EPA/600/3-85/044
                                                          June 1985
                          VALIDITY  OF EFFLUENT
                       AND AMBIENT  TOXICITY TESTS
                    FOR PREDICTING  BIOLOGICAL IMPACT,
                     SCIPPO CREEK,  CIRCLEVULE,  OHIO
                                 Edited by

                         Donald I.  Mount,  Ph.D.(a)
                                    and
                         Teresa J.  Norberg-King(a)
(a)  U.S. Environmental  Protection Agency.  Environmental  Research
     Laboratory — Duluth,  6201 Congdon Blvd. , Duluth,  Minnesota 55804.
                            U.S. Environmental Protection Apency
                            Region 5, Library  (PL-12J)
                            77 Wes! Jackson Doulevar.d,  12th Floor
                            Phiraon

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                      NOTICE

This document has been reviewed in accordance with
U.S. Environmental Protection Agency policy and
approved for publication.  Mention of trade names
or commercial products does not constitute endorse-
ment or recommendation for use.
                       11

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                          LIST OF INVESTIGATORS
                        LABORATORY TOXICITY TESTS
            Donald I. Mount(a) and Teresa J. Norberg-King(a)
               TIME-OF-TRAVEL STUDY-AND FLOW MEASUREMENTS
                           Jonathan C. Yost(b)
                          PERIPHYTIC COMMUNITY
                         Ronald J. Bockelman(b)
                   BENTHIC MACROINVERTEBRATE COMMUNITY
                          Michael T. Barbour(b)
                             FISH COMMUNITY
               David A. Mayhew(b) and David P.  Lemarie(b)
               COMPARISON OF LABORATORY TOXICITY DATA AND
                    RECEIVING WATER BIOLOGICAL IMPACT
            Teresa J. Norberg-King(a) and Nelson A.  Thomas(a)
PRINCIPAL INVESTIGATOR:  Donald I.  Mount,  Ph.D. (a)
(a) U.S.  Environmental  Protection Agency.   Environmental  Research
    Laboratory—Duluth,  6201  Congdon Blvd. ,  Duluth,  Minnesota 55804.
(b) EA Engineering,  Science,  and Technology,  Inc.  (formerly called
    Ecological Analysts,  Inc.),  Hunt Valley/Loveton Center,  15 Loveton
    Circle,  Sparks,  Maryland  21152.
                                  n

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                                CONTENTS


                                                                    Page

LIST OF INVESTIGATORS                                               in

LIST OF TABLES                                                      vii

LIST OF FIGURES                                                      ix

ACKNOWLEDGEMENTS                                                      X

FOREWORD                                                             xi

EXECUTIVE SUMMARY                                                   XIV

QUALITY ASSURANCE                                                   XVI

1.  INTRODUCTION                                                    1-1

2.  STUDY DESIGN AND SITE DESCRIPTION                               2-1

3.  LABORATORY TOXICITY TESTS                                       3-1

    3.1  Chemical and Physical Test Conditions                      3-1
    3.2  Results of Onsite Toxicity Testing                         3-2
    3.3  Results of Laboratory Testing—Duluth                      3-3
    3.4  Discussion and Conclusion                                  3-4

4.  TIME-OF-TRAVEL STUDY AND FLOW MEASUREMENTS                      4-1

5.  PERIPHYTIC COMMUNITY                                            5-1

    5.1  Chlorophyll a. and Biomass Measurements                     5-1
    5.2  Evaluation of the Periphytic Community                     5-2

6.  BENTHIC MACROINVERTEBRATE COMMUNITY                             6-1

    6.1  Community Composition                                      6-1
    6.2  Spatial Comparison of Key Taxa                             6-1
    6.3  Evaluation of the Benthic Community                        6-3

7.  FISH COMMUNITY                                                  7-1

    7.1  Community Structure                                        7-1
    7.2  Evaluation of the Fish Community                           7-2

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                            CONTENTS (CONT.)


                                                                    Page

8.  COMPARISON OF LABORATORY TOXICITY TEST DATA AND
    RECEIVING WATER BIOLOGICAL IMPACT                               8-1

    8.1  Predictions of Instream Community Impacts Based on
         Effluent Dilution Test and Ambient Toxicity Test Results   8-3
    8.2  Summary                                                    8-4

REFERENCES

APPENDIX A:  TOXICITY TEST AND ANALYTICAL METHODS

APPENDIX B:  HYDRCLOGICAL SAMPLING  AND ANALYTICAL METHODS

APPENDIX C:  BIOLOGICAL SAMPLING AND ANALYTICAL METHODS

APPENDIX D:  BIOLOGICAL DATA

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                             LIST OF TABLES
Number

 3-1       Mean dry weights and survival  for  fathead  minnow  larvae onsite
           effluent dilution test in receiving water.

 3-2       Survival and young production  for  Ceriodaphnia reticulata  in
           onsite effluent dilution test  in receiving water  and  ambient
           toxicity tests.

 3-3       96-hour percent survival  of  resident species exposed  to
           effluent concentrations,

 3-4       Mean dry weights and survival  for  fathead  minnow  larvae
           effluent dilution tests in two dilution water  types
           and shipped effluents.

 3-5       Survival and young production  for  Ceriodaphnia reticulata
           effluent dilution tests in two dilution water  types and
           shipped effluents.

 4-1       Measured flows on Scippo  Creek.

 5-1       Chlorophyll a, and biomass measurements  of  the  periphytic
           community,  Scippo Creek,  August  1982.

 6-1       Density and percent composition of the  most abundant  benthic
           macroinvertebrate species at each  sampling station, Scippo
           Creek,  August 1982.

 6-2       Shannon-Wiener diversity  indices and associated evenness
           and redundancy values calculated on benthic macroinvertebrate
           data,  Scippo Creek.

 7-1       Abundance of fish species by station, Scippo Creek,
           August  1982.

 C-1       Habitat characterization  of  the  sampling stations.

 D-1       Ranked  abundance listing  of  all  macroinvertebrates collected,
           Scippo  Creek,  August 1982.

 D-2       Number  of individuals and percent  composition  for benthic
           macroinvertebrates  collected,  Scippo Creek, August 1982.

 D-3       Analysis of variance and  Tukey's Studentized Range Test
           results for species of Chironcmidae,  Scippo Creek, August
           1982.

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                         LIST OF TABLES (Cont.)
Number                          Title

 D-4       Analysis of variance and Tukey's Studentized Range Test
           results for species of Hydropsychidae,  Scippo Creek,
           August 1982.

 D-5       Analysis of variance and Tukey's studentized range test
           results performed on species of Baetidae,  Scippo  Creek,
           August 1982.

 D-6       Results of a x2 test performed on the number of benthic
           macroinvertebrate taxa collected at each station.

 D-7       List of fish species and families collected,  Scippo Creek,
           August 1982.
                                  VT 1

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                             LIST OF FIGURES


Number                            Title

 2-1       Map of study site on Scippo Creek,  Circleville,  Ohio.

 4-1       Time-of-travel study on Scippo  Creek from Station 2  to
           Station 3.

 6-1       Diversity index and components  of the index in Scippo  Creek.

 6-2       Mean density of Chironomidae and Oligochaeta in Scippo Creek.

 6-3       Mean density of Trichoptera and Ephemeroptera in Scippo Creek.

 6-4       Mean density of Chironcmids in  Scippo Creek.

 6-5       Mean density of Trichopterans in Scippo  Creek.

 6-6       Mean density of Ephemeropterans in Scippo Creek.

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                            ACKNOWLEDGEMENTS

The assistance of the State of Ohio Environmental  Protection Agency staff
in locating the test site,  obtaining permission for location of the
mobile testing laboratory,  and for some onsite testing is acknowledged.
The plant maintenance staff was especially helpful with resolving elec-
trical problems.   The assistance of Thomas H.  Roush at the test site is
also gratefully acknowledged.   In addition,  the detailed technical review
provided by Charles Webster of the State of Ohio Environmental  Protection
Agency was greatly appreciated.

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                                FOREWORD


The Complex Effluent Toxicity Testing Program was initiated to support

the developing trend toward water quality-based toxicity control in the

National Pollutant Discharge Elimination System (NPDES) permit program.

It is designed to investigate, under actual discharge situations,  the

appropriateness and utility of "whole effluent toxicity" testing in the

identification, analysis, and control of adverse water quality impact

caused by the discharge of toxic effluents.


The four objectives of the Complex Effluent Testing Program are:
    1.  To investigate the validity of effluent toxicity tests in
        predicting adverse impact on receiving waters caused by
        the discharge of toxic effluents.

    2.  To determine appropriate testing procedures which will
        support regulatory agencies as they begin to establish
        water quality-based toxicity control programs.

    3.  To provide practical case examples of how such testing
        procedures can be applied to a toxic effluent discharge
        situation involving a single discharge to a receiving
        water.

    4.  To field test short-term chronic toxicity tests including
        the test organisms,  Ceriodaphnia reticulata and
        Pimephales promelas.
Until recently, NPDES permitting has focused on achieving technology-

based control levels for toxic and conventional pollutants in which

regulatory authorities set permit limits on the basis of national  guide-
lines.  Control levels reflected the best treatment technology available,
considering technical and economic achievability.   Such limits did not,
nor were they designed to,  protect water quality on a site-specific

basis.


The NPDES permits program,  in existence for over 10 years,  has achieved
the goal of implementing technology-based controls.   With these controls

largely in place,  future controls for toxic pollutants will,  of neces-

sity, be based on site-specific water quality considerations.

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Setting water quality-based controls for toxicity can be accomplished
in two ways.   The first is the pollutant-specific approach which
involves setting limits for single chemicals,  based on laboratory-
derived no-effect levels.   The second is the "whole effluent" approach
which involves setting limits using effluent toxicity as a control
parameter.  There are advantages and disadvantages to both approaches.

The "whole effluent" approach eliminates the need to specify  a limit  for
each of thousands of substances that may be  found in an effluent.   It
also includes all interactions between constituents as well as biological
availability.  Such limits determined on fresh effluent may not reflect
toxicity after aging in the stream and fate  processes change  effluent
composition.   This problem is less important since permit limits are
normally applied at the edge of the mixing zone where aging has not yet
occurred.

To date, eight sites involving municipal and industrial dischargers have
been investigated.  They are, in order of investigation:

    1.  Scippo Creek,  Circleville,  Ohio
    2.  Ottawa River,  Lima, Ohio
    3.  Five Mile Creek, Birmingham,  Alabama
    4.  Skeleton Creek, Enid, Oklahoma
    5.  Naugatuck River, Waterbury, Connecticut
    6.  Back River, Baltimore Harbor, Maryland
    7.  Ohio River, Wheeling, West Virginia
    8.  Kanawha River, Charleston,  West Virginia

This report presents the site study on Scippo Creek, Circleville,  Ohio,
which was conducted in August 1982.  The stream is small and receives
discharge from one industry.
                                   XII

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This project is a research effort only and has not involved  either
NPDES permit issuance or enforcement activities.
                                         Rick Brandes
                                         Permits  Division

                                         Nelson Thomas
                                         ERL/Duluth

                                         PROJECT  OFFICERS
                                         Complex  Effluent Toxicity
                                         Testing  Program
                                  xm

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                            EXECUTIVE SUMMARY

EPA recently issued a water quality-based policy which provides for
control of the discharge of toxic substances through the use of numerical
criteria and effluent toxicity limits in NPDES permits.   This policy is
the first broad scale effort to use effluent toxicity limits in the NPDES
permit program and a scientific basis for this approach is needed.

This report describes the first site study on Scippo Creek at Circle-
ville, Ohio,  which receives only one discharge from a chemical resins
plant using batch operations.   Scippo Creek is a small sunfish/bass
stream flowing through an agricultural area in central Ohio.   Previous
biological studies by the State of Ohio had shown measurable adverse
impact below the outfall and a grab sample of effluent tested before the
study indicated high toxicity.   Effluent dilution toxicity tests were run
with two test species both onsite and at a remote laboratory.   In addi-
tion,  toxicity tests were conducted onsite on ambient samples from four
river stations.  Biological studies were conducted at those stations and
included benthic macroinvertebrates,  fish, and periphyton.

The results of this study revealed no biological impact  in the stream
except for a small area of changed species composition at the outfall
which is presumed to be caused by a physical change in the substrate
from settled precipitate which clogged the sediment interstitial spaces.
No toxicity to C_. reticulata.  fathead minnows,  or resident species was
measured in the 100 percent effluent.

The processed waste is held in a detention tank after treatment.  Several
times each week the tank is pumped and treated waste is  discharged.   The
initial grab sample of effluent was apparently taken when process waste
was being discharged, but the composite sampling process used in this
study reduced peak concentrations.  Importantly, the composite sample
toxicity results best predicted the lack of community impact.   New treat-
ment equipment had been installed after a previous biological  survey
which was conducted by Battelle Laboratories (1971).   Operation of this
                                   xiv

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new equipment may have improved waste treatment and presumably that is
why little or no effect was found in the stream.   Correctly predicting
no impact to a receiving stream is a requirement of tests used for regu-
latory purposes.
                                   xv

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                            QUALITY ASSURANCE

Coordination of the various studies was completed  by  the  principal
investigator preceding and during the onsite work.  A reconnaissance
trip was made to the site before the study and  necessary  details  regard-
ing transfer of samples,  specific sampling sites,  dates of  collections,
and measurements to be made on each sample were delineated.   The  eve-
ning before the study began,  a meeting was held onsite to clarify again
specific responsibilities and make last minute  adjustments  in schedules
and measurements.   The mobile laboratory was established  as the center
for resolving problems and adjusting of work schedules as delays  or
weather affected the completion of the study plans.   The  principal
investigator was responsible for all Quality Assurance-related deci-
sions onsite.

All instruments were calibrated by the methods  specified  by the manu-
facturers.  For sampling and toxicity testing,  the protocols described
in the referenced published reports were followed.  Where identical mea-
surements were made in the field and laboratory, both instruments were
cross-calibrated for consistency.
                                    xvi

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                            1.  INTRODUCTION

This study was the first site investigated in the Complex Effluent
Testing Program.  The site was chosen because the stream was  small
and effluent-dominated by one discharge.   Equipment and  methods  had
been untried for onsite testing and the mobile laboratory had just been
assembled.  Many logistical and procedural details had to be  developed
before more complex sites could be attempted.   Special emphasis  was
placed on improving test procedures and simplifying equipment needs,
as well as meeting the major objective which was  to use  toxicity tests
to predict expected biological impact in  the stream.

This report is organized into sections corresponding to  the project
tasks.   Following an overview of the study design and a  summary  of the
description of the site,  the chapters are arranged into  toxicity testing,
hydrology, and ecological surveys.   An integration of the laboratory  and
field studies is presented in Chapter 8.   All methods and support data
are included in the appendixes for reference.
                                   1-1

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                  2.  STUDY DESIGN AND SITE DESCRIPTION

 The effluent evaluated was from a plastics resin plant in central Ohio
 that discharged to a small stream in a flat, rich agricultural area.
 There were no other known discharges to the stream.   The influent was
 taken from a well and most of the discharge was cooling water.  The
 process water was treated in rotating biological contactors and held in
 tanks capable of holding the waste volume generated in 30 days.  Several
 times each week, the treated waste was pumped into the cooling water
 discharge and then discharged into Scippo Creek.  The temperature of
 the discharge was considerably cooler than the stream at the time of  the
 study in July-1982.  There was a substantial amount of precipitate from
 the well water observed below the outfall.

 Study components included 7-day Ceriodaphnia reticulata toxicity tests
 on samples from each of four river stations and various concentrations
 of the effluent; 7-day larval growth tests on fathead minnows in vari-
 ous concentrations of the effluent;  tests of indigenous species; ambient
 toxicity caging studies; time-of-travel analysis for the effluent; and
quantitative assessment of the benthic macroinvertebrate,  periphytic,
 and fish communities.   The study was conducted 9-16  August 1982.

 The study area on Scippo Creek was located above the confluence with
 the Scioto River.   Scippo Creek (Figure 2-1) is shallow (less than 0.6 m
 in depth) and 10-20 m in width at the study area.   Pool areas predominate
with periodic riffle sections along  its length.   The study area incor-
 porated 6.7 river kilometers (RK)  of stream and five sampling locations.
 Habitats sampled were riffles and  pools for benthic  macroinvertebrtes  and
a combination of both for fish.   Periphyton samples  were taken from run
areas  or pools where available.   The station locations as  depicted in
Figure 2-1  are:
       Station 1—0.28 km upstream of  the  effluent  outfall.
       The sampling station was  located  in a  straight  stretch,
       approximately 20 m in length, downstream  of  a bend in  the
       creek.   The station was shaded  approximately 80  percent
       by deciduous canopy.   Stream width  was approximately 15 m.
       The riffle substrate consisted  of pebble-cobble, with
                                   2-1

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       varying amounts of sand deposited among the rocks.  The
       substrate of the pools was primarily sand, with small
       amounts of mud.  The pools were relatively free of debris
       but did contain some leaf packs.

       Station 2—0.1 km downstream of the outfall.  The station
       was located approximately 10m downstream of a slight bend
       in the creek.  Shading was provided by a deciduous canopy
       which covers about 40 percent of the station.   Stream
       width was about 10m.  The substrate of the riffle con-
       sisted of pebble and gravel, with seme cobble overlying
       bedrock.   Pools were deepest at this station and contained
       some debris (e.g., logs,  branches) and leaf packs.

       Station 3—1.3 km downstream of the outfall.  The sampling
       station had little canopy cover (less than 25 percent) and
       was approximately 15 m in width.  Substrate of the riffle
       area was primarily pebble and gravel, with pockets of sand.
       The pools had sand-mud bottoms and contained a fallen tree.

       Station 4—3.7 km downstream of the outfall and immedi-
       ately downstream of the U.S. Rte.  23 bridge.  Canopy cover
       at Station 4 was near 100 percent.   The creek width was
       approximately 20 m.  The riffle consisted of cobble and
       pebble substrate with some sand.  The substrate of the
       pools was sand with little debris.

       Station 5—5.3 km downstream of the outfall and immediately
       downstream of the confluence with Congo Creek.   The canopy
       cover at Station 5 was about 90 percent.   The riffle sub-
       strate was cobble and pebble overlying bedrock.   Some sand
       pockets were also present.   The width of the creek at this
       station was approximately 20 m.  The pools were sand sub-
       strate and free of debris.
See Table C-1 for a pool vs.  riffle habitat description at the sampling

locations.


Temperature, dissolved oxygen,  specific conductance,  and pH were moni-

tored during biological collections and the first half of the fish caging

study.  The instruments used for water quality measurements were a Hydro-

lab Model 1041, a YSI Model 57  Dissolved Oxygen Meter,  and a YSI Model 33

Salinity-Conductivity-Temperature Meter.  Dissolved oxygen ranged from

7.9 to 13.6 mg/liter, with many readings above 100 percent saturation.

The pH ranged from 7.5 to 8.5.
                                   2-2

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The temperature effect from the discharge was variable,  sometimes causing
virtually no change in receiving water temperature,  and  at other times
decreasing the temperature at Station 2 by 8 C.   Although diel  tempera-
ture patterns were not studied,  the water had returned to normal temper-
ature range at Station 4.

At Station 1, conductivities from 550 to 590 /umhos were  recorded over
10-13 August.  However, the discharge caused rapid,  large variations in
conductivity downstream.   One such event occurred on 12  August  at Station
2, when conductivity increased from 806 to 1,229  ^mhoa in 10 minutes, and
to 1,535 Mmhos 30 minutes later.   Approximately 8 hours  later,  a reading
of 630 /jjnhos was recorded, and 2,390 jumbos was measured  the following
day.  At Station 3, the conductivity ranged from  720 to  1,170 Mmhos  and
was fairly stable at Stations 4  and 5 with ranges of 640-700 and 640-680
Mmhos,  respectively.
                                   2-3

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Figure 2-1. Map of study site on Scippo Creek, Circleville, Ohio.



                          2-4

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                      3.  LABORATORY TOXICITY TESTS

Laboratory toxicity tests using Ceriodaphnia reticulata, fathead minnows,
and resident species were conducted to determine the maximum effluent
concentrations that would not have chronic toxicity, and to measure the
ambient toxicity before and after the effluent is discharged in order
to estimate the persistence of the toxicity (Stations 1-4).   Several  sub-
sidiary objectives were also pursued.   Samples of effluent were shipped
to Duluth to determine if shipping and delayed testing would produce
different results from those of onsite testing.   Additional  tests at
Duluth were done in Lake Superior water to see what effect a different
dilution water might have on the results.   Descriptions of the toxicity
test methods are presented in Appendix A.

Animals from eight different families found in the stream were tested
onsite to see if the resident organisms were more or less sensitive
than the laboratory animals.  If there were differences,  the acceptable
effluent concentration (AEC) for the resident species could be estimated
by dividing the acute/chronic ratio into the LC50 values of  the resident
species.

Another toxicity test procedure was used with bluntnose  minnows
(Pimephales notatus).  The minnows were caged and set at Stations 1-4.
Due to infection,  difficulties in capture and handling,  and  effects of
lower water temperatures in the effluent,  the test  results are regarded
as invalid and have not been presented.

3.1  CHEMICAL AND PHYSICAL TEST CONDITIONS

In the onsite tests,  the dissolved oxygen (DO)  concentration in fathead
minnow and resident species tests ranged from 4.6 to 7-7  mg/liter,  as
measured at the end of each 24-hour period.   Initially,  DO was very near
saturation.   In the Ceriodaphnia reticulata tests,  DO ranged from 6.6 to
8.0 mg/liter.   The pH in all tests was  from 7.5  to  8.2.   Temperature for
the fathead minnows and resident species was from 18.5  to 25 C,  and was
25±1  C C)  for £.  reticulata.
                                   3-1

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In the shipped-effluent tests conducted in Duluth,  Minnesota,  DO was
4.8-7-9 and 4.8-7.2 nig/liter for the lake and receiving water tests,
respectively.  The pH was 7.7 and 8.4 in the lake and receiving water,
respectively.  Water temperature was maintained at between 24 and 26  C.

Receiving water ranged from 300 to 310 mg/liter hardness (as  CaCO_)
before effluent was added,  and up to 400 mg/liter in high effluent con-
centrations.  Corresponding values for alkalinity were 250-260 and 324
mg/liter,  respectively.

3.2  RESULTS OF ONSITE TOXICITY TESTING

Table 3-1  contains the data from the larval  growth  test with  fathead
minnows (Pimephales promelas) exposed to various effluent concentrations
diluted with receiving water and tested onsite.   The weights  are actual
values for each replicate and the treatment  mean is a weighted average
of the replicate means. There was no significant difference in survival
or weights at any effluent concentration.  Fathead  minnow weights were
slightly higher at the 25 and 100 percent effluent  exposure,  perhaps
attributable to the additional food in the effluent.   The statistical
analyses for the weight and survival data are described in Appendix  A.

Data from  the onsite tests with jC.  reticulata.  using the effluent dilu-
tion test  and the ambient toxicity test,  are shown  in Table 3-2.   The
results in both tests,  and especially in those  test solutions with no or
low effluent concentrations,  are heavily influenced by a fungal  growth in
the test containers that entrapped the animals  and  prohibited  swimming.
Although the entrapped animals lived for several  days and produced some
young,  their development was impaired and the test  results are not useful
in evaluating direct toxicity.  When the animals were transferred each
day,  they  were dislodged from the growth by  directing a jet of water
from the eye dropper and considerable force  was needed to free them.
They soon became entrapped again because the fungal  growth would develop
in a few hours although the beakers were thoroughly brushed during wash-
ing,  and rinsed before reuse.   In the ambient test,  the fungus problem
was worse  at Station 1  above the outfall and diminished downstream which
                                   3-2

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suggests that the fungal growth was not caused by the effluent.   Young
production from surviving females was not significantly different among
stations (Table 3-2).  As a result of the fungus, survival was not
concentration-dependent and, therefore,  any effluent-caused mortality
cannot be ascertained.

Table 3-3 contains the resident species data.  One of the fish species
tested, Pimephales notatus. died of a fungal infection within the first
24 hours.  Survival between exposure concentrations was similar for the
remaining seven species, and generally varied from 40 to 100 percent for
all seven genera.  Lowest survival was observed in the middle concentra-
tions.  Mortalities could not be attributed to effluent toxicity, only
to handling.

3.3  RESULTS OF LABORATORY TESTING—DULUTH

The survival and growth data for larval  fathead minnow growth tests,
conducted at the Environmental  Research Laboratory in Duluth,  Minnesota,
with receiving water and Lake Superior water, are given in Table 3-4.
There were no significant differences observed for either the growth
or survival data for the receiving water dilution test.   Survival was
generally lower in the Lake Superior dilution water than in the  receiving
water test.   However,  there were no significant differences for  survival
or growth in any effluent concentrations with Lake Superior water as the
diluent.

The data for .C.  reticulata reproduction and survival  in various  concen-
trations of effluent and two diluent waters are presented in Table 3-5.
In the receiving water test,  none of the exposure groups were signifi-
cantly lower than the control for either reproduction or survival.   In
the Lake  Superior water test,  survival was also not significantly lower
between concentrations.   All exposure groups at concentrations of 5
percent effluent,  and above,  had significantly higher (P 1 0.05)  young
production which may be a result of additional food.
                                   3-3

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The fungal growth that entrapped the test animals in onsite  tests did
not occur in the tests done at Duluth,  Minnesota,  in either  dilution
water.  The reason for this is unknown.   The results of  these  shipped-
effluent tests are considered valid for evaluating toxicity  of the
effluent because control survival was acceptable.

3.4  DISCUSSION AND CONCLUSION

The results of the tests using fathead minnows and £.  reticulata indicate
no adverse chronic effect even at 100 percent effluent.   The resident
species tests gave no evidence of acute toxicity  nor did the shipped-
sample tests with the standard species.   Based on these  data,  no effect
of the discharge on Scippo Creek would be expected,  even close to the
point of discharge.   Visual inspection of the discharge  area revealed
yellow-orange deposits of precipitate which might cause  a physical
effect, especially on the benthic organisms.   For those  species able
to utilize the increased microorganism population associated with the
effluent,  a beneficial effect might be expected.

The resident species tests were unsatisfactory because of handling mor-
tality.  If such species are to be tested,  a suitable  acclimation period
must be provided.   In addition,  for those species that live  in flowing
water, a water current should be provided in the  test  chamber.   However,
despite these considerations,  it can still be concluded  that the 100
percent effluent was not toxic to resident species.
                                   3-4

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   TABLE 3-1  MEAN DRY WEIGHTS AND SURVIVAL FOR FATHEAD MINNCW LARVAE
              ON SITE EFFLUENT DILUTION TEST IN RECEIVING WATER
Larval Weight (ing)
Percent Effluent (
Replicate
A
B
C
D
Weighted mean(a)
SE(b)


Replicate
A
B
C
D
100
0.17
0.22
0.37
0.20
0.238
0.034


100
100
90
90
90
_J5_
0.16
0.22
0.37
0.20
0.240
0.034
Percent

_25_
70
100
90
100
10
0.18
0.16
0.22
0.19
0.187
0.033
Survival
Percent
10
90
100
90
100
• -5-
0.25
0.22
0.22
0.21
0.225
0.033

Effluent (

100
90
100
100
v/v)
-L~
0.29
0.20
0.18
0.18
0.213
0.033

v/v)
— I—
100
100
100
90

Control
0.30
0.21
0.12
0.11
0.192
0.035


Control
100
90
60
100
Mean
93
90
95
98
98
(a)  Mean for the group of four replicates,  calculated as a weighted
     mean.
(b)  Standard error of the weighted means.
                                  3-5

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  TABLE 3-2  SURVIVAL AND YOUNG  PRODUCTION  FOR Ceriodaphnia reticulata
             IN THE ON SITE EFFLUENT DILUTION  TEST  IN  RECEIVING  WATER
       	AND FOR AMBIENT TOXICITY TESTS(a)	___	
                          Receiving Water Test
   Percent
Effluent (v/v)

Control
  1
  5
 10
 25
100
Percent
Survival

  30
  60
  70
  70
  80(b)
  40
Mean Number
 of Broods

   3.0
   2.3
   3.0
   3.0
   3.0
   2.8
Mean Number
 of Young
Per Female
   14,
   10
   13
   15.
   14,
   13.3
95 Percent
Confidence
Intervals

 9.8-19.0
 6.5-14.9
 7.2-18.6
11.7-18.7
10.8-18.0
 5.2-21.2
                           Ambient Stream Test
Station

   1
   2
   3
   4
Percent      Mean Number
Survival      of Broods

   10           3.0
   60           3.0
   50           2.2
   60           3.2
                Mean Number
                 of Young
                Per Female

                   13.0(c)
                   14.8
                   12.8
                   17.5
                95 Percent
                Confidence
                Intervals

                   —(d)
                12.6-17.0
                 7.6-18.0
                14.3-20.6
(a)  The results were affected by fungal  growth  in the  test  containers
     which entrapped the Ceriodaphnia reticulata.   Organism  development
     was impaired and control  mortality was  high so these  results  are not
     useful in evaluating direct  toxicity.
(b)  Survival was significantly higher than  control (P  10.05).
(c)  Mean number of young per  single surviving adult..
(d)  Confidence intervals were not calculable due  to the small  sample
     size of surviving females.   See Appendix A  for description of
     statistical analysis.
                                   3-6

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   TABLE 3-3  96-HOUR PERCENT SURVIVAL OF RESIDENT SPECIES EXPOSED TO
              EFFLUENT CONCENTRATIONS
Percent Effluent (
Test Oreani
Etheostoma
Qrconectes
Hydroosvche
(a)
sins
sp.
sp.
sp.
Heptageniidae
Philopotamidae
Ancylidae
Psephenidae


100
100
100
60
40
40
100
100
100
100
60
80
60
100
80
'v/v)

50 25
100
100
80
60
40
80
100
100
100
80
80
20
100
60
100
100
80
40
20
100
80
100
100
80
60
40
100
100
1


Control
A^
00
100
1


1
1
00
40
80
00
00
100
100
80
80
40
100
80
(a)  One of the species tested,  Pimephales notatus.  died of a fungal
     infection within 24 hours of test initiation.

Note:  A and B represent replicate test results.
                                   3-7

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TABLE 3-4   MEAN  DRY WEIGHTS  AND SURVIVAL  FOR FATHEAD  MINNCH  LARVAE
             EFFLUENT DILUTION  TESTS  IN WO DILUTION WATER TYPES AND
             SHIPPED EFFLUENTS
Receiving .
Water ra;
A
B
C
D
Weighted mean(b)
SE(o)
Receiving .
Waterra;
A
B
C
0
Mean
Lake Superior
Water
A
B
C
D
Weighted mean(b)
SE(o)
Lake Superior
Water
A
3
C
D
Mean
Larval Weight fmg)
Percent Effluent
100
0.58
0.40
0.4?
0.52
0.495
0.026
25 10
0.42 0.47
0.50 0.48
0.52 0.49
0.48 0.35
0.482 0.458
0.029 0.030
0.48
0.29
0.38
0.39
0.393
0.031
Peroent Survival
Percent Effluent
100
90
80
90
90
88
..25_ 10
90 100
100 100
100 100
100 60
98 90
100
70
90
90
88
( v/v)
0.42
0.46
0.39
0.41
0.421
0.029
(v/v)
100
100
90
100
98

Control
0.42
0.50
0.54
0.51
0.466
0.029

Control
100
100
90
100
98
Larval Weight (ing)
Peroent Effluent (v/v)
100
(d)
(d)
(d)
(d)
0.495
0.026

25 . 1Q_
0.48 0.28
0.44 0.45
0.49 0.39
0.44 0.37
0.461 0.378
0.025 0.025
Percent Survival
0.40
0.44
0.43
0.46
0.434
0.027

Percent Effluent (
100
90
30
90
90
38
._ 2-; _ 10
90 70
100 90
90 90
100 90
95 85"
80
100
90
100
93
0.33
0.32
0.25
0.301
0.026
'v/v)
50
80
90
80
75
Control
0.30
0.45
0.42
0.36
0.381
0.031

Control
60
60
60
60
60
    (a)  Fron Scippo Creek.
    (b)  Mean for the group of four replicates, calculated as a weignted
         mean.
    (o)  Standard error of the weighted aeans.
    (d)  The 100 percent effluent test was conducted once.  The data are
         provided under the receiving water teat data.
                                     3-8

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  TABLE 3-5
EFFLUENT DILUTION TESTS IN TWO DILUTION WATER TYPES AND
SHIPPED EFFLUENTS

Percent
Effluent

Control
1
5
10
25

Control
1
5
10
25
100

Percent
Survival

90
100
100
80
60

90
100
100
100
90
100

Mean Number
of Broods
Lake Superior Water
2.8
2.9
2.9
3.0
3.0
Scippo Creek Water
3.0
3.0
2.9
3.1
3.0
3.0
Mean Number
of Young
Per Female

14.4
18.2
19.6(a)
22.3(a)
21.2(a)

20.6
20.0
20.5
21.0
21.8
21.2
95 Percent
Confidence
Intervals

10.9-17.8
15.8-20.6
17.9-21.3
19.4-25.3
21.4-24.5

19.5-21.6
18.0-22.0
18.1-22.9
18.2-23.8
19.9-23.6
20.2-22.2
(a)  Mean is significantly greater than the control mean (P 1 0.05).
                                  3-9

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             4.   TIm-OF-TRAVEL STUDY AND FLOW  MEASUREMENTS

The objective of the hydrology study in Scippo  Creek was  to  ascertain
time-of-travel for the effluent,  from the discharge  to the end  of  the
study area.   Two complementary tasks were performed:  flow measurements
at the biological stations (10 and 13 August  1983),  followed by the
release of dye and subsequent monitoring of its passage downstream
(10 August 1983).  The sampling and analytical  methods of the
hydrological data are presented in Appendix B.

The average cross-sectional velocity from a flow measurement is
physically different from a dye study velocity  measurement.   The flow
measurement represents the average velocity through  a specific  cross-
section and is dependent on the cross-sectional area.   In contrast,
the dye study velocity represents an actual time-of-travel between two
points and is more representative of average  conditions over a  reach.

The results of the dye monitoring at Stations 2 and  3 are shown in
Figure 4-1.   Following release of the Rhodamine WT dye (1330 hours)
in the effluent, the leading edge of the dye reached Station 2  at  1426
hours and the peak of the dye distribution (a concentration  of  207 PPb),
occurred at 1432 hours.  At Station 3, located  1.2 km farther downstream,
the leading edge was observed at 1645 hours.   The peak dye concentration
(37.5 Ppb) arrived at 1735 hours.  The dye samples collected at Station 4
(1845-2245 hours) showed no dye above background level.  The observed
time interval for the peak dye concentration to pass from Station  2  to
Station 3 yields an average velocity for this section of  Scippo Creek  of
11 cm/sec.

Table 4-1 presents the flows and average cross-sectional  velocity
measured at the biological sampling stations.   On 10 August,  a  flow  of
0.033 m3/sec was measured upstream of the discharge.  The average  of the
three downstream flows was 0.107 tn3/sec.   The flow difference measured
between Stations 1 and 2 of 0.100 m3/sec (2.3 mgd) is consistent with  the
nominal reported discharge flow of 2.5 mgd (0.109 m3/sec).  The average
velocity calculated from the dye study of 11  cm/sec  is more  similar  to
                                   4-1

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the measured velocities at Stations 4  and 5  than at Stations 2 and 3.
The higher velocities measured  at  Stations 2  and 3 (31.1 and 22.9 cm/sec,
respectively) appear to be associated  with narrower river widths.  They
are not representative of  that  portion of the river.  Using the velocity
of 11 cm/sec resulting from the time-of-travel  study,, the peak dye dis-
tribution would have been  expected at  Station 4 at 2345 hours.  Since
sampling stopped at 2245 hours,  the leading  edge of the dye at Station 4
was probably not sampled.

The time-of-travel study velocity  of 11 cm/sec  is equivalent to an
exposure time of 2.5 hours for  each kilometer of movement downstream of
the average water parcel from the  point of discharge.  Water parcels in
the leading edge of the distribution would have experienced an exposure
time of less than average, whereas parcels in the tail of the distribu-
tion would have had longer exposure times.   Between Stations 2 and 3> the
leading edge of the dye distribution traveled at 14.43 cm/sec, which is
equivalent to 1.9 hours of exposure time for  each kiloneter downstream.
Thus, it would be expected that at a 1-km station, the average exposure
time is 2.5 hours, with the majority of water parcels having an exposure
between 1.9 and 3 hours.  At a  2-km station,  the average exposure time is
5 hours, with the majority of water parcels  having an exposure between
3.8 and 6 hours.

The longitudinal dispersion coefficient for  a flow channel (units of area
divided by time) is a measure of the rate of  the spatial expansion of a
group of water parcels with respect to its center of mass.  The center of
mass moves downstream at the average stream  velocity, whereas individual
parcels disperse due to turbulence, velocity  gradients, and associated
phenomena in natural streams.  Using Equations  B-2 and B-3, the longi-
tudinal dispersion coefficient  for Scippo Creek is  17.7 m2/min.
                                   4-2

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                                                                 o
                                                                 o
                                                                 O5
                                                                 O
                                                                 o
                                                                 CO
                                                                        
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                TABLE 4-1   MEASURED FLOWS ON  SCIPPO  CREEK
Station Date
1 10 AUG 1982
2 10 AUG 1982
3 10 AUG 1982
M 10 AUG 1982
4 13 AUG 1982
5 13 AUG 1982
Flow(a)
(nr/sec)
0.033
0.133
0.102
0.086
0.080
0.120
Average
Velocity
(cm/ sec)
5.5
31.1
22.9
7.3
11.3
9.1
(a)  Obtained from measured  velocities and the  cross-sectional area
     of the creek at each  station.
                                  4-4

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                        5.   PERIPHYTIC COMMUNITY

The study investigated the periphytic community by measuring  chloro-
phyll a. and biomass.   The relatively short reproduction time  and rapid
growth of periphytic algae result in quick response  to  changes in water
quality.   A change in the periphytic community may be either  a reduc-
tion of an important habitat or food source for other organisms or  the
enhancement of nuisance species of algae (that neither  support lower
trophic levels nor are aesthetically pleasing).

5.1  CHLOROPHYLL a AND BIOMASS MEASUREMENTS

The samples for chlorophyll a and biomass analyses were collected on
12 August 1982 from Stations 1 through 4.  The samples  contained large
amounts of sediment and flocculant material,  except  at  Station 1.   Due to
excessive silt, replicates 2A and 4C had to be discarded.

Chlorophyll a values ranged from 16.4 to 330.0 mg/m2.   Both of these
extreme values were from Station 3 (Table 5-1).   This substantial range
in values may be caused by changes in natural stream conditions, habitat
availability, or sampling conditions.   Mean chlorophyll s, values ranged
from 44.7 to 131.7 mg/m2 at the four stations.  The  upstream  station
(Station 1) and the farthest downstream station sampled for periphyton
(Station 4) had similar mean values for chlorophyll  a:   38.1  and 39.2
mg/m2, respectively.   Mean chlorophyll a values at Station 2  averaged
129.7 mg/m2.  Station 3 averaged 131.7 mg/m2.  Results  of Analysis  of
Variance (ANOVA) demonstrated that there was no difference among stations
when all data were considered,  versus a significant  difference (P 1 0.05)
among stations when Station 3 chlorophyll a values were omitted.

Periphyton biomass was lowest at Station 1  and highest  at Station 2.
Station 1 had a mean biomass,  measured as ash-free dry  weight (AFDW),
of 19.9 g/m2. Station 2 had a mean of 70 g/m2 AFDW.  Mean periphyton
biomass at Station 3 decreased by a factor of 1.7 from  Station 2, and
averaged 40.9 g/m2 AFDW.  Periphyton biomass was lower  at Station 4,
where the average was 28.2  g/m2 AFDW.   Station 3  had the largest range
                                   5-1

-------
between replicates (4.6-107.0 g/m2), with the highest and lowest AFDW.
Chlorophyll a and AFDWs are measures of algal biomass.  Since analyses
for these parameters  were from  the  same samples, similar results between
replicates would be expected.   Results of ANOVA indicated that there was
no significant difference in AFDW between stations when all data were
considered.  However, when Station  3 data were emitted, very significant
differences (P  below  the discharge, suggests enrichment
although within-station (replicate) variation was high, especially at
Station 3.  A similar trend of  increasing biomass was noted but was
probably due to a combination of periphytic and non-algal constituents.
However, no identifications were made  to ascertain the composition of
the periphytic community.
                                   5-2

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        TABLE 5-1  CHLOROPHYLL a AND BIOMASS MEASUREMENTS OF THE
                   PERIPHYTIC COMMUNITY. SCIPPQ CREEK.  AUGUST 1Q82

Chlorophyll .a
2
Station/ Replicates (me/m )
1 A
B
C
D

2 A
B
C
D

3 A
B
C
D

4 A
B
C
D
40.6
77.0
26.7
34.6
Mean 44.7
-(c)
185.0
61.2
143.0
Mean 129.7
152.0
330.0
16.4
28.5
Mean 131.7
37.0
28.5
— (c)
52.1
Biomass(a) Autotrophic
2 (b)
(e/m ) Index
24.6
29.5
12.4
13.0
19.9 522
— (c)
97.1
42.7
70.6
70.1 540
41.2
107.0
4.6
10.6
40.9 311
26.4
19.1
— (c)
39.2
                     Mean
39.2
28.2
719
(a)  Ash-free dry weight.
(b)  Weber 1973.
(c)  Sample rejected because  of  excessive  sediment  load.
                                   5-3

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                 6.  BENTHIC MACROINVERTEBRATE COMMUNITY

This survey investigated the benthic community in Scippo Creek.   Samples
were collected at five stations.  Because of the relatively low degree
of mobility, the benthic community is considered to be a good indicator
of response to adverse conditions at specific locations.  The degree of
community stability within the study areas can be measured by comparing
composition and dominance.  An alteration in community structure,  stand-
ing crop, or species composition of the benthos,  beyond the limits of
normal fluctuation within the receiving waterbody, would be regarded
as an adverse effect.   Increased abundance of nuisance insect larvae
or other benthic species also would be regarded as adverse effects.

A description of the sampling and analytical methods is presented  in
Appendix C.   Supportive data are summarized in Appendix D.

6.1  COMMUNITY COMPOSITION

The benthic community of riffle habitats in Scippo Creek comprised 104
taxa of which only 20 contributed ^.1  percent to the community population
(Table 6-1).  Of the 104 taxa collected during August,  only two  macro-
invertebrates,  Chironomus spp.  and Cricotopus tremulus (both midges),
constituted greater than 10 percent of the benthic fauna.   Six insect
taxa composed greater than 50 percent of the fauna,  suggesting that,
although the benthic community is diverse in variety of taxa,  the  struc-
ture of the community is dominated by relatively  few insect species.   Of
the 20 taxa and life stages composing one percent or more of the benthos,
12 are in the Chironcmidae family.   This midge-dominated community is
present at all stations.

6.2  SPATIAL COMPARISON OF KEY TAXA

Community diversity data based on number of taxa  and abundance of  indi-
viduals within taxa show that diversity was lowest at Station 2  and
similar at the other stations (Table  6-2).   Conversely,  evenness,  which
compares relative distribution of individuals within taxa  among  stations,
                                   6-1

-------
was also lowest at Station 2.   Redundancy, which reflects relative
dominance of taxa, was highest  at Station 2.  These community differences
at Station 2 were the consequence of  the lowest number of taxa (43 taxa)
and the greatest abundance of specimens (17,761 organisms/m2).

Figure 6-1 illustrates this pattern of decreasing diversity at Station
2 and increase at Station 3 to  a  value similar to that noted at Station
1.  The number of taxa also decreases from Station 1 to its lowest point
at Station 2, increases at Station 3, decreases again at Station 4.  It
then increases to a maximum of  70 taxa at Station 5.  A \2 test was used
to test for differences in the  number of taxa encountered at each station
compared to the expected composition  of the reference station.  The
results of this test indicated  that the lower number of taxa encoun-
tered at Station 2 was significantly  different (P £ 0.05) from the number
of taxa at Station 1 (Table D-6). The number of taxa at Stations 3, 4,
and 5 was not significantly different from the control.  The total number
of organisms at each station follows  a pattern of low density at Station
1, an increase to peak abundance  at Station 2, followed by a steady
decrease at the downstream stations to a density at Station 5 similar to
that at Station 1.

The community at Station 2 was  dominated by two taxa, each of which
composed more than 20 percent of  the  benthos (Table 6-1), whereas no
taxon constituted more than 20  percent of the benthos at other stations.
The overwhelming dominance of Chironomus spp. and Cricotopus tremulus
at Station 2 was not found at any other station.  The dominance of these
taxa at Station 2 was responsible for the lower diversity index at that
station.

Chirononidae and Oligochaeta were present in peak densities at Station 2.
They composed 93 percent of the benthos at that station (Figure 6-2).
Both groups steadily decreased  in abundance downstream.  In contrast,
Trichoptera and Ephemeroptera decreased from Station 1 to their lowest
densities at Station 2 and then increased at downstream stations (Figure
6-3).  The chironcmid abundance trend was primarily due to three taxa—
Chironomus spp., Cricotopus tremulus. and Polypedilum convictum—all
                                   6-2

-------
similarly distributed among stations, although at different abundance
levels (Figure 6-4).  Only at Station 4 were two of these species—
£. tremulus and £. convictum—not found.  Results of a one-way Analysis
of Variance (ANOVA) and Tukey's Studentized Range Test performed on these
three chironomid taxa indicated that the greater densities at Station 2
were highly significantly different (P = 0.0001) from densities at other
stations (Table D-3) .  For .P. convictum. the densities at Stations 2 and
3 were not significantly different.  No significant differences in abun-
dance were found among Stations 1,4, and 5 for all three species.   The
high abundance of midges at Station 3 was caused primarily by genera not
present in abundance at other stations.  Two of these midges—Cricotopus
trifascia and Rheotanytarsus spp.—were not found at any other station.
Par at any tarsus spp. was uncommon,  except at Station 3 (Table 6-1).

Cheumatopsvche spp. and Hydropsvche spp. are the dominant trichopterans
in the study area, reflecting the abundance trend of the group among
stations (Figure 6-5).  Results of the ANOVA and Tukey's test performed
on Hvdropsyche spp., Cheumatopsvche spp., and early instar Hydropsychidae
indicated that lower densities at Station 2 were very significantly dif-
ferent (P = 0.0001, 0.0001, and 0.009, respectively) from those at other
stations (Table D-4).  However,  overlap in the station means (natural
log-transformed) indicates that distinct station differences in the
early life stage of Hydropsyche larvae were not apparent in August  1982.
Baetis spp. is the numerically dominant mayfly in the study area and,
with the early instars,  accounts for the abundance of the mayfly group
(Figure 6-6).   Although densities of Baetis spp.  were very significantly
different (P = 0.0031) among stations (Table D-5), the Tukey's range test
results exhibited overlap of station means.   No significant differences
in the distribution among stations were found with early instar Baetidae
(Table D-5).   Differences in abundance between the caddisflies and  may-
flies are the  reversed abundance peaks at Stations 3 and 4.   Both groups
decreased in numbers at Station 5.
                                   6-3

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6.3  EVALUATION OF THE BENTHIC COMMUNITY

In a survey of the benthic community of Scippo Creek,  conducted in July
1971, effects to the community were found to extend approximately 1.6  km
downstream from the outfall (Battelle Laboratories 1971).   The present
trichopteran- and ephemeropteran-dominated community was absent from
riffle habitats according to Battelle.   Battelle Laboratories (1971)
reported an abrupt recovery of the community at a distance located
approximately 3.3 km downstream of the outfall.  However,  no collections
were made between the 1 .6- and 3.3-km sites to ascertain more specifi-
cally where recovery occurred.  They also reported that the benthic com-
munities below the recovery zone were more stable than Station 1  because
of the greater diversity values in the recovery zone.

Results of this August 1982 study revealed an improvement  in the benthic
community, as measured by the increase  in numbers of individuals and
taxa, at all sites compared to Battelle Laboratories (1971) results.
The variety of taxa and  community abundance in 1982 increased substan-
tially from 1971  indicating the benthos had a more complex community
structure.  Although mayflies and caddisflies remain major components
of the community,  by 1982 midges became numerically dominant and the
most diverse group.   In addition,  oligochaetes,  crustaceans other than
crayfish, and miscellaneous organisms were collected in 1982.

Riffle areas immediately downstream from the outfall (Station 2,  92.3-m
distance) were not devoid of biota as reported by Battelle Laboratories
(1971).  The greatest abundance in this study was found at Station 2.
However, the benthic community at Station 2 had low diversity values
compared to other stations and had a predominance of a relatively few
midge taxa.   Although habitat characteristics were similar between sta-
tions,  the flow regime differed.   In addition,  a fungal growth appeared
all over the substrate,  which also may  account for population differ-
ences.   The hydropsychids at Station 2  responded adversely to either
water quality conditions or fungal growth (Figure 6-5).
                                   6-4

-------
For the most part,  the hydropsychids (caddisflies)  are collectors and
gatherers,  whereas  the dominant midges at Station 2 are herbivores,
thus eliminating competition for food as  a factor regulating abundance.
Species of Baetis (mayflies) are herbivores and  detrital  feeders  (Merritt
and Cummins 1978) and might be considered competitive  for food with the
midges found at Station 2.   However,  the  chlorophyll a content of the
periphyton was high (Chapter 5), indicating that food  availability was
not influencing the distribution of Baetis.  Grazing pressure from the
large numbers of minnows,  particularly creek chubs  (Chapter 7), at
Station 2 was also  evaluated as a possible cause in the reduction of key
benthic taxa.  However, the total benthic population was  most abundant
at this station, suggesting that predation was not  a limiting factor to
benthic colonization.

There was a substantial increase in numbers of mayflies,  Baetis. at
Station 3 and below,  similar to the increase in  abundance of the tri-
chopterans, Cheumatopsvohe  and Hydroosvche.

Station 3 was affected by  the discharge during Battelle Laboratories'
study (1971).  In contrast,  the highest diversity value for the 1982
survey occurred at  this station, as well  as the  peak density of Baetis.
The high diversity  value and high abundance of benthic organisms depicts
a different community at Station 3 than at Station  4,  where there was
a decrease in the diversity index and a slightly different species com-
position of the benthic community.   However,  results of the x^ analysis
indicate that there was no  difference in  number  of  taxa.  The community
farthest downstream (Station 5)  had a high diversity value and the
largest number of taxa (70), but was not  significantly different from
Stations 1, 3, and  4  in number of taxa (Table D-6) .

A localized effect  on the  benthic community of Scippo  Creek was observed
at Station 2, but the conditions reported by Battelle  Laboratories (1971)
have improved.  Some  of the observed effects may be due to habitat alter-
ation by fungal growth and  deposition of  iron precipitates.  The history
of effluent treatment modifications within the 11 years between studies
was not reviewed to ascertain the reason  for the improvement.
                                   6-5

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V
>

Q
5.0-


4.0-



3.0-



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           Outfall
 o
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 70-


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 10-
                                                                     Number of Species

                                                                     Number of Individuals
            1f2

           Outfall
                                                                                          r 15,000
-10,000
         p

         3"
- 5,000
                                       Stations
              Figure 6-1.  Diversity index (d) and components of the index in Scippo Creek.
                                               6-6

-------
     6,000-
     5,000-
     4,000-
JE

 d
 c
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     3,000 -
     2,000-
     1,000 -
                   15,600
    I,

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   ;
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  /
 I
 I



V
Outfall
                                                                             Chironomidae
                                                                     	Oligochaeta
                                            Sampling Stations
           Figure 6-2.  Mean density of Chironomidae and Oligochaeta in Scippo Creek.

                      The standard deviation is indicated by brackets.
                                            6-7

-------
     2,800 -i
     2,400-
     2,000-
     1,600-
Q    1,200-
      800-
      400-
                   Trichoptera
            	Ephemeroptera
            Outfall
                                            Sampling Stations
           Figure 6-3. Mean density of Trichoptera and Ephemeroptera in Scippo Creek.
                      The standard deviation is indicated by brackets.
                                           6-8

-------
     8,800-i
     7,600-
     6,400 -
     5,200-
d
-   4,800-
c
0>
Q
     3,600-
     2,400 •
     1,200-
Chironomus spp.

Cricotopus tremulus
                                                             	Polypedilum convictum
                                           Sampling Stations
               Figure 6-4. Mean density of Chironomids (midges) in Scippo Creek.

                          The standard deviation is indicated by brackets.
                                          6-9

-------
   1,000 -I
     800-
     600-

-------
   1,400-
   1,200-
   1,000 -
-I   800-

 d
 >

 t^
'«/»

 C
     600
     400-
     200-
                                                            	  Baetis spp.


                                                            	  Early Instar Baetidae
            V
           Outfall
                                          Sampling Stations
          Figure 6-6. Mean density of Ephemeropterans (mayflies) in Scippo Creek.


                     The standard deviation is indicated by brackets.
                                          6-11

-------
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                                                                6-12

-------
TABLE 6-2  SHANNON-WIENER DIVERSITY INDICES  AND ASSOCIATED EVENNESS AND
           REDUNDANCY VALUES CALCULATED ON BENTHIC  MAC.10INVERTEBRATE
           DATA.  SCIPPO CREEK(a)
Station
1
2
3
4
5
Diversity
4.4696
2.9494
4.6697
3.8906
4.3586
(b)
Evenness
0.7630
0.5435
0.7644
0.7044
0.7111
(b)
Redundancy
0.2397
0.4572
0.2363
0.2971
0.2933
No. of
Species
58
43
69
46
70
No. of
Individuals
2,622
17,761
11,942
5,451
2,233
(a)  Calculated on a log base  2.
(b)  The sum of evenness and redundancy  pairs equals one.
                                 6-13

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                           7.   FISH COMMUNITY

The fish community is the highest trophic level  potentially  affected  by
discharges to Scippo Creek.   This survey  investigated  the fish  community
to discern any changes in composition and dominance  from previous  surveys
and to evaluate the response  at various stations.   A description of the
sampling and analytical methods is presented in  Appendix C.   Species
names and common names are provided in Appendix  D.

7.1  COMMUNITY STRUCTURE

The fish collections yielded  19 species and three  taxa of fish  that could
be identified to only the family or genus level  (Table 7-1).  Four fami-
lies were represented in the  study area,  but a maximum of three occurred
at any one station.  The stoneroller, creek chub,  sand shiner,  rainbow
darter, and Johnny darter were common species  to all five stations.
Five additional species were  encountered  upstream  at a collection  site
for resident species toxicity testing: quillback,  pumpkinseed, warmouth,
and the black and golden redhorses.

Station 1 yielded 17 species,  including seven  smallmouth bass,  one rock
bass, and one small Lepomis sp. , the only centrarchids collected (Table
7-1).  The catches at Stations 2 through  5 contained mainly  cyprinids,
with small percentages of darters and suckers.   The largest  number of
specimens was collected at Station 2.  The substantial depth  and cover in
the pool area and greater effectiveness of seining was at least partly
responsible for the larger catches.   Creek chubs and stonerollers
composed over 90 percent of the catch at  Station 2.  The numbers of
specimens and taxa caught at  Stations 3,  4, and  5  were all less than
those caught at Station 1.  The poorest species  and  family representation
occurred at Station 4, where  five species of cyprinids and four species
of darters were collected.
                                   7-1

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7.2  EVALUATION OF THE FISH COMMUNITY

A fish survey was conducted on Scippo Creek by the State of Ohio
Environmental Protection Agency (EPA) in October 1974.   The station
locations used in that study were similar to Stations 1,2, and 4 in
this study (Figure 2-1).  EPA also used 92.3-m sections,  but made 40
hauls with a 9.2 x 3.7 m deep seine at each station.

The abundance and number of species in 1982 at Station  1  were similar
to those found by the State of Ohio EPA (1974);  however,  the species
composition was somewhat different.   No darters  were collected in 1974,
whereas 25 rainbow and Johnny darters were collected in this study.
The catostomids were represented at Station 1  by a small number of fish
in 1974, but none was collected in 1982.   Also,  more centrarchids were
collected than in the previous study.

The abundance and diversity of fish found at Station 2  in 1982 far
exceeded those collected in 1974.   Four species  of darter were collected
in this study, whereas only one Greenside darter was caught in 1974.

Station 4 had the poorest family and species representation of the five
stations studied in 1982, with 235 fish from ten species and two fami-
lies.   In 1974, only 43 fish from  eight species  and three families were
collected.   The darters were well  represented in both studies,  with  four
species caught in each case.

The number of taxa collected at Stations 2 through 5  were not signifi-
cantly lower than Station 1 ,  the reference station,  as  indicated by  a
x2 test.   In contrast,  the number  of individual  fish  collected increased
400 percent from Station 1  to Station 2,  then decreased to 18-30 percent
of the catch at Station 1 for the  remaining stations (Table 7-1).   These
large differences in number of individuals were  highly  significant,  at
P 
-------
in great numbers.   Subtle variation in habitat  could account for the dif-
ferences between stations in composition and abundance within the fish
community.   In addition,  the area sampled for Station 1 was 50 percent
larger than for the other stations.

The number of species collected  at Scippo Creek varied from 10 to 17,
with the highest number collected at Station 1  (Table 7-1).  The number
of species is so similar among Stations 2 through 5 (results of a x2 test
were statistically nonsignificant) that community structure appeared to
be unchanged among the downstream stations.  The reduction in fish col-
lected downstream of Station 2 does not coincide with expected response
to effluent toxicity.  Usually toxic effects diminish downstream.  This
difference in abundance may be attributable to  either habitat differences
or enrichment of food sources.
                                  7-3

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     TABLE 7-1  ABUNDANCE OF FISH SPECIES.  SCIPFO CREEK.  AUGUST 1Q82
                                                Station
         Taxa	           1(a)     2(b)
Cyprinidae (small)                 23                         5
Cyprinid hybrid                     1
Creek chub                         90     1,469      41       50       28
Blacknose dace                      2         2817
Spotfin shiner                      8                 2               26

Bluntnose minnow                  263       131      53      117
Stoneroller                       161     1,404      21       27       22
Striped shiner                     26         2                      15
Sand shiner                        93         8       5       18       21
Silverjaw minnow                   43        56

Silver shiner                      27                22               11
White sucker                                 16       1                3
Northern hogsucker                            1                       1
Rock bass JUV                       1
Smallmouth bass YOY                 4

Smallmouth bass JUV                 3
Lepomis sp.                         1
Greenside darter                              1211
Rainbow darter                     21         4      24        5        5
Fantail darter                                2       2

Johnny darter                       4         7222
Banded darter                                         1        9
Total number of taxa(c)            17        13      13      10      12
Total number of individuals(d)    771     3,103     184     235     142
(a)  Totals from 138.5-m sampling section;  all other stations were
     92.3 m.
(b)  Aliquot procedures used.
(c)  X2 test results were:  nonsignificant  differences among stations.
     Station 1  was used as the expected value.
(d)  X2 test results were:  highly significant differences among
     stations (P £. 0.0001).  Station 1  was  used as the expected value.

Note:  JUV = juvenile.
       YOY = young of the year.
                                   7-4

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             8.   COMPARISON OF LABORATORY TOXICITY TEST DATA
                 AND RECEIVING WATER BIOLOGICAL IMPACT
One of the objectives of the Complex Effluent Testing Program is to
determine which toxicity tests best predict the receiving stream biolog-
ical impact.   Through comparative studies,  the reliability of effluent
toxicity tests for protecting the aquatic community can be determined.
Biological field surveys are useful in assessing pollutant impact,  but
are of little or no value in determining how much each discharge affects
the receiving waterbody.  In the development of permit limits,  a rela-
tionship must be established between the effluent and receiving water
impact.  Chronic toxicity tests have the potential to measure toxicity
in the receiving stream and to predict biological impact.   The major
problem in establishing this relationship is using laboratory toxicity
data from one or two species to predict the community effects for many
species.

The development of short, chronic tests has made onsite acquisition of
chronic data practical.  Toxicity data, expressed as an effect concen-
tration (e.g., the acceptable effluent concentration (AEC)),  can provide
the quantification needed to set treatment requiranents in order to
reduce toxic water quality impact.   If the AEC is not exceeded in the
stream, it can be concluded that there will be no toxic impact from the
effluent.

The AEC, as measured in the laboratory on a few species,  must compensate
for the extrapolation from toxicity data for a few tested species to an
AEC for the many species in the community.   The sensitivity of any  test
organism, relative to that of the species in the community, is not  known.
Therefore, if toxicity is found,  there is no method to predict whether
many species, or just a few, would be adversely affected at similar con-
centrations,  since the sensitivities of the species in the community also
are not known.  For example, at a given waste concentration,  if the test
species has a toxic response and if the species is very sensitive,  then
only those few species in the community of equal or greater sensitivity
would be predicted to be adversely affected.   Conversely,  if  the test
                                   8-1

-------
species is tolerant of the effluent,  then many more species in the com-
munity should be adversely affected at similar concentrations.   Thus,
the number of species lost due to a toxic effluent cannot be related to
the degree of toxicity measured in the toxicity test,  unless the position
of the tested species within the sensitivity range of  the community is
known.  In this study with only one effluent,  the position of the tested
species sensitivity would remain the same so long as the communities at
each station had the same sensitivity range.

The loss of one or two species from a community is not likely to be
considered an adverse effect.   Such small changes may  be due either to
sampling,  habitat differences, or the result of the suspected effluent.
Further, the toxicity test results only reflect toxicity over the 7-day
test period.  In contrast, the biological community is a result of adap-
tation and reaction to many past events that affected  the community which
include many factors other than the effluent.

The conceptual framework for the data comparison does  not rely on test
species being a surrogate for any one species or group of species within
any community.  The fathead minnow data are not intended to predict
only the response of the fish community,  nor are the ,C.  reticulata data
intended to predict only the response of the zooplankton community.  How-
ever, the conceptual framework does rely on the assumption that the test
species' sensitivity is within the range of the sensitivities of species
that comprise the biological community.
8.1  PREDICTIONS OF INSTREAM COMMUNITY IMPACTS BASED ON EFFLUENT
     DILUTION TEST AND AMBIENT TOXICITY TEST RESULTS
In this study, two organisms,  C_. reticulata and fathead minnows,  were
used to assess effluent toxicity.   Neither test species exhibited acute
or chronic toxic responses to the effluent.  The AEC for both species
was greater than 100 percent effluent concentration.  These results
predict no adverse effect from the discharge.   The biological survey
results revealed no conclusive evidence of toxic effect from the single
discharge in Scippo Creek.  Since species sensitivity is the basis
                                   8-2

-------
for the comparison of the toxicity tests and instream community  data,  it
is most desirable to use the total number of species/taxa collected  at
each station.  Other community measures are not regarded as  valuable as
the number of species/taxa.   The community loss index is overly  sensitive
to habitat effects.   Diversity is not useful for cases where the sensi-
tive species of the community are not dominant.

Numbers of organisms and taxa were high below the outfall,  but there was
a decrease in benthic macroinvertebrate taxa immediately below the
outfall at Station 2.  However, this decrease was probably  due to a
habitat loss, caused by the obvious clogging of the interstitial  spaces
in the substrate which the invertebrates inhabit.   If the loss of
invertebrate taxa at Station 2 were due to effluent toxicity, one would
not expect to see an increase at Station 3, a decrease at Station 4,
followed again by an increase at Station 5.  A better explanation would
be sampling variations or habitat differences.

Fish species also show a marked decrease in number of species at stations
downstream of Station 1 .  This loss may be due  to the larger area sampled
at Station 1. In addition,  the number of fish species is lower at
Stations 4 and 5 than at Stations 2 and 3.  This pattern is  not  to be
expected if effluent toxicity is the cause.

8.2  SUMMARY

The results of the Scippo Creek study demonstrated that the  tests are
practical to conduct onsite or using shipped samples.   The fungal problem
was obviously not effltent-caused, but is of concern if such tests are
to be routinely used.  Any measurement, including simple chemical ones,
occasionally fail or show interferences.   Toxicity tests are no  excep-
tion.   The fungal problem encountered in the ambient toxicity tests
(which was also observed all over the substrate in the benthic
macroinvertebrate analysis)  was conspicuous and would certainly  have
caused rejection of test results in routine uses.   The important  issue is
whether this problem occurs frequently.  Only continued use  will  tell.
                                   8-3

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The effluent toxicity tests predicted  no toxic  impact  on Scippo  Creek
from the discharge.   The field survey  found  a localized  small reduction
in the number of taxa approximately 100 m from  the  outfall  at Station 2.
This reduction is probably due to a habitat  change  from  the physical
clogging of spaces between rocks  in the stream  bed—not  from toxicity.

For regulatory use,  the correct prediction of a nontoxic effect  is  as
important as the prediction of a toxic effect.   If  the localized effect
was due to physical  alteration of the  substrate,  corrective action
imposed by a regulatory authority would be quite different  from  the case
where the localized effect was due to  toxicity.   Treatment  of the process
waste would not aid in the removal of  precipitate from the  cooling  water.
                                   8-4

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                               REFERENCES

Battelle Laboratories.  1971.  A Survey of Macrobenthic Organisms in
  Relation to Waste Water Effluents in Scippo Creek, Pickaway County,
  Ohio.   Prepared for PPG Industries, Inc.

Carter,  H. H. and A. Okubo.  1970.  Longitudal Dispersion in Non-Uniform
  Flow.   Technical Report No. 68.  Johns Hopkins Univ., Chesapeake Bay
  Institute.  45 pp.

Hamilton, M. A.  1984.  Statistical Analysis of the Seven-Day Ceriodaphnia
  reticulata Reproductivity Toxicity Test.  EPA Contract J3905NASX-1.
  16 January.   48 pp.

Merritt, R. W.  and K.  W. Cummins.   1978.  An Introduction to the Aquatic
  Insects of North America.  Kendall/Hunt Publishing Company.  441 pp.

Mount,  D.I.  and T. J.  Norberg.  1984.  A seven-day life-cycle cladoceran
  toxicity test.  Environ. Toxicol.  Chem. 3( 3):425-434.

Mount,  D.I. , ed.  In preparation.  Validity of Effluent and Ambient
  Toxicity Tests for Predicting Biological Impact in Five Mile Creek,
  Birmingham,  Alabama.  Prepared by EA Engineering, Science, and
  Technology,  Inc. and EPA Environmental Research Laboratory, Duluth
  for EPA Permits Division, Washington, D. C.   Various pagings.

Norberg, T. J.  and D.I. Mount.  In press.  A new subchronic fathead minnow
  (Pimeohales promelas) toxicity test.   Environ.  Toxicol. Chem.

Robins,  C. R. ,  R.M. Bailey, C. E. Bond, J. R. Brooker, E. A. Lachner,
  R.N.  Lea,  and W.B.  Scott, eds.   1980.  A List of Common and Scientific
  Names  of Fishes from the United States and Canada.  American Fisheries
  Society Special Publication No. 12, Fourth Edition.   Committee on Names
  of Fishes,  Bethesda, Maryland.   174 pp.
                                   R-1

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Rogers, J.  1984.  University of Wisconsin at Superior, Wisconsin, and
  EPA Environmental Research Laboratory at Duluth, Minnesota.  July.
  Personal communication.

State of Ohio.   1974.  Unpublished data.   Environmental Protection
  Agency.   7 PP.

Steele, G. R. and J.H. Torrie.   1960.  Principles and Procedures of
  Statistics,  a Biometrical Approach,  2nd edition.  McGraw-Hill,  New
  York.  633 PP.

Weber, C.I.   1973.  Recent developments in the measurement of the
  response of plankton and periphyton to changes in their environment,
  in Bioassay Techniques and Environmental Chemistry (G. E. Glass,  ed.),
  pp. 119-138.   Ann Arbor Science Publishing,  Ann Arbor,  Mich.
                                   R-2

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                A.  TOXICITY TEST AND ANALYTICAL METHODS

For the effluent dilution toxicity tests, the dilution water was col-
lected as a grab sample from just upstream (Station 1) of the outfall
during late morning of the day it was used.   The effluent was collected
as a 24-hour composite sample by continuously pumping a small quantity
from the discharge flow.  Compositing began in late afternoon and the
discharge was relatively constant.   Therefore,  the composite was
essentially flow-proportional.  Refer to Mount and Norberg (1984) and
Norberg and Mount (in press) for a detailed presentation of methods.

Onsite toxicity testing was conducted using Ceriodaphnia reticulata,
fathead minnows, and resident species.

Effluent and upstream dilution water samples were air-shipped each day
to Duluth for additional laboratory toxicity testing.   At ERL-Duluth,
the 7-day larval fathead minnow tests and the £. reticulata tests were
conducted using shipped receiving water and Lake Superior water as
diluents.

In all these tests,  new test solutions were made daily from a new 24-hour
composite effluent sample and a new grab sample of receiving water.   For
those tests using Lake Superior dilution water, a new sample was used.
The resident species were neither fed nor acclimated before the test  was
begun.   Small rocks  collected from  Scippo Creek were placed in the
benthic invertebrate test chambers  as a substrate.

For the fathead minnow and £. reticulata tests,  concentrations of 100,
25, 10, 5, and 1 percent effluent were tested.   For the resident species
tests,  only 100, 50, and 25 percent effluent concentrations were tested.

The various concentrations were made by measuring effluent and stream
water using graduated cylinders of  various sizes,  then mixing each con-
centration in a polyethylene container.   All vessels to which effluent
or ambient water was in contact were glass,  polyethylene,  or aluminum.
All samples were at  or near DO saturation when solutions were made.
                                   A-1

-------
Enough test solution was mixed in one batch for  the fathead minnow,
C. reticulataf and resident species tests.

No chemical measurements for specific chemicals  were made.   Routine  water
chemistry, such as DO and pH,  was measured  in various samples  daily.
Many of the DO measurements were made just  before changing  test  solutions
to determine the minimum values occurring.

Test solutions were changed daily so that in the effluent dilution tests,
the fish and £. reticulata were exposed to  a new 24-hour  composite
effluent sample each day which was made up  in a  new daily grab sample
of receiving water.  In addition to the effluent dilution tests,  four
ambient stations were established,  one above the outfall  and three spaced
downstream for measurement of  receiving water toxicity.   These stations
were the same as those used for the biological survey.   A daily  grab
sample was taken at each station and 10 C..  reticulata were  exposed to
each sample for 24 hours, all  in separate 30-ml  beakers  containing 15  ml
of water sample.

A.1  Ceriodaphnia TESTS

The £. reticulata were from the Duluth culture.   They were  placed one
animal to each of ten 30-ml beakers for each concentration  or  ambient
station sample tested.  Fifteen ml of test  water were placed in  each
beaker and a newly born £. reticulata. less than 6 hours  old,  was used.
One drop (0.05 ml) of a food solution containing 250 pg yeast  was added
daily.  Each day the adult was moved to a new test solution, a 15-ml
volume, with an eye dropper; food was again added.  When young were
present, they were counted and discarded.   Temperatures were maintained
at 23-25 C.  For the effluent dilution tests,  the same concentrations
were used as described for the fish.  Light was  kept very dim  to avoid
algal growth and to keep conditions comparable to those  used for
culturing at Duluth.  The culture procedures and test method are provided
in Mount and Norberg (1984).
                                   A-2

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A. 2  FATHEAD MBfNCH TESTS

For the larval fathead minnow tests,  a chamber 30.5 x 15.2 x 10.2-cm deep
was made and divided by three glass partitions which resulted in four
compartments 12.7 x 7.6 x 10.2-cm deep.   The partitions stopped 2.5 cm
short of one side of the chamber and  a piece of stainless steel screen
was glued from one chamber end to the other and across the ends of each
compartment.  This left a narrow sump 2.5 x 30.5 x 10.2-cm deep along one
side of the chamber to which each of  the four compartments was connected
by its screen end.  In this way, the  compartments could be filled and
drained by adding to or removing water from the sump but retaining the
fish in the compartments relatively undisturbed.  This design allowed
four replicates for each concentration.   These are not true replicates
in the pure statistical sense because there was a water connection
between compartments.   However,  there was virtually no water movement
between compartments as judged by DO  measurements.   (In seme cases there
were measurable DO differences between compartments.)  When the compart-
ments were filled or drained, sane water would mix into other chambers.

Each day 0.1 ml of newly hatched brine shrimp were fed three times to
the fish.  Fish survival was determined each day.  Live brine shrimp were
available during the entire daylight  period of 16 hours.  Light intensity
was low.

Each day the compartments were siphoned using a rubber "foot" on a
glass tube to remove uneaten brine shrimp.  Additional test solution was
removed from the sump until about 500 ml remained in the four compart-
ments combined, which equaled about 1 cm of depth or 10-15 percent of the
original volume.  Then, approximately 2,000 ml of new test solution were
added slowly into the sump.  The larval fish were able to easily maintain
their position against the current.  Fish were assigned to compartments
one or two at a time in sequential order.  They were less than 24 hours
post-hatch at the beginning of the test, and were obtained from the
Newtown Fish Toxicology Laboratory culture unit.
                                   A-3

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Because of inadequate temperature control  in the mobile lab,  the onsLte
tests with fathead minnows were conducted  with  temperatures varying from
18 to 25 C.  These lower temperatures reduced growth  of the minnows from
that expected at a constant 25 C.   The C.  reticulata  were kept  in a con-
stant temperature cabinet and were not so  affected.

At the end of the test,  the fish were counted and preserved in  4 percent
formalin.  Upon return to the Duluth laboratory, they were rinsed in dis-
tilled water, oven dried at 98 C for 18 hours,  and weighed to the nearest
0.01 mg on an analytical balance.   Four lots of 10 fish were  preserved at
test initiation and later weighed to give  an estimate of initial weight.
This method is described in more detail in Norberg and Mount  (in press).

A. 3  RESIDENT SPECIES TESTS

Resident species were collected from the stream above the outfall and
tested in chambers 61.0  x 15.2 x 10.2 cm arranged exactly as  the larval
fathead minnow test chambers, but each with five compartments 12.7 x
12.2 x 10.2-cm deep.   Three liters were used to fill  each chamber.  Each
day, 3 liters were added to chambers after 80 percent of the  solution was
siphoned out. Five species were tested, one species per compartment, and
two such chambers for each concentration provided duplicate  test compart-
ments for each species.   In addition, two  fish  and one crayfish species
were tested in 30.5-cm diameter battery jars filled with 10  liters of
test solution.  All but  1 liter was siphoned out each day and 10 liters
of new solution were added.  Five organisms of  each species in  each of
two replicates were used for the test.

A. 4  FISH CAGING STUDY

The caging study was conducted using commercially available  6-mm (1/4
in.) mesh metal minnow traps whose openings had been  plugged  with rubber
stoppers.  The total volume of each cage was approximately 11.5 liters.
Three cages were used at each of the four  stations and were labeled
Prep A, B, and C.  Each cage was secured to the bank  with a  light line.
                                   A-4

-------
Fish used in the caging study were collected from locations upstream
from the discharge near the Kinston Pike bridge.  The bluntnose minnow
(Pimephales notatus) was selected for its abundance and relative ease of
identification with minimal handling stress.  The fish were transported
and held in 18.9-liter buckets.

Ten fish were placed in each of three cages.  To reduce stress at each
handling, care was taken to move the fish quickly but gently in a very
fine mesh net.  Observations were made daily at approximately the same
time and the number of live fish was recorded.   Dead fish were removed
and discarded.

A.5  QUANTITATIVE ANALYSES

A.5.1  Ceriodaphnia reticulata

The statistical analyses of the £. reticulata data were performed using
the procedure of Hamilton (1984) as modified by Rogers (personal communi-
cation).  In this procedure the young production data were analyzed to
obtain the mean number of young per female per  treatment.   Daily means
were calculated and these means were summed to  derive the 7-day mean
young value.   By this method,  any young produced from females that die
during the test are included in the mean daily  estimate.   Using this
procedure,  mortalities of the original females  affect the estimate
minimally,  but the mortality of the adult is used along with the young
production to determine overall toxicity effects.   Confidence intervals
are calculated for the mean reproductivity using a standard error esti-
mate calculated by the bootstrap procedure.   The bootstrap procedure
subsamples the original data set (1,000 times)  by means of a computer
to obtain a robust estimate of standard error.

A Dunnett's two-tailed t-test  is performed with the effluent test data
to compare each treatment to the control for significant  differences.
For the ambient station data,  Tukey's Honestly  Significant Difference
Test is used  for the ambient toxicity test data to compare stations.
                                   A-5

-------
A.5.2  Fathead Minnows

The four groups'  mean weights are statistically  analyzed with  the assump-
tion that the four test chamber compartments  behave as replicates.  The
method of analysis assumes the variability  in the mean treatment response
is proportional to the number of fish  per treatment.  MINITAB  (copyright
Pennsylvania State University 1982)  was  used  to  estimate a t-statistic
for comparing the mean treatment and control  data using weighted regres-
sions with weights equal  to the number of replicates in the treatments.
The t-statistic is then compared to  the  critical t-statistic for the
standard two-tailed Dunnett's test (Steel and Torrie 1960).  The survival
data are arcsine-transformed prior to  the regression analyses  to stabi-
lize variances for percent data.
                                   A-6

-------
            B.   HYDR(LOGICAL SAMPLING  AND ANALYTICAL  METHODS

On 10 August 1982, prior to the dye release,  flow measurments were made
at Stations 1,  2, and 3 in order to assist in estimating  the arrival
time of dye at Stations 2, 3» and 4.   Additional flow measurments were
made on 10 August at Station 4 after the dye  release  and  on 13  August
at Stations 4 and 5.  The measurements were made with a Teledyne Gurley
pygmy flow-meter.  At each station,  the velocity measurements were made
along a transect with the distance between each  reading not exceeding
0.3 m and at a depth of 0.6 m of the water column.

At 1330 hours on 10 August 1982, 145.8 g of 20 percent solution of
Rhodamine WT dye was released in the effluent prior to its point of
discharge into Scippo Creek.  At Stations 2 (0.1 km),  3 (1.3 km)» and
4 (3.70 km) downstream from the point  of discharge, grab  samples were
collected near midstream at an approximate 0.1-m depth in 200-ml plastic
bottles.  The sampling interval was initially 15 minutes  at each station
and decreased as the main dye mass approached.  At Station 2, samples
were collected from 1345 to 1523 hours.   During passage of the  main dye
mass, samples were collected at 15- and 30-second intervals (1427-1438
hours).  At Station 3> samples were collected from 1600 to 1900 hours,
with a 2-minute interval used between  1653 and 1815 hours.  At  Station 4,
samples were collected from 1845 to 2245 hours with a 5-minute  interval
after 2000 hours.

Grab samples were processed in a Turner Designs fluorometer set in the
discrete sample mode.  The fluorometer had been calibrated prior to the
study and calibration was checked each day it was used with standard
dye solutions.   The fluorometer data were converted to dye concentration,
C(ppb), using the relationship:

    C(ppb) = SR exp [0.027(1-20)]                        (Equation B-1)
                                   B-1

-------
where
    S  =  slope from the calibration regression for  the
          appropriate fluorometer scale
    R  =  fluorometer reading
    T  =  temperature (C) of the grab sample at the  time
          it was processed

This relationship includes a correction  factor for the temperature
dependence of fluorescence.

Carter and Okubo (1970)  show that the dispersion  characteristics of a
channel, as measured by  the longitudinal dispersion  coefficient, may be
identified by studying the distribution  of  dye introduced as an instan-
taneous point source.  The variance  (cr2) of the longitudinal distribution
of the dye concentration, when plotted against time,  provides a relation-
ship whose slope is related to the longitudinal dispersion coefficient
(K).  Mathematically this relationship is
    K = J-Jp                                             (Equation B-2)

Carter and Okubo also show a simple method  of  calculating  the variance
by fitting a Gaussian distribution to the dye  tracer  concentration data.
The standard deviation,  square root of the  variance of  a Gaussian dis-
tribution, is given by
         1  area under concentration curve                ,_            .
         2,       peak concentration                      (Equation B-3)
The area and peak concentration parameters  of  the observed  dye  concentra-
tion data at each station may be used with  Equation B-3  in  order  to fit
an equivalent Gaussian distribution to the  data.   The resulting standard
deviation of the Gaussian distribution may  be  used with  Equation  B-2  to
calculate the longitudinal dispersion coefficient.
                                   B-2

-------
Multiplying the dispersion coefficient by  the  travel  time  (to a  point
downstream) yields an area value  that is proportional  to the distance
between the leading and trailing  edges of  the  dye  distribution multiplied
by the mean width of the river.   As a result,  the  dispersion coefficient
can be used to characterize the spatial distribution  of water particles
for a given exposure time.
                                  B-3

-------
             C.   BIOLOGICAL SAMPLING  AND ANALYTICAL  METHODS

Water quality measuranents consisting of temperature,  dissolved oxygen,
pH, and conductivity were taken at every station.  The instruments used
for water quality measurements were a Hydrolab Model 1041, a  YSI Model
57 Dissolved Oxygen Meter, and a YSI  Model  33  Salinity-Conductivity-
Temperature Meter.

C. 1  PERIPHYTON SURVEY

Natural substrates (rocks) were sampled quantitatively using  an epilithic
algal bar-clamp sampler at each of four stations (Stations  1, 2, 3, and
4).  All samples were taken from the  lower  end of riffle  areas and runs
located at each station.   Four replicate samples were taken at each
station for chlorophyll a and biomass measurements.   These samples were
filtered using 0.45-Am filters and stored in ice to  await analysis in
the laboratory.   One sample consisting of a composite of  two  bar-clamp
collections was taken from each station for cursory  identification (genus
level) and abundance estimates.  These samples were  preserved in M3 pre-
servative to await analysis.   However, identifications were not conducted
due to budget constraints.

Biomass measurements of ash-free dry  weights (AFDW)  and chlorophyll .a
were analyzed from the filters in the laboratory.  A small plug (of equal
size) was removed from each filter for chlorophyll a analyses.  Chloro-
phyll j. was determined spectrophotometrically  after  instrument calibra-
tion with a chlorophyll a standard (Sigma chemicals) extracted in a 90
percent acetone solution.  The plugs  of the filters  were  macerated, and
chlorophyll a was extracted with a 90 percent  acetone solution.  For
AFDW, the remaining portions of the filters were dried at 105 C to a
constant weight and ashed at 500 C.   Distilled water then was added
to replace the water of hydration lost from clay and other minerals.
Samples were redried at 105 C.
                                   C-1

-------
The chlorophyll a and biomass replicate  data for each station were
analyzed quantitatively by  using one-way analysis of variance (ANOVA).
In both cases,  ANOVAs were  conducted on  data from all stations and again
on data from only Stations  1,2,  and 4.   Because of the high variation in
the data, Station 3 was omitted  from the second analysis.

C.2  BENTHIC MACROINVERTEBRATE SURVEY

Benthic samples were collected from  the  pool and riffle habitats at all
five stations.   Five replicate samples were collected from each of the
two habitats at each station.  A Hess sampler  (881 cm2) was used to
sample the benthos in the pool habitat.   Because of shallow depth (5-10
cm) of the riffle habitat,  a Surber  sampler (881 cm2) was used to collect
the benthos from this habitat at each station.  The mesh size on the Hess
sampler is 363 Mmi whereas  that  of the Surber  sampler is 500 ^tm.  Samples
were preserved in 10 percent buffered formalin and returned to the labo-
ratory for analysis.  Samples from the pool habitat were not processed,
primarily due to budget constraints.  Emphasis on the riffle habitat
was believed sufficient to  detect effects.

The benthic samples contained large  amounts of detritus and organisms
and were subsampled to expedite  organism sorting and identification.
Subs am pi ing was done using  EA's  pneumatic, rotational sample splitter
(patent pending).  Samples  were  sorted with the aid of a Wild M-5 dis-
secting microscope.  Organisms were  sorted into major taxoncmic cate-
gories and preserved in 80  percent alcohol to  await identification.
Organisms were identified to the lowest  practical taxon, using
appropriate keys and references.   Oligochaetes and chironomid larvae were
mounted on microslides prior to  identification.

A x2 test was used to test  differences in the  number of  benthic taxa
among stations.  The number of taxa  encountered at the upstream station
(Station 1) was assumed to  be an estimate of the expected number of
taxa to be found at all stations of  similar habitat.
                                   C-2

-------
A one-way ANOVA was used to test  for  differences in abundance of key taxa
among stations.   The data were natural  log-transformed to ensure a normal
distribution and equal  variances  at all stations.  A Tukey's Studentized
Range Test was performed where a  significant  station effect was obtained
from the ANOVA.   Analyses were conducted using Minitab and SAS PROC GLM.

C.3  FISH SURVEYS

Fish collections were made at all five  stations on Scippo Creek (Figure
2-1).  The sections were 92.3 m long, except  at Station  1 where a
distance of 138.5 m was used.   Each section contained pool and riffle
habitats, although in varying proportions (Table C-1).   The pools were
sampled using either a  12 or 13.8 x 3.7 m bag seine with 0.32-cm mesh.
A 10.2 x 3.7-m deep straight seine with 0.32-cm mesh was used in the
riffles employing the "kick-seine" technique.  The number of seine hauls
or kick seines varied according to the  width  and other physical charac-
teristics to ensure complete sampling of the  area within the station.

The fish data were quantitatively analyzed using the X2  test on the
number of taxa per station and the number of  specimens per station.
Data for Station 1 were used as the expected  values.
                                   C-3

-------
      TABLE C-1   HABITAT CHARACTERIZATIONS OF THE  SAMPLING  STATIONS

                                    .  Percent of Station  Area
          Station                   Pool               Riffle
           1(&)                       60                  40
           2                         80                  20
           3(b)                       75                  25
           4                         70                  30
           5                         55                  45
(a)  138.5-m long station.
(b)  Pool and riffle separated  by  73.8  m of  run.
                                  C-4

-------
                             D. BIOLOGICAL DATA
       TABLE  D-l   RANKED  ABUNDANCE LISTING OF ALL  MACROINVERTEBRATES
      	COLLECTED.  SCIPPO CREEK, AUGUST  1982
   Species Name/Life Stage

CHIRONOMUS/L.
C. (CRICOTOPUS) TREMULUS GRP.
BAETIS/N.
POLYPEDILUM (S.S.) CONVICTUM/L.
CHIRONOMIDAE/P.
HYDROPSYCHIDAE/L.
CHEUMATOPSYCHE/L.
EPHEMEROPTERA/N.
EMPIDIDAE/L.
RHEOTANYTARSUS/L.
HYDROPSYCHE/L.
SIMULIIDAE/L.
TANYTARSUS/L.
THIENEMANNIMYIA GRP.
P. (PHAENOPSECTRA)/L.
C. (CRICOTOPUS) BICINCTUS GRP.
DICROTENDIPES/L.
POLYPEDILUM FALLAX GRP./L.
MICROTENDIPES/L.
CAENIS/N.
BOTHRIONEURUM VEJDOVSKYANUM
HYDROPTILA/L.
RHEOCRICOTOPUS/L.
POLYPEDILUM (S.S.) SCALAENUM/L.
HYDROPSYCHIDAE/P.
ACARINA
POLYPEDILIUM ILLINOENSE/L.
IMM TUBIF WITH CAP CHAET
DIPTERA/P.
NAIS VARIABILIS
PHYSELLA
TRICLADIDA
CRICOTOPUS TRIFASCIA/L.
RHEOTANYTARSUS/P.
TRICORYTHODES/N.
PARATANYTARSUS/L.
CLADOTANYTARSUS/L.

Number
1,599.176
846.596
574.040
362.504
347.588
341.712
310.976
286.568
280 .240
278.432
254.928
170.856
166.788
151.872
133.340
116.164
106.672
91.304
85.428
80.456
78.648
72.320
67.348
64.636
62.828
58.308
56.500
55.144
54.240
50.624
45.200
43.392
39.776
37.516
35.256
30.284
30.284

Percent
19.985
10.580
7.174
4.530
4.344
4.270
3.886
3.581
3.502
3.480
3.186
2.135
2.084
1.898
1.666
1.452
1.333
1.141
1.068
1.005
0.983
0.904
0.842
0.808
0.785
0.729
0.706
0.689
0.678
0.633
0.565
0.542
0.497
0.469
0.441
0.378
0.378
Cumulative
Percent
19.985
30.565
37.739
42.270
46.614
50.884
54.770
58.352
61.854
65.334
68.519
70.655
72.739
74.637
76.303
77.755
79.088
80.229
81.297
82.302
83.285
84.189
85.031
85.839
86.624
87.352
88.059
88.748
89.426
90.058
90.623
91.165
91.662
92.131
92.572
92.950
93.329
Note:  N. = Nymph
       L. = Larvae
       P. = Pupae
       U. = Unidentified
       S.S. = sensu strictu

       Capitalization of taxa is due to computerized format.

                                   D-l

-------
TABLE D-l (CONT.)

Species Name/Life Stage
STENELMIS/L.
MICROTENDIPES PEDELLUS/L.
RHYACODRILUS
ENCHYTRAEIDAE
IMM TUB IF W/0 CAP CHAET
HYDROPTILIDAE/P.
CERATOPOGONIDAE/L.
EMPIDIDAE/P.
ELMIDAE/L.
GASTROPODA
PROCLADIUS/L.
HYDRA
CRYPTOCHIRONOMUS/L.
SIMULIIDAE/P.
CHIMARRA/L.
ANCYLIDAE
TANYPODINAE/L.
EUKIEFFERIELLA/L.
HEPTAGENIIDAE/N.
TRICHOPTERA/P .
STENONEMA/N.
ELMIDAE/A.
PARAMETRIOCNEMUS/L.
OCHROTRICHIA/L.
CRICOTOPUS/L.
TH IENEMANNIELLA/ L .
NAIS BRETSCHERI
TRICHOPTERA/L.
PARALAUTERBORNIELLA/L .
C. (CRICOTOPUS) CYLINDRACUS GRP./L.
CHIRONOMINI/L.
NAIS PARDAHS
DUBIRAPHIA/L.
C. (CRICOTOPUS) FESTIVALIS GRP./L.
POLYPE OIL IUM OPHIODES/L.
PRISTINA L. LONGISETA
ABLABESMYIA/L.
LABRUNDINIA/L.
COLLEMBOLA U.
LIMNOPHILA/L.
PARAPHAENOCLADIUS/L .
HEXATOMA/L.
CRYPTOTENDIPES/L.
NANOCLADIUS/L.
POLYPE DILUM (P) TRIP./L.
PSEPHENUS/L.
ISOCHAETIDES CURVISETOSUS
TANYTARSUS/P.

Number
28.928
26.668
23.504
23.052
22.600
22.600
20.792
20.792
18.080
16.724
14.012
13.108
13.108
12.656
12.656
11.300
10.848
10.396
9.944
9.944
9.040
8.588
8.588
8.136
8.136
8.136
7.232
6.780
6.328
5.876
5.876
5.424
5.424
5.424
5.424
4.068
4.068
4.068
3.616
3.616
3.616
3.164
3.164
2.712
2.712
2.260
2.260
2.260

Percent
0.362
0.333
0.294
0.288
0.282
0.282
0.260
0.260
0.226
0.209
0.175
0.164
0.164
0.158
0.158
0.141
0.136
0.130
0.124
0.124
0.113
0.107
0.107
0.102
0.102
0.102
0.090
0.085
0.079
0.073
0.073
0.068
0.068
0.068
0.068
0.051
0.051
0.051
0.045
0.045
0.045
0.040
0.040
0.034
0.034
0.028
0.028
0.028
Cumulative
Percent
93.690
94.024
94.317
94.605
94.888
95.170
95.430
95.690
95.916
96.125
96.300
96.464
96.628
96.786
96.944
97.085
97.221
97.351
97.475
97.599
97.712
97.820
97.927
98.029
98.130
98.232
98.322
98.407
98.486
98.560
98.633
98.701
98.769
98.836
98.904
98.955
99.006
99.057
99.102
99.147
99.192
99.232
99.271
99.305
99.339
99.367
99.396
99.424
        D-2

-------
TABLE D-l (CONT.)

Species Name/Life Stage
ZAVRELIA GRP./L.
MICROPSECTRA/L.
FOSSARIA
CHAETOGASTER DIAPHANUS
PRISTINA LONGISETA LEIDYI
STENACRON/N.
HYDROPHILIDAE/L.
CHIRONOMIDAE/L
LARSIA/L.
CRICOTOPUS (ISOCLADIUS)/L.
CRICOTOPUS SYVLESTRIS GRP./L.
RHABDOCOELA
AULODRILUS PIGUETI
LIMNODRILUS HOFFMEISTERI
PRISTINA LONGISOMA
ISONYCHIA/N.
GERRIDAE/N.
DYTISCIDAE/L.
C. (ISOCLADIUS) LARICOMALIS GRP./L.
GLYPTOTENDIPES/L.
TANYTARSINI/L.
WAPSA MOBIL IS
ORCONECTES S. SANBORNI
ANTOCHA/L.
ORTHOCLADI INAE / L .
C. (CHIRONOMUS) THUMMI (RIPARIUS) GRP./L.
PSEUDOCHIRONOMUS/L.
STICTOCHIRONOMUS/L.
PLEUROCERIDAE
AULODRILUS LIMNOBIUS
LIMNODRILUS CERVIX
PRISTINA BREVISETA
ASTACIDAE
HEXAGENIA/N.
HYDROPTILIDAE/L.
BRACHYCENTRIDAE/L .
LEPTOCERIDAE/L.
TANYPUS/L.
PARATENDIPES/L.
POLYPEDILUM SIMULANS/L.

Number
2.260
2.260
1.808
1.808
1.808
1.808
1.808
1.808
1.808
1.808
1.808
1.356
1.356
1.356
1.356
1.356
1.356
1.356
1.356
1.356
1.356
0.904
0.904
0.904
0.904
0.904
0.904
0.904
0.452
0.452
0.452
0.452
0.452
0.452
0.452
0.452
0.452
0.452
0.452
0.452

Percent
0.028
0.028
0.023
0.023
0.023
0.023
0.023
0.023
0.023
0.023
0.023
0.017
0.017
0.017
0.017
0.017
0.017
0.017
0.017
0.017
0.017
0.011
0.011
0.011
0.011
0.011
0.011
0.011
0.006
0.006
0.006
0.006
0.006
0.006
0.006
0.006
0.006
0.006
0.006
0.006
Cumulative
Percent
99.452
99.480
99.503
99.525
99.548
99.571
99.593
99.616
99.638
99.661
99.684
99.701
99.718
99.735
99.751
99.768
99.785
99.802
99.819
99.836
99.853
99.864
99.876
99.887
99.898
99.910
99.921
99.932
99.938
99.944
99.949
99.955
99.960
99.966
99.972
99.977
99.983
99.989
99.994
100.000
       D-3

-------
   TABLE  D-2   NUMBER OF INDIVIDUALS  AND  PERCENT  COMPOSITION FOR BENTHIC
Station 1
Replicate 1

Species. Lifestaite
CHIRONO.-IUS, L
C. (CRICOTOPU5) TREMULUS
BAETIS, N
POLYPEDILUM (S. S. ) CONVI
CHIRONOMIDAE, P.
HYDROPSYCHIDAE. L.
CHEU.'IATOPSYCHe, L.
EPH-NFROPTERA. N.
EHPIDIDAE, L.
RHEOTAMYTARSU3 L.
HYDKOPSYCHE, L.
SlhULJIDAE, L.
TANYTrtRSUS L.
THItNEMANNIMYlA, GRP.
P. (PH^ENOPSECTRA) L.
C. (CRICOTOPUS) BICINCTU
DICROTtNDIPES L.
POLYPEDILUM FALLAX GRP.
MICROTEMDIPES, L.
CAEHIS. N.
DOTHRIONEURUM VEJDOVSKVA
HYDROPTILA, L.
RHECCRICOTOPU3. L
POLYPEDILUM (S S. ) SCALA
HYDROPSYCHIDAE. P.
AC AR I IMA
POLYPEDILIUM ILLINOENSE,
IMM TUB IF WITH CAP CHAET
DIPTERA P.
NAIS VARIABILIS
PHYSELLA
TRICLADIDA
CRICOTCPUS TRIFASCIA. L.
RIIEOTANYTARSU5. P
TRICORYTHODES, N.
PARATANYTARSUS, L.
CLADOTANYTARSUS L.
STENELMIS L.
OTH^R SPECIES


Number
0.
0.
350
22.
33.
666.
113.
56.
169.
22.
305
0.
79.
282.
0.
45.
0.
0.
485.
0.
0.
0.
0.
0.
11.
56.
0.
0
0.
0.
0.
0.
0.
0.
36.
0.
0.
0.
259.
00
00
30
60
90
70
00
50
50
60
10
00
10
50
00
20
00
00
90
00
00
00
00
00
30
50
00
00
00
00
00
00
00
00
5O
00
00
00
90
z
Comp.
0. 00
0. 00
11. 61
0 75
1. 12
22. 10
3. 75
1. 87
5. 63
0. 75
10. 11
0. 00
2. 62
9. 36
0. 00
1. 50
0. 00
0. 00
16. 10
0. 00
0. 00
0 00
0. OO
0. 00
0. 37
1. 87
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
1. 87
0. 00
0. 00
0. 00
8. 61
Replicate 2

Number
0. 00
33 90
24B. 60
11. 30
45. 20
124. 30
56. 50
124 30
135. 60
33. 90
56. 50
0. 00
124 30
1 58. 20
0 00
45 20
0. 00
0. 00
113. 00
0. 00
0. 00
56. 50
0 00
0 00
0. 00
0 OO
0. 00
0. 00
11. 30
0. 00
M. 30
0. 00
0. 00
0. OO
33 90
0. 00
33. 90
0 00
259. 90
Z
Comp.
0. 00
1. 97
14. 47
0. 66
•2. 63
7. 24
3. 29
7. 24
7. 89
1. 97
3. 29
0. 00
7. 24
9. 21
0. 00
2. 63
0. 00
0. 00
6. 58
0. 00
0. 00
3. 29
0. 00
0. 00
0. 00
0. OO
0. OO
0. 00
0. 66
0. 00
0. 66
0. 00
0. 00
O 00
1. 97'
0. 00
1. 97
0. 00
15. 13
Replicate 3

Number
0. 00
0. 00
485. 90
124. 30
22. 60
621 50
124. 30
124. 30
135. 60
11 30
124. 30
0. 00
101. 70
180. 8O
0 00
33. 90
0. 00
0. 00
146. 90
0. OO
0. 00
259. 90
33. 90
0. 00
11. 30
11. 30
O..OO
0. 00
45. 20
11. 30
79. 10
11. 30
0. 00
0. 00
22. 60
0. 00
11. 30
0. OO
350. 30
Z
Cprop ,
0. 00
0. 00
15. 75
4. 03
0. 73
20 15
4. 03
4. 03
4. 40
0. 37
4. 03
0. 00
3. 30
5 86
0. 00
1. 10
0. 00
0. 00
4. 76
0. 00
0. 00
8. 42
1. 10
0. OO
0. 37
0 37
0. 00
0. 00
1. 47
0. 37
2. 56
0. 37
0. 00
0. OO
0. 73
0. 00
0 37
O 00
1 1 . 36


Replicate 4

Number
0. 00
22. 60
90. 40
0 00
0. 00
113. 00
22. 60
11. 30
45. 20
0. 00
11 30
0. 00
113. 00
67. 80
0. 00
22. 60
0. 00
0. OO
22. 60
22. 60
0 00
90. 40
0. CO
0. 00
0. 00
11. 30
0 00
0. 00
22. 60
0. GO
0. 00
0. 00
O. OO
0. 00
0. 00
0. OO
0 00
0. 00
180. 80
Z
Comp.
0. 00
2. 60
10. 39
O. 00
0. OO
12. 99
2. 60
1. 30
5. 19
0. 00
1. 30
0. 00
12. 99
7. 79
0. 00
2 60
0. 00
0. 00
2. 60
2. 60
0. 00
10. 39
0. 00
0 00
0. OO
1. 30
0 00
0. 00
2. 60
0. 00
0. 00
0. 00
0. OO
0. OO
O. OO
0. 00
0. 00
O. 00
20. 78


Replicate 5

Number
22. 60
56. 50
192. 10
0. 00
79. 10
350. 30
56. 50
11. 30
67. 6O
0. 00
22 60
0. 00
994. 40
271. 20
0. 00
56. 5O
372. 90
0. 00
508. 50
214. 70
O. 00
226. 00
0. OO
0. 00
33. 90
O. OO
0. 00
11. 30
11. 30
0 00
0. 00
0. 00
0. 00
0. 00
101. 70
0. 00
214. 70
45. 2O
497. 20
Z
Comp.
0 51
I 28
4. 35
0. 00
1. 79
7. 93
1 28
0. 26
1. 53
0. 00
0. 51
0. 00
22 51
6. 14
0. 00
1. 28
8. 44
0. 00
11. 51
4. 86
0. 00
5 12
0. 00
0. 00
0. 77
0. OO
0. 00
0. 26
0. 26
0 00
0. 00
0. 00
0 OO
0. OO
2. 30
0. OO
4. 86
1. 02
11. 25
               TOTAL
                         3017. 10
                                        1717. 60
                                                      3084 90
                                                                      870. 10
                                                                                   4418. 30
(a)  S.S. - Sensu strictu
     L. " Larvae
     P. - Pupae
     N. * Nymph

Note: Abbreviations and capitalization of  species names are due to computer format.
                                              D-4

-------
                    TABLE D-2 (CONT.)



Replicate 1

Species. Lifestaee
CHIRONOMUS, L.
C.  TREMULUS
BAETIS, N.
POLYPEDILUtt  L.
C. (CRICOTOPU5) BICINCTU
DICR07ENDIPES L.
POLYPEDILUM FALLAX GRP.
MICROTtNDIPES, L.
CAEN IS, N.
BOTHRICNEURUM VEJDOVSKYA
HYDROPTILA. L.
RHECCRICOTOPUS. L.
POLYPEDILUM (S. S. ) SCALA
HYDROPSYCHIDAE, P.
ACARIMA
POLYPEUILIUM ILLINOENSE.
IMM TUB IF WITH CAP CHAET
DIPTERA P.
NAIS VARIABILIS
PHYSELLA
TRICLADIDA
CRICOTOPUS TRIFA5CIA, L.
RHEOFANYTARSUS. P.
TRICORYTHODES, N.
PARATANYTARSUS, L.
CLADOTANYTARSUS L.
STEMELMIS L.
OTWR SPECIES

Number
4936. 80
6768 70
226 00
418 10
1039 60
0. OO
0. 00
0. 00
226. 00
0. 00
0. OO
0.00
135. 60
565 OO
135. 60
0. 00
983. 10
135. 60
0. OO
361. 60
768. 40
45.20
0. 00
135. 60
0. 00
0 OO
0 OO
90. 40
0 OO
90 40
723. 20
0 00
0 00
0. OO
0. 00
0. 00
0. OO
O. OO
1 1 3O. OO
z
Comp.
26 06
35. CO
1. 20
2 21
3 50
0. 00
0. 00
0. 00
1. 20
0. 00
0. 00
0. 00
0. 72
2. 99
0. 72
0. 00
5. 20
0. 72
0. 00
1. 91
4. 06
0 24
0. 00
0. 72
0. 00
0. 00
0. 00
0. 48
O. 00
0. 40
3. 33
0. 00
0. OO
0. 00
0. OO
0. 00
0. 00
0 00
5. 98


Replicate 2

Number
13469. 60
7438. 00
226. 00
288 1 50
632. 80
0. 00
0.00
90. 40
45.20
0.00
0. 00
0.00
0. 00
0. 00
237. 30
237. 30
485. 90
1197. 80
0 00
203. 40
406. 80
0.00
0. 00
237. 30
0. 00
0. 00
0. 00
90. 40
0 00
0. 00
0. 00
45. 20
0. OO
0. 00
0. 00
0. 00
0.00
45. 20
406. 80
I
Conip.
47 43
26. 26
0 80
10. 15
2 23
0 00
0. 00
0 32
0. 16
0. 00
0. 00
0. 00
0. 00
0 00
0 84
0 84
1. 71
4 22
0. 00
0. 72
1. 43
0. 00
0 OO
0 84
0. 00
0 OO
0. 00
0. 32
0. 00
0. 00
0. 00
0. 16
0. OO
0 00
O 00
0. 00
0 00
O 16
1 43
Station
2
Replicate 3

I
Number Comp .
8925. 30
1050. 90
0. 00
1322. 10
768. 40
0. 00
0. 00
0. OO
0. 00
0. 00
0. 00
0. 00
0. 00
135. 60
1977. 50
0. 00
259. 90
395. 50
0. 00
226. OO
361. 60
45. 20
0 00
O. OO
0. 00
0. OO
0 00
0. 00
271. 20
0. 00
45. 20
0. 00
0. 00
0. 00
O. 00
0. 00
0. 00
45. 20
452. OO
54. 54
6. 49
0. 00
8. 17
4. 75
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
0. 84
12. 22
0. 00
1. 61
2. 44
0. OO
1. 40
3. 23
0. 28
0. 00
0. 00
0. 00
0. OO
0. 00
0. 00
1. 68
0. 00
0. 28
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
0. 28
2 79



Replicate 4

J

Number Corap.
5265. 80
1276. 90
70. 40
1197. 80
587. 60
0. 00
0. 00
135. 60
0. 00
0. 00
0. 00
0. 00
0. 00
395. 50
237. 30
79. 10
79. 10
0. 00
0. 00
406. 80
135. 60
0. 00
79. 10
1 56. 20
0. 00
45. 20
0. 00
0. 00
*5. 20
0. 00
135. 60
0. 00
0 00
0. 00
0. 00
79 10
0 00
90 40
271. 20
48.
11.
0.
11.
5.
0.
0.
1.
0.
0
0.
0.
0.
3.
2.
0.
0.
0.
0.
3.
1
0.
0.
1.
0.
0.
0.
0.
0
0.
1.
0.
0
0
0
0.
0.
0.
2.
80
83
84
1O
45
00
00
26
00
OO
OO
00
OO
66
2O
73
73
00
00
77
26
00
73
47
00
42
00
00
42
00
26
00
00
00
00
73
00
84
51


Replicate 5

J
Number Comp .
7345. OO
1491. 60
45 20
689. 30
406. BO
0. 00
0. 00
135. 60
90 40
0. 00
0. 00
0. 00
0. 00
226. OO
689. 30
1 13. 00
339. 00
1 13. 00
0. 00
271. 20
271 20
0. OO
0. OO
226. 00
45. 20
0. OO
0. 00
1130. 00
0 00
0. 00
45. 20
0. 00
0. 00
0. 00
0. 00
O OO
0. 00
271 20
587. 60
50. 54
1O. 26
0. 31
4. 74
2. SO
0. 00
0. 00
0. 73
0. 62
0. 00
O. OO
0. 00
0. 00
1. 56
4. 74
0. 78
2. 33
O. 78
0. 00
1. 87
1 87
O. 00
0. 00
1. 56
0. 31
0. 00
0. 00
7. 78
0. 00
0. OO
0. 31
0 00
0. 00
O.-OO
0. OO
0. 00
0. 00
1. 87
4. 04
TOTAL
         18904. 9O
                      28396. 90
                                      16181. 6O
                                                   10791. 50
                                                                 14531  80
                             D-5

-------
                     TABLE  D-



Replicate 1
(a)
Species. Lifestaee
CHIRONOi-IUS, L.
C. (CRICOTOPUS) TREMULUS
BAETIS, N.
POLYPEDILUM (S S > CQNVI
CHIRONOIIDAE, P.
HYDROPSYCHIDAE, L.
CHEUMATOPSYCHE, L.
EPH^MEROPTERA, N.
EMPIDIDAE, L.
RHEOTAUYTARSUS L.
HYDROPSYCHE. L.
SIMUuIIDAE, L.
TANYT^RSUS L.
THIfeNEMANNINYIA. GRP
P. (PHAENOPSECTRA) L.
C. (CRICOTOPUS) BICINCTU
DICROTENDIPES L.
POLYPEDILUM FALLAX GRP.
MICROTtNDIPES, L
CAENIS. N
BOTHRIGNEURUM VEJDOVSKYA
HYDROPTILA, L.
P.HECCRJCOTOPU5, L.
POLYPEDILUM (S. S. ) SCALA
HYDROPSYCHIDAE, P.
ACARIN'A
POLYPEDILIUM ILLINOENSE.
I MM TUB IF WITH CAP CHAET
DIPTERA P
NAIS VARIABILZS
PHYSEXLA
TRICLADIDA
CRICOTOPUS TRIFASCIA. L.
RHEOTANYTARSUS. P.
TRICQRYTHODES. N.
PARATANYTARSU3. L.
CLADOTANV TARSUS L.
STENELMI5 L.
OTHER SPECIES

Z
Ninnbcr Coop *
90 40
308. 5O
1118. 7O
452. 00
632. 80
0. 00
214. 70
0 00
723. 20
599. 90
418. 10
214. 70
305. 10
180. 80
56. 50
305. 10
33. 90
56. 50
146. 90
33. 90
0 00
33. 90
180. 60
90. 40
361. 60
124. 30
0. 00
0. 00
0. 00
146. 90
33. 90
146. 90
180. 80
0. 00
0. OO
124. 30
0. OO
33. 90
429 40
1. 13
6. 37
14. 02
5. 67
7. 93
0. 00
2. 69
0. GO
9. 07
7. 51
5. 24
2. 69
3. 82
2. 27
0. 71
3. 82
0. 42
0. 71
1. 84
O. 42
0. 00
0. 42
2. 27
1. 13
4. 53
1. 56
0. 00
0. 00
0. 00
1. 04
0. 42
1. 84
2. 27
0. 00
0. 00
1. 56
0. 00
0. 42
5. 38



Replicate 2

Numbe
0.
474.
1231.
655.
1118.
0.
214.
485.
1175.
1413.
395.
813.
429.
124.
O.
689.
0.
9O.
124.
33.
0.
18O.
214.
0.
474.
18O.
0.
0.
146.
214.
0.
180.
0.
0.
0
90.
O.
0.
734.

Z
_r Coiap .
00
60
70
40
70
00
70
90
20
50
50
60
40
30
OO
30
00
40
30
9O
00
80
70
00
60
BO
00
00
90
70
00
80
00
00
00
40
OO
00
50
0. 00
3. 99
10. 36
5. 51
9. 41
0. 00
1. 81
4. 09
9. 89
11. 88
3. 33
6. 84
3. 61
1. 05
0. 00
S. 80
0. 00
0. 76
1. 05
0. 29
0. 00
1. 52
1. 81
0. 00
3. 99
1. 52
0. 00
0. 00
1. 24
1. 81
0. 00
I. 52
0 00
0. 00
0. 00
0, 76
0. 00
0. 00
6. 18
Station
3
Replicate 3

Z
Number Cotnp .
0 00
858 60
2079 2O
632. 8O
678 00
361 60
632 8O
904 OO
1943 60
1039 60
723. 20
2305 2O
452 00
361. 60
O OO
587 60
90 40
ISO 80
45. 2O
45. 20
0. 00
271. 20
226 OO
0 OO
406 90
226 00
0 OO
0 00
90 4O
316. 40
45. 2O
90 40
316 40
0 OO
0. OO
226. 00
0. 00
0 00
813 60
0 00
5. 07
12. 27
3. 73
4. 00
3. 13
3. 73
5. 33
11. 47
6. 13
4. 37
13. 60
2 67
2. 13
0. 00
3 47
0. 33
1. 07
0. 27
0. 37
0 00
1 60
1. 33
0. OO
2. 40
1. 33
0. 00
0 00
0 53
1. 87
0. 37
0. 53
1 37
0 00
0 00
1. 33
0 00
0. 00
4 SO


Replicate 4

Z
Number ,CoraD .
0. 00
936. 20
632 80
553. 70
971. 80
632. 80
271. 20
958. 80
700. 60
0 00
339. 00
282. 50
553 70
113. 00
0. 00
440. 70
0. 00
1 13. 00
271. 20
22. 60
0 00
226. 00
497 20
0 00
22. 60
134. 30
0. OO
0. 00
305. 10
150 20
0. OO
33. 90
497 20
9-37. 90
11.30
226. 00
113. 00
0 00
576. 30
0 00
7 39
5. 59
4 89
8. 58
5 59
2 40
7. 58
6. 19
O. 00
2 99
2 50
4. 89
1 00
0. 00
3 89
0. 00
1. 00
2. 40
0 20
0. 00
2. 00
4. 39
0 00
0 20
1 10
O. OO
0 00
2 69
1 40
0 00
0. 30
4. 39
8. 28
0. 10
2 OO
1. 00
0. 00
5 09


Replicate 5

Z
Number Como.
0 00
271. 20
2260. 00
0. 00
632. 80
768. 4O
316. 40
632. 80
361. 60
723. 20
497 20
587. 60
632. 80
226. 00
0 00
226. 00
0. 00
0. OO
0. 00
0. 00
0 00
9O. 40
316. 40
0. 00
45. 20
226. 00
497. 20
0. 00
271. 20
316. 40
0 00
90. 40
0. 00
0. 00
0. 00
0. OO
0. 00
45. 20
1536. 80
- — •- -'
0 00
2 34
19. 53
0. 00
5. 47
6. 64
2. 73
5. 47
3. 12
6. 25
4. 30
5. 08
5. 47
1. 95
0. 00
1. 95
0. 00
0. OO
0. 00
0. OO
O. 00
0. 78
2. 73
0. 00
0. 39
1. 95
4. 3O
0. 00
2- 34
2.73
0. 00
0. 78
O. 00
0. 00
0. 00
0. 00
0. 00
0. 39
13. 28
TOTAL
          7977. 80
                       11887 60
                                      16950. 00
                                                        S. 60
                                                                     11571. 20
                               D-6

-------
                     TABLE  D-2 (CONT.)




Replicate 1
(a)
Species. Lifestage
CHIRONOMUS, L.
C. 
-------
                   TABLE D-2  (CONT.)



Replicate 1

Speciei. Lifescaze
CHIRONCJNUS, L.
C. (CRICOTOPU3) TREMULUS
BAETIS. N
POLYPEDILUM (S S. > CONVI
CHIRONOHIDAE, P
HYDROPSYCHIDAE. L.
CHEUMATOPSYCHE. L.
EPHEMPROPTERA, N.
EMPIDIDAE, 1_.
RHEOTANYTARSUS L
HYDROPSYCHE, L.
si nut 1 1 DAE. u.
T ANY TARSUS L.
THIPNEMANNIMYIA. GRP
P (PHAENOPSECTRA) L.
C. (CRICOTOPUS) BICINCTU
D1CRQTENDIPES L.
POLYPEDILUM FALLAX GRP
MICROTtNDIPES. L.
CAENIS. N.
DOTHRICNEURUM VEJDOVEKYA
HYDROPTILA, L.
RHECCRICOTOPUS, L.
POLYPEDIUUM (S. S. ) SCALA
HYDROPSYCHIDAE, P.
ACARINA
POLYPEDILIUM ILLINOENSE,
IMM TUG IF WITH CAP CHAET
DIPTERA P
MAIS VARIABILIS
PHYSELLA
TR I CLAD I DA
CRICOTCPUS TRIFASCIA, L.
RHEOTANYTARSUS, P.
TRICORYTHODES, N.
PARATANYTARSUS, L.
CLADOTANYTARSUS L.
STENELMIS L.
OTH«=R SPECIES

Number
0 GO
0 00
745. SO
67. BO
90. 40
67 80
90. 40
237 30
45. 20
US 60
90. 40
0.00
11.30
33. 90
0.00
0.00
0.00
0. 00
56. 50
U. 30
0 00
0.00
22.60
293. ao
11.30
33. 90
0. 00
11.30
0.00
0. 00
11.30
0. 00
0. 00
0.00
45.20
0. 00
22. 60
0. 00
124.30
Z

[

Conp.
0.
0
33.
3
4
3
4
10.
i
6.
4.
0.
0.
1.
0.
0.
0
0
2.
0.
0
0.
1.
13
0
1.
0.
0.
0.
0.
0.
0
0.
0.
2.
0.
1.
0
5.
00
00
00
00
00
00
00
50
00
00
00
00
50
50
00
00
00
00
50
50
00
00
00
00
50
50
00
50
00
00
50
00
00
00
00
00
00
00
50


Rep Lie at; 2

Ifumber
33. 70
0. 00
0. 00
0. 00
33. 90
0. 00
0. 00
33. 9O
0. 00
O. OO
0. 00
O. OO
11. 30
0. 00
0. 00
0. 00
22. 60
0. 00
0. 00
124. 30
0. 00
U. 30
0. 00
429. 40
0. 00
0. 00
0. 00
1 1. 3O
11. 30
11. 30
0. 00
0. 00
0. 00
0. 00
0. 00
0. OO
305. 10
0. 00
994. 40
z
Coap.
1. 67
0 OO
0. 00
0. 00
1. 67
0 00
0. 00
1. 67
0. OO
0. 00
0. 00
0. 00
0. 56
0. 00
0. 00
0. 00
1. 11
0. 00
0. 00
6. 11
0. 00
0. 56
0. OO
21. U
0. 00
0. 00
0. 00
0. 56
0. 56
0. 56
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
IS. 00
0. 00
48. 89
Station
5
Replicate 3

Sugber
0. 00
0. 00
361. 6O
0 OO
22. 60
77 10
67 80
79 10
23. 6O
0. CO
67 80
0 00
11 30
11. 3O
0. OO
0. OO
0 00
0 00
45. 2O
0. 00
11. 30
0. 00
11. 30
11. 30
0. 00
0. 00
0 00
0. 00
0. 00
0 00
0 00
0 OO
0 OO
0 OO
?2 60
0. OO
0. 00
U. 30
56. 50
:
Comp.
0 00
0. OO
40. 51
0. 00
2. 53
S 86
7. 59
8 86
3. 53
0. 00
7. 5"
0. OO
1. 27
1. 27
0. 00
0. 00
0. 00
0. 00
5. 06
0. 00
1 27
0 00
1. 27
1 27
0. 00
0. 00
0. OO
0. 00
0 00
0 00
0 00
0 00
0. 00
0. 00
2. 53
0. 00
0. 00
1. 27
6. 33


Replicate 4

ffomber
0. 00
11. 30
429. 40
0. 00
79 10
237 30
519 80
949 20
45. 20
67 80
372. 90
11. 30
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
0 00
0 00
11. 30
22 60
0. 00
0. 00
0. OO
22. 60
0. OO
11. 30
0 00
0. 00
0. 00
0. OO
0. 00
90 4O
0. 00
11. 30
33. 9O
429 40
Z
Comp.
0. 00
0. 34
12. 79
0. 00
2. 36
7 07
15. 49
28. 28
1. 35
2. 02
11 11
0. 34
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
0. 34
0. 67
0 00
0 00
0. 00
0. 67
0. 00
0. 34
0. 00
0. 00
0. OO
0. 00
0. 00
2. 69
0. 00
0. 34
1. 01
12. 79


Replicate 5

Number
0. 00
45. 20
237. 30
33. 9O
79. 10
192. 1O
203. 40
824. 90
67. 80
90. 40
293. 80
0. OO
22. 60
0. 00
0. OO
0. 00
0.00
0.00
169. 50
11. 30
0 00
0. 00
0. 00
33. 90
22. 60
22. 60
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
0. 00
67 80
11. 30
11. 30
22. 6O
1 58. 20
Z
Comp^
0 00
1. 72
9 05
1 29
3. 02
7 33
7 76
31. 47
2. 59
3. 45
11. 21
0. 00
0. 86
0. 00
0. OO
0 00
0. 00
0. 00
6. 47
0. 43
0. OO
0. 00
0. 00
1. 29
0. 86
0. 86
0. OO
0 00
0. OO
0. 00
0. 00
0. 00
0 00
0. 00
2. 5"
0 41
0. 43
0 86
h 03
TOTAL
         2260. 00
                       2034. 00
                                      892. 70
                                                   3356 10
                                                                 2621. 60
                              D-8

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                TABLE  D-2 (CONT.)
Species. Lifestaee "'


CHIRONOMUS, L.
C. (CRICOTOPU3) TREMULUS
BAETIS, N
POLYPEDILUM (S. S. ) CONVI
CHIRONOrllDAE, P.
HYDROPSYCHIDAE, L.
CHEUMATQPSYCHE, L.
EPHlEMFROPTERA. N.
EMPIDIDAE, L.
RHEOTANYTARSUS L.
HYDROPSYCHE, L.
SIMULIIDAE, L.
TANYTARSUS L.
THIFNEMANNIMYIA, GRP.
P. (PHAENOPSECTRA) L.
C. (CRICOTOPU3) BICINCTU
DICR07ENDIPES L.
POLYPEDILUM FALLAX CRP.
MICRCTENDIPES, L.
CAENIS, N.
DOTHRICNEURUrt VEJDOVSKYA
HYDROPTILA, L.
RHECCRICOTOPUS. L.
POUYPEDILUM (S. S. ) SCALA
HYDROPSYCHIDAE. P.
ACARINA
POLYPEDILIUM ILLINOENSE,
IMM TUB IF WITH CAP CHAET
DIPTERA P
NAIS VARIABILIS
PHYSELLA
TRICLADIDA
CRICOTCPUS TRIFASCIA, L.
RHEOTAMYTARSUS. P.
TRICORYTHODES, N.
PARATANYTARSUS, L.
CLADOTANYTARSUS L.
STENELMIS L
OTHPR SPECIES
Total

Number
1599. 18
846 60
574. 04
362. SO
347. 59
341. 71
310. 98
2S6. 57
280. 24
278. 43
254. 93
170. 86
166. 79
151. 87
133. 34
116. 16
1O6. 67
91. 30
85. 43
BO. 46
7S. 65
72. 32
67. 35
64. 64
62. 83
58. 31
56. 50
55. 14
54. 24
50. 62
45. 20
43. 39
39. 78
37. 52
35. 26
30 28
30. 28
28. 93
504. 88

Z
Comp .
19. 99
10 58
7. 17
4. 53
4. 34
4. 27
3. 89
3. 58
3. 50
3. 48
3. 19
2. 14
3 OB
1 90
1. 67
1. 45
1. 33
1. 14
1. 07
1. 01
0. 98
0. 90
0. 84
0. 81
0. 79
0. 73
0. 71
0. 69
0. 68
0. 63
0 56
0. 54
0. 50
0. 47
0. 44
0. 38
0. 38
0. 36
6. 31
TOTAL
              Q001. 75
                       D-9

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TABLE D-3  ANALYSIS OF VARIANCE AND TUKEY1 S STUDENTIZED RANGE
           TEST RESULTS FOR SPECIES OF CHIRONOMIDAE,  SCIPPO
           CREEK.  AUGUST 1Q82
Dependent Variable:
Source
Model
Error
Corrected total
Station
(mean In count)
Dependent Variable:
Source
Model
Error
Corrected total
Station
(mean In count)
Dependent Variable:
Source
Model
Error
Corrected total
Station
(mean In count)
Chironomus spp.
In count
Sum of Mean
D.f Squares Square F Value
4 157.05 39.26 111.63
20 7.03 0.35
24 164.08
Tukey's Studentized Range Test
2351
(6.49) (0.44) (0.28) (0.22)
Chironomus tremulus
In count
Sum of Mean
Df Sauares Square F Value
4 114.16 28.54 62.22
20 9.17 0.46
24 123-34
Tukey's Studentized Range Test
2315
(5.41) (3.89) (0.86) (0.46)
Polvpedilum convictum
In count
Sum of Mean
Df Squares Square F Value
4 73.97 18.49 16.63
20 22.24 1.11
24 96.20
Tukey's Studentized Range Test
2315
(4.55) (3.15) (0.86) (0.67)
PR > F
0.0001
4
(0)
PR > F
0.0001
4
(0)
PR > F
0.0001
4
(0)
                             D-10

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TABLE D-4  ANALYSIS OF VARIANCE AND TUKEY'S SIUDENTIZED RANGE
           TEST RESULTS FOR SPECIES OF HYDROPSYCHIDAE, SCIPPO
           CREEK.  AUGUST 1Q82

Dependent Variable:
Source
Model
Error
Corrected total

Station
(mean In count)
Dependent Variable:
Source
Model
Error
Corrected total

Station
(mean In count)
Dependent Variable:
Source
Model
Error
Corrected total

Station
(mean In count)
Hvdropsvche spp.
In count
Sum of Mean
Df Squares Square F Value
4 48.99 12.25 18.43
20 13.29 0.66
24 62.28
Tukey's Studentized Range Test
4351
(3.81) (3.73) (2.19) (1.88)

Cheumatopsvche spp.
In count
Sum of Mean
Df Squares Square F Value
4 55.07 13.77 26.02
20 10.58 0.53
24 65.65
Tukey's Studentized Range Test
4351
(4.43) (3-32) (2.19) (1.91)

Early Instar Hydropsvchidae
In count
Sum of Mean
Df Squares Square F Value
4 40.19 10.05 4.54
20 44.26 2.21
24 84.45
Tukey's Studentized Range Test
4135
(3.60) (3.29) (2.35) (2.00)


PR > F
0.0001

2
(0)


PR > F
0.0001

2
(0)


PR > F
0.0090

2
(0)
                             D-ll

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      TABLE D-5  ANALYSIS OF VARIANCE  AND TUKEY'S STODENTIZED  RANGE
                 TEST RESULTS PERFORMED ON SPECIES OF BAETIDAE,
     	SCIPPO CREEK.  AUGUST  1982	
                               Baetis sp.
Dependent Variable:   In count
Source
Model
Error
Corrected total
Station
(mean In count)
Dependent Variable:
Source
Model
Error
Corrected total
Sum of
Df Squares
4 23.30
20 20.40
24 43.71
Tukey's Studentize
3
(4.77) (4
In
££
4
20
24
Earlv Instar
count
Sum of
Squares
15.50
34.58
50.08
Mean
Sauare F Value
5.83 5.71
1.02
id Range Test
4 1 5
.04) (3-09) (2.89)

Baetidae
Mean
Sauare F Value
3.87 2.24
1.73
PR > F
0.0031
2
(1.98)
PR > F
0.1009
Tukey's Studentized Range Test not performed  since  ANOVA results were
nonsignificant.
                                   D-12

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   TABLE D-6  RESULTS OF A x2 TEST PERFORMED ON  THE  NUMBER  OF BENTHIC
   	MACROINVERTEBRATE TAXA COLLECTED AT EACH  STATION	
                                                 Station
Number of taxa(a)                 53       43        59       46       70

Expected number
  (based on Station 1)             —       58        58       58       58

X2 contribution(b)                 —     3.88(c)   2.09     2.48     2.48
(a)  Number of unique taxa/life  stages  by  combining five replicate
     samples for each station.
(b)  For individual  stations,  the  1  degree of freedom x2 with
     P > X2 = 0.05 is 3-84.
(c)  Significantly different from  the expected value at Station 1
     (P 1 0.05).

Note:  For all stations combined,  the calculated x2 with 4 Df = 10.93
       (PR > X2 = 0.028).
                                  D-13

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  TABLE D-7  LIST OF FISH  SPECIES AND FAMILIES COLLECTED SCIPPO  CREEK,
 	       AUGUST 1Q82(jO	
    Family
Cyprinidae
  (minnows)
Catostcmidae
  (sucker)
Centrarehidae
  (sunfish)
Percidae (perch)
    Scientific Name

Notropis photogenis

Notropis chrysocephalus

Semotilus atromaculatus

Rhinichthvs atratulus

Notropis spilopterus

Pimephales notatus

Ericvmba buccata

Campostoma anomalum

Notropis stramineus

Catostoraus commersoni

Hvpentelium nigricans

Ambloplites rupestris

Micropterus dolomieui

Etheostoma blennioides

E. caeruleum

E. flabellare

E. nigrum

E, zonale
     Common Name
Silver shiner

Striped shiner

Creek chub

Blacknose dace

Spotfin shiner

Bluntnose minnow

Silver jaw minnow

Stoneroller

Sand shiner

White sucker

Northern hog sucker

Rock bass

Smallmouth bass

Greenside darter

Rainbow darter

Fantail darter

Johnny darter

Banded darter
(a) Names follow  Robins  et  al.  1980.
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                                    D-14
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