c/EPA
United States
Environmental Protection
Agency
Environmental Research
Laboratory
Duluth MN 55804
EPA/600/8-86/002
March 1986
Research and Development
Validity of
Effluent and Ambient
Toxicity Tests for
Predicting Biological
Impact,
Skeleton Creek,
Enid, Oklahoma
-------
EPA/600/8-86/002
March 1986
Validity of Effluent and Ambient
Toxicity Tests for Predicting
Biological Impact,
Skeleton Creek, Enid, Oklahoma
Edited by
Teresa J. Norberg-Kmg and Donald I Mount
Environmental Research Laboratory
6201 Congdon Blvd
Duluth, Minnesota 55804
Environmental Research Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Duluth, MINI 55804
U.S. Environmental Protection Agency
Region 5, Library (PL-12J)
77 West Jackson Boulevacd, 12th Ftoor
Chicago, IL 60604-3590
-------
Notice
This document has been reviewed in accordance with U.S. Environmental
Protection Agency policy and approved for publication. Mention of trade names
or commercial products does not constitute endorsement or recommendation
for use.
-------
Foreword
The Complex Effluent Toxicity Testing Program was initiated to support the
developing trend toward water quality-based toxicity control in the National
Pollutant Discharge Elimination System (NPDES) permit program. It is designed
to investigate, under actual discharge situations, the appropriateness and utility
of "whole effluent toxicity" testing in the identification, analysis, and control of
adverse water quality impact caused by the discharge of toxic effluents.
The four objectives of the Complex Effluent Testing Program are:
1. To investigate the validity of effluent toxicity tests in predicting adverse
impact on receiving waters caused by the discharge of toxic effluents.
2. To determine appropriate testing procedures which will support regulatory
agencies as they begin to establish water quality-based toxicity control
programs.
3. To provide practical case examples of how such testing procedures can be
applied to effluents discharged to a receiving water.
4. To field test short-term chronic toxicity tests involving the test organisms,
Ceriodaphnia 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 the controls largely
in place, future controls for toxic pollutants will, of necessity, be based on
site-specific water quality considerations.
Setting water quality-based controls for toxicity can be accomplished in two
ways. The first is the pollutant-specific approach which involves setting limits for
single chemicals, based on laboratory-derived no-effect levels. The second is the
"whole effluent" approach which involves setting limits using effluent toxicity
as a control parameter. There are advantages and disadvantages to both
approaches.
The "whole effluent" approach eliminates the need to specify a limit for each of
thousands of substances that may be found in an effluent. It also includes all
interactions between constituents as well as biological availability. Such limits
determined on fresh eflfuent may not reflect toxicity after aging in the stream
and fate processes change effluent composition. This problem is less important
since permit limits are normally applied at the edge of the mixing zone where
aging has not yet occurred.
To date, eight sites involving municipal and industrial dischargers have been
investigated. They are, in order of investigation:
-------
1. Scippo Creek, Circleville, Ohio
2. Ottawa River, Lima, Ohio
3. Five Mile Creek, Birmingham, Alabama
4. Skeleton Creek, Enid, Oklahoma
5. Naugatuck River, Waterbury, Connecticut
6. Back River, Baltimore Harbor, Maryland
7. Ohio River, Wheeling, West Virginia
8. Kanawha River, Charleston, West Virginia
This report presents the site study on Skeleton Creek, Enid, Oklahoma, which
was conducted in August 1983. The stream is small and receives discharges
from two industries and one publicly owned treatment works.
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
IV
-------
Contents
Page
Foreword iii
List of Figures vii
List of Tables viii
Acknowledgments x
List of Contributors xi
Executive Summary xii
Quality Assurance xiii
1. Introduction 1-1
2. Study Design and Site Description 2-1
3. Hydrology Survey 3-1
3.1 Discharge Flow Measurements 3-1
3.2 Flow Contribution 3-2
4. Laboratory Toxicity Tests 4-1
4.1 Chemical and Physical Test Conditions 4-1
4.2 Effluent Toxicity Test Results 4-1
4.3 Ambient Toxicity Test Results 4-2
4.4 Discussion 4-3
5. Plankton Community Survey 5-1
5.1 Community Structure 5-1
5.2 Evaluation of the Plankton Community 5-1
6. Macroinvertebrate Community Survey 6-1
6.1 Community Composition 6-1
6.1.1 Natural Substrates 6-1
6.1.2 Artificial Substrates 6-1
6.1.3 Comparison Between Substrate Type 6-1
6.2 Station Comparisons 6-2
6.2.1 Natural Substrates 6-2
6.2.2 Artificial Substrates 6-4
6.2.3 Gear Comparison 6-4
6.3 Evaluation of the Macroinvertebrate Community 6-4
-------
Contents (cont'd)
Page
7. Fish Community Survey 7-1
7.1 Community Structure 7-1
7.2 Evaluation of the Fish Community 7-1
8. Comparison Between Laboratory Toxicity Tests and
Instream Biological Response 8-1
8.0 Background 8-1
8.1 Prediction of Instream Community Impacts Based on
Effluent Dilution Test Results 8-2
8.2 Comparison of Ambient Toxicity Test Results and Field Data .... 8-3
8.3 Summary 8-4
References R-1
Appendix A: Hydrological Sampling and Analytical Methods A-1
Appendix B: Toxicity Test and Analytical Methods B-1
Appendix C: Biological Sampling and Analytical Methods C-1
Appendix D: Biological Data D-1
-------
List of Figures
Number Title Page
2-1 Map of study site on Skeleton Creek, Enid, Oklahoma 2-3
5-1 Densities of crustaceans, rotifers, and algae at Skeleton
Creek, and Boggy Creek, Enid, Oklahoma, August 1983 5-2
6-1 Mean densities of Dicrotendipes sp. collected from Skeleton
Creek and Boggy Creek, Enid, Oklahoma, August 1983 6-3
6-2 Mean densities of Berosus sp. collected from Skeleton Creek
and Boggy Creek, Enid, Oklahoma, August 1983 6-3
8-1 A comparison of percent toxicity and percent reduction
of the taxa 8-5
VII
-------
List of Tables
Number Title Page
2-1 Sample Collections Conducted for the Quantitative Biological
Assessment and Ambient Toxicity Tests, Skeleton Creek
and Boggy Creek, Enid, Oklahoma, August 1983 2-1
3-1 Measured Flows and Discharges on Skeleton Creek and
Boggy Creek, Enid, Oklahoma 3-1
3-2 Effluent Pumping Records and Daily Average Discharge at the
Fertilizer Plant, Enid, Oklahoma 3-1
3-3 Mean Flow and Percent Flow Contribution from Three
Discharges for Boggy Creek and Skeleton Creek, Enid,
Oklahoma, August 1983 3-2
4-1 Routine Chemistry Data for Effluent and Ambient Tests 4-3
4-2 Final Dissolved Oxygen and Final pH for Ceriodaphnia
Effluent and Ambient Toxicity Tests 4-3
4-3 Seven-Day Survival of Larval Fathead Minnows in Two
Effluents 4-3
4-4 Mean Individual Dry Weight (mg) After Seven Days for Larval
Fathead Minnows Exposed to Two Effluents 4-4
4-5 Percent Survival and Young Production of Ceriodaphnia in
Two Effluents 4-4
4-6 Seven-Day Percent Survival of Larval Fathead Minnows in
the Ambient Toxicity Test 4-4
4-7 Mean Individual Dry Weight (mg) After Seven Days for Larval
Fathead Minnows in the Ambient Toxicity Test 4-5
4-8 Percent Survival and Mean Young Production of Ceriodaphnia
in the Ambient Toxicity Test 4-5
5-1 Mean Density (Number/Liter) of Planktonic Organisms
Collected in Skeleton Creek and Boggy Creek, Enid,
Oklahoma, August 1983 5-1
6-1 Mean Percent Composition of Major Macroinvertebrate Taxa
Collected from Natural Substrates in Skeleton Creek and Boggy
Creek, Enid, Oklahoma, August 1983 6-2
6-2 Mean Percent Composition of Major Macroinvertebrate Taxa
Collected from Artificial Substrates in Skeleton Creek and
Boggy Creek, Enid, Oklahoma, August 1983 6-3
-------
List of Tables (cont'd)
Number Title Page
6-3 Total Taxa Collected by Artificial and Natural Substrates and
the Combined Macromvertebrate Taxa at Each Sampling
Station 6-5
7-1 Number of Fish Collected by Seine from Skeleton and Boggy
Creeks, Enid, Oklahoma, August 1983 7-1
8-1 Comparison Between the Acceptable Effluent Concentration
(AEC) and the Instream Waste Concentration (IWC) for
Effluents Tested 8-3
8-2 Percent Increase in Toxicity and Percent Reduction in Number
of Taxa for the Instream Biological Community 8-3
8-3 Percent of Correct Predictions Using Four Levels of Defined
Impact 8-4
C-1 Station Description Information and Estimated Proportions
of Riffle and Pool for Skeleton Creek and Boggy Creek, Enid,
Oklahoma C-1
D-1 Mean Density (No./m2) of Benthic Macroinvertebrates
Collected from Natural Substrates in Skeleton Creek and Boggy
Creek, Enid, Oklahoma, August 1983 D-1
D-2 Mean Densities (No./m2) of Macroinvertebrates
Collected from Artificial Substrates in Skeleton Creek and
Boggy Creek, Enid, Oklahoma, August 1 983 D-2
D-3 Analysis of Variance and Tukey's Studentized Range Test
Results for Zooplankton, Skeleton Creek, August 1983 D-3
D-4 Analysis of variance and Tukey's Studentized Range Test
Results for the Two Most Abundant Macroinvertebrate Taxa
from the Natural Substrates, Skeleton Creek, August 1983 D-4
D-5 Analysis of Variance and Tukey's Studentized Range Test
Results for the Two Most Abundant Macroinvertebrate Taxa
from the Artificial Substrates, Skeleton Creek, August 1 983 .... D-5
D-6 Analysis of Variance and Tukey's Studentized Range Test
Results for Numbers of Macroinvertebrate Taxa,
Skeleton Creek, August 1983 D-5
D-7 List of Fish Species and Families Collected from Skeleton
Creek and Boggy Creek near Enid, Oklahoma D-6
IX
-------
Acknowledgements
The assistance of Floyd Boettcher, Environmental Research Laboratory-Duluth,
in collecting the effluent and ambient water samples as well as his help in the
biological survey work is acknowledged. The assistance of Ron Jarman from the
Oklahoma Water Resources Board, Oklahoma City, Oklahoma, in the site
selection is appreciated. The help of Allen Burton, EPA, Region 6, Dallas, Texas,
insetting the Hester-Dendy samples is also noted. Finally, many thanks to Terry
Highland for her assistance in the final typing.
-------
List of Contributors
LABORATORY TOXICITY TESTS
Teresa J. Norbert-King1 and Thomas H. Roush1
HYDROLOGY SURVEY
Tomas M. Farnam2 and Jonathan C. Yost2
PLANKTON COMMUNITY SURVEY
Thomas E. Roush1 and Michael T. Barbour2
MACROINVERTEBRATE COMMUNITY SURVEY
Thomas E. Roush1, Kimberly D. Juba2, and Sarah G. Wood2
FISH COMMUNITY SURVEY
Michael T. Barbour2, Thomas E. Roush1, and Kimberly D. Juba2
COMPARISON OF LABORATORY TOXICITY TEST DATA AND
RECEIVING WATER BIOLOGICAL IMPACT
Teresa J. Norbert-King1 and Donald I. Mount1
PRINCIPAL INVESTIGATOR
Donald I. Mount, Ph.D.1
Environmental Research Laboratory—Duluth, U S Environmental Protection Agency, 6201 Congdon Blvd ,
Duluth, Minnesota 55804
2EA Engineering, Science, andTechnology, Inc , Hunt Valtey/Loveton Center, 1 5 Loveton Circle, Sparks, Maryland
21152
-------
Executive Summary
Skeleton Creek was studied in August 1983 and was the fourth site study.
Skeleton Creek is located in an agricultural area in northwestern Oklahoma,
near Enid. The creek has a shallow gradient with mostly sand and sandstone
bedrock. A small creek, Boggy Creek receives discharges from both an oil
refinery and a publicly owned treatment works (POTW) prior to its confluence
with Skeleton Creek. A fertilizer processing plant discharge is located on
Skeleton Creek just downstream of the confluence of the two streams.
The toxicity of two effluents and ambient stream stations were evaluated.
Hydrological and ecological field surveys were also done. A comparison of the
relationship between the measured toxicity of the water samples collected from
the stream and the health of the aquatic community at the same stream stations
is made.
The results of the toxicity tests found the fathead minnow 7-day growth test to be
more sensitive to both effluents than the 7-day Ceriodaphnia reproduction test.
Station 5, below all three discharges, was the station where the toxicity tests,
zooplankton and fish were the most affected.
Both the toxicity test data andtheecological survey data showthat impact at the
stream stations is correlated with the toxicity measured (number of species lost).
Correct predictions were made for 87.5 percent of the stations when any equal
level of impairment and toxicity was compared.
The results of this study combined with those previous studies published
(Mount et al., 1 984, and Mount and Norberg, 1985) and ones yet to be published
(i.e., Mount et al., 1 985) will be used to recommend the best available approach
to predict the impacts of discharges on biological communities using effluent
and ambient toxicity tests. The data from this study clearly indicate the utility of
effluent and ambient toxicity tests for predicting instream effects.
XII
-------
Quality Assurance
Coordination of the study was done by the principal investigator preceding any
field work or toxicity testing. A reconnaissance trip was made to the site in the
spring of 1983 to obtain the necessary details regarding each discharge and to
make a cursory evaluation of the stream. Following that trip, the details were
delineated for setting sampling dates and the specific sampling sites; and the
specific measurements to be made for each stream station. This study required
coordination in setting artificial substrates, removing the substrates, planning
the hydrological and ecological surveys, and collection of effluents and water
samples for the toxicity tests by two organizations (see list of contributors). The
principal investigator was responsible for all the quality assurance related
decisions. All instrumentation used during the study were calibrated daily
according to manufacturers specifications. Test organisms for the toxicity tests
were laboratory raised.
-------
7. Introduction
Future activities in water pollution control will focus,
in part, on the control of toxic pollutants that impact
water quality. There are two methods used in
controlling toxic impact: pollutant-specific controls
and "whole effluent toxicity" controls. Because
toxicity testing evaluates a living organism's re-
sponse, it has an advantage over chemical-specific
analyses which may not identify all pollutants in a
wastewater sample and which cannot detect toxicity
interactions. Toxicity information can provide a basis
for permit limits based on state water quality stan-
dards for toxicity- or technology-based requirements.
The primary purpose of this study is to investigate the
relationship between effluent and ambient toxicity,
and community response. Toxicity tests have the
potential to predict instream impact.
This report is organized into chapters corresponding
to the project tasks. Following an overview of the site
description and study design, the chapters are
arranged into hydrological survey chapters, toxicity
test results, and ecological survey results for the
study. An integration of the laboratory and field
studies are presented in Chapter 8. All the laboratory
methods, hydrology methods, ecological survey meth-
ods, and supporting data are presented in the
appendices.
1-1
-------
2. Study Design and Site Description
The study area was on Skeleton Creek, which
originates 6.4 km northeast of Enid, Oklahoma. Boggy
Creek begins 3 km south of Enid and flows southeast
for 1 2 km before its confluence with Skeleton Creek,
which then flows 105 km before its confluence with
the Cimarron River. A Refinery and a Publicly Owned
Treatment Works (POTW) discharge treated effluent
into Boggy Creek. The POTW is an activated sludge
plant. The most upstream discharge is the refinery,
but the POTW discharge is only 0.2 km downstream
of it. Little mixing occurs before the refinery effluent
meets the POTW outfall. A Fertilizer manufacturing
plant discharges its treated effluent into Skeleton
Creek 0.5 km downstream from the confluence with
Boggy Creek. The streams maintain a shallow gradi-
ent of 1 m/km. During the 1983 field sampling period,
the POTW pumped its treated wastewater at night to
both the Refinery and Fertilizer Plant for use as
process water and the POTW discharged directly into
Boggy Creek during the day. The Refinery discharged
continuously to Boggy Creek while the Fertilizer Plant
discharged intermittently into Skeleton Creek. Actual
discharge flow measurements are given in Chapter 3.
Study components include 7-day Ceriodaphnia3
reproductive toxicity tests and 7-day larval growth
tests on fathead minnows on ambient samples from
the stream stations and various concentrations of the
Refinery and Fertilizer Plant effluents. The POTW
effluent was not tested because during the study
period the plant anticipated that its discharge would
all go to the Refinery and Fertilizer Plant. Also, stream
flow and discharge volume measurements, quanti-
tative assessment of the planktonic, macroinverte-
brate, and fish communities were made. Artificial
substrates were set in the stream July 20 and were
removed when field sampling, effluent, and water
sampling was completed August 9 to 11,1983. Water
samples for the toxicity tests were collected at
locations near where the artificial and natural
substrate samples were taken. The toxicity tests were
conducted August 14 to 21 at the Environmental
'The species of Cenodaphnia used for this study is not known with certainty
The stock cultures were earlier identified as C reticu/ata but in November
1983, based on taxonomic verification by Dorothy Berner, Ph D , Temple
University, PA, a second species C dubia was also identified in the stock
cultures The exact determination of the species tested is not critical to the
results of this study Therefore, all references to Cenodaphnia are to genus
level only
Research Laboratory-Duluth. Table 2-1 presents the
type of sampling done for e.ach stream station.
The study area on Skeleton Creek and Boggy Creek
covered a total of 26.6 river kilometers (RK). River
kilometers were estimated from county topographical
maps using the confluence with the Cimarron River
as zero river kilometers. The streams have been
described in reports by Wilhm (1 965), Baumgardner
(1966), and Namminga (1975). Both creeks are
shallow prairie streams with shallow tributaries
having low summer or intermittent flows. Pool areas
predominate with periodic riffles and runs along their
lengths. Twelve sampling stations are located along
the study area (Figure 2-1) and are described below.
The habitats sampled were pools, riffles, and runs for
the benthic macroinvertebrates, pools for the fish,
and moving water areas for the plankton. The
estimated cover, percent riffle, and percent pool for
each station is presented in Table C-1.
The station descriptions are as follows:
Station 1A (RK 2.8)—The uppermost point sampled
on Boggy Creek, which was 4.6-6.0 m wide at low
flow. The riffle was 0.1 5 m deep and the pool was
0.45 m deep. The sides were lined with riparian
vegetation which provided nearly 100 percent
cover. Macroinvertebrates were collected from the
small riffle areas created by flat rocks. Seining for
the fish survey was conducted in the adjoining
pool.
Station 1 (RK 1.2)—Upstream from the Refinery
discharge on Boggy Creek. The station was used
only for setting artificial substrates for sampling
macroinvertebrates. After the substrates were set,
construction of a beaver dam impounded water
and formed a turbid pool, 0.76 m deep. Riparian
vegetation on the shore provided 100 percent
cover.
Station 2 (RK 107.0)—On Skeleton Creek, 2 km
upstream of the confluence of Skeleton Creek and
Boggy Creek. Riparian vegetation on the shore
provided 90 percent cover. The station had a
sandy-bottom pool 0.76 m deep and a shallow run
less than 0.15 m deep with a bottom substrate
composed of sand and gravel.
Station 3 (RK 0.2)—On Boggy Creek below both the
POTW and Refinery discharges and just prior of the
2-1
-------
Table 2-1.
Sample Collections Conducted for the Quantitative Biological Assessment and Ambient Toxicity Tests, Skeleton
Creek and Boggy Creek, Enid. Oklahoma, August 1983
Collections
Sampling Stations
1A
5A
10
Plankton
Macroin vertebrates
Natural substrate
Artificial substrate
Fish
Ambient Toxicity Test
Fathead minnow
Cenodaphnia dubia
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X X
X
X
X
confluence with Skeleton Creek. The station was
composed of a 0.6 m deep pool with a sand bottom
and sand overlying rock, and a shallow run less
than 0.15 m deep. Riparian vegetation provided
about 80 percent cover.
Station 4 (RK 104.8)—Downstream of the con-
fluence of the creeks and upstream of the Fertilizer
Plant discharge. The run (0.3 m deep) substrate
was composed of irregular rock with a covering of
attached filamentous algae and sand. In addition, a
black, flocculent material had aggregated in a few
areas on the bottom. Riparian vegetation provided
no cover.
Station 5 (RK 104.3)—On Skeleton Creek, 0.3 km
downstream of the Fertilizer Plant. The water
contained large amounts of floating algae and a
black, flocculent material. The station was com-
posed of a 0.6 m deep pool and a 0.1 5 m deep riffle
area with a rocky bottom. Riparian vegetation
provided 10 percent cover.
Station 5A (RK 103.6)—At Southgate Road cross-
ing of Skeleton Creek. The station was composed
of a pool, 0.45 m deep, with a sand bottom and a
riffle, 0.25 m deep, with a sand and gravel bottom.
Riparian vegetation provided no cover.
Station 6 (RK 101.7)—On Skeleton Creek, 1.9 km
downstream of Station 5A. The station consisted
entirely of run habitat approximately 12 m wide
and up to 0.25 m deep. Riparian vegetation
provided no cover. The substrate was smooth, flat
rock covered with some sand or individual rocks.
Station 7 (RK 98.3)—On Skeleton Creek, 3.4 km
downstream of Station 6. The station consisted of
riffle and pool areas each having a bottom com-
posed of rocks embedded in sand. The riffle was
shallow, 0.15 m deep, and the pool was 0.45 m
deep. Riparian vegetation provided no cover.
Station 8 (RK 94.8)—On Skeleton Creek, 3.5 km
downstream of Station 7. The station consisted of a
0.3 m deep pool with a sand bottom. The riffle was
0.25 m deep and had a sand and gravel bottom.
Riparian vegetation provided no cover.
Station 9 (RK 90.6)—On Skeleton Creek, 4.2 km
downstream of Station 8. The station was com-
posed of a pool, 0.60 m deep, and a riffle, 0.30 m
deep, with a bottom of rocks embedded in sand and
clay. The creek banks were red clay and the water
was turbid. Riparian vegetation provided about 20
percent cover.
Station 10 (RK 83.2)—The most downstream
station 7.4 km farther downstream than Station 9.
The station was entirely a pool of approximately
0.76 m depth which appeared to be the result of
many wood snags creating a dam. The banks were
red clay and the water was turbid. Riparian
vegetation provided no cover.
2-2
-------
Sampling
Stations
Skeleton Creek
2
3
Fertilizer Plant
5
5A
6
7
8
9
10
1070
1048
1046
104.3
1036
101.7
98.3
94.8
906
832
Boggy Creek
1A
1
Refinery
POTW
3
28
1 2
0.7
0.5
02
Figure 2-1.
Map of study site on Skeleton Creek, Enid,
Oklahoma.
2-3
-------
3. Hydrology Survey
The purpose of the hydrology study of Boggy and
Skeleton Creeks was to obtain stream and discharge
flow measurements during the study period, and to
determine the percent flow contribution from the
three dischargers. In addition to these measure-
ments, data was also obtained from a proximate
USGS gauging station (downstream from Station 10)
and from the operational records of the dischargers.
Sampling and analytical methods are presented in
Appendix A.
3.1 Discharge Flow Measurements
Stream flows were measured 9-11 August 1 983 at
the stations shown on Table 3-1. The daily average
flows for 8-11 August for the Refinery, the Fertilizer
Plant, and the USGS gauging station (Station
07160500 near Lovell, OH) are also given. In addition,
the average POTW plant flows for 8-12 August are
presented. The Refinery reported a uniform flow of
0.023 mVsec. The daily average flow at the POTW
varied between 0.066 mVsec on 1 2 August to 0.083
mVsec on 9 August. The hourly flows at the POTW
deviated from the average values due to the facility's
day/night loading cycle. The pumps at the Fertilizer
Plant were turned on and off such that the discharge
flow was either zero or between 0.072-0.075 mVsec
(Table3-2). Durmgtheperiod8-11 Augustthepumps
were off for 7-10 hours each day. On 10 August, the
discharge was on for 22.7 hours.
Table 3-2. Effluent Pumping Records and Daily Average
Discharge at the Fertilizer Plant, Enid, Oklahoma
Date
8
9
10
1 1
Aug
Aug
Aug
Aug
Time
0000
1100
1755
2335
0000
0930
0000
0955
1115
0000
0810
- 1100
- 1755
- 2335
- 2400
- 0930
- 2400
- 0955
-1115
- 2400
- 0810
- 2030
Discharge
(mVsec)
0.072
00
0072
00
00
0075
0075
00
0072
00
0074
Daily Average
Discharge
(m3)
0050
0045
0069
0049
Source Plant operating records, personal communication
Table 3-1.
Measured Flows and Discharges on Skeleton Creek and Boggy Creek, Enid, Oklahoma
Flow(m3/sec)
Stations
8 Aug
9 Aug
10 Aug
11 Aug
12 Aug
Boggy Creek
1
Refinery3
Bb
POTW"
3
Skeleton Creek
2
4
Fertilizer Plant8
5
6
7
8
9
USGS Estimate Flows
.-
0023
--
0074
--
--
-.
0050
--
-.
--
--
-.
0259
--
0023
0043
0083
--
0045
--
0 147
--
--
0 166
0227
0.023
0081
0126
0.006
0145
0069
0 171
--
--
0148
0031
0023
0067
--
--
0049
--
0084
0 103
0131
0066
--
--
--
--
0 126
"Average flow from plant records During the night, treated wastewater was sent to the Refinery and Fertilizer Plant
"Additional stream measurement taken just downstream of the Refinery
NOTE. The confluence of Boggy Creek and Skeleton Creek is upstream of Station 4
3-1
-------
There was a large variation between the flows
measured at Stations 5 through 9 and the flows
recorded at the USGS station on 9 August (Table 3-1).
The measured flows at Station 6 (0.147 mVsec) and
Station 9 (0.166 mVsec) are much less than the
0.227 mVsec value reported by the USGS. However,
the average USGS flow decreased by 35 percent
between 9 and 10 August such that the hourly flows
on 9 August must have been decreasing continually.
Si nee the Station 6 and 9 flows were measured in the
late afternoon, they would be expected to correspond
to a lower USGS flow than the reported daily average
value. On 10 August, the Station 5 flow would be
expected to be in better agreement with the USGS
flow since the discharge from the Fertilizer Plant
stopped for only 1.3 hours. In contrast, on 11 August,
the Plant's pump had been on for 2.5 and 3 2 hours
before the flows were measured at Stations 7 and 8
which are located 6.3 and 9.8 km, respectively,
downstream from the Fertilizer Plant. It is possible
that the flow increase had not had sufficient time to
propagate downstream by the time of the measure-
ment, so that the reported value would be less than
the daily USGS values.
3.2 Flow Contribution
Using the measured flow period of 9-11 August, the
mean flows were 0023 mVsec for the refinery,
0.077 mVsec for the POTW, and 0.054 mVsec for
the Fertilizer Plant (Table 3-1). The measured up-
stream flows and the mean discharge flows sum to a
combined flow at Station 5 of 0.1 91 mVsec. The flow
of 0.191 mVsec exceeds the mean flow at the USGS
gauging station for 9-11 August of 0 1 69 mVsec by
13 percent. Assuming that water is not being lost
from the stream bed, this discrepancy could not result
from any combination of over-estimating the up-
stream flow or the reported discharges, or under-
estimating the USGS flow The higher flow of 0.1 91
mVsec was used downstream
Table 3-3. Mean Flow and Percent Flow Contribution from Three Discharges for Boggy Creek and Skeleton Creek, Enid,
Oklahoma. August 1983
Percent Flow Contribution
Station
2
1
Bb
3
4
5
6
7
8
9
Total Flow
(mVsec)
0006
0031
0054
0 131
0 137
0 191
0191
0 191
0 191
0 191
Upstream
1000
1000
574
236
270
194
194
194
194
194
Refinery
42 6
17 6
168
120
120
12 0
12 0
120
POTW
588
56 2
403
403
403
403
403
Fertilizer
Plant
283
283
283
28 3
283
bAdditional stream measurement taken just downstream of the refinery, total flow value is station 1 plus the Refinery mean flow values
Source Tables 3-1 and 3-2
3-2
-------
4. Laboratory Toxicity Tests
Toxicity tests were performed on two effluents and
water collected from nine stream stations to measure
subchronic effects on growth of larval fathead
minnows (Pimephales promelas) and chronic effects
on reproduction of Ceriodaphnia. Descriptions of the
toxicity test methods are presented in Appendix B. A
wide span of effluent concentrations were used so
acute mortality could be measured as well if it existed.
The objective of the effluent tests was to measure the
minimum concentration of each effluent that would
cause acute mortality and chronic effects on the
growth of the fat head minnows or reproduction of the
Ceriodaphnia. The ambient toxicity tests were con-
ducted to measure if toxicity exists either before or
after an effluent was discharged to estimate the
persistence of toxicity. The effect levels can then be
compared to the extant effluent concentrations in
Skeleton Creek to predict where the impact on stream
population occurs, if any. The validity of these
predictions is determined by examining the biologic
condition of the stream at the locations where the
effluent concentrations occurred as determined by
the hydrological survey.
The effluent and ambient samples were collected,
cooled, and transported to Duluth for toxicity testing.
All tests were run with one composite sample of each
effluent or stream station.
4.1 Chemical and Physical Test
Conditions
The laboratory temperature was maintained at 25 ±
1°C over the test period. Routine water chemistry
measurements for the effluent and ambient tests are
given in Table 4-1. Dissolved oxygen (DO) and pH
were monitored daily, and the initial pH, DO, con-
ductivity, hardness, and temperature measurements
were for both the fathead minnows and Ceriodaphnia
tests. ThepH values were all within 7.3 to 8.6, except
for the 100 percent Refinery effluent which was 5.7
to 6.7. The initial DO values were all 7.6 to 8.4 mg/L
except for the 100 percent Fertilizer Plant effluent
which was 6.0 mg/L. Table 4-1 also gives the mean
final DO values for the fathead minnow tests. The
Refinery effluent dilution test and the ambient station
tests had final DO values ranging from 5.1 to 6.7
mg/l. However, at the 30 and 100 percent Fertilzier
Plant concentrations the mean final DO's were 3.1
and 0.5 mg/l. The dilution water and other concen-
trations of the Fertilizer Plant had DO values ranging
from 6.2 to 4.7 mg/l. Other site studies with waters of
high BOD levels and DO levels of less than 1 mg/l
have also been encountered. In one study (Mount and
Norberg-King, in press) the average weights of the
fathead minnows were higher than the previous
studies. An assessment of this situation has led to the
conclusion that dissolved oxygen measurements
taken by the dissolved oxygen probe do not accurately
reflect the micro-environmental conditions where
the fathead minnows are living. The fathead minnows
were observed moving towards the surface of the
water where in all probability the oxygen concentra-
tions are much higher than that measured by the
dissolved oxygen probe. Apparently the behavior of
the fish causing them to stay near the surface when
the dissolved oxygen levels are low makes the test
nearly independent of low DO effects. The highest
conductivities were observed in the whole effluents.
Hardness ranged form 297 to 725 mg/L CaC03. In
Table 4-2 the final DO and pH values for the
Ceriodaphnia tests are shown. All values were within
acceptable ranges.
4.2 Effluent Toxicity Test Results
Tables 4-3 and 4-4 contain the weight and survival
data for the fathead minnow effluent tests. Survival in
the Refinery effluent was significantly lower (P <
0.05) at 30 percent while weights were significantly
lower at 10 percent. Therefore, the Acceptable
Effluent Concentration (AEC) estimate was 5.5 per-
cent (which is the geometric mean of the No
Observable Effect Concentration (NOEC) and Lowest
Observable Effect Concentration (LOEC) for the
Refinery effluent. Survival in the Fertilizer Plant
effluent was significantly lower only at the 100
percent, while the weight data was significant at the
10 percent effluent concentration. The AEC was than
5.5 percent for the Fertilizer Plant.
Table 4-5 contains the Ceriodaphnia effluent test
data as well as a quality control using laboratory
water. The mean number of young per female was
significantly lower than the dilution water young
production at 30 percent for the Refinery effluent.
This gives an AEC of 17.3 percent. The Fertilizer Plant
had a significantly lower young production compared
to the dilution water at the 30 percent effluent
4-1
-------
Table 4-1. Routine Chemistry Data for Effluent and Ambient Tests
Dissolved Oxygen
Percent Effluent
(v/v) or Ambient
Sample
Refinery
Dilution Water
(1A)
1
3
10
30
100
Fertilizer Plant
Dilution Water
(1A)
1
3
10
30
100
Ambient Station
2
3
4
5
5A
6
7
8
9
Initial
pH Range
79-80
79-80
78-80
77-80
73-80
57-67
79-80
79-80
79-8 1
7.9-80
7.9-8.0
81-82
7.7-82
78-8 1
7 9-8 2
79-81
8.1-85
8 1-8.5
79-8 2
82-83
85-8 6
Mean
Initial
(Range)
84
(82-8 9)
84
(83-87)
84
(8.3-87)
8.4
(83-87)
83
(78-87)
76
(67-84)
82
(77-86)
82
(74-86)
82
(76-8 7)
8.3
(76-89)
82
(79-87)
60
79
(73-84)
79
(68-8 5)
7 6
(62-84)
77
(60-85)
82
(70-88)
82
(7.6-86)
80
(62-86)
85
(82-88)
85
(80-88)
Mean
Final"
(Range)
60
(46-6 9)
64
(47-71)
6 1
(50-71)
6.4
(54-70)
64
(5.3-74)
67
(64-70)
62
(53-72)
60
(5 6-6 5)
55
(46-60)
47
(37-5 6)
3 1
(26-42)
05
(-)
70
(59-82)
60
(50-80)
60
(4.5-7 6)
5 1
(3 1-6 8)
60
(43-7 8)
6 1
(44-7 6)
54
(39-69)
62
(49-7 6)
66
(55-81)
Temperature
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
25
Hardness
(mg/L)
297
__
-.
-_
297
-.
__
-_
-.
--
477
376
380
725
466
663
730
700
710
Conductivity
(/umhos/cm)
1,220
1,280
1,340
1,580
2,320
4,680
1,280
1,320
1,410
1,800
2,700
5,900
1,620
1,980
2,040
3,480
2,050
2,880
3,500
3,390
3,200
Final dissolved oxygen values are for fathead minnows tests only Table 4-2 contains Cenodaphnia final chemistry values
concentration, which gives an AEC of 1 7 3 percent
The quality control sample young production was in
the normal range (Mount and Norberg, 1984).
4.3 Ambient Toxicity Test Results
Tables 4-6 and 4-7 contain the survival and weight
data for the fathead minnow ambient tests Station 5
had significantly lower survival (P < 0.05), while the
weights of Stations 3, 4, 5, 5A, 6, and 8 were
significantly lower when compared to Station 9. Table
4-8 presents the results of the survival and mean
young production of the Cenodaphnia. Stations 2, 5,
7, and 8 had significantly lower mean number of
young per female ismg th highest value of young
4-2
-------
Table 4-2. Final Dissolved Oxygen and Final pH for
Ceriodaphnia Effluent and Ambient Toxicity
Tests
Percent Effluent
or Ambient
Sample
Refinery
Dilution Water (1 A)
1
3
10
30
100
Fertilizer Plant
Dilution Water (1 A)
1
3
10
30
100
Ambient Station
2
3
4
5
5A
6
7
8
9
pH
8 1
82
8.4
8.4
83
7 1
74
8.3-84
84
8.3-8.4
82-83
7.9
85
8.4
84
85
84
84
8.3
8.4
8.4
Mean DO
(mg/L)
78
7.8
79
78
78
79
79
7.5
78
72
68
48
7.9
79
79
79
78
78
79
8.0
8.0
DO Range
73-82
74-82
74-84
74-83
7 5-83
76-82
77-8.3
7.2-78
72-8.1
6.9-74
63-7.1
4.2-62
72-82
72-84
72-8.4
7.3-83
7.3-83
73-83
74-84
77-83
77-83
production (Station 9) for the comparison. Survival
was significantly lower only at Station 3.
4.4 Discussion
For the effluent dilution tests the fathead minnows
were affected at concentrations lower than the
Ceriodaphnia. The upstream water (1 A) used as the
dilution water resulted in good growth of the fathead
minnows and high young production of the Cerio-
daphnia. Station 9, which is the most downstream
station, produced the best growth for the fathead
minnows and the highest young production for the
Ceriodaphnia. It appears that all effects of toxicity
were removed at the downstream location.
Table 4-3. Seven-Day Survival of Larval Fathead Minnows in Two Effluents
Percent Effluent (v/v)
Effluent
Refinery
Fertilizer Plant
Replicate
A
B
C
D
Mean
A
B
C
D
Mean
100
0
0
0
0
Oa
0
0
0
0
Oa
30
30
30
50
60
43"
80
80
80
70
78
10
80
90
90
100
90
80
90
90
80
85
3
90
90
90
100
93
80
90
100
90
90
1
80
100
90
100
93
90
100
100
100
98
Dilution
Water (1 A)
100
90
90
90
93
90
100
80
90
90
"Significantly lower from the dilution water (P < 0 05)
4-3
-------
Table 4-4. Mean Individual Dry Weight (mg) After Seven Days for Larval Fathead Minnows Exposed to Two Effluents
Percent Effluent (v/v)
Effluent Replicate
Refinery A
B
C
D
Weighted Mean"
SE
Fertilizer Plant A
B
C
D
Weighted Mean"
SE
100
0
0
0
0
Oa
--
0
0
0
0
0"
30
0 19
024
032
030
0276a
0038
0.33
024
031
0.31
0297a
0.030
10
038
040
041
047
0418a
0026
0.46
051
0.46
0.56
0497°
0029
3
056
061
052
056
0562
0026
057
057
0.66
0.59
0.600
0028
1
057
058
061
063
0599
0026
054
058
065
058
0589
0.027
Dilution
Water (1 A)
056
064
052
061
0608
0018
058
0.57
065
075
0.608
0019
"Significantly lower from the dilution water (P > 0 05)
"Explanation of weighted mean calculation is in Appendix B
Table 4-5. Percent Survival and Young Production of
Ceriodaphnia in Two Effluents
Sample
Refinery
DilutionWaterfl A)
1
3
10
30
100
Fertilizer Plant
DilutionWater(IA)
1
3
10
30
100
Lake Superior Water"
Mean
Percent
Survival
100
100
100
100
100
Oa
100
90
100
90
70
Oa
90
Mean
Number of
Young per
Female
335
348
31 5
374
23 5a
Oa
253
25 1
250
31 5
1 1 4a
Oa
181
Confidence
Intervals
275-395
292-404
268-362
33 2-41 7
185-285
--
189-31 7
193-31 0
193-307
246-385
78-151
--
14 1-22 2
"Significantly different from the dilution water for each test (P <
005)
"Quality Control water sample
Table 4-6. Seven-Day Percent Survival of Larval Fathead
Minnows in the Ambient Toxicity Test
Station Number
Replicate
A
B
C
D
Mean
2
100
90
100
100
98
3
100
100
90
80
93
4
80
90
80
80
83
5
10
60
50
60
45a
5A
90
90
100
100
95
6
90
90
90
80
88
7
100
80
80
80
85
8
100
90
100
90
95
9
90
90
90
70
85
Significantly lower from Station 9 (P < 0 05)
4-4
-------
Table 4-7. Mean Individual Dry Weight (mg) After Seven Days for Larval Fathead Minnows in the Ambient Toxicity Test
Station Number
Replicate
A
B
C
D
Weighted Mean"
SD
2
066
069
063
061
0646
0033
3
049
043
058
048
0494a
0034
4
046
059
052
056
0525a
0036
5
058
031
028
042
0429a
0040
5A
063
060
069
075
0670a
0033
6
063
066
0 59
063
0627a
0035
7
069
076
069
078
0728
0035
8
056
068
065
072
0650a
0033
9
071
082
082
068
0762
0035
8Significantly lower from Station 9 (P < 0 05)
"Explanation of weighted mean calculation is in Appendix B
Table 4-8. Percent Survival and Mean Young Production of
Ceriodaphnia in the Ambient Toxicity Test
Ambient
Station
2
3
4
5
5A
6
7
8
9
Percent
Survival
100
30a
90
70
80
100
80
60
80
Mean
Number of
Young per
Female
11 7a
140
164
81a
228
190
128a
99a
243
Confidence
Intervals
99-135
106-173
11 5-21 3
58-103
187-268
160-220
100-155
58-14 1
179-304
"Significantly different from Station 9 (P < 0 05)
4-5
-------
5. Plankton Community Survey
This survey investigated the plankton community by
measuring the occurrence and density of organisms
in Skeleton Creek and Boggy Creek. The primary
emphasis was to collect zooplankton, but algae were
also collected and enumerated. The number of
species and individuals are used to determine altera-
tions in composition and/or density. The sampling
and analytical methods are presented in Appendix C
Samples were not collected at Stations 1 and 10.
5.1 Community Structure
Algae were the dominant planktonic organisms at
every station on Skeleton Creek and Boggy Creek
(Table 5-1). The number of algae were lowest at
Stations 1A and 2, but the numbers increased at
Stations 3 and4 by 8 and 7.5 times, respectively. The
highest algal densities were found at Stations 5, 6,
and 8 where there were over 8,000 organisms/liter.
Solitary diatoms composed the great majority of the
algal population.
The numbers of crustaceans and rotifers collected
were low. Often the only crustaceans found were
nauplii. The highest density of crustaceans was 2.44
organisms/liter at Station 7 At Station 5, no crusta-
ceans were collected. The highest density of rotifers
was also at Section 7 (30.65 organisms/liter). Non-
loricate forms composed the majority of the rotifers
and the remainder were from the Family Branchi-
onidae.
5.2 Evaluation of the Plankton
Community
Rotifers were the most abundant zooplankton group
in Skeleton Creek and Boggy Creek. In general,
densities of rotifers were low at the upstream
stations, but consistently increased downstream to a
Table 5-1. Mean Density (number/liter) of Planktonic Organisms Collected in Skeleton Creek and Boggy Creek, Enid, Oklahoma,
August 1983
Sampling Station
Taxa
Crustaceans
Cladocera
Copepoda
Naupln
Total crustaceans
Rotifers
Branchionus spp.
Small Branchionidae
Non-loricate forms
Total rotifers
Algae
Padiastrum
Desmids
Solitary diatoms
Total algae
Others
Chironomidae
Tnchoptera
Heleidae
Nematoda
Total others
Total Zooplankton Taxa
1A
0 15
0.15
004
0 14
--
018
005
072
112 1
11287
019
--
004
005
028
3
2
--
.-
024
024
--
048
--
048
0 12
1 67
1490
15079
024
--
--
0 12
036
2
3
056
006
1 45
207
074
1 88
251
513
063
048
9082
90931
048
--
--
0 12
0.60
5
4
007
006
031
044
026
071
4 21
5 18
068
063
1,1165
1,11781
032
--
--
0.32
5
5
--
--
099
1 21
5 61
781
1 21
062
9,435 9
9,437 73
022
--
022
044
3
5A
--
--
005
005
1 33
2 12
7 12
1057
1 14
002
2,382 5
2,383 66
1 36
007
1 43
4
6
034
034
326
200
933
1459
7 16
011
2,2383
2,245 57
085
--
--
028
1 13
4
7
026
2 18
2 44
7 22
5 87
17 56
3065
7 28
049
5,573 6
5,581 37
061
--
039
1.00
4
8
--
055
055
0 55
1 54
1 65
3 74
6 50
--
12,7275
12,7340
033
--
022
055
4
9
004
004
070
088
082
240
728
096
3,309 6
3,317 84
004
004
008
4
Note. -- indicates organisms were not found.
5-1
-------
30 i
o
o
N
'o
OJ
a)
_Q
E
Crustaceans
Rotifers
a Algae
J2 20-
10-
r 12,000
-10,000
-8,000 z
c
3
cr
\
6,000 5"
- 4,000 £•
- 2,000
Figure 5-1.
1A
Sampling Stations
Densities of crustaceans, rotifers, and algae at Skeleton Creek and Boggy Creek, Enid, Oklahoma, August 1983.
maximum at Station 7, and densities decreased
below Station 7 (Figure 5-1). Results of a one-way
analysis of variance (ANOVA) indicated that the
difference in densities between stations was highly
significant(P = 0.0001) Tukey'stesUSokal and Rohlf,
1981) results indicated that Station 6 and 7 were
significantly different (P < 0.05) from all other
stations.
Despite the low densities of crustaceans, there were
significant differences (P = 0.0001) between stations
from results of a one-way AIMOVA Tukey's test
results indicated that Stations 3 and 7 were signif-
icantly different (P < 0.05) from the other stations
Crustacean densities were highest at Station 3 and 7
while less than 0.6 organisms/L at other stations
(Figure 5-1)
5-2
-------
6. Macroinvertebrate Community Survey
The survey investigated the macroinvertebrate com-
munity of Boggy Creek and Skeleton Creek. Samples
were collected from natural and artificial substrates.
The macroinvertebrate community is considered to
be a good indicator of changes in water quality due to
their limited mobility. The degree of community
stability can be ascertained by measuring species
composition and dominance. An alteration in com-
munity structure, species composition, or biomass
beyond normal variations would be regarded as an
adverse effect. In addition, the increased abundance
of nuisance insect larvae or other benthic species
would be regarded as an adverse effect.
Although both natural and artifical substrates were
used to quantify the macroinvertebrate communities,
not all stations were sampled by both methods (Table
2-1 and Appendix C). A description of the sampling
and analytical methods is presented in Appendix C.
Additional data are included in Appendix D.
6.1 Community Composition
The macroinvertebrate communities of Boggy Creek
and Skeleton Creek were composed of 55 taxa. The
number of taxa at each station varied from 1 3 to 28.
Major taxa were identified as those which contributed
a minimum of 5 percent of the total number of
organisms from at least one station. The changes in
abundance and percent composition of these major
taxa are presented for the two substrate types.
6.1.1 Natural Substrates
Two taxa were more abundant than the other major
taxa. a chironomid—Dicrotendipes sp. and a cole-
opteran—Berosus sp. Dicrotendipes composed over
30 percent of the benthic density at Stations 2, 3, and
5, and Berosus constituted over 30 percent of the
benthos at Stations 5A, 6, and 7 (Table 6-1). There are
another fifteen taxa which contributed >5 percent of
the populations for at least one station. These taxa
were in six taxonomic groups: Diptera (Chirono-
midae), Ephemeroptera, Odonata, Trichoptera, Gas-
tropoda, and Oligochaeta.
The macroinvertebrate population in Skeleton Creek
and Boggy Creek is primrily composed of insects.
Fifteen of the seventeen major taxa are insects. The
Chironomidae family (midges) had the most taxa (9).
Two taxa each were from the Ephemeroptera (may-
flies) and Trichoptera (caddisflies) families. One
major taxon each were identified from the Coleoptera
(beetles), Odonata (dragonflies, damselfhes), and
Physidae (pouch snails) families, and Oligochaeta.
6.1.2 Artifical Substrates
The same two taxa were found to be most abundant
using the artifical substrates as with the natural
substrates: a chironomid—Dicrotendipes sp. and a
coleopteran—Berosus sp. (Table 6-2). Dicrotendipes
sp. composed greater than 30 percent of the macro-
invertebrates population at five stations. In contrast,
Berosus sp. composed approximately 50 percent at
Station 6, 27 percent at Station 7, and less than 6
percent at the other stations. There are another
fourteen major taxa which contnbuted>5 percent of
the populations for at least one station. These taxa
were insixtaxonomicgroups: Diptera(Chironomidae),
Ephemeroptera, Odonata, Trichoptera, Amphipoda,
and Gastropoda.
The macroinvertebrate population, collected using
artificial substrates, in Skeleton and Boggy Creek is
primarily composed of insects. Fourteen of the sixteen
major taxa are insects. The Chironomidae family
(midges) had the greatest number of major taxa
(eight). Similar to the results for natural substrates,
two taxa each were from the Ephemeroptera (may-
flies) and Trichoptera (caddisflies) families and one
each from the Coleoptera (beetles), Odonasta (drag-
onflies, damselflies), Physidae (pouch snails), and
Tahtridae (scuds) families.
6.1.3 Comparison Between Substrate Types
The taxa collected from the natural and artificial
substrates for dominant taxa were very similar The
most abundant major taxa were the same for the two
substrates. A difference between the two substrates
occurred m the non-insect taxa Physidae and Oligo-
chaeta were the non-insect major taxa collected from
the natural substrates while Physidae and Taltridae
were the non-insect major taxa collected from the
artifical substrates.
6-1
-------
Table 6-1. Mean Percent Composition of Major Macroinvertebrate Taxa Collected from Natural Substrates in Skeleton Creek
and Boggy Creek, Enid, Oklahoma, August 1983
Taxa
Diptera
Dicrotendipes sp
Polypedilum sp
Ab/abesmyia sp
Chironomus sp
Tanypus sp
Tanytarsus sp
Pseudochironomus sp
Cr/cotopus sp
Chironomidae pupae
Total Chpronomidae
Ephemeroptera
Caenis sp
Baetis sp
Coleoptera
Berosus sp
Odonata
Argia sp
Trichoptera
Cheumatopsyche sp
Hydropsyche sp
Gastropoda
64
64
1
39
Sampling Station
1A
250
109
45
1 0
03
64
05
1 0
2.3
545
178
08
2
400
58
05
09
30
137
58
726
09
3
349
1 3
08
365
24
78
05
46
89 5
1
4
183
05
26
170
200
12 8
-.
08
731
5
365
09
58
164
05
1 2
10 1
189
909
07
5A
16 1
04
2 9
92
1 3
1 2
--
46
79
43 6
1
--
6
145
07
3 7
02
05
02
11 2
68
38 7
05
--
7
108
02
1 1 7
05
02
1 9
02
15 9
4 1
45 5
1 8
1 6
8
5 1
31 2
82
--
16 8
05
82
8 1
78 7
05
9
03
185
1 0
»
1 4
02
1 8
23 2
7 6
08
152
29
01
30 1
01
400
1 3
02
428
1 3
Source Table D-1
Note -- indicates not collected
63
44
03
Physidae
Oligochaeta (unidentified)
21
33
67
125
03
7 8
69
05
23
08
03
21 1
2 2
79
58
7 1
09 C
90 e
1 2
43 7
105
03
6 1
6.2 Station Comparisons
6.2.1 Natural Substrates
The greatest number of organisms collected from
natural substrates was at Station 5 and Chironomidae
taxa comprised 90 percent of these (Tables D-1 and
6-1) Collections were greater than 2,100 organ-
isms/m2, except at Stations 1A, 2, and 3 where
collections were less than 1,500 organisms/m2
There are noticeable differences in the abundance of
many of the major taxa between stations. There were
also differences in the abundance patterns between
taxa. The mean density of Dicrotendipes sp. varied by
over two orders of magnitude, from a maximum at
Station 5 of 1,568/m2to only 7/m2 at Station 9
(Figure 6-1). Mean densities of Dicrotendipes sp. at
the other stations were 120-620/m2. Results of a
two-way ANOVA indicated that these differences in
numbers between stations were highly significant (P
= 0.001), and results of Tukey's test indicated that
Station 9 was significantly different (P <0.05) than all
the other stations, except Static. \ G. ~\\ ie mean density
of Berosus sp. varied by two orders of magnitude from
a low of 11 /m2 at Station 3 to a maximum at Station
5A of 1,159/m2 (Table D-1, Figure 6-2). Highly
significant differences (P = 0.0001) were found in
numbers of Berosus sp uetwee "> .-Uions from an
ANOVA and Tukey's test results indicated that Station
3 was different (PS 0 05) than Stations 6 and? where
Berosus composed at least 40 percent of the com-
munity
Examination of the abundance trends for the two
most abundant macroinvertebrate taxa collected from
natural substrates indicated that densities of Dicro-
tendipes sp. peaked distinctly at Station 5, while
densities of Berosus sp. peaked immediately down-
stream at Station 5A (Figures 6-1 and 6-2). Other
major taxa also had maximum densities at Stations 5
or 5A: Chironomus sp., Cricotopussp., Chironomidae
pupae, and unidentified oligochaetes. The contribu-
tion of the Chironomidae to the composition at each
station was overwhelming at Stations 3 and 5, where
that family composed 90 percent of the taxa. In
contrast, Polypedilum sp., was found in greatest
abundance at Stations 8 and 9 where other chirono-
mids were least abundant. Cheumatopsyche sp. was
also most aounaant at Station 9.
There were nonsignificant differences between sta-
tions of the number of chironomid taxa However, for
the total number of taxa, ANOVA results indicated
that there were very signficant differences (P - 0 007)
6-2
-------
Table 6-2. Mean Percent Composition of Major Macroinvertebrate Taxa Collected from Artificial Substrates in Skeleton Creek
and Boggy Creek, Enid, Oklahoma, August 1983
Taxa
Diptera
Dicrotendipes sp
Polypedi/um sp
Ablabesmyia sp
Chironomus sp
Psectrocladius sp
Tanypus sp
Tanytarsus sp
Chironomidae pupae
Total Chironomidae
Ephemeroptera
Caems sp
Baetis sp
Coleoptera
Berosus sp
Odonata
Argia sp
Trichoptera
Cheumatopsyche sp
Hydropsyche sp
Amphipoda
Tahtridae
Gastropoda
Physidae
1
580
06
14 1
5 5
07
46
44
89 5
70
04
--
2 1
09
2
63 7
07
67
04
1 5
24
95
86 7
1 9
1 3
20
63
--
6 1
3
450
07
330
36
5 3
^0 1
<0 1
44
942
02
03
02
30
07
02
4
53 3
02
13 1
1 0
2 4
2 2
<0 1
2 2
762
08
1 9
62
5 5
6 6
12 7
Samplmi
5
31 3
03
15 1
146
05
105
0 1
11 7
935
03
1 5
3 1
05
--
07
03
3 Station
6
7 2
02
8 6
54
1 4
1 4
05
25
288
04
8 5
52 3
5 4
02
1 2
09
7
260
0 1
4 1
103
2 9
2 5
04
09
483
1 0
160
270
4 5
2 5
0 2
8
06
402
4 9
0 1
08
21 8
5 5
74 1
1 3
4 3
08
13 6
3 7
0 1
02
9
2 9
374
43
-.0 1
08
^01
48
3 1
55 6
2 7
2 8
3 1
31 1
10
3 8
25 3
192
04
1 0
14 7
71 6
04
17 5
94
Source Table D-2
Note -- indicates not collected
2,400-
, Artificial Substrates
Natural Substrates
Artificial Substrates
Natural Substrates
1A
Figure 6-1.
3 4 5 5A 6 7
Sampling Station
Mean densities of Dicrotendipes sp. collected
from Skeleton Creek and Boggy Creek, Enid,
Oklahoma. August 1983.
3 4 5 5A 6
Sampling Station
7 8 9 10
Figure 6-2.
Mean densities of Berosus sp. collected from
Skeleton Creek and Boggy Creek, Enid, Okla-
homa, August 1983.
6-3
-------
between stations and Tukey's test did indicate that
Stations 1A and 5 were significantly different (P ^
0 05) from Station 3 The lowest number of taxa were
found at Stations 2, 3, and 8 with 13-14 taxa (Table
D-1). The greatest number of taxa occurred at Stations
1 A. 5, and 6 with 24-25 taxa
5.2.2 Artifical Substrates
The greatest number of organisms collected by the
artificial substrates was at Station 8 and of these 75
percent were from the Chironomidae family (Tables
D-2 and 6-2) The mean number of organisms
collected at Station 8 (14,951) is a I most two and one-
half times greater than at next highest values at
Stations 3 and 7.
Similar to the data collected from natural substrates,
there are differences in abundance between stations
and in patterns of abundance for the major taxa For
one of the two most abundant major taxa, Di-
crotendipes sp., peak mean densities were observed
at Stations 3 and 7 with variations of up to two orders
of magnitude (Figure 6-1). These station differences
were highly significant (P = 00006) as shown by
ANOVA results on the number of Dicrotendipes sp
and the Tukey's test results indicated that Station 3
was significantly different (P <0 05) than Stations 8,
9, and 10. Mean densities of Berosus sp varied by
approximately two orders of magnitude with a
maximum density at Station 7 of 1,755 organisms/m2
(Figure 6-2). Results of an ANOVA indicated that,
similar to the case with natural substrates, the
differences in Berosus sp. abundance between
stations was highly significant (P = 00001). In
addition, Stations 6, 7, and 8 were different (P <
0.05), due to the •'• jh abundance from all other
stations. Peak mean densities also occurred at Station
7 for Baetis sp (Ephemeroptera) and Argia sp
(Odonata), and at Station 8 for Polypedilum sp and
Janytarsus sp. (Chironomidae)
Examination of thetotal number of macroinvertebrate
taxa by ANOVA indicated that there were highly
significant differences (P = 0.0001) between stations.
Tukey's test results indicated that Station 8 was
significantly different (P < 0.05) from Station 10. The
number of chironomid taxa varied between stations,
and these differences were significant (P - 0.045)
according to ANOVA results, and Tukey's test results
did not show significant differences. This finding is
consistent with the results from the natural sub-
strates—that there were no differences in the number
of chironomid taxa between stations.
6.2.3 Gear Comparison
While the same major taxa were collected by collect-
ing gears, the numbers and locations of taxa were not
the same. In addition, the total number of organisms
collected was generally higher using the artificial
substrates (Table 6-3) However, these differences
are expected due to the nature of the two substrates
The artificial substrates are composed of smooth,
homogenous surfaces which were suspended in the
water column for 20 days and are relatively immobile
during that period. The natural substrate of Skeleton
Creek is principally bedrock with overlying shifting
sand The shifting sand offers some degree of
instability to benthic fauna
The Chironomidae family was the most abundant of
any of the macroinvertebrate groups collected using
either the natural or the artificial substrates. The
number of chironomid taxa was similar for all stations
for both substrates. The Chironomidae (midges)
composed the largest proportion of their community,
up to 90 percent at Station 5, with few exceptions.
From the natural substrates, a trichopteran (caddisfly)
composed over 40 percent of the community at
Station 9 and from the artificial substrates, a cole-
opteran (beetle) composed over 50 percent of the
community at Station 6.
There were two taxa which were noticeably more
abundant in Skeleton and Boggy Creek using either
substrate. The most abundant of the major taxa,
Dicrotendipes sp. (a chironomid) showed highly
significant differences in numbers between stations
for both substrates The second most abundant taxon,
Berosus sp. (a coleopteran) also showed highly
significant differences in numbers between stations
for both substrates
The number of chironomid taxa did not change
significantly between stations However, the abun-
dance and composition forthe other major taxa varied
between the two types of substrates, and the total
number of taxa did vary significantly between sta-
tions. For the natural substrates, the number of taxa
collected at Station 3 was significantly lower than the
number collected at Stations 1A and 5 The number of
taxa collected by artificial substrates at Station 8 was
significantly greater than Station 10
6.3 Evaluation of the Macroinvertebrate
Community
The macroinvertebrate community of Skeleton Creek
and Boggy Creek was dominated by insects. The most
abundant taxonomic group was the Chironomidae
family (midges). Other major taxa were from the
Coleoptera (beetles), Ephemeroptrae (mayflies), Tri-
choptera (caddisflies), and Odonata (dragonflies,
damselflies) families.
The community composition changed between sta-
tions. Statistically signficant differences between
stations were found using the total number of taxa
collected; although the stations which were different
6-4
-------
Table 6-3. Total Taxa Collected by Artificial and Natural Substrates and the Combined Macroin vertebrate Taxa at Each Sampling
Station
Sampling Station
Natural
Substrate
Artificial
Substrate
Combined
Substrate
Total"
1A 1 2
25 -- 14
13 21
25
3
13
20
22
4
16
21
25
5
23
20
29
5A 6
16 24
23
32
7
18
25
28
8
13
28
30
9 10
19
18 13
25
"Total number of unique taxa in either natural or artificial substrate sample, numbers were tallied using Tables D-1 and D-2
NOTE -- Means no sample was available, see Chapter 2 for clarification
varied with substrate type In addition, changes in the
number of taxa for the most abundant group, the
chironomids, were nonsignificant between stations
6-5
-------
7. Fish Community Survey
This study investigated the fish community in Skel-
eton Creek and Boggy Creek Species abundance and
composition were used as measures of community
stability A description of the sampling and analytical
methods is in Appendix C. A list of fish species and
families are given in Table D-7
7,1 Community Structure
The fish community at Skeleton and Boggy Creek was
composed of 1 1 taxa (Table 7-1) These taxa repre
sented five families of fish Three species were
present at all but one station red shiner (Notropis
lutrensis), sand shiner (Notropis stramineus), and
mosquitofish (Gambusia affmis) although the latter
was not abundant The most abundant taxa were the
red and sand shiners, and the early juveni lecypr in ids
(minnows). Most of the taxa collected in Skeleton
Creek and Boggy Creek were from the Cyprmidae
(minnows) or Centrarchidae (sunfish) families
The number of fish taxa collected varied between 3-7
per station except at Station 5 where none were
caught. The largest number of fish taxa were col-
lected at Stations 2, 3, and 9 and the least at Station 7
and 8 The greatest numbers of fish were caught at
Stations 3 and 6. The proportion of run habitat at
Station 3 was 50 percent and approximately 100
percent at Station 6 At Station 3, red shmers
composed over 35 percent of the catch (Table 7-1) In
contrast, at Station 6, red shiners composed over 75
percent and sand shmers composed over 20 percent
of the catch Total number of other fish species
caught were quite low, under 20 fish per station, with
the excention of the mosquitofish
7.2 Evaluation of the Fish Community
Another fish survey of Boggy and Skeleton Creek had
been conducted in 1982 (JRB Associates, 1983) In
that survey, four stream collection stations were used
in locations similar to Stations 1,3,5, and 9 used in
this study The number of fish taxa varied between 2.
and 6 m that earlier study (JRB Associates, 1 983) Of
the six species they collected, the most abundant was
the red shiner Lower catches and numbers of taxa at
the two intermediate sites were regarded by JRB
Associates (1 983) as indicative of degradation
Results of this 1 983 Skeleton Creek survey revealed
higher catches of fish and more numbers of taxa than
previously (JRB Associates 1983) Results of X*1 test
on the number of taxa indicated no significant
difference between stations, using either Station 1 A
(upstream) or Station 2 (maximum) as the expected
value
Table 7-1 Number of Fish Collected by Seine from Skeleton and Boggy Creeks, Enid, Oklahoma, August 1 983
Taxa
1A
Sampling Station
55A
Notropis lutrensis
Notropsis stramineus
Notropis umbratilis
Pimephales promelas
Phenacobis mirabihs
Notemigonus crysoleucas
Lepomis megalotis
Lepomis humilus
Lepomis cyanellus
Ictalurus melas
*? TO b U S ! 3 n**--'~
Early juvenile Notropis
Early juvenile Carpoides
Total number of fish
Total number of taxaa
79
24
3
3
--
--
398
507
4
18
7
1
5
5
--
5
24
71
136
7
500
2
20
2
4
1
800
1,329
6
47
874
1
1
R
30
959 0
5 0
15
22
1
3
1
42
4
83-5
232
12
1 1
1
1,091
4
98
55
53
2
208
3
176 294
349 26
19
3
1
27 1
25
552 369
3 6
"X2 test results indicate nonsignificant differences between stations using either Station 1 A or Station 2 as the expected value
7-1
-------
The number of species is similar between stations
which indicates that the community structure is
unchanged between stations However, there were
fluctuations in the number of fish collected Most
notable was the paucity of the fish collected at Station
5A, where very few of the abundant red shiner, sand
shiner, and young juvenile cyprmids were caught
7-2
-------
8. Comparison Between Laboratory Toxicity Tests and Instream Biological Response
8.0 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.
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 mean 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. Therefore, a lower number of taxa should be
a predictable response of the community For ex-
ample, there should be a relationship between the
number of young per female Ceriodaphnia or the
growth of fathead minnows (or other test species) and
the number of species in the community. Obviously,
the test species must have a sensitivity, such that at
ambient concentrations to which the community has
responded, a partial effect is produced in the toxicity
test. However, unless the special case described
above exists, the correlation between toxicity and
species richness will not be a tight one
Effluents differ from single chemicals in some
important respects We know from the literature on
single chemicals that there usually are large dif-
ferences 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
Ceriodaphnia 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 nearthedischarge point' Therefore, it is logical
to expect that sometimes one test species would be
more sensitive to the effluent as it is discharged and
another species more sensitive after fate processes
begin altering the effluent To be sure, the source of
the effluent is the same but it is certainly not the same
"effluent" in regard to its composition. If these
statements are true then one should also expect that
species in the community in the receiving water will
8-1
-------
be affected at one place near the discharge and a
different group of species will 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 by Mount
et al., 1 984 and in the studies soon to be published
(i.e., Mount eta I., 1 985), it is clear that there is no one
community component that is consistently sensitive.
Sometimes the benthic invertebrates and the peri-
phyton have similar responses and both are different
from the fish. Sometimes the fish and periphyton
have similar responses and these are unlike the
benthic invertebrates.
The same is true of the test species. Sometimes the
Ceriodaphnia respond like the periphyton and other
times like the fish community. 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 or zocplankton,
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 represent-
ative of some components of the community. The
important distinction is that one never can be sure
which components they will represent.
In comparing toxicity test results to community
response, comparison must be made with the above
in mind. Certainlythose community components that
are most sensitive will be most impacted and/or lost.
The response of the most sensitive test species
should therefore be used to compare to the response
of the most sensitive of the community.
A weakness in using the number of species as the
measure of community response is that species may
be severely affected yet not be absent. The density of
various species is greatly influenced by competition
for available habitat, predation, grazing, and/or
secondary effects which may result from changing
species composition Density is more subject to
confounding causes, other than direct toxicity, and is
not as useful as the species richness in the com-
munity to compare community response to measured
toxicity.
Several measures of community structure are based
on number of species, e g , diversity and community
loss index. Since diversity measures are little affected
by changes in the number of species (or taxa) that are
in very low densities in the community, diversity is an
insensitive measure for some perturbations which
can be measured by toxicity tests. The community
loss index is based only on the presence or absence of
specific species relative to a reference station and
would be useful except that habitat differences
between stations heavily affect this measure There
are several problems when using the number of (taxa)
species measured The foremost is that the mere
presence or absence of species is not a compre-
hensive mdictor 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 dueto either drift, immigration, or
some catastrophic event when, in fact, that species is
not actually a part of the community where it is found.
Effects other than toxicity, such as habitat, will
always confuse such comparisons to toxicity data to
some extent. They cannot be eliminated Identifica-
tion of taxa to different levels can reduce the
sensitivity of species richness. Even though species
richness has numerous sources of error as a repre-
sentative measure of community health, it remains
the best measure for comparison with toxicological
data. Species sensitivity will respond in the most
direct way to toxic response of the community with
the least interference.
8.1 Prediction of Instream Community
Impacts Based on Effluent Dilution Test
Results
The calculated Acceptable Effluent Concentration's
(AEC) for each test species and effluent tested are
presented in Table 8-1, as well as the Instream Waste
Concentration (IWC) for each effluent downstream of
the discharge The AEC is based on the most sensitive
endpoint of the most sensitive species. The Refinery
IWC was about three times higher than the AEC while
the IWC of the Fertilizer Plant was about five times the
AEC. Based on these results, there should be
noticeable ambient toxicity at the stations below each
discharge and adverse effects on the mstream bio-
logical community since some species would be
8-2
-------
Table 8-1
Comparison Between the Acceptable Effluent Concentration (AEC) and the Instream Waste Concentration JIWC) for
Effluents Tested
AEC (Percent)
IWCC (Percent)
Effluent
Refinery
Fertilizer Plant
Fathead
Minnow"
5 5
5 5
Cenodaphnia3
17 3
17 3
Station 3
176
Station 5
120
283
Calculated from data in Table 4-4
"Calculated from data in Table 4-5
GData from Table 3-3
expected to be as sensitive as the most sensitive test
species.
For Station 3 below the Refinery, the IWC was
estimated at 17.6 percent, which was much higher
than the AEC. Therefore, toxicity instream at Station
3 was predicted, and ambient toxicity was increased
at Station 3 (Table 8-2) At Station 4, Skeleton Creek
and Boggy Creek have joined and a slight decrease in
ambient toxicity was expected and was observed.
Si nee the IWC of the Fertilizer Plant was five times the
AEC at Station 5, ambient toxicity was predicted. The
results of the ambient toxicity tests at Station 5
corroborated the prediction of the effluent dilution
test by showing increased toxicity at Station 5. The
prediction of impact at Station 5 could also have been
made using the IWC of the Refinery (Table 8-1)
8.2 Comparison of Ambient Toxicity Test
Results and Field Data
In order to make a prediction of impact from single
species data, the station with the least toxicity or the
most numbers of taxa was considered the least
impacted and used as zero percent 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 reduced species richness) could have
used a different reference station as zero percent
impact (Table 8-2). The data for the number of benthic
macroinvertebrate taxa from both the artificial and
natural substrates were combined in order to obtain a
total number of taxa found at each station where both
kinds of samples were collected The comparisons on
Table 8-2 include Stations 2 through 9 only as Station
1A was sampled for macromvertebrates on natural
substrates only since Station 1 had become im-
pounded by a beaver dam after the artificial substrates
were set. This made the comparisons of the natural
and artificial substrates impossible as the locations
and the conditions the invertebrates were exposed to
were quite different. Also, since Station 5A was
added during the August field sampling no artificial
substrate sample was collected and therefore 5A is
eliminated from the overall comparison too The
zooplankton data are of limited value as few crus-
taceans and rotifers were collected. The trends of the
percent increase in toxicity as predicted by combining
the ambient toxicity test data are compared to the
percent reduction in the number of taxa for the
various biological field components in Table 8-2
Table 8-2. Percent Increase in Toxicity and Percent Reduction in Number of Taxa for the Instream Biological Community"
Station
2
3
4
5
6
7
8
9
Cenodaphnia
Young
Production
52
42
32
66
22
47
59
0
Fathead
Minnow
Weight
15
35
31
44
18
4
15
0
Zooplankton
Taxa
60
0
0
40
20
20
20
20
Combined
Macroinvertebrate
Taxa"
22
31
22
9
0
12
6
22
Fish
Taxa
0
14
29
100
43
57
57
14
aPercent values were obtained by using the highest value for each measurement as the basis for zero percent impact
"This is the total number of unique taxa found in either the artificial or natural substrates and totalled for the comparison, see Table 6-3
Sources Tables 4-6, 4-7, 5-1, 6-3, and 7-1
8-3
-------
Table 8-3. Percent of Correct Predictions Using Four Levels of Defined Impact
Combined
Toxicity
Data (Percent)
20-100
40-100
60-1 00
80-100
20-100
875
62 5
125
0
Combined Biological t-it
40-100
750
750
500
375
;ld Data Percent
60-100
375
67 5
100
750
80-100
250
500
100
87 5
Source Table 8-3
Table 8-3 was constructed in the following manner. If
both the toxicity data and all biological field data
values were below 20 percent, a correct prediction
was registered. If-one or more toxicity value and one
or more taxa values were over 20 percent, a correct
prediction was registered. This was done for all
stations and the correct prediction placed in the upper
left cell of the table. The same procedure was used for
each cell only changing the percentage to the
appropriate value for that cell. The 20 percent
incremental categories are arbitrarily selected.
The largest percentages of correct predictions were
obtained, in general, when comparable percentages
were compared, i.e., the highest values lie along a
diagonal from upper left to lower right. This pattern is
evidence that the degree of toxicity is related to the
degree of taxa reduction. To verify this trend quanti-
tatively, the degree of toxicity and reduction of taxa
was evaluated by a correlation analysis. The cor-
relation of the combined toxicity data (the greatest
toxicity of either the fathead minnows and the
Ceriodaphnia)and the reduction of the biological field
data (fish, zooplankton, and invertebrates) was signif-
icant (P <0.01). Figure 7-1 plots the greatest percent
toxicity at each station with the greatest reduction in
the field data that was subjected to the correlation
analysis.
One level of percent reduction or increase in toxicity
is not being proposed as the best percentage at this
time. Each study that has been done will compare
which reduction of the instream biological response
data best corresponds to a specified level of laboratory
toxicity. Comparisons for all sites studied need to be
completed before any decisions and recommendation
on the best percentage are made.
8.3 Summary
Ambient toxicity was measured at both stations
where effluent tests predicted toxicity. There was a
highly significant correlation between number of taxa
and degree of toxicity.
8-4
-------
CD
C
3
Percent Combined Toxicity
or
Combined Reduction in Taxa
N)
O
00
O
o
o
3
T3
a
•o
a
o
o>
3
3
0.
g.
1
(C
OC
lio
o •
? o
3 2
1 "
c <
3 D.
^ QJ
-------
References
Baumgardner, R K. 1966. Oxygen Balance in a
Stream Receiving Domestic and Oil Refinery Ef-
fluents. Ph D. Thesis, Oklahoma State University
70 pp
Hamilton, M. A. 1984. Statistical Analysis of the
Seven-Day Ceriodaphnia reticulata Reproductivity
Toxicity Test. EPA Contract J3905NASX-1. 16
January. 48 pp.
JRB Associates. 1983. Demonstration of the Site-
Specific Criteria Modification Process: Boggy and
Skeleton Creeks, Enid, Oklahoma. Draft Report
prepared for U.S EPA, Criteria of Standards
Division, Washington, D.C , EPA Contract 68-01-
6388.
Mount, D. l.andT. J. Norberg. 1984. A Seven-Day Life
Cycle Cladoceran Toxicity Test. Environ. Toxicol.
Chem. 3(3):425-434.
Mount, D. I. and T. J. Norberg. 1985. Validity of
Effluent and Ambient Toxicity for Predicting Bio-
logical Impact on Scippo Creek, Circleville, Ohio.
EPA Research Series, EPA/600/3-85/044.
Mount, D. I. and T. J. Norberg-Kmg. In press. Validity
of Effluent and Ambient Toxicity Testing for Predict-
ing Biological Impact on the Kanawha River,
Charleston, West Virginia EPA/600 Research
Series.
Mount, D. I., A. E. Steen, and T. J. Norberg-King, eds.
1985. Validity of Effluent and Ambient Toxicity for
Predicting Biological Impact on Five Mile Creek,
Birmingham, Alabama. EPA/600/8-85/01 5.
Mount, D. I., N. A. Thomas, T. J. Norberg, M T.
Barbour, T. H. Roush, and W. F. Brandes 1984
Effluent and Ambient Toxicity Testing and Instream
Community Response on the Ottawa River, Lima,
Ohio. EPA Research Series, EPA/600/3-84/084
Namminga, H. E. 1975. Heavy Metals in Water,
Sediments, and Chironomids in a Stream Receiving
Domestic and Oil Refinery Effluents. Ph.D. Thesis,
Oklahoma State University. 108 pp
Norberg, T. J. and D. I. Mount. 1 985. A New Fathead
Minnow (Pimephales promelas) Subchronic Tox-
icity Test. Environ. Toxicol. Chem. 4(5).
Robins, C. R., R. M. Bailey, C. E. Bond, J. R. Brooker, E.
A. Lachner, R. N. Lea, and W. B.Scott, eds. 1980. A
List of Common and Scientific Names of Fishes
from the United States and Canada. American
FisheriesSocietySpecial Publication No. 1 2, Fourth
Edition Committee on Names of Fishes, Bethesda,
Maryland. 174 pp.
Rogers, J 1 984. University of Wisconsin at Superior,
Wisconsin, and EPA Environmental Research Lab-
oratory at Duluth, Minnesota. July Personal com-
munication.
Sokal, R. R. and F. J. Rohlf 1981. Biometry W. H
Freeman and Company, New York.
Steele, G. R. and J H. Torrie. 1960. Principles and
Procedures of Statistics, a Bio-Metrical Approach
2nd Edition. McGraw-Hill, New York. 633 pp.
Wilhm, J. L. 1965 Species Diversity of Benthic
Macromvertebrates in a Stream Receiving Domes-
tic and Oil Refinery Effluents. Ph.D. Thesis, Okla-
homa State University 42 pp.
R-1
-------
Appendix A
Hydrological Sampling and Analytical Methods
A.1 Flow Measurements
Stream flows were measured from 9-11 August
using aTeledyne Gurley Pygmy flowmeter Measure-
ments were made once at Stations 1 through 9,
including an additional measurement downstream of
the Refinery At each station, measurements were
made at intervals of 0.3 to 0.6 m. depending on the
width of the transect such that a minimum of 10
velocity measurements were made
The water depth was recorded with each measure-
ment. Following standard hvdrological methods for
shallow streams (<0.75 m), velocity measurements
were made at depths of 60 percent of the water
column.
A.2 Flow Contribution Calculations
The mean contribution, m percent of the total flow,
was calculated using the measured stream velocities,
plant operating records, and USGS gauging station
data. The upstream flow values form Stations 1 and 2
for Boggy Creek and Skeleton Creek, respectively,
were used
A-1
-------
Appendix B
Toxicity Test and Analytical Methods
B.1 Sampling and Sample Preparation
A 24-hour composite sample of Refinery effluent
was collected 10-11 August 1983, as well as
composite samples of the stream stations. Automatic
ISCO samplers were set to collect an aliquot every 15
minutes and composite samples were collected in
5-gal polyethylene containers. The Fertilizer Plant
effluent was a partial composite and a partial grab
sample from the holding ponds on 10 August The
samples were cooled to approximately 10°C and
transported to ERL-Duluth where they were stored
until use at 8°C. Testing began 14 August 1 983. Test
solutions were renewed daily. Each day 2 L were
removed and warmed to 25°C. The effluent and the
dilution water were warmed separately, and dis-
solved oxygen levels checked for supersaturation
Ambient stations were also warmed to 25°C over a
propane heater and aerated until saturation was 100
percent.
The effluents were diluted with river water (Station
1A) that was collected upstream of the Refinery.
Dilutions were made using polypropylene or poly-
ethylene beakers and glass graduated cylinders Two
liters of each concentration were made and 0 200 L
were used for the Ceriodaphnia tests and the rest for
the fathead minnow tests. After the 2 L were
prepared, the dissovled oxygen (DO), pH, hardness,
and conductivity were measured. The DO and pH
meters were calibrated daily prior to readings At the
time of renewal, the DO was measured in one
compartment in each fathead minnow test chamber
(see Section B.3) and in at least one cup of the
Ceriodaphnia test in each exposure. DO was mea-
sured daily early in the morning after the lights were
on to evaluate any effects of diurnal DO cycles. DO
values m the 100 and 30 percent of the Fertilizer Plant
effluent were low, but otherwise no effects due to DO
levels were noticed. A series of effluent concentra-
tions of 100, 30,10, 3, and 1 percent were used m the
effluent dilution tests. For the ambient toxicity tests,
the samples were run without dilution.
B.2 Ceriodaphnia Test Method
Adult Ceriodaphnia from the ERL-Duluth culture
were used as brood stock, and the adults were not
acclimated m the dilution water prior to testing The
tests were started with less than 6-hour-old Cerio-
daphnia Glass beakers, 30-ml which contained 15
ml of test solution, were used Test solutions were
renewed daily and young, if present, were counted
and discarded. The animals were fed 0.05 ml of a
yeast food every day, for a concentration of 250 /JQ
yeast Temperatures were maintained at 25 r 1 °C by
means of a constant temperature cabinet The test
procedure was that of Mount & Norberg, 1 984.
B.3 Fathead Minnow Test Method
The methods used followed closely those described
by Norberg and Mount (1985) The test chambers
were 30.5 cm x 15.2 cm x 102 cm high, and are
divided into four compartments, this design allowed
four replicates for each concentration. The larval
fathead minnows were less than 24 hours old post
hatch and were from the ERL-Duluth culture The fish
were assigned to the test compartments by pipetting
1 or 2 fish at a time to each replicate test chamber
across all concentrations until all replicates had ten
fish in each or forty per concentration Newly hatched
brine shrimp were fed to the fish three times a day
The uneaten shrimp were removed daily by siphoning
the tanks during test solution renewal. At the same
time, the volume m the test chamber was drawn
down to 1 cm, after which 2 L of newtest solution was
added The laboratory temperature was 25 ± 1 °C A
16-hour light photopenod was used.
After seven days of exposure, the fish were preserved
m 4 percent formalin Prior to weighing, they were
rinsed in distilled water. Then each group was oven
dried for 18 hours m preweighed aluminum weigh
pans and weighed on a five-place analytical balance
B.4 Quantitative Analyses
B.4.1 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
B-1
-------
summed to derive the 7-day mean young value. By
this method, any young produced from females that
die during the test are included in the mean daily
estimate Using this procedure, mortalities of the
original females affect the estimate minimally, but
the mortality of the adult is used a long with the young
production to determine overall toxicity effects
Confidence intervals are calculated for the mean
reproductivity using a standard error estimate calc-
ulated by the bootstrap procedure The bootstrap
procedure subsamples the original dataset (1,000
times) by means of a computer to obtain a robust
estimate of standard error.
A Dunnett's two-tailed t-test is performed with the
effluent test data to compare each treatment to the
control for significant differences. For the ambient
station data, Tukey's Honestly Significant Difference
Test is used to compare stations.
8.4.2 Fathead Minnows
The four groups' mean weights are statistically
analyzed with the assumption that the four test
chamber compartments behave as replicates. The
method of analysis assumes the variability in the
mean treatment response is proportional to the
number of fish per treatment. MINITAB (copyright
Pennsylvania State University 1982) was used to
estimate a t-statistic for comparing the mean treat-
ment 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 1 960) The survival
data are arcsine-transformed prior to the regression
analyses to stabilize variances for percent data to
show significant differences; however, actual survival
values of the replicates are given in Tables 4-3 and
4-6.
B-2
-------
Appendix C
Biological Sampling and Analytical Methods
Estimated pool and riffle proportions and percent
cover information are provided in Table C-1. Table 2-1
provides information on which stations were sampled
for each survey.
C.1 Plankton Survey
Plankton were collected from ten stations on Skeleton
Creek and Boggy Creek near Enid, Oklahoma, on 8-11
August 1983. Duplicate samples were collected at
each station using a Wisconsin-type plankton net
with a 80-/um mesh. The net was held stationary in
the water for two minutes (only one minute at Station
9). The samples were transferred to bottles pre-
charged with formalin. The volume filtered was
calculated the time required for afloat to travel a 3-m
distance and the net diameter, assuming 100 percent
filtering efficiency.
The samples were thoroughly mixed and an aliquot
removed. Two subsamples from each replicate sam-
ple were analyzed using a Sedgewick-Rafter counting
chamber. Identifications were made using a com-
pound microscope at 100X magnification. All organ-
isms in the chamber were enumerated and identified
to a convenient taxon, except the solitary diatoms. For
diatoms, one short-dimension optical strip was
enumerated. Abundance was standardizedto number
per liter for density comparisons.
The crustacean and rotifer densities were analyzed by
Analysis of Variance (ANOVA) One-way ANOVAs
were performed to determine differences between
stations. Tukey's Honestly Significant Difference
(HSD) tests were conducted to determine which
stations were different when a significant difference
was detected using the ANOVAs
C.2 Macroinvertebrate Survey
C.2.1 Sample Collection
C.2.1.1 Natural Substrates
Natural substrates at ten stations were sampled on
Skeleton Creek and Boggy Creek from 8-11 August
1 983. A 1 -ft2 Hess-style sampler was used with a 800
x900-um mesh net. Triplicate samples were collected
in riffle areas or similar areas and then preserved in
10 percent formalin.
Table C-1. Station Description Information and Estimated Proportions of Riffle and Pool for Skeleton Creek and Boggy Creek,
Enid, Oklahoma
Station
1A
1
2
3
4
5
5A
6
7
8
9
10
Estimated
Percent
Cover
100
100
90
80
0
10
0
0
0
0
20
0
Percent
Riffle
16
0
40b
50b
100b
40
23
100"
27
16
27
0
Riffle
Width
(m)
45
C
09
6 1
91
46
30
122
6 1
6 1
122
C
Riffle
Length
(m)
6 1
C
6 1
9 1
305
6 1
9 1
305
9 1
6 1
9 1
C
Percent
Pool
84
100
60
50
0
60
77
0
73
84
73
100
Pool
Width
(m)
6 1
C
36
9 1
91
91
122
137
122
107
Pool
Length8
(m)
30
C
9 1
18 2
152
305
--
244
305
244
C
Estimated
area (m2) for
Fish Seme
210
C
106
191
214
106
214
210
171
214
171
C
"Estimated sampled length for the fish survey, actual length may be longer
"Run habitat only.
°These stations sampled with artificial substrates only
C-1
-------
C.2.1.2 Artificial Substrates
Quadruplicate Hester-Dendy muItiplate artificial sub-
strates were suspended m the water column at each
of the ten stream stations on 20 July 1983 and
removed on 9 August, resulting in a 20-day colon-
ization period Substrates were collected using a
small-mesh net Each substrate was disassembled
and scraped and the col lections were preserved in 1 0
percent formalin The Hester-Dendy substrates have
an effective surface area of 0 093 m2
C.2.2 Sample Analysis
The samples were washed m tap water and flooded
with a sugar solution to separate debris and organ-
isms. The floating organisms were removed and
placed m 70 percent ethanol The debris was exam-
ined to detect non-floating or entangled organisms
using a dissecting microscope at 8X magnification
Organisms were enumerated and identified to genus
or lowest reasonable taxa Abundance was stand-
ardized to number per square meter for density
comparisons
ANOVAs were conducted on the counts of major taxa
to determine differences between stations. The two
major taxa were Dicrotendipes sp. and Berosus sp.
Tukey's HSD tests were conducted when significance
was detected using ANOVA, to determine which
stations were different. In addition, ANOVAs were
conducted on the number of Chironomidae taxa and
the total number of macroinvertebrate taxa to discern
differences between stations. Tukey's HSD was used
when the ANOVAs showed significant differences to
identify which of the stations were different.
C.3 Fish Survey
Fish seining was done at ten stations on Skeleton
Creek and Boggy Creek on 8-11 August 1983. At
most, 30.5 m of the stream was seined at each station
using a woven net, 1.2 m x 9.1 m, with a 0 5-cm
mesh. Collections were preserved in 10 percent
formalin. Fish were enumerated and identified to
species or lowest practicable taxon
The number of fish taxa per station were examined
using a X2 test This test was performed with Station
1A as the expected value, and again with Station 2 as
the expected value.
C-2
-------
Appendix D
Biological Data
Table D-1.
Mean Density (No
Boggy Creek, Enid
,/m2) of Benthic Macroinvertebrates Collected from Natural Substrates in Skeleton Creek and
, Oklahoma, August 1983
Sampling Station
Taxa
Ephemeroptera
Caenis sp
Tricorythodes sp
Baetis sp
Stenonema sp
Choroterpes sp
Total Ephemeroptera
Trichoptera
Hydroptihdae
Cheumatopsyche sp
Hydropsyche sp
Hydropsychidae pupae
Total Trichoptera
Coleoptera
Laccophilus sp
Pe/todytes sp
Berosus sp
Stenelmis sp
Dubiraphia sp
Total Coleoptera
Odonata
Dromogomphus sp
Gomphus sp
Plathemis sp
Libellula sp
Argia sp
Total Odonata
Megaloptera
Corydalis sp
Neohermes sp
Total Megaloptera
Diptera
Chironomus sp
Dicrotendipes sp
Polypedilum sp
Cryptochironomus sp
Pseudochironomus sp
K/efferulus sp
Tanytarsus sp
Micropsectra sp
Cricotopus sp
Psectrocladius sp
Ablabesmyia sp
Pentaneura sp
T any pus sp.
Procladius sp.
Chironomidae pupae
1A
251
7
11
--
269
29
14
--
43
90
14
--
104
--
--
--
90
90
--
--
0
14
352
154
--
7
--
90
25
14
--
65
11
4
--
32
2
7
4
--
--
--
11
--
--
--
0
--
--
29
--
--
29
--
0
--
0
--
298
43
22
102
22
--
43
4
--
7
--
--
3
14
--
14
--
--
0
11
--
--
11
--
0
--
0
488
466
18
7
--
--
104
--
7
--
11
--
32
--
61
4
--
--
--
0
--
--
0
4
459
--
--
463
--
22
4
26
0
513
553
14
4
--
--
387
--
--
79
606
29
25
5
29
11
--
--
--
40
--
4
4
4
--
126
7
4
141
4
--
4
0
703
1,568
39
4
--
25
50
--
434
4
248
22
811
5A
39
79
--
--
--
118
--
--
--
--
0
--
--
1,159
--
--
1,159
--
--
--
4
4
0
355
621
14
--
--
29
47
--
176
--
111
50
305
6
11
86
--
97
4
4
--
8
--
--
861
4
865
--
--
--
29
29
--
0
4
312
14
--
4
14
11
--
240
7
79
--
--
147
7
39
--
36
75
29
--
--
29
--
--
947
947
--
--
0
--
4
4
11
240
4
4
--
43
352
258
4
--
90
8
--
--
1 1
11
104
7
--
111
--
--
151
151
--
--
0
--
0
122
746
11
11
--
402
197
--
196
--
194
9
--
86
197
7
29
319
1,137
273
22
1,432
--
--
32
18
50
--
1 1
--
1 1
22
22
7
481
4
--
36
4
--
25
--
47
D-1
-------
Table D-1 . (Continued)
Taxa 1A 2 3
Palpomia sp
Probezzia sp 11147
Simuhdae 4
Diptera pupae3 -- 7
Total Diptera 783 562 1,201
Hemiptera
Belastoma sp
Conxidae
Total Hemiptera 000
Others
Gastropoda
Physidae 29 50 4
Ancyhdae 32
Pelecypoda
Sphaerndae 4
Amphipoda
Tahtridae 7
Oligochaeta (unidentified) 47 93 104
Annelida
Hirudinea
Total number of taxa" 25 14 13
Total number of mdividuals/m2 1,408 745 1,334
"Unidentified, non-Chironomidae pupae
"Does not include pupae
Note Values are rounded to nearest integer
Table D-2. Mean Density (No./m2) of Macroinvertebrates
Creek, Enid, Oklahoma, August 1983
Taxa 1 2 3
Ephemeroptera
Caenis sp 1 26 70 1 1
Tncorythodes sp
Baetis sp 8 54 1 6
Stenonema sp --83
Choroterpes sp
Total Ephemeroptera 134 132 30
Trichoptera
Chimarra sp
Hydrotilidae
Cheumatopsyche sp
Hydropsyche sp
Hydropsychidae pupae
Total Trichoptera 000
Coleoptera
Tropisternus sp
Laccophilus sp
Berosus sp. -- 73 13
Stenelmis sp --58
Total Coleoptera 0 78 21
Odonata
Libellula sp -- 3
Argiasp 38 234 177
Hetaenna sp
Ischnura sp. 40
Total Odonata 38 277 177
Sampling Station
4 5 5A 6
__
65 4 4 -
2,275 3,912 1,712 832
4
61 32 68
0 61 32 72
208 97 11 47
4
18
39 4 4
14 36 814 169
7
16 23 16 24
3,025 4,299 3,850 2,152
Collected from Artificial Substrates
Sampling Stations
4 5 6a 7
27 11 11 65
65 65 226 1,043
92 76 237 1,108
4
0040
-- 78
7
207 135 1,387 1,755
11 -- -- 5
218 135 1,401 1,768
3
183 22 143 290
11
43 3 22 5
227 25 165 306
7 8
4
4
1,014 1,879
..
4
4 0
129 22
__
11 215
18 13
2,213 2,389
in Skeleton Creek
8 9
..
19 75
188 126
5 16
8
220 217
13
3
2,027 145
554 1,473
70 86
2,667 1,704
--
654
108 46
762 46
118 132
118 132
9
__
--
--
604
--
--
0
7
--
-.
158
--
19
2,603
and Boggy
10
3
3
3
9
--
--
75
3
78
--
--
0
140
140
D-2
-------
Table D-2. (Continued)
Sampling Stations
Taxa
Megaloptera
Chauliodes sp
Neohermes sp
Total Megaloptera
Diptera
Chironomis sp
Dicrotendipes sp
Polypedilum sp.
Cryptochironomus sp
Pseudochironomus sp
K/efferu/us sp
Tanytarsus sp.
Tnbelus sp
Cricotopus sp.
Psectrocladius sp.
Ablabesmyia sp
Pentaneura sp.
Corynoneura sp
Tanypus sp.
Procladius sp.
Chironomidae pupae
Palpomia sp
Probezzia sp.
Tabannus sp
Athenx sp
Hemerodromia sp.
Total Diptera
Hemiptera
Belastoma sp.
Conxidae
Total Hemiptera
Others
Gastropoda
Physidae
Amphipoda
Talitridae
Oligochaeta
Annelidae
Hirudinea
Total Number of Taxa"
Total Number of Individuals/m2
1
--
--
0
99
1,038
11
--
13
--
83
--
--
--
253
8
13
--
78
--
5
--
--
--
1,601
--
--
0
--
16
--
--
13
1,789
2
--
--
0
13
2,368
27
--
8
3
89
--
--
--
250
16
--
54
8
353
27
8
--
--
--
3,224
--
--
0
226
--
--
--
21
3,717
3
--
59
59
212
2,640
43
3
--
5
3
--
99
314
1,938
11
--
5
--
260
--
--
--
--
5,533
--
--
0
11
40
--
20
5,871
4
24
24
35
1,790
5
--
--
--
3
--
11
81
441
22
--
75
19
75
--
--
--
--
2,557
--
--
0
427
223
8
--
21
3,356
5
3
3
631
1,352
5
--
--
357
5
--
56
22
653
3
--
454
--
505
--
3
--
--
4,046
--
--
0
13
32
--
5
20
4,325
6a
--
7
7
143
190
4
7
--
--
14
--
36
36
229
--
--
36
65
--
--
--
--
--
764
4
7
11
25
32
--
23
2,650
7
--
8
8
667
1,691
8
11
38
--
27
--
19
186
267
--
--
164
--
56
3
--
3
--
--
3,140
--
3
3
13
161
--
8
25
6,505
8
13
32
45
5
83
6,024
5
--
--
3,255
--
5
124
728
30
--
--
--
817
3
3
--
3
3
1 1 ,088
--
0
32
19
--
--
28
14,951
9
--
--
0
3
126
1,769
--
48
226
--
48
40
204
--
13
3
--
148
--
--
--
--
--
2,628
--
3
3
--
--
18
4,730
10
--
0
--
30
202
--
30
8
24
3
3
153
--
--
--
--
118
--
--
--
--
--
571
--
--
0
--
--
13
798
"Station 6 had only three replicates.
NOTE: Values are rounded to nearest integer
Table D-3. Analysis of Variance and Tukey's Studentized Range Test Results for Zooplankton, Skeleton Creek, August 1983
Crustaceans
Dependent Variable: In Count
Source
Station
Error
Corrected total
dF
9
30
39
Sum of
Squares
2767
5.44
33 12
Mean
Square
3.07
0 18
F Value PR > F
1693 0.0001
Tukey's Studentized Range Test
Station
Mean
7 3
2 44 2.06
8 4
055 0.44
6 2
0.34 0.24
1A 9 5A 5
015 0 05 0.05 0 0
D-3
-------
Table D-3. (Continued)
Dependent Variable In Count
Rotifers
Source
Station
Error
Corrected total
dF
9
30
30
Sum of
Squares
2985 23
5030
3035.53
Mean
Square
331 69
1 68
F Value
19782
PR >F
00001
Station
Mean
7
3064
6
1459
Tukey's Studentized Range Test
5A
1057
5
780
4
5 18
3
5 13
374
9
239
2
047
1A
0 18
Table D-4. Analysis of Variance and Tukey's Studentized Range Test Results for the Two Most Abundant Macroinvertebrate
Taxa from the Natural Substrates, Skeleton Creek, August 1983
Dependent Variable Count
Dicrotendipes sp
Source
dF
Sum of
Squares
Mean
Square
F Value
PR >F
Station
Error
Corrected total
9
20
29
3396
1473
48.69
377
074
5 12
00001
Station
Mean
5
478
5A
400
Tukey's Studentized Range Test
3
361
6
3 39
4
3 15
2
304
1A
303
7
303
8
2 50
9
046
Dependent Variable Count
Berosus sp
Source
Station
Error
Corrected total
dF
9
20
29
Sum of
Squares
51 30
14 19
65.49
Mean
Square
570
071
F Value
803
PR > F
00001
Tukey's Studentized Range Test
Station
Mean
7
440
6
438
5A
4 12
4
331
5
2 52
8
249
1A
1 94
9
1 36
2
1 23
3
060
D-4
-------
Table D 5. Analysis of Variance and Tukey's Studentized Range Test Results for the Two Most Abundant Macroinvertebrate
Taxa from the Artificial Substrates, Skeleton Creek, August 1983
Dicrotendipes sp
Dependent Variable Count
Source dF
Station 9
Error 29
Corrected total 38
Station 3 2
Mean 24550 22025
Dependent Variable Count
Source dF
Station 9
Error 29
Corrected total 38
Station 7 6
Mean 16325 12933
Table D-6. Analysis of Variance and Tukey
Creek, August 1983
Natural Substrate Data
Dependent Variable Count
Source dF
Station 9
Error 20
Corrected total 29
Station 1A 5
Mean 1700 1567
Dependent Variable' Count
Source dF
Station 20
Error 9
Corrected total 29
Sum of Mean
Squares Square F Value
297,23927 33,02658 477
200,67017 6,91667
497,90944
Tukey's Studentized Range Test
4751 69
16650 15725 12575 9650 1767 1175
Berosus sp
Sum of Mean
Squares Square F Value
120,450.58 13,38340 6426
6,03942 20825
126,49000
Tukey's Studentized Range Test
845231
6075 1925 12 50 675 1 25 00
PR >F
00006
8 10
775 275
PR >F
00001
9 10
00 00
's Studentized Range Test Results for Numbers of Macroinvertebrate Taxa, Skeleton
Total Number of Taxa
Sum of Mean
Squares Square F Value
17487 1944 374
104.00 520
278.97
Tukey's Studentized Range Test
6 9 5A 7 4 8
15.00 1433 1333 1267 1200 1100
Total Number of Chironomidae Taxa
Sum of Mean
Squares Square F Value
2883 320 1 48
43 43 217
72 17
PR >F
00067
2 3
1067 867
PR >F
02224
Tukey's Studentized Range Test was not performed since the ANOVA results were nonsignificant
D-5
-------
Table D-6. (Continued)
Artificial Substrates
Dependent Variable Count
Total Number of Taxa
Source
Station
Error
Corrected total
dF
9
29
38
Sum of
Squares
38033
11742
49775
Mean
Square F Value
4226 1044
404
PR >F
00001
Tukey's Student/zed Range Test
Station
Mean
8 7
1875 1725
4 6
1675 1667
2935
1550 1475 1400 1350
1 10
975 850
Total Number olChironomidae Taxa
Dependent Variable Count
Source
Station
Error
Corrected total
dF
9
29
38
Sum of
Squares
31 06
4392
7492
Mean
Square
344
1.51
F Value PR > F
227 00453
Tukey's Student/zed Range Test
Station
Mean
7 4
875 825
5 6
775 767
9 3
750 725
2 8 1 10
675 650 625 575
Table D-7. List of Fish Species and Families Collected from Skeleton Creek and Boggy Creek Near Enid, Oklahoma9
Family
Scientific Name
Common Name
Cypnmdae
(minnows)
Centrarchidae
(sunfish)
Notropis lutrensis
Notropis stramineus
Notropis umbratilis
Pimephales promelas
Phenacobis mirabilis
Notemigonus crysoleucas
Notropis spp
Lepomis megalotis
Red shiner
Sand shiner
Redfm shiner
Fathead minnow
Suckermouth minnow
Golden shiner
Early juvenile cyprmids
Longear sunfish
Ictalundae
(catfish)
Poecilndae
(livebearers)
Catastomidae
(suckers)
Lepomis cyanellus
Lepomis humi/us
Ictalurus me/as
Gambusia aff/nis
Carpoides spp
Green sunfish
Orangespotted sunfish
Black bullhead
Mosquitofish
Early juvenile catastomids
"Names follow Robins et al (1980)
D-6
------- |