.
SPORT FISH (Game Fish)
SPRING OVERTURN A physical phenomenon that may take place in a body
of water during the early spring. The sequence of events leading
to spring overturn include: (1) melting of ice cover, (2)
warming of surface waters, (3) density changes in surface waters
producing convection currents from top to bottom, (4) circulation
of the total water volume by wind action, and (5) vertical temper-
ature equality. The overturn results in a uniformity of the
physical and chemical properties of the entire water mass. (See
Fall Overturn, Overturn.)
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Appendix 6
SPRING TIDE Exceptionally high tide which occurs twice per lunar
month when there is a new or full moon, and the earth, sun,
and moon are in a straight line. (See Neap Tides.)
STANDARD (water Quality Standard)
STANDING CROP The quantity of living organisms present in an environ-
ment at a selected point in time.
STENO- Prefix denoting a narrow range of tolerance of an organism to
a specific environmental factor (e.g., stenothermal refers to
temperature; stenohaline refers to salinity; etc.). (See Eury-.)
STIMULUS An influence that causes a response in an organism. (See
Taxi s.)
STRATIFICATION (Density Stratification) Arrangement of water masses
into separate, distinct, horizontal layers as a result of differ-
ences in density; may be caused by differences in temperature,
dissolved or suspended solids. (See Thermal Stratification,
Chemical Stratification.)
SUBLITTORAL ZONE The part of the shore from the lowest water level
to the lower boundary of plant growth; transition zone from the
1ittoral to profundal bottom.
SUBMERSED (Submerged Aquatic Plants) Higher aquatic plants that
grow beneath the surface of the water (e.g., pondweed, coon-
tails, etc.).
SUBSTRATE The bottom material of a waterway; the base or substance
upon which an organism is growing; a substance undergoing oxidation.
SUMMER KILL Complete or partial kill of a fish population in ponds or
lakes during the warm months, variously produced by excessively
warm water, by a depletion of dissolved oxygen, and by the release
of toxic substances from a decaying algal bloom, or by a combination
of these factors. (See Winter Kill.)
SUPERSATURATION A condition in which a solution has more solute dis-
solved than is normally possible under the existing conditions.
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Appendix 6
SUPRALITTORAL ZONE (Supratidal Zone) The portion of the seashore
adjacent to the tidal or spray zone.
SURFACE AQUATIC PLANTS (Floating Aquatic Plants)
SUSPENDED SOLID Any solid substance present in water in an undis-
solved state, usually contributing directly to turbidity.
SYMBIOSIS Two organisms of different species living in close
association, one or both of which may benefit and neither is
harmed.
SYNERGISM The joint action of two or more substances is greater than
tfie sum of the action of each of the individual substances (e.g.,
action of certain combinations of toxicants). (See Antagonism.)
SYNTHESIS The production of a substance by the union of elements or
simpler chemical compounds.
SYSTEMATICS (Taxonomy)
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Appendix 6
TARN Small mountain lake or pond.
TAXIS Directed movement by an organism in response to a stimulus
(e.g., phototaxis is directed movement in response to a light
stimulus; thermotaxis is directed movement in response to heat
or cold as a stimulus; etc.).
TAXQN (Taxa) Any taxonomic unit or category of organisms (e.g.,
species, genus, family, order, etc.).
TAXONOMY (Systernatics) Organism classification with reference to its
relationship in the plant, animal, or protist kingdoms; includes
the bases, principles, procedures and rules of classification.
TERRITORY The area which an animal defends against intruders.
THERMAL STRATIFICATION The layering of water masses owing to differ-
ent densities in response to temperature. The condition of a
body of water in which the successive horizontal layers have
different temperatures, each layer more or less sharply differ-
entiated from the adjacent ones, the warmest (or the coldest) at
the top. (See Overturn.)
THERMOCLINE (Mesolimnion, Metalimnion) The transition zone between
the warm epilimnion and cold hypolimnion of stratified bodies of
water; temperature change equals or exceeds 1°C for each meter
of depth. (See Thermal Stratification.)
THRESHOLD (Critical Level) The maximum or minimum duration or in-
tensity of a stimulus that is required to produce a response in
an organism.
TIDAL FLAT The sea bottom, usually wide, flat, muddy and nonpro-
ductive, which is exposed at low tide.
TIDAL MARSH A low, flat marshland that is intersected by channels
and tidal sloughs, usually covered by high tides; vegetation
consists of rushes, grasses, and other salt tolerant plants.
(See Salt Marsh.)
TIDAL ZONE (Eulittoral Zone. Intertidal Zone) The area of a shore
between the limits of water level fluctuation; the area between
the levels of high and low tides.
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Appendix 6
TIDE The alternate rising and falling of water levels, twice in
each lunar day, due to gravitational attraction of the moon and
sun in conjunction with the earth's rotational force.
TITRATION The determination of the volume of a solution needed to
react with a known volume of sample, usually involving the
progressive addition of the solution to the sample until the
sample has reacted fully.
TLm (Median Tolerance Limit)
TOLERANCE Relative capability of an organism to endure an unfavorable
environmental factor.
TOLERANCE LIMIT (TL1Q...100) The concentration of a substance which
some specified portion of an experimental population can endure
for a specified period of time with reference to a specified type
of response (e.g., TL]QQ means that all test organisms endured
the stress for the specified time; TL-.Q means only 10% of the
test organisms could tolerate the imposed stress for the specified
time). (See Median Tolerance Limit.)
TOLERANCE RANGE The range of one or more environmental conditions
within which an organism can function; range between the highest
and lowest value of a particular environmental factor in which
an organism can live.
TOLERANT ASSOCIATION An association of organisms capable of with-
standing adverse conditions within the habitat. This association
is often characterized by a reduction in the number of species
(from a clean water association) and, in the case of organic
pollution, an increase in individuals representing certain
species.
TOXICANT A substance that through its chemical or physical action,
kills, injures, or impairs an organism; any environmental factor
which, when altered, produces a harmful biological effect.
(See Pesticide.)
TOXICITY Quality, state, or degree of the harmful effect resulting
from alteration of an environment factor.
TOTAL COL I FORM See Appendix 1: Bacteriology.
TRIPTON The dead suspended particulate matter in aquatic habitats;
the nonliving portion of the seston. (See Detritus.)
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Appendix 6
TROPHIC LEVEL One of the parts in a nutritive series in an ecosystem
in which a group of organisms in a certain stage in the food
chain secures food in the same general manner. The first or
lowest trophic level consists of producers (green plants); the
second level of herbivores; the third level of secondary
carnivores. Most bacteria and fungi are organisms in the
reducer (decomposer) trophic level.
TROPHOGENIC REGION The area of a body of water where organic
production from mineral substances takes place on the basis
of light energy and photosynthetic activity.
TRANSPIRATION The photosynthetic and physiological process by which
plants release water into the air in the form of water vapor.
TURBIDITY See Appendix 1: Chemistry.
TURNOVER (Overturn)
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Appendix 6
U
UBIQUITOUS ORGANISMS Organisms that can tolerate a wide range of
environmental conditions or variation; organisms that are so
active or numerous as to seem to be present or existent in all
types of environments. (See Tolerant Association, Sensitive
Organisms.)
UNICELLULAR Refers to an organism that consists of only one cell
(e.g., blue-green algae, protozoa, bacteria). These organisms
may, however, be filamentous or colonial in form.
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Appendix 6
VERTEBRATES Animals that have an internal skeletal system. (See
Invertebrate.)
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Appendix 6
W
WATER QUALITY CRITERIA "A scientific requirement on which a decision
or judgement may be based concerning the suitability of water
quality to support a designated use." (See Water Quality
Standard.)
WATER QUALITY STANDARD "A plan that is established by governmental
authority as a program for water pollution prevention and
abatement." (See Water Quality Criteria,.)
WATERSHED The area of land delineated by the line separating areas
of land, the water from which drains into separate river or
stream systems.
UATER TABLE The level of ground water of an area.
WINTER KILL The death of fishes in a body of water during a prolonged
period of ice and snow cover; caused by oxygen exhaustion due to
respiration and lack of photosynthesis. (See Summer Ki11.)
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Appendix 6
ZONE An area characterized by similar flora or fauna; a belt or
area to which certain species are limited.
ZOOPLANKTON The animals of the plankton. Unattached microscopic
animals having minimal capability for locomotion.
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Appendix 7
Laboratory and/or Field Safety
The items below are arranged according to areas of application;
the general comments apply to most areas.
A. General Comments
1. When heating liquids or when working with acids, bases, or
other caustic liquids, wear goggles or some other form of
eye protection.
2. Show concern for others by not pointing the opening of a
container (which you are heating or to which you are adding
chemicals) in the direction of fellow workers or their work.
3. Looking directly into containers which are being heated is
very dangerous.
4. When such things as strong acids and strong bases are mixed
with water or with each other, large amounts of heat are
generated. Therefore, use pyrex containers and do not hold
them in your hands. Leaving them unattended while they are
still hot may cause injury to others.
5. Always pour acid into water, never water into acid.
6. If spillage occurs, turn off open flames and hot plates.
Clean up spills immediately. The following procedure should
be followed for cleaning up acids, bases, and other caustic
substances.
a. Get paper towels and a large beaker partially filled
with water.
b. Grasp two or three folded paper towels from one end;
daub and swab with the other end.
c. Place wet towels in the beaker.
d. When the liquid is cleaned up, wipe the area with a
damp sponge or several thicknesses of wet paper towels.
e. Take beaker, wet paper towels, and sponge to sink and
rinse with lots of water.
f. Squeeze wet paper towels and place in waste basket.
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Appendix 7
7. Acids, bases, caustic substances, and water samples which
might be contaminated should not be pipetted by drawing
the liquid into the pipette with your mouth. If a rubber
bulb is not available, use a burette or a graduated
cylinder.
8. Keep volatile liquids (alcohol, ether, petroleum derivatives,
etc.) away from open flames.
9. The laboratory is a place of work. Playing around is wasteful
of time, equipment, and supplies.
10. For the safety of yourself and others, label all containers
which contain solid chemicals, liquids, etc.
"My uncle was a chemist
He isn't any more,
'Cause what he thought was H?0,
Was H2S04!" *•
11. Before leaving the laboratory, check your clothing for
spilled substances and thoroughly wash your hands.
B. Bacterial Studies
1. Treat all cultures as if they were pathogenic (disease
causing).
2. Plastic and rubber should not be autoclaved.
3. Stay with the autoclave while it is in operation. Turn it
off before you leave.
4. Keep work area and equipment sterile. This is necessary
from a health standpoint and to prevent contamination of
your cultures.
5. Disposal of cultures and resterilization of plastic petri
dishes can be accomplished as follows:
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Appendix 7
a. Using forceps remove the petri dish covers, and then
place covers, dishes, and cultures into a large beaker
or pan containing undiluted, liquid household bleach.
b. After 10 minutes remove the petri dishes, using tongs or
a rubber glove, and rinse them well under running water.
The wet pads and filter should be put into a plastic bag
and discarded.
c. Immerse the petri dishes and covers in a solution of
70% isopropyl (rubbing) alcohol for 10 minutes.
d. Remove the petri dishes and covers and stack them on a
clean surface. Assemble the dishes and covers. (They
are now ready for reuse.)
Chemistry
1. Tasting chemicals is dangerous and rarely leads to conclusive
results.
2. Should the occasion arise for smelling gases given off by
chemical reactions, waft the rising gases to your nose with
gentle sweeps of the hand.
3. To avoid sudden dangerous situations from occurring, plan
your activity in advance.
4. Always read and reread labels. Particularly note cautions
on labels.
5. When connecting rubber and glass apparatus, lubricate the
glass with water and assemble with a twisting motion.
6. Be careful when heating or cooling chemicals; be sure the
container is designed to be heated. Graduated cylinders
and bottles usually are not designed to withstand rapid
temperature changes.
Field Trips
1. Be sure to take a first aid kit on all field trips.
2. Wear appropriate clothing. Use old sneakers when wading
in unknown waters and swamps.
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Appendix 7
3. If you are going to be using boats in deep water, be sure
that everyone has a life jacket or other floatation device.
4. Pair off when working in situations where drowning could
occur.
5. Plan for emergency services by obtaining phone numbers of
appropriate services in the field area.
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US GOVERNMENT PRINTING OFFICE 197Z 514-146,'il 1-3
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