United States
Environmental Protection
Agency
Environmental Research
Laboratory
Duluth MN 55804

Permits Division
(EN-336)
Washington DC 20460
EPA/600/3-86/006
July 1986


OWEP 86-03
Research and Development
Validity of
Effluent and Ambient
Toxicity Tests for
Predicting Biological
Impact, Kanawha
River, Charleston,
West Virginia

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                                   EPA/600/3-86/006
                                         July 1986
    Validity of Effluent and
  Ambient Toxicity Tests for
Predicting  Biological  Impact,
 Kanawha River, Charleston,
          West Virginia
                 Edited by
                        f
               Donald I. Mount
                  and
              Teresa Norberg-King

          Environmental Research Laboratory
         U.S. Environmental Protection Agency
              6201 Congdon Blvd.
            Duluth, Minnesota 55804
        Environmental Research Laboratory
        Office of Research and Development
       U.S. Environmental Protection Agency
             Duluth, MN 55804

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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 endorsement or recommendation
for use.

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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 Toxicity Testing Program are
  1.  To investigate the validity of effluent toxicity tests to predict 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 serve as a practical case example of how such testing procedures can be
     applied to effluent discharges in receiving water; and

  4.  To field test short-term chronic toxicity tests involving the test organisms,
     Ceriodaphnia dubia 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 guidelines. 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 necessity, be based on site-
specific water quality considerations.

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.

The following study was on the Kanawha River near Charleston, West Virginia,
and was conducted in August and September, 1984.

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

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 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 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 EffluentToxicity
                                                Testing Program
                                  IV

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                              Contents
                                                                  Page
Foreword
List of Figures
List of Tables
Acknowledgments
List of Contributors
Executive Summary
Quality Assurance
1.   Introduction	 1-1
2.   Study Design	2-1
3.   Site Description	3-1
4.   Laboratory Toxicity Tests	4-1
5.   Zooplankton Community Survey	5-1
6.   Periphyton Community Survey	6-1
7.   Macroinvertebrate Community Survey	7-1
8.   Comparison of Laboratory Toxicity Test Data and Receiving
    Water Biological Impact	8-1
References.
R-1
Appendix A:  Toxicity Test and Analytical Methods	A-1
Appendix B:  Biological Sampling and Analytical Methods	B-1
Appendix C:  Toxicity Test Data	C-1
Appendix D:  Biological Data	D-1

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                          List of Figures
Number
Page
   3-1   Study area and station locations on the Kanawha River,
        August and September 1984	3-1

   5-1   Number of zooplankton and macroinvertebrate taxa at various
        stations, Kanawha River	5-5

   8-1   Percent toxicity (using September data) vs. percent reduction
        in taxa	8-4

   8-2  Number of young per female and number of zooplankton taxa
        at various river stations	8-5
                                  VI

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                           List of Tables
Number                           Title                          Page

  3-1    Effluent Discharges to the Kanawha River	3-2

  3-2   Kanawha River Station Locations and Descriptions	3-2

  4-1    Young Production and Adult Survival of Ceriodaphnia Exposed
        to Various Concentrations of Fifteen Effluents,
        Kanawha River, August 1984	4-2

  4-2   Mean Individual Weight of Larval Fathead Minnows After Seven
        Days Exposure to Various Concentrations of Four Effluents in
        Upstream Water, Kanawha River, August 1984	4-4

  4-3   Seven-Day Percent Survival of Larval Fathead Minnows to
        Various Concentrations of Four Effluents in Upstream Water,
        Kanawha River, August 1984	4.5

  4-4   Results of Ambient Toxicity Tests with Ceriodaphnia,
        Kanawha River, Charleston, West Virginia, August 1984	4-6

  4-5    Mean Individual Weights of  Larval Fathead Minnows After
        Seven Days from Ambient Toxicity Tests of the Kanawha River,
        August 1984	4.7

  4-6    Seven-Day Percent Survival of Larval Fathead Minnows
        Exposed to Various Ambient Stations of the Kanawha River,
        Charleston, West Virginia, August 1984	4-8

  4-7    Ambient Toxicity Test Results with Ceriodaphnia, Kanawha
        River, Charleston, West Virginia, September 1984	4-8

  4-8    Mean Individual Weights (mg) of Larval Fathead Minnows
        After Seven Days from Ambient Toxicity Tests, Kanawha
        River, Charleston, West Virginia, September 1984	4-9

  4-9    Seven-Day Percent Survival  of Larval Fathead Minnows
        Exposed to Various Ambient Stations of the Kanawha River,
        Charleston, West Virginia, September 1984	4-9

  4-10 Acceptable Effluent Concentration (AEC) for 15 Effluents to the
        Kanawha  River	4.9

  4-11  Kanawha  River Flows, Charleston, West Virginia. Source of
        Data is the U.S. Geological Survey, Charleston,
       West Virginia	4-10

  5-1   Density of Zooplankton Collected from Various Ambient
       Stations of the Kanawha River, Charleston, West Virginia,
       September 1984	5-1
  6-1    Replicate Chlorophyll a, Biomass, and Autotrophic Index
       Values for Periphyton Collected from Artificial Substrates in
       the Kanawha River, Charleston, West Virginia,
       September 1984	6-1
                                   vii

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                       List of Tables fcont'd)

Number                           Title                           Page

  6-2    Mean Chlorophyll a and Biomass Standing Crops and
        Autotrophic Index Values for Periphyton Collected from
        Artificial Substrates in the Kanawha River, West Virginia,
        September 1984	6-2

  7-2    Numbers of Macroinverjtebrates from Artificial Substrates in
        the Kanawha River, Charleston, West Virginia in
        August 1984 	7-2

  8-1    Number of Taxa, Number of Ceriodaphnia Young per Female
        and Fathead Minnow Weights with the Associated Percent
        Reduction Using the Highest Value of Each as Zero Percent at
        Various Stream Stations (RK), Kanawha River	 8-4

  8-2    Percent of Stations Where Reduction in the Number of Taxa
        was Correctly Predicted by Toxicity Tests Using Four Arbitrary
        Levels of Comparison	8-4

  C-1   Water Chemistry Data for Effluent Toxicity Tests. Values are for
        Both Ceriodaphnia and  Fathead Minnow Tests and Final
        Dissolved Oxygen Values for the Daphnids Only	C-1

  C-2    Final Dissolved Oxygen [Concentrations for Fathead Minnow
        Larval Growth Tests onlEffluents, Charleston, West Virginia,
        August 1984	i	C-3

  C-3    Initial Water Chemistry Data for Ambient Toxicity Tests with
        Ceriodaphnia and Fathead Minnows on Day 1 of Testing,
        Charleston,  West Virginia, August 1984.	 C-3

  C-4    Final Dissolved Oxygen Concentrations for Ambient Toxicity
        Tests with Ceriodaphnia and Fathead Minnow,
        West Virgina, August 1984	 C-4

  C-5    Water Chemistry Data for Ceriodaphnia and Fathead Minnow
        Toxicity Tests, Kanawha River, Charleston, West Virginia,
        September 1984	 C-5

  D-1    Routine Chemistry Data for August and September for the
        Stream Stations. Readings Were Taken When Artificial
        Substrates Were Set and Removed, Kanawha River	D-1
                                 VIII

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                         Acknowledgments
The assistance and helpfulness of the State of West Virginia's Department of
Natural Resources was greatly appreciated. They collected samples along the
river, and assisted in setting the substrates. We  would especially like to
recognize Eli McCoy and Janice Fisher for their efforts during all stages of the
study. The assistance of the industries in collecting the effluent and obtaining
access to company grounds is acknowledged. EPA Region III, Wheeling, West
Virginia assisted in the study site selection  and pre-study coordination. The
efforts of Floyd Boettcher, Environmental Research Laboratory-Duluth field
engineer, in on-site responsibilities and sample collection is acknowledged.

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                         List of Contributors
                      LABORATORY TOXICITY TESTS
                Donald I. Mount8, Teresa J. Norberg-Kinga,
                    Scott Heinritz3, and Jeffrey Denny3
                         PLANKTON COMMUNITY
                 Sharon K. Gross", Randall B. Lewis0, and
                             Alexis E. Steenb
                        PERIPHYTIC COMMUNITY
       Randall B. Lewis0, Ronaldjj. Bockelmand, and Sharon K. Gross13
              BENTHIC MACROlNVERTEBRATE COMMUNITY
                   Randall B. Lewis", Thomas H. Rousha,
                  Alexis E. Steen0, and Sharon  K. Gross0
           COMPARISON OF LABORATORY TOXICITY DATA AND
                 RECEIVING WATER BIOLOGICAL IMPACT
                             Donald I. Mount3
               PRINCIPAL INVESTIGATOR: Donald I. Mount3
'U.S. Environmental Protection Agency, Envirorimental Research Laboratory-Duluth, 6201 Congdon Blvd., Duluth,
 MN 55804.                       !
6EA Engineering, Science and Technology, Inc., Hunt Valley/Loveton Center, 15 Loveton Circle, Sparks, MD
 21152.                          ;
CEA Science and Technology, 612 Anthony Trail, Northbrook, IL 60062.
"EA Engineering, Science and Technology, Inc., 221 Oakcreek Drive, Westgate Park, Lincoln, NE 68528.

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                        Executive Summary


 EPA recently issued a policy which provides for control of the discharge of toxic
 substances through the use of numerical criteria and effluent toxicity limits in
 IMPDES permits. This 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 study was the eighth in a series of eight and was conducted on the Kanawha
 River near Charleston, West Virginia, which receives discharges from  many
 industrial and municipal facilities. The study area comprises about 125 km of the
 Kanawha River, from the London Pool downstream to the Winfield Pool. The
 Kanawha River is an inland waterway and is navigable throughout the study
 area. Ambient toxicity  tests using both the Ceriodaphnia dubia and fathead
 minnow 7-d tests were conducted on samples from 34 river stations. Because of
 the nature of the  site, a comparison of ambient toxicity to community impact
 only, was attempted. Effluent dilution toxicity tests using Ceriodaphnia were
 conducted on samples from 11  discharges and fathead minnow effluent dilution
 toxicity tests were run on four  discharges. These effluent tests were  not a
 planned part of the study to meet the objective but were done to provide data to
 the West Virginia DNR. Biological studies conducted at the ambient stations
 included plankton, periphyton, and benthic macroinvertebrates.

 From 60 to 100% correct  predictions of community impact were made by the
 toxicity tests, depending on the levels of effect compared. There was a high (P <
0.005) correlation between Ceriodaphnia toxicity measured and  impact  on
 zooplankton over 125 kilometers of river, evidence that  the ambient test is an
 accurate  predictor of water quality effects on the instream biota. Impacts on
 macroinvertebrates  was underestimated by the ambient  tests. The toxicity
values derived from the effluent dilution tests do not suggest that the effluents
should cause toxicity after mixing.
                                  XI

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                         Quality Assurance


Coordination of the study was do,ne by the principal investigator preceding the
field work. A reconnaissance trip was made to the site prior to onsite work to
obtain the necessary details regarding each discharge and to make a cursory
evaluation of the river. Following that trip, details were delineated for setting the
sampling and testing dates and the specific sampling sites, as well as the specific
measurement to be made for each stream station. Upon arrival a meeting was
held with West Virginia's  Department  of Natural Resources, the principal
investigator, and the contract laboratory people to make final arrangements for
sampling of effluents and river sites. Also, the selection of effluents to be tested
was done. Following the meeting, a boat trip to identify the sampling stations and
to select where the artificial substrates would be placed was made. The principal
investigator was responsible for all the quality assurance related decisions. All
instrumentation used  during the study were calibrated  daily according to
manufacturers specifications.
                                   XII

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                                       1.     Introduction
 The study site was the Kanawha River near Charles-
 ton, West Virginia. The study area receives  many
 discharges from various industrial facilities. The study
 site was chosen to provide an opportunity to deter-
 mine if  ambient and effluent toxicity test results
 would correspond to the response of the biota in a
 large river. Toxicity was evaluated using  two test
 species. The response of the community was meas-
 ured using artificial substrates, as well as periphyton
 and zooplankton collection. The effluent dilution and
 ambient  toxicity tests were run on site where the
 substrates were placed in the river, and another set of
 ambient  toxicity tests  were  run  with river water
 collected when the substrates were removed.

 Several of the stations were  located in the zone of
 effluent mixing. The discharges and dilution volumes
 were so large that dye studies were too expensive for
 the funds available. The Kanawha River is channel-
 ized for ship and barge traffic and, without elaborate
 dye studies, the effluent concentrations at various
 stations  cannot be  approximated.  Therefore, the
 effluent dilution test results cannot be used to predict
 where impact should occur because the instream
 waste concentrations of each or any effluent are not
 known. However, using effluent flow discharge data
 and river flow, instream waste concentrations can be
 calculated. The river flow variation was large when
 the substrates were in place, and again there was no
 information as to how the flow affected the effluent
 concentrations at the sample stations where mixing
 was not complete. Thus, the effluent exposure those
 substrates experienced before and after the toxicity
 test period may have been the same as,  or  quite
 different  from, the exposure concentrations during
 the two periods  that ambient toxicity test  samples
 were collected.

 Determining the impact of individual discharges to
 rivers as large as the Kanawha is very difficult unless
 the impact  is dramatic. However, the combined
 effects of many discharges could be quite large, even
though any single discharge might have unmeasur-
 able effects on the aquatic community. Thus, the
value of  any  method that can estimate such indi-
vidually immeasurable impacts is obvious.

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

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                                      2.     Study Design
 Study components included (1) 7-day Ceriodaphnia
 dubia toxicity tests using samples from 34 ambient
 river stations, (2) 7-day larval fathead minnow growth
 tests  using samples from 34 ambient river stations,
 (3) eff Iuent tests of both species on selected eff Iuents,
 and (4) assessment of the zooplankton and benthic
 macroinvertebrate communities. Two separate sets
 of toxicity tests were conducted.The first set of tests
 was done on site and included both effluent dilution
 and grab ambient toxicity tests. The  second set of
 tests was done off site on shipped grab samples only
 on the same ambient stations as were tested while on
 site. These tests were run in the mobile trailer at the
 Environmental Research Laboratory-Duluth, Minne-
 sota.  The on site  ambient tests were done using 7
 different daily grab samples while the off site tests
 used a single grab sample for the entire test. In some
 instances insufficient sample was available for the
 latter series.
2.1   Toxicity Testing Study Design
Toxicity tests were performed on the effluents to
measure subchronic effects on the growth of larval
fathead minnows and chronic reproductive effects on
Ceriodaphnia (Chapter 4). A range of effluent concen-
trations was used so that acute mortality could be
measured as well as chronic mortality. The objective
of these tests was to estimate the minimum concen-
tration of each effluent  that  would cause  acute
mortality or chronic effects.

In addition to the effluent tests, ambient river stations
were selected and samples collected from them were
used to measure ambient toxicity to Ceriodaphnia and
fathead minnows (Chapter 4). These tests measured
the loss of toxicity from the effluents after mixing,
dilution from other  inputs, degradation, an'd other
losses such as sorbtion. These test results would also
provide data for the prediction of ecological impact for
comparison with the biological survey data, without
having to know the effluent concentration.

The off site  ambient toxicity tests were conducted
using samples collected during a period of low river
flows. These tests were done to see if the fungus
problem in the first set of tests had subsided and to
examine changes in toxicity due to lower river flow.
2.2   Biological Survey Study Designs
The field surveys included a quantitative assessment
of the zooplankton, periphyton, and artificial  sub-
strate macroinvertebrate communities. Artificial sub-
strates  were used to collect both periphytic and
macroinvertebrate organisms. The zooplankton data
are summarized in Chapter 5. Chlorophyll and bio-
mass were measured on periphyton (Chapter 6) and
the number of taxa and abundance  were measured
on the macroinvertebrates (Chapter 7).


2.3  Approach To Integration  of Labor-
atory and Field Efforts
The final component of this study was to integrate the
ambient toxicity  predictions with  the  measured
community impact. The results of the ambient toxicity
tests  can be used  to  predict community impact
regardless of whether instream waste concentrations
are known. The effluent tests were  done to provide
data to the West Virginia DNR and the data were not
used to predict effluent effects.
                                              2-1

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                                     3.     Site Description
The  Kanawha  River flows  northwesterly from its
origin at the confluence of the Gauley River and New
River in West Virginia  to the Ohio River. The study
area covers 125 km of the river length encompassing
rural and urban areas. All but two sampling stations
were located in the  London, Marmet and Winfield
Pools (Figure 3-1). Stations 20.1 and 25.7 were in a
pool formed by a dam on the Ohio River. River flow is
controlled at each of these three locks and dams.
                                          Eleven discharges were included in the study from
                                          river kilometer (RK) 67.1 toRK 143.5. The discharges
                                          were from diverse chemical and industrial facilities
                                          (Table 3-1). Ambient river stations for community
                                          surveys and toxicity testing were located from RK
                                          20.1 to RK 145.0. They were selected based on their
                                          relationship to effluent discharges in the river and
                                          were situated in near shore  areas generally away
                                          from barge traffic. Table 3-2 contains a listing of river
Figure 3-1.    Study area and station locations on the Kanawha River, August and September 1984.
                              ,20.1
                                                      105.2
                                                       108.9
                                                        D,C,B
                                                  112.0
                                                   114.2
                                                        119.1 125.5
                    Winfield
                    Lock & Dam
                                                        Blaine Island
                                                        Elk River
                        Marmet Lock & Dam
> Dischargers
• Ambient Stations
O POTWs(3)
                                                LONDON LOCK 8 C
Gauley
River

New
River
                                                3-1

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Table 3-1 . Effluent Discharges to the Kanawha River
River Secondary Industrial
Effluent Kilometer Category Codes
A 143.5
B 111.0
C 111.0
D 1 1 0.2
E 88.2

F 85.8
G 85.8

H 85.8

1 77.4
J 68.9

K 67.1


3313

2869, 2879
2869, 2879
2819

2812,2819


2812,2891
2819,2869,2879


2819,2869,2879


2869, 2879
2819,2869


2861,2869,2879,7391




Category Description
Electrometallurgical products
Industrial organics, agricultural chemicals
Industrial organics, agricultural chemicals
Industrial inorganic chemicals
Alkalies and chlorine, industrial inorganic
chemicals
Alkalies and chlorine adhesives and solvents
Alkalies and chlorine, industrial organics.
agricultural chemicals
Alkalies and chlorine, industrial organics.
agricultural chemicals
Industrial organics, agricultural chemicals
Industrial inorganic chemicals, industrial
organics
Gum and wood chemicals, industrial organics,
agricultural chemicals, research and
development laboratory
kilometers, dischargers and sampling stations. Only
those dischargers whose effluents were tested;are
listed but there were others. In addition, some of the
ambient  stations were   pairs  located  on  opposite
banks of the river.                                f
The navigational channels in the Kanawha  River are
maintained by dredging, so that shallow watersjare
only found very near shore. The locks and dams aljpng
the river influencethe habitat as does flow regulation.
During the on site testing in August, 1984, the river
was  quite high  due  to  intense rain  in the  up|per
watershed. Ambient testing was delayed several days
to allow the flow to  return to a  more normal onej
Table 3-2.

     River
   Kilometer
Kanawha River Station Locations and
Descriptions

             Station Description
      20.1     Biological sampling station; ambient station |
      25.7    Biological smapling station; ambient station I
      51.8    Biological sampling station; ambient station
      61.3    Biological sampling station; ambient station I
      66.OL   Biological sampling station; ambient station '
      66.0R   Biological sampling station; ambient station
      66.8    POTW 3 discharge
      67.1     K discharger
      68.4    Dilution water obtained for effluents J, K, arid
                POTW 3; biological sampling station; ambient
                station
      68.9    J discharger
      71.1     Biological sampling station; ambient station;
      76.1 L   Biological sampling station; ambient station'
      76.1 R   Biological sampling station; ambient station
      77.4    I discharger
      80.5    Dilution water obtained for effluent I; biological
                sampling station; ambient station        !
      83.5    Ambient station
      84.5    Biological sampling station; ambient station |
                                              Table 3-2.    (Continued)
                                                   River
                                                 Kilometer
                      Station Description
 85.8    H, G, F discharger; Davis Creek
 86.1     Dilution water obtained for effluents G and F
 87.4L   Biological sampling station; ambient station
 87.4R   Biological sampling station; ambient station
 88.0L   Biological sampling station; ambient station
 88.0R   Biological sampling station; ambient station
 88.2    E discharger
 89.6    POTW 2 discharge
 90.4    POTW 11 and POTW 1M discharges; Dilution
           water obtained for effluents E, H, POTW 2,
           POTW 1 M, and POTW 11; biological sampling
           station; ambient station
 92.5L   Biological sampling station
 92.5R   Biological sampling station; ambient station
 94.1     Biological sampling station; ambient station
 99.1     Biological sampling station; ambient station
101.4    Biological sampling'station; ambient station
105.2    Biological sampling station, ambient station
108.9    Biological sampling station; ambient station
110.2    D discharger
111.0    C, B discharger
112.0    Biological sampling station; ambient station
114.2    Dilution water obtained for effluents D, C, and B;
           biological sampling station; ambient station
118.4    Biological sampling station
119.1     Ambient station
125.5    Biological sampling station; ambient station
133.2    Biological sampling station; ambient station
139.0L   Biological sampling station; ambient station
139.0R   Biological sampling station; ambient station
142.7    Biological sampling station; ambient station
143.5    A discharger
145.0    Dilution water obtained for effluent A; biological
         sampling station; ambient station
                            3-2

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                               4.     Laboratory Toxicity Tests
 The purpose of the toxicity tests was to determine the
 ambient toxicity of water samples collected near the
 artificial substrates and to compare that toxicity with
 the response of the substrate community at each
 station. The ambient stations were chosen based on
 their location to an industry or group of industries to
 determine the effects on the river. Since flows were
 large and the number of discharges were too numer-
 ous to allow  dye  studies for  estimating instream
 waste concentrations, there was no critical need to do
 effluent dilution tests. Therefore, the selection of
 effluents for testing was left to the West Virginia
 Division of  Water Resources and  the choice was
 based on the needs of West Virginia's staff.
 Because the river was at a very high water stage
 during the August testing period, a set of ambient
 samples was collected in September and shipped to
 ERL-D to obtain measures of ambient toxicity at lower
 flows  (Table 4-11). During the onsite  testing,  a
 problem with a fungal growth on the Ceriodaphnia
 affected the effluent dilution tests but not the ambient
 tests. This fungus was clearly not parasitic because
 the animals  molted regularly and after molting, their
 appearance  was normal  until time passed and the
 fungus grew again. The effect was apparently only a
 physical one; the fungus weighed down the animals
 until they could  not remain  in the water column. In
 another  project  being done simultaneously in the
 same  mobile laboratory,  and using aliquots of the
 same  samples and test animals from  the  same
 culture, no fungus was observed. The only obvious
 difference was a 24-48 hour storage of the sample
 before use. Also, one industry split the effluent and
 dilution water samples collected for the on site  tests
 and had the  identical Ceriodaphnia 7-d tests done by
 a contract laboratory. That laboratory had no problem
 with fungus, and again, the sample had aged a few
 hours  during transit before  the tests were  started.
 Even the one percent effluent test solution greatly
 reduced  or eliminated the fungus growth.  For ex-
 ample, the  mean  survival  for the Ceriodaphnia
 dilution water controls  for the  15 effluent  dilution
 tests  was 41.5  percent (21.9  S.D.), whereas the
 survival for  the 1  percent  treatments for the 15
 effluents was  83.1  percent (16.6 S.D.). The mean
 young  per female and standard deviation was  16.1
 (3.7) and 21.3 (2.9) for the dilution water control and 1
 percent treatment,  respectively.  The fungus  was
 definitely not caused by the effluents tested in this
study.
 A similar problem was encountered in the Scippo
 Creek Study (Mount and Norberg, 1985); and again in
 water that was  shipped to the  offsite  lab  (and,
 therefore, was 24 hours older), there was no fungal
 growth.

 On two occasions previous to the site study, ambient
 samples were shipped from the Kanawha River to a
 remote  laboratory  and  no fungus problem was
 encountered.  Possibly, the fungus was  associated
 with the runoff following the rainfall which occurred
 just before the study began. Because of the rains, the
 ambient tests were started two days after the effluent
 dilution tests  and the fungus problem was minor
 substantiating that fungus was associated with the
 high flow.

 Brood size did not seem to be much affected by the
 fungus. If  the Ceriodaphnia did not get so "over-
 weighted" that they  died from struggling to free
 themselves, they produced normal broods. When test
 solutions   were changed,  those adults that were
 severely ladened were killed rather than  allowing
 them to die and the death being attributed to toxicity.
 These were stop-gap measures in order to obtain
 something from the tests. In the  September study
 using shipped river water samples, reproduction and
 survival  was excellent and  no sign of the fungus
 problem could be seen. Whether that was due to the
 delay caused by shipping or lower flow of the river is
 not known.

 The fathead minnows (Pimepha/esprome/as) did very
 well in both studies. During the onsite testing, the
 final  dissolved oxygen (DO) in the fathead minnow
 chambers  was about one  third  to one  half the
 concentrations of final DO in the  September tests.
 Both the ambient and the  effluent dilution tests
 showed this low DO in August and there appears to
 be no major  difference between  upstream  and
 downstream stations. Apparently  there  was an
 increased oxygen demand in the river associated with
 the high flows.
4.1   Chemical/Physical Conditions
4.1.1  Onsite Tests
Table C-1 contains the initial chemistry data for
effluent dilution tests for both test species, although
effluent dilution tests with the fathead minnows were
done only on four effluents. The final DO values are
                                               4-1

-------
for the Ceriodaphnia tests only while the final DO
values for the fathead minnow effluent tests are in
Table C-2. Table C-3 contains initial chemistry data
for the ambient tests and both species. Since the
effluents  were diluted  with water from various
ambient stations throughout the study area, initial
values were not taken on all stations after the first
day. The river velocity and turbulence was high, the
stations used for dilution water were close to one
another and a  decision was made  to  reduce [the
workload since differences between  stations un|der
such conditions were unlikely. Table C-4 contains jthe
final DO values for ambient tests on both species.
                                           i
All values for the Ceriodaphnia tests are in acceptable
ranges. The final DO values for the fathead minnows
are low and below the normally accepted range. Poor
growth was expected but, as will be shown lajter,
growth was excellent. The most probable reason is
thatthe DO was measured with a probe 1 cm or more
below the surface, whereas the fatheads were living
in the oxygen-rich surface film where DO was mUch
higher. Temperature for the  Ceriodaphnia test >lvas
25±1°C  and for the fathead minnow tests it was
25±3°C.
                                           I
4.7.2  Offsite Tests
Table C-5 contains the initial pH, DO, and conductivity
of the samples used for the ambient tests done offsite.
Since the entire test was done on the same sample
andthe samples were refrigerated between changes,
 initial chemistry was done only once.  Final DO values
for both species, done daily, are also in Table C-5J For
only  two stations and only for the fathead minnow
tests, are the DO ranges below 5.0  mg/L and then
 only  4.6 mg/L at the minimum. Nearly all values are
 in the acceptable range. Temperatures were 25±1 °C
for the Ceriodaphnia and 25±2°  C  for the fatrjead
 minnow tests.                              !
 4.2  Toxfcity Test Results
 Table 4-1 contains the results of the effluent dilution
 tests using Ceriodaphnia and Tables 4-2 and; 4-3
 contain the data for the fathead  minnow  effluent
dilution tests. In Table 4-1, the last column ("Number
of Test Animals") is the number of original animals
upon which the percent survival is based. Some of the
animals that were heavily fungused were intention-
ally removed to avoid their deaths being attributed to
toxicity. Almost without exception, young production
per female was near or at the normal 20 young per
adult expected (Mount and Norberg, 1984) for the
lowest two or three effluent concentrations.

Fathead minnow survival and growth was normal and
there was no fungal problem encountered. The 3
percent concentration of discharge A had poor growth
and survival for unknown reasons (Table 4-3). Only in
the effluents POTW1I  and POTW2, 100  percent
concentrations, were there  statistically lower growth
rates of the fathead minnows.

Table 4-10 contains the point estimates of acceptable
effluent concentrations (AEC) for both species and all
effluents tested. These AEC values are the geometric
mean of the no observed effect concentration (NOEC)
which causes no  adverse effect and  the  lowest
observed effect concentration (LOEC) which causes
an  adverse effect. Only three  effluents had effect
concentrations below 10  percent while  all  other
effluent AEC's were higher. None of the effluent
concentrations reached the AEC  after complete
mixing at the river flows existing during the study
because river flows were above  normal and the
effluent  flows were not  nearly large  enough to
produce instream waste concentrations (IWC's) ap-
proaching the AEC's.

Table 4-4 contains the on  site ambient test data for
the Ceriodaphnia and Tables 4-5 and 4-6 contain the
fathead minnow data. Stations at RK 145.0, 92.5R,
and 88.0L had statistically lower young production
than the station with the highest young production
value, RK 112.0. In addition, a small effluent-ladened
tributary, Davis  Creek, at  RK 85.8 had statistically
was lower young production. For the fathead min-
nows, survival was not lower (P<0.05) at any station,
but growth was reduced at RK 99.1, 87.4R, 87.41,
and 76.1 R as did Davis Creek (RK 85.8) as compared
to the station which hadthe the highest weight value,
 Tablo 4-1.   Young Production and Adult Survival of \Ceriodaphnia Exposed to Various Concentrations of Fifteen Effluents,
            Kanawha River, August 1984
Effluent
(RK)"'
POTW1I
(90.4)
Mean Number
Percent ofYoun^j Confidence
Effluent (v/v) per Female Interval
100 2.2lbl
30 20.9
10 20.0
3 22.5
1 24.7
Dilution Water
(90.4) 21.8
0.0-4.8
1 9.2-22.5
9.0-31.0
20.4-24.6
22.4-27.0
17.7-25.9
Percent
Survival
50
75
100
100
90
60
Number
of Test
Animals
10
8
4
4
10
10
                        4-2

-------
Table 4-1 . (continued)
Effluent Percent
(RK)"" Effluent (v/v)
POTW 1M
(90.4)





POTW 2
(89.6)





POTW 3
(66.8)





A (143.5)






6(111.0)






C (11 1.0)





0(110.2)






E (88.2)






100
30
10
3
1
Dilution Water
(90.4)
100
30
10
3
1
Dilution Water
(90.4)
100
30
10
3
1
Dilution Water
(68.4)
100
30
10
3
1
Dilution Water
(145.0)
100
30
10
3
1
Dilution Water
(114.2)
100
30
3
1
Dilution Water
(114.2)- 	 —
100
30
10
3
1
Dilution Water
(114.2)
100
30
10
3
1
Dilution Water
(90.4)
Mean Number
of Young
per Female
	 tb>
	 Ibl
22.0
19.5
21.1

17.3
	 (b)
	 (b>
24.2""
21.4
21.5

13.9
	 (b)
14.1
14.1
17.6
19.0

16.2
28.1lb)
21.0
24.8
18.8
20.2

18.3
Ib)
18.6
18.9
19.2
21.1

20.9
(b)
5.3
20.6
18.9

	 14.8
18.5
16.9""
19.3""
1 6.0""
1 6.6(b)

8.5
	 (b)
	 
-------
Table 4-1.    (continued)
Effluent
(RK)""
F (85.8)






G (85.8)






H (85.8)






1 (77.4)






J (68.9)






K (67.1 )






Percent
Effluent (v/v)
100
30
10
3
1
Dilution Water
(90.4)
100
30
10
3
1
Dilution Water
(86.1)
100
30
10
3
1
Dilution Water
(86.1)
100
30
10
3
1 .
Dilution Water
(80.5)
100
30
10
3
1
Dilution Water
(68.4)
100
30
10
3
1
Dilution Water
(68.4)
Mean Number
of Young
per Female
	 Ib)
Ib)
18.4
20.3
20.9

18.3
	 
-------
Table 4-2. (continued)
Effluent (RK)"" Replicate
POTW 1Mlbl
(90.4)

POTW 2""
(89.6)

A(bl
(143.5)

A
B
C
D
Weighted Mean
SE
A
B
C
D
Weighted Mean
SE
A
B
C
D
Weighted Mean
SE
Percent Effluent (v/v)
100
0.430
0.481
0.409
0.504
0.459
0.036
0.422
0.367
0.306
0.367
0.369|C)
0.035
0.649
0.793
0.540
0.598
0.645
0.041
30
0.563
0.583
0.457
0.458
0.516
0.035
0.478
0.511
0.426
0.515
0.481
0.033
0.467
0.711
0.511
0.692
0.587
0.041
10
0.489
0.520
0.526
0.584
0.527
0.036
0.383
0.501
0.517
0.559
0.486
0.032
0.615
0.736
0.556
0.650
0.639
0.041
3
0.521
0.643
0.535
0.517
0.548
0.037
0.542
0.651
0.539
0.608
0.582
0.033
0.357
0.390
0.424
0.495
0.413"='
0.050
1
0.563
0.544
0.597
0.678
0.596
0.036
0.469
0.690
0.550
0.618
0.578
0.034
0.584
0.644
0.701
0.677
0.648
0.043
Dilution
Water
0.589
0.734
0.431
0.538
0.571
0.036
0.724
0.565
0.546
0.602
0.609
0.035
0.553
0.628
0.558
0.640
0.596
0.041
'"'River kilometer of the discharge.
""POTWs, 11, 1M, and 2 were diluted with RK 90.4 water; A was diluted in RK 145.0 water.
(olSignificantly lower from each test's dilution water weights (P<0.05).
Table 4-3.    Seven-Day Percent Survival of Larval Fathead Minnows to Various Concentrations of Four Effluents in Upstream
              Water, Kanawha River, August 1984
Percent Effluent (v/v)
Effluent (RK)'al
POTW 11""
(90.4)



POTW1M"3'
(90.4)



POTW 2""
(89.6)



Albl
(143.5)



Replicate
A
B
C
D
Mean
A
B
C
D
Mean
A
B
C
D
Mean
A
B
C
D
Mean
100
70
90
50
80
72.5
90
100
80
100
92.5
90
100
70
70
82.5
80
100
100
100
95
30
90
100
90
100
95
90
100
100
90
95
90
80
100
100
92.5
100
90
100
80
92.5
10
100
100
100
100
100
100
90
100
80
92.5
100
100
100
80
95
100
90
90
100
95
3
100
100
100
100
100
90
70
80
100
85
100
100
100
60
90
70
70
50
60
62.5
1
80
80
80
80
82.5
90
90
100
90
92.5
80
70
100
90
85
90
100
80
70
85
Dilution
Water
80
80
100
100
90
100
90
100
80
92.5
80
70
90
80
80
90
100
90
90
92.5
'"'River kilometer of the discharge.
""POTWs, 11, 1 M, and 2 were diluted with RK 90.4 water; A was diluted in RK 145.0 water.
Note: No significantly lower differences for any eflfuents were found (P < 0.05).
                                                                                       4-5

-------
Tabta 4-4.   Results of Ambient Toxioity Tests with Cerio-
           daphnia, Kanawha River, Charleston, West
           Virginia, August 1984
Ambient Mean Number
Station Young Per
(RK) Female
145.0
142.7
139.0R
139.0L
133.2
125.5
119.1
114.2
112.0
108.9
105.2
101.4
99.1
94.1
92.5R
90.4
88.0R
88.0L
87.4R
87.4L
Davis Creek, 85.8
83.3
84.5
80.5
76.1 R
76.1 L
71.1
68.4
66.0R
66 OL
61,3
51,8
25,7
20,1
15.9"11
18.0
19.6
21.3
19.0
17.1
17.9
18.7
24.8
16.7
17.9
16.0
18.5
21.1
15.0""
19.0
22.7
	 (a)
19.9
20.6
6.13'"
24.3
16.5
19.6
19.2
19.8
19.5
21.6
23.4
17.2
16.4
17.4
19.3
20.5
Confidence Mean Percent
Intervals Survival
13.1-18.6
14.9-21.1
14.5-24.6
16.8-25.6
12.3-25.6
13.6-20.6
14.9-20.9
16.0-21.4
22.3-27.3
12.4-21.1
1 2.4-23.3
10.7-21.6
14.3-22.8
16.1-26.0
12.5-17.5
17.0-21.0
19.7-25.7
—
1 5.3-24.5
1 8.2-23.0
2.7-9.4
22.5-26.1
12.1-20.9
1 5.2-24.0
14.6-23.8
1 6.2-23.4
14.4-24.6
17.0-26.2
20.3-26.7
11.9-22.5
14.3-18.4
12.8-22.0
16.2-22.4
17.6-23.4
80
100
70
80
78
88
89
100
90
70
70
90
80
80
90
90
100
0
80
100
60
80
100
100
100
90
100
90
90
100
89
89
100
100 \
 "Significantly different (P < 0.05).
 RK 88.0L. However,  none of those stations were
 greatly different from the highest value they were
 compared to even though they  were statistically
 different. Such differences could be due to ot)ier
 causes such as enriched water with more food. For
 the Ceriodaphnia at RK 88.0L and Davis  Creek (RK
 85.8), differences in feeding level  would  not be
 expected to cause such low numbers. The food used
 will consistently produce.20 young  in reconstituted
 water (Mount,  unpublished data), where the food
 added is the only food available. The differences in
 Ceriodaphnia production at Stations 145.0, 92.5R,
 and 61.3 could be the result  of experimental varia-
 tions or food level. Tables 4-7, 4-8, and 4-9 contain
 the ambient test data for the September  testing on
 single grab samples shipped to Duluth. There were no
 statistically significant differences  in survival and
 growth or reproduction for either species when the
 station with the highest value  is used for comparison.
 Survival of Ceriodaphnia was notably  low  at RK
stations 125.5,108.9, and 84.5, although not statis-
tically significant. The method of analyses for young
per female (Appendix A) essentially excludes effect of
adult mortality  on young production estimates. In
general, the reproduction of Ceriodaphnia and growth
of the fathead minnows was uniform and at slightly
above levels normally obtained in unenriched water.

4.3  Discussion
Since the low concentration of effluents, as well as
the higher ones, eliminated  the fungus problem, the
data can be  used  but with some caution.  Quality
control sets  should have been included in which a
known water of good quality was used. Since samples
had  been tested previous  to this study and  good
performance was obtained, they were not thought to
be needed. Si nee the effect of the fungus would be to
overestimate toxicity, and the AEC values obtained
are all higher than the instream waste concentrations
(IWCs) (for the fathead minnows as well as Cerio-
daphnia), one can conclude that the effluents should
not cause toxicity  at the flows existing during the
study.
The  on site  ambient tests with Ceriodaphnia, in
general, had acceptable survival and, whereas there
were five stations with significantly reduced young
production, only Station 88.0L and Davis Creek (RK
85.8) are below the normal range usually obtained.
The values that are significantly lower for the fathead
minnows are also within the  normal range. Therefore,
the toxicity, if any, is certainly not very great. Several
spills were  reported during the study period by
various plants in the study  area  and, since the test
animals were exposed to a new sample every day,
some effect of these spills could be evidenced by
these data.
No statistical differences were  found in the Sep-
tember study for either species. The overall impres-
sion from both  testing periods is that  the effluents
tested are not causing toxicity after dilution at the
flows prevailing during the  two study periods.  If the
reduced growth of the fathead minnow and young
production of the Ceriodaphnia is due to toxicity, it is
minimal.

The oxygen demand of the ambient water during the
August period  of  high  flow, and the widespread
fungus problem associated with it, perhaps should be
further investigated. Fungal growths  were also found
on the artificial substrates providing some field
evidence as well that the problem observed in the
tests was not just an artifact.
                        4-6

-------
Table 4-5.    Mean Individual Weights of Larval Fathead Minnows After Seven Days From Ambient Toxicity Tests of the Kanawha
              River, August 1984.                                                        ;
Ambient Station
(RK)
145.0
142.7
139.0R
139.0L
133.2
125.5
119.1
114.2
1 1 2.0
108.9
105.2
101.4
99.1
94.1
92.5R
90.4
88.0R
88.0L
87.4R
87.4L
Davis Creek, 85.8
84.5
83.3
80.5
76.1 R
76.1 L
71.1
68.4
66.0R
66.0L
61.3
51.8
25.7
20.1

A
0.527
0.503
0.526
0.431
. 0.547
,0.462
0.542
0.503
0.415
0.462
, 0.548
0.551
0.492
0.574
0.491
0,507
0.586
0.610
0.382
0.520
0.388
0.486
0.580
0.498
0.610
0.490
0.552
0.599
0.682
0.452
0.520
0.568
0.572
0.532
us
' B
. 0.595
0.593
0.566
0.536
0.671
0.512
0.524
0.575
0.533
0.537
.' 0.579
0.584
0.456
0.674
0.578
0.467
0.505
0.659
0.490
0.465
0.443
0.550
0.543
0.572
. 0.441
0.548
0.561
0.424
0.492
0.490
: 0.445
0.467
0.536
0.546
spncaie
C
0.432
0.336
0.419 .
0.521 *
0.462
0.532
0.386
0.502
0.583
0.528
0.420
0.565
0.500
0.454
0.575
0.430
0.493
. 0.536
0.432
0.271
0.457
0.563
0.498
0.361
0.374
0.517
0.576
0.497
0.456
0.562
0.439
0.496
O.572
i 0.607

D
0.528
0.490
0.485
• 0.441
0.536
0.601
0.775
0.623"
0.450
0.463
0.622
0.451
0.322
0.517 .
0.458
0.588
0.522
0.623
: 0.460
0.484 .
0.393
0.546
0.442 '
0.479
0.393
0.479
0.594
0.438:
0.644
0.490
0.574
0.504
0.493
0.689
Weighted
Mean
0.521
0.492
0.489
. 0.485 •
0.554
0.523
0.504*
' 0.546
0.501
0.494
0.553
0.542
0.443""
0.554
0.523
0.498
0.526
0.603
: 0.441 lal
0.435""
0.420""
' 0.536
0.516
0.478
. 0.455lal
0.509
0.571'
0.490
0.569
0.499
0.495
0.509
0.543
0.594
SE
0.038
0.038
0.036
0.036
0.038
0.037
0.041
0.038
0.036
0.035
0.036
0.034
0.036
0.033
0.034
0.033
0.034
0.036
0.033
0.033
0.033
0.033
0.033
0.033
0.033
0.033 •
0.033
0.034
0.034
0.034
0.034
0.035
0.035
0.034
""Station 88.0L was used for comparisons, significantly lower (P < 0.05)
                                                                                     4-7

-------
Table 4-6.    Seven-Day Percent Survival of Larval Fathead
              Minnows Exposed to Various Ambient Stations
              of the Kanawha River, Charleston, West Virginia,
              August 1984
Table 4-7.   AmbientToxicityTestResultsWithCe/yo
-------
 Table 4-8.
               Mean Individual Weights (mg) of Larval Fathead Minnows After Seven Days from AmbientToxicity Tests, Kanawha
               River, Charleston, West Virginia, September 1984
Ambient Station
(RK)
145.0
142.7
139.0R
139.0L
133.2
125.5
119.1
112.0
108.9
101.4
94.1
92.5L
90.4
88.0L
87.4L
84.5
76.1
68.4
66.0R
61.3
51.8
25.7
20.1
Note: No significantly

A
0.606
0.575
0.588
0.669
0.590
0.622
0.606
0.569
0.635
0.585
0.633
0.505
0.578
0.645
0.550
0.617
0.572
0.645
0.500
0.450
0.611
0.620
0.617
Hepl
B
0.615
0.630
0.533
0.567
0.615
0.622
0.630
0.550
0.539
0.544
0.678
0.570
0.550
0.689
0.675
0.600
0.494
0.681
0.629
0.689
0.600
0.717
0.595
Itcate
C
0580
0.675
0.550
0.450
0.539
0.564
0.656
0.567
0.560
0.644
0.645
0.750
0.469
0.750
0.656
0.522
0.520
0.528
0.605
0.550
0.605
0.517
0.670
lower differences for any stations were found (P < .05).

D
0.517
0.544
0.625
0.550
0.610
0.564
0.705
0.525
0.515
0.694
0.644
0.695
0.531
0.461
0.714
0.520
0.680
0.556
0.511
0.658
0.570
0.611
0.557
Station 94.1

Mean
0.581
0.608
0.571
0.560
0.590
0.597
0.649
0.552
0.563
0.616
0.649
0.620
0.534
0.637
0.648
0.565
0.567
0.605
0.539
0.577
0.596
0.619
0.614
was used for comparison.

SE
0.023
0.023
0.024
0.025
0.023
0.025
0.023
0.023
0.023
0.033
0.023
0.038
0.037
0.038
0.038
0.037
0.036
0.037
0.044
0.034
0.033
0.033
0.033

 Table 4-9.    Seven-Day Percent Survival of Larval Fathead
              Minnows Exposed to Various Ambient Stations
              of the Kanawha River, Charleston, West Virginia,
              September 1984
Ambient Station
(RK)
145.0
142.7
139.0R
1 39.0L
133.2
125.5
119.1
112.0
108.9
101.4
94.1
92.5L
90.4
88.0L
87.4L
84.5
76.1
68.4
66.0R
61.3
51.8
25.7
20.1
A
100
100
89
80
100
90
90
80
100
100
100
100
100
100
90
90
100
100
90
100
90
89
90
Replicate
B
100
100
90
90
10
90
100
100
90
90
90
100
100
90
100
100
90
100
88
90
80
100
91
C
100
100
100
80
90
70
90
100
100
90
100
80
100
90
90
90
100
100
90
73
100
90
100
D
90
90
80
70
100
70
90
100
100
90
90
80
80
90
78
100
100
80
90
60
100
90
70
Mean
Survival
98
98
90
80
98
80
93
95
98
93
95
90
95
93
90
95
98
95
90
81
93
92
88
Table 4-10.   Acceptable Effluent Concentration (AEC)"" of
              15 Effluents for the Kanawha River
                                                                                     AEC (Percent Effluent [v/v])
Effluent
POTW1
POTW 1
POTW2
POTW 3
A
B
C
D
E
F
G
H
I
J
K
Ceriodaphnia
54.8
17.3
17.3
54.8
>100
54.8
17.3
>100
5.5
17.3
54.8
5.5
17.3
17.3
5.5
Fathead Minnow
54.8
>100
54.8

>100










                                                            '"'Geometric mean of the no observed effect concentration (NOEC)
                                                             which causes no adverse effect and the lowest observed effect
                                                             concentration (LOEC) which causes an adverse effect.
Note: No significantly lower differences for any stations were
     found (P < .05). Station 94.1 was used for comparison.
                                                                                     4-9

-------
Table 4-11.
Kanawha River Flows, Charleston, West
Virginia. Source of Data is the U.S. Geological
Survey, Charleston, West Virginia
                   Date
                                       Flows (m3)
August 13
August 14
August 15
August 16
August 17
August 18
August 19
August 20
August 21
August 22
August 23
August 24
August 25
August 26
August 27
August 28
August 29
August 30
August 31
September 1
September 2
September 3
September 4
September 5
September 6
September 7
September 8
September 9
September 10
September 1 1
September 12
September 13
September 14
September 1 5
September 16
September 17
Mean flow during onsite testing
Mean flow during substrate exposure
Mean flowduringzooplankton sampling
750.5
1 243.3
1 081 .8
589.1
382.3
259.4
317.2
253.2 ;
185.5 :
184.4
244.4
230.5
175.9
140.5
113.3
91.5
104.5
130.3
235.6
869.4
55.1
266.8
303.0
262.2
253.8
188.6
172.2
182.1
124.3
99.7
92.3
109.3
102.8
103.7
127.2
122.0
301 .6
202.1
115.5
                             4-10

-------
                         5.0     Zooplankton Community Survey
The zooplankton community was sampled using the
methods described in Appendix B. The current of the
Kanawha River was fast (even though the river is
totally in pools as a result of navigation dams) during
the August onsite study as a result of rain upstream.
Flows were from 1243 to 253  m3/sec during the
sampling period (August 14-20),  about four to 20
times above  normal low  flow (see Table 4-11).
Samples for zooplankton were collected in August
when the substrates were placed in the  river, but
upon examination  no  further counts were  made
because there were insufficient densities to be valid.
Samples were taken again from 12-17 September
when the artificial substrates were removed and after
the flow had been at more normal summer values.


5.1   Zooplankton  Populations
Table 5-1 lists the taxa and density of organisms for
the three replicate samples at each station. Density at
RK 20.1, 25.7, 51.8 and 61.3 are up to three times
greater than some of the other upstream stations and
there is an abrupt drop in density at the stations just
upstream of RK 61.3. This change is  not reflected in
the number of taxa which is the same and at the
 maximum for RK 51.8, RK 61.3, RK 66R and RK 68.4.
 There is no conspicuous reason for a drop in density.
 RK  66 is over 40 kilometers downstream  of the
 Marmet Dam with no obvious change in the river in
 that  reach.  The  Pocatalico  River enters between
 stations 61.3 and 66.0.

 The trend in number of taxa  is shown in Figure 5-1
 which shows a slight downward trend from down-
 stream to upstream, excepting the two lower most
1 stations (these two had a higher density). Tributary
 inputs to the  mainstream Kanawha River are rel-
 atively small, the stream is totally in pool between the
 dams and a decreasing  stream size  or shorter
 residence time does not seem a likely cause. The 50
 percent or more decrease in number of taxa from
, Stations 51.8, 61.3, 66L and 66R and 68.4 to Station
 87.4L and 87.4R to 108.9 is certainly not a result of
 stream size or residence time.

 While the change in density from downstream  to
 upstream is bigger than in number of taxa, both show
 a similar trend.
Table 5-1.   Density of Zooplankton Collected from Various Ambient Stations of the Kanawha River, Charleston, West Virginia,
           September 1984 (Number per 100 mi)

Brachionus calycifloris
Brachionus quadradentatus
Euchlanis sp.
Platyias quadricornis
Lecane sp.
Asplanchna sp.
Diaphanosoma sp.
Daphnia sp.
Ceriodaphnia sp.
Bosmina sp.
llyocryptus sp.
Chydorinae
Camptocercus sp.
Alona sp.
Diaptomus sp.
Cyclops sp.
Encyclops sp.
Leptodora kindtii
Branchiura
Total taxa per station

A
0
0
0
0
0
0
10.5
2.0
0
29.5
0.5
0
0
1.5
76.0
12.5
0
0
0

20.1
B
0
0
0
0
0
0
8.5
2.0
0
25.5
0.5
0
0
0.0
31.5
15.5
0
0
0
7

C
0
0
0
0
0
0
1.0
1.0
0
12.5
1.0
0
0
1.5
18.5
9.0
0
0
0


A
0
0
0
0
0
0
8.5
.0.5
0
'3.5
1.0
0.5
0
2.0
7.5
7.0
0
0
0

25.7
B
0
0
0
0
0
0
5.0
1.0
0
8.5
0.5
0
0
0.5
27.5
8.5
0
0
0
7

C
0
0
0
0
0
0
3.5
0.5
0
5.5
0.5
0
0
1.0
16.0
9.0
0
0
0


A
0
0
0
0.5
0
0
21.0
0.5
0
3.5
0
0
0
0.5
6.0
6.5
2.5
0
0

51.8
B
0
0
0
0
0
0
20.5
0.5
0
2.5
0
0
0
0.5
5.0
5.5
1.5
0
0
10

C
0
0
0
0
0
0.5
15.0
1.0
0
4.0
0.5
0
0
0
3.0
2.5
1.0
0
0


A
0
0
0.5
0
0
1.5
4.5
0.5
0
8.5
8.0
0
0
4.5
4.0
24.5
12.0
0
0

61.3
B
0
0
0
0
0
0
1.5
0
0
2.0
1.0
0
0
1.0
2.0
1.5
4.5
0
0
10

C
0
0
0
0 '
0
o
5.0
0
0
1.0
0
0
0
0.5
3.0
0.5
2.5
0
0

                                              5-1

-------
Table 5-1. (Continued)


Brachionus calyciflor/s
Brachionus quadradentatus
Euchlanls sp.
Platyias quadricornis
Lecane sp.
Asptanchna sp.
Diophanosoma sp.
Daphnia sp.
Ceriodaphnia sp.
Bostnina sp.
Hyocryptus sp.
Chydor/nae
Camptocercus sp.
Alona sp.
Dfaptomus sp.
Cyclops sp.
Eucyclops sp.
Leplodoro kindtii
Branchiura
Total taxa per station
Table 5-1. (Continued)


Brachionus calyciflor/s
Brachionus quadradentatus
Euchlanis sp.
Platyias quadricornis
Locana sp.
Asptanchna sp.
Diaphanosoma sp.
Daphnia sp.
Ceriodaphnia sp.
Bosmina sp.
Hyocryptus sp.
Chydorfnae
Camptocercus sp.
Alona
Diaptomus sp.
Cyclops sp.
Eucyclops sp.
Leptodora kindtii
Branchiura
Total taxa per station


A
0
0
1.0
0
0
0
1.0
0
0
0.5
0
0
0
1.0
1.5
2.5
2.5
0
0



A
0
0
0.5
0
0
0
0.5
0
0
1.0
0
0
0
1.5
1.5
0
2.5
0
0


66.0L
B
0
0
0.5
0
0
0
2.0
0
0
1.5
1.0
0
0
0
1.0
2.5
1.0
0
0.5
9

76.1 L
B
0
0
0
0
0
0
0
0
0
0
0
0
0
0
2.0
0
1.5
0
0
7


C
0
0
0.5
0
0
0
0.5
0
0
•1.5
: 0
0
0
3.0
0.5
1.5
1.5
0
0


i
IC
0
0
0
0
0
0
0.5
0
0
0
0.5
0
0
0
1.0
0
4.0
0
0



A
0
0
0
0
0
0
1.0
0
0
2.0
0
0
0
0
2.5
0
1.5
0
0.5



A
0
0
0.5
0
0
0
0
0
0
0.5
0.5
0
0
0.5
1.0
0
2.5
0
0.5


66.0R
B
0
0
2.0
0
0
0
5.0
0.5
0
1.5
0
0
0
0
2.0
1.5
2.5
0
0
10

76.1 R1
B
0
0
0
0
0
0
0
0
0
0.5
0
0
0
0
2.5
0
0.5
0
0
7


C
0
0
0.5
0
0
0.5
1.0
0
0
0
0
0
0
0.5
2.5
0
1.5
0
0.5



C






















A
0
0.5
1.0
0
0
0
0.5
0
0
'2.0
0
0
0
0
1.5
1.0
2.0
0
0



A
0
0
0
0
0
0
0
2.5
0
0
0
0
0
0
2.0
0
1.5
0
0


68.4
B
0
0
0.5
0
0
0
1.0
0
0
0.5
0
0
0
0
1.5
1.5
1.5
0
0.5
10

80.5
B
0
0
0.5
0
0
0
0
2.0
0
0
0
0
0
0
0.5
0.5
0.5
0
0.5
7


C
0
0
0.5
0
0
0
1.0
0.5
0
3.5
0
0
0
0.5
3.0
1.5
1.5
0
0



C
0
0
0
0
0
0
0.5
3.5
0
0
0
0
0
0
0
0
0
0
0



A
0
0
0
0
0
0
0.5
0
0
1.0
0
0
0
2.0
1.5
0.5
3.5
0
0



A
0
0
0
0
0
0
0.5
0
0
0
0
0
0
0.5
0
1.0
3.0
0
0


71.1
B
0
0
0
0
0
0
0
0
0
0
0
0
0
0.5
1.0
0
2.5
0
0
7

84.5
B
0
0
0
0
0
0
0.5
0
0
0
0.5
0
0
0
0.5
1.5
0.5
0
0
7


C
0
0
0
0
0
0
0
0
0
0
0.5
0
0
0.5
0.5
0
2.0
0
0



C
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0.5
0.5
0.5
0
0

'Replicates A and C were totaled together and listed under A, due to error in counting.
                             5-2

-------
Table 5-1. (Continued)


Brachionus calycif Ion's
Branch/onus quadradentatus
Euchlanis sp.
Platyias quadricornis
Lecane sp.
Asplanchna sp.
Diaphanosoma sp.
Daphnia sp.
Ceriodaphnia sp.
Bosmina sp.
llyocryptus sp.
Chydorinae
Camptocercus sp.
Alona sp.
Diaptomus sp.
Cyclops sp.
Eucyclops sp.
Leptodora kindtii
Branchiura
Total taxa per station
Table 5-1. (Continued)


Brachionus calycif loris
Brachionus quadradentatus
Euchlanis sp.
Platyias quadricornis
Lecane sp.
Asplanchna sp.
Diaphanosoma sp.
Daphnia sp.
Ceriodaphnia sp.
Bosmina sp.
llyocryptus sp.
Chydorinae
Camptocercus sp.
Alona sp.
Diaptomus sp.
Cyclops sp.
Eucyclops sp.
Leptodora kindtii
Branchiura
Total taxa per station


A
0
0
0
0
0
0
0
0
0
1.0
0.5
0
0
0
0
0
1.0
0
0



A
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0


87.4L
B
0
0
0
0
0
0
0.5
0
0
0
0.5
0
0
0.5
0
0
1.0
0
0
5

90.4
B
0
0
0.5
0
0
0
0
0
0
0
0
0
0
0
0
0.5
0
0
0
4


C
0
0
0
• o
0
0
0
0
0
0
0
0
0
0
0
0
1.5
0
0



C
0
0
0.5
0
0
0
O
0
0
0
0
0
0
0.5
0
0
2.0
0
0



; A
0.5
0
0.5
0
0
0
0
0
0
0
0
0
0
0
0
0
1.0
0
0



A
0
0
0.5
0
0
0
0.5
0
0
0
1.0
0
0
3.0
0
0
2.0
0
0


87.4R
B
0
0
1.0
0
0
0
0
0
0
0
0
0
0
0
0
0
0.5
0
0
3

92.5L
B
0
0
1.0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
5


C
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0



C
0
0
0
0
0
0
o
0
0
0
0
0
0
0
0
0
1.5
0
0



A
0
o
0
0
o
0
0.5
o
0
o
0
o
0
1.0
0
o
0
0
0



A
0
0
0
0
0
0
o
0
0
0
2.0
0
0
2.0
0
o
1.5
0
0


88.0L
B
0
0
0
o
0
0
0
o
0
o
0
o
1.0
0.5
0
o
0.5
o
0
5

92.5R
B
0
0
0.5
0
0
0
0
0
0
0
0
0
0
0.5
0
0
1.0
o
0
5


C
0
o
0
o
o
0
0.5
o
0
o
0
o
0
0.5
o
0.5
0
0
0



C
0
0
1.0
0
0
0
o
0
0
0
1.5
0
0.5
0
0
o
1.0
o
0



A
0
o
0.5
o
o
0
0
o
0
o
0
o
0
0
o
o
1.5
o
0



A
0
0
1.0
0
0
0
o
0
o
0
0
0
0
0.5
0
Q
1.5
o
0


88.0R
B
0
o
0.5
o
o
0
o

0
o
0.5
o
0
1.0
o
o
1.0
o
0
4

94.1
B
0
0
0.5
0
0
0
0
0
o
0
0
0
0
0.5
0
0.5
o
0
4


C
0
n
\j
1.0

0
o

0
1.0
o
2.5
o

2.5
0



C
0
0
1.0
0
0
0
0
o
0
0.5
0
0
0.5
0
1.5
0

5-3

-------
Table 5-1.   (Continued)


Brachionus calyciftoris
Brachionus quadradentatus
Euchlanis sp.
Platyias quadricornts
Locane sp.
Asptanchna sp.
Diaphanosoma sp.
Daphnia sp.
Ceriodaphnfa sp.
Bosmina sp.
llyocryptus sp.
Chydorinae
Camptocercus sp.
/4/ona sp.
Dioptomus sp.
Cyclops sp.
Eucyclops sp.
Leptodora kindtii
Branchiura
Total taxa per station
Table 5-1. (Continued)


Brachionus calycHloris
Brachionus quadradentatus
Euchlanis sp.
Platyias quadricornis
Locans sp,
Asptanchna sp.
Diaphanosoma sp.
Daphnia sp.
Ceriodaphnia sp.
Bosmina sp.
llyocryptus sp.
Chydorinae
Camptocercus sp.
/4/ono sp.
Dioptomus sp.
Cyclops sp.
Eucyclops sp.
Leptodora kindtii
Branchiura
Total taxa per station

A
-0
0
0
0
0
0
0
0
0
0
0.5
0
0
2.5
0
0
0.0
0
0.5



A
0
0
0.5
0
0
0
0.5
0
1.0
0
2.5
0
0
3.0
0
0
0
0
0

99.1
B
0
0
0
0
0
.0
0
0
0
0
1.5
0
0
0
0
0
0.5
0
0
4

112.0
B
0
0
0
0
0
0
0
0
0
. 0
1.0
0
0
1.5
0
0
0
0
0
7

C
0
,,0
0
0
0
0
0
0
0
', o
0
0
0
1.0
[ 0
0
1.5
0
0



C
0
! 0
(0.5
0'
0
0
0
1.0
0
0
1.0
0
0
1.5
0
0
1.5
0
0


A
0
0
0
0
0
0
0.5
0
'0
0
0
0
0
0
0
0
1.0
0
0



A
0
0
1.0
0
0
, 0
;o
0
0
0
0
0
0
1.0
0.5
0
1.0
0
0

101.4 >
B
0
0
0
0
0
0
o:
0
0
0
0
0
0
1.0
0.5
0
1.5
0
0
4

114.2
B •
0
0
0.5
0
0
• 0
0
0
0
0
0
0
0
0
0
0
0.5
0
0
5

C
0
0
0
0
0
0
0
'0
0
0
0
0
0
1.0
0.5
0
0
0
0



C
0
0
0
0
"o
0
0
0
0
0
0.5
0
0
0.5
0
0
0.5
0
0


A
0
0
0
0
0
0
0
0
0
1.0
0
0
0
0
0
0
0.5
0
0



A
0
0
1.5
0
0
0
0
0
0
0.5
0.5
0
0
1.5
0
0
0.5
0'
0

105.2
B
0
0
0.5
0
0
0
0
0
0
2.0
0.5
0
0
0.5
0
0
1.0
0
0
5

118.4
B
0
0
0
0
0
0
0
0
0
0
0.5
0
0
1.0
0
0
0
0
0
5

C
0
0
1.5
0
0
0
0
0
0
0.5
0
0
0
0.5
0
0
1.0
0
0



C
0
0
0
0
0
0
0
0
0
0.5
0.5
0
0
8.0
0
0
1.0
0
0


A
0
.0
0.5
0
0
0
5.5
0
0
0.5
0
0
0
2.5
0.5
0
1.5
0
1.5



A
0
0
0
0
0
0
0
0.5
0
0
0.5
0
0
0.5
0
0
0.5
0
0

108.9
B
: 0
, 0
0
0
0
0
1.5
0
0
1.5
0.5
0.,
0
1.5
0 .
0
1.0
0
1.5
8

125.5
B
0
0
0
0-
0-,
0 .,
0.5
1.5
0
" 0
0 •
6".
0
1.0
0
o'
0.5
0
0
7

C
0
0
0.5
0
0
0
2.5
0
0
1.0
0
0
0
1.5
0
0
1.5
0
0.5



C
0
o .-,
0.5
0
. 0
0
1.5
1.0
0
" 0
°'l>
, 0 '
0
0
0.5
0.5
-,0
0

                              5-4

-------
Table 5-1.    (Continued)

Brachionus calycif Ion's
Brachionus quadradentatus
Euchlanis sp.
Platyias quadricornis
Lecane sp.
Asplanchna sp,
Diaphanosoma sp.
Daphnia sp.
Ceriodaphnia sp.
Bosmina sp.
llyocryptus sp.
Chydorinae
Camptocercus sp.
Alona sp.
Diaptomus sp.
Cyclops sp.
Eucyclops sp.
Leptodora kindtii
Branchiura
Total taxa per station

A
0
0
0
0
0
0
5.0
0
0
0
0
0
0
0
0
0
0.5
0
0

133.2
B
0
0
0.5
0
0
0
1.5
0
0
0
0.5
0
0
0.5
0
0
0.0
0
0
6

C
0
0
1.0
0
0
0
2.0
0
0
0
0
0
0
0
0
0
0.5
1.0
0


A
0
0
0
0
0
0
0
0
0
0.5
0
0
0
0
0
0.5
0
0
0

1 39.0L
B
0
0
0
0
0
0
0
0
0
0
0
0
0
0
2.5
0
0.5
0
0
7

C
0
0
1.0
0
0.5
0
0
0
0
0
0.5
0
0
0
0
o
1.5
0
0


A
0
0
0.5
0
0
0
0
0
0
0
0.5
0
0
0
0.5
0
1.0
0
0

139.0R
B
o
o
0.5
0
0
0
0
o
0
o
0
o
0
o
0.5
o
0.5
0
0
4

C
o
o
0
o
0
o
o
o
0
o
0
o
0
o
0.5
o
0.5
o
0


A
o
o
0
o
1.0
o
o
o
o
o
0.5
o
o
1.0
0
o
0.5
o
0

142.7
B

Q
0
o
0.5
n
\j
o
n
\j
0
n
\j
0
Q
o

0.5

0
o
0
5

C

n
u
0
o
1.0
n
\j
Q

o

0.5
o

o

0.5
Q
0

Table 5-1.    (Continued)
145.0

Brachionus calycif loris
Brachionus quadradentatus
Euchlanis sp.
Platyias quadricornis
Lecane sp.
Asplanchna sp.
Diaphanosoma sp.
Daphnia sp.
Ceriodaphnia sp.
Bosmina sp.
llyocryptus sp.
Chydorinae
Camptocercus sp.
Alona sp.
Diaptomus sp.
Cyclops sp.
Ducyclops sp.
Leptodora kindtii
Branchiura
Total taxa per station
A
0
0
0.5
0
0.5
0
0
0
0
0
0
0
0
0
0
0
0
0


B
0
0
0
0
1.0
0
0
0
0
0
0
0
0
0
0
0
1.5
0

3
C
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1.5
0


                                                            Figure 5-1.    Number of zooplankton and macroinvertebrate
                                                                          taxa at various stations, Kanawha River.
                                                             30
                                                           •S20
                                                           J3

                                                           3
                                                             10
                                                                    20
                                                                                                       O     00
        o  o o     o
00        8
     o    o
                   .Zooplankton
                   0 Macroinvertebrates
                                                                           40
                                                                                 60     80    100
                                                                                   River Kilometer
                    120   140   160
                                                                                    5-5

-------

-------
                             6.     Periphyton Community Survey
 6.1   Chlorophyll a and Biomass
 Measurements

 Samples for chlorophyll a and biomass were collected
 on 12-16 September 1984. The artificial substrates
 had been submerged at varying water depths due to
 large changes in river stage caused by the early rains.
 Appendix B describes the sampling techniques.

 Variability was large both between  replicates at a
 station and between stations (Table 6-1). In general,
 higher values for chlorophyll a  were obtained at
 Station 99.1 and upstream except for Stations 20.1
 and 25.7. Mean concentrations were > 1.6 mg/m2
 chlorophyll a in this reach of the  Kanawha  River,
 whereas downstream  of  Station  99.1, the  mean
 concentration for 40 percent of the stations was <0.5
 mg/m2 chlorophyll a (Table 6-2).

, Periphyton biomass, measured as ash-free dry weight
. (AFDW), varied from 0.051 to 8.577 g/m2(Table 6-1).
 Similar  to  the  chlorophyll a data, lowest values
 occurred at Station SS.OLand highest values occurred
 at Station 20.1. Such similarities are expected since
 chlorophyll  a and  AFDW  measure algal biomass.
Table 6-1.    Replicate Chlorophyll a, Biomass, and Autotrophic Index Values for Periphyton Collected from Artificial Substrates in
            the Kanawha River, West Virginia, September 1984
Station
(RK)
20.1
25.7
51.8
61.3
66.0L
66.0 R
68.4
71.1
76.1 L
76.1 R
80.5
84.5
87.4 L
87.4 R
88.0 R
88.0 L
90.4
92.5 L
92.5 R
94.1
99.1
101.4
105.2
108.9
112.0
114.2
118.4
125.5
133.2
139.0L
139.0R
142.7
145.0
Chlorophyll a (mg/m2)
A
73.168
0.352*
0.402
2.465
0.692
4.641
0.818
0.957
0.085*
3.678
0.554
0.048*
0.044*
4.778
0.253*
0.013*
0.211*
0.106*
0.486
0.851
0.869
--
22.026
8.189
2.308
16.270
3.326
1.240
4.021
3.629
1.231
15.538
14.096
B
13.794
49.281
1.325
1.349
'
2.626
0.143*
15.919
0.137*
2.373
0.623
0.057*
0.051*
0.089*
0.175*
0.007*
0.226*
0.550
0.093*
1.216
1.519
--
--
8.873
3.325
8.967
2.299
3.112
5.943
8.669
2.499
11.262
4.172
C
8.208
5.825
0.084*
0.996
--
0.789
0.147*
0.352*
0.335*
2.640
0.363
0.068*
—
0.258*
—
0.022*
--
0.024*
0.103*
--
12.508
--
--
9.872
1.370
24.994
3.073
0.457*
2.874
7.807
4.421
7.320
7.674
Biomass (g/m
A
8.577
0.577
0.551
1.165
1 .709
1.910
1.020
2.096
1.107
1.720
1.143
0.311
0.362
0.458 i
0.661
0.102
0.261
0.100
0.273
0.425
0.921
--
2.187
1.430
0.508
1.971
1.060
0.876
1.193
0.905
0.645
3.404
2.563
B
4.107
5.755
0.991
0.908
"
1.876
1.434
2.734
1.881
1.233
0.873
0.689
0.519
0.314
0.261
0.051
0.198
0.360
0.351
0.606
1.326
--
—
1.297
0.995
1.600
0.803
0.825
1.263
2.113
0.779
2.081
0.976
2)
C
3.121
1.844
0.695
0.904
--
1.948
1.746
1.201
2.550
1.220
1.086
0.444
—
0.052
--
0.150
--
0.054
0.198
"
2.289
--
—
1.568
0.666
5.039
0.766
0.486
1.024
1.287
0.898
1.447
1.538
Autotrophic Index
A
117
1,639
1,371
473
2,470
412
1,247
2,190
13,024
468
2,063
6,479
8,227
96
2,613
7,846
1,237
943
562
499
1,060
—
99
175
220
121
319
706
297
249
524
219
182
B
298
117
748
673
-.
714
1 0,028
172
13,730
520
1.401
12,088
10,176
3,528
1,491
7,286
876
655
3,744
498
873
__
-.
146
299
178
349
265
213
244
312
185
234
C
380
317
8,274
908
	
2,469
11,878
3,412
7,612
462
2,992
6,529
—
202
__
6,818
	
2,250
1.922
__
183
__
	
159
486
202
249
1,063
356
165
203
284
200
Notes: Asterisk (*) indicates chlorophyll a value based on fluorometric analysis.
     Dash (--) indicates artificial substrate(s) missing.
                                                6-1

-------
Mean values of AFDW vary from 0.101 to 5.268
g/m2; however, no trends are observed.

Values of an autotrophic index (Al) were calculated
following that of Weber (1973), and were based on
the ratio of AFDW to chlorophyll a. The Al values wep
> 160 at all stations except Station 105.2. Such high
Al values indicate that the periphyton community |is
dominated by either non-algal (heterotrophic) taxa or
nonliving organic matter.


6.2   Evaluation of the Periphytic
Community
High mean chlorophyll a values were found at the two
most downstream stations(20.1 and 25.7) and above
Station 94.1 (Table 6-2). Very high Al values for the
stations located between Stations 25.7 and 94.1
indicate that the periphyton community was either
non-algal or nonliving. The material causing the high
Al values could be the same as the  fungus-like
material observed  in the toxicity tests.

Table 6-2.   Mean Chlorophyll a and Biomass  Standing
            Crops and Autotrophic Index Values for
            Periphyton Collected from Artificial Substrates
            in the Kanawha River, West Virginia, September
            1984
Station
(RK)
20.1
25.7
51.8
61.3
66.0 L
66.0 R
68.4
71.1
76.1 L
76.1 R
80,5
84.5
87.4 L
87.4 R
88.0 R
88.0 L
90.4
92.5 L
92.5 R
94.1
99.1
101.4
105.2
108,9
112.0
114.2
118.4
125.5
133.2
139.0L
139.0R
142.7
145.0
Chlorophyll a
(mg/m2)
31.723
18.486
0.604
1.603
0.692
2.685
0.369
5.743
0.186
2.897
0.513
0.058
0.048
1.708
0.214
0.014
0.218
0.227
0.227
1.034
4.965
--
22.026
8.978
2.334
1 6.744
2.899
1.603
4.279
6.702
2.717
1 1 .373
8.647
Biomass
(9/m2)
5.268
2.725
0.746
0.992
1.709
1.911
1.400
2.010
1.846
1.391
1.034
0.481
0.440
0.275
0.461
0.101
0.230
0.171
0.274
0.516
1.512
~
2.187
1.432
0.723
2.870
0.876
0.729
1.160
1.435
0.774
2.311
1.692
Autotrohic
Index
265
691
3,464
685
2,470
1,198
7,718 '
1,925
11,455
483
2,152 ;
8,365
9,226
1,309
2,052 ;
7,317
1,056
1,283
2,086
498
705
--
99
160
335
167
306
678
289
219
346
229
205
 Note:  Dash (--) indicates artificial substrate was missing.

                         6-2

-------
                       7.     Macroinvertebrate Community Survey
The macroinvertebrates were measured using arti-
ficial substrate samples suspended for approximately
four weeks. The water samples for zooplankton and
toxicity testing were taken in close proximity to the
substrate samplers.  The substrates were placed
between 14-20 August and were recovered from 12-
17 September. They were located out of barge traffic
lanes and typically 5-20 meters from shore. Where
discharges occurred  but were not obviously fully
mixed, stations  on each  side of the river were
established. The collection techniques are described
in Appendix B.3.


7.1   Macroinvertebrate Populations
The number oftaxa (Table 7-1 and Figure 5-1) show a
rather clear increase from downstream to upstream.
The station with the highest (99.1) and lowest number
of taxa (87.4L) are less than 12 kilometers apart,
however.  The station (RK 87.4L)with  the lowest
number of taxa, 14, is located downstream of Elaine's
Island, an area of a high concentration of discharges.
The total number of organisms collected at RK 87.4
was much lower compared to Station 99.1.

The use of artif ical substrates reduces habitat effects.
Therefore, decreasing numbers downstream would
seem to be water quality caused. This is reinforced by
the close proximity of stations with  high and low
numbers of taxa.  In contrast to the zooplankton, the
four  most downstream stations are not  markedly
higher than the stations just upstream of these four
stations.

The trends shown by the macroinvertebrates are
definitely different than the trend of the zooplankton.
                                              7-1

-------
Tabla 7-1 . Numbers of Macroinvertebrates From Artificial Substrates in the Kanawha River, Charleston. West Virginia, August
1984
20.1 25.7 51.8 61.3

Stenonema
Caenis
Tricorythodes
Isonychis
Baetis
Nouroclipsis
Hydropsyche
Cheumatopsyche
Polycentropsis
Hydroptila
Small cased caddis
Holeidae
Atherix
Hemerodromia
Chironomus
Pseudochironotnus
Tribelos
Dicrotendipes
Glyptotendipes
Polypedilum
Micropsactra
Rhootanytarsus
Tanytarsus
Cricotopus
Psoctrocladius
Corynoneura
Nanocladium
Ablabesmyia
Labrundinia
Tanypus
Neohermes
Acroneuria
Optioservus
Argia
Didymops
Ostracoda
Hyalella
Gammarus
Physa
Gyrinus
Sphorium
Ferrissima
Hydracarina
Hydra
Hirudinae
Oligochaete
Planaria
Nematoda
Cryptochironomus
Procladius
Motrionemus
A
5
4
17

54


2
13


1



3
158
90
42
6
22
21
18
20
4
18







2

5






127





B
20
1
5

170



3






165
22
12
8
13
2
54
4
33

1







2


7



18

1



C A
15 19
4 2
5 1

58 21



2






7 78
125 226
118
10 3
3
4
9 1
12 1
1
19 10





1
1
1

2






14 12
2




B C A B C
26 14 4 20 12
6341
3342

54 43 38 47 89








1 2

67 14 19 7 35
135 272 148 181 135
11 32 25
68 6 5 11
3 15 12 4
32 21
4 5 10 8
10 2 16 15 14
1 1
19 28 24 39 23






1
3— i
7
1
1 1


1
1


7 5 14 23 28
2 2 1
2 21
1


ABC
10 8 14

11 2 3

56 1 57 56










45 99 43
15 12 2
544
8 51 8
3 1
342
8 26
2
7 15 10
















3 14 5





Total taxa per station
                                   24
                                                           21
                                                                                   20
                                                                                                            15
                            7-2

-------
Table 7-1. (Continued)
66.0L

Stenonema
Caenis
Tricorythodes
Isonychia
Baetis
Neureclipsis
Hydropsyche
Cheumatopsyche
Polycentropsis
Hydroptila
Small cased caddis
Heleidae
Atherix
Hemerodromia '
Chironomus
Pseudochironomus
Tribelos
Dicrotendipes
Glyptotendipes
Polypedilum
Micropsectra
Rheotanytarsus
Tanytarsus
Cricotopus
Psectrocladius
Corynoneura
Nanocladius
Ablabesmyia
Labrundinia
Tanypus
Neohermes
Acroneuria
Optioservus
Argia
Didymops
Ostracoda
Hyalella
Gammarus
Physa
Gyrinus
Spherium
Ferrissima
Hydracarina
Hydra
Hirudinae
Oligochaete
Planaria
Nematoda
Cryptochironomus
Procladius
Metrionemus
A B
4
2
2

55

1








64
13
22
4
3
33
3

17
1
















19
1




66.0R 68.4
CABCABCA
3222371
323 476
8893167

44 35 84 162 66 59 63
1
1


1
1



2 26 17 21
83 79 80 24 107 48 127
21 3 11 28 19
18 24 7 5 2 6 23
30 40 16 8 6 5 4
11 15 6 3
38 50 20 13 10 20
65 18 5 6
26 23 20 7 1 2 18
342 1

11 10 11 17 27 15 34
1




1
1
1




2 1


1

125 61 41 30 23 28 32
1
1



71.1
B
5
5
5
1
74





1



2
93
6
10
4
2
87
10
42
g
1
28




1

2









96

1




C
3
1
5

89









1
23
4
4
g

5
17
2

35
















60
1




Total taxa per station
                                   16
                                                           19
                                                                                   20
                                                                                                           24
                                                                                       7-3

-------
Tablo 7-1. (Continued)

Stenonema
Caonis
Tricorythodes
Isonychia
Baotis
Neuroclipsis
Hydropsyche
Chaumatopsyche
Polycantropis
Hydroptila
Small cased caddis
Holaidao
Atherix
Hemerodromia
Chironomus
PsQudochironomus
Tribelos
Dicrotendipes
Glyptotondipes
Polypedilum
Micropsectra
Rhootanytarsus
Tanytarsus
Cricotopus
Psactroclndius
Corynoneura
Nanocladius
Ablabesmyia
Labrundinia
Tanypus
Neohermes
Acroneuria
Optioservus
Argia
Didymops
Ostracoda
Hyalolln
Gommnrus
Physa
Gyprinus
Sphorium
Ferrissima
Hydracarlna
Hydra
Hirudinae
Oligochaete
Planaria
Nomatoda
Cryptochironomus
Procladius
Metrionemus
A
6
5
2

50









34
81
4
5
15
2
8
33





1
2
1








25





76.0L
B
7
3
1

96









6
36
4
5
1
19
1
18
1
41








1




2


68
3
3




C
1
1

43








2
36
49
6
9
2
33
2
8
36







1
1







52
3

1



A
6
3 !
1

32









1
89
9 !
17
7
28
2
3
11
2
16





1
1









64
3




76.0R
B C
9 4
3
11 4

28 94



1

1



1
151 50
5 8
16 15
15 8
3 8
39 16
8
22 19
9 1
18 33






1









148 83
2 2




80.5
A B
7 6
3 3
6 5

46 34









4
16 25
12
13 28
40 4
34 22
7 87
3 16
32 15
8 7
14 19
1







•i
i






75 119





84.5L
C A B C
12 3 2 5
4 12 5 8
9 1 4 12

30 23 23 33'





911
£. \ 1
1 ,

4 3 16
33 26 9 30
2
28 33 36 , 9
46 32 42 23
13 14 21 14
8 11 21 25
4827
33 5 8 5
6 54
14 6 11 12
21
i




1 1
i i









151 20 57 34

1
1


Total taxa per station
                                 24
                                                         21
                                                                                 19
                                                                                                        21
                           7-4

-------
Table 7-1. (Continued)
87.4L
ABC
Stenonema 11
Caenis 1 1
Tricorythodes 1
Isonychia
Baetis
Neureclipsis 24 43
Hydropsyche
Cheumatopsyche
Polycentropis
Hydroptila
Small cased caddis
Heleidae
Atherix
Hemerodromia
Chironomus
Pseudochironomus
Tribelos 1 2 1
Dicrotendipes 50 12
Glyptotendipes 2 2
Polypedilum 5 6
Micropsectra 22 5
Rheotanytarsus
Tanytarsus 16 4
Cricotopus 12 1
Psectrocladius
Corynoneura
Nanocladius
Ablabesmyia 7 10
Labrundinia
Tanypus
Neoriermes
Acroneuria
Optioservus
Argia 1
Didymops
Ostracoda
Hyalella
Gammarus
Physa
Gyprinus
Spherium
Ferrissima
Hydracarina
Hydra
Hirudinae
Oligochaete 29 8
Planaria
Nematoda
Cryptochironomus
Procladius
Metrionemus

A
7
7
11


24

3









5
3
gg
100
6
75
5
8
7

4

















18

1



87.4R
B
24
6
21

1
25

2




1



6
21

46
16
33
15
21

12

6

















24

1




C
16
14
8


8
1
3
1




1



17
3
70
22
25
48
12

7
2
8

















"22






A
58
11
16


40


1







8
15
2
14
75
18
6
2
5
5

g

















2

1



88.0R
B C
38
3
13


20

1
1



1
1


1
4
1
28
47
10
28
8

6

2

















8





88.0L
A B
1
2 3



16 17











17 2
98 53
9
5
2 6

16 9
1
1 1


5 16





1
1 1
1








7 16






C





16











12
77

2
18

19




4
















18





Total taxa per station
14
                        24
                                                                                   21
                                                                                                           16
                                                                                        7-5

-------
Tablo 7-1.    (Continued)

Stenonema
Caonis
Tricorythodes
Isonychia
Baotis
Neureclipsis
Hydropsyohe
Cheumatopsyche
Polycentropis
Hydroptila
Small cased caddis
Heleidao
Atherix
Hemorodromia
Chironomus
Pseudochironomus
Tribelos
Dicrotendlpes
Glyptotendipes
Polyp edilum
Micropsectra
Rheotanytarsus
Tanytarsus
Cricotopus
Psoctrocladius
Corynonoura
Nanocladius
Ablabesmyia
Labrundinia
Tanypus
Neohermes
Acroneuria
Optioservus
Argia
Didymops
Ostracoda
Hylolla
Gammarus
Physa
Gyprinus
Sphorium
Forrissima
Hydracarina
Hydra
Hirudlnae
Oligochaete
Planaria
Nematoda
Cryptochironomus
Procladius
Metrionemus

A
55
10
21


33







2




25

21
37
67
16
3
23
6

18
1



1

3









20
1




90.4
B C
36
6
12


14








1



24

34
41
8
48
21
2
17
5
3






1









17

2




A
8
12
17


16










1
10
42

12
47
27
35
1
3
12

18






1 I
1








53





92.5L
B
45
15
23


23


1








1
45

30
77
16
61
11
14
19

7
















90






C
62
6
16


24

1









1
19

68
43
14
73
9
14
12
3
1
1















12

2




A
14
44
22

2
13










4
37

11
18
21
122
13
14
6

9






1









1
247

1



92.5R
B C
10 9
22 12
12 7


25 36





1




30 45
31 53

10 12
12 53
13
26 9
1 5
4 4
8

18 21




4


1









16 34
4
1




A
7
9
6


20










9
49
5
9
27
12
69
4
11
3

7





2

1









114
4




94.1
B C
14
9
8


23










6
34
1
15
43
12
28
3
5
1

9






1
2









138
3




  Total taxa per station
22
                        23
                                                                                    23
                                                                                                            20
                             7-6

-------
Table 7-1. (Continued)

Stenonema
Caenis
Tricorythodes
Isonychia
Baetis
Neureclipsis
Hydropsyche
Cheumatopsyche
Polycentropis
Hydroptila
Small cased caddis
Heleidae
Atherix
Hemerodromia
Chironomus
Pseudochironomus
Tribelos
Dicrotendipes
Glyptotendipes
Polypedilum
Micropsectra
Rheotanytarsus
Tanytarsus
Cricotopus
Psectrocladius
Coryoneura
Nanocladius
Ablabesmyia
Labrundinia
Tanypus
Neohermes
Acroneuria
Optioservus
Argia
Didymops
Ostracoda
Hyalella
Gammarus
Physa
Gyprinus
Spherium
Ferrissima
Hydracarina
Hydra
Hirudinae
Oligochaete
Planaria
Nematoda
Cryptochironomus
Procladius
Metrionemus
A
26
8
3


29

1
1



2


2
11
51

23
113
7
37
32
12
6
1
4






1
1









29
3
1



99.1
B
11
2
3


27










20
80
1
1
50
1
21
8
4
3

18







3 .





2

2

89
2




101.4
CAB
31
4
11
1
3
30

2

5



3

3
3
60

60
90
23
31
24
11
9

5




1

1










58





105.2
CAB
52

17
1
3
16
25
13

5


2
1



14

93
79
33
21
37

14

3






1










4

2




C A
2
5
3

1
11











20
3
44
78
1
67
22
8
2

6
1






3



1



5

42
2




108.9
B
3
3
2


4



1






6
39
5
47
40
13
105
36
14
3

6
1






4



12

1



248
11





C

10
2

1
8










6
17
4
24
42

38
7
6


2







10
6

1
6





184
45




Total taxa per station
32
                                                                            21
                                                                                                             26
                                                                                         7-7

-------
Tablo 7-1. (Continued)
112

Stononoma
Casnis
Tricorythodes
Isonychta
Baotls
Neureclipsis
Hydropsyche
Choumatopsyche
Polycontropis
Hydroptila
Small cased caddis
Heleidae
Atherix
Hemorodromia
Chironomus
Pseudochironomus
Tribelos
Dicrotendipes
Glyptotendipes
Polypodilum
Micropsectra
Rheotanytarsus
Tanylarsus
Cricotopus
Psoctrocladius
Corynoneura
Nanocladius
Ablabesmyia
Labrundinia
Tanypus
Neohermes
Acroneuria
Optioservus
Argia
Didymops
Ostracoda
Hyalella
Gammarus
Physa
Gyrinus
Sphorium
Fornssima
Hydracarina
Hydra
Hirudinao
Oligochaete
Planaria
Nematoda
Cryptochironomus
Procladius
Metrionemus
A B
5 5
2 5
2


40 65










1

13
75 44
11
7 2
21 11
7
12 3
1
2
1

7 5



1
1


3 2




1
1



19 8



C A
12 7
5 4
2

2
35 3



4








16 5
87 94
1 14
16 32
2
10 43
8

5

8 11







9









19 6
2


114.2
B
7
3
6

2
10




2


1



1
13
123
2
21
61
34


2

12
1






13









11
1



C
6
3
1

1
4



1



1




4
91
7
17
17
30
4
2


6
1






24









10




A
28
8
8

1
•18

1










15
11
71
12
21

4


6
1






2









6



119.1
B
23
15
14


15












4
59
21
35
26
109
. 3
3

1
19



1



3









46




C
28
15
7


17

1
1

2







5
24
1
25
97
7
43
9
5
3
3
7







1




2



2
14

1


A
19
4
10

1
19










6
9
1
23
30
50
39
4
3
3

8

















13





125.5
B
19
3
10


15








1

7
61
1
40
123
10
34
13
3
3
5
8
2






2









22






C
24
6
12
•i
I

15










5
41
41
18
31
85
7
1
9

3







1



1
i




26





Total taxa per station
                                   22
                                                           25
                                                                                   27
                                                                                                           28
                           7-8

-------
Table 7-1. (Continued)

Stenonema
Caenis
Tricorythodes
Isonychia
Baetis
Neureclipsis
Hydropsyche
Cheumatopsyche
Polycentropis
Hydroptila
Small cased caddis
Heleidae
Atherix
Hemerodromia
Chironomus
Pseudochironomus
Tribelos
Dicrotendipes
Glyptotendipes
Polypedilum
Micropsectra ,
Rheotanytarsus
Tanytarsus
Cricotopus
Psectrocladius
Corynoneura
Nanocladius
Ablabesmyia
Labrundinia
Tanypus
Neohermes
Acroneuria
Optioservus
Argia
Didymops
Ostracoda ;
Hyalella
Gammarus
Physa
Gyrinus
Spherium
Ferrissima
Hydracarina
Hydra
Hirudinae
Oligochaete
Planaria
Nematoda
Cryptochironomus
Procladius
Metrionemus
A
6

3

3
2










25
32

13
43

20
5

2

12
2
1





42

1
6
8


1
1

129
51

2


133.2
B
3
2













1
14
41
3
21
17
7
28
2

2

6







9


1
6

4

7

129
7
1




C
4
1


1











13
23

15
30
14
15
13

1
1
4







9



6

8



172
4





A
4
1
6
1
1
7
1









2
61

26
66

34
9

2

16





1

3
1








17


2


139L
B
16
7
20
1
5
4










4
154

4
72

17


3

15







12
2








36






C
28
1
6


5


1





3

15
87

30
68

19
4
5
2
2
23





1

19





1



33
2
1
6


139R
A B
18 13

13 18
1
1
1 4


1 1





1 1

52 42
79 28
1
32 3
50 19
6
34 3
8
2
1

11 8




1
1

11 5




1
1



76
2
1
3



C
18

15


2


3







54
24

7
15

16
1
1


5







3









1
2





A
10

19
1
6
12









2
17
56

26
73
15
17
9
1
10

20
1

1

1


7





1



87





142.7
B
24

28
1
5
16







1


9
51

47
76
8
5
16
4
14

16







5.









73






C
38

33

4
7



1


1



3
80

29
75
8
35
9
4
13
6
15
1






7

1







35


3



A
16

10
1
9
1



2




1

36
97

30
19

20
15
3
2

14
1






6
5




1



36





145.0
B
25

9

1
4









1
13
108

6
18

22
3

1

18








1


22
1
2



12






C
23
3
9

4
2



1






38
153

17
41

18
3
2
1
1
26
1



1


12
4




1



50


4


Total taxa per station
30
                                                        27
                                                                     27
                                                                                        28
                                                                                                            28
                                                                                       7-9

-------

-------
8.     Comparison of Laboratory Toxicity Test Data and Receiving Water Biological
       Impact
The comparison between toxicity measured  in the
laboratory on a few species and the impact occurring
in the stream on whole communities must compen-
sate for a very limited database from which to predict.
The sensitivity of the test species relative to that of
species in the community is almost never known and
certainly not in these effluent toxicity tests. Therefore,
when toxicity is found, there is no method to predict
whether  many species in the community, or just a
few, will  be adversely affected at similar concentra-
tions, since the sensitivities of the species  in the
community are not known. For example, at a given
waste concentration, if the test species has a toxic
response and if the test species is very sensitive, then
only those species  in the community  of equal or
greater sensitivity  would be  adversely affected.
Conversely, if the test species is tolerant ofthe waste,
then many more species in the community would be
affected at  the concentration which begins to cause
toxic effects to the test species. It is possible that no
species in the community is as sensitive as the most
sensitive test species, but since there are so many
species composing the community, this is unlikely. It
is more  likely  that a  number of species in the
community will  be more  sensitive than the  test
species.  The highest probability is  that the  test
species will be near the median sensitivity of organ-
isms in the community if the test species is chosen
without knowledge of its sensitivity (as was the case
in this study).

In a special case, where toxicants remain the same
and the species composing the community remain
the same, the number of species in the community
having a sensitivity equal to or greater than the test
species also will remain the same. As a result, there
should be  a consistent relationship  between the
degree of toxicity as measured by the toxicity test and
the  reduction in the number  of species  in  the
community. In this special case,  there should be  a
tight correlation between degree of toxicity and the
number of species. If the toxic stress is great enough
to diminish the  production of offspring by a  test
species, it should also be severe enough to diminish
the reproduction of some species within the com-
munity of equal  or greater sensitivity. This should
ultimately lead to elimination of the more sensitive
species. Therefore, a lower number of taxa should be
a predictable response of the  community.  For ex-
ample, there should be a relationship between the
number of young per female Ceriodaphnia or the
growth of fathead minnows (or other test species) and
the number of species in the community. Obviously,
the test species must have a sensitivity, such that at
ambient concentrations to which the community has
responded, a partial effect is produced in the toxicity
test. However, unless the special case described
above exists, the correlation between toxicity  and
species richness will not be a tight one.

Effluents  differ from  single chemicals in  some
important  respects. We know from the literature on
single chemicals that there usually are large differ-
ences in  the  relative  sensitivity of  species  to  a
chemical and  that the relative sensitivity changes
with different  chemicals. For example, the fathead
minnow may be more sensitive to effluent A  and
Ceriodaphnia more sensitive to effluent B. We also
know that effluents vary in their composition from
time to time and often within a few hours. We should
not be surprised, therefore, to find fatheads being
more sensitive to an effluent on one day and daphnids
more sensitive on another day.

Effluents begin changing in composition as soon as
they are discharged. Fate processes such as bacterial
decomposition, oxidation and  many others change
the composition. In addition, various components will
change at  different rates.  For example, ammonia
would be  expected to disappear  more rapidly than
PCBs. If so, then the composition of the effluent is
ever changing as it  moves through the receiving
water. Note that this change is not just a lessening
concentration as a result of dilution but also a change
in the relative  concentrations of the components. In
reality, the aquatic organisms at some distance from
the outfall  are exposed to  a different  toxicant than
those near the discharge point! Therefore, it is logical
to expect that sometimes one test species would be
more sensitive to the effluent as it is discharged and
another species more sensitive after fate processes
begin altering the effluent. To be sure the source of
the effluent is the same but it is certainly not the same
"effluent"  in  regard to its composition. If these
statements are true then one should also expect that
species in  the community in the receiving water will
be affected at one place near the discharge and  a
different group of species will be affected from the
same effluent at another location.
                                               8-1

-------
Compound the above described considerations with
multiple discharges as well as inputs from tributaries
and non-point sources such as agricultural run-off
and leachate from landfills and one should logically
expect an unpredictable effect on various components
of the community.  Figure  5-1 shows two  clearly
different trends  between zooplankton and macrb-
invertebrates,  evidence of the above effects.

An effluent cannot be viewed as just diluting as it
moves away from the outfall. In fact, it is a "series of
new eff I uents" with elapsed flow time. If so, there are
important implications for interpretation of toxicity
and community  data. One should not expect the
various test species to respond similarly to water
collected from various ambient stations. We should
expect one species to be more sensitive at one station
and another species to be more sensitive at the next.
The affected components of the community should
vary in a like manner.
An even  bigger implication is that the surrogate
species concept is invalid in such a situation. As one
examines the community data in this report and in the
other reports (Mount et al.,  1984; Mount, Steen and
Norberg-King, 1985), it is clear that there is no set
response pattern of the community. Sometimes the
benthic invertebrates and the periphyton have similar
responses and  both are different from  the fish.
Sometimes the  fish and periphyton have  similar
responses and these are  unlike the benthic inverte-
brates.
The same is true of the test species. Sometimes the
Ceriodaphnia  respond like the periphyton and  other
times like the fish. The  important point is that a
careful  analyses of our knowledge of toxicology,
effluent decay, and relative sensitivity tells us that we
cannot expect:

  1.   Ceriodaphnia toxicity to always resemble toxicity
      to benthic invertebrates

  2.   Fathead  minnow toxicity to always resemble
      toxicity to fish
  3.   Fathead minnows and Ceriodaphnia to resemble
      each other in sensitivity or to display the same
      relative sensitivity to different effluents.

Any test species should have a sensitivity represent-
ative of some components of the community. The
important distinction is that one never can be sure
which components they will represent.

In comparing toxicity test results to community
response, comparison must be made with the  above
in mind. Certainly those community components that
are most sensitive will be most impacted and/or lost.
The  response of the  most sensitive test  species
should, therefore, be used to compare to the response
of the most sensitive of the community.
 A weakness in using the number of species as the
measure of community response is that species may
be severely affected yet not be absent. The density of
various species is greatly influenced by competition
for available  habitat, predation, grazing, and/or
secondary effects which may result from changing
species  composition. Density  is more  subject  to
confounding causes, other than direct toxicity, and is
not as useful as the species richness in the com-
munity to compare community response to measured
toxicity.
Several  measures of community structure are based
on number of species, e.g., diversity and community
loss index. Since diversity measures are little affected
by changes in the number of species  (or taxa) that are
in very low densities in the community, diversity is an
insensitive measure for some  perturbations which
can be measured by toxicity tests.  The  community
loss index is based only on the presence or absence of
specific species relative to a reference  station and
would be  useful except that habitat  differences
between stations heavily effect this measure. There
are several problems when using the number of (taxa)
species  measured. The  foremost is that the  mere
presence  or absence of species is not a compre-
hensive indicator of community health, especially if
the species are ecologically unimportant. Secondly, a
toxic stress may not eliminate species but yet have a
severe effect on density; presence or absence does
not consider such partial reductions. The presence or
absence of species as  the measure of community
impact is influenced by the chance occurrence of one
or a few individuals due to either drift, immigration, or
some catastrophic event when  in fact that species is
 not actually a part of the community where it is found.
 Effects  other than  toxicity,  such  as habitat, will
always confuse such comparisons to toxicity data to
some extent. They cannot be eliminated.

 The on  site  testing period  of this study began at
 extremely high flows as a result of rains in the upper
 basin. The apparent oxygen demand of the river water
 and the fungus problem are discussed in Chapter 4.
 The flows diminished greatly after the testing period
 (Table 4-11)  and a decision was made to re-run the
 ambient station toxicity tests.  Resources prohibited
 another on site study in which a new sample could be
 used each day. Instead, a sample was collected and
 shipped to ERL-D and was  used to renew  the test
 solution daily. This approach is totally dependent on
 one grab sample being representative whereas  the
 usual procedure of using seven different samples for
 the 7-day exposure period makes any one sample less
 important.
 For comparisons of the  toxicity prediction to  the
 community response, only the September data have
 been used because those samples were collected at a
 flow much more representative of the flows that
                        8-2

-------
 prevailed during most of the period  in which the
 substrates  were in the river and  when  the zoo-
 plankton was sampled. Since the toxicity tests were
 done on one grab sample taken at the same time as
 the zooplankton samples,  conditions should  have
 been  more  similar for  zooplankton  than for the
 macroinvertebrates which  were responding to the
 preceding four weeks of exposure. For acute effects
 such as spills or short high concentration exposures,
 the zooplankton and the toxicity tests should be most
 closely similar. For  chronic effects requiring several
 days to be manifested, the similarity might be much
 less.


 8.1   Effluent Tests
 Due to the large flows involved and the number of
 discharges,  dye  dilution measurements were  not
 economically possible. Therefore, the study  design
 was based on a comparison of ambient tests to field
 biological data and the effluent dilution tests were not
 necessary to the comparison. Effluents were tested to
 provide general-type information and  to locate any
 unusually high sources of toxicity.  None  of  the
 effluents had instream waste concentrations (IWC)
 after mixing that were greater than the acceptable
 effluent concentration  (AEC). Since  AEC's  were
 measured using water from just upstream of each
 effluent, the dilution waters contained all IWC's of
 upstream effluents and any effluent interaction (e.g.,
 additivity or antagonism) present is incorporated into
 the measurement. An exception to  this generality
 occurs wherever several effluents were diluted with
 the same water. In these cases all IWC's of upstream
 effluents were not in the dilution water and additive
 effects if present would not be measured. An example
 would be the 5 effluents diluted with Station 90.4
 water.
8.2   Comparison of Ambient Toxicity to
Biological Response
Table 8-1 contains a summary of the number of taxa,
young per female for the Ceriodaphnia and weights
for the fathead minnows for the field and toxicity data.
The  highest value for  each data  set was used to
calculate the percent reduction for all other values in
the set. The highest percent toxicity and the largest
percent reduction in number of taxa were then used
to develop Table 8-2 which  gives the percent of
correctly predicted responses using various arbitrary
levels  of impact/toxicity. Because there were few
impact/toxicity values above 60 percent, the percent
of correctly predicted stations is high when 60-80 or
80-100 percent levels are compared because these
are all  no-effect comparisons. The 20-40 percent
toxicity level gives approximately 60 percent correct
predictions for 20-40 and 40-60 percent levels of the
field data. None of the toxicity values were signif-
 icantly different from each other, and all were less
 than 40 percent below the highest value suggesting
 that any toxicity if present was slight. Anyone level of
 percent impairment is not  being proposed as  the
 correct percentage at this time.  This study is  not
 sufficient to judge which impairment of instream
 biological response data will correspond to a specified
 level of laboratory toxicity. Similar comparisons for all
 eight study sites (see Foreword) need to be completed
 before making decisions or recommendations.

 One should expect a general but not a point-by-point
 correlation between amount of toxicity and number of
 taxa lost. This expectation  is not due to error in
 measurement of toxicity  of taxa or  experimental
 variation,  but is expected because of the different
 relative sensitivity of test and community species.
 Added on top of this  variability are the confounding
 effects of measurement error. In addition, there is the
 chance collection of a few individuals of a species that
 does not  usually occur in that location and these
 numbers bias the number of taxa found. Events such
 as toxic spills before the  study period could  have
 residual effects on the community which would not
 be measured by the  toxicity tests.  General water
 quality conditions and physical effects, nontoxic in
 nature, such  as low DO, high temperature, or direct
 activities of man (like gravel removal or dredging) also
 might have affected  the community in the period
 preceding the study but would not affect the toxicity
 values.

 As discussed by Mount et al. (1985), point-by-point
 statistical comparisons, such as analyses of variance,
 may not show significant  differences even though
 definite trends are evident. Figure 5-1 is a plot of the
 number of taxa vs. river kilometers. There is definitely
 a decrease in the number of taxa of macroinverte-
 brates from upstream to downstream. The number of
zooplankton appear to be  lower in the upper  river
down to river kilometer 88.0. The two groups do not
reflect the same trends.

Some  amount  of  the change from upstream  to
downstream might be attributed to changing stream
flow or tributary recruitment areas. Gradient would
not be involved because the entire study reach was in
three pools  formed   by navigation  dams. Sharp
increases in numbers of taxa as shown from RK 88 to
68, all of which is in one pool, would suggest other
causes such as water quality.

Figure  8-1 is a  plot  of  the  percent toxicity for
Ceriodaphnia  and  the percent reduction in  zoo-
plankton taxa for each station. The correlation (r =
0.728) between these values suggess that the trends
in taxa are due to toxicity of the water. There certainly
was  addition  of  organic  matter from the  many
dischargers, including POTW's and this could have
enriched the water enough to produce the increase in
                                                                       8-3

-------
Table 8-1.    Number of Taxa, Number of Ceriodaphnia Young per Female and Fathead Minnow Weights with the Associated
              Percent Reduction Using the Highest Value of Each as Zero Percent at Various Stream Station (RIC), Kanawha River
Number of
Zoo-
River plankton
Kilometer Taxa
20.1
25.7
51.8
61.3
66.0L
66R
68.4
71.1
76.1 L
76.1 R
80.5
83.5
84.5
87.4L
87.4R
88.0L
88.0R
90.4
92.5L
92.5R
94.1
99.1
101.4
105.2
108.9
112.0
114.2
118.4
119.1
125.5
133.2
139L
139R
142.7
145.0
7
7
10
10
9
10
10
7
7
7
7
„
7
5
3
5
4
4
5
5
4
4
4
5
8
7
5
5

7
6
7
4
5
3
Percent
Reduction
30
30
0
0
10
0
0
30
30
30
30
	
30
50
70
50
60
60
50
50
60
60
60
50
20
30
50
50

30
40
30
60
50
70
Number of
Macroin-
vertebrate
Taxa
.24
21
20
15
16
19
20
24
24
21
19
._
21
14
24
16
21
22
23
23
20
32
—
21
26
22
25
—
27
28
30
27
27
28
28
Percent
Impact
25
34
37
53
50
41
37
25
25
34
41
—
34
56
25
50
34
31
28
28
37
0
—
34
19
31
22
—
16
12
6
16
16
12
12
Highest
Taxa
Impact
30
34
37
53
50
41
37
30
30
34
41
,
34
56
70
50
60
60
50
50
60
60
60
50
20
31
50
. 50
16
30
40
30
60
50
70
Mean
Number of
Young per
Cerio-
daphnia
28.8
24.8
31.8
32.3
--
31.4
29.0
--
27.4
30.4
25.0

25.9
23.6
26.9
27.3
24.1
23.1
27.6
24.8
26.4
24.0
26.9
24.7
26.8
26.8
—
--
27.6
28.0
24.8
24.3
21.5
24.7
22.3
Percent
Toxicity
11
23
2
0
--
3
10
--
23
6
23
--
20
27
17
15
25
28
15
23
18
26
17
24
17
17
--
--
15
13
23
25
33
24
31
Wt. of
Fathead
Minnows
.614
.619
.596
.577
--
.539
.605
--
.567
—
--
--
.565
.648
--
.637
--
.534
.620
--
.649
--
.616
--
.563
.552
—
--
.649
.597
.590
.560
.571
.608
.581
Percent
Toxicity
5
5
8
11
--
17
7
—
13
--
--
--
13
0
--
2
—
18
4
--
0
--
5
--
13
15
--
--
0
8
9
14
12
6
21
Highest
Percent
Toxicity
11
23
8
11
—
17
10
--
23
6
23
--
20
27
17
15
25
28
15
23
18
26
17
24
25
17
--
--
15
13
23
25
33
24
31
 Note: -- No data
                                                             Figure 8-1.    Percent toxicity to Ceriodaphnia vs. percent
                                                                           reduction of zooplankton taxa (Source Table
                                                                           8-1).
 Table B-2.    Percent of Stations Where Reduction in Number
               of  Taxa was  Correctly Predicted  by Toxicity
               Tests Using Four Arbitrary Levels of Comparison


                        Percent Reduction in Taxa
Percent
Increase in
Toxicity
20-40
40-60
60-80
80-100


20-40
57
3
3
3


40-60
60
39
42
42


60-80
53
77
81
81


80-100
45
100
100
100
                                                               40 r
£• 30
                                                             £20
                                                             0
                                                               10
                                                                        10     20     30     40     50
                                                                               Percent Reduction in Taxa
                                                                                                           60
                                                                                                                  70
                             8-4

-------
 zooplankton. However, one would expect enrichment
 to increase density, more than the number of taxa.
 Evidence for this effect is seen in that the downstream
 stations had the  highest density of zooplankton by
 several times, but the number of taxa is 30 percent
 lower between the first and second two.

An examination of Table 8-1 will  show  that the
reduction in taxa in the lower river was greater for the
macroinvertebrates and in the upper river, reductions
were greater for  the  zooplankton. The number of
macroinvertebrates taxa was greatest at RK 99.1.
Upstream of that Station, only two of the 11 values
were less than 24, while downstream of RK 99.1 all
20 values were 24 or less which is indicative of the
reduced taxa in the lower river.

Figure 8-2 is a plot showing the young per female
from  the ambient toxicity test and the number of
zooplankton taxa  plotted  against river kilometer.
(Recall that the samples for the September ambient
toxicity tests and  the zooplankton collections were
obtained at the same time (Figure  8-2).  This  plot
shows an amazingly similar pattern for both number
of taxa and young per female not evident in Figure
8-1. Correlation of percent toxicity  of the Cerio-
daphnia and percent  reduction of zooplankton was
highly significant(P<0.005%). Since ample food was
fed in the ambient toxicity tests to provide for at least
20-25 young per  female (the expected number of
young per female  we obtain in sterile reconstituted
water containing no food) nutrient enrichment would
not explain the pattern obtained. The pattern would
not be  up and  down  but should rather show a
continual increase in  number  of young  in  the
downstream direction. The peak around RK 120 and
the depression around RK 90 are not in concert with
the nutrient hypothesis. These data provide evidence
that the ambient tests are reflecting  the effect of
water quality on the instream zooplankton popula-
tions. And bearing in mind discussion earlier in this
section, i.e., that one does not expect a  tight correla-
tion between degree  of  toxicity  and amount of
community impact, the obvious correlation shown in
Figure 8-2 is also evidence that the ambient tests are
reflecting a response of the community to water
quality.  However, neither test species  reflected the
response of the macroinvertebrates.

There were many more dischargers present in the
study reach that were not tested than  were tested.
None of the effluents tested could account  for the
ambient toxicity observed.  Additivity is not a viable
explanation for the difference because the toxicity of
the effluents  was  measured with all   upstream
effluents present in the dilution water except as noted
in section 8-1.

There are several probable reasons for the ambient
toxicity observed.
 1.   Some of the effluents not tested may be more
     toxic than those tested.

 2.   Due to the rainfall, the effluent toxicity meas-
     ured was not representative either because the
     effluents were not typical or  their  measured
Figure 8-2.   Number of young per female Ceriodaphnia and number of zooplankton taxa at various river stations.

                       	0	o                                    ° Mean number of young
                   ,-•—"""        ^^"^       o                            per Ceriodaphnia
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River Kilometer
                                                                      8-5

-------
    toxicity was  not typical because the dilution
    water as a result of the flood condition gave a
    different  response (more suspended solids,
    BOD, etc.).
 3.  There are episodic occurrences of toxicity or
    other unknown sources that were missed in the
    effluent sampling  but which affected the in-
    stream community and were in some  of the
    September grab samples on which the ambient
    tests were run.


8.3.   Summary
The agreement between the ambient test data and
the community response was around 60 percent
using 20-40 and 40-60 percent levels for comparison.
In other studies in this series, the percent of correctly
predicted stations has  been generally higher. The
correlation between percent toxicity for Ceriodaphnia
and percent reduction of zooplankton taxa is highly
significant (P < 0.005%). The remarkable similarity
(Figure 8-2) between young per female and number of
zooplankton taxa is convincing data that the ambient
test measures effects  of water quality which  are
reflected in the community composition. The toxicity
tests did not correctly predict the macroinvertebrate
response supporting the need for multiple test species
and for including various groups in any biological
survey to identify impact.
                        8-6

-------
                                          References
Hamilton, M.A.  1984.  Statistical  Analysis of the
  Seven-Day Ceriodaphnia reticu/ata Reproductivity
  Toxicity Test.  EPA Contract J3905NASX-1.  16
  January. 48 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. and T.J. Norberg. 1985. Validity of Effluent
  and Ambient Toxicity for Predicting Biological
  Impact on Scippo Creek, Circleville,  Ohio. EPA
  Research Series, EPA-600/3-85-044.
Mount, D.I., A.E. Steen, and T.J. Norberg-King, Eds.
  1985. Validity of Effluent and Ambient Toxicity for
  Predicting Biological Impact on Five Mile Creek,
  Birmingham, Alabama. EPA/600/3-85/071.
Mount, D.I., N.A. Thomas, T.J. Norberg, M.T. Barbour,
  T.H. Roush, and W.F. Brandes.1984. Effluent and
  Ambient Toxicity Testing and Instream Community
  Response on the Ottawa  River, Lima, Ohio. EPA
  Research Series,  EPA-600/3-84-084.
Norberg, T.J. and D.I. Mount. 1985. A New Fathead
  Minnow (Pimephales prome/as) Subchronic Toxic-
  ity Test. Environ. Toxicol. Chem. 4(5):711 -718.
Rogers, J. 1984. University of Wisconsin at Superior,
  Wisconsin, and EPA Environmental Research Lab-
  oratory at Duluth, Minnesota. July.  Personal com-
  munication.
Sokal, R.R. and  F.J. Rohlf.  1981. Biometry.  W.H.
  Freeman and Company, New York.
Steele, G.R. and J.H. Torrie. 1960. Principles and
  Procedures of Statistics, a  Bio-Metrical Approach.
  2nd Edition. McGraw-Hill,  New York, 633 pp.
Weber, C.I. 1973. Recent Developments in the Meas-
  urement of the Response of Plankton and  Peri-
  phyton to Changes in Their Environment, in Bio-
  assay  Techniques and Environmental Chemistry
  (G.E. Glass, Ed.), pp. 119-138. Ann Arbor Sci. Publ.,
  Ann Arbor, Mich.
                                              R-1

-------

-------
                                         Appendix A
                           Toxicity Test and Analytical Methods
The Kanawha River study was  conducted in two
parts. One set of tests was conducted 14-21 August
1984.  Because the  river  stage  was very high,  a
second set of ambient tests was done on one set of
shipped samples  19-26 September  1984. All tests
were performed in a mobile laboratory either on site
or on shipped samples at the Environmental Research
Laboratory-Duluth, Minnesota.


A.1   On Site Test Methodology
The effluent samples were 24-hour composite sam-
ples collected using automatic samplers. The ambient
samples were grab samples taken daily for  seven
days. All samples were put in collapsible polyethylene
containers, with a capacity of either 1 or 5 gallons.
Composite samples were  terminated before 1200
hours on each day. The specif ic time was different for
each effluent. All ambient samples  were collected
between 0700 and 1400 and were collected close to
the artificial substrates.

As the samples were delivered to the mobile lab, they
were warmed to 25 °C, and then stacked in an air-
conditioned room  until used. Effluent dilutions were
made using polypropylene-graduated cylinders and
polyethylene beakers for mixing. All river water was
strained through  a fine-mesh screen  to  remove
zooplankton. A 2,000-ml volume of each was made;
200 ml were used for Ceriodaphnia tests and the rest
for  the fathead minnow tests. Initial DO, pH, and
conductivity measurements were taken before the
sample was split. Dedicated polyethylene containers
were used  for  each  concentration  for both the
Ceriodaphnia and the fathead minnowtests. Effluents
were diluted with water upstream of each outfall or
group of outfalls, and these stations are identified on
Table 3-2.

As the  ambient samples were collected, they were
put in two  sets of dedicated polyethylene containers
for  the  Ceriodaphnia and fathead minnow  tests.
Ambient stations were close together, the flow-time
between stations was short, and the stations used for
dilution water for effluents were scattered among the
rest of the  stations. Therefore, the initial DO, pH, and
conductivity measurements were done only on the
dilution water stations and not on the rest of the
stations in  order to reduce work load. Final DOandpH
measurements were taken for all  stations. For both
test species, a new sample of effluent or ambient
water was used for each daily change.

The Ceriodaphnia test followed generally the pro-
cedures of Mount and Norberg (1984). Adult Cerio-
daphnia were transferred to the dilution water two
days prior to the initiation of the first tests  on site. A
young  Ceriodaphnia (0-6 hour old) was placed in a
1 -oz plastic portion cup in 15 ml of test solution. There
were ten animals for each treatment. Each day the
animal was  removed with an eyedropper and placed
into a new cup containing  new test solution. When
young  were present,  they  were counted and dis-
carded. Each set of five effluent concentrations and
the dilution water control were randomly assigned to
a row on a test board.Each  test board held five test
organisms per concentration, and each test was split
into half-test boards. The ambient station samples
were run in the same manner,  with  six ambient
stations randomly arranged in rows on each half test
board. In this manner, treatments could be assigned
randomly and  independently to each half  tray. The
rotation and shelf assignment of each half tray was
randomized  each day.

A food suspension was fed daily after each change.
The food consisted of three parts: (1) 5 g/L of dry
yeast, (2) 5 g/L of Cerophyl®, stirred overnight and
filtered through a plankton net, and (3) 5 g/L of trout
chow,  aerated vigorously for 7 days,  settled, and
decanted. The yeast suspension and the supernatant
from the Cerophyl® and trout chow were  mixed in
equal parts,  and new food was made every 7 days.
The mixture, the Cerophyl®, and the yeast compo-
nents were refrigerated, while the trout chow super-
natant was frozen until the mixture was made. This
food is suitable for a  wide variety of water types,
including reconstituted water. This mixture  is fed 0.1
ml per day per Ceriodaphnia rather than 0.05 ml as
was recommended for yeast (Mount and Norberg
1984),  because  the suspended solids are around
1,800 mg/L, less than half the solids contained in the
yeast suspension.

The methods for the fathead minnow tests followed
closely those described by Norberg and Mount (1985).
The test chambers were 30.5 x 15.2 x 10.2 cm and
©Cerophyl was obtained from Agri-Tech, Kansas City, Missouri. As of
 January 1985, Cerophyl® was no longer being produced by that manufac-
 turer. Use of trade names does not constitute endorsement.
                                               A-1

-------
divided into four compartments; this design allowed
four replicates for each concentration.  The larval
fathead minnows were < 24 hours old and from the
ERL-Duluth culture. The fish were assigned to the
test compartments by pipetting one or two fish at a
time to each replicate testchamber until all replicates
had ten fish in each, or forty per concentration. All
treatments were re-randomized daily with respect to
position on the shelves. Newly hatched brine shrimp
were fed to the fish three times a day. The uneaten
shrimp were removed daily by siphoning the tanks
during test solution renewal. At the same time, the
volume in the test chamberwas drawn down to 1 cm,
after which 2 L of new test solution was added. The
laboratory temperature  was  25±1°C. A 16-hour
light photoperiod was used. Because DO was low at
the end  of the first 24  hours, test volumes were
reduced to 1 L After 7 days of exposure, the fish were
preserved in 4 percent formalin. Prior to weighing,
they were rinsed in distilled water. Then  each group
was oven dried for 18 hours  in pre-weighed alumi-
num pans and weighed on  a five-place analytical
balance.
station data, Tukey's Honestly Significant Difference
Test (Sokal and Rohlf, 1981) is used for the ambient
toxicity test data to compare stations.


A.3.2  Fathead Minnows
The four groups' mean weights are analyzed statis-
tically with the assumption that the four test-chamber
compartments behave as  replicates. The method of
analysis assumes the variability in the mean treat-
ment response is proportional to the number of fish
per treatment.  MINITAB  (copyright, Pennsylvania
State  University 1 982)  was  used to estimate a t-
statistic for comparing the  mean  treatment and
control data using weighted regressions with weights
equal to the number of  measurements in the treat-
ments.

The t-statistic was then compared to the critical t-
statistic for the standard  two-tailed  Dunnett's test
(Steel  and Torrie  1960). The  survival  data were
arcsine-transformed prior  to the regression analyses
to stabilize variances for percent data.
A.2   Methods for Shipped Samples
Only ambient samples were tested in the September
testing on shipped samples. Five-gallon grab samples
were collected on 15 September 1984, cooled using
wet ice, and shipped to Duluth and stored at 6° C.
Each day, an  aliquot was warmed to 25° C, and the
new aliquot was exchanged in the Ceriodaphnia and
fathead minnow tests. Otherwise, the same equip-
ment and procedures were used as described for the
onsite tests.

A.3   Statistical Analyses

A.3.1   Ceriodaphnia dub/a
The  statistical analyses were performed  using the
procedure of Hamilton (1984) as modified by  Rogers
(1984). In this procedure, the young production data
were analyzed to obtain the mean number of young
per female per treatment. Daily means were calcu-
lated and 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 the overall toxicity
effects. Confidence intervals are calculated by the
bootstrap procedure. This 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
                       A-2

-------
                                         Appendix B
                       Biological Samplng and Analytical Methods
B.I   Periphyton Survey
The periphytic community was sampled quantita-
tively using clear acetate strips suspended  in the
Kanawha River at the same locations as the artificial
substrates for the benthic macroinvertebrates (Table
3-2). Triplicate strips were placed in the river at the 33
stations on  14-20 August, 1984 and retrieved on
12-17 September, 1984. The strips were preserved in
formalin until analysis. The strips were scraped and
the material was analyzed for chlorophyll  a  and
biomass (ash-free dry weight, AFDW).

For AFDW, samples 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
before final weighing, and biomass was expressed in
g/m2. Filters for chlorophyll a analysis were macer-
ated in a 90 percent acetone solution, then centri-
f uged and analyzed spectrophotometrically. A chloro-
phyll a standard (Sigma Chemicals) extracted in a 90
percent acetone  solution was used for instrument
calibration. Chlorophyll a standing crop was ex-
pressed as mg/2. The biomass and chlorophyll a data
were used to calculate the Autotrophic Index (Weber,
1973),  which indicates the relative proportion of
heterotrophic and autotrophic components  in the
periphyton.
B.3  Macroinvertebrate Methods
Hester-Dendy samplers (round plate, variable spaced,
about 0.1  m2)  were suspended  in the river at  33
locations (Table 3-2). The samplers were set from
August 14-20, 1984 and were removed from Sep-
tember 12-17,  1984.  The goal  was  to  have the
samplers from 2-3 feet from the normal pool surface.
Because  they  were  set  during high water, the
positioning was done by measuring water depth and
then calculating the  depth from the bottom that
should be selected at the existing river stage.

The samplers were retrieved by raising them to just
under the surface and then a net was placed beneath
them and they were lifted out. The entire sampler and
contents was preserved in 10% formalin containing
rose bengal stain.

For enumeration, the plates were scraped with a
putty knife to remove all material. This material was
then washed to remove silt and then strained through
a 500/u mesh netting. The organisms were picked
from the debris under 8X magnification and placed in
70% alcohol. Identification was to the lowest taxon
within the expertise of the analyst.
B.2  Zooplankton Methods
Zooplankton were collected from thirty-three stations
on the Kanawha River in West Virginia on 15,16, and
17  September,  1984. Samples  were collected in
triplicate at each station, at 3-foot depths by pumping
200 liters of water through a 153 //m mesh  net.

In the laboratory, the samples were concentrated by
allowing the contents of the  sample container to
settle, and siphoning from the top as much  liquid as
possible without disturbing the plankton. The entire
sample was enumerated by placing  approximately
5-ml at a time on a Ward zooplankton counting wheel
and identifying to the lowest possible taxon. Identi-
fications were made using a dissecting scope at 25X
magnification, and those organisms which could not
be identified at that power were mounted and viewed
under a compound scope at a higher magnification.
                                              B-1

-------

-------
                                             Appendix C
                                          Tox/city Test Data
Table C-1.    Water Chemistry Data for Effluent Toxicity Tests. Values are for Both Ceriodaphnia and Fathead Minnow Tests and
            Final Dissolved Oxygen Values are for Daphnids Only
Effluent (RK)a
POTW1I
(90.4)





POTW1M
(90.4)





POTW2
(89.6)





POTW3
(66.8)





A(143.5)






8(111.0)






C (1 1 1 .0)






Percent
Effluent
(v/v)
100
30
10
3
1
Dilution Water
(90.4)
100
30
10
3
1
Dilution Water
(90.4)
100
30
10
3
1
Dilution Water
(9O.4)
100
30
10
3
1
Dilution Water
(68.4)
100
30
10
3
1
Dilution Water
(145.0)
100
30
10
3
1
Dilution Water
(111.0)
100
30
10
3
1
Dilution Water
(114.2)
pH Range
6.5-7.1
6.9A"
6.6A"
6. 8 A"
6.8A"
6.5-7.0

6.2-6.5
6.6A"
6.7Ab
6.8A"
6.8A"
6.5-7.0

6.7-7.0
6.7Ab
6.5A"
6.5A"
6.5Ab
6.5-7.0

6.6-7.0
6.7A"
6.6Ab
6.6A"
6.6A"
6.0A"

5.4-7.8
6.7A"
6.6A"
6.5A"
6.5Ab
6.5-6.8

6.7-7.0
6.8Ab
6.8A"
6.7Ab
6.7A"
6.8

6.9-7.0
6.9Ab
6.9-7.0
6.9A"
6.8Ab
6.87.0


Mean
7.4
6.9
7.2
7.4
7.3
7.9

7.2
7.2
7.3
7.3
7.3
7.9

6.4
7.4
7.5
7.4
7.3
7.9

6.1
7.0
7.0
7.1
7.0
7.0

8.6
8.0
7.9
7.9
7.9
7.5

8.2
7.3
7.2
7.3
7.3
7.5

4.3
7.3
7.3
7.3
7.2
7.5

Initial DO
(mg/L)
Range
5.7-8.6
-.
	
__
'
7.3-8.8

6.3-7.8
	
__
	
--
7.4-8.4

2.6-7.9
..
__
1
-_
7.1-8.8

5.7-6.4
	
,
__
	
—

7.9-9.2
1
—
__
_.
7.3-7.6

7.0-8.8
	
,
-
--
7.2-7.9

1.7-7.3
	
	
__
.-
7.2-7.9


Mean
7.4
7.0
7.3
6.7
7.3
7.0

5.6
6.4
6.5
7.0
6.9
6.8

6.2
7.0
6.3
7.3
7.3
7.2

6.4
6.3
6.4
6.6
6.4
6.4

7.3
7.4
7.4
7.3
7.3
6.8

6.9
6.9
6.9
6.8
6.8
6.9

6.8
__
7.4
7.2
6.2
6.9

Final DO
(mg/L)
Range
5.6-7.7
5.4-8.2
5.9-7.9
5.5-7.7
6.3-7.7
6.3-7.6

5.5-5.7
5.4-7.0
5.1-7.5
5.8-7.8
6.2-7.8
6.3-7.8

5.5-6.8
6.2-7.8
4.9-7.7
6.5-8.0
6.4-7.9
6.5-7.6

5.7-7.0
5.8-7.2
5.1-7.2
6.1-7.3
5.2-7.4
4.9-7.0

6.3-7.9
6.3-7.9
6.3-7.9
6.2-8.1
6.4-8.0
5.5-7.4

4.7-8.1
4.2-8.1
3.8-8.0
3.7-8.1
4.0-7.9
5.5-7.9

„
__
6.4-8.1
6.3-8.1
3.2-8.0
4.1 -8.0

Conductivity
(umhos)
3,075




111

446




111

480




111

545




140

162




93

132




90

1 5,083




90

                                                  C-1

-------
Table C-1.
Effluent
0(112,2)





E(88,2)






F (85,8)






G (85.8)






H (85.8)






1 (77.4)






J (68.9)





K (67.1 )






(continued)
Percent
Effluent
(v/v)
100
30
10
3
1
Dilution Water
(114.2)
100
30
10
3
1
Dilution Water
(90.4)
100
30
10
3
1
Dilution Water
(90.4)
100
30
10
3
1
Dilution Water
(86.1)
100
30
10
3
1
Dilution Water
(86.1)
100
30
10
3
1
Dilution Water
(80.5)
100
30
10
3
1
Dilution Water
(68.4)
100
30
10
3
1
Dilution Water
(68.4)
pH Range
7.0-7.2
6.9A"
6.9A"
6.8A"
6.8Ab
6.8

7.0-7.8
6.8A"
6.5A"
6.5A"
6.6Ab
6.7-7.0

6.4-6.8
6.7A"
6.7Ab
6.7Ab
6.6Ab
6.5-7.0

6.7-7.1
6.7A"
6.6A"
6.7Ab
6.6A"
6.7-6.9

7.0-7.4
6.7Ab
6.6A"
6.5Ab
6.6A"
6.6-6.9

7.0-7.2
6.8A"
6.6Ab
6.5Ab
6.6A"
6.5-6.6

6.3-7.0
6.7A"
6.8A"
6.8A"
6.7A"
6.8-7.2

6.9-7.3
6.8A"
6.8A"
6.6A"
6.6A"
6.6-7.2


Mean
8.3
7.4
7.3
7.3
7.3
7.5

7.9
7.8
7.8
7.9
7.8
7.4
1
11.8
8.8
8.1
8.0
7.6
7.3

6.8
7.6
7.6
7.5
7.4
7.1

8.2
7.4
7.5
7.4
7.3
7.1
'
7.1
7.5
7.4
7.2
7.2
7.2

6.4
7.0
7.2
7.2
7.0
7.0

7.5
7.2
7.1
7.2
7.1
7.0

Initial DO
(mg/L)
Range
7.5-8.9
—
—
..
7.2-7.9

7.5-8.4
--
—
—
-.
7.0-7.8

10.4-14.2
—
—
—
-.
7.0-7.6

4.6-8.0
--
—
—
—
6.9-7.4

7.2-9.0
—
-.
-.
--
6.9-7.3

6.1 -7.8
.-
—
.-
—
7.1-7.2

4.5-7.1
—
—
-_
6.9-7.0

6.6-8.
--
—
—
_-
7.0


Mean
7.1
6.5
6.7
6.8
6.8
7.0

6.7
8.2
8.2
7.3
7.4
6.9

7.6
7.8
7.3
7.3
7.2
6.8

6.9
6.1
6.8
7.0
7.2
7.2

6.7
--
7.1
6.7
7.3
7.1

6.9
7.3
7.1
7.2
7.2
7.2

6.1
5.7
6.1
7.0
7.1
7.2

7.0
--
6.7
6.4
7.3
7.3

Final DO
(mg/L)
Range
5.2-8.2
4.0-8.2
3.7-8.1
4.7-8.1
5.3-7.8
6.4-7.8

-
--
--
6.5-8.2
6.4-8.2
6.0-8.1

-.
6.6-8.2
6.4-8.1
6.5-8.1
6.0-8.0
6.0-7.9

6.5-7.3
5.1-7.8
5.7-7.8
5.5-7.8
6.2-7.8
6.0-8.0

-
--
—
4.5-8.1
6.4-8.0
6.3-7.9

6.4-7.3
6.5-7.9
6.4-8.0
6.3-8.0
6.5-7.8
6.7-8.1

5.5-6.7
5.4-6.3
4.0-7.2
6.2-8.0
5.9-8.0
6.5-8.0


—
5.9-7.8
5.2-7.3
6.5-8.1
6.6-8.1

Conductivity
(umhos)
175



90

1,250




90

14,083




90

242




90

591




90

1,017




95

1,275



140

132




140

•RK of the discharger, see Tables 3-1, 3-2.
bOnly one measurement was made.

                           C-2

-------
Table C-2.    Final Dissolved  Oxygen Concentrations for
              Fathead Minnow  Larval Growth Tests on
              Effluents, Charleston, West Virginia, August
              1984
Table C-3.    Initial Water Chemistry Data for Ambient Toxicity
             Tests with Ceriodaphnia and Fathead Minnows
             on Day 1 of Testing, Charleston, West Virginia,
             August 1984
Effluent
POTW11
(90.4)





POTW1M
(90.4)





POTW2
(89.6)





A(143.5)






Percent
Effluent
(v/v)
100
30
10
3
1
Dilution Water
(90.4)
100
30
10
3
1
Dilution Water
(90.4)
100
30
10
3
1
Dilution Water
(90.4)
100
30
10
3
1
Dilution Water
(145.0)
DO
Mean
1.8
2.5
2.4
3.0
3.1
3.2

2.8
3.1
3.1
3.3
3.6
3.4

1.7
2.7
2.9
3.1
3.1
3.5

3.6
3.9
3.5
3.1
3.8
3.4

(mg/L)
Range
1.6-3.8
1.2-4.6
1.5-4.2
2.3-5.2
2.8-5.2
2.6-4.9

0.5-4.5
1 .8-4.5
1.3-5.2
1 .2-5.4
1.2-5.9
2.0-4.6

0.7-4.5
1.4-7.6
1.9-6.2
2.4-5.1
2.1 -4.6
2.8-5.4

2.8-5.0
2.7-6.4
2.9-5.1
1.6-5.7
3.1-5.6
2.4-5.7

Note: Initial routine chemistry values are in Table C-1.
Ambient
Station (RK)
145.0
142.7
1 39.0R
1 39.0L
133.2
125.5
119.1
114.2
1 1 2.0
108.9
105.2
101.4
99.1
94.1
92.5R
90.4
88.0R
88.0L
87.4R
87.4L
Davis Creek, 85.8
83.3
84.5
84.5
80.5
76.1 R
76. 1L
71.1
68.4
66.0R
66.0L
61.3
51.8
25.7
20.1
pH
6.8
6.9
6.8
7.0
6.9
6.7
7.0
6.8
6.9
6.8
6.9
6.8
6.9
6.8
6.9
6.9
6.9
6.9
6.9
6.9
7.0
7.0
7.0
7.0
6.8
6.9
6.9
6.9
6.9
6.7
6.9
6.8
6.8
6.8
6.8
Initial DO
(mg/L)
7.9
8.0
8.1
8.0
8.1 '
8.3
8.0
8.0
8.0
8.1
8.1
8.1
7.8
8.0
7.7
8.0
8.1
8.0
8.0
7.8
7.9
8.1
8.1
8.1
8.0
8.0
8.2
8.1
8.0
8.3
8.0
8.1
8.1
8.1
.-• 8.3
Conductivity
(yumhos)
98
105
98
110
102
105
108
87
100
102
95
123
100
100
95
90
96
95
93
155
'900
100
100
168
78
87
98
110
121
100
108
100
105
96
98
                                                                                    C-3

-------
Tablo C-4.    Final Dissolved Oxygen Concentrations for
             Ambient Toxicity Tests with Ceriodaphnia and
             Fathead Minnow, Charleston,  West Virginia,
             August 1984
Ambient
Rfn^inn
(RK)

145.0
142.7
139.0R
139.0L
133.2
125.5
119.1
114.2
112.0
108.9
105.2
101.4
99.1
94.1
92.5R
90.4
88.0R
88.0L
87.4R
87.4L
Davis Creek, 85.8
83.3
84,5
80.5
76.1 R
76.1 U
71.1
68.4
66.0R
66.0L
61.3
51.8
25.7
20.1
Ceriodaphnia
Final DO (mg/L)
Mean

6.6
6.9
7.2
7.0
6.9
6.9
7.3
6.4
7.4
6.7
6.9
7.3
6.1
7.0
6.7
6.8
7.2
6.25
7.3
7.0
6.9
7.1
6.7
6.9
7.4
7.0
6.9
6.8
6.1
6.4
6.7
6.9
6.5
6.2
Range

6.2-7.3
6.2-7.9
5.8-7.9
6.2-7.4
6.0-7.7
6.1-8.0
7.1-7.6
5.6-7.0
6.1-8.1
6.0-7.5
5.5-7.8
5.8-7.9
5.1-7.5
6.2-7.4
5.5-7.7
6.0-7.5
6.0-7.7
6.2-6.3
6.1-8.1
6.1-8.0
5.8-7.8
6.3-7.6
4.6-8.0
6.1-7.4
6.1-8.1
6.0-7.8
6.1-8.1
5.6-7.6
4.8-7.7
5.8-7.1
6.2-7.1
6.1-7.9
5.1-7.4
5.7-7.2
Fathead Minnow
Final DO (mg/L)
Mean

3.8
4.1
4.1
3.8
3.8
4.1
3.9
3.8
3.7
4.0
4.3
4.0
4.6
4.4
4.5
4.2
3.9
4.2
3.9
4.4
3.9
4.1
4.2
3.9
4.2
3.8
4.0
3.9
4.2
3.8
4.1
4.2
4.2
3.8
Range
t
2.6-5:5
1.9-5J5
3.0-5lO
7.5-4.7
2.8-4,7
3.1-4;9
2.2-5;4
2.3-6!1
2.6-4.7
3.5-4.9
3.3-5.1
3-4.7
3.5-5.9
3.2-5;6
3.4-5:6
2.8-5l2
3.1-5.4
3.1-5.1
2.2-5.0
3.0-515
2.4-4.8
2.7-5.4
3.2-5.3
2.8-4.9
3.0-5.4
2.4-4.9
2.7-5.2
2.1-5.5
2.9-5i6
2.8-4f7
3.1-5.1
2.9-5^6
2.9-5,3
2.4-4,5
                            C-4

-------
Table C-5. Water Chemistry Data for Ceriodaphnia and Fathead Minnow Ambient Toxicity Tests, Kanawha River, Charleston,
West Virginia, September 1 984
Fathead Minnow
Ambient
Station
(RK)
145.0
142.7
139.0R
139.0L
133.2
125.5
119.1
112.1
108.9
105.2
101.4
99.1
94.1
92.5R
92.5L
90.4
88.0R
88.01.
87.4L
87.4L
84.5
80.5
76.1 R
76. li-
es^
66.0
61.3
51.8
25.7
20.1


PH
7.1
7.0
7.0
7.0
6.9
6.8
6.8
6.9
6.8
7.0
7.0
7.0
7.0
7.0
7.0
7.0
7.0
7.2
7.0
7.0
6.9
—
7.0
6.8
6.9
6.8
6.8
6.9
6.8
7.0

Conductivity
(//mhos)
115
160
160
160
140
150
140
125
140
150
150
140
150
120
135
140
360
140
290
250
170
—
180
180
200
210
220
170
160
150
Initial
DO
(mg/L)
9.0
9.4
9.3
9.2
9.2
9.1
9.0
9.1
8.8
9.0
8.9
9.0
8.9
8.7
9.0
9.1
8.4
9.0
9.2
8.7
8.9
—
8.3
9.0
9.0
8.8
8.7
8.9
9.1
8.9


Mean
6.6
6.8
6.8
7.1
6.8
6.8
6.8
6.7
6.6
--
6.8
—
6.6
— •
7.0
6.8
--
6.7
.-
6.8
6.8
—
--
6.8
6.8
6.7
6.5
6.7
6.9-
6.5
Final DO
(mg/L)
Range
5.8-7.1
5.9-7.7
5.4-7.5
6.7-7.5
5.7-7.7
5.7-7.4
5.9-7.5
6.1-7.5
5.1-7.7
--
6.3-7.6
—
5.5-7.1
--
6.8-7.2
5.4-7.5
—
4.9-7.3
--
6.0-7.5
5.8-7.4
—
--
5.6-7.3
5.2-7.6
5.9-7.4
4.6-7.4
6.1-7.1
6.3-7.3
5.3-7.5
Ceriodaphnia


Mean
7.7
7.8
7.8
8.0
7.9
7.8
7.5
7.9
7.9
7.9
8.0
7.9
7.7
7.7
7.7
7.6
7.9
7.7
7.9
7.7
7.8
7.6
7.8
7.8
7.8
7.8
7.9
7.8
7.7
7.7
Final DO
(mg/L)
Range
7.6-7.8
7.8-7.9
7.4-8.1
7.7-8.2
7.7-8.1
7.8-7.9
6.8-7.8
7.8-7.9
7.8-8.1
7.8-8.0
7.6-8.3
7.8-7.9
7.6-7.9
6.7-8.5
7.4-7.9
7.0-7.9
7.8-8.1
6.7-8.5
7.7-8.1
7.4-7.8
7.6-8.0
7.3-7.9
6.9-8.5
7.8-9.9
7.6-7.9
7.6-7.9
7.8-8.1
7.6-8.1
6.6-8.4
7.5-8.1
C-5

-------

-------
                                               Appendix D
                                             Biological Data
Table D-1.    Routine Chemistry Data for August and September for the Stream Stations. Readings Were Taken When Artifical
             Substrates Were Set and Removed, Kanawha River
Station
(RK)
20.1
25.7
51.8
61.3
66.0L
66.0R
68.4
71.1
76.1 L
76.1 R
80.5
84.5
87.4
87.4
88.0R
88.0L
90.4
92.5L
92.5R
94.5
99.1
101.4
105.2
108.9
112.0
114.2
119.1
125.5
133.2
139.0L
139.0R
142.7
145.0
Conductivity
(//mhos)
Aug.
129
135
130
143
130
145
220
133
134
134
139
140
146
136
118
279
115
115
107
116
117
115
111
105
97
94
95
94
97
96
96
99
96
Sept.
157
153
198
223
203
204
197
184
177
180
187
167
220
196
139
310
148
150
101
152
149
154
148
173
169
171
180
180
170
123
127
134
110
DO
(mg/I)
Aug.
7.6
6.9
7.1
7.2
7.3
7.5
7.6
7.9
7.8
7.9
7.5
8.0
7.6
7.7
8.1
7.3
8.0
7.9
7.9
7.8
7.9
8.1
7.9
8.4
8.9
8.3
8.4
8.3
8.3
8.3
8.4
8.4
8.2
Sept.
7.4
7.0
6.7
6.8
7.0
7.2
6.7
7.1
7.5
7.8
7.4
7.5
7.9
7.9
7.7
7.8
7.7
7.8
7.8
7.8
7.7
7.8
8.0
7.2
7.2
7.4
8.7
8.4
8.0
8.8
9.1
7.8
7.8
PH
Aug.
7.0
7.3
7.2
7.2
7.3
7.3
7.4
7.3
7.3
7.4
7.3
7.2
7.3
7.5
7.4
7.4
7.3
7.4
7.3
7.3
7.4
7.4
7.2
7.4
7.4
7.4
7.3
7.2
7.4
7.4
7.5
7.6
7.6
Sept.
7.0
7.0
7.4
7.1
7.2
7.1
7.1
7.2
7.3
7.3
7.2
7.2
7.4
7.3
7.3
7.4
7.2
7.2
7.1
7.2
7.3
7.2
7.1
7.3
7.9
8.0
7.8
7.8
7.7
7.3
7.8
7.6
7.6
Temp.
(°C)
Aug.
22.6
22.5
23.2
23.6
24.3
24.8
24.7
25.2
25.0
24.8
24.7
24.5
24.1
24.5
23.8
26.0
22.2
22.2
21.9
22.3
22.4
22.3
22.2
22.3
24.4
22.8
22.7
22.1
22.1
21.6
21.5
21.7
21.8
Sept.
21.8
21.9
22.5
23.4
23.1
22.8
23.3
22.6
22.4
22.7
22.4
21.7
22.5
21.9
21.7
23.0
21.6
22.3
22.2
22.6
22.3
22.4
22.5
23.2
23.1
23.0
22.4
24.0
21.4
21.3
21.4
21.4
19.9
                                                    D-1.
                                                                           U. S. GOVERNMENT PRINTING OFFICE: 1986/646-116/40662

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