FIELD VERIFICATION PROGRAM
(AQUATIC DISPOSAL)
TECHNICAL REPORT D-87-7
EFFECTS OF BLACK ROCK HARBOR DREDGED
MATERIAL ON THE SCOPE FOR GROWTH
OF THE BLUE MUSSEL, MYTILUS EDULIS,
AFTER LABORATORY AND FIELD EXPOSURES
by
William G. Nelson, Donald K. Phelps, Walter B. Galloway,
Peter F. Rogerson, Richard J. Pruell
Environmental Research Laboratory
US Environmental Protection Agency
Narragansett, Rhode Island 02882
September 1987
Final Report
Approved For Public Release, Distribution Unlimited
Prepared for DEPARTMENT OF THE ARMY
US Army Corps of Engineers
Washington, DC 20314-1000
and
US Environmental Protection Agency
Washington, DC 20460
Monitored by Environmental Laboratory
US Army Engineer Waterways Experiment Station
PO Box 631, Vicksburg, Mississippi 39180-0631
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Destroy this report when no longer needed Do not return
it to the originator.
The findings in this report are not to be construed as an official
Department of the Army position unless so designated
by other authorized documents.
The contents of this report are not to be used for
advertising, publication, or promotional purposes.
Citation of trade names does not constitute an
official endorsement or approval of the use of
such commercial products.
The D-series of reports includes publications of the
Environmental Effects of Dredging Programs:
Dredging Operations Technical Support
Long-Term Effects of Dredging Operations
Interagency Field Verification of Methodologies for
Evaluating Dredged Material Disposal Alternatives
(Field Verification Program)
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SUBJECT: Transmittal of Field Verification Program Technical Report Entitled
Effects of Black Robk Harbor Dredged Material on the Scope for
Growth of the Blue Mussel, Mytitus Edulis, after Laboratory and
Field Exposures"
TO: All Report Recipients
1. This is one in a series of scientific reports documenting the findings of
studies conducted under the Interagency Field Verification of Testing and Pre-
dictive Methodologies for Dredged Material Disposal Alternatives (referred to
as the Field Verification Program or FVP). This program is a comprehensive
evaluation of environmental effects of dredged material disposal under condi-
tions of upland and aquatic disposal and wetland creation.
2. The FVP originated out of the mutual need of both the Corps of Engineers
(Corps) and the Environmental Protection Agency (EPA) to continually improve
the technical basis for carrying out their shared regulatory missions. The
program is an expansion of studies proposed by EPA to the US Army Engineer
Division, New England (NED), in support of its regulatory and dredging mis-
sions related to dredged material disposal into Long Island Sound. Discus-
sions among the Corps' Waterways Experiment Station (WES), NED, and the EPA
Environmental Research Laboratory (ERLN) in Narragansett, RI, made it clear
that a dredging project at Black Rock Harbor in Bridgeport, CT, presented a
unique opportunity for simultaneous evaluation of aquatic disposal, upland
disposal, and wetland creation using the same dredged material. Evaluations
were to be based on technology existing within the two agencies or developed
during the six-year life of the program.
3. The program is generic in nature and will provide techniques and interpre-
tive approaches applicable to evaluation of many dredging and disposal opera-
tions. Consequently, while the studies will provide detailed site-specific
information on disposal of material dredged from Black Rock Harbor, they will
also have great national significance for the Corps and EPA.
4. The FVP is designed to meet both Agencies' needs to document the effects
of disposal under various conditions, provide verification of the predictive
accuracy of evaluative techniques now in use, and provide a basis for deter-
mining the degree to which biological response is correlated with bioaccumula-
tion of key contaminants in the species under study. The latter is an
important aid in interpreting potential biological consequences of bioaccumu-
lation. The program also meets EPA mission needs by providing an opportunity
to document the application of the generic predictive hazard-assessment
research strategy applicable to all wastes disposed in the aquatic environ-
ment. Therefore, the ERLN initiated exposure-assessment studies at the
aquatic disposal site. The Corps-sponsored studies on environmental conse-
quences of aquatic disposal will provide the effects assessment necessary to
complement the EPA-sponsored exposure assessment, thereby allowing ERLN to
develop and apply a hazard-assessment strategy. While not part of the Corps-
funded FVP, the EPA exposure-assessment studies will complement the Corps'
work, and together the Corps and the EPA studies will satisfy the needs of
both agencies.
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SUBJECT: Transmittal of Field Verification Program Technical Report Entitled
"Effects of Black Robk Harbor Dredged Material on the Scope for
Growth of the Blue Mussel, Mytilus Edulis, after Laboratory and
Field Exposures"
5. In recognition of the potential national significance, the Office, Chief
of Engineers, approved and funded the studies in January 1982. The work is
managed through the Environmental Laboratory's Environmental Effects of Dredg-
ing Programs at WES. Studies of the effects of upland disposal and wetland
creation were conducted by WES, and studies of aquatic disposal were carried
out by the ERLN, applying techniques worked out at the laboratory for evaluat-
ing sublethal effects of contaminants on aquatic organisms. These studies
were funded by the Corps while salary, support facilities, etc., were provided
by EPA. The EPA funding to support the exposure-assessment studies followed
in 1983; the exposure-assessment studies are managed and conducted by ERLN.
6. The Corps and EPA are pleased at the opportunity to conduct cooperative
research and believe that the value in practical implementation and improve-
ment of environmental regulations of dredged material disposal will be consid-
erable. The studies conducted under this program are scientific in nature and
are published in the scientific literature as appropriate and in a series of
Corps technical reports. The EPA will publish findings of the exposure-
assessment studies in the scientific literature and in EPA report series. The
FVP will provide the scientific basis upon which regulatory recommendations
will be made and upon which changes in regulatory implementation, and perhaps
regulations themselves, will be based. However, the documents produced by the
program do not in themselves constitute regulatory guidance from either
agency. Regulatory guidance will be provided under separate authority after
appropriate technical and administrative assessment of the overall findings of
the entire program.
_ yv I j,
'"-Jane® Choromokos, Jr., Pn.D., P.E.
Director, Research and Development
U. S. Army Corps of Engineers
Bernard D. Goldstein, M.D.
Assistant Administrator for
Research and Development
U. S. Environmental Protection
Agency
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SECURITY CLA^SI^CAtIoN OF THIS PAGE
REPORT DOCUMENTATION PAGE
Form Approved
QMS Ho 0704 0188
C'P Pitt tun 30. 1986
1a REPORT SECURITY CLASSIFICATION
IlnrlflRRiflpa
lb RESTRICTIVE MARKINGS
2a SECURITY CLASSIFICATION AUTHORITY
2b DECLASSIFICATION/DOWNGRADING SCHEDULE
3 DISTRIBUTION /AVAILABILITY OF REPORT
Approved for public release; distribution
unlimited.
4 PERFORMING ORGANIZATION REPORT NUMBER(S)
5. MONITORING ORGANIZATION REPORT NUMBER(S)
Technical Report D-87-7
6a NAME OF PERFORMING ORGANIZATION
USEPA, Environmental Research
Laboratory
6b. OFFICE SYMBOL
(If spplictble)
7a NAME OF MONITORING ORGANIZATION
USAEWES
Environmental Laboratorv
6c. ADDRESS (Oty. Stat#. and ZIP Cod*)
Narragansett, RI 02882
7b. ADDRESS (City, Stat*, tnd ZIP Code)
PO Box 631
Vicksburg, MS 39180-0631
8*. NAME OF FUNDING /SPONSORING
ORGANIZATION
See reverse.
8b OFFICE SYMBOL
(If applicable)
9. PROCUREMENT INSTRUMENT IDENTIFICATION NUMBER
8t. ADORESS (City, Staff, and ZIPCodt)
Washington, DC 20314-1000;
j
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Unclassified
IICUMTY CLASSIFICATION OF THIS *A««
8a. NAME OF FUNDING/SPONSORING ORGANIZATION (Continued).
US Army Corps of Engineers;
US Environmental Protection Agency
12. PERSONAL AUTHOR(S) (Continued).
Nelson, William G.; Phelps, Donald K.; Galloway, Walter B.; Rogerson, Peter F.;
Pruell, Richard J.
18. SUBJECT TERMS (Continued).
Dredged material (WES)
Dredging—Connecticut—
Black Rock Harbor (LC)
Dredging—Environmental aspects (LC)
Marine pollution (LC)
Mytilue edulis (WES)
Mussels—Environmental aspects (LC)
19. ABSTRACT (Continued).
In the field, mussels were placed along a transect from the center of the disposal
mound to a clean area distant from the disposal mound. Exposure estimates indicated that
the maximum concentration of BRH material occurred during the disposal operation, after
which both exposure and tissue residue concentrations decreased dramatically. Of the mea-
surements made at the four field stations during the course of the study, a reduction in
the scope for growth of mussels, attributable to BRH material, was observed only once.
The estimated concentration of BRH suspended material (0.7 to 0.2 mg/i) during that col-
lection, 8 weeks postdisposal, was very close to the lowest concentration affecting the
scope for growth in the laboratory experiments (1.5 mg/J.).
Uncl«g«< f-Igrl
ICCUMITV C LA Ml f I CATION OW THIS PAQC
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PREFACE
This report describes work performed by the US Environmental Protection
Agency (EPA), Environmental Research Laboratory, Narragansett, R. I. (ERLN),
as part of the Interagency Field Verification of Testing and Predictive
Methodologies for Dredged Material Disposal Alternatives Program (Field Veri-
fication Program (FVP)). The FVP was sponsored by the Office, Chief of Engi-
neers (OCE) , and was assigned to the US Army Engineer Waterways Experiment
Station (WES), Vicksburg, Miss. The objective of this interagency program was
to field verify existing predictive techniques for evaluating the environmen-
tal consequences of dredged material disposal under aquatic, intertidal, and
upland conditions. The aquatic portion of the FVP was conducted by ERLN, with
the wetland and upland portions conducted by WES.
The principal investigators for this aquatic study and authors of this
report were Mr. William G. Nelson and Dr Richard J. Pruell of Science Applica-
tions International Corporation (SAIC) and Dr. Donald K. Phelps, Mr. Walter B.
Galloway, and Dr. Peter F. Rogerson of ERLN. Laboratory-cultured algae were
provided by Mr. Gregory Tracey, SAIC. Technical support for the scope for
growth measurements was provided by Mr. William Giles, ERLN. Diving support
for the field portion of the study was provided by Messrs. Bruce Reynolds and
Norman Rubenstein, ERLN, and Mr. Tracey, SAIC.
Analytical chemistry support was provided by Dr. Gerald Hoffman,
Mr. Richard Lapan, Mr. Curtis Norwood, and Mr. Frank Osterman, ERLN;
Mr. Richard McKinney, Mr. Warren Boothman, Ms. Adria Elskus, Ms. Eileen
McFadden, Mr. Lawrence LeBlanc, Mr. Robert Bowen, and Ms. Sharon Pavignano,
SAIC; and Ms. Kathleen Schweitzer, University of Rhode Island.
Mses. Joan E. Seites, Barbara G. Gardner, and Colette J. Brown, Computer
Sciences Corporation (CSC), provided word processing support in preparation of
the draft report. In addition, assistance in statistical analysis was pro-
vided by Dr. James Heltshe, CSC, and Dr. Clifford H. Katz, SAIC. Critical
reviews of this report were provided by Dr. Katz, SAIC, and Drs. John H.
Gentile and Gerald G. Pesch, ERLN. Technical reviews by WES personnel were
also provided,
The OCE Technical Monitors were Drs. John Hall, Robert J. Pierce, and
William L. Klesch. The EPA Technical Director for the FVP was
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Dr. John Gentile; Technical Coordinators were Mr. Walter Galloway and
Dr. Gerald Pesch; and Project Manager was Mr. Allan Beck.
The study was conducted under the direct WES management of
Drs. Thomas M. Dillon and Richard Peddicord and under the general management
of Dr. C. Richard Lee, Chief, Contaminant Mobility and Criteria Group;
Mr. Donald L. Robey, Chief, Ecosystem Research and Simulation Division; and
Dr. John Harrison, Chief, Environmental Laboratory. The Environmental Effects
of Dredging Programs Manager was Dr. Robert M. Engler, with Mr. Robert L.
Lazor, FVP Coordinator. Dr. Thomas D. Wright was the WES Technical Coordina-
tor for the FVP reports. The report was edited by Ms. Jessica S. Ruff of the
WES Information Products Division.
COL Dwayne G. Lee, CE, was the Commander and Director of WES.
Dr. Robert W. Whalin was Technical Director.
This report should be cited as follows:
Nelson, W. G., et al. 1987. "Effects of Black Rock Harbor
Dredged Material on the Scope for Growth of the Blue Mussel,
Mytilus edulis, After Laboratory and Field Exposure," Technical
Report D-87-7, prepared by the US Environmental Protection Agency,
Narragansett, R. I., for the US Army Engineer Waterways Experiment
Station, Vicksburg, Miss.
2
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CONTENTS
Page
PREFACE 1
LIST OF TABLES 4
LIST OF FIGURES 4
PART I: INTRODUCTION 7
Background 7
Project Description 9
Project Scope 11
Laboratory-Field Comparisons 12
Residue-Effects Relationships 12
Scope for Growth 13
PART II: MATERIALS AND METHODS 15
Chemical Methods 15
Laboratory Methods 19
Field Methods 28
PART III: RESULTS 32
Laboratory 32
Field 61
Laboratory-to-Field Comparison 82
PART IV: DISCUSSION 84
Laboratory Experiments 84
Field Experiments 88
Laboratory-to-Field Comparison 93
PART V: CONCLUSIONS 96
REFERENCES 97
APPENDIX A: CHEMICAL FORMULAS AND FIELD MUSSEL RESIDUE
CONCENTRATIONS A1
APPENDIX B: COMPARISON OF SELECTED CONTAMINANTS IN
BRH AND REF SEDIMENTS B1
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LIST OF TABLES
No. Page
1 Collection Information for the Mussels Used in the
Laboratory Experiments 19
2 Cruise Number, Deployment Date, Retrieval Date, and
Length of Deployment for Mussels Transplanted to CLIS 29
3 Suspended Sediment Concentrations in the Mussel
Exposure System 32
4 Chemical Monitoring of the Exposure System in Experiment 2 34
5 PCB Tissue Residues in Mussels from Laboratory Experiment 1 35
6 PCB Tissue Residues in Mussels from Laboratory Experiment 2 35
7 PCB Concentrations in Mussels from Both Laboratory
Experiments 37
8 Clearance Rates of Mussels in Dosing System Exposure Tanks 45
9 Clearance Rates of Mussels from the Two Laboratory Exposures.... 46
10 Absorption Efficiencies of Mussels from the Two
Laboratory Exposures 47
11 Respiration Rates of Mussels from the Two
Laboratory Exposures 47
12 Ammonia Excretion Rates of Mussels from the Two
Laboratory Exposures 48
13 SFG Values of Mussels from the Two Laboratory Exposures 49
14 Actual Growth of Mussels in the Exposure Chambers 50
15 Predicted BRH Suspended Material Sediment Exposure Required
to Achieve the Measured Tissue Residue Values of Mussels
Deployed in CLIS 62
16 Predicted BRH Suspended Sediment Exposure Based on PCB and
Copper Whole Water Chemistry Data 65
17 Mean Values of the Physiological Parameters Measured on
Mussels from CLIS Collections with the Mean SFG Value
Calculated for Station 73
LIST OF FIGURES
No. Page
1 Central Long Island Sound disposal site and Black Rock Harbor
dredge site 9
2 FVP sampling stations 10
3 Suspended sediment dosing system 20
4 Suspended sediment oxidation system.................... 22
5 Laboratory exposure system 23
6 Concentrations of PCB as A1254 in the tissue of M. edulis
exposed to BRH suspended sediments for 28 days 36
7 Concentrations of PCB as A1254, normalized for lipids, in the
tissue of M. edulis exposed to BRH sediment for 14 days 37
8 Concentrations of phenanthrene and 178 alkyl homologs in the
tissue of M. edulis esposed to BRH suspended sediments for
28 days 39
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LIST OF FIGURES (Continued)
No. Page
9 Concentrations of fluoranthene and benzo(a)pyrene in the tissue
of M. edulis exposed to BRH suspended sediments for 28 days... 40
10 Concentrations of the SUM of PAHs and CENT of PAHs in the tissue
of M. edulis exposed to BRH suspended sediments for 28 days... 41
11 Concentrations of ethylan and PCB as A1254 in the tissue of
M, edulis exposed to BRH suspended sediments for 28 days 42
12 Concentrations of cadmium and copper in the tissue of M. edulis
exposed to BRH suspended sediments for 28 days 43
13 Concentrations of chromium and iron in the tissue of M. edulis
exposed to BRH suspended sediments for 28 days 44
14 Relationship between the SFG of M. edulis and BRH exposure
concentration on Days 14 and 28 of the laboratory
experiments 50
15 Effect of BRH suspended sediment on the clearance rate, SFG,
and shell growth of M. edulis, Day 14, Experiment 1 51
16 Effect of BRH suspended sediment on the clearance rate, SFG,
and shell growth of M. edulis, Day 14, Experiment 2 52
17 Effect of BRH suspended sediment on the clearance rate, SFG,
and shell growth of M. edulis, Day 28, Experiment 2 53
18 Relationship between the SFG of M. edulis and the tissue
residue concentrations of phenanthrene and the sum of the
178 alkyl homologs in the laboratory experiments 55
19 Relationship between the SFG of M. edulis and the tissue
residue concentrations of fluoranthene and benzo(a)pyrene in
the laboratory experiments 56
20 Relationship between the SFG of M. edulis and the summary
statistics, SUM and CENT, in the laboratory experiments 57
21 Relationship between the SFG of M. edulis and the tissue
residue concentrations of PCB as A1254 and ethylan in the
laboratory experiments 58
22 Relationship between the SFG of M. edulis and the tissue
residue concentrations of cadmium and copper in the
laboratory experiments 59
23 Relationship between the SFG of M. edulis and the tissue
residue concentrations of chromium and iron in the
laboratory experiments 60
24 Concentrations of phenanthrene and the 178 alkyl homologs in
the tissues of M. edulis exposed at the specified FVP sta-
tions and sampling dates 67
25 Concentrations of fluoranthene and benzo(a)pyrene in the
tissues of M. edulis exposed at the specified FVP stations
and sampling dates 68
26 Concentrations of the sum of the PAHs and CENT in the tissues
of M. edulis exposed at the specified FVP stations and
sampling dates 69
27 Concentrations of PCBs as A1254 and ethylan in the tissues of
M. edulis exposed at the specified FVP stations and sampling
dates 70
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LIST OF FIGURES (Continued)
No. Page
28 Concentrations of cadmium and copper in the tissues of M. edulis
exposed at the specified FVP stations and sampling dates 71
29 Concentrations of chromium and iron in the tissues of M. edulis
exposed at the specified FVP stations and sampling dates 72
30 Relationship between the SFG of M. edulis and PCB tissue residue
concentrations in laboratory and field-exposed animals 78
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EFFECTS OF BLACK ROCK HARBOR DREDGED MATERIAL ON THE SCOPE
FOR GROWTH OF THE BLUE MUSSEL, MYTILUS EPULIS,
AFTER LABORATORY AND FIELD EXPOSURES
PART I: INTRODUCTION
Background
1. The Marine Protection, Research, and Sanctuaries Act (Public
Law 92-532) was passed by Congress in 1972. This law states that it is the
policy of the United States to regulate disposal of all types of materials
into ocean waters and to prevent or strictly limit disposal of any material
that would adversely affect human health, welfare, the marine environment, or
ecological systems. The implementation of this law, through the issuance of
permits as defined in the final regulations and criteria, is shared jointly by
the US Environmental Protection Agency (USEPA) and the US Army Corps of
Engineers (CE).
2. In 1977, the CE and the USEPA prepared technical guidance for the
implementation of the final ocean dumping regulations in the form of a manual
entitled "Ecological Evaluation of Proposed Discharge of Dredged Material into
Ocean Waters" (USEPA/CE 1977). This manual specified which test procedures
were to be followed in collecting information to be used in making a disposal
decision. Among the procedures were those for: (a) chemically characterizing
the proposed dredged material; (b) determining the acute toxicity of liquid,
suspended particulate, and solid phases; (c) estimating the potential
contaminant bioaccumulation; and (d) describing the initial mixing during
disposal. These methods have been used for determining the suitability of
dredged material for open-water disposal. The procedures in this manual rep-
resented the technical state of the art at that time and were never intended
to be inflexible methodologies. The recommended test methods were chosen to
provide technical information that was consistent with the criteria specified
in the regulations. However, use of the manual in the permit process has
identified conceptual and technical limitations with the recommended test
methods (Gentile and Scott 1986).
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3. To meet this critical need, the Interagency Field Verification of
Testing and Predictive Methodologies for Dredged Material Disposal Alterna-
tives Program or the Field Verification Program (FVP) was authorized in 1982.
This 6-year program was sponsored by the Office, Chief of Engineers, and was
assigned to the US Army Engineer Waterways Experiment Station (WES), Vicks-
burg, Miss. The objective of this interagency program was to field verify
existing test methodologies for predicting the environmental consequences of
dredged material disposal under aquatic, intertidal, and upland conditions.
The aquatic portion of the FVP was conducted by the USEPA Environmental Re-
search Laboratory, Narragansett, R. I. (ERLN). The wetland and upland por-
tions, conducted by WES, are reported in separate documentation.
4. The USEPA ERLN was responsible for conducting research on the aqua-
tic option for disposal of dredged material. There were three research objec-
tives for this portion of the program. The first was to demonstrate the
applicability of existing test methods to detect and measure effects of
dredged material, and to determine the degree of variability and reproducibil-
ity inherent in the testing procedure. This phase of the program (Laboratory
Documentation) is complete and the results are published in a series of tech-
nical reports. This information provides insight into how the various methods
function, their sources of variability, their respective and relative sensi-
tivities to the specific dredged material being tested, and the degree of con-
fidence that can be placed on the data derived from the application of the
methods.
5. The second objective was to field verify the laboratory responses by
measuring the same response under both laboratory and field exposures. A
basic and often implicit assumption is that results derived from laboratory
test methods are directly applicable in the field. While this assumption is
intuitive, there are no supporting data from studies on complex wastes in the
marine environment. The study reported herein offers a unique opportunity to
test this basic assumption.
6. The third objective was to determine the degree of correlation of
tissue residues resulting from bioaccumulation of dredged material contami-
nants with biological responses from laboratory and field exposure to dredged
material. However, this study was not designed to address cause-effect rela-
tionships, and the multicontaminant nature of the dredged material precludes
any such assumptions.
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Project Description
7. The aquatic disposal portion of the FVP was a site- and waste-
specific case study that applied the concepts and principles of risk assess-
ment. The disposal site for the FVP was an historical site known as the
Central Long Island Sound (CLIS) disposal site (1.8 by 3.7 km) located approx-
imately 15 km southeast of New Haven, Conn. (Figure 1). The sedimentology at
Figure 1. Central Long Island Sound disposal site
and Black Rock Harbor dredge site
the disposal and reference sites is primarily silt-clay, with a mean grain
size of 0.013 mm. Thermal stratification occurs from April to September, and
during this period bottom salinity is slightly higher than that of the sur-
face. Tidal currents typically dominate the near-bottom water in an east-west
direction. The net bottom drift is to the northwest at 0.5 cm/sec. Suspended
sediment concentrations average 10 mg/Jl, with storm-induced values to 30 mg/£.
The baseline community data revealed a homogeneous, mature infaunal community
dominated by the polychaete Nephtys inaisa and the bivalve molluscs Nuoula
proxima and Yoldia limatula.
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8. The FVP disposal site was selected within the CLIS so as to minimize
contamination from other sources, including relic disposal operations or on-
going disposal activities occurring during the study period. This was neces-
sary to ensure a point source of contamination. The uniformity of physical,
chemical, and biological properties of the disposal site prior to disposal
allowed detection of changes-in these properties due to the disposal of the
dredged material. Finally, the stations used to study the biological effects
in this study were selected along the primary axis of current flow to repre-
sent a gradient of potential exposure for the biota (Figure 2).
72
41 05
52.5 72
52.0 72
FVP DISPOSAL
U"" ^ •N
51.5 72
SITE
i\
51.0
41 0<
/
/
/
/
1
\
\
s
r—i r—r" i ¦ ¦»
0 500m
>.0
^
/ //' N)
ii, •
\ \\ CNTFf 20<
* '
"
-
REFS
3 km
*\'x
|}\ \
i •
3E 400E
/ /
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1
Figure 2. FVP sampling stations
9. The spatial scale of this study was near-field and limited to the
immediate vicinity of the disposal site. A primary assumption was that the
mound of dredged material constitutes a point source of contamination. The
temporal scale for the study was 4 years, which included a year of predisposal
data collection to define seasonal patterns in the physical, chemical, and bi-
ological variables and 3 years of postdisposal data collection to address the
objectives of the program and to evaluate the long-term impacts of the dis-
posal operation on the surrounding benthic communities.
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10. The dredging site was Black Rock Harbor (BRH), located in
Bridgeport, Conn., where maintenance dredging provided a channel 46 m wide and
3
5.2 m deep at mean low water (Figure 1). Approximately 55,000 m of material
were dredged during April and May 1983 and disposed in 20 m of water in the
northeastern corner of the CLIS disposal site.
11. The dredged material from BRH contained substantial concentrations
of both organic and inorganic contaminants (Rogerson, Schimmel, and Hoffman
1985) . Polychlorinated biphenyls (PCBs) were present in the dredged material
at a concentration of 6,800 ng/g, and polynuclear aromatic hydrocarbons (PAHs)
with molecular weights between 166 and 302 were present at concentrations
ranging from 1,000 to 12,000 ng/g. Alkyl homologs of the PAHs were also pre-
sent in the dredged material at concentrations between 1,000 and 13,000 ng/g.
Inorganic contaminants of toxicological importance present in the dredged
material include copper (2,380 yg/g), chromium (1,430 yg/g), zinc
(1,200 yg/g), lead (380 yg/g), nickel (140 yg/g), cadmium (23 yg/g), and
mercury (1.7 yg/g).
Project Scope
12. The FVP was unique among marine research studies for several rea-
sons. The program objectives were directly focused on addressing specific
limitations in the methodologies and interpretive framework of the current
regulatory process. Among the program strengths were: the development and
evaluation of a suite of biological endpoints that used the same material; the
biological tests represented different levels of biological organization; the
tests were conducted under both laboratory and field exposure conditions; tis-
sue residues were examined concurrently with measurements of biological ef-
fects; the duration of the study was adequate to evaluate the use of community
responses as a benchmark against which other biological responses could be
compared; and the project was a site- and waste-specific case study for the
application and evaluation of the components of a risk assessment, including
the development of methodologies for predicting and measuring field exposures
in the water column and benthic compartments. Limitations of this study were:
only one dredged material was evaluated, which constrained certain types of
comparisons; the size of the study put limits on the extent to which any given
objective could be examined; and the resources allocated to determine field
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exposures were insufficient. The last constraint was particularly important
because the laboratory-field comparisons and the risk assessment process both
required accurate predictions of environmental exposures.
Laboratory-Field Comparisons
13. The field verification of laboratory test methods was designed to
compare the exposure-response relationships measured in both the laboratory
and the field. Exposure for the purposes of this discussion includes the
total dredged material with all of its contaminants. Specific contaminants
are used as "tracers" to verify the exposure environment, which is described
in terms of BRH dredged material, and to illustrate exposure-response rela-
tionships between the laboratory and field. The specific contaminants are a
subset of a comprehensive suite of chemicals analyzed in this study and were
selected based upon their environmental chemistry and statistical representa-
tiveness. The use of specific contaminants in no way implies a cause-and-
effect relationship between contaminant and response.
14. Exposure in open marine systems is characterized by highly dynamic
temporal and spatial conditions and cannot be completely replicated in labo-
ratory systems. Consequently, the approach chosen for this program was to
develop laboratory exposure-response data using only general field exposure
information. The most appropriate statistical analyses for laboratory-field
comparisons are observational where all the variables from the laboratory and
field are used to identify similarities and differences, independent of any
limiting assumptions.
Residue-Effects Relationships
15. Determining the relationship between contaminant tissue residues
resulting from bioaccumulation and the biological responses measured is a
principal objective of this program. Such relationships do not in any way
imply cause and effect, but rather seek to determine the statistical relation-
ship between an effect and any associated residues. The approach used is to
determine specific contaminant residues in the tissues of the organisms as the
result of exposure to the whole dredged material in both the laboratory and
the field. These residues are determined at the same time that biological
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responses are being measured. Residue-effect relationships will be described
and interpreted for both laboratory and field exposures.
Scope for Growth
16. The scope for growth (SFG) index (Warren and Davis 1967) is a mea-
sure of the energy available to an organism for production, both somatic and
reproductive, after accounting for routine metabolic costs. The SFG value
represents the instantaneous assessment of energy balance in an organism for
that set of environmental conditions under which it is measured.
17. A very important point pertinent to the interpretation of SFG
measurements is that this index may be used to test several hypotheses. The
present study investigated the hypothesis that exposure to different environ-
mental conditions (concentrations of BRH dredged material) may result in last-
ing physiological effects in an organism, even after its removal from those
exposure conditions. Consistent with this hypothesis, the SFG of mussels was
measured under standardized conditions, after separate laboratory and field
exposures. These measurements were made to test for relative differences be-
tween stations (field) or between exposure concentrations (laboratory). SFG
differences in this instance were interpreted as being caused by the respec-
tive environmental conditions to which the organisms were exposed. This is
true since, under standardized conditions, mussels of similar physiological
condition should exhibit similar SFG responses. This approach was used be-
cause one of the goals of this study was to compare the laboratory and field
SFG results. Measurement of SFG under separate laboratory and field condi-
tions would not allow this comparison.
18. There are sufficient historical data to indicate that the use of
the SFG index with the blue mussel, Mytilus edulis, might be useful in assess-
ing the biological impact of disposed dredged material in the marine environ-
ment. The SFG index proved useful as a response parameter for measuring
physiological effects of BRH dredged material on M. edulis in the laboratory
(Nelson, Black, and Phelps 1985). Stickle et al. (1985) reported an inverse
relationship between the SFG of mussels and exposure to increasing concentra-
tions of the water-soluble fraction of crude oil. Widdows et al. (1982) also
reported a dose-response effect between SFG and aromatic petroleum hydrocarbon
exposure concentrations in mussels. In the field, Widdows, Phelps, and
13
-------
Galloway (1981) reported a decrease in the SFG of mussels in response to in-
creasing levels of pollution in Narragansett Bay.
19. The objectives of this portion of the FVP with respect to the SFG
index were to: (a) determine whether there was a relationship between contam-
inant residues in tissues and subsequent biological effects measured in both
the laboratory and the field, and (b) determine whether the responses measured
in the laboratory exposures were comparable to the responses measured in field
organisms for similar exposures. The evaluation of these objectives will form
the basis of this field verification report.
14
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PART II: MATERIALS AND METHODS
Chemical Methods
Analytical methods
20. The analytical methods used in this study are presented here in
summary form. More detailed descriptions of the analytical methods are avail-
able in Lake, Hoffman, and Schimmel (1985). Most of these methods represent
extensive modifications of USEPA standard methods developed for freshwater and
wastewater samples. It was necessary to modify these methods to analyze the
types of matrices in this study. These methods were intercalibrated to ensure
the quality of the data.
Organic sample preparation
21. Samples of sediment, suspended particulates, and organisms were
extracted by multiple additions of increasingly less polar organic solvents
using a tissue homogenizer. These mixtures were separated by centrifugation
between additions; polar solvents were removed by partitioning against water;
and the extracts were desulfured with activated copper powder when required.
The extracts were then passed through a precolumn containing activated silica
gel. Samples of both filtered and unfiltered seawater were solvent extracted
in separatory funnels and the extracts saved. Foam plugs containing the dis-
solved organic contaminants from water samples were extracted with organic
solvents. All of the above extracts were subjected to column chromatography
on deactivated silica gel to separate analytical fractions and were volume
reduced carefully prior to analysis.
Organic analysis
22. Electron capture gas chromatographic analyses for PCBs were con-
ducted on a Hewlett-Packard 5840 gas chromatograph equipped with a 30-m DB-5
fused silica column. Samples were quantified against an Aroclor 1254 (A1254)
standard because the distribution of PCB congeners in the dredged material
closely matched that distribution, as did the distribution in organisms at
steady state.
23. Gas chromatograph/mass spectrometric analyses were conducted with a
Finnigan Model 4500 also equipped with a 30-m DB-5 fused silica capillary col-
umn. The mass spectrometer was operated through a standard Incos data system
and was tuned at all times to meet USEPA quality assurance specifications.
15
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24. All instruments were calibrated daily with the appropriate stan-
dards. The concentrations of the standards used were chosen to approximate
those of the contaminants of interest, and periodic linearity checks were made
to ensure the proper performance of each system. When standards were not
available, response factors were calculated using mean responses of comparable
standards. Blanks were carried through the procedure with each set of sam-
ples, and reference tissue homogenate was analyzed with every 12 to 15 tissue
samples.
Organic data compression
25. As stated above, PCBs were quantified as A1254 because the sample
patterns closely resembled that profile. This allowed a convenient way of
reporting these data without treating the voluminous data that would have
resulted from measuring some 55 congener peaks by electron capture detector.
Likewise, a method was sought to summarize the PAH data. Appendix A lists the
35 individual PAH parent and alkyl homolog compounds and groups of compounds
measured in this study. Although useful, this only reduced the data to 14 PAH
variables, which was not sufficient. Since the distribution of PAHs differed
greatly in both quantity and distribution between Long Island Sound and the
BRH dredged material, statistics were sought which would retain significant
quantitative and qualitative information. The quantitative statistic chosen
was the simple SUM of all measured PAHs, and a qualitative descriptor was
chosen by analogy with the center of mass concept from elementary physics and
called a centroid (CENT):
SUM = Z[C(i)] (1)
CENT = E[C(i) * MW(i)]
LtNi SUM <2)
where
til
C(i) = concentration of i PAH from molecular weight 166
through 302, including both parent and alkyl homologs
MW(i) = molecular weight of ith PAH from 166 through 302, including
both parent and alkyl homologs
In this case, CENT describes the "center of mass" of the PAH distribution, and
is in units of molecular weight. It is the concentration-weighted average mo-
lecular weight of any particular PAH distribution. Using this statistic, one
is able to readily distinguish two different sources of PAH distributions, one
16
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with predominately heavy molecular weight pyrogenic compounds, and one with
more lighter molecular weight petrogenic compounds. These distributions are
typically found at the reference station in Long Island Sound (REFS) and BRH,
respectively. A major value to this statistic is that it enables one to
readily distinguish these two sources when their concentrations are nearly
equal. The formulas for calculating these, and 178 alkyl homologs, are shown
in Appendix A. Because distributions of both parents and homologs were mea-
sured, SUMs and CENTs of both parents and homologs were calculated as well.
These were defined as PSUM, PCENT, HSUM, and HCENT. By definition,
SUM = PSUM + HSUM (3)
and
_ (PSUM * PCENT + HSUM * HCENT) ,/N
OhNi - gUM w
It should be noted that dibenzothiophene and its alkyl homologs are not in-
cluded in these calculations because they are not PAHs.
Inorganic sample preparation
26. Sediment was prepared for inorganic analysis by elution at room
temperature with 2N HNO^. The samples were filtered through Whatman
No. 2 filter paper. Organisms were totally digested in concentrated HNO^ at
60° C and filtered through Whatman No. 2 filter paper.
27. Cadmium and copper were concentrated and separated from both the
unfiltered and filtered seawater fractions by coprecipitation (Boyle and
Edmond 1975). The remaining metals (chromium, iron, manganese, nickel, lead,
and zinc) were analyzed by heated graphite atomization atomic absorption
(HGA-AA) via direct injection. Samples of suspended particulates on
Nucleopore (0.45-p) filters were eluted with 2N HNO^ and analyzed by HGA-AA.
Inorganic analysis
28. All flame atomization atomic absorption (FA-AA) was conducted with
a Perkin-Elmer (Model 5000) atomic absorption spectrophotometer. All HGA-AA
determinations were conducted with Perkin-Elmer Model 5000 or 2100 HGA units
coupled to Perkin-Elmer Model 5000 or 603 atomic absorption instruments, re-
spectively. The Model 5000 AA was retrofitted with a Zeeman HGA background
correction unit, and the Model 603 was equipped with a D2 arc background cor-
rection system.
17
-------
29. The FA-AA and HGA-AA instrument operating conditions are similar to
those described in "Methods for Chemical Analysis of Water and Wastes" (USEPA
1979) and those in the manufacturers' reference manuals. The AA instruments
were calibrated each time samples were analyzed for a given element. Sample
extracts were analyzed a minimum of twice to determine signal reproducibility.
Quality assurance checks, conducted after every 15 samples, were analyzed by
the method of standard addition and by analyzing one procedural blank.
Contaminant selection
30. Chemical analyses performed in this study characterize the organic
and inorganic constituents in the dredged material; provide information on the
laboratory and field exposure environments; provide insight into the processes
governing contaminant movement within and between environmental compartments;
and determine which contaminants were accumulated by organisms. Historically,
bulk sediment analyses have been used to characterize dredged material. More
recently, dredged material must be analyzed for USEPA's priority pollutants to
determine if hazardous substances are present and, if so, in what concentra-
tions. While both of these approaches were used in this study, neither ad-
dresses the issue of bioavailability and the potential for contaminants to
bioaccumulate. In this study, bioavailability was determined by examining the
types and distributions of contaminants that bioaccumulated in laboratory
studies (Rogerson, Schimmel, and Hoffman 1985). Based upon the contaminant
profile for the dredged material and residue data, the contaminants selected
for detailed analyses throughout the study included PCBs, PAHs, the pesticide
ethylan, and four metals.
31. A representative subset of chemicals was selected for discussion
throughout the study. The criteria used in selecting this subset included
chemical properties, contaminant representativeness and behavior in various
compartments, and statistical analyses of the distributions of the complete
suite of chemicals analyzed in the program.
32. Multivariate clustering analyses were performed on the chemical
data to define groups or clusters of chemicals that behaved in a statistically
similar manner. No assumptions were made concerning the behavior, inter-
actions, or dynamics of chemicals between compartments; therefore, each com-
partment was analyzed separately. Five compartments were identified from
field and laboratory data for statistical analysis. Of these, the surficial
sediments and the unfiltered, particulate, and dissolved water column
18
-------
fractions described exposure conditions experienced by infaunal and pelagic
organisms. The remaining compartment consisted of tissue residues in
M. edulis.
33. The data were further partitioned into inorganic and organic analy-
ses. The inorganic analyses generally consisted of 8 variables while the or-
ganic analyses contained 61 variables. The clusters of chemicals identified
through the statistical analyses agreed well with those contaminants selected
based on chemical properties and environmental behavior.
Laboratory Methods
Sediment collection
34. Two sediment types were used to conduct laboratory tests for the
field verification studies. The reference sediment (REF) was collected from
the South Reference site (REFS) in Long Island Sound (40°7.95' N and
2
72°52.7' W) by Smith-Maclntyre grab (0.1 m ), press sieved through a 2-mm
sieve, and stored in barrels at 4° C until used. Prior to dredging, contami-
nated sediment was collected from BRH (41°9' N and 73°13' W) with a gravity
2
box corer (0.1 m ) to a depth of 1.21 m, thoroughly mixed, press sieved
through a 2-mm sieve, and refrigerated (4° C) until used. Details of sediment
collection and storage procedures may be found in Rogerson, Schimmel, and
Hoffman (1985). In all experiments, sediments were allowed to reach test tem-
perature and were mixed prior to use.
Organism collection and holding
35. Two separate experiments were completed using oxidized REF and BRH
sediments. Mussels were collected from the Narragansett Bay reference popu-
lation (71°24.0' W by 41°29.4' N) with a scallop dredge from a depth of 10 m.
Collection information for each experiment is listed in Table 1. The animals
Table 1
Collection Information for the Mussels Used
in the Laboratory Experiments
Collection Experiment Temperature Salinity
Experiment Date Begun "C g/kg
1 17 Jan 85 05 Feb 85 2.0 30.0
2 22 Feb 85 12 Mar 85 5.0 30.0
19
-------
were sorted to obtain a size range of 50- to 55-mm shell length and acclimated
in flowing unfiltered Narragansett Bay seawater at a rate of 1° C per day to
Suspended sediment dosing system
36. Laboratory studies required the construction of two identical
sediment dosing systems to provide either BRH or REF material as suspended
sediment simultaneously. Each dosing system (Figure 3) consisted of a
conical-shaped slurry reservoir placed in a chilled fiberglass chamber, a
diaphragm pump, a 4-fc separatory funnel, and several return loops that di-
rected the particulate slurry through dosing valves. The slurry reservoirs
(40 cm in diameter by 55 cm high) contained 40 £ of slurry composed of 37.7 I
of filtered seawater and 2.3 I of either BRH or REF sediment. The fiberglass
chamber (94 by 61 by 79 cm high) was maintained between 4° and 10° C using an
externally chilled water source to minimize microbial degradation during the
test. Polypropylene pipes (3.8-cm diam) extended to the bottom of the reser-
voir cones and were connected to pumps (16- to 40-Jt/min capacity) fitted with
Teflon diaphragms. These pumps were used to circulate the slurry while mini-
mizing abrasion which might produce changes in the physical properties (e.g.,
particle size) of the material.
15° C
TO EXPOSURE
SYSTEM
DOSING
VALVE
Figure 3. Suspended sediment
dosing system
20
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37. The slurry was pumped up to separatory funnels and returned via an
overflow to the reservoir through polypropylene pipes. The separatory funnel
provided the constant head pressure needed to circulate the slurry through
Teflon tubing to the dosing valves where the slurry was mixed with seawater to
provide the desired concentrations for the toxicity tests. Narragansett Bay
seawater filtered (to 15 u) through sand filters was used.
Suspended sediment oxidation system
38. The REF and BRH sediments used in these experiments were oxidized
prior to introduction into the dosing system. The objective of this portion
of the FVP was to evaluate the relationship between biological endpoints mea-
sured in the laboratory and the field. The field collections of sediment in-
dicated rapid oxidation of the surficial BRH sediments on the disposal mound.
Because the most likely source of particulate contaminants in the water column
was the oxidized surficial sediment, it was decided that laboratory exposures
would be conducted with BRH sediment that had been oxidized in a consistent
manner.
39. In order to obtain consistent states of oxidation for both REF and
BRH sediments, 2 I of sediment were transferred to an inverted polycarbonate
carboy and diluted to 19 £ with filtered natural seawater at room temperature
and aerated for 3 to 4 days (Figure 4). The contents were transferred to the
composite dosing system reservoir and diluted to 38 I with natural seawater.
Chemical oxygen demand measurements indicated that this time period was suffi-
cient to satisfy the immediate oxygen demand of the sediments.
Exposure system
40. An exposure system was constructed to provide a constant concen-
tration of suspended sediment to mussels in the laboratory. This system con-
sisted of recirculating loops from the suspended sediment dosing system which
were connected to a dosing valve at each exposure chamber. The concentration
of total suspended particulates was maintained at approximately 12 mg/ml in
both the REF and BRH loops. The exposure system was capable of delivering
either REF or BRH sediment directly into each mussel exposure chamber via a
dosing valve. The combined use of a REF and a BRH dosing valve at an exposure
chamber allowed delivery of a mixture of the two sediments. The percent con-
centrations of BRH and REF sediment varied between treatments; however, a
total suspended sediment concentration of approximately 10 mg/i (dry weight)
was maintained in all five laboratory exposure treatments. This concentration
21
-------
oir line
was chosen because it approximated the background field suspended sediment
concentration at the CLIS disposal site.
41. Each mussel exposure chamber was equipped with a transmissometer,
an instrument capable of measuring light attenuation due to suspended sediment
in the chamber (Figure 5). The dosing valves for each treatment were con-
trolled by a transmissometer-microprocessor feedback loop (Sinnett and Davis
1983). The transmissometer in each chamber was calibrated by regressing
suspended sediment concentrations, measured by filtration onto glass fiber
filters, with the transmissometer units displayed on a microprocessor. A
transmissometer value was calculated that corresponded with the desired sus-
pended sediment concentration of 10 mg/i for each chamber. As the mussels re-
moved suspended sediments, the microprocessor opened dosing valves to deliver
additional suspended sediment at 2-min intervals. In this manner, suspended
sediment concentrations were maintained at the desired values (±10 percent).
The transmissometer circuit was also connected to a strip chart recorder which
allowed the operation of the system to be monitored continuously. Each cham-
ber was aerated with three 25- by 2.5-cm air stones to provide sufficient
oxygen and to ensure even distribution of suspended particulates (Figure 5).
22
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SEAWATER/SEDIMENT
SLURRY
ALGAE
J5=
TO MICROPROCESSOR
TRANSMISSOMETER
Figure 5. Laboratory exposure
system
MIXING
CHAMBER
I
RECIRCULATING (f^
PUMP
42. In addition to the suspended sediment, food in the form of a uni-
cellular alga, Isochrysis galbana, was supplied to each exposure chamber.
Periodic measurements were made of mussel clearance rates in each chamber to
determine the volume of algae required to maintain an algal concentration of
0.5 mg/Jl. This concentration constituted an adequate maintenance ration for
the mussels. Algae were added at 5-min intervals by means of a peristaltic
pump. All experiments were conducted at 15° C with filtered seawater that
flowed through each experimental chamber at a rate of 0.4 Jl/min. Each chamber
was cleaned every other day.
43. The purpose of the laboratory experiments was to expose M. edulis
to a range of BRH concentrations that may have been present in CLIS and to
assess the biological effect on these organisms. Mytilus edulis were exposed
for approximately 1-month periods at the CLIS disposal site; therefore, expo-
sures of similar duration, 28 days, were used for the laboratory exposures.
44. At the start of both experiments, 150 mussels were placed into each
chamber. Mytilus edulis were sampled at time zero to determine initial tissue
residue concentrations, and for SFG testing.
45. Experiment 1 consisted of three exposure concentrations: 100-,
50-, and 0-percent BRH suspended sediment. Mytilus edulis were removed from
each treatment on Day 14 for chemical and biological analysis. Experiment 1
23
-------
was terminated at Day 14 because adverse biological effects (e.g., reduced
filtration rate) were observed in both treatments containing BRH sediment.
46. Experiment 2 was conducted with lower concentrations of BRH sus-
pended sediment. Exposure concentrations of suspended sediment in Experi-
ment 2 were 30-, 10-, and 0-percent BRH. Fifteen organisms were removed on
Days 7, 14, 21, and 28 for tissue residue analysis. Whole water chemistry
samples were taken within 1 day of organism sampling. Dissolved and particu-
late water samples were taken within 24 hr of Days 0, 14, and 28. Mussels
were sampled on Days 14 and 28 for biological analysis. In addition, a water
sample was taken on Day 29 to evaluate the performance of the system without
any mussels in the exposure chambers.
47. The operation of the system (dosing valves, flow rates, etc.) was
monitored daily. Experiments using the 100- and 0-percent BRH concentrations
required only one dosing valve each, while the 50-percent BRH treatment re-
quired a REF and BRH valve that delivered equal amounts of suspended material.
A strip chart record for each treatment indicated that the dosing valves were
operating properly. The 10- and 30-percent BRH treatments also required two
dosing valves per treatment; however, the REF and BRH dosing valves delivered
different amounts of suspended material. This was accomplished by adjusting
the delivery volume of each valve. The mixture of BRH and REF material was
checked daily and adjusted if necessary.
SFG procedures
48. Calculation of the SFG index for M. edulis required the measurement
of four parameters: clearance rate, respiration rate, food absorption effi-
ciency, and ammonia excretion rate. Measurements were completed under stan-
dardized conditions: temperature 15° C, salinity 30 ppt, and an algal
concentration of 0.5 mg/&. All SFG measurements for a given treatment were
completed in the order shown below within 28 hr after termination of the ex-
periment. The detailed methods are described by Nelson, Black, and Phelps
(1985); therefore, only a brief summary of each procedure is provided here.
49. Clearance rate. Clearance rate is defined as the volume of water
completely cleared of particles >3 p in some unit of time (Widdows, Fieth, and
Worrall 1979). Mussels were placed into individual chambers through which 1 y
filtered seawater flowed at a rate of 75 ml/min. The unicellular alga T-Iso
was added to the filtered seawater to deliver an incoming cell concentration
of approximately 25,000 cells/ml (about 0.5 mg/£) to each chamber. Each
24
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chamber was gently aerated to ensure that complete mixing and no settling of
algae occurred. Mussels were allowed to acclimate in the chambers for at
least 1 hr prior to any measurements. Incoming and outgoing particle concen-
trations for each chamber were then measured 3 times at 1-hr intervals with a
Coulter Counter (Model TAII).
50. Respiration rate. Respiration rates were determined by isolating
each mussel in a glass respirometer vessel fitted with a P02 electrode. The
electrode was connected to a Radiometer oxygen meter (Model PHM71) which was
in turn connected to a strip chart recorder. The decline in P02 was monitored
on a strip chart recorder for approximately 30 min. Seawater containing algae
(0.5 mg/jl) was pumped into the vessel during an acclimation period at a rate
of 80 ml/min to ensure that food was present in the chamber and that routine
metabolic rate was measured.
51. Absorption efficiency. After completion of the respiration rate
measurements, all fecal material was removed from each feeding chamber. This
ensured that only the algae consumed during the SFG procedures were used in
the absorption efficiency measurements. At the food concentration used in the
SFG measurements, approximately 0.5 mg/l, no pseudofecal production occurred.
The mussels were allowed to feed overnight in the chambers, and the feces were
collected from each chamber the following morning. Fecal material was dried
for 24 hr at 100° C, weighed, ashed at 500° C for 4 hr, and reweighed to
determine the ash-free dry weight:dry weight ratio. A similar procedure was
completed with the cultured algae to obtain the ash-free dry weight:dry weight
ratio of the food. Absorption efficiencies were calculated for each treatment
according to the method of Conover (1966).
52. Ammonia excretion rate. Mussels were placed individually into
HCl-stripped beakers containing 300 ml of 1-y filtered seawater for a period
of 3 hr. Mussels were then removed and a 0.45-y filtered, 50-ml sample was
collected from each beaker, deposited into acid-stripped polyethylene bottles,
and stored in a freezer at -20° C until analyzed. Ammonia analyses were com-
pleted in duplicate for each sample according to the method of Bower and Holm-
Hansen (1980).
SFG calculations
53. After completion of the physiological measurements, the length and
volume of each mussel were measured and the tissue excised, dried for 24 hr at
100° C, and weighed. The clearance rates, respiration rates, and ammonia
25
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excretion rates were standardized to the mean weight of all the mussels used
in the treatment. This procedure was used instead of standardizing to a 1-g
mussel by using allometric equations because all mussels were approximately
the same length and weight. The use of allometric equations is necessary only
when mussels of variable size and weight are used (Bayne, Clark, and Moore
1981) .
54. The weight-standardized values for each mussel were then used to
calculate the SFG of each individual by substitution into the following
equation:
SFG = (C x A) - (R + E) (5)
where
C = energy consumed (clearance rate x surrounding food concentra-
tion x energy of food)
A = absorption efficiency
R and E = energy lost through respiration and nitrogen excretion,
respectively
The following energy conversions were used to calculate SFG:
1 mg of T-Iso =4.5 x 10^ cells (this experiment)
1 mg of T-Iso = 19.24 J (this experiment)
1 ml (>2 respired = 20.08 J (Crisp 1971)
1 mg NH4~N = 24.56 J (Elliot and Davidson 1975)
The energy content of T-Iso was determined by filtering a volume of the algae
onto preweighed glass fiber filters, drying them at 100° C for 24 hr, and
reweighing them to determine algal dry weight. They were analyzed then using
the dichromate wet oxidation method of Maciolek (1962) to determine oxygen
consumed and the resultant energy content.
Clearance rates in
laboratory exposure system
55. In the laboratory documentation portion of the FVP (Nelson, Black,
and Phelps 1985) it was noted that the main factor contributing to lowered SFG
values in BRH-exposed mussels was decreased clearance rates. Therefore, it
was decided to monitor the clearance rates of mussels in the exposure chambers
during this set of experiments in order to observe when an adverse biological
effect first began. The clearance rates were estimated using transmissometer
readings (TRNUM) in each exposure chamber. As previously described, the TRNUM
26
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was calibrated to the level of suspended particulates in each chamber. An
initial transmissometer reading was taken for each exposure chamber. The
dosing valve was then shut off for a period of 10 min, after which the final
TRNUM was recorded. In this manner the decrease in suspended particulate
level (milligrams/litre) was determined as the difference between the initial
and final concentrations. This value was multiplied by the chamber volume
(100 I) to determine the total milligrams of material removed from the cham-
ber. In addition to suspended particulates being removed by the mussels, some
material was lost down the overflow. This was accounted for by multiplying
the flow rate (0.4 8,/hr) by the mean particulate level during the 10-min pe-
riod. This value was subtracted from the total volume removed to determine
the volume of material removed by the mussels (MGS). Because of the design of
the chamber, very little settling out of suspended sediment occurred, so this
possible route of sediment loss during the 10-min time period was not factored
in. The clearance rate (CR) was then calculated using the following formula:
The clearance rates were measured in this manner for each treatment on Day 9
in the first experiment and Days 7 and 16 in the second experiment. The CR
measurement on Day 16 was taken twice. The first one was completed immedi-
ately after the exposure tanks were cleaned when it was observed that the
algae pump was not on. The pump was then turned on, the system allowed to
equilibrate for 1 hr, and the CR measurement repeated. Both of these measure-
ments were included to show the possible loss of an initial "active feeding"
response in the mussels from the 30-percent BRH treatment when food was first
provided.
Actual growth
56. The actual growth of 10 mussels was measured from each treatment
for comparison with SFG values. This was accomplished by numbering 15 mussels
(extras included in case of mortality) in each treatment prior to the start of
the experiment, recording the length, and remeasuring the mussels on Day 14 in
the first experiment and Days 14 and 28 in the second experiment. In this
manner, growth data were obtained separately for the first and second 14-day
periods in the second experiment.
CR (litre/hour/mussel) = MGS *
60 min/hr .
10 min
Mean Total number
t mg/i, t mussels
per chamber
(6)
27
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Field Methods
Organism collection and holding
57. All mussels used in the field studies for the FVP were collected by
scallop dredge from Narragansett Bay. In general, M. edulis were collected
1 to 2 days prior to field deployment to Long Island Sound. They were re-
turned to the laboratory where 100 5- to 7-cm organisms were sorted and placed
into polyethylene baskets. Each basket was placed in holding tanks of flowing
unfiltered seawater until deployed in the field.
Exposure
58. Mytilus edulis were deployed at CNTR, 400E, 1000E, and REFS at the
CLIS disposal site (Figure 2). The physical arrangement of each station is
detailed by Phelps and Galloway (1980). In short, each station consisted of a
surface buoy attached by cable to a concrete mooring on the bottom, with two
smaller satellite moorings attached to the larger main mooring. A subsurface
buoy was attached to each small mooring from which the mussel baskets were
hung 1 m above the bottom. Two baskets were attached to each subsurface buoy
at each deployment.
59. The deployments of M. edulis at the CLIS disposal site are summa-
rized in Table 2. Mussels were deployed at each station for a period of
1 month predisposal to collect baseline data (Cruise number T - 04). A second
deployment occurred during disposal operations, except that no mussels were
placed at the CNTR station (T = 0, T + 2). Mussels were deployed for 1-month
periods over the next 3 months (T + 8, T + 12, T + 15), then on a quarterly
basis for the next year (T + 27, T + 43, T + 74, and T + 116). In addition,
several sets of mussels were left at each station for 7 months (T + 21). The
cruise number designation is not related to the length of deployment.
60. Mytilus edulis were retrieved from the subsurface buoys by divers.
Mussels used for chemical analysis were frozen immediately. The remaining
mussels were maintained in tanks of flowing seawater on deck and returned to
ERLN later that day and held in flowing unfiltered seawater overnight. The
next morning, mussels were distributed to the appropriate investigators for
biological analyses.
61. Estimated field exposure via tissue residues. Exposure conditions
present in the field during each mussel deployment were not as well charac-
terized as they were in the laboratory studies. As a result, the description
28
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Table 2
Cruise Number, Deployment Date, Retrieval Date, and Length of
Deployment for Mussels Transplanted to CLIS
Cruise Number
Weeks*
Deployment
Date
Retrieval
Date
Length of
Deployment
T -
04
16
Mar
83
22
Apr
83
1
month
T =
0**
22
Apr
83
24
May
83
1
month
T +
2
23
Apr
83
07
Jun
83
6
weeks
T +
8
07
Jun
83
13
Jul
83
1
month
T +
12
13
Jul
83
10
Aug
83
1
month
T +
15
10
Aug
83
06
Sep
83
1
month
T +
21
16
Mar
83
18
Oct
83
7
months
T +
27
06
Sep
83
29
Nov
83
3
months
T +
43
29
Nov
83
20
Mar
84
4
months
T +
55
18
Oct
83
05
Jun
84
8
months
T +
74
12
Jun
84
17
Oct
84
4
months
T +
116
11
Jul
85
14
Aug
85
1
month
* The "cruise number, weeks" is not related to the "length of deployment."
** T = 0 refers to the termination of disposal activities at the FVP site on
18 May 1983.
of M. edulis exposure to BRH material in the field is more qualitative than
quantitative and will be presented in two parts. First, a prediction of field
exposure is based on mussel tissue residues. The relationship between expo-
sure to BRH sediments and tissue residues was determined in the laboratory
experiments. Tissue residues from the 0-, 10-, and 30-percent BRH treatments
at 28 days were regressed against measured BRH exposure concentrations (0,
1.5, and 3.3 mg/£) from the same exposures. In order to correct for back-
ground residues in the laboratory, the PCB concentration of the 0-percent BRH
treatment was subtracted from the others prior to regression analysis. The
resultant equation, milligrams per litre of BRH material = (PCB residue
2
x 0.000965) - 0.0019 , R ¦ 0.999 , was then used to calculate the average
sustained concentration of BRH material necessary to achieve the residue value
obtained in the field. The estimated BRH exposures in the field were deter-
mined by substituting the mussel PCB tissue residue concentration directly
29
-------
into the above equation. This estimate was assumed to represent an upper
range of suspended BRH material present. A second estimate was determined by
first subtracting the PCB concentration in mussels at the REFS station from
the other stations during that collection. This removed the Long Island Sound
background PCB levels from the estimates, and thus was assumed to represent a
lower range of BRH present in CLIS. This procedure was completed for each
collection date and station at which mussels were retrieved.
62. Estimated exposure via water chemistry data. A second estimate of
exposure was generated from the PCB and Cu concentrations in the whole water
samples collected during various postdisposal cruises. The concentration of
BRH material that would have to be present to produce these levels was de-
termined by dividing the concentration of PCB and copper present in the barrel
material collected from BRH (2,900 Mg/g and 6,910 ng/g for Cu and PCB, respec-
tively) . A range of exposures was also calculated for the water chemistry
data; estimated BRH material was determined with and without subtracting the
concentration at the REFS station.
SFG procedures
63. SFG was measured the morning after mussels were returned from the
field. The SFG procedures used to analyze the field mussels were the same as
those used to measure the laboratory mussels. Physiological measurements of
the field-collected mussels were conducted at the same ambient temperature
they were exposed to in the field. Ambient water temperatures in Narragansett
Bay were within 1° to 2° C of those at the CLIS disposal site.
Statistical analysis
64. The primary objective of the FVP was to compare laboratory and
field responses under similar conditions. The highly dynamic temporal and
spatial conditions in the field made it Impossible to replicate these condi-
tions in the laboratory. Consequently, the experimental design employed was
such that qualitative relationships were made between the laboratory and
field. Therefore, descriptive statistical procedures were used, not inferen-
tial tests. Standard errors presented In tables and figures were calculated
from 10 samples within a treatment or basket. These values are included to
indicate variability within a treatment, not among 10 statistical replicates.
65. Statistical analysis of the data was completed in three parts,
exposure-effects, residue-effects, and laboratory-to-field comparison. In the
laboratory experiments, regression analysis was used to determine the
30
-------
relationship between SFG and BRH exposure concentration (Snedecor and Cochran
1967). The limited exposure data from CLIS precluded measuring similar rela-
tionships for the field mussels. The relationship between SFG and tissue
residue in the laboratory experiments was determined by regressing the mean
SFG value for each treatment against the corresponding mean tissue residue
(Snedecor and Cochran 1967). Data from both laboratory experiments were
included. This procedure was completed individually for each of the 10 se-
lected chemical contaminants and the two summary statistics.
66. Prior to making comparisons between laboratory and field effects,
it was necessary to establish whether exposures (i.e., residues) were similar
in the laboratory and the field. This was accomplished by examining the tis-
sue residues of all mussels from laboratory and field exposures together,
independent of exposure concentration or station location and date. The PCB,
ethylan, PAH, and SUM and CENT variables were analyzed by cluster analysis
(BioMedical Programs 1983) to establish which tissue residues, among all the
laboratory treatments and field stations, were most similar. Prior to analy-
sis, residue values for each compound were normalized using standard devia-
tions from the mean. This procedure ensured that each variable was weighted
equally, even if its absolute value was orders of magnitude different from
another variable.
31
-------
PART III: RESULTS
Laboratory
Exposure
67. System monitoring. The M. edulis exposure system was monitored for
both total suspended solid (TSS) concentrations and the percentage of REF and
BRH sediments. The strip chart record indicated that the system maintained a
suspended particulate concentration of 10 mg/£ approximately 90 percent of the
time. Examples of times when the 10 mg/£ was not maintained include periods
when exposure tanks were cleaned, slurry reservoirs were changed, and lines
were clogged. Overall, the system provided a nearly constant total suspended
particulate concentration to the mussels. The concentration of BRH sediments
dosed into each treatment is listed in Table 3.
Table 3
Suspended Sediment Concentrations in the Mussel Exposure System*
Nominal
Measured
Calculated
Percent
Percent
BRH Sediment
BRH
BRH**
mg/£
100
100 (0.00)
10.0
50
50 (0.83)
5.0
30
33 (0.84)
3.3
10
15 (1.39)
1.5
0
0 (0.00)
0.0
* Values include nominal and actual BRH suspended sediment concentration.
** Standard error in parentheses.
68. When the TSS concentration dropped in the 50-percent BRH exposure
tank, a pulse of equal length was sent to both the REF and BRH dosing valves.
Volumetric measurements of the BRH and REF sediment doses indicated that equal
amounts (±5 percent) of BRH and REF material were delivered to the 50-percent
BRH exposure chamber.
69. The 10- and 30-percent BRH treatments required two dosing valves
per treatment. Because the pulse length could not be adjusted separately for
32
-------
each valve, manual adjustment of each valve was required to provide the de-
sired percent concentration. The volumetric amount of BRH and REF material
delivered to each treatment was monitored and recorded (Table 3). In the
treatment with a nominal concentration of 10-percent BRH, the actual value
delivered was 15 percent. In the 30-percent BRH treatment, the actual value
was 33 percent BRH.
70. Chemical monitoring. The results of the chemical monitoring are
prefaced by a brief restatement of the purpose of the exposure system to aid
in the understanding of the results. The system used in this experiment was
designed to maintain a constant particulate concentration of 10 mg/£ in the
exposure chambers. Initially, 150 animals were placed into each chamber with
clearance rates of approximately 2 A/mussel/hr, or a total of 300 &/hr. The
seawater flow rate through each chamber, independent of suspended sediment
additions, was approximately 24 &/hr. In effect, suspended sediment was added
at a rate 12.5 times that of seawater to each exposure chamber each hour to
compensate for sediment removed by the mussels. This has important conse-
quences on the behavior of the contaminants in the exposure system.
71. If all the contaminants were associated with the suspended sedi-
ment, the contaminant concentrations in the exposure chambers should be simi-
lar to those predicted by regressing the TSS concentrations with contaminant
concentrations in the BRH material. Conversely, any contaminants that do not
remain bound to the particulates could attain concentrations in the exposure
system different from those predicted from the TSS data. This occurs because
the mussels in the system are more efficient at removing the particulate-bound
contaminants than they are at removing the dissolved contaminants. This the-
ory is proposed to explain the results of the chemical concentrations in the
exposure system, using PCB and copper as examples.
72. Whole water samples were taken for chemical analysis on Days 1, 7,
14, 21, and 28 in the second experiment. The mean PCB concentrations
(nanogram/litre) for the five sampling dates for each exposure treatment in
the second experiment are given in Table 4. The corresponding concentration
of BRH sediment was estimated by regressing the nominal concentration of BRH
against the expected value of PCB. Expected PCB concentrations were based on
the PCB concentrations in the BRH sediment (6 ng/mg) plus background seawater
concentrations. Substitution of the actual measured values of PCBs in the
exposure system into the equation provided an estimated value of the
33
-------
Table 4
Chemical Monitoring of the Exposure System In Experiment 2
Nominal Treatment PCB Concentration, ng/7 BRH Concentration, mg/T
Concentration, % BRH Expected Measured Estimated Measured
0 1.1 2.2 0.2 0
10 7.1 11.9 1.8 1.5
30 18.8 23.6 3.8 3.3
concentration of BRH sediment in the system. The estimated concentration of
BRH sediment in each treatment is similar to the actual measured values.
These data suggest that PCB concentrations in the system are closely associ-
ated with the TSS concentrations.
73. Copper concentrations were measured both with and without mussels
in the exposure system at 10 mg/jl TSS for each treatment. With no mussels in
the exposure system, the total copper concentrations were 9.37 and 2.5 yg/£
for the 30- and 10-percent BRH treatments, respectively. These concentrations
represent 3.8 and 1.8 mg/£ BRH sediment in the two treatments, respectively.
Under these conditions, the predicted and measured copper concentrations were
comparable. This resulted because the effective flow of suspended sediment
and incoming seawater is the same. The only loss of TSS was out the overflow
due to seawater flow rates.
74. When mussels were present in the system, the mean copper concentra-
tions were 17.0 and 10.7 yg/Jl for the 30- and 10-percent BRH treatments, re-
spectively. These copper concentrations correspond to 68- and 43-percent BRH
sediment in the two treatments, respectively, and conflict with those expected
from the TSS data. The results may be explained by the fact that copper, due
to its solubility in seawater, became disassociated from the TSS because sus-
pended solids were delivered at a higher rate to the exposure chamber than the
rate of incoming seawater. When a dose of BRH suspended sediment was deliv-
ered to an exposure chamber, all contaminants were introduced at the same
rate. Because the mussels were more efficient at removing particulates than
dissolved contaminants, dissolved copper tended to accumulate, which resulted
in higher concentrations of copper than those predicted from the TSS data
alone.
34
-------
Tissue residue
75. Differences in contaminant concentrations between BRH and REF sedi-
ments facilitated the tracking of these contaminants in exposed biota (Appen-
dix B). Results of Experiment 1 indicate that PCB tissue concentration in
mussels is directly related to exposure concentration (Table 5). PCBs in mus-
sels from the 0-percent BRH concentration remained about the same over the
14-day experiment.
Table 5
PCB Tissue Residues (ng/g dry weight) in Mussels from
Laboratory Experiment 1
PCB Residue in Indicated Percent BRH Sediment
Day 0 50 100
0 117 117 117
14 154 2,100 3,700
76. The PCB residue data from Experiment 2 are listed in Table 6 and
graphically depicted in Figure 6. Tissue residues, measured at 7-day inter-
vals, indicated that the mussels in the 0-percent BRH chamber maintained a
relatively constant background concentration of PCBs throughout the experi-
ment. In the 10- and 30-percent BRH chambers, the concentration of PCBs in
the mussels increased from Day 0 to Day 14, then remained nearly constant
Table 6
PCB Tissue Residues (ng/g dry weight) in Mussels from
Laboratory Experiment 2
PCB Residue in Indicated Percent BRH Sediment
Day 0 10 30
0 210 210 210
7 280 1,110 2,100
14 270 1,910 3,600
21 360 1,720 3,600
28 280 1,840 3,700
35
-------
SAMPLING DAY
Figure 6. Concentrations of PCB as A1254 in
the tissue of M. edulis exposed to BRH
suspended sediment for 28 days
between Days 14 and 28, suggesting that the mussels reached a steady state
somewhere between Days 7 and 14. The steady-state PCB concentration in the
30-percent BRH mussels was almost double that of the mussels from the
10-percent BRH treatment. The actual concentration of BRH dosed to the
30-percent BRH treatment, 3.3 mg/JI, is nearly double that dosed to the
10-percent BRH mussels, 1.5 mg/£. The measured whole water PCB concentrations
were 11.86 and 23.57 ng/fc for the 10- and 30-percent BRH treatments, respec-
tively. These data indicate a good relationship between the actual dosed
concentrations of BRH suspended sediment, the measured whole water concentra-
tions, and the PCB tissue residues in the mussels in Experiment 2.
77. A comparison of the tissue residues between the two experiments can
be made for Days 0 and 14. The PCB concentration in the Day 0 mussels from
Experiment 1 was almost half that in Day 0 mussels from Experiment 2 (117 and
210 ng/g, respectively). In addition, Day 14 PCB concentrations were about
the same for the 10- and 50-percent BRH exposed mussels (1,910 and 2,100 ng/g)
as well as the 30- and 100-percent BRH exposed mussels (3,600 and 3,700 ng/g).
These data show dose responses within each experiment; however, there is poor
agreement between experiments. The PCB data from these experiments were nor-
malized to nanograms/gram of lipid, and the results are presented in Table 7
36
-------
Table 7
PCB Concentrations (ng/g Lipid) in Mussels from
Both Laboratory Experiments
PCB Concentration in Indicated Percent BRH Sedir.ent
Day 0* 0** 10** 30** 50* 100*
0 2,900 2,400 2,400 2,400 2,900 2,900
7 —t 5,200 17,100 24,000
14 3,800 4,300 27,000 54,000 53,000 119,000
21 — 5,000 35,000 67,000
28 — 3,800 30,000 66,000
* Experiment 1.
** Experiment 2.
t Not sampled.
and Figure 7. Inspection of these data shows that differences between experi-
ments can be explained when differences in lipid content of the organisms are
taken into account. In addition, this procedure indicates that *a dose-
response relationship does exist between experiments when the Day 14 data from
both experiments are combined (Figure 7).
EXPOSURE CONCENTRATION (X BRH)
Figure 7. Concentrations of PCB as A1254, nor-
malized for lipids, in the tissue of M. edulis
exposed to BRH sediment for 14 days
37
-------
78. In addition to PCBs, tissue residues of phenanthrene, the sum of
the 178 alkyl homologs, fluoranthene, benzo(a)pyrene, ethylan, cadmium, cop-
per, chromium, and iron were also measured on Days 0, 7, 14, 21, and 28 of
Experiment 2. The summary statistics, SUM and CENT, of the PAHs were also
calculated for each of these sampling dates. These data are summarized graph-
ically in Figures 8-13.
79. While each of the graphs presented as Figures 8-13 will not be
discussed at length, it is interesting to note the relationship between the
molecular weight of the organic compounds and tissue residue over time. The
benzo(a)pyrene tissue residues follow a pattern similar to that of PCB. After
7 days, residues remain nearly constant for each exposure concentration. The
fluoranthene residues are initially higher in the 30-percent BRH treatment.
However, they decrease over the 28-day exposure period to a level comparable
to the 10-percent BRH treatment. Residues for both of these treatments are
elevated compared with the 0-percent BRH mussels. Phenanthrene, an even lower
molecular weight PAH, increased initially but then decreased in both the
30- and 10-percent BRH treatments to a level comparable to the 0-percent BRH
exposure. These data would suggest that mussels have the ability to
metabolize and/or excrete the lower molecular weight PAHs, even during
continuous exposure. In addition, the data would indicate that only higher
molecular weight compounds should be used to relate exposure levels and subse-
quent tissue residue levels because, even under relatively constant exposure
conditions, residues of lower molecular weight PAHs did not reflect exposure
concentrations.
Biological effects
80. Clearance rates in the exposure system. Clearance rate measure-
ments in the exposure system are listed in Table 8. In the first experiment,
clearance rates were dramatically reduced in both the 50- and 100-percent BRH
treatments after 9 days of exposure. This observation was reinforced by the
strip chart record which indicated a reduced number of doses provided to these
two treatments.
81. Similar results were observed in Experiment 2. Measurements on
Day 7 indicated reduced clearance rates in both the 30- and 10-percent BRH
treatments when compared with the 0-percent BRH treatment. On Day 16 this
measurement was repeated soon after the exposure system was cleaned. The par-
ticulate levels were at the proper concentration; however, it was later noted
38
-------
a. Phenanthrene
SAMPLING DAY
b. 178 alkyl homologs
Figure 8. Concentrations of phenanthrene and
178 alkyl homologs in the tissue of M. edulis
exposed to BRH suspended sediment for 28 days
39
-------
SAMPLING DAY
a. Fluoranthene
SAMPLING DAY
b. Benzo(a)pyrene
Figure 9. Concentrations of fluoranthene and
benzo(a)pyrene in the tissue of M. edutis ex-
posed to BRH suspended sediment for 28 days
40
-------
SAMPLING DAY
a. SUM of PAHs
SAMPLING DAY
b. CENT of PAHs
Figure 10. Concentrations of the SUM of PAHs and
CENT OF PAHs in the tissue of M. edulis exposed
to BRH suspended sediment for 28 days
41
-------
SAMPLING DAY
a. Ethylan
SAMPLING DAY
b. PCB
Figure 11. Concentrations of ethylan and PCB
as A1254 in the tissue of M. edulis exposed
to BRH suspended sediment for 28 days
42
-------
SAMPLING DAY
a. Cadmium
SAMPLING DAY
b. Copper
Figure 12. Concentrations of cadmium and
copper in the tissue of M. edulis exposed
to BRH suspended sediment for 28 days
43
-------
7-
• TIME ZERO
O 0% BRH
~ !0% BRH
A 30% BRH
I-
"I —I
7 14
SAMPLING DAY
a. Chromium
i
21
28
750
600-
450
300
150-1
• TIME ZERO
0 0% BRH
~ 10% BRH
A 30% BRH
SAMPLING DAY
b. Iron
Figure 13. Concentrations of chromium and
iron in the tissue of M. eduZis exposed to
BRH suspended sediment for 28 days
44
-------
Table 8
Clearance Rates (litres/hr/mussel) of Mussels
in Dosing System Exposure Tanks
Day
0% BRH
50% BRH
100% BRH
Experiment 1
9
2.22
o
o
0.10
0% BRH
10% BRH
30% BRH
Experiment 2
7
1.60
0.52
0.26
16
(no algae)
1.23
0.90
0.82
16
(algae on)
2.87
2.55
0.94
that the pump supplying algae to the exposure tanks was off. At this time,
the clearance rates in the 10- and 30-percent BRH treatments were again lower
than the 0-percent BRH treatment. The algae pump was then turned on, and
algae levels were allowed to equilibrate (about 1 hr). Clearance rate
measurements were repeated, and this time the 0- and 10-percent BRH mussels
both increased their clearance rates considerably to 2.87 and 2.55 A/hr,
respectively, while the clearance rate of the mussels in the 30-percent BRH
treatment remained about the same as when no algae were present. Mussels are
known to increase their feeding rate in the presence of food after being
starved. Mussels from the 10-percent BRH treatment were able to increase
their feeding rate to a level comparable to mussels from the 0-percent BRH
treatment after the algae were added, while those in the 30-percent BRH
chamber were not. These data indicate that mussels from the 30-percent BRH
treatment were more severely impacted than those mussels from the 10-percent
treatment.
82. SFG measurements. The values for the physiological parameters have
been standardized to the mean weight of all the mussels for a particular ex-
periment. The mean weight for the mussels from Experiment 1 was 0.48 g and
for Experiment 2 was 0.74 g.
83. Clearance rates. The clearance rate data are listed in Table 9.
The mussels from the 50- and 100-percent BRH chambers exhibited lower
45
-------
Table 9
Clearance Rates of Mussels from the Two Laboratory Exposures*
Treatment
% BRH
Clearance Rate
£/hr
Experiment 1, Day 14
0
50
100
4.69 (0.25)
0.54 (0.20)
0.17 (0.07)
Experiment 2, Day 14
0
10
30
4.47 (0.18)
2.48 (0.56)
0.81 (0.30)
Experiment 2, Day 28
0
10
30
3.51 (0.43)
1.80 (0.41)
1.07 (0.24)
* Each value represents the mean of 10 mussels (standard error in
parentheses).
clearance rates after 14 days than the 0-percent BRH mussels in Experiment 1.
In Experiment 2, on Day 14, mussels from the 30-percent chamber exhibited re-
duced clearance rates compared with the 10-percent BRH mussels, which were, in
turn, lower than the 0-percent mussels. By Day 28, however, the clearance
rates of mussels from the 30- and 10-percent BRH chambers were not much dif-
ferent from each other but were still lower than the 0-percent BRH mussels.
84. Absorption efficiency. Mussel absorption efficiencies are listed
in Table 10. Only one pooled value was determined for each treatment. The
results indicate that there were no large differences in absorption efficiency
among the various exposure chambers.
85. Respiration rates. The respiration rate data are listed in
Table 11. Differences among chambers were relatively small at all of the sam-
pling times.
86. Ammonia excretion rates. The ammonia excretion rate values are
listed in Table 12. The largest variations among the chambers occurred in
Experiment 1, where the range of BRH exposure was the greatest.
46
-------
Table 10
Absorption Efficiencies of Mussels from the Two Laboratory Exposures
Treatment
Absorption
% BRH
Efficiency, %
Experiment
1, Day
14
0
78
50
80
100
85
Experiment
2, Day
14
0
88
10
84
30
87
Experiment
2, Day
28
0
72
10
78
30
85
Table 11
Respiration Rates of Mussels from the Two Laboratory Exposures*
Treatment
% BRH
Respiration Rate
ml 02/hr
Experiment
1, Day 14
0
50
100
Experiment
2, Day 14
0.41 (0.07)
0.39 (0.03)
0.41 (0.06)
0
10
30
Experiment
2, Day 28
0.36 (0.02)
0.41 (0.03)
0.42 (0.02)
0
10
30
0.38 (0.02)
0.48 (0.06)
0.46 (0.03)
* Each value represents
parentheses).
the mean of 10
mussels (standard
error in
47
-------
Table 12
Ammonia Excretion Rates of Mussels from the Two Laboratory Exposures*
Treatment
Z BRH
Excretion Rate
Ug NH^-N/hr
Experiment
1, Day
14
0
50
100
Experiment
2, Day
14
10.31
17.45
20.56
(0.90)
(3.09)
(10.48)
0
10
30
Experiment
2, Day
28
12.69
11.48
11.33
(0.82)
(1.60)
(1.61)
0
10
30
9.52
15.06
11.90
(1.37)
(2.34)
(1.39)
* Each value represents the mean of 10 mussels (standard error in
parentheses) .
87. SFG index. The Day 14 SFG values in the first experiment were re-
duced in the 50- and 100-percent BRH chambers (Table 13). The SFG values in
the second experiment followed the same pattern as the clearance rates. By
Day 14, mussels from the 10-percent BRH chambers exhibited lower SFG values
than the 0-percent BRH mussels; however, their SFG values were higher than
those of the mussels from the 30-percent BRH chamber.
88. The graph of the Day 14 SFG values and exposure concentration (Fig-
ure 14) suggested that the relationship between these two variables was not
linear. Therefore, the SFG data were log 10 transformed prior to regression
analysis. To avoid negative values (i.e., -7.14 for the 100-percent BRH
treatment), each SFG number was increased by 8 prior to log 10 transformation.
Regression analysis of the data indicated a significant inverse relationship
2
(P < 0.001, R = 0.99) between log SFG and BRH exposure concentration.
89. On Day 28, the SFG of mussels from the 10- and 30-percent BRH
chambers were lower compared with those of the 0-percent mussels. However,
they were not different from each other. Because there were only three data
48
-------
Table 13
SFG Values of Mussels from the Two Laboratory Exposures*
Treatment
% BRH
SFG
(J/hr)
Experiment
1, Day
14
0
50
100
Experiment
2, Day
14
10.62
-4.26
-7.14
(1.10)
(1.54)
(1.30)
0
10
30
Experiment
2, Day
28
14.17
5.03
-2.82
(0.59)
(2.00)
(1.81)
0
10
30
7.16
0.14
-1.79
(1.86)
(1.30)
(1.39)
* Each value represents the mean of 10 mussels (standard error in
parentheses).
points, regression analysis was not appropriate. Visual inspection of the
data suggested a relationship between SFG and BRH concentration similar to
that observed on Day 14 (Figure 14).
90. Actual growth. In addition to the SFG determination, changes in
shell length were measured on mussels from the same chamber (Table 14). In
Experiment 1 the initial mean length of the mussels used was 5.02 ± 0.01 cm.
The initial mean mussel length in Experiment 2 was 5.00 ± 0.01 cm. The growth
increment is listed for the first 14-day period, in both experiments, and for
the second 14-day period in the second experiment. In Experiment 1, shell
growth of mussels from the 0-percent BRH chamber was greater than that of the
mussels from the 50- and 100-percent BRH chambers. In Experiment 2, mussels
in the 0-percent BRH chamber again showed a greater increase in shell growth
than mussels from the 10- and 30-percent BRH treatments. Actual shell growth
followed the same pattern as the SFG values and clearance rate measurements at
Day 14 in both Experiments 1 and 2 and at Day 28 in Experiment 2
(Figures 15-17).
49
-------
EXPOSURE CONCENTRATION (% BRH)
Figure 14. Relationship between the SFG of
M. edutis and BRH exposure concentration on
Days 14 and 28 of the laboratory experi-
ments. Data from Experiments 1 and 2 are
presented for Day 14
Table 14
Actual Growth of Mussels in the Exposure Chambers*
Treatment Growth, mm ~
% BRH "ays 0-14 Da7s l5'28
Experiment 1
0 0.40 (0.13)
50 0.11 (0.06)
100 0.06 (0.04)
Experiment 2
0 0.75 (0.14) 0.73 (0.17)
10 0.41 (0.12) 0.28 (0.13)
30 0.07 (0.02) 0.04 (0.03)
* Values are the means of 10 mussels from each treatment (standard error in
parentheses).
50
-------
4
~r~
50
15-
I0H
£ 8-i
-»
o
co 0-
- 5-
~r~
100
-10-
~
T
0
I
50
~
1.0-1
E 0.8
B
x
| 0.6
ac
o
uj 0.4-
x
(A
0.2-
~r~
100
0.0-
T
0
EXPOSURE CONCENTRATION (% BRH)
Figure 15. Effect of BRH suspended sediment on the clearance rate,
SFG, and shell growth of M. edulis, Day 14, Experiment 1
-------
5-t
•C 4-
UJ
5 *
(C
Ui
o
z
< 2
QC
<
Ui
o
I
~
{
xz
15-
10-
5-
o
u. _
(n 0
- 5-
"i 1—i 1 r~
0 10 20 30 40
-10
{
I
1.0-1
~0.8
E
e
H 0.6
*
o
QC
©
_j 0.4
_i
ui
X
CO
0.2-
i 1 1 1 r o.o-
0 10 20 30 40
1 1 1 1 r
0 10 20 30 40
EXPOSURE CONCENTRATION (% BRH)
Figure 16. Effect of BRH suspended sediment on the clearance rate,
SFG, and shell growth of M. edulis, Day 14, Experiment 2
-------
CO
5-
4-
tii
<
a 3
iu
o
z
-------
91. In Experiment 2 it is also interesting to note the differences
between the first and second 14-day growth periods. The mussels in the
0-percent BRH chamber grew virtually the same amount in the first 14 days as
the second 14. In the 10-percent BRH chamber, growth was reduced in the
second period as compared to the first. This was reflected in the SFG values
as well (Table 13). In the 30-percent BRH treatment, very little growth oc-
curred in either 14-day period.
Residue effects
92. Regression analysis was used to determine whether any relationship
existed between the SFG values and tissue residues. Data from both laboratory
experiments were included because exposure conditions (i.e., temperature and
total suspended particulate levels) were similar in each experiment; only the
percent BRH differed between experiments and treatments. The relationship
between SFG and tissue residue for each of the selected 10 chemicals, and two
summary statistics, are presented graphically in Figures 18-23. The presence
of a regression line on any graph indicates a significant relationship
(P < 0.05) between SFG and the tissue residue for that particular element,
compound, or summary statistic.
93. The individual graphs demonstrate that when a significant relation-
ship exists between SFG and the tissue concentration of a chemical variable,
that relationship is inverse in nature. These findings are consistent with
the results of the residue-effects data presented above and support the con-
tention that mussel residues accurately reflect BRH exposure. From the lab-
oratory data for exposure, residue, and SFG, it is clear that increased
exposure to BRH material is associated with increased tissue residue concen-
trations, reduced SFG, reduced clearance rates, and reduced growth in mussels.
94. Several chemical variables did not relate significantly to SFG val-
ues. Iron residues, for example, would not be expected to relate to SFG be-
cause iron concentrations are indicative of exposure to any sediment. Total
suspended sediment levels were similar in all exposure concentrations.
95. The variable CENT showed no visible pattern to this distribution in
mussels after exposure to different concentrations of BRH material, indicating
that mussels may have accumulated only those contaminants which have a narrow
range of octanol water partitioning coefficients (Lake, Hoffman, and Schimmel
1985). The other summary statistic, SUM, did show a significant negative
relationship with respect to SFG. These data suggest that while the mussels
54
-------
15-
10-
5-
o
u.
CO
-5-
10-
A
A
A
1 1 1 1 1 1 1 1 1
0 25 50 75 100 125 150 175 200 225
PHENANTHRENE (ng/g dry)
a. Phenanthrene
SUM OF 178 ALKYL HOMOLOGS (ng/g dry)
b. Sum of 178 alkyl homologs
Figure 18. Relationship between the SFG of M. edulis and the tissue
residue concentrations of phenanthrene and the sum of the 178 alkyl
homologs in the laboratory experiments. The presence of a regres-
sion line indicates a significant (P < 0.05) relationship between
the two variables
55
-------
FLUORANTHENE (ng/g dry)
a. Fluoranthene
BENZO(o)PYRENE (ng/g dry)
b. Benzo(a)pyrene
Figure 19. Relationship between the SFG of M. edulis and the tissue
residue concentrations of fluoranthene and benzo(a)pyrene in the
laboratory experiments. The presence of a regression line indicates
a significant (P < 0.05) relationship between the two variables
56
-------
SUM OF PAHs (ng/g dry)
a. SUM of PAHs
15
10-
5-
o
u.
w 0-
-5-
-10
1 1 1 1 1 1 1 1 I 1
200 205 210 215 220 225 230 235 240 245 250
CENTR0I0 OP PAHs (mol.wt.)
b. CENT of PAHs
Figure 20. Relationship between the SFG of M. edulis and the summary
statistics, SUM and CENT, in the laboratory experiments. The pres-
ence of a regression line indicates a significant (P < 0.05) rela-
tionship between the two variables
57
-------
PCB AS AI2S4 (ng/g dry)
a. PCB as A1254
I 5-A
k
ETHYLAN (ng/g dry)
b. Ethylan
Figure 21. Relationship between the SFG of M. edulis and the tissue
residue concentrations of PCB as A1254 and ethylan in the labora-
tory experiments. The presence of a regression line indicates a
significant (P < 0.05) relationship between the two variables
58
-------
CADMIUM ij/q/q dry)
a. Cadmium
COPPER (^/g/g dry)
b. Copper
Figure 22. Relationship between the SFG of M. edulis and the tissue
residue concentrations of cadmium and copper in the laboratory ex-
periments. The presence of a regression line indicates a signifi-
cant (P < 0.05) relationship between the two variables
59
-------
£
15-
10-
5-
o
u.
/g/g dry)
Chromium
—i
20
15
10-
I 5H
s
(9
li.
co 0
-5-
-10-
-1 1 1—
150 300 450
—I 1 1 1 1
900 (050 1200 1350 1500
600 750
IRON {j/q/q dry)
b. Iron
Figure 23. Relationship between the SFfi of M. edulie and the tissue
residue concentrations of chromium and iron in the laboratory exper-
iments. The presence of a regression line indicates a significant
(P < 0.05) relationship between the two variables
60
-------
were selective as to which PAH compounds they accumulated (based on the CENT
data), increased exposure levels of BRH material resulted in increased tissue
residue concentrations of some PAH compounds (see paragraph 25).
96. There was also no apparent relationship between SFG and phenan-
threne. This lower molecular weight compound was initially accumulated by
mussels on Day 7 (Figure 18), after which the concentration decreased to a
level similar in all three treatments. The fact that exposure remained con-
stant while the phenanthrene tissue concentrations decreased over time may
indicate the ability of the mussel to depurate or metabolize this compound.
Field
Exposure
97. Estimated from residues. The first method used to determine pos-
sible exposure conditions of M. edulis to BRH material in CLIS involved the
use of laboratory-generated relationships between PCB tissue residues and BRH
exposures. There are several assumptions inherent in this process: mussels
provided an integrated measure of exposure during each field deployment; mus-
sels were at equilibrium with background BRH levels in the water column; and
PCBs are a good chemical marker for BRH material. Based on the results of the
laboratory experiments, each of these assumptions seems reasonable.
98. The predicted exposures for each station and collection date dem-
onstrate several spatial and temporal trends (Table 15). Spatially, the data
indicate a trend toward greater exposure near the CNTR station immediately
following disposal. This is evidenced by the elevated exposures at T = 0
(100E > REFS) and T + 2 (400E > 1000E > REFS) towards the disposal mound.
This pattern disappeared by T + 8, where exposures were nearly the same at the
CNTR, 400E, and 1000E stations, with the REFS station being lower than the
other three.
99. Temporally, the estimated BRH exposures decreased with increasing
time from disposal. The maximum exposure occurred at the 400E station at
T + 2. This value ranged between 1.4 and 0.8 mg/H of BRH suspended sediment,
depending on whether the background concentration at REFS was subtracted. By
the next collection, T + 8, the maximum estimated exposure, also at 400E,
decreased to between 0.7 and 0.3 mg/Jt, half that of the previous collection.
61
-------
Table 15
Predicted BRH Suspended Material Sediment Exposure (mg/&)
Required to Achieve the Measured Tissue Residue
Values of Mussels Deployed in CLIS*
Collection Estimated Exposure Range
Cruise
T - 04
T = 0
T + 2
T + 8
T + 12
T + 15
T + 21
T + 27
Station
High Value
Low Value
CNTR
0.37
0.00
400E
0.26
0.00
1000E
0.38
0.00
REFS
0.38
0.00
1000E
1.04
0.56
REFS
0.49
400E
1.39
0. 79
1000E
0.98
0.38
REFS
0.60
CNTR
0.67
0.21
400E
0.71
0.25
1000E
0.60
0.14
REFS
0.46
CNTR
0.61
0.06
400E
0.64
0.09
1000E
0.53
0.00
REFS
0.55
CNTR
0.84
0.31
400E
0.61
0.08
1000E
0.53
CNTR
0.52
0.12
400E
0.66
0.26
1000E
0.55
0.15
REFS
0.40
400E
0.52
0.09
1000E
0.37
0.00
REFS
0.43
(Continued)
* Each estimate was calculated based on laboratory-generated PCB residue-
exposure concentration relationships. The high value was determined from
the actual mussel tissue residue concentration while the low estimate was
calculated after the REFS PCB residue was subtracted from the other stations
during that collection period.
62
-------
Table 15 (Concluded)
Collection Estimated Exposure Range
Cruise Station High Value Low Value
T + 43 CNTR 0.33 0.06
400E 0.31 0.04
REFS 0.27
T + 55 400E 0.52 0.00
1000E 0.42 0.00
1000E 0.47 0.00
REFS 0.53
T + 116 CNTR 0.30 0.00
400E 0.34 0.00
1000E 0.43 0.01
REFS 0.42
63
-------
Subsequent collections indicated a continued decrease to levels similar to
those at the REFS station by T + 12.
100. Exposures estimated from water chemistry data. In addition to the
estimates of BRH exposure based on mussel PCB tissue residues, a second esti-
mate was made using PCB and copper concentrations in whole water samples taken
postdisposal. The data indicate spatial and temporal trends similar to those
obtained from the tissue residue estimates (Table 16).
101. Spatially, the sample collected on 7 June 1983 showed the highest
BRH estimate (based on copper) at the CNTR station, followed by lower concen-
trations at 400E and 1000E stations, and the lowest levels at REFS. The esti-
mate based on PCB concentrations indicated the CNTR station was elevated
compared to the REFS station. The same pattern was observed in both the cop-
per and PCB estimate for 21 July 1983 sample. A decreasing concentration of
BRH material was estimated moving away from the CNTR of the disposal mound.
102. On a temporal scale, the BRH concentrations (copper data) de-
creased by about half from June to July (high estimate). After this collec-
tion, however, the copper-based BRH estimates fluctuated over time, with the
December 1983 and June 1984 values higher than the September 1983 concentra-
tions. This pattern over time may be more reflective of CLIS than of actual
BRH levels because these estimates (high value) were based on the absolute
copper levels at each location. Inspection of the low estimate indicated a
more distinct pattern over the same time period. The BRH levels were highest
immediately after the disposal operation (June 1983) and generally decreased
with increasing time. The low estimate provided here is more a measure of
relative difference between the stations, after background Long Island Sound
concentrations are subtracted (REFS). When trying to discern temporal trends,
this estimate may be more appropriate.
103. The pattern of f$RH exposure based on PCB water concentrations was
very similar to that of copper. The highest value was detected at the CNTR
station in June 1983 and decreased both spatially and temporally with increas-
ing time. In addition, the high estimates did not show the same variability
over time that the copper data did. This may indicate that PCB concentrations
in Long Island Sound were most constant over time, and thus BRH estimates
based on PCB concentrations were less influenced by background fluctuations.
Tissue residues
104. The tissue residue levels for the mussels collected during the
64
-------
Table 16
Predicted BRH Suspended Sediment Exposure (mg/&) Based on PCB
and Copper Whole Water Chemistry Data*
Estimate Using Cu Estimate Using PCB
Date Station High Low High Low
07 Jun 83 CNTR 1.30 0.71 1.05 0.69
400E 1.12 0.53
1000E 1.14 0.55
REFS 0.59 0.00 0.36 0.00
21 Jul 83 CNTR 0.62 0.26 0.19 0.11
400E 0.49 0.13
1000E 0.41 0.05
REFS 0.36 0.00 0.08 0.00
31 Aug 83 CNTR — — 0.17 0.07
400E — — 0.21 0.11
1000E — ~ 0.16 0.06
REFS — — 0.10 0.00
02 Sep 83 CNTR
400E 0.72 0.22
1000E
REFS 0.50 0.00
05 Dec 83 CNTR — « 0.05 0.00
400E 1.13 0.37 0.08 0.00
1000E — -- 0.09 0.00
REFS 0.76 0.00 0.09 0.00
06 Jun 84 CNTR
400E 1.00 0.09
1000E
REFS 0.91 0.00
* Each estimate was calculated through division of the concentration of PCB
or Cu present in the field by the concentration of that material present in
the BRH barrel material (6,910 ng/g and 2,900 Ug/g for PCB and copper,
respectively). The high value was determined from the actual whole water
concentration while the low estimate was calculated after the REFS values
were subtracted from the other stations during that collection period.
65
-------
course of the FVP study are presented graphically in Figures 24-29 for each of
the 12 selected organic, inorganic, and summary statistic chemical contami-
nants. The raw data summarized in these figures are given in Appendix A.
105. Temporally, the PCB, ethylan, and PAH residues increased during
the disposal operation. After completion of the disposal, tissue residues
decreased to concentrations similar to those from predisposal deployments.
The summary statistic, SUM, reflected the same pattern as most of the PAH
compounds.
106. A consistent pattern emerged when the spatial component of the or-
ganic residue data was considered within a sampling date. Mytilus edulis were
deployed only twice during the actual disposal operation, at T + 0 and T + 2.
For the T + 0 collection, only the 1000E and REFS stations were recovered.
The tissue residue concentration for each organic compound was uniformly
higher at the 1000E station than REFS. The T + 2 collection included data
from three stations—400E, 1000E, and at REFS. Once again, a consistent pat-
tern is seen in the residue data with mussels at 400E exhibiting the highest
concentrations for each compound, followed by the 1000E and REFS stations.
After the completion of disposal, the differences in residue concentrations
decreased dramatically between stations.
107. The tissue residue data for metals did not provide as clear a
picture of the disposal operation as the organic residues. In general, metal
residue concentrations increased slightly during the disposal operation, after
which they decreased to levels well below those present during the predisposal
collection (T - 4). Metal concentrations were elevated in M. edulis collected
in October and November 1983 (T + 21, T + 27), well above those present even
during the disposal operation (T + 0, T + 2). These two samples consisted of
organisms that had been deployed at the FVP site for 7 months and 3 months,
respectively. One possible explanation for elevated metals residues may be
that mussels require a longer period of time to reach steady state with re-
spect to metal concentrations. Comparing the organic and metal residue data
from the field suggests that organic tissue residues present a better picture
of the disposal operation at the FVP disposal site.
Scope for growth effects
108. The results of the physiological measurements are summarized in
Table 17. Predisposal data, T - 4, indicated no differences among the physio-
logical responses or the SFG index from each station (Table 17).
66
-------
250 -i
>»
i.
T>
o>
s
o>
c
200-
ui 150-
ui
a;
i
<
z
Ul
X
a.
100-
50
¦ CNTR
¦ 400E
~ I000E
~ REFS
I
l.tfTi.hn,
i~h.iln. ^~h
SAMPLING DATE
T-4 T+0
1 1 1 1 1 1 1 1 1
T+2 T+8 T + 12 T+15 T+21 T+27 T+43 T+55 T+II6
CRUISE NUMBER
a. Phenanthrene
? 2000-1
¦o
CJ>
c
tn
e>
o
_i
o
2
_J
>-
-J
<
(0
2
1500-
1000-
500-
¦
CNTR
¦
400E
~
I000E
~
REFS
4/83 5/83 6/83 7/83 8/83 9/83 10/83 11/83 3/84 6/84 8/85
SAMPLING DATE
T-4 ' T+0 ' T+2 ' T+8 ' T+12 ' T+15 ' T+21 ' T+27 ' T+43 ' 1+55* T+116 *
CRUISE NUMBER
b. 178 alkyl homologs
Figure 24. Concentrations of phenanthrene and the 178 alkyl homologs
in the tissues of M. edulis exposed at the specified FVP stations and
sampling dates
67
-------
¦o
CP
\
a>
z
UJ
z
a.
o I00H
o
N
Z
liJ
ED
50-
a
| CNTR
¦ 400E
~ I000E
~ refs
JMJ
Ik uu rh—^
4/03 5/83 6/83 7/83 8/83 9/83 10/83 11/83 3/84 6/04 0/05
SAMPLING DATE
l 1
T-4 T+0
1 1 1 1 1 1 1 t n
T+2 T+0 T+12 T+15 T+21 T + 27 T + 43 T+55 T + l 16
CRUISE NUMBER
b. Benzo(a)pyrene
Figure 25. Concentrations of fluoranthene and benzo(a)pyrene
in the tissues of M, edulis exposed at the specified FVP sta-
tions and sampling dates
68
-------
9000-1
>»
u
"O
C*
V
£ 6000
<
a.
u.
o
2
3
£/)
3000-
0- -
XL
4/83 5/83 6/83 7/83 8/83 9/83 10/83 11/83 3/84 6/84 8/85
SAMPLING DATE
I 1 1 1 1 1 1 1 1 1 1 l
T-4 T+0 T+2 T+8 T+12 T+15 T+21 T+27 T+43 T+55 T+II6
CRUISE NUMBER
a. SUM of PAHS
2 50-i
o 240
I
<
o
Q
O
a.
UJ
o
230-
220-
210-J
4/83 5/83 6/83 7/83 8/83 9/83 10/83 11/83 3/84 6/84 8/85
SAMPLING 0ATE
1
T-4 T + 0
1 1 1 1 1 -| 1 1 1
T+2 T+8 T+12 T+15 T+21 T+27 T+43 T+55 T+II6
b.
CRUISE NUMBER
CENT of PAHs
Figure 26. Concentrations of the SUM of the PAHs
and CENT in the tissues of M. edulis exposed at
the specified FVP stations and sampling dates
69
-------
1600-1
\ | c.\j\j
o»
m
cvi
< 000
in
<
CO
o
o. 400
o- wm — i— i"* i ii i— ii— i™™
4/83 6/83 6/83 7/83 8/83 9/83 10/83 11/83 3/84 6/84 8/85
SAMPLING DATE
T
1 r
T"
T
T
T
T-4 T+0 T+2 T+8 T+12 T+15 T+21 T+27 T+43 T+55 T+II6
CRUISE NUMBER
a. PCBs as A1254
120
100-
o»
^ 80
c
< 60-
_i
v
I
UJ
40-
20
o- fcrTI t
LI
4/83 ' 5/83 F 6/83 7/83 8/83 9/83 10/83 11 /83 3/84 6/84 8/85
SAMPLING DATE
T-4 T + 0 T+2 T+8 T+12 T+15 T+21 T+27 T+43 T+55 T + II6
CRUISE NUMBER
b. Ethylan
Figure 27. Concentrations of PCBs as A1254 and ethylan in the
tissues of M. eduZis exposed at the specified FVP stations and
sampling dates
70
-------
T
T
T
T
SAMPLING DATE
1 1
T
T
T-4 T + 0 T+2 T+8 T+12 T+15 T+21 T + 27 T+43 T+55 T+II6
CRUISE NUMBER
a. Cadmium
|CNTR
I 400E
~ IOOOE
~ refs
4/83 ' 5/83 ' 6/83^ 7/83 ' 8/83 ' 9/83 ' 10/83' 11/83' 3/84 ' 6/84 ' 8/85
SAMPLING DATE
T-4 ' T+0 ' T+2 ' T+8 ' T+12 ' T+15 ' T+21 ' T+27 ' T+43 ' T+55 ' T+II6
CRUISE NUMBER
b. Copper
Figure 28. Concentrations of cadmium and copper in
the tissues of M. edulis exposed at the specified
FVP stations and sampling dates
71
-------
i i i r r
4/83 5/83 6/83 7/83 8/83 9/83 10/83 11/83 3/84 6/84 8/85
SAMPLING DATE
I 1 1 1 1 1 1 1 1 1 1— 1
T-4 T + 0 T+2 T + 8 T+12 T+15 T+21 T+27 T+43 T+55 T+II6
CRUISE NUMBER
a. Chromium
1500-1
•ST iooo
o>
z
o
tc
500
0 "T
4/83 5/83 6/83 7/83 8/83 9/83 10/83 11/83 3/84 6/84 8/85
SAMPLING DATE
' T-4 ' T+0 ' T+2 ' T + 8 ' T+12 ' T+15 ' T+21 ' T + 27 ' T+43 ' T+55 ' T+U6
CRUISE NUMBER
b. Iron
Figure 29. Concentrations of chromium and iron in
the tissues of M. edulis exposed at the specified
FVP stations and sampling dates
72
-------
Table 17
Mean Values* (N = 10) of the Physiological Parameters Measured on Mussels
from CLIS Collections with the Mean SFG Value Calculated for Station
Station
Clearance
Rate
4/hr
Absorption
Efficiency
percent
Respiration
Rate
ml O^/hr
Ammonia
Excretion
Ug NH^/hr
Scope for
Growth
J/hr
T -
4, 22 Apr 83
CNTR
2.35
(0.31)
94
(0.4)
0.61 (0.03)
23.08 (2.08)
3.33
(1.53)
400E
2.81
(0.25)
94
(0.5)
0.65 (0.02)
26.64 (1.28)
4.63
(1.45)
1000E
2.33
(0.21)
96
(0.4)
0.67 (0.04)
31.11 (4.00
2.31
(1.20)
REFS
2.20
(0.34)
95
(0.3)
0.66 (0.06)
21.51 (3.07)
1.36
(1.47)
T +
0, 24 May 83
1000E
4.88
(0.64)
92
(0.7)
0.72 (0.06)
16.51 (2.16)
9.46
(2.16)
REFS
5.05
(0.62)
91
(0.9)
0.73 (0.04)
13.99 (1.87)
8.87
(1.71)
T +
2, 07 Jun 83
400E
6.32
(0.54)
95
(0.4)
0.87 (0.04)
8.78 (1.26)
11.21
(1.76)
1000E
5.01
(0.58)
96
(0.7)
0.90 (0.05)
7.62 (2.17)
6.50
(1.38)
T +
8, 13 Jul 83
CNTR
3.53
(0.36)
88
(0.6)
0.95 (0.07)
20.19 (3.79)
-0.11
(1.18)
400E
4.63
(0.56)
89
(0.7)
0.84 (0.04)
21.81 (1.69)
5.04
(1.51)
1000E
5.69
(0.58)
83
(0.9)
0.91 (0.08)
19.80 (3.28)
6.30
(1.02)
REFS
5.39
(0.66)
85
(0.7)
0.80 (0.02)
14.08 (1.90)
6.85
(2.37)
T +
12, 10 Auk 83
CNTR
3.48
(0.31)
91
(1.1)
0.92 (0.16)
16.73 (3.28)
0.93
(1.47)
400E
4.02
(0.68)
94
(0.8)
0.95 (0.23)
14.40 (3.32)
2.73
(2.37)
1000E
3.43
(0.51)
96
(0.4)
0.97 (0.18)
40.35 (4.45)
0.18
(2.22)
REFS
3.65
(0.54)
96
(0.2)
0.88 (0.21)
34.19 (3.88)
2.97
(1.95)
T +
15, 06 Sep 83)
CNTR
3.78
(0.56)
92
(2.7)
0.77 (0.09)
33.85 (8.35)
4.70
(1.75)
400E
3.64
(0.44)
88
(4.7)
0.76 (0.04)
18.83 (2.91)
3.94
(1.74)
1000E
3.41
(0.57)
95
(4.8)
0.76 (0.06)
21.46 (2.92)
4.86
(2.77)
REFS
3.31
(0.35)
85
(4.6)
0.82 (0.06)
25.40 (1.88)
1.47
(1.74)
(Continued)
Note: Missing values during a collection period indicate the station was
lost.
* Values in parentheses are standard errors.
73
-------
Table 17 (Concluded)
Station
CNTR
400E
1000E
REFS
400E
1000E
REFS
CNTR
400E
REFS
400E
1000E
REFS
400E
1000E
REFS
CNTR
400E
1000E
REFS
Clearance
Rate
£/hr
1.91 (0.14)
1.69 (0.28)
1.96 (0.28)
2.99 (0.43)
1.78 (0.31)
2.06 (0.37)
1.63 (0.45)
0.11 (0.05)
1.17 (0.19)
1.58 (0.27)
3.77 (0.76)
4.37 (0.57)
4.49 (0.40)
2.52 (0.41)
3.55 (0.64)
3.55 (0.63)
0.81 (0.12)
1.18 (0.19)
1.39 (0.37)
0.73 (0.08)
Absorption
Efficiency
percent
Respiration
Rate
ml 02/hr
91
89
88
88
T + 21, 18 Oct 83
(0.6)
(2.7)
(0.6)
(1.5)
0.74
0.66
0.59
(0.05)
(0.06)
(0.04)
0.63 (0.05)
T + 27, 29 Nov 83
96 (0.3)
96 (0.4)
95 (0.5)
0.42 (0.04)
0.52 (0.05)
0.37 (0.05)
T +43. 20 Mar 84
76 (6.1)
94 (0.6)
95 (0.4)
0.42 (0.03)
0.46 (0.03)
0.55 (0.09)
T +55. 05 Jun 84
89 (0.4)
79 (1.7)
89 (0.7)
0.73 (0.05)
0.74 (0.07)
0.92 (0.05)
T + 74, 17 Oct 84
76 (4.3)
75 (4.2)
77 (2.4)
0.48 (0.05)
0.54 (0.03)
0.51 (0.04)
T + 116, 14 Aug 85
80 (1.0)
80 (0.5)
78 (0.7)
82 (1.6)
0.49 (0.03)
0.54 (0.04)
0.68 (0.07)
0.57 (0.06)
Ammonia
Excretion
yg NH^/hr
27.18 (2.53)
28.35 (3.81)
24.94 (1.75)
25.83 (2.78)
24.65 (1.30)
26.39 (2.35)
23.47 (2.59)
33.94 (2.50)
23.56 (2.50)
46.85 (9.62)
22.94 (2.88)
9.18 (1.33)
24.30 (2.89)
23.98 (2.58)
37.31 (2.45)
23.31 (3.39)
18.39 (2.81)
13.65 (2.65)
14.32 (3.43)
19.86 (3.78)
Scope for
Growth
J/hr
-1.38 (0.98)
-2.49 (1.81)
0.29 (1.76)
3.75 (1.54)
4.53 (2.10)
3.08 (1.74)
2.71 (2.92)
-8.50 (0.55)
-0.54 (1.42)
0.40 (1.20)
5.53 (2.61)
3.00 (2.31)
3.87 (1.31)
1.65 (1.48)
5.27 (2.43)
6.29 (1.94)
-4.24 (0.85)
-3.01 (1.15)
-4.65 (1.55)
-6.02 (1.27)
74
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109. Mussels collected at T + 0 and T + 2 had been deployed at the CLIS
site for 1 month and 6 weeks, respectively. Differences in ammonia excretion
rates were observed. However, the other physiological variables, as well as
SFG values, were not different among stations. Loss of the 400E and REFS sta-
tions for the T + 0 and T + 2 collections, respectively, was unfortunate.
This did not permit a comparison between the stations that presumably were
most impacted (400E) and least impacted (REFS) during this time period.
110. A reduction in mean clearance rate was noted at the CNTR station
compared with the rates at the 1000E and REFS stations during the T + 8 col-
lection (Table 17), resulting in a subsequent reduction in the mean SFG of the
CNTR mussels.
111. The SFG differences observed at the T + 8 collection were not
present 1 month later in the T + 12 collection (Table 17). Mussels deployed
for this monthly period exhibited higher mean ammonia excretion rates at the
1000E and REFS stations; however, there were no corresponding differences in
the mean SFG index between stations.
112. No differences were observed among either the individual physio-
logical parameters or the SFG index at the four stations at the T + 15 collec-
tion (Table 17).
113. The physiological measurements of the mussels collected at T + 21,
after a 6-month deployment (Table 17), indicated a difference in clearance
rates and SFG among stations. Mussels from the REFS station had a higher mean
clearance rate than the mussels from the other three stations. The SFG data
indicate a lower mean SFG at the CNTR and 400E stations compared with the REFS
station, with the mean SFG of the 1000E mussels not different from the other
three stations.
114. The T + 27 mussel collection was completed after a 3-month deploy-
ment at the FVP disposal site. An increase in the mean respiration rate was
noted in the mussels collected from the 1000E station (Table 17). No differ-
ences were present in the mean SFG values among the stations.
115. The physiological data for the mussels collected at T + 43, after
a 3-month deployment (Table 17), showed that mean clearance rate, mean ab-
sorption efficiency, and mean SFG were lower in mussels from the CNTR station
compared with those of the mussels from the other two stations. The mean
clearance rate of the CNTR mussels was almost zero. In addition, the mean ab-
sorption efficiency was lower than the other stations. Small differences in
75
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absorption efficiency of 1 to 5 percent may not be biologically significant
due to measurement variability. In this instance, however, the mean absorp-
tion efficiency of mussels from the CNTR station was almost 20 percent lower.
Finally, the mean SFG response of the mussels from the CNTR station was lower
than the mussels at the 400E and REFS stations.
116. The mussel collection at T + 55 represented a deployment period of
8 months (Table 17). There were no differences between the mean SFG values
for any of these stations; however, mussels retrieved from the 1000E station
exhibited lower mean absorption efficiencies and lower mean ammonia excretion
rates than mussels from the 400E and REFS stations.
117. Mussels retrieved during the T + 74 collection had been deployed
for a period of 4 months (Table 17). Mean ammonia excretion rates were higher
in mussels from the 1000E station than those of the mussels from the 400E and
REFS stations. There were no differences among the mean SFG values from the
three stations.
118. The final collection from CLIS, T + 116, was completed in August
1985. Mussels were deployed for a period of 1 month to observe whether there
were any long-term effects from the disposal of the BRH material. The data
indicate no differences among stations for any of the physiological mea-
surements (Table 17); however, the SFG values were low at all stations.
Residue-effects data
119. The relationship between SFG effects and tissue residues measured
in the field mussels was not as clear as it was for the laboratory experiments.
The SFG of a mussel is influenced by a variety of extrinsic (i.e., tempera-
ture) and intrinsic (i.e, gametogenesis) factors which varied naturally from
collection to collection during the FVP. These normal seasonal changes in SFG
preclude regressing all of the field SFG and tissue residue data together be-
cause they would conceal any relationship that might exist. Therefore, the
field residue-effects data will be considered within discrete sampling peri-
ods, where mussels were presumably exposed to similar temperatures and should
be in a similar stage of gametogenesis. As was stated at the beginning of
this report, the purpose for using the SFG index was to determine whether
relative sublethal effects could be measured between laboratory treatments or
field stations. Consideration of field SFG-residue relationships within a
sampling period is entirely consistent with this objective.
120. The use of SFG values within a sampling period reduced sample 3ize
76
-------
to a maximum of four, when all stations were recovered. Regression analysis
with this sample size is not appropriate; therefore, the data are presented
graphically to illustrate trends. It would be impractical to present graphs
for all of the residue-effects data for each of the 12 sampling dates. PCBs
were selected because of the good relationship previously described between
residues of this compound and SFG in the laboratory experiments, as well as
the relationship between BRH exposure concentration and mussel PCB residues in
those same laboratory experiments. Therefore, only the field residue-effect
data for PCBs and SFG will be presented here. A complete comparison of
residue-effects data for the field collections can be made by comparing field
SFG values (Table 17) with field mussel residues (Appendix A, Tables A3-A13).
121. The SFG-PCB tissue residue data are presented in Figure 30. Mus-
sels collected predisposal showed a very narrow range of PCB residues with a
corresponding small range in SFG values. The T = 0 collection indicated that
mussels from 1000E exhibited elevated PCB residues compared with REFS; how-
ever, SFG values from both stations were similar. Mussels collected at T + 2
once again displayed higher residues closer to the disposal mound, 400E com-
pared with 1000E, while SFG values were not that different from each other.
Subsequent field collections indicated a reduction in PCB tissue residues from
all stations. At T + 8, mussels from the CNTR station had a lower SFG value
compared with those at the other three stations; however, there was no corre-
sponding difference in PCB residues at this time. Mussels from the T + 12,
T + 15, T + 21, T + 27, T + 55, and T + 116 collections exhibited a very small
range of PCB tissue residues and a corresponding narrow range between SFG
values at the four stations. The only exception was the T + 43 collection
where a reduction in SFG at the CNTR station was not related to any differ-
ences in PCB residues among stations.
122, These data indicate that no distinct relationship was evident be-
tween SFG and tissue residues in the field. Several possible explanations
will be put forth in the discussion; however, one fact will be mentioned here.
The highest mussel PCB residue concentration in the field (T + 2, 400E) was
less than the lowest tissue residue in the laboratory experiments (10 percent,
Day 28). These data suggest that residue concentrations of field-exposed mus-
sels may have been too low to elicit a SFG effect.
77
-------
15 n
10
5
0-
-5-
A 0% BRH
A 10% BRH
& 30% BRH
-10-
15-
10-
o _
h. 0-
V)
-5-
-10-
15-
10-
5H
0-
-5-
-10-
a. Laboratory
o
%
¦ i i i i
O CNTR
• 400E
~ I000E
¦ REFS
b. T-4,22 Apr 83
i i i i i i
500 1000 1500 2000 2500 3000 3500 4000
PCS AS AI254 (ng/gdry)
c. T»0, 24 MAY 83
Figure 30. Relationship between the SFG of M. edulis and PCB
tissue residue concentrations in laboratory and field-exposed
animals. The laboratory data are presented to provide a per-
spective between the residue concentrations of laboratory and
field-exposed mussels (Sheet 1 of 4)
78
-------
O CNTR
• 400E
~ I000E
¦ REFS
d. T + 2, 07 JUN 83
-5-
-10-
is-
le-
s'
0-
-5-1
-10*
i i t i i i i i
•. Tt8, 13 JUL 83
i i i i ¦ i i i
500 1000 1500 2000 2500 3000 3500 4000
PCB AS A1254 (ng/g dry)
f. T + 12, 10 AUG 83
Figure 30. (Sheet 2 of A)
79
-------
I51
10-
5-
0-
a o
O CNTR
• 400E
a I000E
¦ REFS
T I I
g. T + 15, 06 SEP 83
a
o
i i i i
h. T-f 21, 18 OCT 83
i i i i i i i i
500 1000 1500 2000 2500 3000 3500 4000
PC8 AS AI254 (ng/q dry)
i. T +27, 29 NOV 83
Figure 30. (Sheet 3 of 4)
80
-------
15-
10-
O CNTR
• 400E
~ I000E
¦ REFS
OH
-5-
-10-
15-
10-
IA
-5-
]. T + 43, 20 MAR 84
HO-
15-
t i i
k. T +55, 05 JUN 84
10-
5-
0-
-5-
-10-
O o
I I I I I I I I
500 1000 1500 2000 2500 3000 3500 4000
PC8 AS AI254 (ng/g dry)
I. T+ 116, 14 AUG 85
Figure 30. (Sheet 4 of 4)
81
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Laboratory-to-Fleld Comparison
123. The laboratory-to-field comparison was completed in two parts and
included both tissue residue and effects data. The approach taken was to
first establish when exposure conditions were similar in the laboratory and
the field by comparing laboratory and field residue data. Comparable tissue
residues were interpreted as being indicative of comparable BRH exposures.
The second step was to compare the SFG values of the laboratory- and field-
exposed mussels with similar tissue residue concentrations.
Residues
124. Results of the cluster analysis suggested several general observa-
tions. First, the samples that were most similar included all the field resi-
dues collected after T + 2 and the laboratory 0-percent BRH exposures. This
would indicate that mussels in the field received minimal exposure to BRH ma-
terial after the initial disposal operation. Second, mussels collected pre-
disposal (CNTR, 400E, 1000E) and those collected shortly after disposal (1000E
at T « 0 and T + 2) were more similar to the other field samples than the lab-
oratory samples. This would imply that even during disposal, BRH exposures at
these stations were more similar to subsequent postdisposal field residues
than to laboratory BRH exposures. Third, all of the laboratory residues were
more similar to each other than to any of the field samples. This grouping
would indicate that all laboratory exposures were very different from field
exposures. Finally, mussel residues obtained from 400E at T + 2 were more
similar to those of the laboratory-exposed mussels than to the other field ex-
posures. This sample was the last to cluster, indicating that it was not very
similar to any other samples; however, it was more closely related to the lab-
oratory samples than to the field samples.
Effects
125. Analysis of the residue data suggested that the most valid com-
parison between laboratory and field SFG data would be between field samples,
with the exception of 400E at T + 2, and laboratory mussels exposed to
0-percent BRH. Comparison of the SFG of mussels exposed to even 10-percent
BRH in the laboratory to any mussels in the field would not be appropriate be-
cause the residues were dissimilar. Additionally, comparison of mussels col-
lected from the field when environmental conditions were not similar to those
in the laboratory would not be appropriate.
82
-------
126. Laboratory experiments were conducted in the spring at water tem-
peratures of 15° C. Comparable field conditions existed at T = 0 and T + 2 in
the field. The SFG value of mussels exposed to 0-percent BRH in the lab for
28 days was 7.2 J/hr. The field-exposed mussels exhibited SFG values of 9.5,
8.9, and 6.5 J/hr, respectively, for the T = 0 (1000E, REFS) and T + 2 (1000E)
collections. The SFG values were very similar, indicating that the relative
physiological conditions of these mussels were the same when environmental
conditions were similar. The only other collection that occurred in the
spring when water temperatures were similar was at T + 55; however, these mus-
sels had been deployed in CLIS for 8 months. The SFG values of these mussels,
3.8, 3.5, and 3.5 J/hr, were slightly lower than that in the 1-month labora-
tory exposure (7.7 J/hr).
127. Several generalizations are apparent from the comparison of labo-
ratory and field results. The laboratory exposures indicated a good relation-
ship between BRH exposure and residue concentrations in M, edulis, These data
provide justification for assuming that lower residue concentrations in the
field-exposed mussels were indicative of lower BRH exposures in the field. In
fact, the highest field residues were less than the residues of the mussels
exposed to the lowest BRH concentration (1.5 mg/£) in the laboratory. There-
fore, the data suggest that there was little or no overlap in laboratory and
field BRH exposure concentrations. The resultant effects data indicated dra-
matic adverse effects on mussels even at the lowest laboratory exposure con-
centration whereas, in the field, few if any effects were attributable to BRH
exposure.
83
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PART IV: DISCUSSION
128. The objectives of this study were to: (a) investigate the
residue-effect relationships in the mussel after independent laboratory and
field BRH exposures and (b) field verify the laboratory results. The design
of this study followed a logical progression from BRH exposure to tissue resi-
due concentration to biological effects. The discussion will parallel this
approach by establishing the exposure-residue and residue-effect relationships
separately in the laboratory and in the field. Finally, a comparison of the
laboratory and field results will be made.
Laboratory Experiments
129. There was a strong link between exposure to BRH sediment and
subsequent tissue residues in M. edulis, as confirmed by the monitoring data
collected during the laboratory experiments. In addition, the relationship
between mussel tissue residues for several chemical contaminants demonstrated
that compounds with higher molecular weights and stability, PCBs in particu-
lar, tracked the BRH exposure concentrations remarkably well. For example,
tissue residue data indicated that mussels from the 30-percent BRH chamber ex-
hibited twice the level of PCBs as those in the 10-percent BRH chamber. Cor-
responding monitoring data indicated that the actual delivered level of BRH
sediment was 3.3 and 1.5 mg/£, respectively, for those two chambers, indicat-
ing that PCBs were a good "marker" for exposure to BRH material. Because of
this direct relationship, residue concentrations can be assumed to be indica-
tive of exposure concentration for highly stable compounds such as PCB. This
relationship is particularly important in the field where direct, continuous
monitoring data of exposure conditions are difficult, if not impossible, to
collect.
130. In addition to the strong exposure-residue relationship, the lab-
oratory experiments indicated a good relationship between mussel residue con-
centrations and biological effects. The residue-effects data presented in
this study were never intended to determine cause and effect. However,
relationships between individual contaminants and biological effects present
strong evidence that observed reductions in scope for growth, clearance rate,
and shell growth were related to exposure to BRH material.
84
-------
131. Mussel SFG values were reduced after exposure to BRH dredged mate-
rial on Day 14 of the first experiment and both Days 14 and 28 of the second
experiment. While there is no attempt to attribute observed decreases in SFG
in the present experiments exclusively to any one contaminant in the BRH sedi-
ment, there is evidence to suggest that some of the contaminants are capable
of causing the observed SFG reductions. Stickle et al. (1985) reported an in-
verse relationship between SFG of mussels and water-soluble fraction aromatic
hydrocarbon concentrations. Widdows et al. (1982) have demonstrated reduc-
tions in the SFG of mussels exposed to the water-accommodated fraction of
North Sea oil. Gilfillan (1975) reported a net decrease in carbon flux in
M. edulis after exposure to crude oil extracts. Copper was also found to re-
duce the SFG of M. edulis (Moore et al. 1984).
132. In addition to the fact that BRH material affected SFG, the shape
of the dose-response curve at Day 14 is also of interest (Figure 14). This
relationship, best described by a curvilinear function, implies that exposure
to some BRH concentration less than 3.3 mg/I could have no effect on SFG in
mussels, i.e., a possible "threshold" concentration of BRH material is re-
quired to cause adverse physiological effects. A similar trend was displayed
by the Day 28 data where a relatively small SFG difference was found between
mussels exposed to 1.5 mg/JL and 3.3 mg/I (0.14 and -1.8 J/hr, respectively)
compared with mussels exposed to no BRH material (7.16 J/hr).
133. Inspection of the individual physiological parameters indicated
that reductions in SFG may be related exclusively to decreased clearance
(feeding) rates. Absorption efficiencies, respiration rates, and ammonia
excretion rates were not different among treatments at Day 14 or Day 28
(Tables 10-12). The impact of BRH suspended sediment on clearance rates was
consistent in both experiments and almost identical to that between SFG and
BRH levels (Figures 15-17). This type of response was observed by Stickle
et al. (1985) and Widdows et al. (1982) in M. edulis after exposure to oil ex-
tracts. Gonzales et al; (1979) reported reduced clearance rates in M. edulis
after exposure to No. 2 heating oil. Nelson, Black, and Phelps (1985) found
lower clearance rates in mussels after exposure to anaerobic BRH dredged ma-
terial. Reductions in clearance rate have been observed in other species as
well. Gilfillan et al. (1976) reported a reduction in filtration in the soft-
shelled clam Mya arenaria from areas exposed to oil spills. Stickle, Rice,
and Moles (1984) found that reductions in the SFG of the gastropod Thais lima
85
-------
after exposure to hydrocarbons were primarily due to reduced feeding rates.
134. Clearance rate measurements in the exposure system provide fur-
ther evidence that BRH suspended sediment affected clearance rates. Measure-
ments on Day 9 in the first experiment (Table 8) demonstrated a dramatic
reduction in the clearance rates of mussels from the 50- and 100-percent BRH
treatments. Measurements in the second experiment, on Days 7 and 16, indi-
cated that exposure to as little as 1.5 mg/i BRH for 7 days was sufficient to
produce dramatic clearance rate reductions. There was the possibility, ini-
tially, that the reduced clearance rates in the exposure chambers represented
a behavioral response (i.e., the mussels reduced their clearance rates in re-
sponse to the BRH material). However, the SFG measurements were made in the
absence of BRH material. Therefore, the differences observed in the clearance
rates of mussels from the exposure system were not due to behavior. Rather,
reductions in clearance rates were probably due to physiological impairment
from the BRH material.
135. One explanation for the reduced clearance rates has been suggested
by histopathological observations. Mussels from treatments containing BRH
suspended sediment showed a loss of cilia from the gill filaments, while those
exposed to REFS sediment alone were normal (Yevich et al. 1986). This infor-
mation is consistent with clearance rate being the only parameter altered in
the BRH treatments because the coordinated movement of these cilia is respon-
sible for creating the currents necessary to move particles into and out of
the mussel. The present study augments the evidence in the literature that
clearance rates are particularly sensitive to pollutants of the type found in
the BRH dredged material.
136. Shell growth data (Table 14 and Figures 15-17) supported the
relative effects of BRH sediment shown by the SFG index. Actual growth obser-
vations could be explained on the basis of the SFG and clearance rate measure-
ments completed over this same time period. Reduced clearance rates in the
treatments containing BRH sediment resulted in decreased energy consumed by
those mussels. These reductions occurred in as little as 7 to 9 days, evi-
denced by the data in Table 8. The adverse effect of the BRH sediment was
observed in the physiological measurements on Day 14. Reduced SFG values in-
dicated that relatively less energy was available for growth with increasing
concentration of BRH, which is exactly what the growth data reflected. Expo-
sure to BRH sediment for an additional 14 days produced continued low SFG
86
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values in the 10- and 30-percent BRH treatments, and subsequent lower growth
in these mussels over the second 14-day period.
137. Agreement between the SFG index and shell growth has been reported
by Bayne and Worrall (1980) for mussels and Gilfillan and Vandermeulen (1978)
in M. arenaria. The close correspondence between the SFG response and shell
growth in the present study supports the use of standardized conditions to
measure relative sublethal effects in mussels. The measurement of SFG under
standardized conditions is sometimes criticized as being nonreflective of what
occurred under field exposure conditions. The results of this study indicated
that the relative differences in SFG values were indicative of actual changes
in shell growth.
138. The similarity between SFG and actual growth raises the obvious
question: why not measure shell growth alone and omit SFG altogether? The
SFG index reflects the total energy available for both somatic growth and re-
production. Energy available for somatic growth can be further partitioned
into either tissue growth or shell growth. Changes in shell length alone
quantify only one component of the organisms' response to the environment,
while the SFG index quantifies the total energetic response. The ultimate
question in pollution biomonitoring is whether or not changes in biological
effects measurements on individuals reflect changes in populations and commu-
nities. Reductions in SFG have been correlated with a decrease in reproduc-
tive output of individual mussels, and, therefore, may be indicative of
broader ecological consequences as well (Bayne, Clark, and Moore 1981).
Changes in shell growth alone, while important, may not provide as much
information about possible ecological effects as SFG. Whenever possible, both
SFG and actual growth should be measured concurrently to provide as much
biological effects data as possible.
139. To summarize the laboratory studies, tissue residues in M. edulis
were directly related to exposure concentration of BRH material. PCBs, in
particular, were a good marker for BRH suspended sediment exposure concentra-
tion. A consistent, inverse relationship was observed between the SFG, clear-
ance rate, and shell growth of mussels and increased exposure to BRH suspended
sediment. Relative differences in SFG were supported by similar changes in
shell growth. Based on these data, a BRH suspended sediment concentration
equal to 1.5 mg/i. would be predicted to cause a similar effect in the field.
87
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Field Experiments
140. The exposure-residue relationships generated in the field portion
of this study were not as straightforward as those described for the labora-
tory studies. While the laboratory experiments provided as constant a set of
exposure conditions as possible, the data required for defining comparable ex-
posure conditions in the field were difficult, if not impossible, to collect.
Consequently, the exposure-residue relationships established in the field
were, by necessity, more qualitative than quantitative.
141. Although qualitative, estimates of BRH exposure in the field were
necessary to establish exposure conditions for the laboratory-to-field compar-
ison (i.e., establish when exposure conditions were similar in the laboratory
and the field). The following generalizations are evident from the estimated
BRH exposure concentrations: (a) the two independent methods, tissue residue
and whole water analysis, provided remarkably similar estimates of BRH expo-
sure and (b) there was a distinct exposure signal at 1 m above the bottom dur-
ing and immediately postdisposal and that signal was transient, decreasing
spatially and temporally postdisposal.
142. Comparison of the tissue residue and water chemistry estimates of
BRH concentrations in CLIS indicated very good correspondence between the two.
Several examples demonstrate this point. Tissue residue data (PCBs) from the
T + 2 collection (Table 15) indicated that the BRH concentration was estimated
to range between 1.4 and 0.8 mg/Jt at the CNTR station. Water samples from the
same station estimated the BRH concentration to range between 1.1 and 0.7 mg/S,
(Table 16) using PCB values, and 1.3 and 0.7 using copper values. Approxi-
mately 1 month later, BRH concentration estimates, based on PCB tissue resi-
dues, were between 0.7 and 0.2 mg/Jl at the CNTR station (T + 8, Table 15).
The corresponding water chemistry estimates of BRH concentrations at the CNTR
station ranged from 0.2 to 0.1 mg/Jt, using PCBs, and 0.6 to 0.3 mg/£, using
copper. The similarity of these estimates indicates that exposure based on
whole water chemistry concentrations and tissue residue concentrations tracked
reasonably well.
143. The good relationship between the residue and water chemistry BRH
estimates further supports the second generalization mentioned above: BRH
exposure 1 m above the bottom was maximal immediately postdisposal and
decreased over time. Spatially, both the PCB tissue residue and water
88
-------
chemistry data indicated that BRH exposure decreased moving away from the CNTR
station immediately postdisposal (Tables 15 and 16). This pattern persisted
until T + 12, when tissue residues were similar at each station. The loss of
the spatial differences in BRH exposures would suggest that exposure from the
disposal mound was minimal after this collection period. Temporally, the
maximum estimated concentration of BRH material 1 m above the bottom ranged
between 1.4 and 0.8 mg/£ (tissue residues at T + 2, Table 15). At the time of
the next collection this value decreased by approximately one-half, and con-
tinued to decrease over time.
144. While a range was calculated to estimate BRH exposure concentra-
tions, it is interesting to note the temporal pattern of the high and low es-
timates. The high estimate of BRH exposure never reached zero, even in the
later collections (i.e., T + 55, T + 116). This may indicate that background
PCB levels in CLIS contributed to the BRH estimates, including those immedi-
ately postdisposal. The low estimate, calculated by subtracting the
concentration at the REFS station, was assumed to remove the background con-
centration present in CLIS. Therefore, the low estimate, while providing a
measure of relative difference between the stations, also may have provided
the best estimate of actual BRH concentration. However, even using the high
estimates, the data suggest that the integrated exposure of BRH material to
M. edulis, 1 m above the bottom, was minimal at all the FVP stations and de-
creased rapidly following completion of the disposal operation.
145. The relationship between residue and effect in the field was not
as direct as that in the laboratory. Several apparently contradictory results
occurred between the residue data and the SFG data in the field. First, dur-
ing the T + 2 collection, when maximum residues (i.e., exposures) occurred, no
decrease in SFG was noted. Secondly, mussels retrieved from the CNTR station
at T + 8, when residue concentrations were lower than at T + 2, did appear to
exhibit an adverse SFG effect. These results suggested several alternative
conclusions: (a) the highest field BRH exposures had no adverse effect on SFG
in mussels, (b) an effect may have been observed if samples were not lost,
(c) an "overshoot" occurred in the physiological response of the T + 2 mus-
sels, or (d) temperature differences between collections may have caused dif-
ferential sensitivity to BRH material.
146. First, it is possible that the BRH exposure concentration at T + 2
may have been insufficient to cause a reduction in SFG. The estimated maximum
89
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BRH exposure at a station where mussels were deployed was between 1.4 and
0.8 mg/£. If the lower estimate (0.8) for exposure was more accurate, it
would represent about half that of the lowest suspended sediment concentration
in the laboratory (1.5 mg/£). Therefore, the signal in the field may have
been "weaker" than that present in the laboratory experiments. This field
level may have been below a possible threshold concentration required to
elicit a physiological response, as suggested by the laboratory experiments.
147. Another possibility concerns the fact that the T + 8 collection
was the first in which mussels were recovered from all four stations. Com-
parison of mussel SFG values from the REF station at T + 2, lost prior to bio-
logical sampling, with those from the 400E station would have presumably
represented the widest range of residue concentrations from the field. It is
possible that some biological effect may have been observed between these two
stations; that, however, is only speculation. Future studies will include
greater redundancy with respect to mussel deployments in order to minimize
sample losses.
148. There is the possibility that a physiological "overshoot" occurred
in the T + 2 mussels as a result of their removal from CLIS to clean water in
the laboratory. Widdows, Donkin, and Evans (1985) reported that the SFG of
mussels removed from chronic oil exposures returned to levels greater than
control mussels. This reported physiological "overshoot" was greater in mus-
sels exposed to higher concentrations of oil than to lower concentrations.
While this phenomenon would account for the elevated SFG of the mussels at
400E, the time frame in the present study makes this unlikely. Widdows,
Donkin, and Evans (1985) indicated that this overshoot occurred between 10 and
20 days after removal from the oil exposures. In the present study, SFG mea-
surements were initiated on field mussels within 24 hr of collection from the
field. Therefore, total recovery from field exposure to BRH in this short
time period would be unlikely. Previous experiments involving field exposure
of M. eduli.8 to sewage sludge indicated that reduced SFG values persisted in
mussels 7 days after they were returned to clean laboratory seawater.*
149. The most plausible explanation for the SFG results observed in
mussels collected at T + 2 and T + 8 concerns possible differential effects
due to temperature. Bayne (1976) reported that the SFG of M. edulis remains
* Unpublished data, William Nelson, SAIC, Narragansett, R. I.
90
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relatively independent of temperature from 10° to 20° C. Above this range,
physiological mechanisms responsible for metabolic compensation begin to break
down. It is possible that the mussels collected at T + 8, when water tempera-
tures were approximately 20° C, were less able to compensate for these condi-
tions and thus exhibited an increased sensitivity to BRH material. Mussels at
T + 2, when water temperature was 15° C, indicated no adverse SFG effect.
These mussels may have been able to compensate physiologically for the
slightly higher BRH concentrations.
150. A second point is that the clearance rates in the mussels from the
CNTR station at T + 8 were reduced only slightly compared to mussels at the
other three stations. In the laboratory, mussels exposed to 1.5 mg/£ for
28 days exhibited dramatic reductions in clearance rates compared to mussels
exposed only to REF material. This estimated concentration of BRH in the
field (0.7-0.2 mg/&) may have been near the threshold level required to affect
the mussels.
151. To summarize, these two collections, mussels retrieved at T + 2
were unaffected by the estimated concentration of BRH suspended sediment pres-
ent during this time period because this concentration may have been insuffi-
cient to cause a negative effect. Mussels collected at T + 8, while exposed
to lower BRH concentrations, were slightly affected by the BRH material, as
evidenced by lower clearance rates and SFG. Elevated temperatures (20° C)
during this collection most likely increased the sensitivity of the mussels to
BRH material.
152. After the T + 8 collection, tissue residue levels of PCB, PAH, and
ethylan decreased with time for the subsequent monthly collections. In addi-
tion, SFG values were similar at all four stations during these collections.
It would appear that these reduced residues did not adversely affect SFG even
with the increased water temperatures during these collections (approximately
20° C). These data are consistent with the suggestion that most field concen-
trations of BRH suspended sediment were below that required to elicit a physi-
ological response, and that the BRH concentrations present at the CNTR station
at T + 8 (0.7-0.2 tag/I) may have been very close to the suggested threshold
concentration.
153. In addition to 1-month field deployments, other mussels were de-
ployed for longer periods of time. Interpretation of these physiological data
was more difficult than for the 1-month deployments. It followed that if it
91
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was difficult to adequately explain exposures during the 1-month deployments,
exposures of 3 months (T + 27, T + 43, T + 74) or 7 months (T + 21, T + 55)
would increase the probability that even more unanticipated, unaccountable
events might occur. For example, mussels collected at T + 21 were deployed in
CLIS for 7 months. Mussels from REF exhibited a higher clearance rate than
mussels from the other three stations. In addition, SFG values at the CNTR
and 400E stations were lower than the REF station, while not different from
the 1000E station. A comparison of the physiological results with the tissue
residues indicated slightly elevated PCB and PAH levels in the mussels col-
lected from the CNTR, 400E, and 1000E stations compared with the REF mussels.
154. These data may indicate exposure to BRH material during this de-
ployment period as one possible explanation for the SFG results. In order for
this to be true, however, some "resuspension event" would have had to occur at
the CLIS disposal site. The negative physiological response was not present
in the T + 15 mussels, collected 6 weeks earlier, and also absent in the
T + 27 mussels, which were placed in the field at T + 15 and collected on
T + 27. The cause of the observed response would have had to occur between
T + 15 and T + 21, but disappear after that because it was missing at T + 27.
155. Another example of an unexplained SFG effect was the T + 43 col-
lection. Mussels collected at the CNTR station exhibited a dramatic reduction
in clearance rate, absorption efficiency, and SFG compared with the other two
stations (the 1000E station was missing). These data indicate that the mus-
sels at this station were impacted by something. Resuspension of bottom sedi-
ments may once again provide a possible explanation. This deployment occurred
during the winter months when storms are typically stronger than during the
summer and fall. If bottom sediments containing BRH material were resuspended
and filtered out of the water by the mussels 1 m above the bottom, the clear-
ance rate and SFG results could be explained based on the results of labora-
tory experiments. However, corresponding tissue residue concentrations were
not elevated in the mussels from any station. In addition, not even labora-
tory exposures to 10 mg/£ of BRH suspended sediment affected absorption effi-
ciencies in this manner. The exact cause of the observed effects cannot be
explained based on BRH material alone; however, the magnitude of the response
would suggest that it is not a measurement artifact.
156. The final collection (T + 116), a 1-month deployment, indicated
that SFG values were not different among stations; however, they were all very
92
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low. Several plausible reasons are presented. First, temperatures In CLIS
were about 22° C, which may have stressed the animals. Secondly, there was an
abnormal occurrence at the Narragansett Bay reference site at the time mussels
were collected for deployment in CLIS. An incredible bloom of a small (<2-u)
algal species occurred throughout Narragansett Bay. This alga, present in
concentrations of greater than 1 billion cells/litre, caused the mussels and
some other bivalves in the Bay to cease feeding, resulting in mass mortalities
in mussel populations. This condition was noted 1 month prior to the T + 116
collection (June 1985) and persisted until the middle of August 1985. As a
result, mussels deployed at this time were not in the best physiological
condition.
157. To summarize the field experiments, exposure data generated inde-
pendently from water chemistry samples and tissue residues indicated that
maximum exposure to BRH material occurred during the disposal operation and
decreased rapidly thereafter. Of the range estimated for BRH concentrations
in the field, the lower estimate may be closer to the actual value. The maxi-
mum estimated exposure in the field, between 1.4 and 0.8 mg/£, was lower than
the lowest exposure concentration in the laboratory (1.5 mg/#.). Elevated tem-
peratures at T + 8 may have increased the sensitivity of mussels to BRH mate-
rial, which would explain the reduced SFG of mussels at the CNTR station
during this collection. Reduced residues after T + 8 were probably too low to
elicit a negative physiological impact on the mussels. Therefore, the effect
of BRH material on the SFG of mussels 1 m off the bottom was minimal. Mussels
deployed in CLIS longer than 1 month indicated that there were apparent SFG
differences on only two other occasions, T + 21 and T + 43, and both were
difficult to attribute to the BRH material alone. From the field data, a BRH
exposure concentration slightly greater than 1.4 to 0.8 mg/X, would be esti-
mated to cause an adverse effect on SFG in mussels. This level may decrease
as temperatures increase and mussels are less able to compensate for the BRH
material.
Laboratory-to-Field Comparison
158. The purpose of the laboratory-to-field comparison was to expose
mussels to BRH material in the laboratory and the field and compare the con-
centrations that produced a SFG effect in the mussels. The comparison of
93
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laboratory and field data indicated one obvious fact: laboratory and field
BRH exposures were different. The two independent estimates of BRH exposure
indicated that the maximum exposure in the field was lower than BRH exposures
in the laboratory. Cluster analysis of laboratory and field mussel residue
data yielded results similar to those obtained in estimating the field expo-
sures. That is, the mussel residues in the field were most similar to mussels
exposed to reference sediment in the laboratory. Considering these data, dis-
cussion of the laboratory-field comparison will focus initially on when condi-
tions (exposures and residues) were similar in the laboratory and field, and
secondly on estimated concentrations of BRH suspended material required to
affect SFG.
159. The exposure and residue data indicated that the most legitimate
comparison between laboratory and field SFG data was between all field samples
(with the exception of 400E at T + 2) and laboratory mussels exposed to
0-percent BRH. Furthermore, because temperature and season during the T = 0,
T + 2, and T + 55 collections were most similar to the laboratory exposures,
these SFG values should provide the most accurate laboratory-to-field compari-
son. The mean SFG value of mussels exposed to 0-percent BRH in the laboratory
for 28 days was 7.2 J/hr. The field-exposed mussels exhibited SFG values of
9.5, 8.9, and 6.5 J/hr, respectively, for the T = 0 (1000E), T = 0 (REF), and
T + 2 (1000E) collections. The similarity of these SFG values indicated that
the relative physiological conditions of these mussels were the same when en-
vironmental conditions were most alike. The mussel collection at T + 55 also
occurred in the spring at water temperatures similar to the laboratory expo-
sures; however, these mussels had been deployed in CLIS for 8 months. The SFG
values of these mussels, 3.0, 3.9, and 5.5 J/hr, were slightly lower than in
the 1-month laboratory exposure (7.7 J/hr). This may be attributable to the
difference in length of exposure.
160. The purpose of this qualitative comparison between laboratory and
field was to establish whether similar exposures produced similar results. In
light of the differences between the actual laboratory exposures (constant ex-
posure levels, food supply, etc.) and field exposures (fluctuating particulate
levels, food quantity, etc.), the SFG values between the two exposures were
relatively similar.
161. A second aspect of the laboratory-to-field comparison was a quali-
tative evaluation of the estimate of BRH material required to produce a SFG
94
-------
effect in the laboratory and field. The SFG, clearance rate, and actual
growth measurements during laboratory experiments provided a clear signal that
exposure to as little as 1.5 mg/£ of BRH material negatively affected
M. edulis. From these data, an estimated BRH exposure approximating 1.5 mg/I
should be sufficient to adversely affect mussels exposed in the field. While
the estimated maximum exposure concentration in the field (1.4-0.8 mg/£,
T + 2) did not appear to affect the mussels, an estimated lower concentration
(0.7-0.2 mg/Jl, CNTR station at T + 8) apparently did. This apparent contra-
diction was interpreted in terms of increased water temperatures during the
T + 8 collection and consequently greater sensitivity of the mussels to BRH
material. Because of the probable interactive effect of temperature, a quali-
tative estimate of the field BRH exposure concentration necessary to produce a
SFG effect would be between 0 and 2.0 mg/Jl.
162. Comparison of these two values indicated that the field estimate
of 0.7 to 0.2 mg/Jt was in the range predicted from laboratory experiments
(less than or equal to 1.5 mg/£). The possible existence of an effective
threshold BRH concentration between 0 and 1.5 mg/£, suggested by the labora-
tory dose-response curve, may help to explain the lack of effect in the field
and the presence of one in the laboratory at comparable temperatures. In
addition, the possibility of temperature-related sensitivity to this material
would help to explain the presence of an effect at lower exposure levels.
Nonetheless, the laboratory and field estimates provide a good qualitative
comparison of the effects of BRH material on mussels.
95
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PART V: CONCLUSIONS
163. The research described in this report evaluated the effects of a
dredged material on the physiological condition of M. edulis after laboratory
and field exposures. The results are as follows:
a. Laboratory dosing systems were successfully developed to
expose mussels to relatively constant concentrations of sus-
pended sediment.
b. A very good relationship was found between BRH laboratory
exposure level and tissue residue concentration for higher
molecular weight organic compounds such as PCBs. Lower molec-
ular weight PAHs, such as phenanthrene, were apparently metab-
olized and/or excreted by the mussels.
c. An inverse relationship was observed in the laboratory between
SFG and BRH exposure concentration. Likewise, SFG was in-
versely related to the tissue residue concentrations of some
of the contaminants present in the dredged material.
d. Lower SFG values in BRH-exposed mussels were attributable to
reduced clearance rates observed in the laboratory. In addi-
tion, mussels with lower SFG values exhibited reduced shell
growth rates.
e. The results of the laboratory study indicated that the mussel
residue concentrations were indicative of exposure conditions
and that the SFG index was useful for measuring the subsequent
biological effects of those exposures.
f. Independent estimates of field BRH concentrations, made from
~ tissue residues as well as from water chemistry data, indi-
cated that maximum exposure to mussels occurred during the
disposal operation and decreased rapidly (2 weeks) thereafter.
The maximum estimated concentration in the field (1.4 to
0.8 mg/X.) was lower than that in the lowest concentration used
in the laboratory experiments (1.5 mg/£).
g. The effect of the BRH exposures on the SFG of mussels, 1 m
above the bottom, was minimal. Reductions in SFG, attrib-
utable to BRH exposure, occurred only one time, 8 weeks
postdisposal. This effect was possibly due to increased sen-
sitivity of mussels to dredged material as a result of ele-
vated water temperatures.
h. The results of the field portion of this study indicated that
~ the SFG of mussels was not affected because the BRH exposure
concentrations were minimal.
i. A qualitative comparison between the SFG of mussels after lab-
~~ oratory and field exposures indicated that the estimated BRH
concentration which affected SFG in the field (0.2 mg/£.) was
similar to the range predicted from the laboratory experiments
(1.5 mg/£).
96
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Gonzalez, J. G., Everich, D., Hyland, J., and Melzian, B. D. 1979. "Effects
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APPENDIX A: CHEMICAL FORMULAS AND FIELD MUSSEL RESIDUE CONCENTRATIONS
-------
Table A1
Chemical Contaminants Selected for Measurement
in Both Field and Laboratory Studies
Chlorinated hydrocarbon pesticides
Polychlorinated biphenyls
Ethylan
Aromatic hydrocarbons > molecular weight 166
Compound Class Molecular Weight
Fluorene 166
C-l* Fluorene 180
C-2* Fluorene 194
C-3* Fluorene 208
C-4* Fluorene 222
Phenanthrene 178
Anthracene 178
C-l*Phenanthrene/anthracene 192
C-2*Phenanthrene/anthracene 206
C-3*Phenanthrene/anthracene 220
C-4*Phenanthrene/anthracene 234
Fluoranthene 202
Pyrene 202
C-l*Fluoranthene/pyrene 216
C-2*Fluoranthene/pyrene 230
C-3*Fluoranthene/pyrene 244
C-4*Fluoranthene/pyrene 258
Benzanthracene/chrysene** 228
C-l*Benzanthracene/chrysene** 242
C-2*Benzanthracene/chrysene** 256
C-3*Benzanthracene/chrysene** 270
C-4*Benzanthracene/chrysene** 284
Benzofluoranthenes 252
Benzo(e)pyrene 252
Benzo(a)pyrene 252
Perylene 252
(Continued)
* C-l, C-2, C-3, and C-4 refer to the number of methyl groups substituted
somewhere in the parent compound.
** These names are representative of the class of polynuclear aromatic
hydrocarbons (PAHs) measured at each molecular weight.
A3
-------
Table A1 (Concluded)
Compound Class Molecular Weight
C-l*Benzopyrene/perylene** 266
C-2*Benzopyrene/perylene** 280
C~3*Benzopyrene/perylene** 294
C-4*Benzopyrene/perylene** 308
Benzoperylene** 276
Dibenzanthracene** 278
Coronene 300
Dibenzocrysene** 302
Hetrocyclic aromatic compounds
Dibenzothiophene 184
C~l*dibenzothiophene 198
C-2*dibenzothiophene 212
C-3*dibenzothiophene 226
Metals
C-4*dibenzothiophene 240
Cadmium
Copper
Chromium
Iron
Lead
Manganese
Nickel
Zinc
* C-l, C-2, C-3, and C-4 refer to the number of methyl groups substituted
somewhere in the parent compound.
** These names are representative of the class of polynuclear aromatic
hydrocarbons (PAHs) measured at each molecular weight.
A4
-------
Table A2
Complete Formulae for Calculating all SUM and CENT Variable
PSUM
= POS166
+
POS178
+
POS202
+
POS228
+
POS252
+
POS276
+
POS278
+
POS300
+
POS302
HSUM
- H1C166
+
H2C166
+
H3C166
+
H4C166
+
H1C178
+
H2C178
+
H3C178
+
H4C178
+
H1C202
+
H2C202
+
H3C202
+
H4C202
+
H1C228
+
H2C228
+
H3C228
+
H4C228
+
H1C252
+
H2C252
+
H3C252
+
H4C252
SUM
= POS166
+
H1C166
+
H2C166
+
H3C166
+
H4C166
+
POS178
+
H1C178
+
H2C178
+
H3C178
+
H4C178
+
POS202
+
H1C202
+
H2C202
+
H3C202
+
H4C202
+
POS228
+
H1C228
+
H2C228
+
H3C228
+
H4C228
+
POS252
+
H1C252
+
H2C252
+
H3C252
+
H4C252
+
POS276
+
POS278
+
POS300
+
PCENT =
HCENT *=
P0S302
fPOS166*166
POS276*276
+ POS178*178 + P0S202*202
+ POS278*278 + POS300*300
[H1C166*180 + H2C166*194
H2C178*206 + H3C178*220
H3C202*244 + H4C202*258
H4C228*284 + H1C252*266
HSUM
+ H3C166*208
+ H4C178*234
+ H1C228*242
+ H2C252*280
+ POS228*228 + POS252*252 +
+ POS302*302]/PSUM
+ H4C166*222 + H1C178*192 +
+ H1C202*216 + H2C202*230 +
+ H2C228*256 + H3C228*270 +
+ H3C252*294 + H4C252*308]/
CENT =
[POS166*166
POS178*178
POS202*202
POS228*228
POS252*252
POS276*276
H1C166*180
H1C178*192
H1C202*216
H1C228*242
H1C252*266
POS278*278
H2C166*194
H2C178*206
H2C202*230
H2C228*256
H2C252*280
P0S300*300
+ H3C166*208
+ H3C178*220
+ H3C202*244
+ H3C228*270
+ H3C252*294
+ H4C166*222 +
+ H4C178*234 +
+ H4C202*258 +
+ H4C228*284 +
+ H4C252*308 +
+ POS302*302]/SUM
The sum of alkyl homologs of PAH molecular weight 178 (HOS178) is calculated
according to the following formula:
HOS178 - H1C178 + H2C178 + H3C178 + H4C178
where
HOS178 = sum of C-l to C-4 alkyl-substituted 178 PAHs
This statistic was chosen to describe the alkyl homologs because the 178 alkyl
homologs are the most intense homologs within the Black Rock Harbor (BRH) PAH
distribution and because they afford the greatest BRH to REFS concentration
ratio. Alkyl homologs were included because of potential differences between
them and parent PAHs.
A5
-------
Table A3
Tissue Residue Concentrations in Mussels from the
T - 4 Field Collection in CLIS (22 Apr 83)*
Station
Chemical Compound CNTR 400E 1000E REFS
Phenanthrene 210.0 117.0 98.0 38.0
Sum of 178 alkyl 580.0 310.0 310.0 290.0
homologs
Fluoranthene 161.0 102.0 90.0 82.0
Benzo(a)pyrene 37.0 20.0 34.0 25.0
Ethylan 5.0 3.0 5.0 10.0
PCB as A1254 380.0 270.0 400.0 440.0
SUM of PAHs 2,600.0 1,520.0 1,650.0 1,380.0
CENTROID of PAHs 218.0 219.0 225.0 228.0
Copper 13.5 15.1 14.5 12.5
Cadmium 1.9 1.8 1.8 1.8
Chromium 1.8 3.8 2.2 1.6
Iron 370.0 1,400.0 530.0 340.0
* Units are nanograms per gram dry weight for the organic compounds and the
statistic SUM, micrograms per gram dry weight for the inorganic elements,
and molecular weight for the statistic CENTROID.
A6
-------
Table A4
Tissue Residue Concentrations in Mussels from the T + 0 Field Collection in
CLIS (24 May 83).* The CNTR Station Was Not Deployed Because of
the Dumping Operation, and the 400F. Station Was Lost
Station
Chemical Compound
Phenanthrene
Sum of 178 alkyl
homologs
Fluoranthene
Benzo(a)pyrene
Ethylan
PCB as A1254
SUM of PAHs
CENTROID of PAHs
Copper
Cadmium
Chromium
Iron
1000E
43.0
1,440.0
161.0
100.0
102.0
1,080.0
5,400.0
230.0
16.5
2.0
2.6
420.0
REFS
16.0
290.0
52.0
18.0
9.0
500.0
1,290.0
232.0
10.9
2.3
1.5
330.0
* Units are nanograms per gram dry weight for the organic compounds and the
statistic SUM, micrograms per gram dry weight for the inorganic elements,
and molecular weight for the statistic CENTROID.
A7
-------
Table A5
Tissue Residue Concentrations in Mussels from the T + 2 Field Collection in
CLIS (07 June 83).* The CNTR Station Was Not Deployed Because
of the Disposal Operation
Station
Chemical Compound
Phenanthrene
Sum of 178 alkyl
homologs
Fluoranthene
Benzo(a)pyrene
Ethylan
PCB as A1254
SUM of PAHs
CENTROID of PAHs
Copper
Cadmium
Chromium
Iron
400E
69.0
1,900.0
290.0
210.0
71.0
1,440.0
8,700.0
232.0
16.9
2.3
3.0
510.0
1000E
41.0
970.0
126.0
118.0
39.0
1,020.0
4,700.0
234.0
15.6
2.3
3.0
560.0
REFS
13.0
540.0
72.0
51.0
17.0
630.0
2,500.0
233.0
10.8
1.9
2.0
560.0
* Units are nanograms per gram dry weight for the organic compounds and the
statistic SUM, micrograms per gram dry weight for the inorganic elements,
and molecular weight for the statistic CENTROID.
A8
-------
Table A6
Tissue Residue Concentrations in Mussels from the
T + 8 Field Collection in CLIS (10 Jul 83)*
Station
Chemical Compound CNTR 400E 1000E REFS
Phenanthrene 11.0 14.0 9.0 7.0
Sum of 178 alkyl 350.0 340.0 193.0 105.0
homologs
Fluoranthene 45.0 46.0 31.0 23.0
Benzo(a)pyrene 40.0 50.0 18.0 20.0
Ethylan 22.0 20.0 7.0 1.0
PCB as A1254 700.0 740.0 620.0 480.0
SUM of PAHs 1,870.0 2,100.0 1,020.0 760.0
CENTROID of PAHs 234.0 236.0 231.0 240.0
Copper 10.1 9.6 11.5 4.4
Cadmium 1.9 2.0 1.3 0.9
Chromium 1.4 1.4 3.2 0.8
Iron 340.0 370.0 820.0 240.0
* Units are nanograms per gram dry weight for the organic compounds and the
statistic SUM, micrograms per gram dry weight for the inorganic elements,
and molecular weight for the statistic CENTROID.
A9
-------
Table A7
Tissue Residue Concentrations in Mussels from the
T -f 12 Field Collection in CLIS (10 Aug 83)*
Station
Chemical Compound
Phenanthrene
Sum of 178 alkyl
homologs
Fluoranthene
Benzo(a)pyrene
Ethylan
PCB as A1254
SUM of PAHs
CENTROID of PAHs
Copper
Cadmium
Chromium
Iron
CNTR
17.0
250.0
41.0
41.0
9.0
640.0
1,600.0
237 .0
5.3
0.9
1.0
164.0
400E
10.0
160.0
28.0
17.0
8.0
660.0
940.0
236.0
5.6
0.9
0.7
167.0
1000E
9.0
96.0
20.0
16.0
3.0
550.0
710.0
239.0
7.5
1.2
1.6
450.0
REFS
8.0
65.0
15.0
13.0
1.0
570.0
530.0
240.0
5.8
1.1
0.7
177.0
* Units are nanograms per gram dry weight for the organic compounds and the
statistic SUM, micrograms per gram dry weight for the inorganic elements,
and molecular weight for the statistic CENTROID.
A10
-------
Table A8
Tissue Residue Concentrations in Mussels from the
T + 15 Field Collection in CLTS (06 Sep 83)*
Station
Chemical Compound CNTR 400E 1000E REFS
Phenanthrene 13.0 9.0 10.0 6.0
Sum of 178 alkyl 370.0 230.0 210.0 43.0
homologs
Fluoranthene 57.0 38.0 33.0 14.0
Benzo(a)pyrene 53.0 45.0 28.0 7.0
Ethylan 10.0 6.0 4.0 1.0
PCB as A1254 870.0 630.0 640.0 550.0
SUM of PAHs 2,100.0 1,540.0 1,240.0 350.0
CENTROID of PAHs 236.0 239.0 237.0 238.0
Copper 7.7 6.0 8.0 5.8
Cadmium 1.0 1.1 1.1 0.9
Chromium 1.2 0.9 1.1 0.9
Iron 260.0 179.0 290.0 260.0
* Units are nanograms per gram dry weight for the organic compounds and the
statistic SUM, micrograms per gram dry weight for the inorganic elements,
and molecular weight for the statistic CENTROID.
All
-------
Table A9
Tissue Residue Concentrations in Mussels from the
T + 21 Field Collection in CLIS (18 Oct 83)*
Chemical Compound
Phenanthrene
Sum of 178 alkyl
homologs
Fluoranthene
Benzo(a)pyrene
Ethylan
PCB as A1254
SUM of PAHs
CENTROID of PAHs
Copper
Cadmium
Chromium
Iron
Station
CNTR 400E 1000E REFS
12.0 11.0 11.0 10.0
132.0 101.0 88.0 46.0
33.0 25.0 22.0 16.0
24.0 9.0 17.0 9.0
2.0 2.0 1.0 0.0
540.0 680.0 570.0 420.0
1,000.0 670.0 700.0 400.0
240.0 234.0 239.0 238.0
22.1 16.3 15.1 16.4
5.1 4.4 4.8 5.0
2.3 2.6 2.2 2.2
440.0 540.0 420.0 480.0
* Units are nanograms per gram dry weight for the organic compounds and the
statistic SUM, micrograms per gram dry weight for the inorganic elements,
and molecular weight for the statistic CENTROID.
A12
-------
Table A10
Tissue Residue Concentrations in Mussels from the T + 27 Field Collection in
CLIS (29 Nov 83).* The CNTR Station Was Missing
at the Time of Collection
Chemical Compound
Phenanthrene
Sum of 178 alkyl
homologs
Fluoranthene
Benzo(a)pyrene
Ethylan
PCB as A1254
SUM of PAHs
CENTROID of PAHs
Copper
Cadmium
Chromium
Iron
Station
400E
18.0
230.0
68.0
39.0
3.0
540.0
1,820.0
240.0
16.4
3.2
2.5
570.0
1000E
10.0
117.0
36.0
32.0
1.0
380.0
1,150.0
244.0
21.0
3.4
3.6
920.0
REFS
8.0
86.0
37.0
19.0
0.0
450.0
860.0
240.0
23.3
3.6
3.3
920.0
* Units are nanograms per gram dry weight for the organic compounds and the
statistic SUM, micrograms per gram dry weight for the inorganic elements,
and molecular weight for the statistic CENTROID.
A13
-------
Table All
Tissue Residue Concentrations in Mussels from the T + 43 Field Collection in
CLIS (20 Mar 84).* Metals Were Not Measured for These Samples.
The 1000E Station Was Missing at the Time of Collection
Station
Chemical Compound CNTR 400E REFS
Phenanthrene 16 18 17
Sum of 178 alkyl 94 78 70
homologs
Fluoranthene 28 26 24
Benzo(a)pyrene 8 4 7
Ethylan 2 1 1
PCB as A1254 350 330 280
SUM of PAHs 510 460 450
CENTROID of PAHs 229 230 231
* Units are nanograms per gram dry weight for the organic compounds and the
statistic SUM, and molecular weight for the statistic CENTROID.
A14
-------
Table A12
Tissue Residue Concentrations in Mussels from the T + 55 Field Collection in
CLIS (05 June 84).* Metals Were Not Measured in These Samples.
The CNTR Station Was Missing at the Time of Collection
Station
Chemical Compound 400E 1000E REFS
Phenanthrene 6 6 4
Sum of 178 alkyl 89 91 54
homologs
Fluoranthene 25 31 18
Benzo(a)pvrene 6 7 6
Ethylan 0 1 0
PCB as A1254 540 490 550
SUM of PAHs 520 550 370
CENTROID of PAHs 234 235 236
* Units are nanograms per gram dry weight for the organic compounds and the
statistic SUM, and molecular weight for the statistic CENTROID.
A15
-------
Table A13
Tissue Residue Concentrations in Mussels from the
T + 116 Field Collection in CLIS (13 Aug 85)*
Chemical Compound CNTR
Phenanthrene 3.'
Sum of 178 alkyl 79.
homologs
Fluoranthene 18.
Benzo(a)pyrene 18.
Ethylan 1.
PCB as A1254 310.0
SUM of PAHs 700.0
CENTR0ID of PAHs 242.0
Copper 8.5
Cadmium 1 • 5
Chromium 1.2
Iron 290.0
Station
400E 1000E REFS
6.0 3.0 3.0
124.0 80.0 58.0
27.0 24.0 19.0
40.0 22.0 17.0
2.0 1.0 0.0
350.0 450.0 440.0
1,270.0 810.0 620.0
243.0 241.0 244.0
7.5 6.9 7.6
1.3 1.3 1.3
1.1 0.9 0.9
260.0 220.0 220.0
Units are nanograms per gram dry weight for the organic compounds and the
statistic SUM, micrograms per gram dry weight for the inorganic elements
and molecular weight for the statistic CENTROID.
A16
-------
APPENDIX B: COMPARISON OF SELECTED CONTAMINANTS IN BRH AND REF SEDIMENTS
-------
The contaminant-specific analysis of the BRH and REF sediments is
presented in summary form for the representative subset of chemical compounds
discussed in this report. These analyses demonstrate clearly the differences
in contaminant concentration between the two sediments.
Table B1
Concentrations of the Ten Selected Contaminants and Two Summary Statistics
for Both BRH and REF Sediments. Means ± Standard Deviations
Sediment*
Chemical Compound BRH REF
Phenanthrene 5,200 ± 1,820 (8)** 85 ± 16 (12)
Sum of 178 alkyl 29,000 ± 8,400 (8) 100 ± 19 (12)
homologs
Fluoranthene 6,500 ± 1,400 (8) 240 ± 31 (12)
Benzo(a)pyrene 4,000 ± 950 (8) 251 ± 27 (12)
Ethylan 3,900 ± 750 (8) 0 ± -- (12)
PCB as A1254 7,000 ± 1,560 (8) 39 ± 4 (12)
SUM of PAHs 146,000 ± 31,000 (8) 4,500 ± 490 (12)
CENTR0ID of PAHs 232.7 ± 1.6 (8) 249.2 ± 1.6 (12)
Copper 2,900 ± 310 (54) 60 ± 3 (45)
Cadmium 24 ± 1.0 (54) 0.23 ± 0.04 (45)
Chromium 1,480 ± 104 (54) 50 ± 15 (45)
Iron 31,000 ± 2,800 (54) 21,000 ± 1,400 (45)
* Units are nanograms per gram dry weight for the organic compounds and the
statistic SUM, micrograms per gram dry weight for the inorganic elements,
and molecular weight for the statistic CENTROID.
** (N) = number of replicates.
B3
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