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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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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

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Appendix I
                  Nonmotile Green Algae: Coccoid
                          (Chlorophyceae)
                           Pediastrum
                Species of the Genus Scenedesimis
   S.  caudatus
S.  abundans
S. dimorphus
                          Plate III a.
                             A-77

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Appendix I
                  Nonmotile Green Algae: Coccoid
                          (Chlorophyceae)
                           Desmids
                                               go
   Closterium
Cosmarium
                         Staurastrum
                        Plate III b.
                           A-78

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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

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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

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Appendix I
            General Morphology of Algal Flagellates
Yellows
   Chromulina
Synura         Dinobryon          Lorica
Browns
           Peridinium                        Gyrnnodinuim



                          Plate VI b.
                            A-83

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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

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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

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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

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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

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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

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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

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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

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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

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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

-------
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

-------
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

-------
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

-------
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

-------
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

-------
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

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                                  -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

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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

-------
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

-------
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

-------
                                  -1-
STR-CLAS
100
110
120
130
140
150
160
170
180
190
200
210
220
230
240
250
260
270
280
290
300
310
320
330
340
350
360
370
380
390
400
410
420
430
440
450
460
470
480
490
500
510
520
530
540
550
560
570
580
590
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

-------
                                  -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

-------
                                  -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

-------
                                   -4-




STR-CLAS  (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
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

-------
                                   -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

-------
                                  -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

-------
                                   -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

-------
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.
                                 A-190

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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.
                                 A-191

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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.
                                 A-192

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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.
                                  A-193

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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).
                                  A-194

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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.
                                  A-195

-------
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.
                                  A-196

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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.
                                  A-197

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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:
                                  A-198

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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.
                                  A-199

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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
                                  A-200

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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.
                                  A-201

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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!!
                                  A-202

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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.
                                  A-203

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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
                                  A-204

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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
                                 A-205

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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.
                                  A-206

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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)
                                  A-208

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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.)
                                  A-209

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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.
                                  A-210

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Appendix 6
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.
                                   A-211

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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.)
                                   A-212

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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.
                                  A-213

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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.
                                   A-214

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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.
                                  A-215

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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.
                                   A-216

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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.)
                                  A-217

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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.
                                  A-218

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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.
                                  A-219

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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.)
                                  A-220

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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.
                                  A-221

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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.)
                                   A-222

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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)
                                  A-223

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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.
                                   A-224

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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.)
                                  A-225

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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.)
                                  A-226

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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.
                                  A-227

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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)
                                   A-228

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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.
                                  A-229

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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.)
                                  A-230

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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.
                                  A-231

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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.)
                                  A-232

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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.
                                  A-233

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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.)
                                   A-234

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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.)
                                  A-235

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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.)
                                  A-236

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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.
                                  A-237

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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.
                                  A-238

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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.)
                                  A-239

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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.
                                  A-240

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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)
                                  A-241

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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.
                                  A-242

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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.)
                                  A-243

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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)
                                  A-244

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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.
                                  A-245

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Appendix 6
VERTEBRATES  Animals that have an internal skeletal system.  (See
     Invertebrate.)
                                  A-246

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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.)
                                  A-247

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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.
                                  A-248

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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.
                                  A-249

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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:
                                  A-250

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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.
                                  A-251

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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.
                                        A-252
US GOVERNMENT PRINTING OFFICE 197Z  514-146,'il 1-3

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