EPA/600/8-86/001
July 1986
The Validity of Effluent and
Ambient Toxicity Tests for
Predicting Biological Impact,
Back River, Baltimore Harbor, Maryland
Edited by
Donald I. Mount, Ph.D.1
Alexis E. Steen2
Teresa Norberg-King1
'Environmental Research Laboratory
U.S. Environmental Protection Agency
6201 Congdon Blvd.
Duluth, Minnesota 55804
2EA Engineering Science, and Technology, Inc.
Hunt Valley/Loveton Center
15 Loveton Circle
Sparks, Maryland 21152
Environmental Research Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Duluth, MISI 55804
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Disclaimer
This document has been reviewed in accordance with U.S. Environmental
Protection Agency policy and approved for publication. Mention of trade names
or commercial products does not constitute endorsement or recommendation
for use.
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Foreword
The Complex Effluent Toxicity Testing Program was initiated to support the
developing trend toward water quality-based toxicity control in the National
Pollutant Discharge Elimination System (NPDES) permit program. It is designed
to investigate, under actual discharge situations, the appropriateness and utility
of "whole effluent toxicity" testing in the identification, analysis, and control of
adverse water quality impact caused by the discharge of toxic effluents.
The four objectives of the Complex Effluent Testing Program are:
1. To investigate the validity of effluent toxicity tests in predicting adverse
impact on receiving waters caused by the discharge of toxic effluents.
2. To determine appropriate testing procedures which will support regulatory
agencies as they begin to establish water quality-based toxicity control
programs.
3. To provide practical case examples of how such testing procedures can be
applied in different toxic effluent discharge situations involving discharges
to a variety of discharge situations.
4. To field test short-term chronic toxicity tests including the test organisms,
Ceriodaphnia dubia and Pimphales promelas.
Until recently, NPDES permitting has focused on achieving technology-based
control levels for toxic and conventional pollutants in which regulatory
authorities set permit limits on the basis of national guidelines. Control levels
reflected the best treatment technology available, considering technical and
economic achievability. Such limits did not, nor were they designed to, protect
water quality on a site-specific basis.
The NPDES permits program, in existence for over 10 years, has achieved the
goal of implementing technology-based controls. With these controls largely in
place, future controls for toxic pollutants will, of necessity, be based on site-
specific water quality considerations.
Setting water quality-based controls for toxicity can be accomplished in two
ways. The first is the pollutant-specific approach which involves setting limits for
single chemicals, based on laboratory-derived no-effect levels. The second is the
"whole effluent" approach which involves setting limits using effluent toxicity
as a control parameter. There are advantages and disadvantages to both
approaches.
The "whole effluent" approach eliminates the need to specify a limit for each of
thousands of substances that may be found in an effluent. It also includes all
interactions between constituents as well as biological availability.
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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 Virignia
8. Kanawha River, Charleston, West Virginia
This report presents the site study on Back River, Baltimore Harbor, Maryland,
which was conducted in March 1984. The study site was an estuary of the
Chesapeake Bay and receives discharges including a large POTW discharge.
This report presents the site study on Back River, Baltimore Harbor, Maryland,
issuance or enforcement activities.
Rick Brandes
Permits Division
Nelson Thomas
ERL/Duluth
Project Officers
Complex Effluent Toxicity
Testing Program
IV
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Contents
Page
Foreword iii
List of Figures vii
List of Tables ix
Acknowledgments xii
List of Contributors xiii
Executive Summary xiv
Quality Assurance xv
1. Introduction 1-1
2. Study Design 2-1
2.1 Toxicity Testing Study Design 2-1
2.2 Hydrological Survey Study Design 2-1
2.3 Biological Survey Study Design 2-1
2.4 Integration of Laboratory and Field Efforts 2-1
2.5 Research on Effluent Fractionation 2-1
3. Site Description 3-1
4. Toxicity of Effluents and Receiving Water 4-1
4.1 Chemical and Physical Test Conditions 4-1
4.2 Results of Fathead Minnow Growth Tests 4-1
4.3 Results of Ceriodaphnia Reproduction Potential Tests 4-4
4.4 Results of the Microtox® Tests 4-6
4.5 Summary of Toxicity Data 4-7
5. Hydrological Studies of Patapsco River 5-1
5.1 Dilution Analysis of the Patapsco POTW 5-1
5.2 Evaluation of Hydrological Conditions of the Patapsco River ... 5-1
6. Hydrological Studies of Back River 6-1
6.1 Dilution Analysis of the Back River POTW 6-1
6.2 Hydrological Modeling of Back River 6-1
6.3 Evaluation of Hydrological Conditions of the Back River
and Middle River 6-1
7. Macrozooplankton/lchthyoplankton of Back River and
Middle River 7-1
7.1 Community Structure 7-1
7.1.1 Macrozooplankton 7-1
7.1.2 Ichthyoplankton 7.1
7.2 Differences Between Stations in Key Macrozooplankton
Taxa - 7-1
7.3 Evaluation of the Macrozooplankton Community 7-3
8. Benthic Macroinvertebrates of Back River and Middle River 8-1
8.1 Community Structure 8-1
8.2 Spatial Trends in Key Taxa 8-1
8.3 Evaluation of the Benthos Community 8-3
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Contents (cont'd)
Page
9 Fish Community Survey 9-1
9.1 Community Structure 9-1
9.2 Fish Condition 9-1
9.3 Evaluation of the Fish Community 9-3
10. Comparison of Laboratory Toxicity Data and Receiving Water
Biological Impact 10-1
11. Effluent Fractionation Testing 11-1
11.1 Fractionation Test Results 11-1
11.1.1 Ceriodaphnia 48-Hour Acute Tests 11-1
11.1.2 Microtox® Tests 11-2
11.1.3 Chemical Analyses of Toxic Fractions 11-3
11.2 Summary 11-3
References R-1
Appendix A: Toxicity Tests and Analytical Methods A-1
A. 1 Sampling and Sample Preparation A-1
A.2 Ceriodaphnia Test Methods A-1
A.3 Fathead Minnow Test Method A-2
A.4 Ceriodaphnia Statistical Analysis A-2
A.5 Fathead Minnow Statistical Analysis A-2
A.6 Microtox® Testing Methods A-2
Appendix B: Hydrological Sampling and Analytical Methods B-1
B.1 Patapsco River Survey B-1
B.2 Back River and Middle River B-1
Appendix C: Biological Survey Sampling and Analytical Methods C-1
C.1 Plankton Survey C-1
C.2 Benthic Macroinvertebrate Survey C-1
C.3 Fish Survey C-1
Appendix D: Effluent Fractionation and Toxicity Testing Methods D-1
D.I Sampling D-1
D.2 Ceriodaphnia Culture, Maintenance and Testing D-1
D.3 Microtox® D-1
D.4 Chemical Fractionation D-1
Appendix E: Toxicity Test Data E-1
Appendix F: Biological Data F-1
Appendix G: Support Chemical Fractionation Data G-1
VI
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List of Figures
Number Title Page
3-1 Study area showing the two wastewater treatment plants and
the biological sampling stations in Back River, Middle
River, and Patapsco River 3-1
5-1 Surface dilution contours at the Patapsco POTW, 1103 through
1217 hours, 22 March 1984 5-1
5-2 Surface dilution contours of the Patpasco POTW, 1 238 through
1417 hours, 22 March 1 984. Also shown are locations of
vertical sampling stations 5-1
6-1 Dye concentrations in the Back River as observed and predicted
by the numerical model. Dye injection started at hour 62 6-2
6-2 Dye concentration in the Back River as predicted by the
numerical model for simulated dye release beginning
1 March 6-2
7-1 Spatial trends of macrozooplankton community parameters,
March 1 984 7-2
8-1 Spatial trends of benthic community parameters 8-3
9-1 Spatial comparison of fish catches in Back River and Middle
River on two days in March 1 984 9-3
11-1 Schematic results of Ceriodaphnia acute tests on effluent
fractions 11-2
11 -2 Schematic results of Microtox® tests on effluent fractions 11-3
B-1 Map showing the Back River segmentation scheme and water
sampling locations B-2
D-1 Fractionation and testing procedure D-2
G-1 Base/neutrals standard reconstructed ion chromatogram for
3-day and 7-day Patapsco POTW base/neutral fraction
effluent analysis G-8
G-2 Surrogate spike standard reconstructed ion chromatogram
for 3-day and 7-day Patapsco POTW base/neutral fraction
effluent analysis G-9
G-3 Blank reconstructed ion chromatogram for 3-day and 7-day
Patapsco POTW base/neutral fraction effluent analysis G-10
G-4 Reconstructed ion chromatogram for 3-day Patapsco
POTW base/neutral fraction effluent analysis G-11
G-5 Library search for possible compound from 3-day Patapsco
POTW base/neutral fraction effluent analysis G-1 2
G-6 Reconstruction ion chromatogram for 7-day Patapsco
base/neutral fraction effluent analysis G-13
vii
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List of Figures (cont'd)
Number Title Page
G-7 DFTPP reconstructed ion chromatogram for 3-day and 7-day
Patapsco POTW base/neutral fraction effluent
analysis G-14
G-8 DFTPP mass spectrum for 3-day and 7-day Patapsco
base/neutral POTW fraction effluent analysis G-1 5
VIII
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List of Tables
Number Title Page
4-1 Seven-Day Percent Survival of Larval Fathead Minnows
Exposed to Various Concentrations of Effluents in
Reconstituted Water, Baltimore Harbor, Maryland 4-2
4-2 Mean Weight of Larval Fathead Minnows Exposed to Various
Concentrations of Effluents in Reconstituted Water,
Baltimore Harbor, Maryland 4-2
4-3 Seven-Day Percent Survival of Larval Fathead Minnows
Exposed to Waters from Various Stream Stations for
Ambient Toxicity, Baltimore Harbor, Maryland 4-3
4-4 Mean Weight of Larval Fathead Minnows Exposed to Waters
from Various Stations for Ambient Toxicity, Baltimore Harbor,
Maryland 4-3
4-5 Seven-Day Mean Percent Survival and Weight of Larval
Fathead Minnows for Salinity Test at Baltimore Harbor,
Maryland 4-4
4-6 Daily and Mean Salinity at Back River Stations, Baltimore
Harbor, Maryland 4-4
4-7 Daily and Seven-Day Effluent Concentrations at Back River
Stations, Baltimore Harbor, Maryland 4-5
4-8 Reproduction and Survival of Ceriodaphnia dubia for the
Patapsco POTW Effluent and the Patapsco and Middle Rivers
Ambient Stations, Baltimore Harbor, Maryland 4-5
4-9 Daily Survival and Mean Young Production of Ceriodaphnia
dubia in Various Dilutions of Back River POTW Effluent,
Baltimore Harbor, Maryland 4-5
4-10 Daily Survival and Mean Young Production of Ceriodaphnia
dubia in Back River Ambient Station Water, Baltimore
Harbor, Maryland 4-6
4-11 Daily Survival and Mean Number of Young Per Female in the
Salinity Test, Baltimore Harbor, Maryland 4-6
4-12 EC50 Values for Microtox® Test for the Two POTW
Effluents, Baltimore Harbor, Maryland 4-6
4-13 1 5-Minute Percent Light Reduction for 91 Percent Back River
Ambient Samples, Baltimore Harbor, Maryland 4-7
5-1 Vertical Measurements of Dilution at Stations Surrounding
Patapsco POTW Discharge, Baltimore Harbor, March 1984 5-2
6-1 Surface Water Quality Data for Back River and Middle River
Stations from 9 March 1984 Through 16 March 1984 6-3
7-1 Abundance and Percent Composition of the Macrozooplankton
Community of Back River and Middle River, 12 March 1984 7-1
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List of Tables fcont'd)
Number Title Page
7-2 Abundance and Percent Composition of the Macrozooplankton
Community of Back River and Middle River, 1 6 March 1 984 .... 7-1
7-3 Composition of the Macrozooplankton Community of Back
River and Middle River, 12 and 1 6 March 1 984 7-2
8-1 Abundance of Benthic Macroinvertebrates Collected from
Back River and Middle River, 19 March 1984 8-2
8-2 Composition of Benthic Community of Back River and
Middle River, 1 9 March 1 984 8-3
8-3 Shannon-Wiener Diversity Indices Associated Evenness and
Redundancy Values, and Community Loss Index Calculated
'on Benthic Data from Back River and Middle River,
19 March 1 984 8-4
9-1 Fish Catch and Water Quality Parameters, in Back River
and Middle River, 7 March 1 984 9-2
9-2 Fish Catch and Water Quality Parameters, in Back River
and Middle River, 14 March 1984 9-2
9-3 Observations of Abnormalities by Species in Back River
and Middle River, 7 March 1 984 9-4
9-4 Observations of Abnormalities by Species in Back River
and Middle River, 14 March 1984 9-4
11-1 LC50 Values Calculated by Moving Average Method, Based
on Cenodaphnia dub/a 48-Hour Acute Tests 11-1
11 -2 EC50 Values Based on Beckman Microtox® Acute Tests 11-3
11 -3 Levels of Pesticides, Herbicides, and PCBs m 3-Day and
7-Day Composite Patapsco POTW Effluents 11-4
1 1-4 Levels of Base/Neutral Compounds, Determined by GC/MS
Analysis (EPA Method 625), for 3-Day and 7-Day Patapsco
POTW Effluents 11-4
B-1 Dye Plume Mappings B-3
D-1 Formulation for Moderately Hard Reconstituted Water and
Final Water Quality Ranges D-1
E-1 Routine Chemistry Data for the Ambient Tests, Baltimore
Harbor, Maryland E-1
E-2 Routine Chemistry Data for the Effluent Dilution and Salinity
Test E-1
E-3 Final Dissolved Oxygen Levels for Ceriodaphnia dubia Effluent,
Ambient, and Salinity Tests, Baltimore Harbor, Maryland E-2
F-1 Results of a X2 Test Performed on the Number of
Macrozooplankton Taxa, Back River, March 1984 F-1
F-2 Abundance of Macrozooplankton Collected from Back River
and Middle River, 12 March 1984 F-1
F-3 Abundance of Macrozooplankton Collected from Back River
and Middle River, 1 6 March 1 984 F-2
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List of Tables (cont'd)
Number Title Page
F-4 Analysis of Variance and Tukey's Studentized Range Test
Results for Eurytemora affinis, Back River, March 1984 F-2
F-5 Water Quality Data from Back River and Middle River, 12 and
16 March 1984 F-3
F-6 Results of a X2 Test Performed on the Number of Benthic
Macroinvertebrate Taxa, Back River, March 1984 F-4
F-7 Water Quality Data from Back River and Middle River, 16 March
1984 F-4
F-8 Results of a X2 Test Performed on the Number of Fish
Taxa, Back River, March 1984 F-4
F-9 Trends in Abnormalities Observed Among Brown Bullheads
Collected in Back River and Middle River, 7 March 1984 F-5
F-10 Trends in Abnormalities Observed Among Brown Bullheads
Collected in Back River and Middle River, 14 March 1984 F-5
F-11 Trends in Abnormalities Observed Among White Perch
Collected in Back River and Middle River, 7 March 1984 F-6
F-12 Trends in Abnormalities Observed Among Pumpkinseed and
White Perch Collected in Back River and Middle River,
14 March 1 984 F-5
F-13 List of Fish Species and Families Collected, Back River
and Middle River, March 1 984 F-7
G-1 Ceriodaphnia dubia Mortality in 48-Hour LC50 Test^ on Back
River and Patapsco POTW G-2
G-2 Base/Neutral Standard Quantitation Report for 3-Day and
7-Day Patapsco POTW Base/Neutral Fraction Effluent
Analysis G-3
G-3 Surrogate Spike Standard Quantitation Report for 3-Day and
7-Day Patapsco POTW Base/Neutral Fraction Effluent
Analysis G-3
G-4 Blank Quantitation Report for 3-Day and 7-Day Patapsco
POTW Base/Neutral Fraction Effluent Analysis G-4
G-5 Spike Blank Quantitation Report for 3-Day and 7-Day Patapsco
POTW Base/Neutral Fraction Effluent Analysis G-4
G-6 Quantitation Report for 3-Day Patapsco POTW Base/Neutral
Fraction Effluent Analysis G-5
G-7 Quantitation Report for 7-Day Patapsco POTW Base/Neutral
Fraction Effluent Analysis G-6
G-8 Mass List for DFTPP Analysis on 3-Day and 7-Day Patapsco
POTW Base/Neutral Fraction Effluent G-7
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A ckno wledgments
The assistance of Robert Donaghy and Robert Green from EPA Region III in
conducting the onsite Microtox® tests was appreciated. The aid of Floyd
Boettcher, Environmental Research Laboratory—Duluth, as field engineer is
gratefully acknowledged.
XII
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List of Contributors
LABORATORY TOXICITY TESTS
Donald I. Mount1, Scott E. Heinritz1, and Jeffrey S. Denny'
HYDROLOGICAL SURVEY
Stephen R. Rives2
PLANKTON SURVEYS
Richard A. Connelly2 and Michael T. Barbour2
BENTHIC MACROINVERTEBRATE SURVEY
Richard A. Connelly2 and Michael T. Barbour2
FISH SURVEYS
Michael D. Schmitt2
COMPARISION OF LABORATORY TOXICITY DATA AND
RECEIVING WATER BIOLOGICAL IMPACT
Donald A. Mount1
EFFLUENT FRACTIONATION TESTING
Wayne C. McCulloch2 and Stephen E. Storms2
PRINCIPAL INVESTIGATOR
Donald I. Mount, Ph.D.'
'U S. Environmental Protection Agency, Environmental Research Laboratory—Duluth, 6201 Congdon Blvd.,
Duluth, MN 55804
2EA Engineering, Science, and Technology, Inc., Hunt Valley/Loveton Center, 15 Loveton Circle, Sparks, MD
21152
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Executive Summary
The toxicity of freshwater effluents discharged to brackish waters are difficult to
assess because of the role of salinity. If high concentrations of effluents are of
concern, then freshwater organisms are better since salinity will be low. If low
concentrations are of concern then brackish water species are better for testing.
The purpose of this study was to measure the toxicity of effluents discharged to
an estuary using freshwater test species and compare the predictions with the
receiving water biological impact. In addition, ambient tests were done in
conjunction with salinity tolerance tests to compare the agreement between the
effluent toxicity tests and the ambient toxicity where salinity itself was not
beyond acceptable ranges. Acceptable salinity was based on the concurrent
salinity tests. A marine bacterium species was also tested in which the standard
method requires salinity adjustment of the test solution so that salinity stress is
not involved.
The main purpose for the study could not be pursued because the number of
species in the estuary study was too small to use for comparisons. However, the
effluent tests could be compared to ambient tests to see how well the effluent
toxicity test predictions agreed with measured ambient toxicity.
The ambient and effluent toxicity data for daphnids agreed at all stations. Four of
six stations were correctly predicted by the fathead effluent toxicity data but the
Microtox® data for effluent and ambient toxicity did not agree. This may have
been a result of decay of chlorine toxicity in the ambient samples. Salinity in the
ambient samples had less effect than was predicted from the salinity tolerance
tests.
Considering the confounding factors that existed, the agreement between
effluent and ambient toxicity is considered good.
XIV
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Quality Assurance
Coordination of the various studies was completed by the principal investigator
preceding and during the onsite work. A reconnaissance trip was made to the
site before the study and necessary details regarding transfer of samples,
specific sampling sites, dates of collections, and measurements to be made on
each sample were delineated? The mobile laboratory was established as the
center for resolving problems and adjusting work schedules as delays or
weather affected the completion of the study plans. The principal investigator
was responsible for all Quality Assurance-related decisions onsite.
All instruments were calibrated by the methods specif fed bythe manufacturers.
For sampling and toxicity testing, the protocols described in the referenced
published reports were followed. Where identical measurements were made in
the field and laboratory, both instruments were cross-calibrated for consistency.
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/. Introduction
One of the most difficult discharge situations occurs
where freshwater effluents are discharged into saline
water. Saltwater organisms are stressed by the
freshwater effluent and freshwater organisms are
stressed by the saline dilution water making an
accurate measurement of impact difficult . Whether
freshwater or brackish water organisms should be
used for testing usually depends on thetoxicity of the
effluent. If the effluent is highly toxic the critical
mixtures of dilution water and effluent will have
salinities approaching those of the dilution water and
brackish water species would be most appropriate. If,
on the other hand, the effluent is of low toxicity,
critical concentrations of effluent will be largely
effluent and salinities will approach those of the
effluent. In this case, freshwater organisms would be
better test species.
The main approach intended in this study was to use
freshwater test species for effluent tests and compare
the results from those tests to the impact occurring in
the estuary to see if the toxicity so measured was a
valid estimate of effect for brackish water species.
Ambient tests on freshwater species were to be used
to the extent that salinity was within the tolerance of
species. The specific tolerance of the lots of test
species was to be determined simultaneously with
the effluent and ambient tests.
Because in Microtox® testing, the test solution is
adjusted to a suitable salinity, this test seemed to
offer a "bridge" between the freshwater and brackish
water species. Therefore, Microtox® testing was
included as one of the toxicity tests.
This study site was the Back River and Patapsco River
in Maryland. One publicly owned treatment works
(POTW) was located on each river within the study
area.
This report is organized into sections corresponding
to the project tasks. Following an overview of the
study design and a summary of the description of the
site, the chapters are arranged into toxicity testing,
hydrology, and ecological surveys. An integration of
the laboratory and field studies is presented in
Chapter 10. Special research study results are
presented in Chapter 11 on effluent fractionation
testing. All methods and other support data are
included in the appendixes.
1-1
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2. Study Design
The primary emphasis of this site study was the Back
River POTW and the Back River estuary. Another
POTW located on the Patapsco River, was also tested.
Study components included 7-day Ceriodaphnia
dubia toxicity tests, 7-day larval growth tests for
fathead minnows and Microtox® using a lumines-
cent marine bacterium, Photobacteria phosphoreum.
Both effluents and ambient samples were tested. A
hydrological survey of the Patapsco, Middle, and Back
Rivers for time-of-travel of the effluent was completed
and biological sampling of the macrozooplankton,
ichthyoplankton, benthic macroinvertebrate and fish
communities was done.
Difficulties were encountered in the field which
prevented completion of all the tasks on the Patapsco
River. A series of ambient stations for toxicity tests
were established but a mechanical problem with the
boat used for sampling made river bank sampling
necessary. Further, the failure to get permission to
sample from the bank at some places resulted in very
inadequate station locations. The salinity at stations
where sampling was conducted was too high to use
freshwater organisms.
2.1 Toxicity Testing Study Design
Toxicity tests were performed on the two effluents to
measure subchronic effects on the survival and
growth of larval fathead minnows and survival and
chronic reproductive effects on Ceriodaphnia dubia
(Chapter 4). A wide range of effluent concentrations
was used so that acute mortality as well as chronic
effects could be measured. The objective of these
tests was to estimate the minimum concentration of
each effluent that would cause acute mortality or
chronic effects. In addition, a salinity test was
conducted to determine the salinity tolerance of the
test organisms.
The Microtox® test was performed on effluent and
ambient samples. The test is based on the ability of a
toxicant to reduce the luminescence of a bacterium.
In addition to the effluent tests, ambient river stations
were selected on Back River from above the discharge
downstream to the confluence with the Chesapeake
Bay. Samples were also collected in the Middle and
Patapsco Rivers. Samples collected from these sta-
tions were used to measure ambient toxicity to
Ceriodaphnia dubia, fathead minnows and Microtox®.
These tests measured the loss of toxicity from the
effluents after mixing, dilution from other inputs,
degradation, and other losses such as sorbtion. These
test results would also provide data for the prediction
of ecological impact for comparison with the biological
survey data, without having to know the effluent
concentration.
2.2 Hydrological Survey Study Design
The hydrological measurements were conducted in
the Patapsco River, Middle River, and Back River by
dye studies at the two wastewater treatment plants
(Chapters 5 and 6). By modeling downstream dilution
contours for each effluent, the exposure concentra-
tions at various stations could be established. Tide
measurements were also made for the Back River.
2.3 Biological Survey Study Design
The field surveys included a quantitative assessment
of the macrozooplankton, ichthyoplankton, benthic
macroinvertebrates, and fish communities. Plank-
tonic communities in lotic systems drift with the tides
so they do not necessarily reflect exposure at the
collection site whereas the benthic community is not
nearly as mobile. Fish being quite mobile, also may be
caught in locations where they may spend very little
time.
Because an above normal incidence of tumors had
been reported in fish from the study area, the fish
captured in the survey were examined for gross
abnormalities.
2.4 Integration of Laboratory and
Field Efforts
The intent of the study was to compare the toxicity
test predictions to biological response in the estuary.
Due to an unusually cool period of weather preceding
the site study which delayed the fish spawning, the
number of species of ichthyoplankton was so small
that only subjective comparisons could be made.
2.5 Research on Effluent Fractionation
The objective of the fractionation study was to identify
the toxic components of the effluents through frac-
2-1
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tionation, toxicity testing, and chemical analyses.
Particularly for POTW effluents as distinguished from
industrial effluents, pretreatment is often the best
way to reduce effluent toxicity thus the cause of the
toxicity is needed to use this approach. The purpose
was to develop methods for toxicity identification.
2-2
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3. Site Description
Back River is tidally influenced and empties into the
Chesapeake Bay 5.6 km north of the Patapsco River
(Figure 3-1). The Back River POTW is the principal
discharger and contributed approximately 79 percent
of the river flow during the month of March 1984. The
Back River POTW is located 10.3 km from the mouth
of the Back River and receives waste from both
industrial and residential sources. The design flow of
Back River POTW is 100 mgd. A proportion of the
effluent is shunted on demand to a nearby steel mill
(which does not discharge to Back River) for cooling
water. Therefore, discharge from the POTW to Back
River may fluctuate considerably. During the study
period of March 1 984, the discharge from the POTW
averaged between 67 and 209 mgd.
The study in Back River encompased 10.3 km and
extended from the plant to the mouth of the river.
Sampling stations were:
• Station B1—located at Sandy Point upstream of
Bread and Butter Creek about 10.3 km from the
river mouth. Water depth was 1.5 m during ebb
Figure 3-1. Study area showing the two wastewater treatment plants and the biological sampling stations in Back River, Middle
River, and Patapsco River.
Patapsco
Wastewater
Treatment
Plant
Chesapeake
Bay
Plankton Tows
O Benthic Grab
• Grab samples
for water
3-1
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tide. Sediment was gray/black silt.
Station B2—located at Norristown landfill and Cox
Point about 9 km from the mouth of the river. Depth
was 1 m during ebb tide. Sediment was black silt.
Station B3—near Deep Creek about 7.9 km from
the river mouth. Depth was 2 m during ebb tide.
Sediment was gray/black silt.
Station B4—upstream from Muddy Gut and sur-
rounded by undeveloped land. Distance from the
mouth of the river is 6.3 km. Water depth was 2 m
during ebbtide. Sediment was gray/black silt with
some detritus.
Station B5—about 17 m to the right of channel
marker N 10 (red), located approximately 3.4 km
from the mouth. Depth was 3 m during ebb tide.
Sediment was gray/black silt with some clay in the
surface layer.
Station B6—at the river mouth. Depth was 3 m
during ebb tide. Sediment was gray silt with some
sand.
Station M1—located in Middle River at the con-
fluence with Dark Head Creek. Station M1 is 6.2
km from the mouth of Middle River. Water depth
was 3 m during flood tide. Sediment was gray silt.
Station M2—at the mouth of Middle River near
channel marker R4. Water depth was 4 m during
high slack tide. Sediment was black/brown silt
with some sand and many clam shells.
Station P1 —located at the Patapsco POTW at the
end of the dock. This location is in the Patapsco
River near the entrance to Curtis Bay.
Station P2—located at the Trans Maryland Ter-
minal at the end of the dock.
Station P3—located at the terminus of Chesapeake
Avenue at the Patapsco River.
3-2
-------
4. Toxicity of Effluents and Receiving Water
Toxicity tests were conducted on three species, a
daphnid (Ceriodaphnia dub/a), fathead minnow
(Pimephales promelas), and a bacterium (Photobac-
teria phosphoreum). Testing was conducted on the
Patapsco and Back River POTW effluents and ambient
stations from Middle, Back, and Patapsco Rivers.
Where effluent concentration of the ambient test
samples are known, the data from the effluent dilution
tests and the ambient tests can be compared to see
how well the effluent dilution test would predict
toxicity occurring at the ambient stations. The ambient
test data can be compared to the biological survey
data to see how well the receiving water impact was
predicted by the toxicity tests.
Because the study area was brackish water, a salinity
test was completed on the two freshwater species to
enable the effects of brackish water on toxicity to be
estimated. Since the Microtox® test utilizes a marine
bacterium, the standard protocol requires the sample
to be adjusted for salinity, so a salinity test was not
needed. However, the addition of salinity to the
samples could possibly alter the toxicity measured.
The methods used for the three tests, as well as the
details of the sampling, handling, and statistical
analyses are given in Appendix A. Routine chemistry
data is presented in Appendix E,
4.1 Chemical and Physical Test
Conditions
The Ceriodaphnia were maintained in constant
temperature cabinets at 25 ± 1°C. The mobile lab
temperature ranged from 22-26°C, but because the
fathead minnow test chambers were distributed over
three shelf levels, the temperature varied due to air
stratification. A reconstituted water control was
located at every level and the control values were not
pooled for statistical analysis. Because of this design,
the control data for each level was used for com-
parison to the exposure concentrations for each
respective level. The bacterial tests were all done at
15°C.
Tables E-1 and E-2 contain the chemistry data for
initial pH, dissolved oxygen (DO), conductivity, and
salinity plus the final DO values for the fathead
minnow tests. The final DO values for the Cerio-
daphnia tests are contained in Table E-3. Since the
exposure concentrations were made for the Cerio-
daphnia and fathead minnows as one sample, the
initial values are the same for both species. The initial
DO values are all near saturation. Temperatures of
the effluent and ambient samples ranged from 5 to
12°C as they arrived at the mobile laboratory. After
warming to test temperature (25°C), the samples had
to be aerated to reduce super saturation. Although
the final mean DO values for the fathead minnows
are all above 5.0 mg/L, individual daily values fell as
low as 2.3 mg/L. Most of these low values occurred
on day two or day three of the test. Upon finding such
values, the volume of test solution added daily was
reduced from 2 to 1 L, which resulted in higher final
DO values. Since this study was completed, other
sites with water having a high BOD and the DO below
1.0 mg/L have been encountered. In this later study,
fathead minnows had higher average weights than in
previous studies (Mount and Norberg-King, 1 986). An
assessment of this situation had led to the conclusion
that the DO measurements taken by the oxygen probe
do not reflect the micro-environmental conditions in
which the fathead minnows are living. Fathead
minnows were observed to move to a position near
the surface of the water where, in all probability, the
oxygen concentration is much higher than that
measured by the probe. Such growth at such low
measured DO concentrations would not be expected.
Apparently, the behavioc of the fish causing them to
stay near the surface when DO is low, makes the test
nearly independent of low DO effects.
The pH values changed little from initial to final;
therefore, final pH readings were not made after the
first two days. None of the initial pH values were less
than or greater than 0.5 pH units of the culture pH
values and thus did not warrant gradual transition of
the test animals. The effluents were all fresh water,
but in the ambient samples, particularly the Patapsco
River ambient samples, salinity was high (8 ppt) and
caused stress to the test animals.
4.2 Results of Fathead Minnow
Growth Tests
No comparisons of Patapsco POTW effluent dilution
toxicity test and Patapsco River ambient station tP^t
4-1
-------
data will be made due to the high salinity values (8
ppt, Table E-1), which interfered with interpretation
of the toxicity data Samples were to be collected at
designated deep-water areas; however, due to boat
problems, the ambient stations were nearshore and
the estimated effluent concentrations were not
measured.
Tables 4-1 and 4-2 contain the fathead minnow
survival and growth data for the Patapsco and Back
Table 4-1.
Seven-Day Percent Survival of Larval Fathead
Minnows Exposed to Various Concentrations of
Effluents in Reconstituted Water, Baltimore
Harbor, Maryland
Percent Effluent (v/v)
100 30
10
1
Control
Patapsco POTW
Replicate A
Replicate B
Replicate C
Replicate D
Mean
Back River POTW
Replicate A
Replicate B
Replicate C
Replicate D
Mean
0
0
0
0
0
0
0
0
100 100
100 100
90 90
90 80
95
80
100
90
90
90
93
100
100
90
90
95
80
100
100
100
98
100
90
90
90
93
100 100
90 90
90 100
100 100
95
98
100 100
100 100
100 100
90 90
98
98
'""Significantly lower than the reconstituted-water control (P
005)
River POTWs. Both effluents were diluted with
reconstituted water, as the receiving water quality
was influenced by the tide and may contain the
discharged effluent which moves upstream and
downstream in the tidal range. Survival and growth
were different from the reconstituted-water control
in the Patapsco POTW effluent only at 100 percent. In
the Back River POTW effluent, survival was only
affected at 100 percent, but growth was significantly
reduced at 30 and 100 percent effluent. The 1 and 3
percent concentrations resulted in higher weight
values than the controls, and weight at the 3 percent
effluent was significantly higher (P < 0.05) than the
control value. The calculated Acceptable Effluent
Concentration (AEC) (geometric mean of 30 and 10
percent) is 17.3 percent. This value is subject to
substantial error because of the interval between
exposure concentrations in these tests, which fol-
lowed a logarithmic dilution series.
Tables 4-3 and 4-4 contain the fathead minnow data
for all ambient stations; the stations were compared
to the appropriate reconstituted water control (Section
4-1 discusses control exposures). In the Back River
ambient stations, only Station B1 had significantly
lower survival (P < 0.05), and only Station B2 had
significantly lower growth (P < 0.05). Significantly
higher mortality (P< 0.05) occurred at all Patapsco
ambient stations, although there was no inhibition of
growth of those that survived.
Table 4-5 shows the effect of salinity (salinity test
water was derived from high quality sea water diluted
with reconstituted fresh water) on fathead minnows.
In that salinity test, survival was significantly lower at
concentrations of 16 ppt down to 4 ppt, whereas
Table 4-2.
Mean Weight (mg) of Larval Fathead Minnows Exposed to Various Concentrations of Effluents in Reconstituted
Water, Baltimore Harbor, Maryland
Percent Effluent (v/v)
100
30
10
Control
Patapsco POTW
Replicate A
Replicate B
Replicate C
Replicate D
Weighted Mean
SE
Back River POTW
Replicate A
Replicate B
Replicate C
Replicate D
Weighted Mean
SE
0.425
0480
0472
0272
0.414
0032
0288
0328
0306
0233
0 290""
0024
0.433
0480
0.400
0388
0428
0.032
0435
0470
0361
0.350
0407
0.023
0444
0.475
0436
0.388
0.435
0032
0.475
0.573
0500
0444
0.497
0024
0.435
0517
0.378
0.365
0423
0.032
0.406
0480
0420
0.394
0424
0023
0.385
0511
0.410
0375
0418
0032
0435
0429
0.378
0.356
0399
0023
":.- nnificantly lower than the reconstituted-water control (P < 0 05)
4-2
-------
Table 4-3. Seven-Day Percent Survival of Larval Fathead
Minnows Exposed to Waters from Various
Stream Stations for Ambient Toxicity, Baltimore
Harbor, Maryland
Ambient Station
Patapsco River
Replicate A
Replicate B
Replicate C
Replicate D
Mean
Back River
Replicate A
Replicate B
Replicate C
Replicate D
P1
50
40
30
30
38""
B1
80
80
80
70
P2
50
20
40
20
33la)
B2
100
70
90
80
Stream Station
P3
20
70
10
10
28'"
B3
90
90
70
70
B4
90
80
60
90
B5
90
90
80
80
B6
100
100
90
50
Mean
Middle River
Replicate A
Replicate B
Replicate C
Replicate D
Mean
78la)
M1lb)
90
90
90
80
88""
85 80 80 85
M2
100
100
90
90
95
85
Significantly lower than the reconstituted-water control for
Back River effluent control, Table 4-1
""Results shown cover a 6-day test period due to weather
conditions
growth was significantly lower only at concentrations
of 12 and 16 ppt, and not at 8 ppt. Table E-1 shows the
average salinity of the Patapsco ambient stations was
around 8 ppt, which would suggest that the fathead
minnow mortality could have been due to salinity
levels totally. Since the average salinities at all Back
River stations and Middle River stations (Table E-1)
were 1.5 ppt or less, no adverse salinity effect should
have occurred in those samples.
Table 4-7 gives the daily 7-day mean effluent
concentrations in Back River as measured by the dye
studies (Chapter 7). Mean effluent concentrations
diminished from around 28 percent at Station B1 to
18 percent at Station B6, except for Station B4 where
the mean was higher than at any other station. For
Station B4, if the one daily high value of 74 percent is
excluded, then the mean is 29 percent, very close to
B1 and B2 values. The calculated AEC of the Back
River POTW effluent was 17 percent. The effluent
concentrations at Stations B3, B5, and B6 are only
slightly higher than the AEC so measurable effects
are unlikely and none were found. An effect was
measured at Station 82, leaving only Stations B1 and
B4 where effects were expected but not found. Given
the possible error in calculating the AEC, the aging of
the effluent after discharge and possible loss of
toxicity, and the variable daily concentrations (as
opposed to the constant exposures in the effluent
test), one should consider the agreement reasonable.
Table 4-4. Mean Weight (mg) of Larval Fathead Minnows Exposed to Waters from Various Stations for Ambient Toxicily,
Baltimore Harbor, Maryland
Ambient Station
Patapsco River
Replicate A
Replicate B
Replicate C
Replicate D
Weighted Mean
SE
Back River
Replicate A
Replicate B
Replicate C
Replicate D
Weighted Mean
SE
Middle River
Replicate A
Replicate B
Replicate C
Replicate D
Weighted Mean
SE
Station
P1
0.320
0513
0283
0.283
0.357
0.063
B1
0.394
0.350
0369
0300
0355
0026
M1lbl
0.406
0.483
0467
0400
0.440
0.034
P2 P3
0 480 0 200
0.650 0564
0.288
0325 0250
0423 0460
0067 0077
B2 B3
0291 0378
0 350 0 344
0288 0307
0306 0.236
0.306"" 0.322
0025 0025
M2
0375
0585
0472
0383
0455
0033
B4 B5 B6
0411 0478 0305
0419 0.417 0375
0358 0431 0.417
0317 0419 0520
0377 0437 0387
0025 0025 0025
a Significantly lower than the reconstituted-water control for Back River effluent control, Table 4-2
""Results shown cover a 6-day test period due to weather conditions.
4-3
-------
4.3 Results of Ceriodaphnia
Reproduction Potential Tests
Table 4-8 contains the Ceriodaphnia dubia reproduc-
tive and survival data for the Patapsco POTW effluent
and the Patapsco and Middle River ambient samples.
The range of effluent concentrations initially selected
of 1 -100 percent for the Patapsco POTW effluent was
too high, and additional test concentrations were set
up which ranged from 3 percent as a high to 0.37
percent as a low. The 0.75 percent concentration was
significantly lower (P <0 05) than the control for both
survival and reproduction.Thecalculated AEC isO.53
percent (which is the geometric mean of 0.37 and
0.75).
Ceriodaphnia died quickly in all samples from the
Patapsco River ambient stations (Table 4-8). Table
E-1 reports the salinity of these stations to be about 8
ppt, which is enough to have caused the observed
response. Reproduction and survival were normal in
the Middle River samples.
The data for the Back River POTW effluent with
cumulative survival for each day is shown in Table
4-9. Both survival and young production were signifi-
cantly lower (P < 0.05) at 100, 30, and 10 percent
concentrations, but not at 3 or 1 percent exposures.
The calculated AEC is 5.5 percent.
Table 4-10 contains the reproductive and daily
survival data for the Back River ambient stations.
Survival was significantly (P < 0.05) lower at all
stations, as was reproduction except at B6. No dilu-
tions were made of these samples but some estimate
of differences in relative toxicity can be obtained by
looking at daily survival. Based on survival, Stations
B2, B3, and B4 were most toxic; Stations B1 and B5
were similar to each other and less toxic than
Stations B2, B3, and B4; and Station B6 was least
toxic.
Reference to Table 4-11 shows that even at salinity
levels of 0.25 ppt young production would be reduced,
and at the salinities measured in these samples
16
0
0
0
0
12
0
0
0
0
8
50
30
10
30
4
Survival
80
90
70
80
2
100
100
90
90
Control
100
100
90
90
30"
80"
95
Weight
Station
B1
B2
B3
B4
B5
B6
9 Mar
09
1 0
1 1
1 1
1 2
20
95
Table 4-5. Seven-Day Mean Percent Survival and Weight (mg) of Larval Fathead Minnows for Salinity Test at Baltimore Harbor,
Maryland
Salinity Concentration (ppt)
Replicate A
Replicate B
Replicate C
Replicate D
Mean
Replicate A
Hephcate B
Replicate C
Replicate D
Weighted Mean
SE
'""Significantly lower than the reconstituted-water control (P < 0 05)
Table 4-6. Daily and Mean Salinity (ppt) at Back River Stations, Baltimore naroor, Maryland
0 380
0217
—
0317
— "" — lal 0318
0 040
0500
0483
0521
0425
0481
0023
0500
0480
0417
0411
0454
0021
0305
0345
0378
0289
0329
0021
10Mar
1 0
09
1 2
1 0
1 3
23
11 Mar
1 0
1 0
1 1
1 6
2 1
2 6
12 Mar
1 3
1.2
1 1
1 0
1 5
1 7
13 Mar
1 1
1 2
09
1 0
1 5
1 9
14 Mar
1 0
1 0
09
1 0
1 2
1 9
15 Mar
1 0
1 0
1 0
1.0
1 5
22
Mean
1 0
1 0
1 1
1 1
1 3
22
SD
0 13
009
022
022
048
030
Source, Table 6-1
4-4
-------
Table 4-7. Daily and Seven-Day Mean Effluent Concentrations (%) at Back River Stations, Baltimore Harbor, Maryland
Station
B1
B2
B3
B4
B5
B6
9 Mar
43
35
9
10
16
59
10 Mar
7
6
10
74
50
18
1 1 Mar
55
19
23
13
14
7
12 Mar
3
5
15
43
9
12
13 Mar
70
63
39
28
12
9
14 Mar
2
47
33
41
21
1 1
15 Mar
17
17
34
42
16
11
Mean
28 0
27 4
23 3
35 9
19 7
18 1
SD
27 6
21 8
12 3
26 7
13 9
183
Table 4-8. Reproduction and Survival of Ceriodaphnia dubia for the Patapsco POTW Effluent and the Patapsco and Middle
Rivers Ambient Stations, Baltimore Harbor, Maryland
Patapsco POTW (v/v)
Percent Effluent
Concentration
100
30
10
3
1
Control""
3
1.5
075
0.37
Control""
Mean Number
of Young
per Female
0""
Qla.
O'al
0""
O'al
26.8
0""
Q«a,
16.3""
275
24.0
95%
Confidence
Interval
—
—
22 8-30 7
__
13 1-19.1
24.3-30 7
21 8-263
7-Day
Percent
Survival
O'a)
0""
0""
Qla)
o'al
90
O'al
Qla)
20,a,
100
80
Ambient Samples
Patapsco River
P1
P2
P3
Middle River
M1
M2
Control""
Qla.
Q.a,
o'al
292
33.8
32.2
—
27.6-308
30 0-37 6
27 1-373
Qla.
0""
Qlal
100
90
90
""Significantly lower than the reconstituted-water control (P < 0.05).
""Reconstituted-water controls
Table 4-9. Daily Survival and Mean Young Production of Ceriodaphnia dubia in Various Dilutions of Back River POTW Effluent,
Baltimore Harbor, Maryland
Back River
Percent
Effluent
(v/v)
100
30
10
3
1
Control
Cumulative Daily
Survival (%)
1
0
30
100
100
100
100
2
0
0
100
100
100
100
3
0
0
100
100
100
100
4
0
0
0
90
100
100
5
0
0
0
90
100
100
6
0
0
0
90
100
100
7
0
0
0
90
90
100
Mean Number
of Young
per Female
Qla!
0(al
Qla)
31.9
336
34.7
95%
Confidence
Interval
—
—
—
28,2-35 7
29 0-38 2
31 4-380
Significantly lower than the reconstituted-water control (P < 0.05)
4-5
-------
(Tables 4-6 and 6-1), which were from 1.0 to 2.2 ppt,
mortality should have occurred around day 4 at 2.2
ppt and about day 6 or 7 for 1 ppt salinity. It is clear
that mortality in Stations B1 through B5 occurred too
soon to be only due to salinity, whereas at Station B6,
mortality was delayed. This suggests that in either
case, the salinity in the ambient sample was not
correlated to toxicity in the same way it was in the
salinity test
As stated above, the AEC of the Back River POTW
effluent was calculated to be 5.5 percent. Table 4-7
shows the mean effluent concentrations at each
station. Mean effluent concentrations at Stations B1,
B2, and B3 ranged from 23 to 28 percent. Table 4-9
shows'that at 30 percent effluent, survival was zero
percent at 2 days, and m the 10 percent effluent, zero
percent survival at 4 days. Based on these compari-
sons, mortality at Stations B1, B2, B3, B4, and 85
occurred about as would be expected if it was due to
effluent. The mortality at Station B6 occurred con-
siderably later than it should have for effluent (or
salinity) toxicity. Since the salinity measurement is
nonspecific, one possibility is that what was being
measured as salinity was, in fact, something else.
Another possibility is that there was negative inter-
action between effluent and salinity.
4.4 Results of the Microtox® Tests
Table 4-12 contains the toxicity data from the
Microtox® test for four days for both the Patapscoand
Back River POTW effluents. The 9 March Back River
sample was a prechlorination sample and the dra-
matic difference between its toxicity and the other
samples suggests that chlorine may have been
causing the toxicity. Because of this finding, the
toxicity of pre- and post-chlorinated effluent was
Table 4-12. EC50 Values for Microtox® Tests for the Two
POTW Effluents, Baltimore Harbor, Maryland
Effluent
Back River POTW
Patapsco POTW
Test Date
9 Mar
10Mar
11 Mar
12 Mar
9 Mar
10Mar
11 Mar
12 Mar
EC50 Value
(% Effluent)
>1 00""
5.8
1.5
1 5
1 5
23
102
24
* Sample was collected before chlormation
Table 4-10. Daily Survival and Mean Young Production of Ceriodaphnia dubia in Back River Ambient Station Water, Baltimore
Harbor, Maryland
Cumulative Daily
Survival (%)
Station
B1
B2
B3
B4
B5
B6
1
100
90
100
100
100
90
2
100
0
10
0
100
90
3
0
0
0
0
50
90
4
0
0
0
0
0
90
5
0
0
0
0
0
90
6
0
0
0
0
0
30
7
0
0
0
0
0
20'al
Mean Number 95%
rvf Yfiiinn OnnfiHpnp^
ui luuny *_,UMIIUCIIL.C
per Female Interval
0""
0""
O'al
0""
2 5
-------
checked using 24-hour acute tests with Ceriodaphnia,
and no difference was found.
Table 4-13 shows the percent light reduction for the
Back River ambient stations. These samples were not
toxic enough to measure an EC50. The mean values
for light reduction show Stations B1 and B6 least
toxic; Stations B2, B3, and B4 to be similar and most
toxic; and Station B5 to be intermediate. This
sequence is similar to the mortality pattern shown by
the Ceriodaphnia chronic tests. The mean effluent
concentrations (Table 4-7) that existed at the ambient
stations were well above the EC50values. Obviously,
the effluent was less toxic in the ambient samples
than was measured in the effluent tests. This may be
due to the decay of chlorine toxicity.
Table 4-13. 15-Minute Percent Light Reduction for 91
Percent Back River Ambient Samples, Balti-
more Harbor, Maryland
Station
B1
B2
B3
B4
B5
B6
9 Mar
16.3
25.6
24.4
20.9
15.7
14.5
Test
10 Mar
14.1
22.4
16.5
25.9
17.1
11.8
Date
11 Mar
128
17.4
256
17.4
15.1
35
12 Mar
13.7
17.8
301
247
16.4
13.2
Mean
142(1.5)
20.8 (3 9)
24 2 (5 7)
22.2(3.9)
16.1 (0.9)
108(50)
4.5 Summary of Toxicity Data
The low salinity present in the Back River did not
appear to invalidate the tests with the freshwater
species. The fathead minnows were tolerant enough
of salinity that the effects could be ignored. Given a
number of factors affecting the comparison of effluent
and ambient toxicity data, expecially variable effluent
concentrations in the ambient samples, the errors in
estimating a threshold AEC, and decay of toxicity after
discharge, the agreement appears good.
For Ceriodaphnia, although salinity should have
masked the results, it did not seem to do so. At
Stations B1 through B5 there was sufficient effluent
to explain the toxicity and certainly the effects
observed at Station B6 were not likely caused by
salinity. The effluent present in the Station B6 sample
was not as toxic as would be predicted from the
effluent dilution tests.
The effluent and ambient Microtox® data do not
agree. This could be explained by chlorine toxicity in
the effluent decaying after discharge to Back River.
However, chlorine did not seem to be the cause of
toxicity with the Ceriodaphnia.
In general, the Ceriodaphnia and fathead minnow
effluent and ambient tests agreed well. When a
useful test to measure persistence of effluent toxicity
becomes available, an even better agreement might
be reached. These data do suggest that receiving
waters with salinities within acceptable ranges and
freshwater discharges can be evaluated with fresh-
water test organisms. The effluent toxicity tests, in
this case, were reasonably reliable predictors of
ambient toxicity. For much more saline estuaries,
these freshwater organisms would not be useful.
4-7
-------
5. Hydrological Studies of Patapsco River
5.1 Dilution Analysis of the
Patapsco POTW
A water tracing dye was used to tag the effluent from
the Patapsco POTW. By scaling the dye to the plant
flow, effluent dilution can be calculated throughout
the discharge plume, and the portion of effluent in
water samples taken in the area can be estimated.
Methods utilized in the dilution analysis of the
Patapsco POTW are detailed in Appendix B. Plots of
surface dilution are shown in Figures 5-1 and 5-2.
Vertical profiles of dilution are given in Table 5-1,
with their locations shown on Figure 5-2.
5.2 Evaluation of Hydrological
Conditions of the Patapsco River
The flow regime in the Patapsco River is dominated by
a three-layer, density-driven circulation pattern which
Figure 5-1. Surface dilution contours at the Patapsco
POTW, 1103 through 1217 hours, 22 March
1984. Contours are derived from data taken on
horizontal transects of plume area at high tide.
\
/Dundalk
Marine
Terminal
\
Kilometers
00 05 10
Figure 5-2. Surface dilution contours of the Patapsco
POTW, 1238 through 1417 hours, 22 March
1984. Also shown are locations of vertical
sampling stations. Contours are derived from
data taken on horizontal transects of plume area
at ebb tide.
Patapsco
Wastewater
Treatment
Plant
was originally inferred from salinity and dye meas-
urements, but which has been confirmed recently by
long-term current measurements.
The hydrodynamic explanation for the circulation is
that surface water in the Chesapeake Bay is typically
fresher, and bottom water in the Bay is typically
saltier, than water at the same depths in the Patapsco
River. As a result, there is an inflow of surface water
from the Bay, overriding the Patapsco River surface
water and an inflow of bottom water from the Bay
underriding the Patapsco River bottom water. These
two inflows are then balanced by an outflow at
middepth. The surface layer is the thinnest of the
three layers, approximately 2 m thick. The middle
layer is typically 6-8 m thick and the bottom layer 3-5
m thick.
5-1
-------
Table 5-1 Vertical Measurements of Dilution"" at Stations Surrounding Patapsco POTW Discharge, Baltimore Harbor, March
1984
Station (time)
Depth (m)
Surface
1
2
3
4
5
6
7
8
9
10
11
1 (1135)
471
471
353
353
236
236
476
1428
—
—
—
—
1A(1149)
566
472
472
472
472
237
237
1B(1201)
472
708
354
472
283
283
1C(1211)
27
32
57
83
114
131
10(1221)
26
31
38
54
65
70
IE (1230)
26
24
32
48
57
96
144
IF (1249)
32
35
38
48
57
76
—
2(1300)
29
29
32
114
114
202
2A(1311)
26
26
29
29
41
26(1319) 3(1341) 3A(1351) 3B(1403) 3C(1417) 4(1434) 4A(1443) 48(1452)
Surface
1
2
3
4
5
6
7
8
9
10
11
48
45
45
51
—
—
202
202
218
202
202
236
357
—
1412
2825
1412
1412
1412
710
41
41
48
45
54
306
101
105
101
283
473
203
108
94
101
101
189
109
27
30
29
38
54
76
83
81
76
78
78
89
98
89
98
259
720
1443
—
Note See Figure 5-2 for station locations
""Dilution is defined as the ratio of the discharge concentration to the concentration measured in the field
For short periods of time (less than 10 days), the
three-layer circulation can be overshadowed by a
wind-driven circulation in which either the surface
layer follows the wind with a counter flow at depth or
a large wind-induced set up/down in the Bay forces
water into or out of the Patapsco River at all depths.
Periods of high freshwater runoff can also generate
the usual two-layer estuarine flow, but the effect is
limited to the upper reaches of the Patapsco River and
its branches.
Residencetimesfor Baltimore Harbor can beasshort
as 3 days during strong wind events or as long as 20
days when wind and density forcing are weak. More
typically, residence time is between 8 and 10 days
when the three-layer circulation is dominant.
Velocities in each of the three layers average between
3 and 5 cm/sec and typical outflow in the middle layer
ranges between 200 and 300 mVsec. This is a
substantial flushing rate and explains why residence
times are so much shorter than would be the case for
simple tidal and river flushing.
The outfall from the Patapsco POTW discharges at a
depth of approximately 6 m which places it in the
middle, outflowing layer. Although the initial plume is
buoyant, turbulent mixing in the near field will cause
the plume density rapidly to approach that of the
ambient water and much of the effluent will remain in
the middle layer and be transported bayward at the
above-mentioned velocities. That part of the plume
which reaches the surface layer will be initially
transported upstream until vertical mixing incorpo-
rates it into the middle layer and it is flushed out.
Without a more comprehensive and detailed study, it
is not possible to quantify the average distribution of
effluent dilution.
5-2
-------
5. Hydrological Studies of Back River
6.1 Dilution Analysis of the Back River
POTW
Water samples were collected in Back River and
Middle River during the period 8-16 March 1984.
Analysis of these samples required an estimation of
the fraction of the water sample which had passed
through the Back River POTW, and, to quantify this
estimate, the plant effluent was "tagged" with a
water tracing dye.
Two problems arose with the dye tracing technique.
First, to tag all the treated water in the river would
have required injecting the dye for a longer period of
time than was economically feasible. Second, due to
the high chlorine residuals in the plant flow, the dye
injection point had to be moved into the river down-
stream of the outfall. Methods utilized in the dilution
analysis of the Back River POTW are presented in
Appendix B.2.
To address the first problem, a one-dimensional
hydrodynamic mathematical model (Hunter, 1975)
was applied to Back River and calibrated to simulate a
longer dye release, and the measured dye dilutions
were then adjusted by the ratio of the concentrations
predicted by the simulated longer release to the
actual modeled release at the location of the water
sample.
Because of the second problem, dye distribution near
the outfall can be expected to be very different from
what it would have been had the dye been injected in
the plant. The cross-sectional averaging inherent in
the one-dimensional model will mitigatethedisparity
somewhat, but the accuracy of the results will be
poorer at locations near the source.
6.2 Hydrological Modeling of Back River
Figure 6-1 shows model predictions for dye concen-
trations at three locations in Back River (Transects 5,
8, and 11; Figure B-1) versus elapsed time referenced
to the start of integration (0100 hours, 5 March
1984).
For this computer model run, the dye injection was
started on 7 March to simulate the field study.
Agreement is quite good at the mouth and, except for
the measurements on 15 March, is reasonable at the
other locations. It is not known why the 15 March
values are so high.
The calibrated model was then run again with a
simulated dye injection beginning on 1 March to
allow the simulated dye levels in the river to more
nearly reach equilibrium levels at which all effluent
present would have been tagged. As could be
expected, the model dye concentrations are higher
(Figure 6-2) at equilibrium than the previous model
run.
To estimate what the dye concentrations in the water
samples would have been had the dye injection into
the river begun six days earlier (1 March), the ratio
dye concentrations from each of the two computer
runs was multiplied by the octanol measured concen-
trations in the samples. These ratios are a function of
location and time. These predicted dye concentration
ratios were then used to calculate the percent POTW
effluent at each of the stations during the period 9-16
March, based on the steady state model with dye
levels close to equilibrium levels (Table 6-1).
6.3 Evaluation of Hydrological
Conditions of the Back River and
Middle River
It takes about two weeks for a contaminant introduced
on a continuous basis at the head of the Back River to
reach equilibrium levels throughout the river. The
model runs also show that, when the contaminant
source is turned off upstream, the lower sections of
the river are not affected for approximately 3 days.
Because the river is so shallow, tidal elevation at the
mouth is an important factor in driving an interchange
of water between the river and the bay. Large
fluctuations over periods of a few days are capable o'f
flushing the river in a relatively short time, and
estimations of river flushing rates must be understood
in this context.
6-1
-------
c
o>
g
o
o
0>
Q
5. -i
4. -
3 3.-
2.-
Transect 5 (Model)
Transect 8 (Model)
Transect 11 (Model)
+ Transect 5 (Observed)
A Transect 8 (Observed)
n Transect 11 (Observed)
' AA/yV'^^ x/.V-"'
—i—
200
—i—
400
100.
300.
500.
600.
Elapsed Time (hours)
(Relative to 0100 hours, 5 March 1984)
Figure 6-1.
Dye concentrations in the Back River as observed and predicted by the numerical model. Dye injection started at hour
62.
c
§
o
o
-------
Table 6-1. Surface Water Quality Data for Back River and Middle River Stations from 9 March 1984 Through 16 March 1984
Date
9 Mar
10 Mar
11 Mar
12 Mar
13 Mar
14 Mar
15 Mar
Station
B1
B2
B3
B4
B5
B6
M1
B1
62
B3
B4
B5
B6
Ml
M2
61
B2
B3
B4
B5
B6
M1
M2
B1
62
B3
B4
B5
B6
M1
M2
B1
B2
B3
B4
B5
B6
M1
M2
B1
B2
63
84
65
66
M1
M2
61
B2
63
B4
65
B6
M1
M2
Temperature
Time (C)
0712
0724
0732
0740
0752
0805
1010
0917
0910
0857
0845
0830
0805
0700
0730
0913
0900
0855
0843
0830
0815
0738
0751
0844
0853
0859
0909
0917
0930
1022
1005
1147
1135
1125
1115
1105
1055
1000
1020
1030
1040
1048
1058
1108
1122
1220
1236
1334
1322
1311
1304
1247
1230
• 1155
1208
1.2
1.9
2.1
23
1.9
1.8
1.5
1.7
3.5
1.3
1.8
1.3
09
1.3
1.0
38
3.6
2.1
1.9
1.5
1 5
1.8
1 4
1 4
2 1
3.1
28
1.8
20
1.8
1.4
2.5
34
3.3
2.8
20
20
2.2
1.6
28
42
57
46
3 1
24
26
2.0
6.7
74
7.1
6.9
4.9
4.5
3.4
3.3
pH
8.2
7.8
8.1
7.9
84
9.0
77
7.9
7.1
8.1
7.4
7.9
89
8.4
75
73
7.2
80
86
8.9
84
77
77
8.1
7.9
7.3
7.2
83
86
7.8
7.8
7.3
7.1
7.0
7.0
84
8.7
7.6
77
73
7 1
69
7 1
86
8.8
77
7.9
7.4
70
69
7.0
88
90
7.7
8.1
Dissolved
Oxygen
(mg/L)
14.5
14.5
15.3
144
153
15.4
133
14.5
11 7
14.4
102
13.3
150
12.4
13.2
124
11 5
142
154
16.7
147
134
13.2
14.3
140
12.0
10.2
14.2
15.0
129
13.4
12 2
11 4
90
13.7
15 2
15.5
13 1
13 6
12 1
10.2
95
93
164
145
13.1
12.7
11 1
10 1
8.3
85
16 3
160
123
13 1
Conductivity
(//mhos)
1,205
1,373
1,584
1,557
1,730
2,780
1,950
1,457
1,219
1,618
1.459
1,837
3,080
2,590
2,680
1,390
1,380
1,550
2,230
2,800
3,510
2,250
3,160
1,749
1,616
1,588
1,360
2,100
2,310
2,370
2,490
1,549
1,595
1,315
1,464
2,070
2,620
2,370
2,360
1,406
1,454
1,268
1,350
1,733
2,650
2,360
2,870
1,483
1,350
1,412
1,424
2,110
2,960
2,400
2,770
Salinity
(ppt)
0.9
1 0
1 1
1 1
1.2
2.0
1.4
1.0
09
1.2
1 0
1 3
2.3
1.9
1 9
1 0
1.0
1 1
1.6
21
26
1.6
23
1 3
1 2
1.1
1.0
1 5
1 7
1.7
1 8
1.1
1 1
09
1.0
1.5
1 9
1.7
1 7
1 0
1 0
09
1 0
1 2
1 9
1 7
2 1
1.0
1 0
1.0
1 0
1 5
22
1 7
20
Ammonia
(mg/L)
5.51
7.50
8.17
6 10
551
1 78
009
709
8.85
6.45
7.83
5.97
1 46
0.15
0 15
8.60
8.60
629
408
243
0.47
010
0.08
589
6.29
8.25
8.77
4.90
3.93
007
0.11
10.4
11 1
9.15
709
4.60
241
007
0.10
0.61
8.25
8.77
920
551
326
0.07
0.14
551
6.85
7.47
709
452
234
005
005
Percent
Effluent'"
43.0
34.9
85
10.4
15.7
585
—
6.8
5.7
10.0
740
50.1
17 6
—
—
54.5
18.8
226
134
13.9
72
—
—
34
5.0
145
42.7
8.9
11 6
—
—
700
53.4
390
282
11.8
92
—
—
23
47.3
326
41.2
20.5
11 4
—
16.7
16.6
337
423
16.2
109
—
6-3
-------
Table 6-1. (Continued)
Date Station
16 Mar B1
B2
B3
84
B5
B6
M1
M2
Temperature
Time (C)
1310
1319
1325
1335
1355
1405
1430
1440
102
87
9.8
103
6.2
52
55
54
pH
6.8
69
6.8
68
88
89
79
8.4
Dissolved
Oxygen
(mg/L)
70
5 7
63
58
16.3
15 8
12.6
13 5
Conductivity
(fjmhos)
1,198
1,331
1,250
1,288
2,830
3,650
2,400
2,920
Salinity
(PPt)
08
09
09
09
2 1
27
1 7
2 1
Ammonia
(mg/L)
775
808
8 17
800
278
1 28
007
005
Percent
Effluent""
667
304
333
493
11 5
70
—
—
Percent effluent is based on the assumption that the dye was well mixed into the average plant flow (81 mgdfrom 6 through 1 6 March)
Values further from the source are probably more accurate
6-4
-------
7. Macrozooplankton/lchthyoplankton of Back River and Middle River
7.1 Community Structure
7.7.7 Macrozooplankton
The zooplankton communities in Back River and
Middle River were overwhelmingly dominated by the
estuarine copepod Eurytemora affinis. Most of the
specimens were large, overwintering adults, the
majority being gravid females. They constituted 99.9
percent of all taxa taken at each river during both
sampling dates (Tables 7-1 and 7-2). Eurytemora
affinis was also the dominant zooplankton species
found during a study of the tidal rivers, including
Middle River (EA 1981). The amphipod Monoculodes
edwardsi was the second most abundant taxa in Back
River and the cladoceran Daphnia was second in
abundance in Middle River.
trawl collections ranged from 2.4 to 3.4°C at the
mouth of Back River where the highest numbers of
white perch were collected during the two sampling
occasions. According to Dovel (1971), most white
perch spawning occurs between 8 and 15°C in upper
Chesapeake Bay. Yellow perch, another early spring
spawner, were collected in low numbers only in
Middle River, but not enough specimens of a mature
size were taken to indicate spawning condition.
7.2 Differences Between Stations in Key
Macrozooplankton Taxa
A total of 16 macrozooplankton taxa were collected
during the two sampling dates. The number of taxa
7.1.2 Ichthyoplankton
No ichthyoplankton (fish larvae or eggs) were taken
during the two days of sampling. Gravid white perch
were collected by trawl in Back River and Middle River
during this period. None of the specimens collected
were ripe which indicates that spawning probably
had not yet occurred. Water temperatures during the
Table 7-1. Abundance and Percent Composition of the
Macrozooplankton Community of Back River
and Middle River, 12 March 1984
Table 7-2. Abundance and Percent Composition of the
Macrozooplankton Community of Back River
and Middle River, 16 March 1984
Taxa
Back River
Eurytemora affinis
Monoculodes edwardsi
Daphnia
Chaoborus
Gammarus
Ostracoda
Neomysis americana
Hemiptera
Nematoda
Middle River
Eurytemora affinis
Daphnia
Monoculodes edwardsi
Density
(No /m3)
363.9
0.086
0.005
0.004
0.004
0001
0.001
0001
0001
7496
0061
0.038
Percent
Composition
99.97
0.024
0001
0.001
0.001
<0001
<0.001
<0001
<0001
9999
0008
0.005
Taxa
Back River
Eurytemora affinis
Monoculodes edwardsi
Ceriodaphnia
Daphnia
Gammarus
Leptocheirus plumulosus
Ostracoda
Chironomidae pupae
Diptera pupae
Chaoborus larvae
Eubosmina
Neomyusis americana
Middle River
Eurytemora affinis
Daphnia
Monoculodes edwardsi
Eubosmina
Collembola
Chaoborus
Diptera pupae
Alinyraccuma
proximoculi
Density
(No/m3)
301 6
0.038
0.009
0009
0.006
0.002
0001
0001
0.001
0001
0001
0.001
8182
0.630
0.022
0012
0.008
0.006
0004
0004
Percent
Composition
9998
0.013
0.003
0003
0002
0.001
<0.001
<0.001
<0.001
<0.001
<0001
<0.001
99.92
0.077
0003
0.001
0001
0001
<0001
<0.001
7-1
-------
per station was low, ranging from three to eight in
Back River and from five to six in Middle River (Table
7-3). Combining the number of taxa from the two
collections indicated no significant differences in
number of taxa among stations (P = 0.05) (Table F-2).
£. affinis was the only taxon taken at all stations.
Monoculodes edwardsi was taken at seven of the
eight stations sampled. The other taxa were un-
common, and occurred at low densities at one to five
stations.
Abundance per station for E. affinis ranged from a
mean density of 1 9/m3 at Station B1 near the Back
River POTW(Figure 3-1 )to1,321 /m3 at Station M1 in
Middle River (Tables F-2 and F-3). Results of a 2-way
ANOVA indicated both a significant (P = 0.0001)
station and date effect for transformed densities of £.
affinis (Table F-4). A significant interaction term
suggested some inconsistency in abundance trends
between the two collection dates. However, results of
the Tukey's multiple comparison test (Sokal and
Rohlf, 1981) showed abundances at the reference
station (M1) and the lower Back River stations (B4,
B5, and B6) to be higher than those at the upper Back
River stations (B1, B2, and B3). The densities of all
other taxa combined ranged from 0.008/m3 at
Station B4 to 0.910/m3 at Station M2. All plankton
collections were made on flood tide with the exception
of Stations M1 and M2 which were sampled at ebb
tide on 16 March. The difference in tidal collections at
Table 7-3. Composition of the Macrozooplankton Com-
munity of Back River and Middle River, 12 and
16 March 1984
Station
Taxa
Nematoda
Eubosmma
Daphnia
Ceriodaphnia
Ostracoda
E affinis
N americana
A proximoculi
L. p/umulosus
Gammarus
M edwardsi
Diptera pupae
Chironomidae
pupae
Chaoborus
Hemiptera
Collembola
Total
81 82 83
0 0
X O
XO XO XO
0
O XO XO
0
O
X XO X
583
84 85
X
0
X XO
XO XO
X
XO
XO XO
X
3 8
B6
XO
XO
O
0
0
XO
6
M1
0
XO
XO
0
0
0
6
M2
O
XO
XO
XO
0
5
the Middle River stations may have influenced the
abundance of other zooplankton taxa, but did not
affect the density of £. affinis (Figure 3-1). The
general trend in abundance in Back River was an
increase in density from upriver to downriver for £.
affinis and the other taxa (Figure 7-1). This distribu-
tion is probably the result of the salinity regime in this
area which ranged fromO.6 ppt upriver to 2.1 ppt near
the mouth of Back River (Table F-5). Eurytemora is an
estuarine copepod which is typically most abundant
between 1 and 10 ppt salinity (Cronin et al., 1962).
The distribution of £. affinis in Back River is com-
parable to the results of a study by Heinle and Flemer
(1975) on the Patuxent River. During February and
March they collected the highest density of £. affinis
adults at a salinity of 2.9-5.4 ppt, respectively, and a
much lower density to no specimens at salinities less
than 1.2 ppt. In Middle River, £. affinis was much
more abundant upriver at Station M1. The salinity
was similar at Station M1 (1.3-1.4 ppt) and Station
M2 (1.5-2.1 ppt).
Figure 7-1.
0500-
0400-
g 0.300 -
c
to
0.200 H
0 1 00 -|
Spatial trends of macrozooplankton community
parameters, March 1984.
/(0910)
Other Taxa
12 March 1984
16 March 1984
1000-
c
CD
T3
-0
<
500-
M1 M2 B1 B2 83 84 85 86
Stations
l Eurytemora affinis
\ 12 March 1984
i 16 March 1984
Note. X—12 March 1984
0—16 March 1984
—i 1 1 1 1 1 1 1
M1 M2 81 B2 83 84 85 86
Stations
7-2
-------
7.3 Evaluation of the Macrozooplankton
Community
The zooplankton communities in Back River and
Middle River (reference area) were both characterized
by low diversity (number of taxa) and dominance by
theestuarinecopepod£. affinis at all stations. Similar
values for maximum abundance occurred in both
river systems, indicating no discernable response in
the Back River community to enrichment from the
Back River POTW. The density of E. affinis in Back
River increased from upriver to downriver in response
to increasing salinity levels. The freshwater input
from the wastewater treatment plant could be
contributing to the restriction of high density popula-
tions of E. affinis to the lower reaches of Back River.
7-3
-------
8. Benthic Macroinvertebrates of Back River and Middle River
Benthic macroinvertebrates were collected on 19
March 1984 at six stations in Back River and two
stations in Middle River (reference area). The objec-
tives of the study were to determine the composition
and abundance of the benthic fauna in order to assess
the response of the community to the discharge of the
Back River POTW.
The substrate type was fairly uniform from station to
station consisting mainly of fine black or gray silt with
small amounts of detritus and occasional shell
fragments, especially in Middle River. Middle River
was characterized by similar temperature levels and
low salinity at both stations. Temperature was highest
upriver in Back River near the POTW and decreased
downriver. Salinity was lowest upriver, increasing to
levels downriver which were similar to Middle River.
8.1 Community Structure
Twenty-four taxa of benthic macroinvertebrates were
collected in Back and Middle Rivers. Se\/en taxa
comprised a cumulative 90.3 percent of the total
benthos (Table 8-1). Three oligochaete taxa consti-
tuted 56.6 percent of the fauna followed by the
pelecypod Rangia cuneata (1 2.2 percent), the amphi-
pod Leptocheirus plumulosus (10.2 percent), the
polychaete Scolecolepides viridis (7.5 percent), and
Ostracoda (3.8 percent). R. cuneata was taken only at
Station M2 but at high densities. The number of taxa
at Stations B1, B3, and B4 were significantly lower (P
= 0.05) than the expected number of taxa (F-6).
8.2 Spatial Trends in Key Taxa
The oligochaete worms were the most widespread
and abundant group, and the only group found at all
stations (Table 8-2). Immature tubificid oligochaetes
without capilliform chaetae was the most abundant
taxa, comprising 24.7 percent of the total benthos.
Most of these individuals were probably in the
Limnodrilus group, the highest percentage probably
being L. hoffmeisteri. Tubificoides heterochaetus
(19.2 percent) was the second most abundant taxa
followed by L. hoffmeisteri (12.6 percent).
The number of taxa at each station ranged from 2 at
Station B4 to 13 at Station M2 (Figure 8-1). Station
M2 near the mouth of Middle River (Figure 3-2) had
numerous specimens of the pelecypods Rangia
cuneata and Mytilopsis leucophaeta. Some pelecy-
pods were also present at Station B6 in Back River
which had the next highest number of taxa (1 2). The
presence of these species and their empty shells
provides habitat which attracts more taxa. These
locations also had the highest salinity levels (2.7 ppt
at Station M2; 3.5 ppt at Station B6) (Table F-7) of any
stations sampled, which accounted for the presence
of more estuarine taxa in these areas. Only two
oligochaete taxa were present at the least diverse
station, Station B4 in Back River. The stations upriver
of Station B4 also had few taxa (3-5) and these
communities were also dominated by oligochaete
worms.
The trends in abundance distribution of the benthos
were influenced by a few and sometimes different
dominant taxa. The communities at Stations B2
through B5 had similarly low abundance, ranging
from the lowest density of 1,304/m2 at Station B4 to
1,677/m2 at Station B5 (Table 8-1). These stations
were all dominated by oligochaetes in the Limnodrilus
group, especially L. hoffmeisteri. The highest abun-
dance was at Station B6 (5,977/m2) which had a
much different and more diverse community than the
upstream stations.
Station B6 was dominated by the estuarine oligo-
chaete J. heterochaetus (4,286/m2), and less im-
portantly by the polychaete Scolecolepides virides
(846/m2) and Ostracoda (459/m2). Station B1, near-
est to the Back River POTW, also had high abundance
(4,271 /m2) but it had a less diverse habitat, dominated
by primarily freshwater oligochaetes, L. hoffmeisteri
and L. cervix, both tolerant species common in areas
with a high degree of organic enrichment (Stimsonet
al., 1982). The two Middle River stations (M 1 and M2)
had fairly high abundance (3,741 /m2 and 4,300/m2,
respectively) and more diverse communities than
most Back River stations (except B6). Station M1 was
dominated by L. plumulosus (2,451 /m2) and Station
M2 was dominated by R. cuneata (2,967/m2).
A community loss index was calculated, based on
total number of taxa, to assess differences between a
reference station (M1) and all other stations sampled
(Table 8-3). Stations M2 and B6 were most similar to
the reference station. Station dissimilarity to the
reference station was greatest at Stations B1 and B4,
especially at Station B4, since only two taxa were
collected Since relatively few taxa were taken at
8-1
-------
Table 8-1. Abundance (IMo./m2) of Benthic Macroinvertebrates Collected from Back River and Middle River, 19 March 1984
Station
M1
M2
B1
B2
Species
Imm. Tub w/o Cap. Chaet
Tubificoides heterochaet
Limnodn/us hoffmeisteri
Rangia cuneata
Leptocheirus plumulosus
Scolecolepides vindis
Ostracoda
Limnodnlus cervix
Clmotanypus L
Mytilopsis leucophaeta
Corophium /acustre
Coelotanypus L.
Pelecypoda
Nematoda
Cyathura polita
Procladius L
Monoculodes edwardsi
Heteromastus Hliformis
Chironomidae P.
Nemertea
Rhithropanopeus harrisn
A carina
Chironomus L.
Macoma mitchtlli
Number
Indivs
0.00
129.00
0.00
0.00
2451 00
473.00
358 33
000
114.67
000
71 67
000
14.33
8600
000
2867
1433
000
000
000
0.00
000
0.00
000
Pet
Comp
0.00
345
000
000
65 52
1264
958
0.00
3.07
000
1.92
0.00
038
2.30
000
0.77
038
000
000
000
0.00
000
0.00
000
Number
Indivs
2867
186.33
000
2967.00
000
47300
2867
0.00
10033
25800
000
0.00
000
2867
100.33
4300
5733
000
000
1433
14.33
000
0.00
0.00
Pet
Comp
067
433
000
69.00
000
11 00
067
000
233
600
000
000
000
067
233
1 00
1 33
0.00
000
033
0.33
000
0.00
000
Number
Indivs
2809 33
000
80267
000
000
000
0.00
645.00
0.00
000
1433
000
000
000
000
000
000
000
000
000
000
0.00
000
0.00
Pet
Comp
65.77
000
1879
000
000
0.00
0.00
15 10
000
000
034
000
000
0.00
000
000
000
000
000
000
000
0.00
0.00
000
Number
Indivs
60200
0.00
65933
000
0.00
000
000
129.00
000
000
0.00
000
000
000
0.00
000
0.00
0.00
2867
000
000
1433
0.00
000
Pet
Comp
42.00
0.00
4600
000
0.00
0.00
000
9.00
000
0.00
0.00
000
000
0.00
0.00
000
0.00
0.00
2.00
000
000
1.00
0.00
0.00
Station Total
3741 00
4300 00
4271.33
1433.33
B3
B4
B5
B6
Number
Indivs.
845.67
000
659.33
000
000
000
0.00
000
000
000
71 67
000
0.00
0.00
0.00
000
000
000
000
000
0.00
0.00
000
000
Pet
Comp
5364
000
41.82
0.00
0.00
000
000
0.00
000
0.00
455
000
0.00
0.00
0.00
000
0.00
000
0.00
000
0.00
0.00
0.00
000
Number
Indivs.
845.67
000
458.67
0.00
000
000
000
000
000
0.00
000
000
0.00
000
0.00
000
000
000
0.00
000
0.00
000
0.00
0.00
Pet
Comp
64.84
000
3516
000
000
000
000
000
000
0.00
000
000
0.00
000
0.00
0.00
000
000
0.00
000
000
0.00
0.00
000
Number
Indivs
86000
5733
48733
0.00
1433
2867
71 67
000
14.33
0.00
000
14333
000
0.00
000
000
000
000
000
000
000
000
000
000
Pet
Comp
51 28
342
29.06
000
085
1.71
427
000
085
0.00
000
855
0.00
000
0.00
000
000
0.00
000
000
0.00
000
0.00
000
Number
Indivs
14.33
4285 67
000
000
14.33
845 67
45867
000
143 33
000
000
000
11467
000
000
28.67
14.33
28.67
0.00
000
0.00
0.00
1433
1433
Pet
Comp
024
71 70
000
000
024
14 15
7 67
000
240
0.00
0.00
000
1 92
000
000
048
024
048
000
0.00
000
000
024
024
Number
Total
75071
58229
38342
370.88
30996
227.54
114.67
9675
4658
3225
1971
17.92
16 13
1433
12.54
1254
1075
358
3.58
1 79
1.79
1 79
1 79
1 79
Pet
Comp
2473
19.19
12 63
12.22
10.21
750
378
3 19
1 53
1.06
065
0.59
0.53
0.47
0.41
041
035
0 12
0 12
006
0.06
006
006
006
1576.67
1304.33
167700
5977 00
3035 88
8-2
-------
Table 8-2.
Composition of Benthic Community of Back
River and Middle River, 19 March 1984
Figure 8-1.
Spatial trends of benthic community param-
eters.
Station
Species
M1 M2 B1 B2 B3 B4 B5 B6
Nemertea X
Nematoda X X
Limnodrilus cervix X X
Limnodrilus hoffmeisteri X
Imm. tub. w/o cap
chaetae X X
Tubificoides
heterochaetus X X
Heteromastus filiformis
Scoleco/epides viridis X X
Ostracoda X X
Cyathura polita X
Leptocheirus plumulosus X
Corophium lacustre X X
Monoculodes edwardsi X X
Rhithropanopeus harnsii X
Acarina
Chironomidae pupae
Procladius larvae X X
Clinotanypus larvae X X
Coelotanypus larvae
Chironomus larvae
Pelecypoda X
Mytilopsis leucophaeta X
Rangia cuneata X
Macoma mitchilli
X
X
X
X
X
X
X
X
X
X
X
X
X
X
Total number of taxa
10 13
8 12
even the reference station, a difference of one or two
taxa made a dramatic difference in the index values.
These small differences in numbers of taxa probably
reflect patchiness in these communities which were
responsible for the wide range of values.
An index of diversity based on information theory was
calculated to examine the community at each station
(Table 8-3). In comparison with the community loss
index which considers only the number of species,
the diversity index considers the way individuals are
distributed among species. Overall, diversity was low
at all stations due to the lack of abundance of many
taxa and dominance of a few taxa at most stations.
Generally, diversity was greatest in Middle River at
Stations M1 (1.7725) and M2 (1.7614) (the reference
stations), and Station B5 (1.8942) in Back River,
which supports the trends indicated by the other data
analyses. Station B6, which had the highest number
of taxa, had relatively low diversity as indicated by the
index (1.4443) due to the numerical dominance of 7".
heterochaetus. Stations B1 through 84 had low
diversity and were dominated by oligochaetes.
5-
01
I 4
c
3
£
£
o
o
3-
2-
1-
Community Loss Index
Diversity Index
6000-
"E5000-
o
-4000
0)
-§ 3000-
c
D
< 2000-
1000-
—i 1 1 1 1 1 1 1—
Ml M2 81 B2 B3 B4 85 B6
Stations
— Abundance
' Number of Taxa
-2
o
<
ts
-12
I
8 »
-6
-4
-2
Ml M2 B1 B2 B3 B4 B5 B6
Stations
respect to abundance, number of taxa, and diversity.
These stations were most similar to the stations (B5
and 86) at the downriver portion of Back River. Much
of these similarities may be attributable to the similar
salinity regime in these areas. The community in the
upriver portion of the Back River was much different,
being characterized by low numbers of taxa and
dominance by one group, the oligochaete worms.
This was especially evident at Stations B1 and B2
immediately up and downriver, respectively, of the
Back River POTW effluent where the oligochaetes L.
hoffmeisteri and L. cervix were the dominant fauna.
These species are often the dominant organisms in
degraded freshwater and oligohaline environments.
8.3 Evaluation of the Benthos
Community
The benthic communities at the reference stations in
Middle River were fairly similar to each other in
8-3
-------
Table 8-3. Shannon-Wiener Diversity Indices (d). Associated Evenness and Redundance Values, and Community Loss Index
Calculated on Benthic Data from Back River and Middle River, 19 March 1984
Statioi.
B1
B2
B3
B4
B5
B6
M1
M2
Diversity131
1 2902
1 5330
1 2107
09355
1.8942
1 4443
1 7725
1 7614
Evenness""
0.6451
06602
0.7639
09355
06314
04029
05336
04760
Redundance""
0.355
0.3416
0.2370
00647
0.3710
05987
04681
05260
Number of
Species
4
5
3
2
8
12
10
13
Number of
Individuals
12,814
4,300
4,730
3,913
5,031
17,931
11,233
1 2,900
Community
Loss
Index'01
22500
2 0000
3.0000
5.0000
0 6250
0.1667
--
0.2308
Calculated on a log base 2
""Sum of evenness and redundance pairs is equal to 1
lclCalculated using Station 1 as reference station, (Courtemanch 1983)
8-4
-------
9. Fish Community Survey
9.1 Community Structure
The fish community of Back River differed from that in
the Middle River reference area, although water
quality characteristics measured were comparable
between areas for two sampling dates (Tables 9-1
and 9-2). In Back River on both sampling dates brown
bullhead predominated in catches and was distinctly
more abundant at Station B4 near the middle of the
river. Toward the mouth of Back River, white perch
increased in abundance as brown bullhead numbers
declined, resulting in somewhat larger total catches
downstream compared to upstream stations. In
contrast, Middle River catches were dominated by
pumkinseed, particularly at the upstream station.
White perch were also collected in Middle River, but
unlike catches in Back River, were most abundant
upstream. The number of taxa was low at all stations
and differences (P < 0.05) were not determined
among stations (Table F-8).
Back River and Middle River fish catches also differed
in the variety of species present and in the number of
fish collected per station. Trends in these parameters
are shown in Figure 9-1 in which station data are
scaled spatially by distance from the mouth of each
river. Although relatively few species were collected
in either river, slightly more were collected in the
Middle River reference area on a per-trawl basis. The
disparity was greatest on 7 March when six species
were collected at Station M1 comparedtoa maximum
of three at each of two stations in Back River. When
station catches are combined by date, the disparity
remains; seven and six species were collected at
Stations M1 and M2, respectively, compared to 3, 2,
1, 4, 3, and 4 species at Stations B1 through B6,
respectively.
The trends in total catch-per-trawl were strikingly
similar to the two sampling dates (Figure 9-1) which
lends confidence to the observed patterns. The largest
catch at any station was made at Station B4 in Back
River. Excluding these very large catches, opposing
trends in abundance are evident in the two rivers;
catches increased toward the mouth of Back River but
increased toward the headwaters of Middle River.
However, the average catch size in Back River and
Middle River was virtually identical: 53 and 55 fish
per tow, respectively, on 7 March and 64 and 60 fish
per tow on 14 March.
9.2 Fish Condition
Twenty-seven types of anomalous conditions were
observed among all fish examined from Back River
and Middle River (Tables 9-3 and 9-4). Most abnor-
malities were derived from examination of the
external surface of specimens. The variety of abnor-
malities observed per species was a function of the
number of specimens examined grossly, and no
single species appeared to display an unusually high
variety of abnormalities.
As described in the previous section, the fish com-
munities of Back River and Middle River were largely
comprised of different species, which limits inter-
area comparison of the incidence of anomalies.
Brown bullhead catfish were collected almost ex-
clusively in Back River, while pumpkinseed sunfish
were largely restricted to Middle River. Only white
perch were relatively abundant in each river.
Fifteen different conditions of abnormalities observed
among brown bullheads in Back River on the two
survey dates were recorded. Hemorrhaging of fins
andthe lower jawarea also was observed on virtually
all specimens, apparently more severely among older
fish and those collected upstream in Back River
(Tables F-9 and F-10). Although this condition was
the most obvious abnormality recorded, its ubiquitous
occurrence precluded a meaningful percent occur-
rence tally. In addition, hemorrhaging was suspected
to have been induced by the trauma of collection by
trawling; the use of set nets would be more appropri-
ate for an investigation of this abnormality.
Trends in the incidence of abnormalities among
brown bullhead in Back River are difficult to discern.
Only a few conditions were recorded for more than
one specimen or at more than one station. Therefore,
to enhance upstream/downstream differences, the
data were combined for Stations B1, B2, and B3 and
for Stations B4, B5, and B6. Fin erosion occurred
most frequently and displayed a consistent trend on
the two survey dates. It was most prevalent among
specimens collected upstream, and specifically at
Stations B2 and B3. Anotherfin anomaly, regenerated
rays, was observed six times over the two dates and
only among upstream specimens. Other conditions
observed less frequently on both dates but which
showed a higher incidence upstream include healed/
9-1
-------
Table 9-1 . Fish Catch and Water Quality Parameters, in Back River and Middle River, 7 March 1 984
Species
Brown bullhead catfish
Gizzard shad
Spotfin shiner
While perch
Channel catfish
Pumpkinseed sunfish
Yellow perch
Number of fish
Number of species
Water Quality
Depth (m)
Temperature (C)
Dissolved oxgen
(mg/L@25°C)
Conductivity (//mhos/cm)
pH
Hour
B1
6
1
1
8
3
B1
1.0
5.1
94
1,316
7.3
0925
B2
12
1
13
2
B2
20
5.0
108
1,546
75
1015
B3
17
17
1
B3
1.5
5.6
80
1,388
72
1100
Station
B4
126
1
127
2
Station
B4
20
44
15 9
2,070 2
86
1134
B5
69
10
79
2
B5
20
40
178
,520
9.0
1242
B6
1
74
1
76
3
B6
30
34
131
4,350
8.0
1418
M1
1
1
1
27
57
3
90
6
M1
2.5
4.6
12 1
2,920
78
1603
M2
2
2
7
5
3
19
5
M2
30
33
12.8
2,160
76
1712
Table 9-2. Fish Catch and Water Quality Parameters in Back River and Middle River, 14 March 1984
Station
Species
Brown bullhead catfish
Pumpkinseed sunfish
Threespine stickleback
Channel catfish
Yellow perch
White perch
Blueback herring
Number of fish
Number of species
Water Quality
Depth (m)
Temperature (C)
Dissolved oxgen
(mg/L@25°C)
Conductivity (/umhos/cm)
pH
B1
39
39
1
B1
1.0
5.6
11.2
1,693
69
B2
2
2
1
B2
1.0
63
100
1,520
7.3
B3
25
25
1
B3
1.5
63
90
1,380
76
B4
179
1
1
181
3
Station
B4
20
4.7
156
1,583 2,
82
B5
39
1
40
2
B5
2.0
3.4
157
,570
82
B6
3
1
91
95
3
B6
2.5
24
158
3,220
8.4
M1
89
8
5
10
112
4
M1
2.2
24
12 5
2,670
72
M2
4
3
1
8
3
M2
3.0
1.9
14 1
2,840
7.4
Hour
1621
1558
1515
1427
1312
1216
1002
1117
9-2
-------
Figure 9-1. Spatial comparison of fish catches in Back
River and Middle River on two days in March
1984.
o—o Middle River
-------
Table 9-3. Observations of Abnormalities by
Brown
Observation Bullhead
Body
Muscular atrophy
Healed/healing scars
Nodule/tumor
Spinal curvature (lordosis)
Unusual coloration
Small whitish spots
Small dark spots
Lesions
Fungus — smooth, opaque slime
Fins
Erosion/fin rot
Hemorrhages
(reddened membranes)
Regenerated fins, rays
Missing fin
Gills
Filament erosion
Arch cysts
Filament cysts
Gill raker erosion
Gill filament spots
Eyes
Blind
Parasites
Ergasilus
Leech
Number examined grossly
Total observation types
X
X
X
X
X
X
X
X
X
X
X
X
X
X
234
14
Species in Back River and Middle River, 7 March 1 984.
White Gizzard Yellow Spotfm Channel
Perch Pumpkinseed Shad Perch Shiner Catfish
X
X
X X
X
X X
X
X
X
X
X
XX X
X X
118 43 6 6 2 1
10 3 1201
Table 9-4. Observations of Abnormalities by Species in Back River and Middle River, 14 March 1984.
Observation
Brown
Bullhead
White
Perch
Pumpkinseed
Three-
Channel Yellow Spmned Blueback
Catfish Perch Stickleback Herring
Body
Muscular atrophy
Healed/healing scars X
Nodule/tumor X
Fungus—smooth, opaque slime
Deformed jaw
Pughead
Fins
Erosion/fin rot X
Hemorrhages
(reddened membranes) X
Regenerated fins, rays X
White cysts X
Black cysts X
Gills
Filament erosion
Gill raker erosion
Pale gill filament
Eyes
Blind X
Parasites
Ergasilus
Leech
Lernea
Number examined grossly 153
Total observation lypes 8
X
X
45
5
X
X
X
X
X
X
X
X
53
10
X
X
9
4
X
X
9-4
-------
The lower diversity of species in Back River and
dominance by brown bullhead suggest that this
species is more abundant in an environment that is
not generally favorable to survival of the endemic
fauna. Brown bullheads are described as pollution-
tolerant and omnivorous by Scott and Grossman
(1973), characteristics which allow survival under
stressful water quality conditions and adaptation to
varying types of food items. Because the basic water
quality variables measured in this study were in the
normal range, it is possible that another variable, or
perhaps food quality, accounts for the finding that
white perch were only collected at the mouth of Back
River. Although the Back River fish community
reflects a degraded environment, the average number
of fish caught per trawl was similar to that of Middle
River. This suggests that these rivers may have been
equally productive during the study period though the
quality of the catch obviously differed.
With regard to the condition of fish in Back River and
Middle River, the most consistent trend was a higher
incidence of fin abnormalities (erosion and regener-
ated rays) among brown bullheads in upper Back
River and compared to specimens collected farther
downstream. The lack of bullheads in the Middle
River reference area, however, did not allow a
determination of whether a similar upstream/down-
stream trend existed in an unpolluted area. Although
not strictly comparable, it was noted that similar fin
abnormalities occurred frequently among pumpkin-
seeds collected upstream in Middle River. There is
reason, therefore, to question whether the incidence
of fin erosion (possibly due to a bacterium; myxo-
bacterium [Post 1977]) is related to the Back River
sewage treatment plant outfall.
Robertson and May (undated report) reported that
brown bullheads collected from Back River in June
1982 exhibited branchiitis, an inflammation of the gill
epithelium. This condition increased in severity with
the proximity of specimens to the sewage outfall. In
another study, the authors found that branchiitis was
induced in white perch by exposure to chlorinated or
unchlorinated sewage effluent, again with the sever-
ity related to the effluent concentration. This trend in
anomalies could not be substantiated in the present
study, because of the methods employed, but the
suggestion of a relationship between sewage effluent
chemicals and fish condition may be related to our
finding of an absence of macroparasites on bullheads.
Brown bullheads might be unsuitable hosts for
Ergasilus and leeches, but the finding of a reduced
incidence of these parasites on Back River pumpkin-
seed and white perch compared to Middle River
specimens suggests that the sewage constituents
which induce branchiitis may be toxic to external
parasites. Such a finding would complicate the use of
parasite loading as an indicator of fish habitat quality
in Back River.
9-5
-------
10. Comparison of Laboratory Toxicity Data and Receiving Water Biological Impact
Biological field data were collected only in the Back
River outfall area. Based on the fathead minnow data,
impact would be predicted at Stations B1, B2, B3, and
B4. From the Ceriodaphnia data, impact would be
expected at all six stations. The data from Microtox®
effluent tests predict impact at all stations.
The number of species collected was entirely too few
to confidently compare test data and impact. Among
the macrozooplankton, one species comprised more
than 99 percent of all individuals, and other species
were at such low numbers that comparisons are
unduly influenced by 1 or 2 species. For the benthos,
4, 5, 3, and 2 species were collected at Stations B1
to B4, respectively, and 8 and 12 species were
collected at Stations B5 and B6, respectively, but only
1 of those was collected at Stations B1 through B4
(probably a salinity-related event). For fish, a maxi-
mum of three species was collected at a station. The
unseasonably cool weather, the salinity gradient and
the uncertain water quality of all Back River stations
makes the causes of so few species very uncertain. If
one ignores the small numbers, the trend displayed
by number of species and the toxicity are very similar,
i.e., B6 and B5 are less impacted than the rest and B1
seems to be somewhat less affected than Stations
B2, B3, and B4 (Tables 4-10, 7-3, 8-3, and 9-1).
Therefore, the comparison of toxicity data and field
impact as has been done in other reports in this series
will not be made. The daphnid, Microtox®, and
fathead effluent toxicity over-estimated ambient
toxicity at some of the stations.
70-7
-------
//. Effluent Fractionation Testing
Complex effluents are usually mixtures of dissolved
and suspended organic and inorganic components. It
is not cost-effective to chemically identify and lexi-
cologically evaluate each individual component of a
complex effluent. Chemical fractionation procedures
(Parkhurst et al. 1979; Walsh and Garnas 1983) are
useful in dividing complex aqueous effluents into
simpler subfractions, which can then be individually
screened for biological activity (i.e., toxicity) to
determine if chemical identification of a subfraction's
constituents is warranted. The purpose of this
fractionation study was to identify the primary toxic
components of complex effluents through chemical
fractionation, acute toxicity testing, and chemical
analyses.
The approach was to
• determine the relative toxicity of each subfraction
of the whole effluent and
• establish which subfraction exhibits the highest
degree of toxicity and attempt to identify chemically
the toxic constituents.
11.1 Fractionation Test Results
11.1.1 Ceriodaphnia 48-Hour Acute Tests
The acute Ceriodaphnia dubia tests on whole effluent
from the Back River and Patapsco POTWs produced
relatively similar results for the four samples tested.
The LC50 values (Table 11-1, and Figure 11 -1) for the
3-day composite and the 7-day composite were closer
for the Patapsco POTW samples (2.05 versus 3.58
percent) than for Back River POTW (1.20 and 14.6,
respectively).
For the Back River POTW samples, the organic
fraction of both composites was found to exhibit toxic
effects on Ceriodaphnia; the inorganic fractions were
not toxic. Upon testing of the base/ neutral and
acid/phenol subfractions with the 3-day composite
organic fraction, it was found that both subfractions
exhibited some toxicity, although there was an
absence of a concentration/effect relationship over a
range of concentrations (Table G-1). The highest
mortalities were noted in the next-to-lowest effluent
concentrations tested (3 percent effluent). Both the
base/neutral and acid/phenol organic subfractions
of the 7-day composite also exhibited toxic effects,
but the acute tests failed to elicit a concentration/
effect response over the range of concentrations
tested (Table G-1). Maximum mortalities observed
(50 percent) occurred in the 100 percent effluent
concentration for both 3- and 7-day composites, so
the LC50 values were not calculated but were
estimated to be approximately 100 percent.
The Patapsco POTW results were slightly more
complicated. The 3-day composite whole effluent
sample had an LC50 value of 2.1 percent, the organic
fraction had an LC50of 9.3 percent andthe inorganic
fraction had an LC50 of 37.6 percent. The base/
Table 11-1.
LC50 Values (in % Effluent) Calculated by Moving Average Method, Based on Ceriodaphnia dubia 48-Hour Acute
Tests""
Whole
Effluent
Inorganic
Fraction
Cation
Fraction
Anion
Fraction
Organic Base/Neutral Acid/Phenol
Fraction Fraction Fraction
Back River POTW
3-Day Composite Mean 1.20 Not Toxic NA
95% Confidence Limits <0.01-495
7-Day Composite Mean 146 Not toxic NA
95% Confidence Limits 7.9-313
NA
NA
548"
430
285-74.0
Not
calculated
-100
Not
calculated
-100
Patapsco POTW
3-Day Composite Mean
95% Confidence Limits
7-day Composite Mean
95% Confidence Limits
2.05
05-4.13
358
2.19-6.32
37.6
247-61 8
Not toxic
54.8lal
Not
required
Not toxic
Not
required
9 18
596-16.2
173(bl
4 16
0.97-11 2
774
1 96-225
Not toxic
80 3""
"See Figure 11-1
''Calculated by the binomial procedure
11-1
-------
Figure 1*1-1. Schematic results (LC50 in percent effluent)
of Ceriodaphnia acute tests on effluent frac-
tions.
Back River POTW
Base/Neutral
Organic
3-Day
Composite
Whoie
7-Day
Composite
Patapsco POTW
Organic
Whole
Composite ( 2-05 j
3-Day
Composite
Whole
r^Yl
v!!y
neutral fraction of the 3-day composite sample
exhibited acute toxicity to Ceriodaphnia (4.16 percent
LC50), whereas the acid/phenol fraction did not. The
inorganic fraction was further split into cation and
anion fractions The LC50 value for the cation fraction
was 54.8 percent, whereas the anion fraction did not
result in sufficient mortality to calculate an LC50
value (Table G-1). Thus, the majority of the toxicity
noted in the 3-day Patapsco POTW composite was
attributable to the base/neutral subfraction but there
was some toxicity in the cation fraction. The toxicity
response to the 7-day Patapsco POTW composite was
similar to that noted for the Back River POTW samples
in that the inorganic fraction was not toxic (Table
11-1). The organic fraction was less toxic (17.3
percent LC50) than the whole effluent (3.58 percent
LC50). The base/neutral and acid/phenol subfrac-
tions both displayed some toxicity, although the LC50
values indicate that the base/neutral subfraction
was considerably more toxic (7.7 percent LC50) than
the acid/phenol subfraction (80.3 percent LC50).
In summary, the whole-effluent toxicities of the Back
River and Patapsco POTWs were similar, but, after
fractionation, the organic fraction (which contributed
the most to the overall toxicity of the four samples
tested) of the Back River POTW effluent had con-
siderably less toxicity than the whole effluent. In
contrast, the organic fraction of the Patapsco com-
posites was nearly as toxic as the whole effluent, and
most of the toxicity of this fraction was traceable to
the base/neutral subfraction.
11.1.2 Microtox® Tests
The fractionation results of the Microtox® tesst were
different from the Ceriodaphnia tests. The whole
effluent, which exhibited the second greatest toxicity
to Ceriodaphnia (Patapsco POTW 3-day composite),
was the least toxic according to the Microtox® tests
(Table 11-2 and Figure 11-2). Conversely, the Back
River POTW 7-day composite whole effluent, which
displayed the geatest toxicity according to the Micro-
tox® tests, was the least toxic according to the
Ceriodaphnia tests.
Only the Back River POTW whole effluent samples
displayed toxicity in the Microtox® tests. The 7-day
composite was the more toxic of the two effluent
samples from Back River POTW (3.0 percent EC50
value compared to 28 percent for the 3-day com-
posite). Neiter the organic nor inorganic fraction of
the 3-day composite proved toxic according to Micro-
tox® EC50s. The 7-day organic fraction was slightly
toxic, with an EC50 value of 38.7 percent effluent.
Samples with Microtox® EC50 values greater than
45.5 percent were classified as nontoxic because
those values must be extrapolated. Extrapolated
values (Table 11 -2 and Figure 11 -2) are provided only
as a rough indication of toxicity. Because the organic
fraction displayed limited toxicity, and since the
Microtox® instrument was temporarily inaccessible
when the organic samples were processed, the
base/neutral and acid/phenol subfractions were not
tested for Microtox® toxicity. The inorganic subfrac-
tion was not toxic according to Microtox® EC50
values. The Microtox®EC50 results agreed with the
acute Ceriodaphnia tests in suggesting that the
inorganic fractions of the Back River POTW effluent
were not toxic.
11-2
-------
Table 11 -2. EC50 Values (in percent Effluent) Based on Beckman Microtox® Acute Tests"
Whole
Effluent
Inorganic
Fraction
Cation
Fraction
Anion
Fraction
Organic
Fraction
Base/Neutral Acid/Phenol
Fraction Fraction
Back River POTW
3-Day Composite
7-Day Composite
Patapsco POTW
3-Day Composite
28.0
3.0
Not toxic
(-100)
Not toxic
Not toxic
Not toxic
O45.5)
NA
NA
Not toxic
(781)
NA
NA
Not toxic
(46.6)
Not toxic
(51.5)""
387
Not toxic
(61.8)
NA
NA
la'See Figure 11-2.
""Any Microtox® EC50 >45 5 percent is extrapolated and is considered not toxic.
NA
Not tested Not tested
NA
7-Day Composite
Not toxic
(48)
Not toxic
(95.5)
NA
NA
Not toxic
(66.3)
NA
NA
Figure 11-2. Schematic results (EC50 value in percent
effluent) of Microtox® tests on eflfuent frac-
tions.
Back River POTW
3-Day
Composite
Organic
7-Day
Composite
Patapsco POTW
3-Day
Composite
Organic
Not
Toxic
-(61 8)J
Inorganic
Cation
7-Day
Composite
Whole
'Not"
Toxic
J48)
Inorganic
Not
Toxic
L(955)J
The Patapsco POTW effluent, both 3-day and 7-day
composites, were found not toxic in the Microtox®
tests, in contrast to their toxicity to Ceriodaphnia. The
inorganic and organic fractions were tested by
Microtox® for both composites, and were found to be
not toxic (EC50 values >45.5 percent). Because the
cation and anion subfractions of the 3-day composite
had been tested using the Ceriodaphnia 48-hour
acute test, their toxicities were evaluated by Micro-
tox® as well. Both subfractions proved not toxic (EC50
values >45.5 percent).
11.1.3 Chemical Analyses of Toxic Fractions
The base/neutral subfractions of the organicfraction
of the 3-day and 7-day Patapsco POTW effluents
were selected for chemical analyses due to the
toxicity observed in the Ceriodaphnia acute tests.
These subfractions were analyzed for pesticides,
herbicides and PCBs by gas chromatography, and for
base/neutral priority pollutants by gas chromatog-
raphy/mass spectrometry (GC/MS) (Appendix G).
Levels of pesticides, herbicides, and PCBs (Table 11 -
3) and base/neutral priority pollutants (Table 11-4)
were below detection limits for both the 3-day and
7-day composite Patapsco POTW samples.
Results of the GC/MS analyses for base/neutral
organic compounds, including reconstructed ion
chromatograms and quantitation reports for samples,
standards, spikes, and blanks, are included in
Appendix G.
11.2 Summary
The organic fraction contributed the most to the
overall toxicity of the four effluent samples tested.
However, the toxicity of a particular waste was not
always traceable to one particular subfraction (i.e.,
base/neutral or acid/phenol). For the Patapsco
POTW, the base/neutral subfraction accounted for
the majority of the observed toxicity. Chemical
11-3
-------
analyses on the base/neutral subfractions did not
identify the toxic components among the pesticides,
herbicides, PCBs, and priority pollutants tested.
Toxicity, as measured by the acute Ceriodaphnia
tests, were different than the toxicity as measured by
the Microtox® test.
Table 11-3. Levels of Pesticides, Herbicides, and PCBs in
3-Day and 7-Day Composite Patapsco POTW
Effluents
Concentration (yug/L)
Compounds
Aldrin
alpha BHC
beta BHC
delta BHC
Linda ne
Chlordane
p,p'-DDE
p,p'-DDD
p,p'-DDT
Dieldrin
Endosulfan 1
Endosulfan 2
Endosulfan sulfate
Endrm
Endrin aldehyde
Heptachlor
Heptachlor epoxide
Methoxychlor
Mi rex
Toxaphene
Aroclor 1016
Aroclor 1221
Aroclor 1 232
Aroclor 1 242
Aroclor 1 248
Aroclor 1254
Aroclor 1 260
2,4-D
2,4,5-TP
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
Less than
3-Day
0001
0.0006
0006
0.001
00007
002
0.002
0005
0.007
0.003
0.003
0004
0.008
0.008
0.009
0003
0002
0.02
0.009
0.3
0.03
0.1
0.04
0.03
003
004
0.05
0.02
0.003
7-Day
0.001
0.0005
0.005
0.0009
00006
0.02
0.002
0.005
0.007
0002
0002
0.004
0.007
0007
0.008
0.003
0001
001
0.008
03
003
0.1
004
0.03
003
004
005
001
0003
Isophorone
Bis(2-chloroethoxy)methane
1 ,2,4-Tnchlorobenzene
Naphthalene
Hexachlorobutadiene
Hexachlorocyclopentadiene
2-Chloronaphthalene
Acenaphthylene
Dimethyl phthalate
2 6-Dmitrotoluene
Acenaphthene
2 4-Dmitrotoluene
Fluorene
Diethyl phthalate
4-Chlorophenyl phenyl ether
N-Nitrosodiphenylamme
1 2-Diphenylhydrazme
4-Bromophenyl phenyl ether
Hexachlorobenzene
Phenanthrene
Anthracene
Di n-butyl phthalate
Fluoranthene ...
Benzidine
Pyrene ....
Butyl benzyl phthalate
Benzo(a)anthracene
3 3'-Dichlorobenzidine
Chrysene
Bis(2-ethylhexyl) phthalate
Di-n-octyl phthalate ....
Benzo(a}pyrene .....
lndeno{1 2 3-cd)pyrene
Dibenzo(a,h)anthracene
Unresolved Isomenc Pairs
Benzo(b)fluoranthene+
benzo(k)fluora nthene
Less than
. . Less than
. . Less than
Less than
Less than
Less than
Less than
Less than
. . Less than
. . Less than
. . Less than
. . Less than
. . Less than
Less than
Less than
. . Less than
Less than
Less than
Less than
. . Less than
. . Less than
Less than
Less than
Less than
. . Less than
. . Less than
. . Less than
. . Less than
. . Less than
. . Less than
Less than
. . . Less than
. , Less than
, . . Less than
Less than
. . . . Less than
0.40
040
040
040
0.40
1.2
0.40
040
0.40
0.40
0.40
0.40
040
0.40
0.40
040
0.40
0.40
1 2
040
0.40
0.40
0.40
80
0.40
0.40
040
1 2
0.40
4.0
0.40
0.40
0.80
0.80
080
0.80
037
0.37
037
037
0.37
1 1
037
037
0.37
0.37
0.37
037
0.37
037
0.37
037
0.37
037
1 1
037
0.37
0.37
0.37
7.5
0.37
0.37
037
1 1
037
37
0.37
037
075
075
0.75
075
Table 11-4. Levels of Base/Neutral Compounds, Deter-
mined by GC/MS Analysis (EPA Method 625),
for 3-Day and 7-Day Patapsco POTW Effluents
Base/Neutral Compounds
3-day 7-Day
N-Nitrosodimethylamme Less than 12 1.1
Bis(2-chloroethyl)ether Less than 0.40 0.37
1,3-Dichlorobenzene Less than 0.40 037
1,4-Dichlorobenzene Less than 0.40 037
1,2-Dichlorobenzene Less than 0.40 037
Bis(2-chloroisopropyl)ether Less than 0 40 0 37
Hexachloroethane Less than 040 037
N-Nitroso-di-n-propylamme Less than 080 0.75
Nitrobenzene Less than 1.2 11
11-4
-------
References
American Public Health Association, American Water
Works Association, and Water Pollution Control
Federation. 1980. Standard Methods for the Exam-
ination of Water and Wastewater, 15th edition.
APHA, Washington, 1,134pp.
Beckman Microtox® System Operating Manual.
1982. Beckman Instructions 015-555879. Beck-
man Instruments, Inc. Microbics Operations,
Carlsbad, CA 92008.
Courtemanch, D. L. 1983. The use of a coefficient of
community loss to assess environmental degrada-
tion. Presented at the 31st Annual Meeting, North
American Benthological Society, 27-29 April, 1983,
LaCrosse, Wl.
Cronin, L. E., J. C. Daiber, and E. M. Hulbert. 1962.
Quantitative seasonal aspects of zooplankton in the
Delaware River Estuary. Chesapeake Sci., 3:63-93.
Dovel, W. L. 1971. Fish eggs and larvae of the upper
Chesapeake Bay. NRI Spec. Rept. No. 4. Natural
Resources Institute, Univ. Md. 71 pp.
Ecological Analysts, Inc. (EA). 1974. Perryman Site:
Power Plant Site Evaluation, Aquatic Biology, Final
Report. PPSE 2-2. Prepared for Maryland Power
Plant Siting Program, Annapolis, MD. 193 pp.
Ecological Analysts, Inc. (EA). 1980. C. P. Crane
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Ecological Survey of the Aquatic Biota. Annual
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Ecological Analysts, Inc. (EA). 1981. C. P. Crane
Power Plant: An Environmental Assessment and
Ecological Survey of the Aquatic Biota, Final Report,
1978-1980. Prepared for Baltimore Gas and Electric
Company. EA Report BGE02K2.
Hamilton, M. A. 1984. Statistical Analysis of the
Seven-Day Ceriodaphnia reticulata Reproductivity
Toxicity Test. EPA Contract J3905NASX-1. 16
January. 48 pp.
Heinle, D. R. and D. A. Flemer. 1975. Carbon
requirements of a population of the estuarine
copepod Eurytemora affinis. Marine Biology,
31:235-247.
Hunter, J. R. 1975. A one-dimensional dynamic and
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Marking, L. L. and V. K. Dawson. 1973. Toxicity of
QuinaldineSulfatetoFish. Invest. Fish. Contrib. No.
48. U.S. Fish and Wildlife Service, Washington, DC.
8pp.
Mount, D. I. and T. J. Norberg. 1984. A seven-day life
cycle cladoceran toxicity test. Environ. Toxicol.
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Mount, D. I. and T. J. Norberg-King (1986). The
Validity of Effluent and Ambient Toxicity Test for
Predicting Biological Impact, Kanawha River,
Charleston, West Virginia. EPA/600/3-86/006.
Norberg, T. J. and D. I. Mount. 1985. A new fathead
minnow (P//r7ep/7a/esp/-ome/as)subchronic toxicity
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Parkhurst, B. R., C. W. Gehrs, and I. B. Rubin. 1979.
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Post, G. 1977. Glossary of Fish Health Terms. Fish
Health Sect., Amer. Fish. Soc., Washington, DC. 48
pp.
Robertson, P. G. and E. B. May. No date. Pathological
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Stimpson, K. S., D. J. Klemm, and J. K. Hiltunen.
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(Annelida: Clitellata: Oligochaeta) of North America.
EPA/600/3-82/033. April. 61 pp.
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R-1
-------
Appendix A
Toxicity Tests and Analytical Methods
A.1 Sampling and Sample Preparation
Sampling of Patapsco and Back River POTW was
done using ISCO* samplers set to collect an aliquot
every 15 minutes and to composite the sample into a
five-gallon polyethylene container. About 15 L of
sample was collected each 24-hour period and a new
composite sample was taken each day. On the first
two collection days, 9 and 10 March, unseasonably
cold weather froze the ISCO samplers and a grab
sample had to be used.
The Back River and Middle River ambient samples
were taken at low slack tide as a grab sample, at 0.5
meters in depth. The three Patapsco River ambient
samples were grab samples taken between 8:00a.m.
and 12:00 noon each day. About 1 6 L were collected
in collapsible polyethylene containers.
Reconstituted water was made using the formula of
Marking and Dawson (1973) (moderately hard option)
at the Environmental Research Laboratory in Duluth,
Minnesota, and stored in five gallon polyethylene
jugs. Water was kept at room temperature until used.
All effluents were diluted with reconstituted water.
The salinity test was set up using seawater diluted
with the same reconstituted water stock to make the
appropriate salinity test concentrations. The seawater
was provided by the EPA-Narragansett and was from
their laboratory seawater supply.
Effluent dilutions were made using polypropylene or
polyethylene laboratory ware. The values were
measured using graduated cylinders of various sizes
and 4 L beakers for mixing. Samples were warmed to
25°C and then aerated until supersaturation was
removed as measured by dissolved oxygen levels of
8.5-9.0 mg/L. For the effluent dilution tests, 100
percent effluent and 100 percent dilution water were
warmed separately and aerated before being mixed.
All samples were used within six hours of collection.
Two liters of each exposure water was made and 1 70
ml was used for the Ceriodaphnia tests and the
remainder usedforthefathead minnowtest. Because
of BOD in some samples, the daily renewal volume for
the fathead minnow test was reduced to 1 L in the
Back River ambient samples on day 4 of testing.
'ISCO, Inc , Lincoln, Nebraska
After the 2 L was prepared, DO, pH, conductivity
and/or salinity was measured. When the daily
renewal was made, DO was measured in one
compartment of each chamber in the fathead minnow
test and in one cup of the Ceriodaphnia test in each
exposure. At least once, DO was measured in the
fathead minnow tests soon after the lights were
turned on to determine diurnal DO cycles, but none
were found.
A.2 Ceriodaphnia Test Methods
The protocol followed in general that of Mount and
Norberg (1984) with a few exceptions. A hard,
transparent, plastic, one-ounce cup was used in place
of 30-ml glass beakers, and the cups were discarded
after use. Each day, a new and different sample of
effluent or ambient water was used. The initial
measurements, for pH, DO, salinity, and conductivity
were made on the 2 L volume and are pertinent for
both tests. For the final DO measurement, one cup
from each exposure condition was used to measure
final DO.
A new food formulation was used which consisted of
three parts: (1) 5 g/L of dry yeast; (2) 5 g/L of
Cerophyl®* , stirred overnight and filtered through a
plankton net; and (3) 5 g/L of trout chow, aerated
vigorously for seven days, settled and decanted. The
yeast suspension and the supernatant from the
Cerophyl® and trout chow are mixed in equal parts,
and new food was made every seven days. The
mixture was kept refrigerated as are the Cerophyl®
and yeast components, while the trout chow super-
natant remained frozen until the mixture was made.
In our experience, this food was suitable for a wide
variety of water types, including reconstituted water.
Because the suspended solids concentrations are
—1,800 mg/L, which is less than half the solids
contained in the yeast suspension, this mixture is fed
0.1 ml per day per Ceriodaphnia rather than 0.05 ml
as was recommended for yeast (Mount and Norberg
1984).
*CerophyK"), was obtained from Agri-Tech, Kansas City, Missouri As of
January 1985, Cerophyl® was no longer produced by that manufacturer
A-1
-------
All test animals were less than 2-hours-old and were
produced from adults that were 11-14 days of age.
The cultures were at pH 7.1 and no acclimation to pH
was necessary when the test animals were placed in
the exposure chambers.
A.3 Fathead Minnow Test Method
The methods for the fathead minnow test followed
closely those described by Norberg and Mount (1985).
The test chambers were 30.5 x 5.2 x 10.2 cm, and
divided into four compartments; this design allowed
four replicates for each concentration. Less than 24-
hour-old posthatch fathead minnow larvae were air
shipped from the Duluth culture to the mobile
laboratory, and were assigned to the exposure
chambers immediately upon arrival. The fish were
assignedtothetest compartments by pipetting one or
two fish at a time to each replicate test chamber until
all replicates had 10fish in each or 40 per concentra-
tion. Uneaten brine shrimp were removed daily by
siphoning the tanks during test solution renewal. At
the same time, the volume in the test chamber was
drawn down to 1 cm, after which 2 L of new test
solution was added. Because the Back River ambient
samples had a significant BOD, the volume put in
each chamber daily was reduced to 1 L on day 4 of the
test to improve the surface-to-volume ratio. A 16-
hour light photoperiod was used.
After 7 days of exposure, the fish were preserved in 4
percent formalin. Prior to weighing, they were rinsed
in distilled water. Then each group was dried for 1 8
hours in preweighed aluminum pans and weighed on
a five-place analytical balance.
A.4 Ceriodaphnia Statistical Analyses
The statistical analyses of the Ceriodaphnia data
were performed using the procedure of Hamilton
(1984) as modified by J. Rogers (1984). The essential
features of the analysis are that a mean young
production per live adult is calculated for each day
young were observed, and these means are summed
over the period of the test to give a 7-day estimated
mean production peradult, ignoring mortality(all data
method). In this way, the adults which die during the
test do not reduce the estimate of young production.
The variance and confidence intervals of the esti-
mates were derived from a distribution generated by
the bootstrap method, using a sample size of 999. The
multiple comparisons for effluents were made using
Dunnett's test. Multiple comparisons for ambient
toxicity tests are made using Tukey's Honestly
Significant Difference Test. The multiple comparison
procedures were modified to compensate for different
variances and degrees of freedom for different tests.
The survival, defined as the number of adults alive at
the beginning of the last observation period was
transformed using an arcisine transformation for
binomial proportions. The variance and confidence
intervals of the transformed survival and the corre-
lation of the survival and reproduction estimates
were derived from the bootstrap method as above.
The multiple comparisons for the survival followed
the same procedures as for the reproduction.
A.5 Fathead Minnow Statistical Analysis
The four mean group weights are statistically ana-
lyzed with the assumption.that the four compartments
behave as replicates. The method of analysis used
assumes the variability in the mean treatment
response as proportional to the number of fish per
treatment. MINITAB (copyright Pennsylvania State
University, 1982) was used to estimate a t-statistic for
comparing the mean treatment and control responses
using weighted regression with weights equal to the
number of measurements in the treatments. The t-
statistic is then compared to the critical t-statistic for
the standrd Dunnett's test (Steel and Torrie 1960).
Prior to the regression analysis, the survival data are
arcsine transformed (which is a variance-stabilizing
transformation).
A.6 Microtox® Testing Methods
The Microtox® System was utilized to conduct toxicity
tests on both the effluent and ambient samples.
Procedures for the tests followed those described in
Beckman's "Microtox System Operating Manual."
This toxicity test is based on increases or decreases in
the natural light emissions of the luminescent marine
bacteria Photobacteriumphosphoreum (Beckman no
date). All tests were performed on the Beckman
Microtox® Model 2055 Toxicity Analyzer. Turbidity
was determined not to be a problem with any sample.
The color correction method was not used on any of
the tests. The instrument was calibrated each day
according to manufacturer's specifications. All data
were recorded permanently on Beckman Microtox®
chart paper.
A.6.1 Microtox® Effluent Samples
All effluent test concentrations were prepared using
serial dilutions of 2:1 or 3:1. The salinity of all
samples was adjusted to 2 percent NaCI using
Microtox® osmotic adjusting solution prior to the
preparation of dilutions. The effluent samples were
run in dupliate using four or five concentrations and a
control. If 100 percent sample were to be tested, it
was run separately from the serial dilutions with its
own control. All 100 percent samples were treated
identical to the ambient stations; this resulted in a
final concentratoin being assayed of 90.1 percent. All
A -2
-------
dilutions were made using Microtox® diluent. The
lyophilized reagent bacteria was rehydrated using
Microtox® reconstitution solution. Ten microliters of
the reagent was then introduced into each of the 10
cuvettes to be charged with the test solutions. The
reagent was allowed to acclimate for 1 5 minutes and
at the end of this time period the light output from
each cuvette was measured. Immediately after this
initial reading (I0), each cuvette was charged with test
solution, and at the end of five minutes (I5) and 15
minutes (Ii5) the light output from each cuvette was
recorded again. All data were recorded on Beckman
Microtox® chart paper and normalized using the
Sharp Model EL1500 calculator. Toxic effects were
defined as the concentration causing 50 percent
reduction in light output after 5 or 15 minutes
exposure to the effluent (5EC5o, 15EC5o). Effect
concentrations for those effluents tested at 100
percent (90.1 percent actual concentration) were
based on extrapolations.
A.6.2 Microtox® Ambient Samples
All ambient samples were salinity adjusted to 2
percent NaCI using Microtox® osmotic adjusting
solution. This adjustment resulted in a final test
concentration of 90.1 percent. Each sample and
control was run in duplicate or triplicate depending on
the time available. The tests were initiated by
pipetting 10 >ul of rehydrated bacteria reagent into
each of the cuvettes containing sample. Five and
fifteen minutes after the introduction of the reagent,
light measurements were recorded. These data were
reduced by calculating the mean percent differences
in light output between the control and each sample
tested. These differences were interpreted as either
an increase in light output (stimulation) or a decrease
in light output (inhibition).
A -3
-------
Appendix B
Hydrological Sampling and Analytical Methods
B.I Patapsco River Survey
B. 1.1 Dye Injection
A 20 percent solution of rhodamine WT dye was
injected into the Patapsco POTW flow at the down-
stream end of the chlorine contact chamber, just
upstream of the pump. Injection began at 1345 hours
on 21 March and was terminated at 1550 hours on 22
March. During that time 37.3 Ibs of solution were
pumped, which is equivalent to 3.6 x 10~2 g/sec of
pure dye.
The average flow through the plant on 22 March was
37.9 mgd (million gallons per day), or 1.66 x 106
g/sec. Therefore, the average dye concentration at
the discharge was
-2
3.6x10
1.66x106
= 21.7ppb
(Equation B-1)
B.I. 2 Dechlorination
Chlorine residuals in the Patapsco POTW effluent are
high enough to oxidize the rhodamine molecule. To
prevent this, a 38 percent solution of sodium thio-
sulfate was injected along with the dye. The sodium
thiosulfate is acted on preferentially by the chlorine
and the rhodamine remains intact provided thiosul-
fate concentrations remain about 5.6 times the
chlorine concentrations (APHA et al. 1981, p. 786).
The injection rate of the thiosulfate was 690 ml/min,
which for a plant flow of 37.9 mgd will protect the
rhodamine against chlorine residuals up to 0.6 mg/L.
B.I .3 Dye Sampling Procedures
Dye was sampled on 22 March from two boats, one
making horizontal measurements and the other
making vertical measurements. Each boat was out-
fitted with a Turner Designs Model 10 fluorometer in
the continuous-flow configuration, a temperature
sensing device, and a sampling pump. The fluorom-
eter is capable of measuring Rhodamine dye to
concentrations of 0.01 /ug/l. Decay processes of the
Rhodamine dye were considered to be minimal, if
any. Standard fluorometric practices were used.
The boat making horizontal measurements had a rigid
airfoil-shaped probe attached to its side. Polyethylene
tubing was inserted through this probe and fed to the
fluorometer intake. From the fluorometer, the tubing
led to the temperature sensor and from there to the
sampling pump and back over the side. The end of the
probe was 0.5 m below the surface. The boat
traversed the dye plume in a "ladder" fashion
following the dye upstream and downsteam until
fluorescence levels fell to background values.
The boat making vertical measurements had a weight
affixed to the end of the sampling tubing, but was
otherwise configured the same. Measurements were
made from the surface to the bottom in 1-m incre-
ments.
The "horizontal" boat navigated using a Motorola
Mini-Ranger system. The "vertical" boat used an
electronic distance meter (EDM) with a person on
shore who would note the distance and measure the
angle between the boat and a reference direction
using a surveyor's transit.
B.2 Back River and Middle River
B.2.1 Dye Injection and Sampling Procedures
Dye was injected from an anchored dinghy approxi-
mately 50 yd downstream of the treatment plant
outfall. The dye was a 20 percent solution of
rhodamine WT and was pumped into the water at a
rate of 12 ml/min using a precision metering pump
driven by a 12 VDC automotive battery. The pump was
started at 1445 hours on 7 March 1984.
On the morning of 17 March, it was discovered that
the battery had been stolen and, since the injection
equipment had been seen to be working shortly
before 1600 hours on 16 March, it is estimated that
injection stopped around 1700 hours on 1 6 March.
Two boats were used to map the distribution of the
dye. Each was equipped with a Turner Designs Model
10 fluorometer, a temperature sensing device, and a
sampling pump. Water was drawn in through a probe
mounted to the side of the boat 0.5 m below the
surface, and was then passed through polyethylene
tubing to the fluorometer, the temperature sensor,
the sampling pump, and then back over the side. This
B-1
-------
procedure enabled a continuous record of dye-
induced fluorescence to be obtained as a boat
traversed a river transect. The temperature sensor is
necessary because dye fluorescence is a function of
temperature, and fluorometer readings must be
related to instrument calibrations through a common
temperature to which all values are corrected.
One boat sampled Transects 2A through 6 (Figure
B-1), and the second boat sampled Transects 7
through 11. Transects 1 and 2 had to be abandoned
because the water was too shallow. Mappings were
done on 11,13,15,17, and 20 March as summarized
in Table B-1. Boat position was interpolated assuming
a constant speed from bank to bank.
B.2.2 Tide Measurements
A Stevens Model F-68 recording tide gauge was
placed at the mouth of the river on the south side at
Cuckold Point. The record has several breaks due to
icing conditions in the stilling well, as well as wave
overtopping during unusually high seas. The breaks
were filled in by correlating the usable record with the
NOAA tide gauge at Fort McHenry and calculating the
Back Rivertide by applyingthe derived amplitude and
phase correction.
B.2.3 Description of One-Dimensional,
Cross-Sectionally Averaged Model
The numerical model which was used to simulate the
Back River hydrodynamics is an adaptation of
Hunter's one-dimensional model (Hunter 1975) as it
was applied to the Chesapeake and Delaware (C&D)
Canal. The model computes tidal elevation, flow,
salinity, and contaminant concentrations at interior
points given assigned boundary values and interior
sources and sinks. The model output was used as a
correction to field measurements.
The computational algorithm is based on a finite
difference representation of the momentum and
continuity equations. Non-advective transport iscon-
Figure B-1. Map showing the Back River segmentation scheme and water sampling locations.
Witchcoat
" Point
Porter
Point
Dye Smaplmg Transects
Boundary for Compouter Model Segment
® Water Sampling Location
B-2
-------
Table B-1. Dye Plume Mappings (Transects and Times)
Sampling Station
Date
1 1 Mar 84
13 Mar 84
15 Mar 84
17 Mar 84
20 Mar 84
2A
1416
1143
1322
1536
0946
1143
1301
1417
1610
1224
1430
3
1451
1154
1330
1544
0955
1154
1310
1432
1619
1239
1422
4
1504
1211
1341
1552
1005
1210
1324
1447
1637
1256
1414
5
1515
1222
1351
1602
1019
1220
1334
1459
1646
1312
1354
6
1541
1238
1404
1613
1031
1234
1353
1511
1701
1330
1339
7
1602
1251
1411
1631
1036
1300
1414
1535
1759
1346
1325
8
1250
1402
1622
1027
1250
1405
1525
1751
1144
1313
9
1240
1352
1613
1018
1236
1355
1515
1742
1127
1255
10
1225
1338
1558
1003
1223
1340
1500
1730
1107
1234
11
1214
1325
1544
0950
1213
1326
1447
1718
1045
1220
trolled by an exchange coefficient which is itself a
function of the hydraulic radius, Manning's "n," and
a single-valued diffusion factor which is used to
calibrate the model to observed data.
The model requires that the river be subdivided into
sections, the sizes of which are constrained by the
stability condition that the relation between the
section lengths (AX) andthe computational time step
(At) consistent with the following
conditions. Vertical measurements on 17 March
confirmed the validity of this assumption.
Freshwater inflow to the Back River is dominated by
the treatment plant flow. Surface run-off averages
less than 0.2 mVsec, whereas typical plant flows are
3 or 4 mVsec. For this reason, river flow was
neglected and hourly values of plant flow were input
into Section 1 of the model.
At<
AX
(Equation B-2) B_2.5 Calibration of Model
where g is the acceleration due to gravity and D is
river depth. The Back River was divided into seven
sections 1,600 m long which allows a time step of
300 seconds.
Geometric data for the model schematization were
taken from NOAA chart 12278. Required input
includes "typical" values of total surface width,
channel width, and depth for each section. The
"typical" values of width were derived by averaging
the widths from one-half a space step upstream to
one-half a space step downstream. Total surface
width includes side embayments; channel width does
not. These side embayments act as storage areas only
and do not directly participate in the transport of
momentum. The dye concentration data were aver-
aged over the cross section at each of the transects.
To do this, each transect was divided into 20
segments, and the chart recording of dye fluorescence
was also divided into 20 segments. The sum of the
products of the segment areas and the dye concen-
trations divided by the total cross-sectional area
yielded the cross-sectionally averaged dye concen-
tration as required by the model. This procedure
assumes that the dye is vertically mixed which is to be
expected in shallow water with March weather
Back River is only about 12 km in length which is
much shorter than a tidal wavelength for the domi-
nant Ma constituent. This makes it very difficult to
calibrate a model for hydrodynamic response, be-
cause tide gauges and/or current meters are not able
to resolve the slight differences caused by changes in
Manning's "n," which is the only parameter available
for hydrodynamic calibration. In lieu of a calibration
based on field data, Manning's "n" was set to 0.020,
which is the value that was used when this model
was applied to the C&D Canal and for a similar model
of the Potomac River where field data were used for
calibration.
The mixing and flushing characteristics of the model
are adjusted by two parameters—the diffusion factor
and the distance assigned to the "oceanic" source of
the contaminant. The diffusion factor is used in
calculating exchange coefficients as discussed above.
The distance to the "oceanic" value of the contami-
nant is a length scale used in a model algorithm for
predicting the influx of contaminant on the flood tide.
The term "oceanic" refers to a reservoir of constant
contaminant concentration.
Salinity was not included in the model because a
sensitivity test indicated that salinity contributions
B-3
-------
are not significant for salinity values at the mouth
between 0 and 1 5 ppt.
The best fit to the observed dye data was obtained
with the diffusion factor set at 1 50 and the distance to
the "oceanic" source set at 10 km (approximately one
tidal excursion).
B-4
-------
Appendix C
Biological Survey Sampling and Analytical Methods
C.1 Plankton Survey
Oblique bottom and near surface tows were'made at
eight stations in Back River and Middle River (Figure
2-1) using a double sled fitted with two 505-yum mesh,
0.5-m nets. The sled was towed for 5 minutes at each
depth for a total of 10 minutes. Tows were made only
near surface at shallow stations. AGeneral Oceanics
Model 2030 digital flowmeter was mounted in the
mouth of each net and a third one was mounted on
the sled outside the net to facilitate detection of net
clogging or meter malfunction. Tows were made
against the current. Each sample was placed in a
labeled 945-ml (1 -qt) jar and preserved in 10 percent
buffered formalin.
Water quality measurements consisting of tempera-
ture, dissolved oxygen, pH, and conductivity were
taken concurrently with plankton sampling at each
station.
Samples were examined in the laboratory under a
dissecting microscope and all macrozooplankton,
except the copepods, were enumerated, sorted into
major taxonomic groups, and preserved in 75 percent
ethanol for later identification. All organisms were
identified to the lowest practical taxon and counted.
Copepod densities were so high that subsampling
was required on all samples. Eurytemora affinis was
the only species of copepod observed in the sub-
samples. Depending on sample density, the sample
was either split with a Folsom plankton splitter, or
1.0- or 2.0-ml aliquots were taken with a Hensen-
Stempel pipette. Each subsample was put into a Ward
counting wheel and all copepods were counted. If
necessary, additional subsamples were examined
until at least 400 individuals were enumerated.
The number of copepods in the examined subsample,
the volume of subsamples examined, and the adjusted
volume of sample from which the subsamples were
taken were recorded so that organism number could
be converted to organism density during the initial
phases of data tabulation. Density was determined
from the equation
D = n(Vs/Va) / K(R, - R,) (Equation C-1)
where
D = number of organisms/100 L (density)
n = number of organisms counted in aliquot
Vs = volume of diluted sample
Va = volume of aliquot
Rf = final flowmeter reading
R, = initial flowmeter reading, and
K = flowmeter calibration factor (100 L/count).
This calculated density was used in all later data
analyses.
C.2 Benthic Macroinvertebrate Survey
A petite Ponar grab sampler (232 m2) was used to
collect three replicate samples at each station.
Samples were washed in the field through a No. 30
mesh screen (595 //m) to remove fine silt and clay
particles, placed in 945-ml labeled jars, and preserved
in 10 percent buffered formalin.
Water quality measurements consisting of tempera-
ture, DO, pH, and conductivity were taken concur-
rently with benthos sampling at each station. Quali-
tative determinations of the sediment type were also
made at each station.
Samples were sorted in the laboratory with the aid of
a dissecting microscope. Organisms were enumer-
ated, sorted into major taxonomic groups, and
preserved in 75 percent ethanol for later identifica-
tion. All organisms were identified to the lowest
practical taxon using appropriate keys and references.
Oligochaetes and chironomid larvae were mounted
on microslides prior to identification.
C.3 Fish Survey
Fish were collected at six stations in Back River and at
two reference stations in Middle River (Figures 3-1
and 3-2). At each station, a 4.9-m wide (16-ft) otter
trawl was towed at 1 m/sec for 10 minutes (600
meters). Specimens were identified and counted. Up
to 20 specimens of each species were also examined
closely for morphological anomalies, evidence of
diseases, and for parasites. This level of study
included examination of the gills, arches, and the gill
cavity surfaces. Additional specimens, if available,
were only examined grossly, i.e., the gill cavity was
not opened. Water quality parameters were also
reported.
C-1
-------
The number of specimens of each species was tallied
by station. The variety of abnormalities was listed,
and the incidence of conditions among the examined
specimens was determined for several species.
C-2
-------
Appendix D
Effluent Fractionation and Toxicity Testing Methods
D.1 Sampling
An effluent fractionation procedure was used to
detect toxic constituents in the effluents of the
Patapsco and Back River POTWs. Two composite
effluent samples, one a 3-day composite, and one a
7-day composite, were analyzed from each plant,
resulting in a total of four samples. The composites
were 19 L(5 gal) each in volume. The 3-day and 7-day
composites were initiated on the same day.
D.2 Ceriodaphnia Culture,
Maintenance, and Testing
Ceriodaphnia dubia was cultured in EA's labora-
tory in moderately hard reconstituted water (Table
D-1) spiked with 7 ml of 5 g/L yeast solution per liter
of water four days prior to usage. Cultures were kept
on a 16-hour light, 8-hour dark photoperiod at 25°C in
an environmental chamber and are fed a solution of
yeast and cerophyll daily, then thinned as necessary
to maintain healthy, productive, cultures. Adults from
these cultures were separated into lots of 300 at least
one day prior to test initiation and put in 1 -L culture
bowls and fed heavily. The morning of the test, gravid
adults were separated into lots of 100 and put into
4.5-in. culture dishes and fed. This ensured that
neonates used were of a specified age, preferably less
than 8 hours. During testing, organisms were fed 2
drops of yeast solution per cup.
Dilution water for test solutions was moderately hard
reconstituted water spiked with yeast four days prior
to testing. This water also served as control water.
Table D-1. Formulation for Moderately Hard Reconstituted
Water and Final Water Quality Ranges
Reagent Added (mg/liter)
NaHCO3
96
CaSO4-2H20
60
MgS04
60
KCI
4
Acute lethality tests lasting 48 hours were performed
in 1 -oz portion cups using the following test concen-
trations: 1.0, 3.0, 10.0, 30.0, and 100.0 percent plus a
dilution water control. Each concentration had 10
replicates with one organism per replicate. Effluent
and diluent were filtered through a 100-yum mesh to
remove large particles or any organisms that may be
present. Final volumes of 180 ml were mixed in
250 ml Class A graduated cylinders. Small volumes of
effluent were first measured in Class A pipettes, then
added to the graduate and brought to volume with
dilution water. The entire 180 ml of test solution was
poured into a dispenser calibrated to deliver 10
separate 15-ml portions. Neonates were then ran-
domly added, one per cup.
Water quality determination was performed on the
following schedule: pH, alkalinity, hardness, and
conductivity at sample receipt; pH, DO, and tempera-
ture at each renewal on one replicate control, low,
medium, and high test concentrations. Test vessels
were kept at 25 + 2°C on a 1 6-hour light, 8-hour dark
photoperiod cycle at a light intensity of 50 f.c.
Analytical methods were conducted according to
APHAetal. (1980).
D.3 Microtox®
The Microtox® test is a luminescence inhibition test
based on the proportionality between the light
produced by a luminescent marine bacterium (Photo-
bacterium phosphoreum) and its general respiratory
metabolism. Toxic effects of chemicals which include
reduction of metabolic rates are reflected in an
attenuation of the bioluminescence of the bacteria.
The bioluminescence response of the bacteria is
quantified by a photometer in the Microtox® unit. The
methods used for the Microtox® test followed those
found in the Beckman Microtox® instruction manual.
Final Quality
pH""
7.4-7 8
Hardness""
80-90
Alkalinity""
60-70
"Approximate pH after equilibrium.
"Expressed in mg/liter as CaCOa
D.4 Chemical Fractionation
To allow testing of the individual fractions of the
effluents, the chemical fractionation procedure of
Walsh and Garnas (1983) was followed (Figure D-1).
The effluent was filtered through a prewashed
Gelman Type A-E 1 -fjm pore size glass fiber filter to
D-1
-------
Figure D-1.
Fractionation and testing procedure.
remove solids, then eluted through a column of Rohm
and Haas Amberlite XAD-4 resin.
The inorganic fraction included all chemicals not
absorbed by the XAD-4 resin, which passed through
with the aqueous effluent. Before use, the resin was
prepared by repeated rinsing with deionized water, a
30-minute wash with 2 normal H2S04, and a final
de-ionized water rinse. Impurities were removed
from the resin by rinsing with technical-grade
acetone, followed by 1 2-hour sequential extractions
with acetone and methanol in a Soxhelet extractor.
XAD-4 column consisted of a 50-cc glass syringe,
loosely plugged with glass wool, and filled with 50 ml
(wet volume) of resin. At least 20 bed volumes of
distilled water were used to displace the methanol
from the column. A bored No. 6 teflon stopper coupled
to a 3-cm piece of 8-mm outside diameter tubing was
connected to the top of the column. Columns were
prepared in advance and stored in a refrigerator until
use.
During filtering, the 1-/wm glass fiber filter mounted
on a 142-mm filter holder, was fitted with a 2Q-/jm
nitex mesh prefilter to prevent clogging the glass fiber
filter.
The aqueous inorganic fraction from the XAD-4 resin
column was tested for toxicity following the proce-
dures outlined in Sections D.2 and D.3. If toxicity was
demonstrated, the inorganic fraction was further
fractionated into anion and cation fractions. This was
accomplished by a batch extraction procedure
whereby a 4-L sample of water was adjusted to pH >
10 and stirred for 24 hours with Dowex 1-X8 strong-
base anion-exchange resin at a level of 10 gm dry
resin/L water, to generate the cation fraction or
adjusted to pH < 4 and exposed to Dowex 50W-X8
strong-acid cation exchange resin to generate the
anion fraction. Following treatment, the resin was
removed from the sample by filtering through a glass
fiber filter, and the pH was adjusted to neutrality.
The whole organic fraction was considered to be the
fraction eluted from the XAD-4 resin column. This
was accomplished by aspirating the column to remove
excess water. The column was then eluted with
1 50 ml of nanograde acetone into a K-D concentrator
flask. The resultant sample was concentrated to 25
ml under vacuum at room temperature and an aliquot
was tested for toxicity using the methods described in
Sections D.2 and D.3. If toxicity to the whole organic
D-2
-------
fraction was found, further fractionation was per-
formed by separating the base/neutral and acid/
extractable subfractions following U.S. EPA Method
625 (U.S. EPA 1979) for priority pollutants. Prior to
toxicity testing with these subfractions the methylene
chloride was solvent exchanged with dimethyl sul-
foxide (DMSO).
D-3
-------
Appendix E
Toxicity Test Data
Table E-1. Routine Chemistry Data for the Ambient Tests, Baltimore Harbor, Maryland
Initial DO
(mg/L)
Ambient Station
Back River
B1
B2
B3
B4
B5
B6
Patapsco
P1
P2
P3
Middle River
Ml
M2
pH
69-7.5
6.9-7.5
68-7.5
6.9-7.4
7.0-7 7
7.0-8.0
68-7.5
69-7.4
6.8-7.4
69-7.1
6.8-7.2
Mean
8.3
8.5
8.6
8.5
87
8.6
8.4
8.5
8.5
8.7
86
Range
7.5-8.9
7.8-8.8
8.3-9.0
78-8.9
8.3-90
8.2-8.8
8.0-8 8
81-8.8
8.0-8.8
81-9.0
8.2-8.9
Final DO
(mg/L)
Mean
55
5.1
5 1
5.5
6.4
7.1
66
6.4
6.5
6.4
6.8
Range
2.3-8 4
1 9-84
2.9-7.7
4.3-8.0
4.9-9.1
58-10.4
6.0-7.4
5.6-7.3
59-71
5.5-75
5.6-7.6
Conductivity
(//mhos)
Mean
1,429
1,451
1,464
1,568
2,043
2,779
1,369
1,329
1,350
2,343
2,571
Range
1,250-1,650
1,300-1,700
1,300-1,600
1,350-2,300
1,650-2,800
2,200-3,500
1,150-1,500
1,100-1,550
1,250-1,500
2,250-2,600
2,000-3,000
Mean Salinity
(ppt ± SD)
0.5 ±0.25-0.75
0.57 ±0.5-0.75
0.64 ±0.5-0.75
0.68 ±0.5-1.0
1.0 ±0.75-1.5
1.5 ±1.0-2.0
8 ±6.5-88
79 ±6.3-8.6
8.0 ±7.0-9.0
1.0 ±1 0-1.1
1 3 ±1.0-1.5
Table E-2. Routine Chemistry Data for the Effluent Dilution and Salinity Tests
Initial DO
(mg/L)
Test
Back River
POTW
Concentration
100""
30
10
3
1
Control
pH
68-7.1
7.1-7.4
7.3-7.7
7.3-77
7.3-79
72-7.6
Mean
8.6
8.5
8.5
8.4
84
84
Range
86-8.7
8.2-8.9
82-8.8
8.2-8 7
80-8.7
8.1-8.6
Final DO
(mg/L)
Mean
5 1
5.3
56
6.1
6.1
6.2
Range
3.6-7.6
34-73
4.2-7.5
3.8-77
4.4-7.6
Conductivity
(,umhos)
Mean
925
666
581
508
494
477
Range
900-950
610-700
480-600
470-575
490-575
470-480
Mean Salinity
(ppt ± SD)
028 ±0.25-0.3
Patapsco
POTW
100""
30
10
3
1
Control
6.6
6.6-6.9
70-7.3
7.1-7.7
7.2-78
7.2-8.0
82
8.2
84
8.5
8.5
8.4
8 0-8.4
7.4-8.7
8.0-8.8
8.1-88
81-88
8.1-8.8
6.3
5.7
62
65
65
62
48-6.4
4.8-75
51-7.6
52-7.5
50-6.9
2,175
1,056
723
614
546
493
2,150-2,200
950-1,150
700-725
600-625
475-600
470-550
0.31 ±0.25-0.5
Salinity 16lbl 69
12 7.0-71
8 7.1-7.7
4 72-7.8
2 7.3-77
C 72-8.0
8.9
8.8
8.2
83
82
8.4
—
86-89
6.6-8 6
80-8.6
8 0-8.4
8.1-88
82
8.2
73
7.1
7.2
67
—
73-8.2
6 5-8.3
62-83
6.4-8 3
5.4-7.3
26,000
19,750
13,750
6,938
3,831
493
—
19,500-20,000
11,500-13,500
6,000-7,500
3,700-4,300
470-550
Concentrations are in percent
""Concentrations are in parts per thousand (ppt).
Note: Reconstituted water was used for dilution in all tests.
E-1
-------
Table E-3. Final Dissolved Oxygen Levels for Ceriodaphnia
dubia Effluent. Ambient, and Salinity Tests.
Baltimore Harbor, Maryland
Sample
Effluent
Patapsco POTW
Back River POTW
Percent
Effluent
(v/v)
100
30
10
1
3
Control
3.0
1.5
075
0.37
Control
100
30
10
3
1
Control
Mean DO
(mg/L)
7.2
7.3
75
78
7.6
76
74
77
74
7.5
68
6.8
76
6 1
76
7.5
DO
Range
—
—
—
73-78
73-8.0
7.5-7.9
7.3-8.0
7.2-8.1
5.3-7 2
7.4-7.6
73-76
7.1-8.0
70-79
Ambient
Back River
B1
82
B3
B4
B5
B6
Patapsco
P1
P2
P3
Control
Middle River
M1
M2
Salinity131
7.7
—
78
7.8
78
7.6
4
2
1
0.50
0.25
7.3
7.3
7.0
73
73
7.5
—
—
—
7.4-8.4
7.1-8.2
7.0-8.0
7.8
77
7.6
7.7
70-77
7.0-7.6
—
—
7.0-7.7
7 0-7.8
7.2-8.4
7 2-8.2
7 1-8.1
73-8.2
""Concentrations are in parts per thousand (ppt)
Note: Reconstituted water was used for dilution in all tests
f-2
-------
Appendix F
Biological Data
Table F-1. Results of a X2 Test Performed on the Number of Macrozooplankton Taxa, Back River, March 1984
Station
Number of taxa'al
Expected number (based on
average of M1 and M2)
X2 contribution"1'
B1
5
5.5
0
B2
8
55
072
B3
3
5.5
0 18
B4
3
5 5
0 18
B5
8
5 5
072
B6 M1
6 6
55
0
M2
5
'"'Number of unique taxa/life stages by combining two replicate samples for each station for two collection dates
lb'For individual station, the 1 degree of freedom X2 with P > X2 = 0.05 is 3 84
Note- For all stations combined, the calculated X2 = 3 18 (P > X2 = 0 78 with 6 d f)
X2 = (| E-01 |-05}a
Correction factor incorporated for
small (1 degree of freedom) dataset
Table F-2. Abundance [No./m3) of Macrozooplankton Collected from Back River and Middle River, 12 March 1984
Station
Station
Station
Taxa
E. affinis
M. edwardsi
Ostracoda
Chaoborus
B1L
63.693
—
—
—
B1R
45769
—
0016
0.016
Mean
54731
—
0008
0008
Station
Taxa
Daphnia
E. affinis
M edwardsi
Gammarus
Hemiptera IM
Nematoda
N americana
B4L
—
492 192
0052
—
—
—
—
B4R
0016
504.108
0037
—
__
—
__
Mean
0008
498 1 50
0044
—
__
—
—
Station
Taxa
Daphnia
E. affinis
M edwardsi
M1L
0.225
791.957
—
MIR
—
1,429.329
—
Mean
0.112
1,110.643
—
B2L
1 05 649
0021
—
—
B2R
103.423
—
—
0008
Mean
104.536
0010
—
0008
Station
B5L
0014
290 704
0240
0014
0.014
—
0014
B5R
—
274.229
0120
0031
—
0.016
—
Mean
0007
282.466
0.180
0.022
0007
0008
0007
B3L B3R
201492 215.362
0.031
—
0016
Station
B6L B6R
0.029
1,118296 952334
0233 0299
—
__
—
—
Mean
208.427
0.015
—
0.008
Mean
0014
1,035.315
0.266
—
—
—
—
Station
M2L
0021
385.541
0099
M2R
—
391 545
0054
Mean
0.010
388 543
0076
F-1
-------
Table F-3. Abundance (No./m3) of Macrozooplankton Collected from Back
Station
Taxa
E. affinis
M. edwardsi
Ceriodaphnia
Gammarus
Ostracoda
Chironomidae
P.
Diptera P.
Chaoborus
B1L
15.243
0.022
0.063
—
—
—
—
—
B1R
22459
0.016
—
—
—
—
—
—
Mean
18.851
0019
0.032
—
—
—
—
—
Station
Taxa
Daphnia
E. affinis
M. edwardsi
Gammarus
Eubosmina
N. americana
L, plumulosus
B4L
—
368 905
—
—
—
—
—
B4R
—
350.774
0016
—
—
—
—
Mean
—
359 840
0.008
—
—
—
—
Station
Taxa
Daphnia
E. affinis
M. edwardsi
Collembola
Eubosmina
Diptera P.
A. proximoculi
Chaoborus
M1L
0.578
1 ,460.698
—
0.014
0.014
0014
—
—
M1R
0.265
1,180.544
—
0.017
—
—
0017
—
Mean
0.422
1,320.621
—
0.016
0.007
0.007
0008
—
Station
B2L
117.916
0.056
—
0.023
0.014
0.014
0014
—
B2R
142.137
—
0.039
—
—
—
—
0.016
Station
B5L
—
936 747
0.061
0.014
—
—
—
B5R
0.076
1,040.629
0054
—
0.016
—
—
River and Middle River, 16 March 1984
Station
Mean B3L B3R
130.026 50.565 65.941
0.028 0.048 0.063
0.020
0.012
0.007
0.007
0.007
0.008
Station
Mean B6L B6R
0.038 0.014 0.023
988.688 272.122 235.941
0.058 0.044 0.073
0.007 — 0.030
0.008
0014
0.023
Mean
58.253
0.056
—
—
—
—
—
—
Mean
0.018
254.032
0.058
0.015
—
0.007
0.012
Station
M2L
0.847
290.488
0.039
—
0.014
—
—
0.022
M2R
0.831
341.032
0.050
—
0.017
—
—
—
Mean
0839
315.760
0.044
—
0.016
—
—
0.011
Table F-4. Analysis of Variance and Tukey's Studentized
Range Test Results for Eurytemora affinis. Back
River, March 1984
Dependent Variable- In density (No./m3)
Source Df Squares Square F-Value PR > F
Treatment
Date
Station
Date x station
Error
Corrected Total
15
1
7
7
16
31
43.41
0.82
3696
562
0.42
43.83
289
0.82
528
0.80
2.89
002
111.55
31.84
203.52
30.97
00001
0.0001
0.0001
0.0001
Tukey's Studentized Range Test on Station Abundances
Station M1 B5 B6 B4 M2 B2 S3 B1
Mean 1n count (7.1) (6.3) (6.2) (6.1) (5.9) (4.8) (47) (35)
F-2
-------
Table F-5. Water Quality Data from Back River and Middle River, 12 and 16 March 1984
Station
12 March 1984
B1
B2
B3
B4
B5
B6
M1
M2
16 March 1984
B1
B2
B3
B4
B5
B6
M1
M2
Station
12 March 1984
B1
B2
B3
B4
B5
B6
M1
M2
1 6 March 1 984
B1
B2
B3
B4
B5
B6
M1
Time
1630
1607
1543
1443
1351
1313
1140
1225
1533
1512
1438
1353
1258
1225
1045
1125
Time
1630
1607
1543
1443
1351
1313
1140
1225
1533
1512
1438
1353
1258
1225
1045
Depth
(m)
0.3
0.3
1.3
1.0
1.0
2.5
3.0
3.0
0.3
1.0
1.0
1.5
2.5
2.0
3.0
2.5
Tide
F
F
F
F
F
F
F""
F
F
F
F
F
LS
LS
E
Surface
0.8
0.9
0.7
0.9
1.2
2.1
1.4
1.7
0.6
0.7
0.7
0.7
1 2
1.6
1.3
1.5
Surface
5.3
5.1
4.3
35
2.8
2.1
2.8
1.9
9.9
9.1
9.4
9.7
6.1
6.0
4.4
Salinity
Middle
—
—
—
—
—
1.4
1.8
__
—
—
07
1.2
1.6
1.3
1 5
Temperature
Middle
—
—
—
—
—
2.6
1.9
—
—
72
5.9
50
3.8
Bottom
—
0.7
0.9
1 2
2.1
1.4
2.1
__
07
0.7
0.7
2.5
25
1.4
1.6
Bottom
—
44
3.5
2.8
2.0
2.5
1.8
8.2
8.7
7.1
5 1
4.0
34
Surface
7.5
7.4
7.5
78
8.5
8.4
6.9
7.3
7.0
7.0
7.1
7.2
8.4
8.1
7.0
7.5
Surface
14.0
13.0
12.0
152
16.7
16.2
132
14.8
8.9
6.3
6.5
7.1
174
17.4
14.2
pH
Middle
—
—
—
—
—
7.2
7.5
—
—
7.1
8.6
8.6
7.2
7.9
DO
Middle
—
—
—
—
—
12.6
133
—
—
9.9
16.6
15.6
13.7
Bottom
—
7.3
7.8
8.5
8.4
7.1
7.5
6.9
6.9
7.1
8.3
8.3
7.1
7.8
Bottom
__
—
11.0
14.1
15.6
15.0
128
13.6
__
62
6.0
9.8
14.6
13.6
13.4
"F = Flood, E= Ebb, LS = Low slack.
F-3
-------
Table F-6. Results of a X2 Test Performed on the Number of Benthic Macroinvertebrate Taxa, Back
Station
Number of taxalal
Expected number (based on
average of M1 and M2)
X2 contribution
B1
4
11 5
4 26
B2
5
11 5
3 13
B3
3
115 1
5 56
B4 B5
2 8
15 115
7 04 0 78
B6
12
11 5
0
River, March 1984
M1
10
M2
13
""Number of unique taxa/life stages by combining three replicate samples for each station for two collection dates
lb'For individual station, the 1 degree of freedom X2 with P > X2 = 0 05 is 3 84
Note- For all stations combined, the calculated X2 = 23 77 |P > X2 = 0 005 with 6 d f )
X2 - 1 1 E-O I -0 5)2
>l ~ l~ ' Cnrrp.rtinn far.tnr inrnrpnratfirHnr O - Ohsprvpr)
E small (1 degree of freedom) dataset. E = Expected
Table F-7. Water Quality Data from Back River and Middle River, 1 9 March 1 984
n^nth Salinitv
Station
B1
B2
B3
B4
B5
B6
M1
M2
Station
B1
B2
B3
B4
B5
B6
M1
M2
Table F-8.
Number of taxa
Time
1435
1405
1330
1300
1130
1050
0905
1000
Time
1435
1405
1330
1300
1130
1050
0905
1000
Results of a X
(a)
(m)
1 5
1 0
20
20
30
30
30
40
Tide
1 5
1 0
20
20
30
3.0
30
40
Surface
06
07
07
09
1 4
2 6
1 3
07
Surface
11 2
11 0
10 1
94
82
64
64
55
Middle
—
08
09
2 1
34
1 3
23
Temperature
Middle
—
87
90
63
50
63
49
Bottom Surface
07
08
08
1 2
2 1
3 5
1 3
27
7.
7
7.
8
8
8
7
7
6
6
8
2
0
2
1
8
Bottom Surface
97
85
8 7
74
6.2
49
65
48
8
9
pH
Middle
—
—
74
7 9
84
81
72
79
DO
Middle
Bottom
7
7
7
7.
8
8
7
7
5
5
4
9
2
1
1
9
Bottom
9.0
94
1 1
3
149
13
13
11
13
2 Test Performed on the Number of Fish Taxa, Back River, March
B1
3
B2
2
B3
1
Station
B4 B5
4 3
9
9
7
2
1984
B6
4
106
13.8
139
132
11 4
128
M1
7
9
11
12
13
13
11
12
6
.0
4
8
2
.0
8
M2
5
Expected number (based on
average of M1
X2 contribution
and M2)
""Number of unique taxa/life
""For individual
station, the 1
6
1 04
6
204
stages by combining samples
6
3.38
6 6
0 38 1 04
from two collection dates for each
6
038
—
—
-
-
-
-
station.
degree of freedom X2 with P > X2 = 0.05 is 3.84.
Note: For all stations combined, the calculated X2 = 8 01 (P
>X2= 0.240 with 6df ).
X =(|E-O|-05) Correction factor incoraorated for O =
E
small (1 degree
of freedom) dataset. E =
Observed
Expected
F-4
-------
Table F-9. Trendsin Abnormalities Observed Among Brown Bullheads Collected in Back River and Middle River, 7 March 1984
Station
Observation
Muscular atrophy
Healed/healing scars
Nodule/tumor
Spinal curvature (lordosis)
Unusual coloration
Small whitish spots
Small dark spots
Fin erosion/rot
Regenerated fin/rays
Missing fin
Gill filament erosion
Gill arch cyst
Blind eye
Number examined closely
Number examined grossly
Total
B1 B2
16.7%
(1/6)
8.3%
(1/12)
167% 8.3%
(1/6) (1/12)
8.3%
(1/12)
8.3%
(1/12)
6 12
0 0
6 12
B3 B4
08%
(1/126)
0.8%
(1/126)
0.8%
(1/126)
0.8%
(1/126)
08%
(1/126)
0.8%
(1/126)
1 .6%
(2/126)
1 .6%
(2/126)
5.9%
(1/17)
0.8%
(1/126)
17 20
0 106
17 126
B5
29%
(2/69)
2.9%
(2/69)
1 .4%
(1/69)
14
55
69
B6
N
O
A
B
N
0
R
M
A
L
I
T
I
E
S
1
0
1
B1-B3
2.9%
(1/35)
2.9%
(1/35)
5.7%
(2/35)
29%
(1/35)
2.9%
(1/35)
2.9%
(1/35)
35
0
35
B4-B6
1.5%
(3/196)
0.5%
(1/196)
05%
(1/196)
05%
(1/196)
0.5%
(1/196)
0.5%
(1/196)
2.0%
(4/196)
1 .0%
(2/196)
1 0%
(2/196)
35
161
196
Table F-10.
Trends in Abnormalities Observed Among Brown Bullheads Collected in Back River and Middle River, 14 March
1984
Station
Observation
Healed/healing scars
Nodule/tumor
Fin erosion/rot
Regenerated fins/rays
White cysts on fins
Black cysts on fins
Blind eye
Number examined closely
Number examined grossly
Total
B1
5 1%
(2/39)
1 0.3%
(4/39)
2.6%
(1/39)
2.6%
(1/39)
20
19
39
B2
N
O
A
B
N
0
R
M
A
L
I
T
I
E
S
2
0
2
B3
40%
(1/25)
1 2.0%
(3/25)
40%
(1/25)
20
5
25
B4 B5
26%
(1/39)
2.1% 51%
(1/48) (2/39)
2.6%
(1/39)
18 20
30 19
48 39
B6
N
0
C
A
T
C
H
—
0
B1-B3
1 .5%
(1/66)
7.6%
(5/66)
7.6%
(5/66)
1.5%
(1/66)
1 5%
(1/66)
42
24
66
B4-B6
1 1%
(1/87)
34%
(3/87)
1.1%
(1/87)
38
49
87
F-5
-------
Table F-11. Trends in Abnormalities Observed Among White Perch Collected in Back River and Middle River, 7 March 1984
Station
River
Observation
Body lesions
Body fungus — smooth, opaque
slime
Fin erosion/rot
Regenerated fin/rays
Gill filament erosion
Gill raker erosion
Blind eye
Ergasilus
Leech
Number examined closely
Number examined grossly
Total
B5
20 0%
(2/10)
30 0%
(3/10)
10
0
10
B6
27%
(2/74)
1 .4%
(1/74)
2.7%
(2/74)
5.0%
(1/20)
1 .4%
(1/74)
55 0%
(11/20)
1 4%
(1/74)
20
54
74
M1
3.7%
(1/27)
3.7%
(1/27)
65 0%
(13/20)
11 1%
(3/27)
20
7
27
M2
1 4 3%
(1/7)
28.6%
(2/7)
7
0
7
Back
24%
(2/84)
1 2%
(1/84)
2.4%
(2/84)
3.3%
(1/30)
6.7%
(2/30)
1.2%
(1/84)
46 7%
(14/30)
1.2%
(1/84)
30
54
84
Middle
2.9%
(1/34)
2.9%
(1/34)
29%
(1/34)
55 6%
(5/27)
8.8%
(3/34)
27
7
34
Table F-12. Trends in Abnormalities Observed Among Pumpkinseed and White Perch Collected in Back River and Middle River,
14 March 1984
Pumpkinseed
White Perch
Observation
Muscular atrophy
Nodule/tumor
Deformed jaw
Pughead
Fin erosion/rot
Regenerated fins/rays
Gill filament erosion
Pale gill filaments
Ergasilus
Leech
Gill raker erosion
Blind eye
Number examined closely
Number examined grossly
Total
B4-B6
N
0
A
B
N
0
R
M
A
L
I
T
I
E
S
4
0
4
M1
2%
(1/50)
2%
(1/50)
2%
(1/50)
2%
(1/50)
6%
(3/50)
14%
(7/50)
5%
(1/20)
20%
(4/20)
30%
(15/50)
20
30
50
M2
25%
(1/4)
25%
(1/4)
50%
(2/4)
25%
(1/4)
4
0
4
Back River
N
O
A
B
N
0
R
M
A
L
I
T
I
E
S
4
0
4
Middle River B6
1 9%
(1/54)
1 9%
(1/54)
1 9%
(1/54)
1.9%
(1/54)
56%
(3/54)
14.8%
(8/54)
4.2%
(1/24)
42%
(1/24)
25 0% 1 5.0%
(6/24) (3/20)
29 6% 5 7%
(16/54) (2/35)
24 20
30 15
54 35
M1
400
(4/10)
10.0%
(1/110)
1 0 0%
(1/10)
20 0%
(2/10)
10
0
10
f-6
-------
Table F-13. List of Fish Species and Families Collected,
Back River and Middle River, March 1984
Family
Cyprmidae
(minnows)
Centrarchidae
(sunfish)
Percichthyidae
(temperate
basses)
Percidae
(perches)
Ictaluridae
(catfish)
Clupeidae
(herring)
Gasterasteidae
(sticklebacks)
Scientific Name
Notropis spilopterus
Lepomis gibbosus
Morone americana
Perca flavescens
Ictalurus nebulosus
Ictalurur punctatus
Alosa aestivalis
Drosoma cepedianum
Gasterosteus
wheatlandi
Common Name
Spotfin shiner
Pumpkinseed
sunfish
White perch
Yellow perch
Brown bullhead
Channel catfish
Blueback herring
Gizzard shad
Blackspotted
stickleback
F-7
-------
Appendix G
Support Chemical Fractionation Data
The results of the acute Ceriodaphnia dub/a 48-hour
LC50 tests for the Back River and Patapsco POTW
effluents were discussed in Chapter 11, as part of the
effluent fractionation procedure tests. The mortality
data for these tests, m which 10 Ceriodaphnia were
exposed to various concentrations of whole effluent
and fractions derived from the effluent fractionation
procedure described in Appendix D, are presented in
Table G-1. As was discussed in Chapter 11, LCBOs
could not be calculated for certain of the tests,
because of the absence of partial kills, or because of
the absence of a valid dose-response relationship in
the data.
The results of the chemical tests on the base/neutral
subfraction of the organic fraction of the 3-day and
7-day composites of the Patapsco POTW effluents,
which were the subfractions which displayed much
of the toxicity observed in the samples tested, were
discussed m Chapter 1 2. The documentation for the
GC/MS analyses for the base/neutral priority pollu-
tants is presented in this Appendix (Tables G-2
through G-8 and Figures G-1 through G-8). Recon-
structed ion chromatograms and quantitation reports
are presented for the standard (Figure G-1, Table
G-2), the surrogate spike standard (Figure G-2, Table
G-3), and blank (Figure G-3, Table G-4). A quantitation
report is provided for the spike of the sample blank
(Table G-5). Reconstructed ion chromatograms and
quantitation reports are also provided for the 3-day
composite (Figure G-4 and Table G-6), and the 7-day
composite (Figure G-6 and Table G-7), while Figure
G-5 presents the results of a library search to obtain a
possible match for a compound noted in the 3-day
composite. Documentation of the DFTPPtuning ofthe
GC/MS is presented in Figures G-7 and G-8 and
Table G-8.
G-7
-------
Table G-1. Ceriodaphnia dubia Mortality in 48-Hour LC50 Tests on Back River and Patapsco POTW
3-Day and 7-Day Composite Samples
Percent Effluent (v/v)
Back River POTW 3-Day Composite
100
30
10
3
1
0 (control)
7-Day Composite
100
30
10
3
1
0 (control)
Percent Effluent (v/v)
Patapsco POTW 3-Day Composite
100
30
10
3
1
0 (control)
7-Day Composite
100
30
10
3
1
0 (control)
Whole
Effluent
10
6
8
6
5
3
10
2
5
3
1
2
Whole
Effluent
10
10
10
4
5
3
10
10
10
2
2
1
Inorganic
Fraction
Inorganic
Fraction
10
2
1
1
1
0
3
0
1
0
1
0
3
0
0
2
2
1
0
0
1
0
1
0
Cation
Fraction
10
0
0
0
0
0
—
—
—
—
—
—
Organic
Fraction
10
0
1
2
3
1
10
1
1
3
2
0
Anion
Fraction
2
2
0
0
0
0
—
—
—
—
—
—
Base/Neutral Acid/Phenol
Fraction
3
6
6
7
4
3
5
3
3
4
2
0
Organic
Fraction
10
10
2
2
0
1
10
10
0
1
2
2
Base/
Neutral
Fraction
10
10
4
4
5
0
8
6
8
3
1
2
Fracnon
5
5
5
8
4
2
5
1
0
3
1
0
Acid/
Phenol
Fraction
2
4
4
2
2
2
6
1
1
1
0
1
Note: Reconstituted water was used as dilution water
G-2
-------
Table G-2. Base/Neutral Standard Quantitation Report for 3-Day and 7-Day Patapsco POTW Base/Neutral Fraction Effluent
Analysis
Name
m/z
Scan
Time
Ref.
RRT
Meth. Area (Hght)
Amount
% Tot.
D-8 Naphthalene (I.S #1)
D1 0-Phenanthrene (I.S. #2)
D12-Chrysene(I.S. #3)
N-Nitrosodimethylamme
Bis(2-Chloroethyl)ether
1 ,3-Dichlorobenzene
1 ,4-Dichlorobenzene
1 ,2-Dichlorobenzene
Bis(2-Chloroisopropyl)ether
N-Nitroso-di-n-propylamine
Hexachloroethane
Nitrobenzene
Isophorone
Bis(2-Chloroethoxy)methane
1 ,2,4-Tnchlorobenzene
Naphthalene
Hexachlorobutadiene
Hexachlorocyclopentadiene
2-Chloronaphthalene
Dimethyl phthalate
Acenaphthylene
2,6-Dmitrotoluene
Acenaphthene
2,4-Dinitrotoluene
Diethyl phthalate
Fluorene
4-Chlorophenylphenyl ether
N-Nitrosodiphenylamme
1 ,2-Diphenylhydrazme
4-Bromophenylphenyl ether
Hexachlorobenzene
Phenanthrene
Anthracene
Di-n-butylphthalate
Fluoranthene
Benzidine
Pyrene
Butylbenzyl phthalate
3,3'-Dichlorobenzidme
Benzo(a)anthracene
Chrysene
Bis(2-Ethylhexyl)phthalate
Di-n-octyl phthalate
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(a)pyrene
lndeno(1,2,3-cd)pyrene
Dibenzo(a,h)anthracene
Benzo(g,h,i)perylene
136
188
240
74
93
146
146
146
45
70
117
123
82
93
180
128
225
237
162
163
152
165
154
89
149
166
204
169
77
248
284
178
178
149
202
184
202
149
252
228
228
149
149
252
252
252
276
278
276
1482
2615
3567
496
1040
1081
1098
1152
1198
1241
1240
1279
1255
1441
1481
1500
1563
1809
1899
2045
2045
2067
2112
2201
2295
2290
2299
2349
2353
2468
2509
2623
2640
2874
3049
3155
3126
3405
3578
3562
3578
3629
3855
3961
3973
4097
4738
4765
4918
10:50
18.59
2553
3.36
7 33
7:51
758
822
8-42
9:00
900
9.17
950
10:27
10.45
1053
11:21
1308
13:47
14 50
14-50
15:00
15.20
15.58
16 39
16.37
1641
17.03
1705
17 55
18 13
1902
19:10
2051
2208
2254
22:41
2443
2558
25 51
25:58
26.20
27:59
2845
28:50
29.44
3423
3435
3542
1
2
3
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
3
3
1.000
1.000
1.000
0332
0.697
0.725
0.736
0.772
0.803
0.832
0.831
0.857
0.908
0.966
0.993
1.005
1.048
1.212
1.273
1.371
1.371
1 385
0.908
0.842
0.878
0876
0.879
0.898
0.900
0.944
0959
1.003
1.010
1.099
1.166
1.207
1.195
0.955
1 003
0.999
1.003
1.017
1.081
1.110
1 1 14
1.149
1.328
1 336
1.379
A BB
A BB
A BB
A BB
A BB
A BV
a BV
A BB
A BB
A BB
A BB
A BB
A BB
A BB
A BV
A BB
A BB
A BB
A BB
A BB
A BB
A BB
A BB
A BB
A BB
A VB
A BB
A BB
A BB
A BB
A BB
A BV
A VB
A BB
A BB
A BB
A BB
A BV
A BB
A BV
A VB
A BV
A BB
A BV
A VB
A BB
A BB
A BB
A BB
1 640290.
422573.
161155
2326150.
2472650.
2740570.
2659490.
2406020.
3176600.
1242710.
1 1 33950.
668276
3226170.
1611600.
1456950.
5133090.
982232
286588.
1 994830.
2396960.
3688860.
247250
1677720.
121629.
2701970.
1734180
939726
668102.
2106620.
428470.
547807
1767770
2071970.
1676360.
1578040
588
1133110
267404.
87263.
663416
834147.
395568.
377100.
303478
320916.
201 020.
89024.
101606
83677.
20.000 ppm
20.000 ppm
20.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50 000 ppm
50.000 ppm
50.000 ppm
50 000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50 000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
50 000 ppm
50 000 ppm
50.000 ppm
50.000 ppm
50 000 ppm
50.000 ppm
50.000 ppm
50. 000 ppm
0.85
0.85
0.85
2.12
2.12
2 12
2.12
2.12
2.12
2.12
2.12
212
2.12
2.12
2.12
2.12
2.12
2.12
2.12
2 12
2.12
2.12
2.12
212
2.12
2.12
2 12
2.12
2 12
2 12
2.12
2.12
212
2.12
2.12
2.12
2.12
2 12
2.12
2 12
2.12
2 12
2 12
2.12
2.12
2 12
2 12
2.12
2.112
Table G-3. Surrogate Spike Standard Quantitation Report for 3-Day and 7-Day Patapsco POTW Base/Neutral Fraction Effluent
Analysis
Name
D-8 Naphthalene (I.S. #1)
D10-Phenanthrene (I S. #2)
2-Fluorophenol (A/P Surr )
D-5 Phenol (A/P Surr.)
05-Nitrobenzene (B/N Surr )
2-Fluorobiphenyl (B/N Surr.)
m/z
136
188
112
99
128
172
Scan
1490
2614
779
1034
1272
1874
Time
1049
1858
539
730
9-14
13 36
Ref
1
2
1
1
1
1
RRT
1.000
1.000
0.523
0694
0.854
1 258
Meth.
A BB
A BB
A BB
A BB
A BB
A BB
Area (Hght)
1284310
350927.
3086270
1 552090.
767043
2634180.
Amount
20.000 ppm
20.000 ppm
50.000 ppm
50.000 ppm
50.000 ppm
bo.000 ppm
% Tot.
8.33
833
2083
20.83
2083
2083
-------
Table G-4. Blank Quantitation Report for 3-Day and 7-Day Patapsco POTW Base/Neutral Fraction Effluent Analysis
Name m/z Scan Time Ref RRT Meth. Area (Hght) Amount
D-8 Naphthalene (1 S. #1)
D10-Phenanthrene(I.S #2)
D12-Chrysene (I.S #3—
N-Nitrosodimethylamine
Bis(2-Chloroethyl)ether
1,3-Dichlorobenzene
1 ,4-Dichlorobenzene
1 ,2-Dichlorobenzene
Bis(2-Chloroisopropyl)ether
N-Nitroso-di-n-propylamme
Hexachloroethane
Nitrobenzene
Isophorone
Bis(2-Chloroethoxy)methane
1 ,2,4-Tnchlorobenzene
Naphthalene
Hexachlorobutadiene
Hexachlorocyclopentadiene
2-Chloronaphthalene
Dimethyl phthalate
Acenaphthylene
2,6-Dmitrotoluene
Acenaphthene
2,4-Dmitrotoluene
Diethyl phthalate
Fluorene
4-Chlorophenylphenyl ether
N-Nitrosodiphenylamme
1 ,2-Diphenylhydrazine
4-Bromophenylphenyl ether
Hexachlorobenzene
Phenanthrene
Anthracene
Di-n-butylphthalate
Fluoranthene
Benzidme
Pyrene
Butylbenzyl phthalate
3,3'-Dichlorobenzidme
Benzo(a)anthracene
Chrysene
Bis(2-Ethylhexyl)phthalate
Di-n-octyl phthalate
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(a)pyrene
lndeno(1 ,2,3-cd)pyrene
Dibenzo(a,h)anthracene
Benzo(g,h,i)perylene
106 1490 1049 1 1000 ABB 595682 20 000 ppm
188 2612 1857 2 1.000 ABB 176072 20.000 ppm
240 3564 2552 3 1000 ABB 35150. 20.000 ppm
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
128 1490 1049 1 1000 ABB 2068 0 055 ppm
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
149 2870 20.50 2 1099 A BV 2492 01 78 ppm
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
% Tot
33.33
33.33
33.33
0.09
0.30
Table G-5. Spike Blank Quantitation Report for 3-Day and 7-Day Patapsco POTW Base/Neutral Fraction Effluent Analysis
Name
D-8 Naphthalene (I S. #1)
D10-Phenanthrene (I S. #2)
2-Fluorophenol (A/P Surr.)
D-5 Phenol (A/P Surr)
D5-Nitrobenzene (B/N Surr )
2-Fluorobiphenyl (B/N Surr )
m/z
136
188
112
99
128
172
Scan
1490
2612
783
1037
1273
1872
Time
10:49
1857
541
7.32
9 14
13:35
Ref
1
2
1
1
1
1
RRT
1 000
1 000
0.526
0696
0854
1 256
Meth
A BB
A BB
A BB
A BB
A BB
A BB
Area (Hght)
595682.
176072
46531.
192315.
704377.
2448540
Amount
20.000 ppm
20.000 ppm
1.625 ppm
13.357 ppm
98.995 ppm
100.205 ppm
% Tot.
7.87
7.87
0.64
5.26
3895
39.42
G-4
-------
Table G-6. Quantitation Report for 3-Day Patapsco POTW Base/Neutral Fraction Effluent Analysis
Name m/z Scan Time Ref RRT Meth Area (Hght) Amount % Tot.
D-8 Naphthalene (I.S #1)
D10-Phenanthrene (I S. #2)
D12-Chrysene (1 S. #3)
N-Nitrosodimethylamine
Bis(2-Chloroethyl)ether
1 ,3-Dichlorobenzene
1 ,4-Dichlorobenzene
1 ,2-Dichlorobenzene
Bis(2-Chloroisopropyl)ether
N-Nitroso-di-n-propylamme
Hexachloroethane
Nitrobenzene
Isophorone
Bis(2-Chloroethoxy)methane
1 ,2,4-Tnchlorobenzene
Naphthalene
Hexachlorobutadiene
Hexachlorocyclopentadiene
2-Chloronaphthalene
Dimethyl phthalate
Acenaphthylene
2,6-Dmitrotoluene
Acenaphthene
2,4-Dmitrotoluene
Diethyl phthalate
Fluorene
4-Chlorophenylphenyl ether
N-Nitrosodiphenylamme
1 ,2-Diphenylhydrazine
4-Bromophenylphenyl ether
Hexachlorobenzene
Phenanthrene
Anthracene
Di-n-butylphthalate
Fluoranthene
Benzidme
Pyrene
Butylbenzyl phthalate
3,3'-Dichlorobenzidme
Benzo(a)anthracene
Chrysene
Bis(20Ethylhexyl)phthalate
Di-n-octyl phthalate
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(a)pyrene
lndeno(1,2,3-cd)pyrene
Dibenzo(a,h)anthracene
Benzo(g,h,i)perylene
136 1488 1048 1 1000 ABB 271288 20000ppm 3313
188 2611 1857 2 1.000 A BV 123922 20 000 ppm 33.13
240 3566 2553 3 1000 ABB 25990 20 000 ppm 33.13
Not Found
93 1037 732 1 0697 ABB 2916 0 357 ppm 059
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
77 2344 1701 2 0898 ABB 4116 0.333 ppm 0.55
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
G-5
-------
Table G-7. Quantitation Report for 7-Day Patapsco POTW Base/Neutral Fraction Effluent Analysis
Name m/z Scan Time Ref. RRT Meth Area (Hght) Amount % Tot.
D-8 Naphthalene (1 S #1)
D10-Phenanthrene (I.S #2)
D12-Chrysene(I.S #3)
N-Nitrosodimethylamme
Bis(2-Chloroethyl)ether
1 ,3-Dichlorobenzene
1,4-Dichlorobenzene
1 ,2-Dichlorobenzene
Bis(2-Chloroisopropy!) ether
N-Nitroso-di-n-propylamme
Hexachloroethane
Nitrobenzene
Isophorone
Bis(2-Chloroethoxy)methane
1",2,4-Tnchlorobenzene
Naphthalene
Hexachlorobutadiene
Hexachlorocyclopentadiene
2-Chloronaphthalene
Dimethyl phthalate
Acenaphthylene
2,6-Dmitrotoluene
Acenaphthene
2,4-Dmitrotoluene
Diethyl phthalate
Fluorene
4-Chlorophenylphenyl ether
N-Nitrosodiphenylamme
1 ,2-Diphenylhydrazme
4-Bromophenylphenyl ether
Hexachlorobenzene
Phenanthrene
Anthracene
Di-n-butylphthalate
Fluoranthene
Benzidme
Pyrene
Butylbenzyl phthalate
3,3'-Dichlorobenzidine
Benzo(a)anthracene
Chrysene
Bis(2-Ethylhexyl)phthalate
Di-n-octyl phthalate
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(a)pyrene
lndeno(1 ,2,3-cdjpyrene
Dibenzo(a,h)anthracene
Benzo(g,h,i)perylene
136 1488 10.48 1 1 000 A BB
188 2609 18.56 2 1000 ABB
240 3563 25:52 3 1.000 A BV
Not Found
93 1038 732 1 0.698 A BB
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
7 2344 17:01 2 0898 A BB
Not Found
Not Found
Not Found
Not Found
149 2870 20-50 2 1.100 A BB
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
149 3622 26.17 3 1 017 A BB
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
Not Found
244822 20.000 ppm 30.16
130420 20.000 ppm 30.16
17752 20.000 ppm 30.16
4416 0598 ppm 0.90
1456 0.1 12 ppm 0.17
520. 0.050 ppm 0.08
5116. 5.871 ppm 8.85
G-6
-------
Table G-8. Mass List for DFTPP Analysis on 3-Day and 7-Day Patapsco POTW Base/Neutral Fraction Effluent
50
445
Mass
50.05 F
51 09 F
52.21 F
54.85 F
56.13 F
57.06 F
63.07 F
65.07 F
68.98 F
7391 F
75.09 F
77.02 F
79.11 F
80.06 F
81.06 F
8221 F
83.23 F
91.10 F
92.09 F
93.07 F
98.05 F
99.09 F
100.85 F
103.13 F
104.16 F
1 05 1 5 F
10702 F
108.08 F
110.03 F
111.09 F
116.95 F
121.85 F
123.10 F
1 24 1 2 F
127.05 F
128 14 F
129.08 F
13002 F
135.09 F
13720 F
141.16 F
14711 F
148.03 F
149 12 F
151.53
15508 F
156.14 F
157 64 F
159.17 F
161 20 F
0.00
%RA
15.20
35.93
2.30
1.68
1.80
3.94
1 62
1.02
41 98
5.43
6.98
46.36
2.53
213
2.97
0.97
1 06
0.98
1 18
4.05
292
2.73
2.37
0.98
0.98
0.97
11.83
1.99
25.58
3.12
969
1.22
1.37
1.00
41 91
3.41
1503
1.62
1 48
1.26
2.24
1 33
2.51
1.09
1 20
1 49
1.69
1.38
1.77
1.44
000
% RIC
1.87
443
028
0.21
022
0.47
0.20
013
5.18
067
0.86
5.72
0.31
0.26
0.37
0.12
0.13
0.12
0.15
050
0.36
034
0.29
0 12
0.12
0.12
1.46
0.25
316
0.38
1.20
015
0.17
0.12
5.17
0.42
1.85
0.20
0.18
0.16
0.28
0.16
031
0.13
0.15
0.18
021
0.17
0.22
0.18
2. Minima
0. Maxima Mm. Inten. 203
Inten. Mass % RA
3336
7888
504.
368
396.
844.
356
224.
9216.
1192.
1532.
10176.
556.
468.
652.
212.
232
216.
260.
888
640.
600.
520.
216
216.
212.
2596.
436
5616
684.
2128
268
300.
220.
9200
748.
3300
356.
324.
276.
492.
292
552
240
264.
328.
372
304
388
316.
166.94 F
168.86 F
174.09 F
175.16 F
178.98 F
180.12 F
181.09 F
1 85 1 4 F
186 11 F
187.11 F
192.16 F
193.09 F
198.03 F
199.06 F
204.08 F
20508 F
20608 F
207.12 F
211 05
217.00 F
21797 F
220.98 F
221.97 F
223.06 F
224.06 F
22508 F
227.08 F
22902 F
244.00 F
245 12 F
24605 F
255.03 F
256.06 F
258.16 F
265.09
272.94 F
27403 F
27503 F
27606 F
277.06 F
29600 F
323.06 F
33403 F
364.94 F
372.03
422.97 F
440.97 F
441.97 F
442.97 F
444.03 F
4.99
261
098
1.66
324
2.33
1.33
1 75
11 48
3.26
1.13
1 04
10000
700
2.82
5 12
1986
2.95
2.35
574
1.02
734
1.51
1.60
11.42
2.97
5.39
098
927
1.08
1.64
43.22
667
2.68
1 24
1 90
3.64
1826
2.55
1 64
492
2.13
1.26
2.44
097
4 12
687
49.78
989
0.97
%RIC
062
032
0 12
020
0.40
0.29
0 16
022
1 42
040
0.14
0.13
12.34
086
035
0.63
2.45
0.36
029
071
013
091
0.19
020
1.41
034
067
0.12
1 14
0.13
0.20
533
082
033
0 15
023
0.45
2.25
0.31
020
0.61
0.26
0 16
030
0.12
0.51
085
614
1.22
012
Inten
1096.
572.
216.
364
712.
512.
292
384
2520
716
248
228.
21952.
1536
620
1124
4360
648.
516.
1260
224.
1612.
332.
352.
2508
612.
1184
216
2036
236
360
9488
1464
588.
272.
416
800
4008.
560.
360
1080
468.
276
536
212
904
1508
10928
2172.
212.
G-7
-------
Figure G-1. Base/neutrals standard reconstructed ion chromatogram for 3-day and 7-day PatapscoPOTW base/neutral fraction
effluent analysis.
MIDRIC
06/22/8412:04:00
Sample Semivolatiles Analysis
Conds.: 35 to 290
Range. G 1,5259 Label' N 0, 4.0
Data: BNSTD622 #1
Call. BNSTD622#2
Quan: A 0, 1.0 J 0 Base: U 20, 3
Scans 1 to 5259
1000-1
RIC .
3284990.
1000
7:15
2000
14-31
3000
21.46
4000
29:02
—I
5000
36:17
Scan
Time
G-8
-------
Figure G-2. Surrogate spike standard reconstructed ion chromatogram for 3-day and 7-day Patapsco POTW base/neutral
fraction effluent analysis.
MIDRIC
06/22/84 10-58.00
Sample Semivolatiles Analysis
Conds • 35 to 290
Range G 1,5259 Label: N 0, 4.0
Data: SPKSTD622 #1
Call: SPKSTD622 #2
Quan A 0, 1.0 J 0 Base: U 20, 3
Scans 1 to 5259
100.0-,
RIC
3104760
2000
14:31
3000
21 46
4000
29:02
I
5000
36 17
Scan
Time
G-9
-------
Figure G-3. Blank reconstructed ion chromatogram for 3-day and 7-day PatapscoPOTW base/neutral fraction effluent analysis.
100.0 -i
MIDRIC
06/22/84 16-03:00
Sample Semivolatiles Analysis
Conds. 35 to 290
Range. G 1,5259 Label N 0, 4.0
Data: BNBLK2622 #1
Cah: BNBLK2622 #2
Quan: A 0, 1.0 J 0 Base: U 20, 3
Scans 1 to 5259
RIC .
1548280.
1000
715
2000
14.31
3000
21:46
4000
29:02
5000
36:17
Scan
Time
G-10
-------
Figure G-4. Reconstructed ion chromatogram for 3-day Patapsco POTA/V base/neutral fraction effluent analysis.
MIDRIC
06/23/84 5:20:00
Sample: Semivolatiles Analysis
Conds.: 35 to 290
Range: G 1,5259 Label: N 0, 4.0
Data:TOX1758BN#1
Cali:TOX1755BN#2
Quan: A 0, 1.0 J 0 Base: U 20, 3
Scans 1 to 5259
100.0-1
RIC _
166400.
1380
I.S.
I.S.
IS.
I
1000
7:15
2000
14:31
3000
21 46
4000
29:02
5000
36.17
Scan
Time
G-n
-------
Figure G-5. Library search for possible compound from 3-day Patapsco POTW base/neutral fraction effluent analysis.
MID Library Search
06/23/845 2000+ 1001'
Sample Semivolatiles Analysis
Conds 35 to 290
Data TOX1758BN #1
Cah TOX1758BN#2
Base M/Z 68
RIC 109311.
1214i
Sample
I
C7 H10CI.2
1214]
M Wt 1 64
B Pk68
Rank 1
In 9689
Pur 711
I Jl
ill
I.LlI Ll I LL II,. 11.. . -. .
Bicyclo[4.1 .0]Heptane, 7,7-Dichloro-
I
1
hi III , II .
1214
Sample Minus Library
iJI I.I, ill), lu., nli. l.i III. 4.11..
-1214-
M/Z 50
100
150
200
250
300
350
G-12
-------
Figure G-6. Reconstruction ion chromatogram for 7-day Patapsco base/neutral fraction effluent analysis.
Data TOX1773BN #1
Call TOX1773BN #2
100.0-1
RIC .
MIDRIC
06/23/84 6:07:00
Sample. Semivolatiles Analysis
Conds.' 35 to 290
Range: G 1,5259 Label N 0, 4.0 Quan A 0, 1 0 J 0 Base U 20, 3
I.S
Scans 1 to 5259
111104.
I.S
I.S
2000
14:31
3000
21.46
4000
29:02
5000
36 17
Scan
Time
G-13
-------
Figure G-7. DFTPP reconstructed ion chromatogram for 3-day and 7-day Patapsco POTW base/neutral fraction effluent
analysis.
MIDRIC
06/22/8485000
Sample DFTPP (50 m)
Conds 125 to 220
Range G 1,1 377 Label N 0, 4.0
100.0-,
Data- DFTPP622A#1
Cah DFTPP622A #2
Quan A 0, 1 0 J 0 Base. U 20, 3
1153
Scans 1100 to 1200
RIC .
202752.
1110
1121 11271132
1142
1179
1191
1100
7 59
I
1120
8:00
1 1
1140
8:16
1
1160
8:25
^- —
1
1180
8.34
I
1200
8:43
Scan
Time
G-14
-------
Figure G-8. DFTPP mass spectrum for 3-day and 7-day Patapsco base/neutral POTW fraction effluent analysis.
MID Mass Spectrum
06/22/848:50:00 + 8:22
Sample: DFTPP
Conds.: 125 to 220
GCTemp: 220 Deg. C
Data: DFTPP622A #1
Cah DFTPP622A#2
Base M/Z: 198
Rir 177920
100.0-
50.0-
•
M/Z
100.0-
50.0-
M/Z
4
51
0.1
L
IT=I —
5
69
.1
57.1
.0
II
1 »-•
127.1
11
93.1 I
III! ..I...JI 1 ... li
D 100
D.O
11
1 , ,
7.0
I 18
141.2 159.2, 179°
150
255.0
250
L26f
275.0
i
.1 , 284.8
il I.. _ _ . .
~T ' - r *
— r—
1 31 6.0 i
i:j •. -i -i: r'1
300
334.0
"
5.1
I.. ... 1
.> vy
2061
224.1
I --ill ' I- li'li • Li
~|iz
------- |