vvEPA
United States
Environmental Protection
Agency
Permits Division
(EN-336)
Washington DC 20460
Environmental Research
Laboratory
Duluth MN 55804
IP,
EPA/600/3-8fc/071
March 1986
Research and Development
Validity of Ambient
Toxicity Tests for
Predicting Biological
Impact, Ohio River,
Near Wheeling,
West Virginia
-------
EPA/600/3-85/071
March 1986
Validity of Ambient Toxicity
Tests for Predicting Biological Impact,
Ohio River, Near Wheeling, West Virginia
Edited by
Donald I. Mount, Ph.D.1
Alexis E. Steen2
Teresa Norberg-King1
1 Environmental Research Laboratory
U.S. Environmental Protection Agency
6201 Congdon Blvd.
Duluth, Minnesota 55804
2EA Engineering, Science, and Technology, Inc.
Hunt Valley/Loveton Center
1 5 Loveton Circle
Sparks, Maryland 21152
Environmental Research Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Duluth, MN 55804
-------
Disclaimer
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.
-------
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 effl uent 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 discharge to a receiving water.
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 in the
"whole effluent" approach which involves setting limits using effluent toxicity
as a control parameter. There are advantages and disadvantages to both
approaches.
The "whole effluent" approach eliminates the need to specify a limit for each of
thousands of substances that may be found in an effluent. It also includes all
interactions between constituents as well as biological availability. Such limits
determined on fresh effluent may not reflect toxicity of effluent after aging in the
stream and fate processes change effluent composition. This problem is less
important since permit limits are normally applied at the edge of the mixing zone
where aging has not yet occurred.
The following study site was on the Ohio River near Wheeling, West Virginia,
and was conducted in July and August 1984.
-------
To date, eight sites have been investigated involving municipal and industrial
discharges. They are, in order of investigation:
1. Scippo Creek, Circleville, Ohio
2. Ottawa River, Lima, Ohio
3. Five Mile Creek, Birmingham, Alabama
4. Skeleton Creek, Enid, Oklahoma
5. Naugatuck River, Waterbury, Connecticut
6. Back River, Baltimore Harbor, Maryland
7. Ohio River, Wheeling, West Virginia
8. Kanawha River, Charleston, West Virginia
This project is a research effort only and has not involved either NPDES permit
issuance or enforcement activities.
Rick Brandes
Permits Division
Nelson Thomas
ERL/Duluth
Project Officers
Complex Effluent Toxicity
Testing Program
-------
Contents
Page
Foreword iii
List of Figures vi
List of Tables vii
Acknowledgements ix
List of Contributors x
Executive Summary xi
Quality Assurance xii
1. Introduction 1-1
2. Study Design and Site Description 2-1
3. Ambient Toxicity Tests 3-1
3.1 River Flow Measurements 3-1
3.2 Chemical and Physical Test Conditions 3-1
3.3 Ambient Toxicity Test Results 3-2
3.4 Discussion 3-3
4. Plankton Community Survey 4-1
4.1 Community Structure 4-1
4.2 Evaluation of the Zooplankton Community 4-1
5. Periphyton Community Survey 5-1
5.1 Chlorophyll a and Biomass Measurements 5-1
5.2 Evaluation of the Periphytic Community 5-2
6. Macroinvertebrate Community Survey 6-1
6.1 Community Composition 6-1
6.2 Station Comparisons 6-2
6.3 Evaluation of the Macroinvertebrate Community 6-3
7. Comparison Between Laboratory Toxicity Tests and
Instream Biological Response 7-1
7.0 Background 7-1
7.1 Comparison of Toxicity Test Results and Field Data 7-2
References R-1
Appendix A: Toxicity Test and Analytical Methods A-1
Appendix B: Biological Sampling and Analytical Methods B-1
Appendix C: Additional Biological Data C-1
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List of Figures
Number Title Page
2-1 Study area on the Ohio River near Wheeling, West Virginia 2-1
4-1 Densities of crustaceans and rotifers collected in the Ohio River
near Wheeling, West Virginia, 1984 4-1
6-1 Mean density of oligochaetes in the Ohio River 6-2
6-2 Mean density of Gammarus amphipods in the Ohio River 6-2
6-3 Mean density of Chironomids in the Ohio River 6-3
6-4 Mean density of Trichopterans in the Ohio River 6-3
7-1 Percent toxicity and percent reduction in macroinvertebrate taxa
for eight ambient stations 7-4
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List of Tables
Number Title Page
3-1 Ohio River Flow (mVsec) 3-1
3-2 Water Chemistry Data for Ambient Toxicity Tests with Fathead
Minnows and Ceriodaphnia, Ohio River, Wheeling, West Virginia,
July 1984 3-2
3-3 Mean Survival of Larval Fathead Minnows for Ambient Toxicity
Tests, Ohio River, Wheeling, West Virginia, July 1984 3-3
3-4 Mean Individual Weights (mg) of Larval Fathead Minnows for
Ambient Toxicity Tests, Ohio River, Wheeling, West Virginia,
July 1 984 3-3
3-5 Mean Young Production and Percent Survival of Ceriodaphnia for
Ambient Toxicity Tests, Ohio River, Wheeling, West Virginia,
July 1984 3-3
4-1 Densities (No./liter) of Plankton Collected from the Ohio River,
Wheeling, West Virginia 1984 4-2
5-1 Chlorophyll a and Biomass Measurements of Periphyton Collected
from Artifical Substrates in the Ohio River, Wheeling,
West Virginia, August 1984 5-1
6-1 Mean Percent Composition of Major Macroinvertebrate Taxa,
Ohio River, Wheeling, West Virginia 6-1
7-1 Number of Macroinvertebrate Taxa Collected from the
Ohio River 7-3
7-2 Fathead Minnow Growth in Ambient Station Water 7-3
7-3 Ceriodaphnia Reproduction in Ambient Station Water 7-3
7-4 Percent of Stations Correctly Predicted Using Four Categories of
Percent Impact 7-4
C-1 Numbers of Plankton Collected from the Ohio River, Wheeling,
West Virginia, August 1984 C-1
C-2 Density (No./m2) and Percent Occurrence of Macroinvertebrates
Collected at Stations 1 and 2 in the Ohio River, Wheeling,
West Virginia, July-August 1984 C-2
C-3 Density (No./m2) and Percent Occurrence of Macroinvertebrates
Collected at Stations 3 and 4 in the Ohio River, Wheeling,
West Virginia, July-August 1984 C-4
-------
List of Tables (cont'd)
Number Title Page
C-4 Density (No./m2) and Percent Occurrence of Macro-invertebrates
Collected at Stations 5 and 6 in the Ohio River, Wheeling,
West Virginia, July-August 1984 C-5
C-5 Density (No,/m2) and Percent Occurrence of Macroinvertebrates
Collected at Stations 7 and 8 in the Ohio River, Wheeling,
West Virginia, July-August 1984 C-5
C-6 Numbers of Macroinvertebrates for Each Replicate Sample
Collected at Stations 1 and 2 in the Ohio River, Wheeling,
West Virginia, July-August 1984 C-8
C-7 Numbers of Macroinvertebrates for Each Replicate Sample
Collected at Stations 3 and 4 in the Ohio River, Wheeling,
West Virginia, July-August 1984 C-10
C-8 Numbers of Macroinvertebrates for Each Replicate Sample
Collected at Stations 5 and 6 in the Ohio River, Wheeling,
West Virginia, July-August 1984 C-11
C-9 Numbers of Macroinvertebrates for Each Replicate Sample
Collected at Stations 7 and 8 in the Ohio River, Wheeling,
West Virginia, July-August 1984 C-1 3
C-10 Analysis of Variance and Tukey's Studentized Range Test for
Zooplankton, Ohio River C-15
C-11 Analysis of Variance and Confidence Interval-Overlap Results
of Chlorophyll a and Biomass Measurements of Periphyton,
Ohio River C-1 5
C-1 2 Analysis of Variance and Tukey's Studentized Range Test
Results for Oligochaetes and Amphipods, Ohio River C-1 6
C-13 Analysis of Variance and Tukey's Studentized Range Test
Results for Chironomidae Taxa, Ohio River C-16
C-14 Analysis of Variance and Tukey's Studentized Range Test
Results for Trichoptera, Ohio River C-19
C-15 Analysis of Variance and Tukey's Studentized Range Test
Results for the Benthic Macroinvertebrate Taxa, Ohio River C-1 9
VIII
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A ckn o wldgem ents
The assistance of EPA's staff at the Wheeling Office, Region III, with the pre-site
visit, sample collections, substrate sampling and onsite testing is hereby
acknowledged. In addition, the help of the West Virginia Department of Natural
Resources with the pre-site visit and sample collections is gratefully acknowl-
edged. Staff from the State of Ohio Environmental Protection Agency provided
assistance with the selection of the study site.
-------
List of Contributors
LABORATORY TOXICITY TESTS
Scott E. Heinritz1 and Donald I. Mount1
ZOOPLANKTON COMMUNITY
Randall B. Lewis2 and Sharon K. Gross3
PERIPHYTIC COMMUNITY
Randall B. Lewis2 and Ronald J. Bockelman"
BENTHIC MACROINVERTEBRATE COMMUNITY
Randall B. Lewis2 and Alexis E. Steen3
COMPARISON OF LABORATORY TOXICITY DATA AND
RECEIVING WATER BIOLOGICAL IMPACT
Donald I. Mount1
PRINCIPAL INVESTIGATOR: Donald I. Mount1
'Environmental Research Laboratory, U.S. Environmental Protection Agency, 6201 Condgon Blvd., Duluth, MN
55804
:EA Engineering Science and Technology, 612 Anthony Trail, Northbrook, IL 60062.
3EA Engineering, Science, and Technology, Inc.. Hunt Valley/Loveton Center, 15 Loveton Circle, Sparks, MD
21152
'EA Engineering, Science, and Technology, Inc , 221 Oakcreek Drive, Westgate Park, Lincoln, NE 68528
-------
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
NPDES 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 seventh in a series of eight and was conducted on the Ohio
River near Wheeling, West Virginia, which receives discharges from many
industrial facilities, including large steel mills. The study area comprises about
12 km of the Ohio River upstream from Wheeling, West Virginia, in the Pike
Island pool. The Ohio River is a major inland waterway and is navigable
throughout its length. Ambient toxicity tests were conducted on samples from
eight river stations. Biological studies were conducted at these stations and
included plankton, periphyton, and benthic macroinvertebrates.
This site study did not involve effluent testing as a requisite because it was
impractical to do dye dilution studies. Without them, there was no way to use
effluent toxicity data to predict instream impact. Effluent tests were planned
however for use of the State agency. Due to both a problem in sample acquisition
and a mistake in procedure, none were completed.
The impact in the river was not large but all indicators suggest some impact at
Stations 2 and 3. The toxicity to Ceriodaphnia of samples from these two stations
was lowest at these stations although not statistically significant. Fathead
minnow toxicity was lowest at Stations 5 and 6 but the difference compared to
the station with least toxicity was no larger than between duplicates.
The percent of correctly predicted stations ranged from 63 to 100 depending on
the degrees of impairment compared. The Ceriodaphnia data gave exactly the
same profile as the field macroinvertebrate data for species richness. Toxic
impact is most difficult to predict in sites such as this one where the receiving
water is large and the impact is not severe.
XI
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Quality Assurance
Coordination of the various studies was completed by the principal investigator
preceding and during the onsite work. A reconnaissance trip was made to the
site before the study and necessary details regarding transfer of samples,
specific sampling sites, dates of collections, and measurements to be made on
each sample were delineated. The principal investigator was responsible for all
Quality Assurance-related decisions. All instruments were calibrated by the
methods specified by the manufacturers. For sampling and toxicity testing, the
protocols described in the referenced published reports were followed. Where
identical measurements were made in the field and laboratory, both instruments
were cross-calibrated for consistency.
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7. Introduction
The study site was the Ohio River near Wheeling,
West Virginia. One large steel mill with multiple
discharges for a total of approximately 7.3 mVsec
was located within the study area. The Ohio River is
already large even this far upstream and the study
area consisted of a tiny fraction of the river along one
shore. Thinking of the study area as a mixing zone of a
large discharge would give a representative picture.
There are dozens of discharges upstream of the study
area and the water quality entering the study area
contained an unknown amount of effluents from
these discharges. There was no plan to attribute any
ambient toxicity measured to a scjrc^, rather the
objective was to compare the ambient toxicity to
community response in a large river system where
there are many discharges. Previous studies com-
pleted in the study area had revealed reduced
numbers of macroinvertebrates collected in artificial
substrates downstream of a large steel mill complex.
Effluent dilution tests of the steel mill were planned,
but problems with sample acquisition and a random-
ization error required that these test results be
disregarded. Since the intent was to compare ambient
tests to community response, this problem did not
affect the study objectives.
Several of the stations were located in the zone of
effluent mixing as judged by color and temperature.
The discharges and dilution volume were so large
that dye studies were too expensive for the funds
available. The Ohio River is very turbulent and,
without elaborate dye studies, the effluent concen-
trations at various stations cannot even be approxi-
mated. Therefore, the effluent dilution test results
could not have been used to predict impact since
effluent concentrations at the sampling station were
not known. The river flow variation was large when
the substrates were in place, and there was no
information as to how different flows affected the
effluent concentrations at the sampling stations.
Thus, the effluent exposure the substrates exper-
ienced before and after the toxicity test period may
have been the same as, or quite different from the
exosure concentrations during the test period.
Determining the impact of individual discharges to
large rivers using stream surveys is very difficult
unless the impact is dramatic. However, for rivers
such as the Ohio River with many discharges, the
combined effects could be quite large even though
any single discharge would not have measureable
effects on the aquatic community. A method is
needed to assess such "undramatic" individual
discharge effects. If it can be shown that ambient
toxicity texts as used in this study are indicative of
biological response, then there is some better justifi-
cation for using effluent dilution tests to predict
adverse effects even though those adverse effects
from a single discharge cannot be measured by
biological surveys.
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 7. All methods and supporting
data are included in the appendixes for reference.
7-7
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2. Study Design and Site Description
The study area was on the upper Ohio River between
Ohio and West Virginia and included about 1 2 km of
the Ohio River upstream from Wheeling, West
Virginia, in the Pike Island pool (Figure 2-1). The Ohio
River is a major inland waterway and is navigable
throughout. The Ohio River receives effluents from
publically owned treatment works (POTWs), heavy
industry, chemical plants, power generating stations,
and steel mills. Within the study area, there was only
a steel mill with multiple outfalls and a POTW.
Upstream from this part of the river were many
different types of dischargers including power plants,
oil refineries, POTWs, and other steel mill installa-
tions.
Study components included 7-day Ceriodaphnia
dubia toxicity tests and 7-day larval growth tests
using fathead minnows (Pimephates promelas) on
ambient samples from the river stations during 17-23
July. Water samples for the toxicity tests were
collected near the locations of the artificial substrates.
Quantitative assessment of the planktonic, periphytic,
and benthic macroinvertebrate communities was
conducted 5 July to 2 August 1984.
Kilometers
0
Station River Kilometer
3 6
Discharges
River Kilometers
' Steel Mill A
' B
ป C, D. E
POTW
99.4, 99.6
101.2
107.0
106.7
ngs
Creek
8
West Virginia
Figure 2-1. Study area on the Ohio River near Wheeling,
West Virginia. Station locations are indicated.
Stations were also used to collect samples for
zooplankton, periphyton, and benthic macroinverte-
brates. The stations were located upstream, in and
downstream of the effluent plumes which could be
discerned in some areas by visible currents or color.
At each station samples were obtained from two
depths (0.6 and 1.5 m), since the steel mill discharge
was warmer than ambient river temperatures and
vertical mixing might be inhibited. Ambient water
quality measurements in the field were not made. The
stations descriptions are:
Station 1 (RK 97.2)Approximately 1.6 km down-
stream of a POTW. offshore approximately 26 m
from the right bank, water depth 4.5 m. Artificial
substrates were attached to the superstructure of a
wrecked barge. The river bank was gravel and the
river bottom was compacted sediment and rubble.
Station 2 (RK 99.6)Downstream of the first set of
the large steel mill outfalls, offshore approximately
1 2 m from the left bank, water depth 3 m. Artificial
substrates were attached to an icebreaker and
mooring cable. The river bank was concrete and the
bottom was uncompacted organic material.
Station 3 (RK 101.4)Downstream of the second set
of steel mill outfalls, offshore approximately 7 m
from the left bank, water depth 3 m. Artificial
substrates were attached to mooring piers. The
river bank was clay and the bottom was mud.
Sfar/o/?4(RK 104.1)At a marina, approximately 8 m
offshore from the left bank, water depth 2 m. The
artificial substrates were attached to the floating
dock. The river bank and bottom were composed of
mud.
Station 5 (RK 106.7)Approximately 1 km upstream
of a POTW, 14m offshore from the left bank, water
depth 2 m. The artificial substrates were attached
to a fallen tree. The river bank and bottom were
composed of mud.
Station 6 (RK 106.8)Farther downstream of the
POTW, approximately 14 m offshore of the left
bank, water depth 2 m. The artificial substrates
were attached to styrofoam floats. The river bank
was composed of mud, whereas the river bottom
was composed of sand and gravel.
2-1
-------
Station 7 (RK 106.9)Immediately downstream of
the confluence with Harmon Creek which receives
the third set of steel mill discharges. The station
was approximately 27 m offshore of the left bank,
water depth 3 m. Artificial substrates were attached
to styrofoam floats. The river bank was rock fill and
the bottom was mud.
Station 8 (RK 109.4)Downstream of Harmon Creek
by about 2.7 km, offshore approximately 14 m from
the left bank, water depth 4 m. The artificial
substrates were attached to styrofoam floats. The
river bank was stone and the bottom was com-
pacted sediment and rubble.
2-2
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3. Ambient Toxicity Tests
The purpose of the toxicity tests was to measure the
response of Ceriodaphnia dubia and fathead min-
nows (Pimephales promelas) exposed to ambient
Ohio River water. The Ceriodaphnia toxicity tests
measured reproductive potential (number of young
per female) and survival. The fathead minnow tests
measured the weight gain and survival of fathead
minnows. Test results are to be compared with the
macroinvertebrate populations on artificial substrates.
Samples of Ohio River water were collected daily for
seven days from two depths at each of eight stations
located upstream and downstream of a set of large
steel mill discharges on the Ohio River near Wheeling,
West Virginia. Ceriodaphnia and the fathead min-
nows were exposed to each sample for a 24-hour
period and test water was renewed daily with new
sample water. This procedure was used to approxi-
mate the continual exposures which would have
been received had the test organisms been in the
river and to approximate the exposure conditions
where the artificial substrates were suspended.
Descriptions of the sample collections, test methods,
and statistical analyses are provided in Appendix A.
3.1 River Flow Measurements
The Ohio River flow data were used to estimate the
relative effluent dilution and monitor the water flow
over the study area. At stable river flows, a constant
dilution of the effluents at each station would occur.
River flows recorded daily by the National Weather
Service are shown in Table 3-1. The flow data covers
the entire period when the artificial substrates were
in the Ohio River. Mean upstream river flow during
toxicity testing (17-23 July} at East Liverpool (RK
69.2) was approximately 603 mVsec and similarly
downstream at Wheeling (RK 144.8) was approxi-
mately 625 mVsec. The volume flow through the
study area changed over time such that the mean
river flows during the toxicity testing were midway
(603 mVsec) between the extreme flows. The pre-
test mean flows were 227 percent, and the post-test
flows were 52 percent of the flows during the toxicity
test. As a result of these changing river flows, the
exposure of the artificial substrates to effluent
concentrations differed from the exposure of Cerio-
daphnia and fathead minnows. Effluent concentra-
tions in the Ohio River would have been much
reduced in early July during the period of high flow.
Table 3-1. Ohio River Flow (mVsec)
1984
5 Jul
6 Jul
7 Jul
8 Jul
9 Jul
10 Jul
11 Jul
12 Jul
13 Jul
14 Jul
15 Jul
16 Jul
Pre-Test Mean
17 Jul
18 Jul
19 Jul
20 Jul
21 Jul
22 Jul
23 Jul
Ambient Toxicity Testing
Period Mean
24 Jul
25 Jul
26 Jul
27 Jul
28 Jul
29 Jul
30 Jul
31 Jul
1 Aug
2 Aug
Post-Test Mean
Mean (5 Jul - 2 Aug)
East Liverpool
(RK 69.2)
765
898
1,022'"
841"'
1,127
1,045
2,336
2,413
2.166
1,546'"
1,218""
1,014
1,366
844
807
719"'
600
473'"
416'"
365
603
362
374
280
303
323'"
320'"
314
306
297
272
315
819
Wheeling
(RK 146.4)
773
906
1,051'"
886 ">
1,175
1,062
2,271
2,472
2,249
1,626"'
1 ,232'"
1,053
1,396
872
838
733'"
631
491'"
430"'
377
625
371
394
289
306
328'"
331'"
328
314
306
283
325
841
'"Projected flows.
Note: Flows recorded by National Weather Service.
The effluent concentrations to which the substrates
were exposed increased as the flow decreased with
concentrations probably highest after the toxicity test
period, as the flow of the river decreased.
3.2 Chemical and Physical
Test Conditions
Temperature for the Ceriodaphnia tests was main-
tained at 25 ฑ 1ฐC. The fathead minnows were at
3-1
-------
temperatures determined by room temperature which
ranged from 22-28ฐC. Most of this range was caused
by the heat from lights during the daylight period.
Vigorous air mixing assured uniform temperatures
for all chambers at any one time and the water
temperature changes were gradual when the lights
came on and off in the morning and evening. Routine
water chemistry measurements included pH, dis-
solved oxygen (DO), and conductivity for the Cerio-
daphnia and fathead minnow tests (Table 3-2). Initial
values of pH and DO for both test species were 6.6-
74 and 7.9-8.2 ing/liter, respectively. Final values of
pH were slightly higher than the initial values, ranging
7 0-75 for ine fathead minnows and 7.1 -7.7 for the
Ceriodaphnia. Final values of DO were at least 6.6
nig liter for the fathead minnows and at least 7.4
mg liter for the Ceriodaphnia. The conductivities
ranged from 210 to 292 umhos for the 0.6 m samples
and from 263 to 286 umhos for the 1.5-m samples.
3.3 Ambient Toxicity Test Results
At each of eight stations, two water samples were
used for the tests: samples collected at 0.6 m were
identified T and 1.5-m depth samples were identified
B In addition, duplicate tests were conducted using
the 06-m samples from Stations 1, 4, and 8 using the
fathead minnows and are referred to as "A" samples.
Duplicate tests using Ceriodaphnia were conducted
only at Station 1 at both depths.
For statistical comparison, a reference must be used.
Stations T-1 and B-1 were selected for the fatheads
because mean survival was near the highest and
mean weight was the highest at T-1 and the weight of
B-1 was within weighing error of the highest, B-8.
Use of a T sample and a B sample from different
stations did not seem reasonable in view of the small
differences.
Mean survival of fathead minnows varied between 53
and 100 percent for the 0.6-m (T) samples (Table 3-3).
The lowest survival at Station T-7 was significantly
different when compared to Station T-5. Mean
survival of fathead minnows for the 1.5-m(B) samples
ranged from 75 to 95 percent and no significant
differences were found when compared to Station
B-1. The duplicate test results of the 0.6-m (T)
samples were very similar for Stations T-1 and T-4,
with the mean survivals varying by 7 and 5 percent,
respectively (Table 3-3). The duplicate test results for
Station T-8 (comparing T-8 and T-8A) varied by 16
percent.
The mean fathead minnow weights varied only from
0.259 to 0.406 mg (Table 3-4). The ranges for the
0.6- and 1.5-m depths were very similar. The 0.6-m
stations were compared to the highest value T-1; and
four stations (T-1 A, T-4, T-5 and T-7) were significant-
ly lower. However, T4 had a duplicate value that was
not significantly different and the duplicate of T-1A
(T-1) had the highest mean weight. Of the 1.5-m
Table 3-2. Water Chemistry Data for Ambient Toxicity Tests with Fathead Minnows and Ceriodaphnia. Ohio River, Wheeling,
West Virginia, July 1984
Conductivity
(umhosl
268
284
284
265
210
267
266
292
272
-
263
286
285
268
265
264
272
271
Fathead Minnow
and Ceriodaphnia
Initial pH Range
7.0-7.4
-.
7.0-7.2
6.8-7.1
6.8-7 2
7.0
6 772
68-7.2
67-7.4
6.8-7.2
--
6.9-7.1
7.0-7.2
6.7-7.1
6.9-7.1
67-7.0
6.7-7.0
6.8-7.1
6.6-7.1
Fathead Minnow
and Ceriodaphnia Fathead Minnow
lnitialDO Fath^ri Minn, F'nal D0
Mean
|mg/L)
8.2
7.9
7.9
8.1
78
80
7.9
7.9
7.9
--
8.0
7.9
8.0
80
8.0
8.0
8.0
7.9
Range
(mg/LI
78-8.7
_
7.5-8.2
7.8-8.1
7.9-8.5
7.6-8.4
7.5-8.5
7.5-8.3
76-8.3
--
7.8-8.3
7.7-8.3
7.8-82
7.6-8.5
7.8-85
78-8.5
78-8.4
7.0-8.3
Final pH
Range
7.1-7.5
7.0-73
7.1-7.4
7 1-7.5
70-7.4
7.1-7.3
7.0-7.4
7.0-7.4
7.1-7.4
7.1-7.4
7.1-73
7.1-7.4
7.0-7.4
7.0-7.4
7.0-7.4
7.0-7.3
7.0-7.4
7.0-7.4
7.0-7 3
Mean
(mg/LI
6.8
6.7
6.7
6.8
6.7
6.7
6.6
6.8
66
6.6
6.7
6 7
6.7
6.8
6.7
67
6.8
6.6
6 7
Range
(rng/L)
6.1 -7 1
6.3-6.9
62-7.0
6.8-8.1
6 1-7.2
6.2-6.9
6.0-7.0
6.1-7.2
6.1-7 0
6.4-7 0
6.2-7 0
6.2-7.0
6270
6.2-7.1
6.4-7.3
6.2-70
6.3-70
6.1 -7 0
6.4-7 0
Ceriodaphnia
Final pH Range
7.4-7.5
7.1-74
7.2-74
7.3-7.4
7.4-7.5
7.4-7.5
--
74-7 5
7.4-7.5
74-7.5
75-7.6
7.5-7.6
7.2-7.6
7.4-7.7
Ceriodaphnia
Final DO
Mean
(mg/L|
7.4
7.4
7.4
7.5
_
76
7.6
7.7
7 6
-
74
7.6
7 4
76
7.6
7.6
7.6
7.4
Range
(rng/L)
7.2-76
7.2-7.6
70-7.8
70-77
7.4-7.8
7.2-7.9
7.4-79
727 9
72-7.6
73-7.9
7.0-7.8
7.3-7.8
7.5-7.8
7.3-7.8
7.4-7.8
7.0-7.8
Station
'-1
T-1A
T 2
T 3
T-4
T 4A
1 5
T-6
~ -1
' 8
T-8A
B-1
B 2
B-3
B 4
6-5
B-6
B 7
B 8
Note Stations T * A. T-4. A, and T-8 A are duplicates.
T indicates samples were collected near surface at 0 6 m and 8 indicates samples were collected near bottom at 1.5 m
3-2
-------
Table 3-3. Mean Survival of Larval Fathead Minnows for
Ambient Toxicity Tests, Ohio River, Wheeling,
West Virginia, July 1984
Station
T-1
T-1A
T-2
T-3
T-4
T-4A
T-5
T-6
T 7
T-8
T-8A
B-1
B-2
B-3
B 4
B 5
B-6
B-7
B-8
A
90
90
80
100
100
100
100
100
70
80
100
100
90
100
90
90
90
100
80
Replicate
B C
100
90
100
100
90
80
100
100
40
100
70
90
80
90
90
90
80
100
80
90
80
100
100
80
100
100
90
50
100
90
100
80
100
90
90
70
80
80
D
100
90
100
50
80
90
100
80
50
100
60
80
60
90
90
80
100
80
60
Mean
95
88
95
88
88
93
100
93
53""
95
80
93
78
95
90
88
85
90
75
'""Significantly different using two-tailed Dunnett's test |P ! 0.05|.
The T ambient stations were compared against T-1, and B
ambient stationswere compared to B-1 in the statistical analysis
Note Stations T-1 A, T-4 A, and T-8 A are duplicates.
T indicates samples were collected near surface at 0.6 m.
B indicates samples were collected near bottom at 1 5 m.
Table 3-4. Mean Individual Weights (mg) of Larval Fathead
Minnows for Ambient Toxicity Tests, Ohio River,
Wheeling, West Virginia, July 1984
Station
T-1
T-1A
T-2
T-3
T-4
T-4 A
T-5
T-6
T-7
T-8
T-8A
B-1
B-2
B-3
B-4
B-5
B-6
B-7
B-8
Replicate Weights
A B C D
0486
0 176
0.459
0382
0.247
0390
0302
0.297
0290
0.404
0.356
0.383
0409
0381
0.382
0.262
0.244
0327
0.335
0.380
0.254
0.367
0.464
0281
0365
0.294
0299
0345
0.315
0354
0427
0.335
0.421
0.414
0280
0.215
0269
0.349
0.344
0308
0.323
0.381
0.256
0325
0287
0.281
0.240
0305
0389
0.421
0.346
0366
0.342
0.256
0.293
0.290
0.469
0.400
0.303
0.330
0.250
0.341
0.346
0.288
0361
0210
0.302
0.356
0.365
0.300
0.339
0.371
0303
0.344
0.469
0.492
Weighted
Mean
0.402
0.259"
0.365
0.386
0.279'a;
0.356
0.293'ai
0.307
0.270""
0.328
0.365
0.400
0.353
0377
0.377
0.274'"
0.277'"
0.344
0406
SE
0024
0023
0024
0.025
0025
0023
0024
0025
0033
0.024
0024
0.025
0028
0025
0.026
0026
0026
0.026
0028
""Significantly different using two-tailed Dunnett's test IP 0 05).
The T ambient stations were compared against T-1, B ambient
stations were compared to B-1 in the statistical analyses.
Note: Stations T-1 A. T-4A. and T-8A are duplicates.
T indicates samples were collected near surface at 0.6 m
B indicates samples were collected near bottom at 1.5 m.
stations, B-5 and B-6 were different from B-1, but
there were no duplicate values for comparison. Since
half of the significantly different values were dupli-
cates of values that were not different, the statistical
differences found have questionable biological impor-
tance.
The mean survival of Ceriodaphnia ranged from 80to
100 percent at the 0.6-m (T) samples (Table 3-5). For
the 1.5-m (B) samples, Ceriodaphnia survival was
greater than 80 percent, except at Station B-2. No
significant differences in survival at either depth for
any stations were found. Ceriodaphnia reproduction
varied between 19.7 and 28.1 mean number of young
per female for the 0.6-m (T) samples and between
21.7 and 28.8 mean number of young per female for
the 1,5-m (B) samples (Table 3-5). Very similar young
production occurred for the two depths. Using the
highest value of young production at each depth for
comparison, differences in the number of young
produced were not significant.
3.4 Discussion
The Ceriodaphnia ambient toxicity test results did not
show any toxic effects for either survival or young
production. There were some statistically significant
differences between fathead minnow survival and
weights which were confounded by the poor replicate
data. For the 0.6-m sample at Station 7, fathead
minnow survival was low, as was the mean weight
which provides some evidence of toxicity at that
location. However, there is no evidence that toxic
effects, if any, are large.
Table 3-5. Mean Young Production and Percent Survival of
Ceriodaphnia for Ambient Toxicity Tests, Ohio
River, Wheeling, West Virginia, July 1984
Mean Number
of Young
Station per Female
T-1
T-1A
T-2
T-3
T-4
T-5
T-6
T-7
T-8
B-1
B-1A
B-2
B-3
B-4
B-5
B-6
B-7
B-8
28 .1
257
19.7
20.5
27.5
249
243
23.9
24.6
224
25.4
23.1
21 7
266
238
249
28.8
248
95%
Confidence
Intervals
22.8-33.4
206-30.8
14.6-24.8
15.9-25.0
23 .1-31.9
21.9-27.7
19.5-29.5
19.2-28.6
21 0-28.2
18.8-26 1
22.9-279
18.2-28.0
18.4-25.0
22.1-31 1
19.9-27 7
21 5-28.3
236-34.1
18.8-306
Mean
Percent
Survival
100
100
100
80
100
80
90
100
100
80
90
66
100
100
100
100
90
90
Note. Stations T-1 A and B-1 A are duplicates. T indicates samples
were collected nearsurfaceat0.6mandB indicates samples
were collected at 1.5 m. There were no significant differences
between stations or levels (P < 0.05).
3-3
-------
4. Plankton Community Survey
The plankton community was investigated by meas-
uring the occurrence and density of organisms in the
Ohio River. Samples were collected at two depths: 0.6
m and at 1.5 m. The primary emphasis was to collect
zooplankton, but algae were also collected and
enumerated. Measures of the number of species and
individuals are used to determine alterations in
composition and/or density. The sampling and anal-
ytical methods are presented in Appendix B; additional
data are included in Appendix C.
4.1 Community Structure
Rotifers were the dominant taxoncmic group and
accounted for the highest zooplankton concentrations
which occurred at Stations 6 and 8 (Table 4-1).
Brachionus was the most common genus of rotifers
and composed 50 percent or more of the rotifers at
each station. Total densities of rotifers varied from
lows of about 20 organisms/liter at Station 1 to over
100 organisms/liter at Station 4. Crustaceans were
collected at all stations, but in low numbers; densities
varied from 0.6 to 6.3 organisms/liter. Nauplii of
cyclopoid copepods composed the majority of the
crustaceans.
Algae represented a very small portion of the total
plankton densities. Algal densities varied from less
than 1 percent to near 10 percent of the total. With
the use of an 80 ^ net the proportion of algae retained
would be small and so the density would be expected
to be low due to sampling method.
4.2 Evaluation of the Zooplankton
Community
The densities of crustaceans and rotifers were lowest
for Station 1 for both depths (Table 4-1). Crustacean
densities at Station 1 were 0.6 and 2.9 organisms/
liter for the 0.6- and 1.5-m samples, respectively.
Rotifer densities were 20.5 organisms/liter at 0.6 m
and 27.2 organisms/liter at 1.5 m for Station 1.
The results of a two-way Analysis of Variance
(ANOVA) on the total zooplankton densities indicated
significant (P < 0.001) differences between stations
and nonsignificant differences between depths. The
results of a two-way ANOVA on the total rotifer
densities were similar and this is not surprising
considering that rotifers were the overwhelming
component of the zooplankton population. Tukey's
Honestly Significant Difference Test on both zoo-
plankton and rotifer densities indicated that Station 1
was significantly different (P < 0.05) from all other
stations. Crustacean densities revealed significant
differences (P < 0.05) between stations and depths.
Using a two-way ANOVA and Tukey's test (Sokal and
Rohlf, 1981) results indicated that Stations 1 and 3
were significantly different (P < 0.05).
The densities of crustaceans and rotifers were lowest
at Station 1 (Figure 4-1). The abundance of rotifers
increased dramatically between Stations 1 and 2 and
this higher abundance level was consistent down-
stream. The steel mill outfalls are located above
Stations 2, 3, and 7. Travel time from Station 1 to 8 is
about 25 hours (Personal Communication, Wheeling
Office, Region III). Any adverse effect due to the steel
mill discharges is probably not measurable within the
time that the organisms traverse the study area.
In contrast to the variability in the density of zooplank-
ton, taxa were not significantly different either
between stations or between depths.
Crustaceans0.6 m
1 201 .,-,-. Crustaceans1.5 m
_l - Rotifers0.6 m
1 "- Rotifers1.5 m
1 00 \ ,^o- -
\ 80
6
?60i
03
40
20-
/'
/
If
12345678
Sampling Stations
Figure 4-1 . Densities of crustaceans and rotifers collected
in the Ohio River near Wheeling. West Virginia
1984.
4-1
-------
Table 4-1. Densities'" (No.
Taxa
Crustaceans
Cyclopoid copepods
Calanoid copepods
Nauplii
Bosmina sp.
Daphnia sp
Eubosmina sp
Diaphanosoma sp
Total crustaceans
Rotifers
Brachionus budapestinensis
B. calyciflorus
B. caudatus
B angularis
B. urceolaris
B. quadridentatus
B havanaensis
B. bidentata
B variabi/is
Keratel/a sp.
Po/yarthra sp.
Trichocerca sp.
Ketticoltia sp.
Platyias sp.
Filmia sp
Monostyla sp.
Euch/ams sp.
Total rotifers
Algae
Cerat/um sp.
C/ostenum sp.
Total algae
Toial density
Total number of
zooplankton taxa"1'
Taxa
Crustaceans
Cyclopoid copepods
Calanoid copepods
Naupli
Bosmina sp
Daphnia sp
Eubosmina sp.
Diaphanosoma sp
Total crustaceans
Rotifers
Brachionus budapestinensis
B. calyciflorus
B. caudatus
B angularis
B. urceolaris
B. quadridentatus
B. havanaensis
B bidentata
B vanabilis
Keratel/a sp.
Po/yarthra sp
/liter) of Plankton Collected from the Ohio River,
Station
0.6 m
0.3
0.1
02
0.6""
4.6
0.6
6.3
0.1
0.1
8.7
0.1
205ICI
0.1
0.1
212
9
Station
0.6 m
1.8
0.3
1.6
0.5
0.1
43
2.3
15.4
14.8
191
0.4
0.5
0.2
180
06
1
1.5 m
1 1
0.3
1.2
0.1
0.2
2.9
0.3
9.5
3.6
10.2
0.5
0.2
0.1
2.4
0.1
01
0.2
27.2"-"
0.3
0.3
30.4
15
5
1.5 m
2.7
0.3
2.4
0.7
6.1
1 1
153
106
25 1
1 .5
0.5
0.4
0.9
54.8
0.4
Station
0.6 m
1.4
0.2
1.9
06
4.1
2.2
193
16.1
17.2
30
0.2
16.8
1.1
0.3
0.1
0.2
765
4.1
0.4
4.5
85.1
14
Station
0.6 m
0.7
0.2
1.2
0.2
2.3
2.5
290
231
368
2.1
0.4
0.4
0.1
340
0.1
2
1.5 m
1.6
2.0
0.2
3.8
1.2
19.4
162
204
0.9
0.2
0.1
31.2
0.6
0.6
0.2
91.0
7.4
7.4
1022
13
6
1.5 m
1.6
0.2
1.9
0.2
3.9
1.1
18.4
13.1
23.8
2.8
0.9
0.1
0.1
20.0
02
Wheeling, West Virginia.
Station
0.6 m
12
0.7
3.0
0.9
0.4
0.1
63
0.9
35.8
9.8
22.9
2.6
1.0
0.3
0.3
165
0.7
0.3
0.4
0.1
91.6
4.0
4.0
101.9
18
Station
0.6 m
0.8
02
1 3
0.6
2.9
23
185
21.8
293
2.7
04
0.1
21 8
0.8
3
1 5 m
1.6
0.4
2.5
0.9
0.2
5.6
2.7
378
133
21.2
2.9
0.9
04
18.8
0.3
10
99.3
2.9
0.1
3.0
1079
14
7
1 5 m
1.2
0.1
2.0
0.6
3.9
1.7
21.9
166
32.0
1.9
0.5
0.1
28.2
0.2
. July 1984
Station
0.6 m
1.0
0.1
1 7
07
3.5
0.7
196
169
27.2
21
1.2
0.1
0.1
36.1
0.6
0.2
0.3
0.1
105.2
10.2
10.2
1189
16
Station
0.6 rr
0.3
1.7
0.2
2.2
1.8
22.2
32.8
358
1.4
0.4
0.2
0.1
0.1
40.6
0.6
4
1.5 m
1 4
04
1.1
0.9
0.1
3.9
1.4
21.3
12.8
31.6
2.9
0.5
275
0.1
0.3
0.1
986
4.6
0.2
4.8
107.3
15
8
1.5 rn
2.4
03
2 1
0.1
4.9
0.2
195
24.3
24.5
2.7
04
36.7
0.2
4-2
-------
Table 4-1. (continued)
Station 5 Station 6 Station 7 Stafon 8
Taxa
Tnchocerca sp
Kellicott/a sp.
P/atyias sp
Fi/inia sp.
Monosty/a sp
Euchlams sp.
Total rotifers
Algae
Ceratium sp
Closterium sp.
Total algae
Total density
Total number of
zooplankton taxa "'
0.6 m
0.2
0.1
0.4
72.0
3.7
3.7
80.0
16
1.5 m
0.4
0.2
0.1
0.2
11 1.5
8.2
0.1
8.3
125.9
17
0.6 m
0.2
0.1
1288
15.3
15.3
146.4
15
1.5 m
0.1
80.6
2.9
2.9
87.4
14
0.6 m
0.1
0.1
0.1
98.0
12.3
0.1
12.4
113.3
15
1.5 m
0.1
103.2
10.6
10.6
117.7
13
0.6 m
0.5
0.2
136.7
126
12.6
151.5
15
1 5 -n
1.0
0.2
0 1
1098
4.6
46
119.3
5
'"'Density estimates are based on one sample from each location.
'"'ANOVA and Tukey's test indicated Station 1 is significantly different from Station 3 (P = 0.05).
"-'Comparison by ANOVA and Tukey s test indicated Station 1 is significantly different frorr all other stations (P - 0.05|.
""Total number of taxa does not include crustacean nauplii or algae, and there were not significant differences bet ween stations or depths
4-3
-------
5. Periphyton Community Survey
This study investigated the periphytic community by
measuring chlorophyll a and biomass. The relatively
short reproduction time and rapid growth of periphytic
algae results in quick response to changes in water
quality A change in the periphytic community maybe
either a reduction of an important habitat or food
source for other organisms or the enhancement of
nuisance species of algae that neither support higher
trophic levels nor are aesthetically pleas ing. Sampling
and analytical methods are presented in Appendix B.
5.1 Chlorophyll a and Biomass
Measurements
Samples for chlorophyll a and biomass determina-
tions were collected from artificial substrates on 2
August 1984 at a depth of 1.5 m. None of the sample
replicates at Stations 1 and 5 were recovered.
Chlorophyll a replicate values ranged from 1.9 to
151.6 mg/m2 (Table 5-1). The variations within
stations may be due to stream conditions, habitat
availability, or sampling conditions. Mean chlorophyll
a values ranged from 29.1 to 151.6 mg/m2. Three
upstream stations (Stations 2, 3, and 4) had similar
values of 29.1-40.1 mg/m2, whereas the three
downstream stations (Stations 6, 7, and 8) had
higher values of 73.1 -1 51.6 mg/m2. Results of one-
way Analysis of Variance (ANOVA) indicated that
there were significant differences (P = 0.008) in
chlorophyll a between stations when all data were
considered. When Station 8 was omitted, because
there was just one replicate (with the highest value), a
significant difference (P = 0.014) between stations
was still found (Table C-11)
Periphyton biomass varied from 2.4 to 17.4 g/m2
measured as ash-f ree dry weight (AFDW) (Table 5-1).
Similar to the trend with chlorophyll a data, the
biomass at Stations 2 through 4 (3.3-5.8 g/m2) was
generally lower than at Stations 6 through 8 (5.8-
11.1 g/m2).
Results of a one-way ANOVA, using natural log-
transformed data, indicated that the differences in
AFDWs between stations were significant (P = 0.04),
with or without Station 8 data (Table C-11).
Table 5-1. Chlorophyll a and Biomass Measurements of Periphyton Collected from Artificial Substrates in the Ohio River Near
Wheeling, West Virginia, August 1984
Parameter
Sampling Station''
Chlorophyll a frng/m')
Rep 1
Rep 2
Rep 3
Mean
Biomass (m/m?)"'
Rep 1
Rep 2
Rep 3
Mean
Autotrophic Index'17'
Rep 1
Rep 2
Rep 3
Mean
53.8
26.5
40.1
4.1
7.4
5.8
76
276
176
47.2
1.9
44.7
31.2
3.9
2.8
4.1
3.6
82
1.469
92
547
10.5
143
623
29.1
2.4
2.5
5.1
3.3
228
172
82
161
147.2
130.4
89.9
122 5
17.4
84
7.1
109
118
65
79
87
91.6
35.0
92.7
73.1
8.0
3.2
6.2
5.8
87
91
67
82
151 6
151.6
1 1.1
11.1
73
73
la'Dash indicates thai the substrate was missing.
""Measured as ash-free dry weight (AFDW).
cc'Weber 1973.
5-1
-------
Values of an autotrophic index (Al) were calculated
following that of Weber (1 973), and were based on
the ratio of AFDW to chlorophyll a. The Al values
indicatethat heterotrophic(nonalga!)taxa or nonliving
organic matter dominated at Stations 2-4, whereas
autotrophic (photosynthetic) taxa dominated at
Stations 6-8 (Table 5-1).
5.2 Evaluation of the Periphytic
Community
There is a difference in the chlorophyll a content and
biomass for the periphytic community above and
below Station 5. This transition area between Stations
4 and 6 covers a I most 3 km and, unfortunately no data
were available for Station 5. Chlorophyll a values
increased downstream of Station 5. These increases
suggest a source of enrichment between Stations 4
and 6, especially sincethe community downstream of
Station 5 is dominated by photosynthetic taxa.
Potential sources are a POTW located downstream of
Station 5 and Harmon Creek, which receives some of
the steel mill discharges. Station 7 is located down-
stream of the confluence of the Ohio River and
Harmon Creek. However, the two other steel mill
outfalls are located above Stations 2 and 3 where
lower chlorophyll a values were obtained.
5-2
-------
6. Macroinvertebrate Community Survey
This survey investigated the macroinvertebrate
community along the Ohio River using artificial
substrates. Substrate samples were collected at two
depths (0.6 m and 1.5 m) for eight stations. The
benthic community is considered to be a good
indicator of changes in water quality due to restricted
mobility. The degree of community stability can be
ascertained by measuring species composition and
dominance. An alteration in community structure,
species composition, or biomass beyond normal
variations would be regarded as an adverse effect.
Adescription of the sampling and analytical methods
is presented in Appendix B. Additional data are
included in Appendix C.
6.1 Community Composition
The macroinvertebrate community along the study
area on the Ohio River was composed of 56 taxa. The
number of taxa at each station, including the 0.6 m
and 1.5 m substrates, ranged from 13 to 34 (Tables
C-2 through C-5). Two taxonomic groups were
extremely abundant: oligochaetes (unidentified
Naididae) and amphipods (Gammarus sp.) (Table 6-1).
These two macroinvertebrate taxa often composed
over 50 percent of the population. Another seven taxa
which contributed >5 percent of the populations for
at least one station were: chironomids (Cricotopus
cylindraceus group, Dicrotendipes sp., Polypedilum
convictum type, Rheotanytarsus sp., and unidentified
chironomid pupa), hydropsychids (Hydropsyche orris],
and polycentropodids (Cyrnellus fraternus) (Table 6-
1).
The macroinvertebrate community from this area of
the Ohio River is not diverse. Of the nine major taxa of
the community, five are in the Chironomidae family
(midges) and two are in the Trichoptera order
(caddisflies). So, seven of the nine major taxa are
insects, and the remaining taxa are the two most
Table 6-1. Mean Percent Composition of Major Macroinvertebrate Taxa,'" Ohio River, Wheeling, Waซt Virginia
Sampling Stations
Taxa
1
Oligochaeta
Unidentified Naididae 6.0 43.5 38 8 37.8
Amphipoda
Gammarussp. 48.0 40.8 426 18.4
Trichoptera
Hydropsyche orris 0.6 0.0 0.0 1 .8
Cyrnellus fraternus 0.9 0.1 1.7 0.7
Chironomidae
Cricotopus cy/indraceus
group
Dicrotendipes sp
Polypedilum convict urn
type
Rheotanytarsus sp.
Chironom/dae pupae
(unidentified)
Total Chironomidae""
Total 0 6-m Taxa
Total 1.5-m Taxa
1.8
3.3
2.1
17 1
6.9
39.6
23
34
1.5
1.5
3.4
0.0
0.9
14.2
16
14
0.7
9.0
1.2
0.0
0.7
14.4
14
14
1.1
9.1
5.5
0.4
5.7
394
26
24
5 9
25.4
6.7
74
10.0
7.9
4.4
36
6.4
51.9
26
26
17.3
32 2
2.9
32
2.3
4.7
5.5
8.1
5.7
42.0
26
25
685
10.3
1.9
0 1
1 9
1.3
0.9
0.1
26
18 1
20
28
21.9
220
1.8
1.1
5.3
7.0
6 5
2.0
6.7
52.0
25
26
'"Major taxa are those which composed five percent or greater of the total density for at 'east one station The percents are for both
substrate depths.
""Includes all chironomid taxa collected.
Source Tables C-2 through C-9
6-1
-------
abundant: oligochaetes and amphipods. The number
of oligochaete taxa is not known since further
identification was not conducted.
6.2 Station Comparisons
There are noticeable differences in the abundance of
most of the major taxa between stations and depths.
There are also differences in the abundance patterns
between these taxa. Unidentified Naididae densities
varied between depths at Station 7 (located down-
stream of Harmon Creek), and decreased by over an
order of magnitude between Stations 1 and 2 and
Stations 2 and 5 in the 0.6 m samples (Figure 6-1).
Results of an Analysisof Variance(ANOVA) indicated
that differences between depths and stations were
significant (P = 0.001) for the numbers of unidentified
Naididae (Table C-12). Result of Tukey's Honestly
Significant Difference (HSD) test indicated that the
maximum abundance at Station 7 was different than
Stations 1, 3, 4, 5, 6, and 8. Gammarus sp. densities
were greatest at Station 1, then decreased to
minimums at Stations 4 and 7 (Figure 6-2). The
pattern of variation was similar for the two depths.
The ANOVA results indicated that there were signif-
icant differences (P= 0.001) in numbers of Gammarus
between stations, but that the differences between
depths were nonsignificant. The Tukey's HSD test
results indicated that Station 1 was different than
Stations 4, 5, 7, and 8 (P < 0.05).
The densities of each of the five major Chironomidae
were less than 350/m2 at each station, with the
exception of Rheotanytarsus sp. at Station 1 (Figure
6-3). Results of an ANOVA for the abundance of all
chironomid taxa indicated that there were significant
differences(P = 0.0007 (between stations and Tukey's
test results indicated that Stations 2 and 3 were
different (P < 0 05) from Station 8. The patterns of
Oligocnaeia
1 200 Unidentified
Naididae
ซ 0 6 m
1.000' *'* 1 5 m
800-
o
z
I 600'
a
Q
400
200-
3229
Figure 6-1.
2345678
Sampling Stations
Mean density of Oligochaetes (aquatic earth
worms) in the Ohio River.
1,400,
1.200
i.oooj
--. 800-
o !
~ !
I 600 ]
400-
I
200:
Amphipoda
Gammarus sp.
ป-ซ 0.6 m
ป.* 1.5 m
Figure 6-2.
2345678
Sampling Stations
Mean density of Gammarus amphipods in the
Ohio River.
abundance differed for the five major chironomid
taxa. Ail had significant differences between stations
and depths, except that only differences between
stations were found for Dicrotendipes sp. (Table C-
13). for Dicrotendipes sp. Station 2 wasdifferent(P<
0.05) from Stations 3, 4, and 5, and Station 7 was
differentfP <0.05) from Stations 3 and 5. In contrast,
for Polypeditum sp., Station 3 was different (P <0.05)
from Stations 8 and 6. Examination of differences
between stations using Tukey's HSD test indicated
that Station 1 was different (P <0.05} from all other
stations and that Station 6 was different (P < 0.05)
from Stations 2, 3, 4, and 7 for Rheotanytarsus sp.
Further, for Chironomidae pupae Stations 1 and 8
were different (P < 0.05) than Stations 2 and 3 using
Tukey's test. The abundance of Cricotopus cylin-
draceus at Station 5 was significantly different (P :='
0.05) from Stations 4 and 3.
Of the nine major taxa, the two Trichoptera had the
lowest densities. Densities were 80/ m2 or less except
at Stations 5 and 6 (Figure 6-4). In addition, there
were two stations (Stations 2 and 3) where Hydro-
psyche orris was not collected and three stations
(Stations 2. 3, and 7) where Cyrnellus fraternus were
rare. ANOVA results indicatedthat the numbers of H.
orris were significantly different (P < 0.01) between
stations, but were not nonsignificantly different
between depths (Table C-14). The Tukey's HSD test
results indicated that Station 5 was different from
Stations 1, 2, 3, and 4. ANOVA results indicated that
the numbers of C. fraternus were significantly differ-
ent between stations and depth (P< 0.001). Examina-
tion of the differences between stations using Tukey's
test also indicated that Station 5 was significantly
different (P < 0.05) from Stations 2, 4, and 7.
6-2
-------
r 3.000
1.400-1
1,200-
1.000-
600-
400-
200-
Chironomidae
.. Dicrotendipes sp.
OO Rheotanytarsus sp
AA Cricotopus cylindraceus Group
** Polypedilum convictum Type
dD Unidentified Chironornidae Pupae
Total Chironomidae Taxa
-2.500
12 345678
Sampling Stations
Figure 6-3. Mean density of Chironomids (midges) in the Ohio River Densities for each depth are combined.
2001 Trichoptera
-_ Hydropsyche orris
ป-ป Cyrnellus fraternus
160-
120-
f 80
40-
V
2
Sampling Stations
Figure 6-4.
Mean density of Trichopterans (caddiflies) in
the Ohio River. Densities for each depth are
combined.
The macroinvertebrate community was dominated by
the presence of unidentified Naididae and Gammarus
sp, At Station 1, Gammarus sp. contributed 48
percent, while the next most abundant taxon was
Rheotanytarsus sp. At Stations 2 and 3, the unidenti-
fied Naididae and Gammarus sp. each composed
approximately 40 percent of the community. The
contribution to the total abundance from all the
chironomid taxa increased to nearly 50 percent at
Stations 4 through 6. Almost 70 percent of the
community at Station 7 was composed of unidentified
Naididae. In contrast, the unidentified Naididae and
Gammarus sp. were similarly represented at Station
8, each composing about 20 percent of the community
while chironomid taxa again contributed approxi-
mately 50 percent.
6.3 Evaluation of the Macroinvertebrate
Community
Examination of the abundance trends of the major
taxonomic groups indicates that the pattern of
6-3
-------
oligochaete and amphipod density by station appears
to be inversely related. Densities of oligochaetes
were high at stations located immediately down-
stream of the steel mill outfalls (Stations 2, 3, and 7).
In contrast, Gammarussp. had relatively low densities
at these three stations. In addition, at Station 5 where
the trichopterans were relatively more abundant, the
usually very abundant oligochaetes and amphipods
were at a minimum. The five major chironomid taxa
consistently contributed relatively low numbers of
individuals, althoughthe numbers of total chironomid
taxa were much higher and varied greatly between
stations. At the three stations below the steel mill
outfalls, the abundance of the chironomids was
lowest.
The macroinvertebrate community in the upper Ohio
River changes by station; the results of a two-way
ANOVA indicated that there were significant
differences (P = 0.0001) between the number of taxa
per station (Table C-15). However, there were no
significant differences between depths. The total
number of taxa at Station 3 was lowest and is
significantly different (P <0.05) from those at Stations
1, 5, 6, and 8.
6-4
-------
7. Comparison Between Laboratory Toxicity Tests and Instream Biological Response
7.0 Background
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 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 sensitivity of the species in the
community is 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 by
direct toxic effects. Conversely, if the test species is
tolerant of the 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
organisms in the community if the test species is
chosen without knowledge of its sensitivity (as was
the case here).
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 if the reduction is large enough. Therefore, a
lower number of taxa should be a predictable
response of the community. For example, thereshould
be a relationship between the number of young per
female Ceriodaphnia or the growth of fathead min-
nows (or other test species) and the number of
species in the community. Obviously, the test species
must have a sensitivity, such that at ambient concen-
trations 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 fathead minnows
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 may
7-1
-------
be affected at one place near the discharge and a
different group of species may be affected from the
same effluent at another location.
An effluent cannot be viewed as just diluting as it
moves away from the outfall. In fact it is a "series of
new effluents" 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 the report of Mount
et al., 1984; Mount et al., 1985; and in the studies in
press, it is clear that there is no one community
component that is consistently sensitive. 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 other fish to display the
same relative sensitivity to different effluents.
Any test species should have a sensitivity representa-
tive 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 community
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 comprehen-
sive 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. Use of artificial substrates should
reduce habitat effects compared to natural substrates.
They cannot be eliminated. Identification of taxa to
different levels can reduce the sensitivity of species
richness.
Even though species richness has numerous sources
of error as a representative measure of community
health, it remains the best measure for comparison
with lexicological data. Species sensitivity will
respond in the most direct way to toxic response of
the community with the least interference.
7.1 Comparison of Toxicity Test
Results and Field Data
Only the benthic macroinvertebrate data were used
for comparison to the ambient toxicity test data The
number of species/taxa composing the periphyton
community were not determined and so these data
were not available. The zooplankton community,
while sufficient in number of taxa, is not useful
because of the turbulence and the short distance in
the river and resulting rapid time-of-travel from
Station 1 to Station 8. Only if an effluent was
instantaneously lethal to zooplankton, would there
likely be a measurable effect on the population
sampled in this study because the stressed animals
7-2
-------
would remain suspended due to turbulence and
would not die and decompose in the time required to
travel through the study area. A survey offish species
was not conducted. Furthermore, since the study
area was so small, effects on the fish species need to
be dramatic to be detected in such a large river. An
effect on the fish population would most likely have to
be an avoidance response to be measurable. The
statistical analysis of the number of macroinverte-
brate taxa indicated no significant differences be-
tween 0.6 m and 1 .5 m samples, so the data for each
depth were averaged (Table 7-1 ). Survival of fathead
minnows was not significantly different between
stations except Stat ion T-7, but there were significant
differences in weights between stations. However,
these differences are no larger than those between
duplicates in 0.6 m samples for Stations 1 and 4
water. Therefore, the data have been averaged across
depths since the differences are likely due to experi-
mental variation (Table 7-2). None of the stations
were significantly different for Ceriodaphnia young
reproduction or survival and so they, too, were
averaged (Table 7-3). Using the station with the least
toxicityor the most species as zero percent impact, all
Table 7-1. Number of Macroinvertebrate Taxa Collected
from the Ohio River
Mean Number
Total of Taxa
Number Per Station Percent of
Station/Depth of Taxa (+ SD) Reductions""
1 0.6 24 0
29 0 ฑ: 7.1
1.5 34
2 0.6 16 48
15.0 ฑ 1.4
1.5 14
3 0.6 13 53
13.5 ฑ0.7
1.5 14
4 0.6 26 14
25.0 ฑ 1.4
1.5 24
5 0.6 26 2
28.5 ฑ3.5
1.5 31
6 06 26 12
25.5 ฑ0.7
1.5 25
7 0.6 19 21
23.0 ฑ5.7
1.5 27
8 0.6 25 12
25.5 ฑ0.7
1.5 26
""Using Station 1 as the maximum.
Table 7-2.
Station
T-1
T-1A
B-1
T-2
B-2
T-3
B-3
T A
\ -*+
T-4A
B-4
T-5
B-5
T-6
B-6
T-7
B-J
- 1
T-8
T-8A
Bo
-o
Source: Table
Table 7-3.
Station
T-1
T-1A
B-1
B-1A
T-2
B-2
T-3
B-3
T-4
B-4
T-5
B-5
T-6
B-6
T-7
B-7
T-8
B-8
Fathead Minnow Growth
Water.
Mean
Weight (mg) Station Mean
0.402
0.259 0354
0.400
0.365 0.359
0.353
0386 0.382
0377
0*3 "7O
.0 /y
0.356 0.337
0377
0.293 0.284
0274
0.307 0.292
0277
0270 0307
f~\ *l A A
U.J44
0.328 0366
0 365
0.406
3-4.
Ceriodaphnia Reproduction
Water
Mean Number of Station
Young Per Female Mean
28.1
25.7 25.4
224
25.4
19.7 21.4
23 1
20.5 21.1
21.7
275 271
266
249 244
238
24.3 24.6
24.9
23.9 26.4
28.8
24.6 24.7
24.8
in Ambient Station
Percent Increase
of Toxicity
7
6
0
12
26
24
20
4
in Ambient Station
Percent Increase
of Toxicity
6
21.
22
0
10
9
3
9
Source: Tables C-2, C-3, C-4, and C-5
Source: Table 3-5.
7-3
-------
other stations are calculated as a percent of that
value. Because there were many potential sources of
toxicity upstream of Station 1, that station could not
be considered free from toxicity nor could any other.
Therefore, the station with the least toxicity or the
most number of taxa, was considered least impacted
and was used as zero impact for comparative
purposes.
The percent impact at all other stations was then
calculated from that value and each measurement
(fathead minnow toxicity, daphnid toxicity and re-
duced species richness) used a different reference
station as zero percent impact. Tables 7-1, 7-2, and
7-3 show these values. Table 7-4 was then con-
structed as follows. For each station, if the highest
toxicity percentage and species richness percentage
were each below 20 percent or each was 20 percent
or more, a correct prediction was scored. This number
of correct predictions was entered into the upper left
column of Table 7-4 as a percent value. Similar
calculations were done for each column of the matrix
substituting the appropriate percent values for each.
The 20 percent incremental categories are arbitrarily
selected, The percent correctly predicted stations is
75 percent using the 20-100 petcent for the toxicity
data and the 20-100 field data. It was 63 percent for
the 20-100 percent toxicity data and the 40-100
percent field data. One hundred percent are correctly
predicted using 40-100, 60-100, and 80-100 percent
for the toxicity data and for the field data. The
prediction of these higher impact levels are predic-
tions of no effect because the reductions in both filed
and toxicity data were ot severe enough to causethat
much impact. These data are not sufficient to judge
what percent is the best predictor. After all eight
study site reports are completed, an overall assess-
ment can be made to ascertain which reduction level
is the best predictor of instream biological response.
Figure 7-1 shows the profiles of toxicity, based on
daphnid data, and the percent change in macro-
invertebrate taxa at the eight stations. The profiles
are very similar. If the increased toxicity at Stations 5
and 6 evidenced by the fathead minnow (Table 7-2) is
real and not experimental variation, whether it would
E 80
j Ceriodaphnia
Taxa
60
40
20 r
a 12345678
Stream Station
Figure 7-1. Percenttoxicity and percent reduction in macro-
invertebrate taxa for eight ambient stations.
be evidenced by other groups of organisms not
enumerated in this study cannot be judged. The
profiles of fathead minnow data and macroinverte-
brates are not similar.
The much higher river flows (about 2 times) during
the toxicity testing period probably substantially
lessened the effluent exposure in the toxicity tests
compared to the effluent exposure the macroinverte-
brate substrates received during the last 10 days they
were in the river.
There is no evidence of gross toxicity in either the field
or the laboratory data. The Ceriodaphnia data show
the most toxicity at Stations 2 and 3 and the
macroinvertebrates show the greatest reductions
there as well. The fathead minnow data show the
most toxicity at Stations 5 and 6. Considering the
limited field data for comparison and the large river
size, the ambient toxicity data are reasonable esti-
mates of instream biological response, where the
toxic effects, if present, are not dramatic.
Table 7-4. Percent of Stations Correctly Predicted Using
Four Categories of Percent Impact
Toxicity
Data
20-100
40-100
60-100
80-100
20-100
75
75
75
75
Field
40-100
63
75
75
75
Data
60-100
75
100
100
100
80-100
75
100
100
100
Source Tables 7-1, 7-2, and 7-3.
7-4
-------
References
Hamilton, M. A. 1984. Statistical Analysis of the
Seven-Day Ceriodaphnia reticulata Reproductivity
Toxicity Test. EPA Contract J3905NASX-1. 16
January. 48 pp.
Mount, D. I. and T. J. Norberg. 1 984. A seven-day life
cycle cladoceran toxicity test. Environ. Toxicol.
Chem. 3(3):425-434.
Mount, D. I. and T. J. Norberg-King, ed. 1985. Validity
of Effluent and Ambient Toxicity Tests for Predicting
Biological Impact, Scippo Creek, Circleville, Ohio.
EPA/600/3-85/044.
Mount, D. I., A. E. Steen, and T. J. Norberg-King, ed.
1985. Validity of Effluent and Ambient Toxicity
Tests for Predicting Biological Impact in Five Mile
Creek, Birmingham, Alabama. EPA/600/8-85/
015.
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/600/3-84/080. August. Various pag-
ings.
Norberg, T. J. and D. I. Mount. 1985. A new fathead
minnow (Pimephalespromelas)subchror\\c toxicity
test. Environ. Toxicol. Chem. 4(5):711-718.
Rogers, J. 1 984. University of Wisconsin at Superior,
Wisconsin, and EPA Environmental Research
Laboratory at Duluth, Minnesota. July. Personal
communication.
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.
Stemberger, R. S. 1979. A Guide to Rotifers of the
Laurentian Great Lakes. EPA/600/4-79/021. 186
PP-
Weber, C. I. 1973. Recent developments in the
measurements of the response of plankton and
periphyton to changes in their environment. In:
Bioassay Techniques and Environmental Chemistry
(G. E. Glass, ed.), pp. 119-138. Ann Arbor Science
Publishing, Ann Arbor, Michigan.
R-1
-------
Appendix A
Toxicity Test and Analytical Methods
Each of eight ambient stations along the Ohio River
was sampled at depths of 0.6 m and 1.5 m. All
samples were collected as daily grab samples using
an electric pump and collected in 1-gal collapsible
polyethylene containers. Samples were collected
daily between 0900 hours and 1500 hours. On 18
July samples could not be collected due to mechanical
problems on the boat.
Samples were filtered through a plankton net to
remove zooplankton. Temperature and dissolved
oxygen (DO) concentrations of the ambient samples
were between 24-26ฐC and 7.9-8.2 mg/liter, respec-
tively. The testing was conducted by the EPA
Wheeling Office, Region III, West Virginia.
A.1 Ceriodaphnia Test Methods
Adult Ceriodaphnia dubia from ERL-Duluth which
were 10 days old were used as brood stock. They were
transported by air to Wheeling and immediately
transferred to fresh Ohio River water. These animals
had been cultured in Ohio River water at ERL-D for
seven days prior to test initiation.
The test met hod generally fol lowed that of Mount and
Norberg (1984) with the exceptoin that 1-oz plastic
portion cups were used instead of glass beakers. The
cups were discarded after use.
Ten replicates were run from each ambient sample
and each cup contained 1 5 ml of sample. Less than
six-hour-old Ceriodaphnia were placed in each
replicate cup; except for five replicates from Stations
T-1 through T-6 at 0.6-m and Stations B-1A and B-4
through B-8 at 1.5 m, where animals less than 24
hours old were used to initiate the tests. Temperature
throughout the test was maintained at 25 ฑ 1 ฐC in
thermostatically controlled incubators. Initial DO, pH,
and conductivity measurements were taken from the
2-liter sample for the fathead minnow test and were
used as initial values for both test organisms.
Test solutions were renewed daily and young, if
present, were counted and discarded. Final DO and
pH were measured in one of ten cups from each
ambient station after each renewal. Samples were
not renewed on 1 8 July. However, survival observa-
tions were recorded for this date.
A food formulation was used which consisted of three
parts: (1) 5 g/liter of dry yeast; (2) 5 g/liter of
CerophylS* stirred overnight and filtered through a
plankton net; and (3) 5 g/liter of trout chow, aerated
vigorously for seven days, settled, and decanted. The
yeast suspension and the supernatant from the
Cerophylฎ and trout chow were mixed in equal parts
every seven days. The mixture was kept refrigerated
as were the Cerophylฎ and yeast components, but the
trout chow supernatant was kept 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 of Ceriodaphnia
rather than 0.05 ml as was recommended for yeast
diet (Mount and Norberg 1 984). The suspended solids
concentration in this food is ~1,800 mg/liter.
Groups of five replicatees from each station and
depth were randomized daily on test boards, but
maintained the same shelf position in the incubators
throughout the test.
A.2 Fathead Minnow Tests
The methods for the fathead minnow tests followed
those described by Norberg and Mount (1 985). Larval
fathead minnows were less than 24 hours old and
were air shipped from the USEPA Newtown Fish
Toxicology Station. The fish were assigned one to four
at a time to replicate compartments until each had 10
fish (or 40 fish per station).
Newly-hatched brine shrimp were fed three times
daily. The uneaten brine shrimp were removed daily
during the renewal process by siphoning the tanks to
a depth of approximately one centimeter, after which
two liters of new test solution were added. To aid in
the renewal, a rubber foot made from a Tygon Y-tube
and attached to the siphon was used during the
renewal.
Before the test solutions were renewed, final DO and
pH measurements were recorded. Room temperature
was maintained between 22-28ฐC. There was a 16-
hour light, 8-hour dim photoperiod throughout the
testing period. Chamber locations were randomized
daily.
"Cerophyl1" was obtained from Agri-Tech, Kansas City, Missouri As of
January 1985, Cerophyl" was no longer being produced by that manu-
facturer Use of trade names does not constitute endorsement.
A-1
-------
On 1 8 July no river water was collected. However,
survival observations were recorded and the test
solutions were siphoned down to approximately one
liter and excess brine shrimp were removed. This was
done to improve the surface-to-volume ratio and
prevent possible BOD stress effects on the fish.
After seven days, the fish were preserved in 4 percent
formalin. Upon returning to Duluth, they were rinsed
with distilled water, oven-dried for 18 hours in pre-
weighed aluminum weighing boats, and weighed on
a five-place analytical balance.
A.3 Quantitative Analyses
A.3.1 Ceriodaphnia
The statistical analyses were performed using the
procedure of Hamilton (1984) as modified by Rogers
(personal communication). In this procedure the
young production data were analyzed to obtain the
mean number of young per female per treatment.
Daily means were calculated and these means were
summed to derive the 7-day mean young value. By
this method, any young produced from females that
die during the test are included in the mean daily
estimate (all data method). Using this procedure,
mortalities of the original females affect the estimate
minimally, but the mortality of the adult is used along
with the young production to determine overall
toxicity. Confidence intervals are calculated for the
mean reproductivity using a standard error estimate
calculated by the bootstrap procedure. The bootstrap
procedure subsamples the original data set (1,000
times) by means of a computer to obtain a robust
estimate of standard error.
Tukey's Honestly Significant Difference test (Sokal
and Rohlf, 1981) is used to determine significant
differences in survival and young production between
stations.
A.3.2 Fathead Minnows
The mean weights are statistically analyzed with the
assumption that the four test chamber compartments
behave as replicates. The method of analysis assumes
that the variability in the mean treatment 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 treatments. The t-statistic is
then compared to the critical t-statistic for the
standard two-tailed Dunnett's test (Steele and Torrie
1960) The survival data are arcsine-transformed
prior to the regression analyses to stabilize variances
for percent data.
A-2
-------
Appendix B.
Biological Sampling and Analytical Methods
B.1 Plankton Survey
Plankton were collected from eight stations on the
Ohio River near Wheeling, West Virginia, on 23 July
1984. Samples were collected at 0.6- and 1.5-m
depths by pumping 10 liters of water through an80-m
mesh net. No sampling replication was conducted.
Samples were preserved in 10 percent formalin. 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.
The crustaceans, rotifers, and total zooplankton were
analyzed by Analysis of Variance (ANOVA) on the
untransformed and natural log-transformed data. A
two-way ANOVA was performed on the densities of
these three groups to determine if there are differ-
ences between stations and depths. In addition, a
two-way ANOVA was performed on the number of
taxa per station. Tukey's Honestly Significant Differ-
ence tests were conducted to determine which
stations were different, when a significant difference
was detected using the ANOVAs.
B.2 Periphyton Survey
The periphytic community was sampled quantitatively
using clear acetate strips suspended in the Ohio River
at the same locations as the 1.5-m artificial substrates
for the benthic macroinvertebrates. Triplicate strips
were placed in the river at eight stations on 5 July
1984 and retrieved on 2 August 1984 for a 28-day
colonization period.
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 105ฐC to a constant
weight and ashed at 500ฐC. Distilled water then was
added to replace the water of hydration lost from clay
and other minerals. Samples were redried at 105ฐC
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-
fuged and analyzed spectrophotometrically. A
chlorophyll a standard (Sigma Chemicals) extracted
in a 90 percent acetone solution was used for instru-
ment calibration. Chlorophyll a standing crop was
expressed as mg/mj. The biomass and chlorophyll a
data were used to calculate the Autotrophic Index
(Weber 1 973), which indicatesthe relative proportion
of heterotrophic and autotrophic (photosynthetic)
components in the periphyton.
The chlorophyll a and AFDW data were statistically
examined by one-way ANOVA using SAS and
MINITAB to detect differences between sampling
locations. The ANOVAs were performed on all data
and again with Station 8 omitted. (Station 8 had the
highest value and only one of the three replicate
substrates was recovered.)
B.3 Benthic Macroinvertebrate Survey
Aquatic macroinvertebrates were sampled from the
Ohio River during July and August 1984 utilizing
Hester-Dendy artificial substrates. The Ohio River
was sampled at eight locations from RK 100 to RK
113 near Wheeling, West Virginia. Samplers were
placed in the river on 5 July 1 984 and retrieved on 2
August 1984, resulting in a 28-day colonization
period. Three replicate Hester-Dendy samplers were
suspended from permanent structures along the
shoreline at 0.6- and 1.5-m depths at each Jocation.
The samplers at the 1.5-m depth were round-plate
substrates as described by Weber (1 973) which have
an effective surface area of 0.13 m2. The samplers at
the 0.6-m depth were square-plate substrates (indi-
vidual plate = 7.5 x 7.5 cm) constructed by the
Wheeling, West Virginia office of the USEPA. The
square-plate samplers were constructed to conform
with the round-plate samplers; however, they had an
effective surface area of 0.1 6 m2. The samplers were
preserved upon retrieval with 10 percent formalin
with rose bengal stain added to aid in sorting.
Macroinvertebrates and debris were scraped and
brushed free of the artificial substrate upon receipt in
the laboratory. The residue and organisms collected
on each sampler were sieved in the laboratory on a
8-1
-------
U.S. Standard No. 30 mesh sieve and preserved in 10
percent formalin. All samples were analyzed utilizing
procedures outlined in EA's Macroinvertebrate
Quality Control and Procedures Manual. Prior to
analysis, each sample was rinsed on a U.S. No. 60
mesh sieve to remove preservative.
The sample material was then sorted, a small portion
at a time, under a dissection microscope at 10X
magnification. All organisms (except chironomids)
were identified under 10X magnification. The chiron-
omids were mounted on glass si ides in a nonresinous
mounting media for examination under a compound
binocular microscope at 40-1,OOOX magnification.
Oligochaeta (segmented worms) were not identified
beyond the familial level. All other organisms were
identified to the lowest taxonomic level practicable
(usually genus or species) using state-of-the-art
taxonomic keys. Abundance was standardized to
number per m2 for density comparisons.
The macroinvertebrate data were analyzed using
two-way ANOVAs on the numbers of organisms for
selected taxa: unidentified Naididae, Gammarus sp.,
Hydropsyche orris, Cyrnellus fraternus. Cricotopus
cylindraceus. Dicrotendipes sp., Polypediium convic-
tum type, unidentified Chironomidae pupa, and total
Chironomidae. The ANOVAs were performed to
detect any differences between stations or depths.
Tukey's Honestly Significant Difference test was
performed when a significant difference was detected
using the ANOVAs to determine which stations were
different. In addition, a one-way ANOVA and Tukey's
test were performed on the total number of taxa per
station.
B-2
-------
Appendix C
Additional Biological Data
Table C-1. Numbers of Plankton Collected from the Ohio River Near Wheeling, West Virginia, August 1984
Station! Station 2'" Stations Station 4
Taxa
Crustaceans
Cyclopoid copepods
Calanoid copepods
Nauplii
Bosmina sp.
Daphnia sp
Eubosmma sp.
Diaphanosoma sp.
Total crustaceans
Rotifers
Brachionus budapestinensis
B calyciflorus
8. caudatus
B. angularis
8. urceolans
8. quadridentatus
B. havanaensis
B. bidentata
B. variabilis
Keratella sp.
Polyarthra sp.
Trichocerca sp
Kellicottia sp.
P/atyias sp.
Fi/inia sp.
Monostyla sp.
Euch/anis sp.
Total Rotifers
Algae
Cerat/um sp
C/ostenum sp
Total Algae
Total Zooplankton
Taxa
Crustaceans
Cyclopoid copepods
Calanoid copepods
Nauplii
J?tfj-,^?<-i^a sp
Daphnia sp
Eubosmina sp.
Diaphanosoma sp
Total Crustaceans
0.6 m
3
1
2
6
46
6
63
1
1
87
1
205
1
1
212
Station
0.6 m
18
3
16
5
1
43
1.5 m
11
3
12
1
2
29
3
95
36
102
5
2
1
24
1
1
2
272
3
3
304
5
1.5 m
27
3
24
7
61
0.6 m
14
2
19
6
41
22
193
161
172
30
2
168
11
3
1
2
765
4
41
45
851
Station
0.6 m
7
2
12
2
23
C-1
1.5 m
16
20
2
38
12
194
162
204
9
2
1
312
6
6
2
910
74
74
1,022
6
1.5m
16
2
19
2
39
0.6 m
12
7
30
9
4
1
63
9
358
98
229
26
10
3
3
165
7
3
4
1
916
40
40
1,019
Station
0.6 m
a
2
13
6
29
1.5 m
16
4
25
9
2
56
27
378
133
212
29
9
4
188
3
10
993
29
1
30
1,079
7
1.5 m
12
1
20
6
39
0.6 m 1
10
1
17
7
35
7
196
169
272
21
12
1
1
361
6
2
3
1
1,052
102
102
1,189 1
Station 8
0.6m 1.
3
77
2
22
.5 m
14
4
11
9
1
39
14
213
128
316
29
5
275
1
3
1
1
986
46
2
48
,073
5m
24
3
21
1
49
-------
Table C-1 (continued)
Taxa
Rotifers
Brach/onus budapest/nensis
B calyciHorus
B caudatus
B dngulans
B urceolans
B quadndentatus
8 havanaensis
B bidentaia
B vanabilis
Keratella sp.
Po/yanhra sp
Tnchocerca sp
Kellicotna sp.
Plalyms sp.
Filinta sp
Monostyla sp.
Euch/anis sp
Total Rotifers
Algae
Ceratium sp.
C/ostenum sp
Total Algae
Total Zooplankton
Station 5 Station 6
06m 15m 06m 15m
23 11 25 11
154 153 290 184
148 106 231 131
191 251 368 238
4 15 21 28
5549
2441
1 1
9
180 548 340 200
6412
242
1 2
1 1 1
4
2
720 1,115 1,288 806
37 82 153 29
1
37 83 153 29
800 1,259 1.464 874
'"Density estimates are based on one sample from each location.
Table C-2. Density (No./m3) and Percent Occurrence of Macroinvertebrates
Wheeling, West Virginia, July-August 1984
Station 1
Taxa
Coelenterata
Hydra sp
Platyhelmmtnes
Planamdae
Dugesia sp.
Annelida
Oligochaeta
Naididae
Unid Na:didae
Crustacea
Amphipoda
Gar-irnandae
Gammarus sp
Acari
Hydracanna
0.6m 1.5m
Mean Mean
No. m! Percent No 'm: Percent
2.1 -:0.1
667 27 487 2.1
1500 6.1 1384 5.9
8396 341 1,481.6 627
2.1 -:0.1 26 01
Station 7 Station 8
0.6 rr, 1,5 m 06m 1 5 m
23 17 18 2
185 219 222 195
218 166 328 243
293 320 358 245
27 19 14 27
4544
1 1 2
1
1
218 282 406 367
8262
0 010
2 2
1
1 1 1
980 1,032 1,367 1,098
123 106 126 46
1
124 106 126 46
1,133 1,177 1,515 1,193
Collected at Stations 1 and 2 in the Ohio River,
Station 2"
0.6 m 1.5 m
Mean Mean
No /m2 Percent No.. m; Percent
21.9 0.9 115 0.7
1,153.1 495 5383 345
765.6 328 8228 527
C-2
-------
Table C-2. (continued)
Taxa
Insects
Ephemeroptera
Heptageniidae
Stenonema sp.
,?. integrum
S terminatum
Caenidae
Caenis sp.
Odonata
Libellulidae
Perithemis sp.
Tnchopterci
Hydropsvchidae
Hydropsyche orris
H simulans
Polycentropodidae
Cyrne/lus fraternus
Neureclipsis sp.
Diptera
Empididae
Unid. Empididae
Chironomidae
Chironomus sp.
Cricotopus bic/nctus group
C. cylindraceus group
C. intersectus group
C. tremuius group
Dicrotendipes sp.
Harnischia sp
M/cropsectra sp.
M. curvicorn/s
Nanocladius sp.
Parametriocnemus sp.
Paratanyrarsus sp
Potypedi/um convictum type
P. fa/lax group
P. scalaenum type
Pseudocfi/ronomus sp
Rheotanytarsus Sp.
Stenochironomus sp.
Tanytarsus sp.
Thienemannimyia series
Unid. Chironomidae pupa
Mollusca
Gastropoda
Ancylidae
Ferrissia sp.
Physidae
Physa sp.
Pelecypoda
Corbiculidae
Corbicula f/uminea
Total Benthos
'oial Taxa""
Station 1
06
Mean
No.'rrv
42
4.2
2.1
16.7
2.1
6.3
6.3
33.3
79.2
4.2
62.5
95.8
41.7
75.0
4.2
833
42
66.7
8.3
202.1
2,462.9
24
m
Percent
0.2
0.2
<:o i
0.7
<0.1
0.3
0.3
1.4
3.2
0.2
2.5
3.9
1.7
3.0
0.2
3.4
0.2
27.1
0.3
82
100
1
Mear
No. m '
26
5.1
2.6
--
2.6
103
2.6
38.5
5.1
2.6
2.6
7.7
_..
2.6
61.5
5.1
17.9
2.6
30.8
12.8
103
17.9
2.6
17.9
5.1
158.9
33.3
5.1
17.9
133.3
2.6
5.1
66.6
2,363.5
34
Station 2la'
.5 m 0.6 m
Mean
Percent No rn-' Percent
0.1
0.2
0.1
0.1
0.4
0.1
1.6
0.2
6.3 0.3
0.1
0.1 6.3 0.3
0.3 53.1 2.3
15.6 07
0.1 18.8 0.8
2.6 50.0 2.1
0.2
0.8 3.1 0.1
0.1
1.3 15.6 0.7
0.5
0.4
0.8 118.8 5.1
0.1 3.1 0.1
0.8 6.3 0.3
0.2
6.7
1.4
0.2
0.8 68.8 3.0
5.6 21.9 0.9
0.1
021
2.8 3.1 0.1
100 2,331.4 100
16
1 5 m
Mean
No m1 Percent
_.
3.8 02
_
-
__
3.8 02
3.8 0.2
3.8 0.2
77 0.5
.._
_.
.__
30.8 2.0
15.4 1.0
23.1 1.5
__
46.1 3.0
.._
346 2.2
11.5 0.7
_.
__
3.8 0.2
1,560.8 100
14
'"One replicate substrate was not recovered.
""There were highly significant differences between stations (P = 0.0001) The number of taxa at Station 3 was different than at Stations 1,
6. and 8 (P = 0.05|.
NOTE: Total Taxa = distinct taxa; does not include pupa of included taxa.
-------
Table C-3. Density (No./ma) and Percent Occurrence of Macroinvertebrates Collected at Stations 3 and 4 in the Ohio River,
Wheeling, West Virginia, July-August 1984
Stations
Station 4
Taxa
Platyhelminthes
Planariidae
Dugesia sp.
0.6 m 1.5 m 06m
Mean Mean Mean
No . m! Percent No. 'm? Percent No /m2 Percent
2.1 0.2 7.7 0.4
1.5
Mean
No./m' Percent
Annelida
Oligochaeta
Naididae
Unid. Naididae
Crustacea
Amphipoda
Garnmaridae
Gammarus sp
Oecapoda
Astacidae
Immature Astacidae
Acan
Hydracarina
Insecta
Ephemeroptera
Baetidae
Baetis sp
Heptageniidae
Stenacron interpunctatum
Stenonema inlegrum
Immature Heptageniidae
Caenidae
Tricorythodes sp
Tnchoptera
Hydropsychidae
Hydropsyche orris
Poiycentropodidae
Cyrnellus fraternus
Neureclipsis sp
Diptera
Empididae
Unid. Empididae
Chironomidae
Ablabesmyia sp
Cricotopus bicinctus group
C cy/indraceus group
C intersectus group
C tremu/Lts group
Dicrotendipes sp.
Endochironomus sp.
Glyptotendipes sp
Micropsectra sp.
Nanoctadius sp.
Orthoclad/us sp
Parachironomus sp
Parametnocnemus sp
Paratanytarsus sp
Phaenopsectra sp.
Polypedilum convictum type
P. fa/tax, group
P. sca/aenum type
Pseudochironomus sp
Rheotanytarsus sp
2979 226 8895 51.0
7354
559
569 1 32 6
0.1
2.6
12.8
0.7
2.6
5.1
0.1
0.3
29.2
2.2
23.1
1.3
2.1
22.9
2.1
12.5
1375
2.1
354
83
0.2
1.7
02
0.9
104
0.2
2.7
0.6
1384
7.9
2.6
26
17.9
0.1
0 1
1.0
618.8
185.4
6.3
2.1
21
18.8
6.3
2.1
21
188
37.5
12 5
122.9
8.3
333
21
4.2
2,1
10.4
2.1
1104
146
2.1
104
45.3
13.6
0.5
02
02
1.4
0.5
0.2
0.2
1.4
2.7
0.9
9.0
0.6
2.4
0.2
0.3
0.2
0.8
0.2
8.1
11
0.2
0.8
256.3
241.0
5.1
2.6
128
23.1
154
2.6
5 1
2.6
7.7
5.1
872
2.6
2 6
7.7
12 8
2.6
179
5.1
97.4
27.0
25.4
0.5
0.3
1.3
2.4
1 6
0.3
0.5
0.3
08
0.5
9 2
0.3
0.3
0.8
1 3
0.3
1.9
0.5
10.3
C-4
-------
Table C-3. (continued)
Station 3
Station 4
Taxa
Stenochironomus sp
Tanytarsus sp
Thienerriannimyia series
Unid. Chironomidae pupa
Mollusca
Pelecypoda
Corbiculidae
Corbicula flummea
Total Benthos
Total Taxa'1"
0.6 m
Mean
No m' Percent
-
_.-
16.7 1.3
125 09
1,316.7 100
13
1.5 m
Mean
No rri'' Percent
-
--
17.9 1.0
10.3 06
41 0 2.4
1,743.2 100
14
06 -n
Mean
No m- Percent
._
83 0.6
37.5 2.7
85.4 6.2
1,366.9 100
26
1
Mean
No m-
30.8
2.6
538
46.1
--
9486
24
5
Percent
3.2
0.3
5.7
4.9
--
100
'"There were highly sign.-fream differences between stations (P- 0.0001}. The number of taxa at Station 3 was different than at Stations 1,
6, and 8(P = 0.05).
Note Total Taxa distinct taxa. does not include pupa of included taxa
Table C-4. Density (No./m2) and Percent Occurrence of Macroinvertebrates Collected at Stations 5 and 6 in the Ohio River,
Wheeling, West Virginia, July-August 1984
Station 5
Station 6
Taxa
PlatyhelTiinthes
Planariidae
Dugesia sp.
0.6 m
Mean
No./m2 Percent
2.1 0.2
1.5 m
Mean
No./m2 Percent
7.7 0.5
06 m
Mean
No./m3 Percent
2.1 0.1
1.5
Mean
No./m' Percent
154 0.7
Annelida
Oligochaeta
Naididae
Unid. Naididae 112.5 8.8 53.8 35
Crustacea
Amphiboda
Gammandae
Gammarus sp. 254.2 19.9 461.4 29.9
Acari
hydracarina -- 15.4 1.0
Insecta
Ephemeroptera
Heptageniidae
Stenacron inierpunctatum 2.1 0.2
Stenonema integrum 2.1 0.2 5.1 0.3
Oaonata
Libellulidae
Perithem/s sp 2.6 0.2
Trichoptera
Hydropsychidae
Cheumatopsyche sp 10.3 0.7
Hydropsyche orris 83.3 6.5 1051 6.8
H. Orris pupa --
H. sitnulans -- 51 0.3
Potamyia (lava 6.3 0.5 -- -
Symphitopsyche morosa -- --
560.4 281 1692 76
5063 25.4 8536 38.2
6.3 0.3
2.1
4.2
33.3
42
2.1
2.1
0.1
0.2
1.7
0.2
0.1
01
26 01
2.6 0.1
897 4.0
128 0.6
C-5
-------
Table C-4. (continued)
Station 5
Taxa
Polycentropodidae
Cyrnellus fraternus
Neurecl/psis sp.
Leptoceridae
Oecetis sp
06
Mean
No m;
31.3
42
.
m
Percent
2.5
0.3
1 5
Mean
No m-
176.9
5.1
2.6
m
Percent
11.5
0.3
0.2
0
Mean
No m-'
63
6.3
--
Station 6
6 rr
Percent
0.3
0.3
1 5
Mean
No rrv
128.2
28.2
--
Percent
5.7
1.3
--
Diptera
Empididae
Unid. Empididae -- 5.1 0.3 5.1 0.2
Chironomidae
Ab/abesmyia sp. 5.1 0.3
Cricotopus bicinctus group 6.3 0.5 70.8 3.6 28.2 1.3
C cylmdraceus group 212.5 16.7 69.2 4.5 72.9 37 256 1.1
C. intersectus group 56.3 4.4 17.9 1.2 8.3 0.4
C. tremulus group 54.2 4.2 17.9 1.2 563 2.8 25.6 1.1
Dicrotendipes sp. 854 6.7 135.9 88 97.9 4.9 1025 46
Glyptotendipes sp 5.1 0.3
Micropsectra sp 8.3 0.7 -- 2.1 0.1 205 0.9
M. curvicornis 2.1 0.2 -- -- -- 2.6 0.1
Microtendipes sp. 2.1 0.2 33.3 22
Hanocladius sp. 771 60 41.0 2.7 375 1.9 897 40
Parametriocnemus sp 2.6 0.2 ~ 20 5 0.9
Paratanytarsus sp. 6.3 0.5 2.6 0.2
Phaenopsectra sp. 42 0.3
Polypedilum convictum type 521 4.1 718 4.7 154.2 7.7 76.9 34
P. fa/lax group -- 2.1 0.1
P. scalaenum type 125 VO 53.8 35 20.8 1.0 538 24
Pseudochironomus sp 2.1 0.2 2.6 0.2
Rheotanytarsussp. 646 5.1 35.9 2.3 122.9 6.2 220.4 9.9
Slenochironomus sp. 25.6 1.7 2.1 0.1 53.8 24
fanylarsus sp 10.4 0.8 5.1 0.3 12.5 0.6 7.7 0.3
Thienemannimyia series 29.2 2.3 69.2 45 31.3 1.6 115.4 5.2
Unid. Chironomidae pupa 91.7 7.2 89.7 5.8 1604 8.1 79.5 3~6
Mollusca
Gastropoda
Physidae
Physa sp. -- -- 4.2 0.2
Pelecypoda
Corbiculidae
Corbicula flummea -- 2.6 0.2 -- 2.6 01
Total Benthos 1,275.5 100 1,543.1 100 1,992.0 100 2,2327 100
Total Taxa"' 26 31 26 25
"There were highly significant differences between static ns|P = 0.0001). The number o< taxa at Station 3 was different than at Stations 1,
6, and 8 (P = 0.05I.
Note Total Taxa = distinct taxa; does not include pupa of included taxa
C-6
-------
Table C-5. Density (No./m2) and Percent Occurrence of Macroinvertebrates Collected at Stations 7 and 8 in the Ohio River,
Wheeling, West Virgina, July-August 1984
Station 7
Station 8
0.6 m
1.5 m
0.6 m
1.5 m
Taxa
Mean Mean
No./m2 Percent No./rnJ Percent
Mean Mean
No/m2 Percent No/mj
Percent
Nematoda 8.3 0.2
Platyhelminthes
Planarndae
Dugesia sp.
Annelida
Oligochaeta
Naididae
Unid. Naididae
3,229.2 81.3 297.3 25.3
Crustacea
Amphipoda
Gammaridae
Gammarus sp. 152.1
Acari
Hydracarina
I nsecta
Ephemeroptera
Heptageniidae
Stenonema sp.
S. femoratum
S. integrum
S. terminatum 4.2
Immature Heptageniidae
Caenidae
Caenis sp.
Baetidae
Baetis sp.
Trichoptera
Hydropsychidae
Cheumatopsyche sp.
Hydropsyche sp.
H. orris
H. orris pupa
H. valanis
Potamyia flava
Polycentropodidae
Cyrnetlus fraternus
Neureclipsis sp.
Coleoptera
Elmidae
Stenelmis sp. adult 2.1 <0.1
Diptera
Ceratopogonidae
Unid. Ceratopogonidae
Empididae
Unid. Empididae 2.1 <0.1
Chironomidae
Cricotopus bicinctus group 33.3 0.8
C. cylindraceus group 43.8 1.1
C. imersectus group 8.3 0.2
C. tremulus group 41.7 1.0
Dicrotendipes sp. 16.7 0.4
Micropsectra sp.
Nanocladius sp. 133.3 3.4
Parametr/ocnemus sp.
3.8 376.8 32.1
5.1 0.4
26 0.2
2.6 0.2
0.1
2.6 0.2
6.3
2.1
0.2
<0.1
5.1
74.3
7.7
2.6
5.1
0.4
6.3
0.7
0.2
0.4
5.1
2.6
0.4
0.2
8375
368.8
6.3
6.3
6.3
28.4
12.5
0.2
0.2
3.1
3.1
6.3
12.5
6.3
15.6
0.1
0.1
0.2
0.4
0.2
0.5
0.2
11.5
1384
615.2
7.7
3.8
3.8
69.2
77
346
7.7
3.8
0.8
9.1
40.6
0.5
0.3
0.3
4.6
0.5
2.3
0.5
0.3
17.9
35.9
20.5
38.5
7.7
33.3
5.1
1.5
3.1
1.7
3.3
0.7
2.8
04
40.6
218.8
37.5
159.4
262.5
43.8
181.3
1.4
7.4
1.3
5.4
8.9
1.5
6.1
~
3.8
19.2
11.5
15.4
50.0
30.8
19.2
7.7
03
1.3
0.8
1.0
3.3
2.0
1.3
0.5
C-7
-------
Table C-5. (continued)
Station 7
Station
06m
1 5 rr
0 6 in
1 5 m
Taxa
Paratanytarsus sp
Phaenopsectra sp
Polypedi/um conjictum type
P fal/ax group
P. scalaenum type
Pseudochironomus sp
Rheotanytarsus sp
Stenochironomus sp
Tanytarsus sp
Th/enemannimy/a series
Unid Ciironomidae pupa
Mollusca
Gastropoda
Physidae
Physa sp
Pelecypoda
Corbiculidae
Corbicula fluminea
Total Benthos
Total Taxa'*
Mean
Nc m- Percent
--
2.1 vO.I
188 05
2.1 '-.0.1
25.0 0.6
-
- -
_.
...
1563 39
833 2.1
2 1 <0 1
3,973.2 100
19
Mean
No rn
--
17.9
5.1
308
-
5.1
333
--
89.7
282
12.8
2.6
1,173.9
27
Percent
--
-
1.5
04
2.6
-
04
2.8
--
7.6
2.4
1.1
0.2
100
Mean
No TV
18.8
2344
53.1
3.1
78 1
-
12.5
1406
193.8
--
--
2,950.4
25
Percent
0.6
_..
7.9
1.8
0.1
2 6
-
0.4
48
6.6
--
--
100
Mean
No rn-
3 8
-
423
-
142.3
11 5
69.2
3.8
76 9
103 8
-
1,514.6
26
percent
03
2.8
-.
94
--
0.8
4.6
0.3
51
6.9
-
--
100
'"There were highly sign if icant differences between stations (P = 0.001). The number of taxa at Staion 3 was different than at Stations 1,6.
and 8{P = 0.05).
Note: Total Taxa = distinct taxa, does not include pupa of included taxa.
Table C-6. Numbers of Macroinvertebrates tor Each Replicate Sample Collected at Stations 1 and 2 in the Ohio River, Wheeling,
West Virginia, July-August 1984
Station 1
Station 2
Taxa
0.6 m
1.5 m
0.6 m
1.5 m
ABCABCAB C": A B C"
Coelenterata
Hydra sp
Platyhelmmthes
Planarndae
Dugesia sp
Annelida
Oligochaeta
Naididae
Und. Naididae
Crustacea
Amphipoda
Gammaridae
Gammarus sp
Acari
Hydracanna
Insecta
Ephemeroptera
Heptageniidae
Stenonema sp.
S integrum
S termmatum
11 15 6 2 7 10
17 18 37 29 15 10 120 249
105 120 178 260 161 157 40 205
42 98
158 56
C-8
-------
Table C-6. (continued)
Station ]
06m
Taxa A 3 C A
Caemdae
Caenis sp. - - 1
Odonata
Libellulidae
Perithemis sp.
Trichoptera
Hydropsychidae
Hydropsyche orris 5 1 2 --
H. simulans - - - - 1
Polycentropodidae
Cymel/us f rater nus - - 1 2 1
Neurec/ips/s sp
Diptera
Empididae
Unid. Emp.'didae -- 2 1
Chironomidae
Chironomus sp.
Cricotopus bicinctus group 8 2 6
C. cy/indraceus group 10 4 24
C intersectus group - 1
C. Iremulus group 12 6 12 1
Dicrotendipes sp. 22 18 6 4
Harnischia sp.
Micropsectra sp 12 6 2 3
M curvicornis - -- -- ~\
Nanoc/adius sp. 12 14 10
Parametriocnemus sp
Paratanytarsus sp ---21
Polypedilum convictum
type 16 14 10 2
P. /a//ax group - - -- 1
(ฐ scalaenum type - - 2 -- 4
Pseudoch/ronomus sp.
Rheotanytarsus sp. 88 108 124 4
Stenochironomus sp. - - 3
Tanytars us sp.
Thienemannimyia series 2 2
Unid. Chironomidae pupa 37 20 40 6
Mollusca
Gastropoda
Ancylidae
f err/ssia sp.
Physidae
Physa sp - - 2
Pelecypoda
Corbiculidae
Corbicula fluminea - - 1 0
Total Number of Taxa'b' 14 20 17 17
1 5 m
e
1
__
4
1
-
1
--
--
10
2
1
7
2
2
2
--
3
__
9
7
1
14
--
--
10
21
Statior 2
06m 15m
CAB C"1 A B C""
-
_-
4
1
10 - - - - 1
1 ... .- - i
2
-_
111-
3 2 15 1
3 2
4 2 ------
10 11 5 1 1
-.
3 i ---_
... __ .._
523-- 62--
3
1
3 3 35 22
1
2 - -- 6
2
49
3 - - 2 10
2 .
6 16 6 -- 1 8
32 3 4 1 2
1
__
6 - 1 -- - 1 -
23 13 13 9 12
'"One replicate substrate was not recovered.
'c'Total taxa values are for distinct taxa and do not include pupa.
C-9
-------
Table C-7. Numbers of Macroinvertebrates for Each Replicate Sample Collected at Stations 3 and 4 in the Ohio River, Wheeling,
West Virginia, July-August 1984
Station 3
Station 4
Taxa
Platyhelminthes
Plananaidae
0.6 m
ABC
1
1.5 m
ABC
0.6 m
ABC
1.5 m
A B
C
Annelida
Oligochaeta
Naididae
Unid Naididae
Crustacea
Amphipoda
Gammaridae
Gammarus sp
Decapods
Astacidae
Immature Astacidae
24
108
10 109
85
160
16
62
55
44
276
116
41
169
87
87
48
62
36
38
10
Acari
Hydracarina
Insecta
Ephemeroptera
Baetidae
Baetis sp
Heptageniidae
Stenacron interpunctatttm
Stenonema integrum
Immature Heptageniidae
Caenidae
Tricorythodes sp
Trichoptera
Hydropsychidae
Hydropsyche orris
Polycentropodidae
Cyrnellus fraternus
Neureciipsis sp.
Diptera
Empididae
Unid. Empididae
Chironomidae
Ablabesmyia sp.
Cricotopus bicinctus group
C cylindraceus group
C. intersectus group
C tremulus group
Dicrotendipes sp.
Endochironomus sp.
Glyptotendipes sp
Micropsectra sp.
Nanocladius sp.
Orthocladius sp.
Parach/ronomus sp
Parametriocnemus sp
Paratanytarsus sp
Phaenopsectra sp.
Polypedi/um convictum type
P. fa/lax group
P. scalaenum type
Pseudochironomus sp
Rheotanytarsus sp.
Stenochironomus sp.
Tanytarsus sp.
T-
..
13
"
__
7
1
2
43
__
1
--
15
1
-_
1
2
i
i
4
1 - 7 2
2
._
1
^ 1
4 __. ]
4 __ - _. 1
2 21 23 17 14 15
1
2
_
1
1
1
2 1 19
3 2 5 1
2
3
1
1
1
1
7
5
2
24
3
9
3
11
1
3
1
2
4
1
13
3
20
5
1
1
2
1
23
5
__
1
4 1
3 1
1
1
~_
1 1
15 13
2
1 2
1
3
1
9 23
5C
3
1
4
4
2
1
1
1
2
6
1
2
4
*
6
C-10
-------
Table C-7. (continued)
Station 3
0.6 m 1.5 m
Taxa A 8 C A B
Thienemannimyia series 41 31
Unid. Chironorridae pupae 213 2
Mollusca
Pelecypoda
Corbiculidae
Corbicula fluminea 36
Total Number of Taxa1" 11 5 10 11 5
Station 4
06 m
C A B C
6477
2 6 15 20
7
9 20 18 15
1.5 m
ABC
5133
387
11 19 19
Table C-8. Numbers of Macrotnvertebrates for Each Replicate Sample Collected at Stations 5 and 6 in the Ohio River, Wheeling,
West Virginia, July-August 1984
Station 5
Station 6
0.6 m
1.5 m
0.6 m
Taxa
B
B
B
1.5 m
B
Platyhelminthes
Planariidae
Dugesia sp
Ectoprocta
Plumatellidae
Hyalinel/a punctata
Annelida
Oligochaeta
Naididae
Unid. Naididae
Crustacea
Amphipoda
Gammaridae
Gammarus sp.
Acari
Hydracarina
Inseeta
Epherneroptera
Heptageniidae
Stenacron interpunctatum
Stenonema integrum
Odonata
Libelluiidae
Perishemis sp.
Trichoptera
Hydropsychidae
Cheumatopsyche sp.
Hydropsyche orris
H. orris pupa
H. simulans
Potamyia f/ava
Symphitopsyche morosa
Polycentropodidae
Cyrnellus fraternus
Neureclipsis sp.
Leptoceridae
Oecetis sp
47
16
36
38
39
2 1
6 11 4 227 38 4 43
66 56 58 17 66 160 153
5 1 2 1
17 6
71 109
9
2
7
1
1
1
1
11 20 7
1
1
3 5 23
1 1
2
1
3
14 20 1
1
33 13
1
2
12
7
^
1
1
3
1
3 28
4.
T
2 6
6
1
1
17
3
6
1
27
2
C-11
-------
Table C-8 (continued)
Station 5
Taxa
Diptera
Empididae
Unid. Empididae
Chironomidae
Ablabesmyia sp.
Cncotopus bicincws group
C. cy/indraceus group
C intersectus group
C. tremulus group
Dicrotendipes sp
Glyptotendipes sp.
Micropsectra sp.
M curvicornis
Microtendipes sp.
Nanocladius sp
Parametriocnemus sp
Paratanytarsus sp
Phaenopsectra sp
Polypedilum convictum type
P. la/lax group
P scalaenum sp.
Pseudochironomus sp.
Rheotanytarsus sp.
Stenochironomus sp.
Tanytarsus sp.
Thienemann/my/a series
Unid Chironomidae pupa
A
---
--
2
70
10
8
18
--
_..
--
14
--
2
-
12
--
2
--
10
-
2
2
15
06m
B
-
-
1
2
7
11
11
--
1
1
--
15
--
--
1
5
--
3
1
4
--
3
6
11
C
--
--
30
10
7
12
3
-
1
8
--
1
1
8
--
1
7
--
--
6
18
A
--
1
--
12
2
4
18
1
--
6
1 1
T
1
-
3
-
5
-
6
4
2
5
11
1.5 m
B
1
1
--
7
3
2
11
1
---
2
2
--
-
-
18
--
12
1
3
1
---
16
15
C
1
--
8
2
1
24
--
-
5
3
--
--
7
--
4
5
5
6
9
Molluscs
Gastropoda
Physidae
Physa sp
Pelecypoda
Corbiculidae
Corbicula fluminea
Total Number of Taxa""
16
21
20
1
26
24
11
7
Station 6
0 6 rn
B
18
16
22
28
50
2
32
1
52
10
13
3
1
14
19
1
1
27
5
4
18
4
6
4
18
18
6
12
14
20
14
32
11
19
14
19
20
18
1 5 m
B
5
3
3
10
14
7
33
6
10
10
2
1
3
12
12
1
33
1
3
3
10
20
1
20
""Colonial organisms present, not included in total taxa count.
'"'Total taxa values are for distinct taxa and do not include pupa.
C-12
-------
Table C-9 Numbers of Macroin vertebrates for Each Replicate Sample Collected at Stations 7 and 8 in the Ohio River, Wheeling,
West Virginia, July-August 1984
Station 7
Station 8
Taxa
Nematoda
Platyhelminthes
Planariidae
Dugesia sp.
0.6 m 1.5 m 0.6 m 1.5 m
ABCABCAB C1" A B C1"
1 3
Annelida
ONgocnaeta
Naididae
Unid. Naididae 612
Crustacea
Amphipoda
Gammaridae
Gammarus sp. 21
Acari
Hydracarina
Insecta
Ephemeroptera
Heptageniidae
Stenonema sp.
S. femoratum
S. integrum
S. terminatum 1
Immature Heptageniidae
Caenidae
Caenis sp
Baetidae
Baetis sp.
Trichoptera
Hydropsychidae
Cheumatopsyche sp.
Hydropsyche sp
H. Orris
H. orris pupa
H. valanis
Potamyia f/ava
Polycentropodidae
Cyrne/lus fraternus
Neurec/ipsis sp
Coleoptera
Elmidae
Stenelmis sp. adult
Diptera
Ceratopogonidae
Unid. Ceratopogonidae
Empididae
Unid. Empididae
Chironomidae
Cricotopus bicinctus group
C. cyt/ndraceus group
C intersectus group
C. tremulus group
Dicrotendipes sp
Micropsectra sp.
Nanocladius sp.
Parametriccnernus sp.
Paratanytarsus sp.
Phaenopsectra sp. 1
Polypedilum convictum
type 3
265 673
16
36
49 33 34 193
__
1
1
--
1
1
4
10
7
2
20
1
1
3
2
1
5
3
22
__
1
--
--
9
9
3
8
3
22
14
1
1
1
11
4
6
2
6
6
1
1
2
2
2
2
a
1
3
2
9
2
--
1
4
1
2
1
--
4
2
3
_-
1
2
12
50
12
38
76
10
40
R
77
35 50 62 98 20
58
1
20
13
8
4
18
17
18
96
3
8
5
3
1
1
18
64
11
1
2
3
1
5
3
2
1
C-13
-------
Table C-9. (continued)
Station 7 Station 8
06m
Taxa
A
1
9
__
-
--
-
41
16
06m
B
2
_-
-
.-
15
5
C
_
1
--
--
--
19
19
A
2
6
--
1
12
--
8
3
1 5 m
B
_.
4
-
--
--
17
5
C
__
2
--
1
1
10
3
A
__
12
10
-
2
32
41
06m
B C1"
__
5
1
15
2
13
21
A
28
-
1
12
--
13
13
1 b
3
__
9
__
2
6
1
7
14
P faltax group
P scalaenum type
Pseudochironomus sp
Rheotanytarsus sp.
Stenochironomus sp.
fanytarsus sp
Jhienemannimyia series
Unid. Chironomidae pupa
Mollusca
Gastropoda
Physidae
Physa sp. 1 - - 1 2 2 ----- -
Pelecypoda .
Corbiculidae
Corbicula f/uminea .__-_. 1 _ _ .
Total Number of Taxa'" 18 14 13 20 17 20 22 19 - 19 23
""One replicaie substrate was not recovered.
'"Total taxa values are for distinct taxa and do not include pupa.
C-14
-------
Table C-10. Analysis of Variance and Tukey's Studentized
Range Test for Zooplankton, Ohio River"
Crustaceans
Dependent variable: In count
Sum of
Source df Squares
Model
Error
Corrected total
Station
Depth
Station
Mean
8
7
15
7
8
Tukey'
3
5.95
Mean
Square F Value
27.89 349 4.68
522 0.74
33.11
2305 4.42
4.84 6.49
s Studentized Range Test
52487
5.20 3.95 3 70 3.55 3.40 3
PR >F
0.0283
0.0343
0.0382
6 1
.10 1.75
Rotifers
Dependent variable: count
Sum of
Source df Squares
Model
Error
Corrected total
Station
Depth
Station
Mean
8
7
15
7
1
Tukey'
8
4.82
Mean
Square F Value
PR > F
3.59 0.45 11.3 0.0022
027 0.04
3.87
359 12.90 0.0016
0.004 0.11 0.7535
s Studenti2ed Range Test
6473521
4.63 4.63 4.62 4.57 4.50 4.44 3.20
TotalZooplankton
Dependent variable: In count
Sum of
Source df Squares
Model
Error
Corrected total
Station
Depth
Station
Mean
Mean
Square F Value
8 3.48 0.43 11.17
7 0.27 0.04
15 375
7 347 12.73
1 0.008 0.23
Tukey's Studentized Range Test
846735
4.85 4.67 4.67 4.65 4.63 4.56 4
PR >F
0.0023
0.0017
0.6496
2 1
.48 3.27
"'SASPROCGLM.
Table C-1 1 . Analysis of Variance and Confidence Interval-
Overlap Results of Clorophyll a and Biomass
Measurements of Periphyton, Ohio River '"
Chlorophyll a
Dependent variable Chla (all stati
Sum of
Source df Squares
Station
Error
Corrected total
95
Station
Mean
ons|
Mean
Square F Value
5 26,104 5,221 648
9 7,250 806
14 33,354
Percent Confidence Interval Overlap
23476
40.1 31.2 29.1 731 122.5
Dependent variable: Chla (Station
Sum of
Source df Squares
Station
Error
Corrected total
95
Station
Mean
4
9
13
Percent
2
40.1
SI-/)
Mean
Square F Value
18,371 4,593 570
7,250 806
25,621
Confidence Interval Overlap
3476
31.2 29.1 73.1 122.5
Biomass
Dependent variable: In AFDW (all stations)
Sum of Mean
Source df Squares Square F Value
Station
Error
Corrected total
95
Station
Mean
5
9
14
Percent
3
1.26
312 062 370
1.52 0.17
4.64
Confidence Interval Overlap
4278
1.14 1.70 169 2.41
Dependent variable: In AFDW (Stations 1 -7)
Sum of Mean
Source df Squares Square F Value
Station
Error
Corrected total
95
Station
Mean
4
9
13
Percent
3
1.27
2.54 064 3.76
1.52 0.17
406
Confidence Interval Overlap
4276
1.14 1 70 1 69 2.32
PR >F
0.008
a
151.6
PR > F
0.014
PR >F
0.043
6
2.32
PR >F
0.046
""MINITAB.
C-15
-------
Table C-1 2 Analysis of Variance and Tukey's Studentized Range Test Results for Oligochaetes and Amphipods, Ohio River
Oligochaete (unidentified Naididae)
Dependent variable counts
Source
Model
Error
Corrected total
Station
Depth
Station Depth
Station
Mean
Dependent variable: counts
Source
Model
Error
Corrected total
Station
Depth
Station Depth
Station
Mean
df
15
28
43
7
1
7
7
277.67
df
15
28
43
7
1
7
1
163.50
Sum of Squares Mean Square F Value
682,986.22 45,532.41 6.38
199,858.50 7,13780
882,844.72
292,953.14 586
104.50000 14.64
285,561.81 572
Tukey's Studentized Range Test
238461
127.25 8167 76.50 6617 5583 21.00
Amphipod (Gammarus sp.)
Sum of Squares Mean Square F Value
92.800.56 6,186.70 2.97
58.23116 2,07968
151,031.72
81,327.47 559
1,530.67 0.74
9,66572 066
Tukey's Studen'ized Range Test
263857
11475 9600 9583 6950 5033 36.67
PR > F
0.0001
0.0003
0.0007
00004
5
12.50
PR > F
00062
00004
03982
0 7002
4
30.50
Table C-1 3. Analysis of Variance and Tukey's Studentized Range Test Results for Chironomidae Taxa, Ohio River
All Chironomid Taxa
Dependent variable: counts
Source
df
Sum of Squares
Mean Square
F Value
PR
Model
Error
Corrected total
Station
Depth
Station Depth
Station
Mean
15
28
43
7
1
7
8
174.00
166,349.01
69,444.17
235,793.18
90,264.93
44,118.37
36,723.72
Tukey's Studentized Range Test
1 6 5
14450 12817 105.00
11,08993
2,480.15
7
6917
447
5.20
17.79
212
4 2
6917 4175
00003
00007
00002
00750
3
3250
C-76
-------
Table C-13. (continued)
Dependent variable. In counts
Source
Model
Error
Corrected total
Station
Depth
Station Depth
Station
Mean
Dependent variable: In count
Source
Model
Error
Corrected total
Station
Depth
Station Depth
Station
Mean
Dependent variable, counts
Source
Model
Error
Corrected total
Station
Depth
Station Depth
Station
Mean
df
15
28
43
7
1
7
3
2.79
df
15
28
43
7
1
7
8
2 64
df
15
28
43
7
1
7
1
6367
Dicrotendipes sp.
Sum of Squares Mean Square F Value
1717 1.14 2.79
11.51 041
28.68
12.22 4.25
T.50 3.65
4.06 1.41
Tukey's Studentized Range Test
54861 7
2.77 2.73 263 2.62 240 1.45
Polypedilum conviction type
Sum of Squares Mean Square F Value
32.07 2.14 3.79
15.79 0.56
47.86
16.33 4.14
12.05 21.38
4.30 1.09
Tukey's Studentized Range Test
654127
2.60 2.16 1.94 1.92 1.79 1.18
Rheoianytarsus sp.
Sum of Squares Mean Square F Value
31,47913 2,098.61 2248
2,61367 93.34
34,092.80
20,129.63 30.81
1,390.29 14.89
9.77748 14.96
Tukey's Studentized Range Test
685473
24.17 7.00 5.83 0.83 0.33 0.00
PR >F
0.0093
00026
00664
0.2399
2
1.41
PR >F
0.0011
0.0031
00001
03955
3
0.76
PR > F
0.0001
0.0001
0.0006
0.0001
2
0.00
C-17
-------
Table C-13.
(continued)
Unidentified Chironomidae Pupae
Dependent variable counts
Source
Model
Error
Corrected total
Station
Depth
Station Depth
Station
Mean
Dependent variable: In count
Source
Model
Error
Corrected total
Station
Depth
Station Depth
Station
Mean
d!
15
28
43
7
1
7
1
24.83
df
15
28
43
7
1
7
5
2.61
Surr of Squares Mean Square F Value
3.96226 26415 342
2.164.17 7729
6,12663
2,71935 5.03
836.23 1082
39635 073
Tukey's Studentized Range Test
865472
2225 1800 1317 983 850 250
Cncotopus cylindraceus
Sjm of Squares Mean Square F Value
37.33 2.49 3 90
17.86 064
55 19
1855 4.15
1588 2490
4.06 091
Tukey's Sludentized Range Test
8671 24
236 1.91 1 68 1.43 1 14 092
PR - F
00024
0.0009
00027
0.6462
3
1.67
PR -F
0.0009
00030
00001
0.5141
3
061
C-J8
-------
Table C 14 Analysis of
Dependent variable count
Source
Model
Error
Corrected total
Station
Depth
Station Depth
Station
Mean
Dependent variable: In count
Source
Model
Error
Corrected total
Station
Depth
Station Depth
Station
Mean
Table C- 15 Analysis of
River
Dependent variable:
Source
Model
Error
Corrected total
Station
Depth
Statton
Mean
Variance and
df
15
28
43
7
1
7
5
13.50
df
15
28
43
7
1
7
5
2.43
Variance and
df
8
35
43
7
1
5
21.17
T ukey's Studentrzed Range Test Results for Trichoptera, Ohio R iver
Hydropsyche orris
Sum of Squares Mean Square F Value
1,060.97 70.73 279
708.67 2531
1,769.64
836.30 4.72
73.50 2.90
153.39 0.87
Tukey's Studentized Range Test
68741 3
8.50 5.50 5.33 3.00 2.00 0.00
Cyrneltus fraternus
Sum of Squares Mean Square F Value
36.76 2.45 5.22
13.15 047
49.91
23.13 7.03
813 17.30
4.72 1.44
Tukey's Studentized Range Test
683147
1.66 1.30 1.08 1.08 0.53 018
Tukey's Studentized Range Test Results for the Benthic Macroinvertebrate
Sum of Squares Mean Square F Value
76462 95.58 12.58
266.02 7.60
1,030.64
746.80 14.04
1782 234
Tukey's Studemized Range Test
81 6472
20.75 18.67 18.50 17.00 16.33 11.75
PR > F
0.0091
0.0013
0.0994
0.5449
2
0.00
PR > F
0.0001
0.0001
00003
0.2310
2
0.17
Taxa, Ohio
PR >F
0.0001
0.0001
01347
3
833
C-19
& U. S. GOVERNMENT PRINTING OFFICE 1986 ''646-116 '20792
------- |