FIELD VERIFICATION PROGRAM
(AQUATIC DISPOSAL)
TECHNICAL REPORT D-88-4
A FIELD AND LABORATORY STUDY USING
ADENYLATE ENERGY CHARGE AS AN INDICATOR
OF STRESS IN MYTILUS EDULIS AND NEPHTYS
INCISA TREATED WITH DREDGED MATERIAL
Gerald E. Zaroogian, Peter F Rogerson
Gerald Hoffman, Mary Johnson
Environmental Research Laboratory
US Environmental Protection Agency
Narragansett, Rhode Island 02882
D. Michael Johns
Tetra Tech
Bellevue, Washington 98005
Science Applications International Corporation
Narragansett, Rhode Island 02882
Approved For Public Release; Distribution Unlimited
Prepared for DEPARTMENT OF THE ARMY
US Army Corps of Engineers
Washington, DC 20314-1000
US Environmental Protection Agency
Washington, DC 20460
Monitored by Environmental Laboratory
US Army Engineer Waterways Experiment Station
PO Box 631, Vicksburg, Mississippi 39180-0631
by
and
William G. Nelson
i/im
June 1988
Final Report
and

-------
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)

-------
SUBJECT: Transmittal of Field Verification Program Technical Report Entitled
"A Field and Laboratory Study Using Adenylate Energy Charge as an
Indicator of Stress in Mytilus edulis and Nephtys incisa Treated
with Dredged Material"
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.

-------
SUBJECT: Transmittal of Field Verification Program Technical Report Entitled
"A Field and Laboratory Study Using Adenylate Energy Charge as an
Indicator of Stress in Mytilue edulis and Nephtys inaisa Treated
with Dredged Material"
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.
Smash Choromokos, Jr., Hi.
Choromokos, Jr., Pfi.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

-------
Unclassi fieri
SECURITY CLASSIFICATION OF THIS PAGE
REPORT DOCUMENTATION PAGE
form Approved
OMB No 0704-0188
Exp Date Jun 30, 1986
1a REPORT SECURITY CLASSIFICATION
Unclassified
b 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-88-4
6a NAME OF PERFORMING ORGANIZATION
See reverse
6b OFFICE SYMBOL
(If applicable)
7a NAME OF MONITORING ORGANIZATION
USAEWES
Environmental Laboratory
6c. ADDRESS (City, State, and ZIP Code)
Narragansett, RI 02882;
Bellevue, WA 98005;
Narragansett, RI 02882
7b ADDRESS (City, State, and ZIP Code)
P0 Box 631
Vicksburg, MS 39180-0631
8a. NAME OF FUNDING /SPONSORING
ORGANIZATION
See reverse
8b OFFICE SYMBOL
(If applicable)
9 PROCUREMENT INSTRUMENT IDENTIFICATION NUMBER
8c. ADDRESS (City, State, and ZIP Code)
Washington, DC 20314-1000;
Washington, DC 20460
10. SOURCE OF FUNDING NUMBERS
PROGRAM
ELEMENT NO
PROJECT
NO
TASK
NO
WORK UNIT
ACCESSION NO
.TITLE (Include Security Classification)
A Field and Laboratory Study Using Adenylate Energy Charge as an Indicator of Stress in
Mytilue edulis and Nephtys -inoiea Treated with Dredged Material
12 PERSONAL AUTHOR(S) Zaroogian, Gerald E; Rogerson, Peter F.; Hoffman, Gerald;
Johnson. Marvi Johns._D. Michael: Nelson. Williams G
13a TYPE OF REPORT
Final report
13b TIME COVERED
FROM	TO
14 DATE OF REPORT (Year, Month, Day)
May 1988
15 PAGE COUNT
166
16 SUPPLEMENTARY NOTATION
Available from National Technical Information Service, 5285 Port Royal Road, Springfield,
VA 22161.
17 COSATI CODES
18 SUBJECT TERMS (Continue on reverse if necessary and identify by block number)
FIELD
GROUP
SUB-GROUP






19. ABSTRACT (Continue on reverse if necessary and identify by block number)
A study was conducted to test the applicability of adenylate energy charge (AEC) and
adenine nucleotide pool concentrations as measures of biological response in the blue mus-
sel, Mytilue edulie, and the marine polychaete, Nephtys ineisa, after exposure in the lab-
oratory and field to contaminated dredged material from Black Rock Harbor (BRH), Bridge-
port, Conn. A second objective was to include field verification of laboratory results,
and a third objective was to investigate residue-effect relationships between tissue con-
centrations of BRH contaminants and AEC and adenine nucleotide pool concentrations. This
project was part of the US Environmental Protection Agency/Corps of Engineers Field
Verification Program.
Biological responses were measured in a laboratory dosing system that provided con-
stant exposure concentrations of suspended BRH sediment ranging from 0 to 10 mg/t for
(Continued)
20 DISTRIBUTION/AVAILABILITY OF ABSTRACT
H UNCLASSIFIED/UNLIMITED ~ SAME AS RPT
~ DTIC USERS
21 ABSTRACT SECURITY CLASSIFICATION
Unclassified
22a. NAME OF RESPONSIBLE INDIVIDUAL
22b TELEPHONE (Include Area Code) 22c OFFICE SYMBOL
DO FORM 1473,84 mar
83 APR edition may be used until exhausted
All other editions are obsolete
SECURITY CLASSIFICATION Of THIS PAGE
Unclassified

-------
Unclassified
•CCUMITY ClAMIFICATION THI* *Aซซ
6a. NAME OF PERFORMING ORGANIZATION (Continued).
US Environmental Protection Agency;
Tetra Tech;
Science Applications International Corporation
8a. NAME OF FUNDING/SPONSORING ORGANIZATION (Continued).
DEPARTMENT OF THE ARMY
US Army Corps of Engineers;
US Environmental Protection Agency
19. ABSTRACT (Continued).
M. edulis and 0 to 200 mg/H for N. inoisa. In the field, biological responses were mea-
sured in both species sampled along a transect of stations at the Central Long Island
Sound disposal site. Strong exposure-residue relationships measured in laboratory exper-
iments indicated that selected contaminants in BRH sediments were biologically available.
Tissue residue concentrations, particularly of persistent compounds such as polychlori-
nated biphenyls, were found to be closely related to exposure concentration. This close
relationship between exposure concentrations and tissue residues, as defined in laboratory
experiments, was used to estimate field exposures based on tissue residues measured in
field-collected M. edulis and N. inoisa. The field exposures estimated from tissue resi-
dues were corroborated using estimates based on water and sediment chemistry.
The biological responses evaluated in this report included the adenine nucleotide
measures of adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, ade-
nylate pool, and AEC. These responses were measured in M. edulis and N. inoisa exposed to
BRH sediment in the laboratory and the field. The only significant laboratory response
was a reduction in adenylate pool concentration measured in M. edulis at BRH exposure con-
centrations higher than any estimated exposures in the field. The only significant field
responses were station-related changes in all adenylate nucleotide concentrations measured
in N. inoisa 16 weeks postdisposal and were indicative of nonstressed organisms. This
represented an exposure lasting 10 weeks longer than the longest laboratory exposure,
which was 6 weeks.
The adenine nucleotide pool concentrations in organisms exposed to BRH sediments
will respond in a concentration-related manner. However, these responses are relatively
insensitive in M. edulie and are related to long exposure periods in N. inoisa.
The apparent differences between laboratory and field responses for M. edulis and
N. inoisa could be explained by differences in exposures between the two situations. For
M. edulis, the exposures were much higher in the laboratory. For N. inoisa, the exposures
in the laboratory and the field were comparable, but the field response was significant at
16 weeks postdisposal. This length of exposure far exceeded the length of any laboratory
experiments.
Adenine nucleotides and AEC are important in energy transformation and in regulation
of metabolic processes. Therefore, it is not surprising that responses in adenine nucleo-
tide pools correlate with tissue concentrations of BRH contaminants in exposed organisms.
Measurement of the adenine nucleotide concentrations may help to characterize the energy
costs incurred by organisms under stressful conditions.
UnclaBB^pH
HCUNITY CLAUDICATION Or THII PAOI

-------
PREFACE
This report describes work performed by the US Environmental Protection
Agency (USEPA), Environmental Research Laboratory, Narragansett, R. I. (ERLN),
as part of the Interagency Field Verification of Testing and Predictive Meth-
odologies for Dredged Material Disposal Alternatives Program (Field Verifica-
tion Program (FVP)). The FVP was sponsored by the Office, Chief of Engineers
(OCE), US Army, 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 option conducted by WES.
The principal investigators for this aquatic study and authors of this
report were Drs. Gerald E. Zaroogian, ERLN; D. Michael Johns, Tetra Tech;
Peter F. Rogerson and Gerald Hoffman, ERLN; and Mr. William G. Nelson, Science
Applications International Corporation (SAIC). Laboratory-cultured algae were
provided by Mr. Greg Tracey, SAIC. Technical support for the adenylate energy
charge measurements was provided by Ms. Mary Johnson, ERLN. Diving support
for the field portion of the study was provided by Messrs. Bruce Reynolds and
Norman Rubinstein, ERLN, and Greg Tracey, SAIC.
Analytical chemistry support was provided by 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 S. Gardiner, and Colette J. Brown, Com-
puter Science Corporation (CSC), provided word processing support in the prep-
aration of this report. Predictive models for field exposures were supplied
by Drs. John F. Paul, ERLN, and Wayne R. Munns, SAIC. In addition, assistance
in statistical analysis was provided by Dr. James Heltshe and Mr. Jeffery
Rosen, CSC. Critical reviews of this report were completed by Drs. Eugene
Jackim, John H. Gentile, and Gerald G. Peach, ERLN. Technical reviews were
provided by WES personnel.
The USEPA Technical Director for the FVP was Dr. Gentile; the Technical
Coordinators were Dr. Pesch and Mr. Walter Galloway. The OCE Technical
1

-------
Monitors were Drs. John Hall, Robert J. Pierce, and William L. Klesch.
The study was conducted under the direct WES management of
Drs. Thomas M. Dillon and Richard K. Peddicord and under the general manage-
ment 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 FVP Coordinator was
Mr. Robert L. Lazor, and the Environmental Effects of Dredging Program (EEDP)
Managers were Mr. Charles C. Calhoun, Jr., and Dr. Robert M. Engler.
Dr. Thomas D. Wright was the WES Technical Coordinator for the FVP reports.
This report was edited by Ms. Jamie W. Leach of the WES Information Technology
Laboratory.
COL Dwayne G. Lee, CE, was Commander and Director of WES. Dr. Robert W.
Whalln was Technical Director.
This report should be cited as follows:
Zaroogian, G, E., Rogerson, P. F., Hoffman, G., Johnson, M.,
Johns, D. M., and Nelson, W. G. 1988. "A Field and Laboratory
Study Using Adenylate Energy Charge as an Indicator of Stress in
Mytilus edulis and Nephtys incisa Treated with Dredged Material,"
Technical Report D-88-4, prepared by the US Environmental Pro-
tection Agency, Narragansett, R. I., for the US Army Engineer
Waterways Experiment Station, Vicksburg, Miss.
2

-------
CONTENTS
Page
PREFACE		1
LIST OF TABLES		4
LIST OF FIGURES		5
PART I: INTRODUCTION		8
Background		8
Project Description		10
Project Scope		12
Laboratory-to-Field Comparisons		13
Residue-Effects Relationships		13
Adenylate Energy Charge		13
PART II: MATERIALS AND METHODS		15
Laboratory Methods		15
Field Methods		29
Chemical Methods		34
Statistical Analysis Methods		38
PART III: RESULTS		40
Laboratory		40
Field		62
Laboratory-to-Field Comparisons		97
Residue-Effects Comparisons		104
PART IV: DISCUSSION		Ill
Laboratory Experiments		Ill
Field Experiments		114
Laboratory-to-Field Comparisons		117
Residue-Effects Comparisons		121
PART V: CONCLUSIONS		127
REFERENCES		129
APPENDIX A: BLACK ROCK HARBOR SEDIMENT PERCENTAGE CALCULATIONS		A1
APPENDIX B: CHEMICAL FORMULAS AND FIELD MUSSEL RESIDUE
CONCENTRATIONS		B1
3

-------
LIST OF TABLES
No.	Page
1	Collection Information for the M. edulis Used in the
Laboratory Experiments		15
2	Collection Information for the N. incisa Used in the
Laboratory Experiments		16
3	Cruise Number, Deployment Date, Retrieval Date, and Length
of Deployment for Mussels Transplanted to CLIS		30
4	Suspended Sediment Concentrations in the Mussel
Exposure System		40
5	Chemical Monitoring of the Exposure System in Experiment 2		42
6	Measured TSS Concentrations and Exposure Conditions for
Laboratory Tests with N. incisa		43
7	Chemical Analysis of Seawater in Exposure Chambers of 42-Day
Experiment Exposing N. incisa to BRH sediment		44
8	Concentrations of the Ten Selected Contaminants and Two
Summary Statistics for Both BRH and REF Sediments		45
9	PCB Tissue Residues in Mussels from Laboratory Experiment 1....	46
10	PCB Tissue Residues in Mussels from Laboratory Experiment 2....	46
11	PCB Concentrations in Mussels from Both Laboratory
Experiments		47
12	Adenine Nucleotide Concentrations in M. edulis from the Two
Laboratory Experiments		62
13	Adenine Nucleotide Concentrations in N. incisa from the Three
Laboratory Experiments			64
14	Predicted BRH Suspended Material Sediment Exposure Required
To Achieve the Measured Tissue Residue Values of Mussels
Deployed in CLIS		66
15	Predicted BRH Suspended Sediment Exposure Based on PCB and
Copper Whole Water Chemistry Data		69
16	Estimated Concentrations of BRH Sediment Suspended at
Sediment-Water Interface Based on PCB Concentrations
in Field-Collected N. incisa		70
17	Percent BRH Sediment in the Surficial Sediments at the
FVP Disposal Site		73
18	Concentration of BRH at the Sediment-Water Interface for
TSS Concentrations of 30 mg/JI and 10 mg/fl, and an
Enrichment of lOx		73
19	Adenine Nucleotide Concentrations Measured in Adductor
Muscle Tissues of M. edulis Sampled at FVP Field
Stations on Specific Dates		89
20	Adenine Nucleotide Concentrations Measured in N. incisa
Sampled on Specified Dates at the FVP Field Stations		95
21	Summary of P Values Indicating Degree of Statistical
Significance for Each Regression Analysis Between
Biological Variables and Tissue Concentrations of
Contaminants for Laboratory Samples of M. edulis.....	 105
22	Summary of P Values Indicating Degree of Statistical
Significance for Each Regression Analysis Between
Biological Variables and Tissue Concentrations for
Field Samples of M. edulis	 106
4

-------
LIST OF TABLES (Continued)
No.	Page
23	Summary of P Values Indicating Degree of Statistical
Significance for Each Regression Analysis Between
Biological Variables and Tissue Concentrations of
Contaminants for Laboratory Samples of N. inoisa		107
24	Summary of P Values Indicating Degree of Statistical
Significance for Each Regression Analysis Between
Biological Variables and Tissue Concentrations for
Field Samples of N. inoisa	 108
25	Summary of the Number of Significant Correlations Between
Adenine Nucleotide Concentrations and Tissue Contaminant
Concentrations by Biological Variable, by Species, and
by Laboratory Versus Field Categories	 109
26	Summary of the Number of Significant Correlations Between
Adenine Nucleotide Concentrations and Tissue Contaminant
Concentrations by Chemical Class, by Species, and
by Laboratory Versus Field Categories		110
LIST OF FIGURES
No.	Page
1	Central Long Island Sound disposal site and Black Rock
Harbor dredge site		10
2	FVP sampling stations		11
3	Suspended sediment dosing system		16
4	Suspended sediment oxidation system		18
5	Laboratory exposure system for M. edulis		19
6	Proportional diluter used to deliver suspended sediment
to the N. inoisa exposure chambers...		21
7	Nephtys inoisa exposure chamber		22
8	Summary of procedure for the extraction of adenine
nucleotides from the adductor muscle of M. edulis		24
9	Summary of procedure for the extraction of adenine
nucleotides from the tissue of N. inoisa		25
10	Summary of procedures for analysis of ATP, ADP, and
AMP in adductor muscle tissue of M. edulis and whole
N. inoisa		27
11	Concentrations of PCB as A1254 in the tissue of M. edulis
exposed to BRH suspended sediments for 28 days		47
12	Concentrations of PCB as A1254, normalized for lipids,
in the tissue of M. edulis exposed to BRH sediment
for 14 days		48
13	Concentrations of phenanthrene and sum of 178 alkyl homologs
in the tissue of M. edulis exposed to BRH suspended
sediments for 28 days		49
14	Concentrations of fluoranthene and benzo(a)pyrene in the
tissue of M. edulis exposed to BRH suspended sediments
for 28 days		50
5

-------
LIST OF FIGURES (Continued)
No.	Page
15	Concentrations of SUM of PAHs and CENT of PAHs in the
tissue of M, edulis exposed to BRH suspended sediments
for 28 days						51
16	Concentrations of ethylan and PCB as A1254 in the tissue
of M, edulis exposed to BRH suspended sediments
for 28 days		52
17	Concentrations of cadmium and copper in the tissue of
M. edulis exposed to BRH suspended sediments for 28 days		53
18	Concentrations of chromium and iron in the tissue of
M. edulis exposed to BRH suspended sediments for 28 days		54
19	Concentrations of phenanthrene and sum of 178 alkyl homologs
in the tissue of N. incisa exposed to BRH suspended
sediments for 42 days		56
20	Concentrations of fluoranthene and benzo(a)pyrene in the
tissue of N. incisa exposed to BRH suspended sediments
for 42 days		57
21	Concentrations of SUM of PAHs and CENT of PAHS in the
tissue of N. incisa exposed to BRH suspended sediments
for 42 days				58
22	Concentrations of ethylan and PCB as A1254 in the tissue of
N. incisa exposed to BRH suspended sediments for 42 days		59
23	Concentrations of cadmium and copper in the tissue of
N. incisa exposed to BRH suspended sediments for 42 days		60
24	Concentrations of chromium and iron in the tissue of
N. incisa exposed to BRH suspended sediments for 42 days		61
25	Response of adenine nucleotide pools in M. edulis to BRH
exposure in laboratory experiments		63
26	Response of adenine nucleotide pools in N. incisa to BRH
exposure in laboratory experiments			65
27	Suspended sediment concentrations from 1 m above the bottom
to the sediment-water interface for storm and background
conditions		72
28	Concentrations of phenanthrene and the sum of 178 alkyl
homologs in the tissues of M. edulis exposed at the
specified FVP stations and sampling dates		75
29	Concentrations of fluoranthene and benzo(a)pyrene in the
tissues of M. edulis exposed at the specified FVP
stations and sampling dates		76
30	Concentrations of the SUM of PAHs and CENT of PAHs in the
tissues of M. edulis exposed at the specified FVP stations
and sampling dates		77
31	Concentrations of PCB as A1254 and ethylan in the tissues of
M. edulis exposed at the specified FVP stations and
sampling dates		78
32	Concentrations of cadmium and copper in the tissues of
M. edulis exposed at the specified FVP stations and
sampling dates		79
33	Concentrations of chromium and iron in the tissues of
M. edulis exposed at the specified FVP stations and
sampling dates		80
6

-------
LIST OF FIGURES (Continued)
No.	Page
34	Concentrations of phenanthrene and the sum of 178 alkyl
homologs in the tissues of N. inaisa collected at the
specified FVP stations and sampling dates		82
35	Concentrations of fluoranthene and benzo(a)pyrene in the
tissues of N. inaisa collected at the specified FVP
stations and sampling dates		83
36	Concentrations of the SUM of PAHs and CENT of PAHs in the
tissues of N. inaisa collected at the specified FVP stations
and sampling dates		84
37	Concentrations of PCB as A1254 and ethylan in the tissues of
N. inaisa collected at the specified FVP stations and
sampling dates		85
38	Concentrations of cadmium and copper in the tissues of
N. inaisa collected at the specified FVP stations and
sampling dates							86
39	Concentrations of chromium and iron in the tissues of
N. inaisa collected at the specified FVP stations and
sampling dates				87
40	Relationship between adenine nucleotide concentration and
AEC in M. edulis and PCB tissue residue concentrations
in laboratory- and field-exposed animals		91
41	Relationship between total adenine nucleotide concentration
and AEC in N. inaisa and PCB tissue residue concentrations
in laboratory- and field-exposed animals		98
7

-------
A FIELD AND LABORATORY STUDY USING ADENYLATE ENERGY CHARGE
AS AN INDICATOR OF STRESS IN MYTILUS EPULIS AND
NEPHTYS INCTSA TREATED WITH DREDGED MATERIAL
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 contami-
nant bioaccumulation; and (d) describing the initial mixing during disposal.
These methods have been used for determining the suitability of dredged mate-
rial for open-water disposal. The procedures in this manual represented the
technical state of the art at that time and were never intended to be inflex-
ible methodologies. The recommended test methods were chosen to provide tech-
nical information 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).
3.	To meet this critical need, the Interagency Field Verification of
8

-------
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,
US Army, 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 test methodologies for predicting the environmental con-
sequences of dredged material disposal under aquatic, intertidal, and upland
conditions. The aquatic portion of the FVP was conducted by the USEPA En-
vironmental Research Laboratory, Narragansett, R. I. (ERLN). The intertidal
and upland portions, conducted by WES, are reported in separate documentation.
4.	The ERLN was responsible for conducting research on the aquatic
option for disposal of dredged material. There were three research objectives
for this portion of the program. The first was to demonstrate the applica-
bility of existing test methods for detecting and measuring the effects of
dredged material and to determine the degree of variability and reproduci-
bility inherent in the testing procedure. This phase of the program (Labora-
tory Documentation) is complete, and the results have been published in a
series of technical reports. This information provides insight into how the
various methods function, their sources of variability, their respective and
relative sensitivities to the specific dredged material being tested, and the
degree of confidence that can be placed on the data derived from the applica-
tion of the methods.
5.	The second objective was to field verify the laboratory responses by
measuring the same responses 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.
9

-------
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 a historical site known as the Cen-
tral Long Island Sound (CLIS) disposal site (1.8 by 3.7 km) located approxi-
mately 15 km southeast of New Haven, Conn. (Figure 1). The sedimentology at
BRIDGEPORT
FVP STUDY
REACH
NEW
HAVEN
BRIDGEPORT
MAINTENANCE
(J DREDGING
BLACK ROCK
HARBOR
CLIS
black
ROCK
HARBOR'.
0 400m
LONG ISLAND
FVP
DISPOSAL
SITE
1 km
25 km
SOUTH REFERENCE
• SITE
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. Suspended sediment concentrations average 10 mg/Jl, with storm-
induced values to 30 mg11. The baseline community data revealed a homoge-
neous, mature infaunal community dominated by the polychaete Nephtys incisa
and the bivalve molluscs Nuaula proxima and Yoldia limatula.
8. The FVP disposal site was selected within the CLIS so as to minimize
contamination from other sources, including relic disposal operations or
10

-------
ongoing disposal activities occurring during the study period. This was nec-
essary to ensure a point source of contamination. The uniformity of physical,
chemical, and biological properties of the disposal site prior to disposal al-
lowed 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 OS
52.5 72
5
y
52.0 72
FVP DISPOSAL
- -
51.5 72
SITE
\
51.0
4 1 OS
/
/
/
/
/
/
I
V
\
s
V.
V,
i—i—r			i	1—i
0 500 m
).0
// ^ \
/ // 1
' ''
\ v V CNTR ' 20
	^,
-
REFS
3 Km
Y%
1 1
0E 400E
/ /
s'
•
I00OE





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 constituted a point source of contamination. The
temporal scale for the study was A years, which included a year of predisposal
data collection to define seasonal patterns in the physical, chemical, and
biological variables and 3 years of postdisposal data collection to address
the objectives of the program and to evaluate the long-term impacts of the
disposal operation on the surrounding benthic communities.
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
11

-------
was 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 6,400 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, respectively. Alkyl homologs of the PAHs
were also present in the dredged material at concentrations between 1,000 and
13,000 ng/g. Inorganic contaminants of toxicological importance present in
the dredged material included copper (2,900 yg/g), chromium (1,480 yg/g), zinc
(1,200 yg/g), lead (380 yg/g), nickel (140 yg/g), cadmium (24 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: (a) a suite of bio-
logical endpoints using the same material was developed and evaluated; (b) the
biological tests represented different levels of biological organization;
(c) the tests were conducted under both laboratory and field exposure condi-
tions; (d) the tissue residues were examined concurrently with measurements of
biological effects; (e) the duration of the study was adequate to evaluate the
use of community responses as a benchmark against which other biological re-
sponses could be compared; and (f) the project was a site- and waste-specific
case study for the application and evaluation of the components of a risk as-
sessment, including the development of methodologies for predicting and mea-
suring field exposures in the water column and benthic compartments. Limita-
tions of this study were: (a) only one dredged material was evaluated, which
constrained certain types of comparisons; (b) the size of the study put limits
on the extent to which any given objective could be examined; and (c) the
resources allocated to determine field exposures were limited. The latter
constraint was particularly important because the laboratory-field comparisons
and the risk assessment process both required accurate predictions of environ-
mental exposures.
12

-------
Laboratory-to-Fleld 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 the 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 labora-
tory systems. Consequently, the approach chosen for this program was to de-
velop laboratory exposure-response data using only general field exposure
information.
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 im-
ply 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 re-
sponses are being measured. Residue-effect relationships will be described
and interpreted for both laboratory and field exposures.
Adenylate Energy Charge
16.	Atkinson and Walton (1967) proposed adenylate energy charge (AEC)
as a measure of energy potentially available from the adenylate system for
13

-------
cell metabolism. The AEC defined as (ATP + 1/2 ADP)/(ATP + ADP + AMP),* has a
maximum value of 1.0 when all adenylate is in the form of ATP, and a minimum
value of 0 when all adenylate is in the form of AMP (Atkinson and Walton
1967). The energy charge is considered important in the control of key cata-
bolic and anabolic pathways (Atkinson 1971). Values of energy charge corre-
late with physiological condition: energy charges between 0.8 and 0.9 are
typical of organisms that are actively growing and reproducing, usually under
optimal environmental conditions (Atkinson 1971; Chapman, Fall, and Atkinson
1971; Rainer, Ivanovici, and Wadley 1979; Ivanovici 1980b). Values in the
range of 0.5 to 0.7 have been observed in organisms that are stressed (Ball
and Atkinson 1975; Behm and Bryant 1975; Wijsman 1976; Rainer, Ivanovici, and
Wadley 1979; Ivanovici 1980b) and whose growth and reproductive rates are re-
duced (Chapman, Fall, and Atkinson 1971). Values below 0.5 have been asso-
ciated with irreversible loss of viability under detrimental conditions (Ridge
1972; Montague and Dawes 1974; Skjoldal and Bakke 1978).
17. In this report the responses of adenine nucleotides and AEC to
stress are considered. The central role of adenine nucleotides in energy
transformation and in metabolic regulation suggests their potential usefulness
as indicators of sublethal stress. These responses were measured in the mus-
sel Mytilus edulis and the polychaete Nephtys inoisa exposed to a contaminated
dredged material, BRH sediments, both in the laboratory and in the field.
* ATP = adenosine triphosphate, ADP - adenosine diphosphate, and
AMP - adenosine monophosphate.
14

-------
PART II: MATERIALS AND METHODS
Laboratory Methods
Sediment collection
18.	Two sediment types were used to conduct laboratory tests for the
field verification studies. The reference (REF) sediment was collected from
the South Reference site in Long Island Sound (40ฐ7.95' N and 72ฐ52.7' W) by
2
Smith-Maclntyre grab (0.1 m ), press sieved through a 2-mm sieve, and stored
at 4ฐ C until used. Prior to dredging, contaminated sediment was collected
2
from BRH (41ฐ9' N and 73ฐ13' W) with a gravity box corer (0.1 m ) to a depth
of 1.21 m, thoroughly mixed, press sieved through a 2-mm sieve, and refriger-
ated (4ฐ C) in barrels until used. Details of sediment collection and storage
procedures may be found in Rogerson, Schimmel, and Hoffman (1985).
Organism collection and holding
19.	Mytilus edulis. Two separate experiments were completed using oxi-
dized REF and BRH sediments. Mussels were collected from the Narragansett Bay
reference population (71ฐ24.0' W by 41ฐ29.4' N) with a scallop dredge from a
depth of 10 m. Collection information for each experiment is listed in
Table 1. The animals 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 1ฐ C per day to 15ฐ C.
Table 1
Collection Information for the M. edulis Used in the
Laboratory Experiments
Collection Experiment Temperature
Experiment	Date	Begun	ฐC	
1	17 Jan 85	05 Feb 85	2.0
2	22 Feb 85	12 Mar 85	5.0
Salinity
g/k6
30.0
30.0
20. Nephtya inoiaa. Nephtys inaisa for laboratory studies were col-
2
lected with a Smith-Maclntyre grab sampler (0.1 m ) at the South Reference
site (Figure 1). Collection information for each experiment is listed in
Table 2. The sediment containing the N. inoiaa was brought to the laboratory
where it was sieved and the N. inoi8a were picked and sorted by size. Tests
15

-------
Table 2
Collection Information for the N. inoisa Used in the
Laboratory Experiments

Duration
Collection
Experiment
Temperature
Salinity
Experiment
days
Date
Start Date
ฐC
g/kg
1
10
30 Oct 84
10 Dec 84
15
28.0
2
28
27 Feb 85
12 Mar 85
1
28.5
3
42
23 Apr 85
3 May 85
10
29.3
were conducted with adult
specimens. These individuals
were placed
in REF
sediment,
in flowing seawater, and were
acclimated at a
rate of 1ฐ
C per day
to 20ฐ C.
They were fed
powdered prawn
flakes, ad libitum, during
this period
Suspended
sediment dosing system



21. Laboratory studies required the construction of two identical sedi-
ment dosing systems to provide simultaneously either BRH or REF material as
suspended sediment. Each dosing system (Figure 3) consisted of a conical-
shaped slurry reservoir placed in a chilled fiberglass chamber, a diaphragm
pump, a 4-ฃ separatory funnel, and several return loops that directed the
AROON
SEPARATORY
FUNNEL /
Figure 3. Suspended sediment
dosing system
WATER BATH
DELIVERY
MANIFOLD
DOSING
VALVE
TO EXPOSURE
SYSTEM
PUMP
RETURN
MANIFOLD
SLURRY
RESERVOIR
CHILLED
16

-------
particulate slurry through dosing valves. The slurry reservoirs (AO cm in
diameter by 55 cm high) contained 38 I of slurry composed of 36 I of filtered
seawater and 2 ฃ of either BRH or REF sediment. The fiberglass chamber (94 cm
by 61 cm by 79 cm high) was maintained between 4ฐ and 10ฐ C using an exter-
nally chilled water source to minimize microbial degradation during the test.
Polypropylene pipes (3.8 cm diam) extended to the bottom of the reservoir
cones and were connected to pumps (16- to 40-ฃ/min capacity) fitted with
Teflon diaphragms. These pumps were used to circulate the slurry while mini-
mizing abrasion that might produce changes in the physical properties (e.g.,
particle size) of the material.
22.	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 y) through sand filters was used.
Suspended sediment oxidation system
23.	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.
24.	In order to obtain consistent states of oxidation for both REF and
BRH sediments, 2 ฃ of sediment was transferred to an inverted polycarbonate
carboy and diluted to 19 i. 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
25.	Mytilue edulia. An exposure system was constructed to provide a
constant concentration of suspended sediment to mussels in the laboratory.
17

-------
2mm capillary tubing
utility strop
polycorbonaie
jugs
stand
polyethylene
tubing
Neoprene
stopper
polypropylene
volves 8 fittings
Figure 4. Suspended sediment oxidation system
This system consisted of recirculating loops from the suspended sediment
dosing system connected to a dosing valve at each exposure chamber. The con-
centration 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 cham-
ber 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 concentrations of BRH and REF sediment varied between treatments; how-
ever, a total suspended sediment concentration of approximately 10 mg/#. (dry
weight) was maintained in all five laboratory exposure treatments. This con-
centration was chosen because it approximated the background field suspended
sediment concentration present at the CLIS disposal site.
26. 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 sus-
pended sediment concentrations, measured by filtration onto glass fiber fil-
ters, with the transmissometer units displayed on a microprocessor. A trans-
missometer value was calculated that corresponded with the desired suspended
18

-------
seawater/seqiment
SLURRY
ALGAE
Figure 5. Laboratory exposure
system for M. edulis
TRANSMISSOMET
OR
TO MICROPROC
MIXING
CHAMBER
AIR STONE
sediment concentration of 10 mg/ฃ for each chamber. As the mussels removed
suspended sediments, the microprocessor opened dosing valves to deliver addi-
tional suspended sediment at 2-min intervals. In this manner, suspended sedi-
ment concentrations were maintained at the desired values (tlO 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 oxy-
gen and to ensure even distribution of suspended particulates (Figure 5).
27.	In addition to the suspended sediment, food in the form of a uni-
cellular alga, Isoehrysis 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/ฃ. 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 15ฐ 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.
28.	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 as-
sess the biological effect on these organisms. Mytilus eduHs were exposed
for approximately 1-month periods at the CLIS disposal site; therefore,
19

-------
exposures of similar duration, 28 days, were used for the laboratory exposures.
29.	At the start of both experiments, 150 mussels were placed into each
chamber. MytiZus edulis were sampled at time zero for determination of ini-
tial tissue residue concentrations and for adenine nucleotide measurements.
30.	Experiment 1 consisted of three exposure treatments: 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 was
terminated at day 14 because adverse biological effects (e.g., reduced filtra-
tion rate) were observed in both treatments containing BRH sediment.
31.	Experiment 2. was conducted with lower concentrations of BRH sus-
pended sediment. Exposure treatments of suspended sediment in Experiment 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 particulate water
samples were taken for chemical analysis 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 chamber.
32.	The operation of the system (dosing valves, flow rates, etc.) was
monitored daily. Experiments using the 100- and 0-percent BRH treatment re-
quired only one dosing valve each, while the 50-percent BRH treatment required
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.
33.	Nephtys inciea. In the laboratory tests with N. inaisa, the dosing
system was set to maintain nominal concentrations of 200 mg/Jl (dry weight) of
suspended sediments with seawater flow rates producing five volume replace-
ments per exposure chamber per day. These flow rates meet the minimum recom-
mended by the American Society for Testing and Materials (1980) and were in-
tended to maximize residence time of the suspended sediments in the exposure
chambers.
20

-------
34. A suspended sediment proportional diluter (Figure 6) was used to
mix the small quantities of concentrated sediment slurries (10 to 20 g/ฃ from
the sediment dosing system with filtered seawater to produce dilute sediment
suspensions in the milligrams-per-litre range. It then combined slurries of
different types (e.g., REF and BRH sediment suspensions) proportionally to
maintain the same concentration of suspended sediment with different ratios of
the two sediments.
(from 3-way valves)
BRH slurry REF slurry
solenoid
valve
seawater
counter
relay switch
^-manifold
water
ce
water
trap
i-float chamber
mixing
chamber
, float switch
[ft-*--
y
distribution
chambers
I lection
chambers
splitter
Figure 6. Proportional diluter used to deliver
suspended sediment to the N. inaisa exposure
chambers
35. The exposure chamber for N. incisa is illustrated in Figure 7.
Polycarbonate bottles (19 I) used commercially for shipping spring water were
cut off at the top. REF sediment (2 Jl/chamber) was added to a depth of 4 cm,
and Plexiglas strips were inserted into the sediment, dividing it into pie-
shaped sections. This permitted subsampling without disturbing the entire
chamber. Each chamber was filled with filtered seawater at 20ฐ C. After the
sediment in the chambers was permitted to settle and equilibrate for about
21

-------
DOSING CHAMBER
delivery f
from dosi
system
air lift for
circulation
Figure 7. Nephtys incisa
exposure chamber
REF ^
sediment —
	L
water
both (200C)
\
Plexiqlas partition
4 hr, N. incisa were added, and an additional 2 hr was allowed for the worms
to burrow into the sediment. The delivery tubes from the proportional dilute
were then put in place, and a low pressure airlift was turned on to keep the
dosed sediments in suspension. This system allowed very little sediment depo
sition during the course of experiments. Excess seawater was permitted to
overflow the brim of each chamber. Earlier experiments indicated that once
the worms burrowed into clean REF sediment, they would not attempt to escape.
Therefore, the chamber design used here was considered acceptable. Two cham-
bers were used for each of the three treatments for a total of six chambers
per treatment. The two chambers did not represent replicates, but were used
to accommodate enough worms for chemical and biological analysis in each
experiment.
36.	Three experiments were conducted during this phase of the FVP in
which adenylate nucleotide measurements were made. These experiments lasted
10, 28, and 42 days, respectively, and had exposure conditions of 100-, 50-,
and 0-percent BRH suspended sediment. The 42-day experiment provided time
series sampling for the three exposure conditions. Worms were removed at
time 0, day 28, and day 42. This experiment was supported with chemical anal
yses of the seawater and of the N. incisa. Nephtys incisa were collected on
sieve after removal of a pie-shaped aliquot of bedded sediment from each cham
ber. Clean REF sediment, without N. incisa, was returned to the vacated sec-
tion to maintain the integrity of the exposure chamber.
37.	Suspended sediment, temperature, and salinity were measured rou-
tinely during each experiment. Dissolved oxygen (DO) concentrations were not
expected to be a problem because of the large volume of the chamber and the
22

-------
use of an airlift. However, DO levels were determined once during each exper-
iment and never differed significantly from expected saturation levels. The
worms were fed 100 mg of powdered prawn flakes per chamber per day for the
duration of each experiment.
Adenylate nucleotide extraction
38.	Mytj'Zus edulis. The adductor muscle was rapidly dissected out,
blotted dry, placed on a labelled polyethylene strip (Gladwrapฎ) , and freeze
clamped with aluminum blocks cooled to -196ฐ C with liquid nitrogen (Bergmeyer
1965; Ivanovici 1980a). The time between sampling and dissection never ex-
ceeded 10 min. Tissue samples were removed and freeze clamped in less than
30 sec, and the labelled samples were stored in liquid nitrogen until
homogenization.
39.	Adenine nucleotides were extracted from tissues with a method simi-
lar to that of Ivanovici (1980a) (Figure 8). The freeze-clamped tissue was
quickly transferred from its wrapping to a tared stainless steel homogenizing
tube previously cooled in liquid nitrogen and placed in a polyurethane insula-
tor and weighed. Tissue samples (approximately 0.2 g) were ground to a fine
powder at -196ฐ C. Perchloric acid (PCA) (1 ml, 6 percent v/v) was added to
the ground tissue, allowed to freeze, ground to a powder, and mixed with the
tissue sample. This mixture was kept on ice and allowed to thaw, after which
additional ice-cold PCA was added (the final ratio of tissue to PCA was 1:10,
w/v) and then centrifuged at 5ฐ C and 6,000 g's for 20 min after thorough mix-
ing. The supernatant was decanted into a (polyethylene) centrifuge tube con-
taining 5 of universal indicator and adjusted to pH 6.5 to 7.0 with solid
K^CO^. These tubes were left on ice for approximately 15 min to allow CO^
evolution and then centrifuged as above. The supernatant was decanted from
the KCIO^ precipitate into clean (polyethylene) centrifuge tubes and assayed
or stored at -20ฐ C. Generally, 20 samples were prepared each day. Extrac-
tion efficiencies of the adenine nucleotides from adductor tissue muscle tis-
sue of M. edulis by PCA were consistently greater than 92 ฑ 0.5 percent.
40.	Nephtys ineisa. Worms collected on a fine mesh sieve (0.9-mm mesh)
were immediately anesthetized by immersion of sieve and worms into a 7-percent
solution of MgCl2 in seawater for 2-1/2 min (Dean and Mazurkiewicz 1975). The
worms were washed by immersion of the sieve in clean seawater, and the worms
were then removed from the sieve and placed into a Carolina dish (75 mm diam)
containing approximately 50 ml clean seawater. One or two anesthetized worms
23

-------
SAMPLE: ADDUCTOR MUSCLE TISSUE |>0.2gWETWT)
FREEZE CLAMP IN POLYTHENE FILM
STORE IN LIQUID N2
~
WEIGH, HOMOGENIZE IN LIQUID N2,
ADD 1 mS6% PCA (v/v)
~
ADD MORE 6% PCA (TISSUE: ACID, 1:10}
STAND FOR APPROXIMATELY 15 MIN @ 0ฐ C
t
CENTRIFUGE 20 MIN, S,OOOg's@5ฐC
i	)
SUPERNATANT:	DISCARD PELLET
ADJUST TO pH 6.5-7.0
WITH SOLID K2C03
I
CENTRIFUGE 20 MIN, 6,000 g's <ง> 5 C
i
f	t
DISCARD PELLET	NEUTRALIZED SUPERNATANT,
ANALYZE ATP, ADP. AMP
OR FREEZE AND STORE @ - 20ฐ C
Figure 8. Summary of procedure for the
extraction of adenine nucleotides from
the adductor muscle of M. edulis
(=>0.1-g wet weight) were placed on a millipore filter pad (25 mm, 1.2 v)> and
as much seawater as possible was removed by vacuum. The anesthetized worms
were gently removed from the filter pad onto a labelled polyethylene strip
(Gladwrapฎ) and freeze clamped with aluminum blocks cooled by nitrogen to
-196ฐ C (Bergmeyer 1965; Ivanovici 1980a).
41, Adenine nucleotides were extracted from tissues with a method simi-
lar to that of Ivanovici (1980a) (Figure 9). The freeze-clamped tissue was
quickly transferred from its wrapping to a tared stainless steel homogenizing
tube previously cooled in liquid nitrogen and placed in a polyurethane insula-
tor and weighed. Tissue samples (approximately 0.1 g) were ground to a fine
powder at -196ฐ C. One half the total volume required of PCA (6 percent v/v)
containing 0.33 percent ethylenediamine tetra-acetic acid (EDTA) (w/v) was
24

-------
SAMPLE: ONE WORM (3-4 CM; >0.1 g WET WEIGHT)
FREEZE CLAMP IN POLYTHENE FILM
STORE IN LIQUID N2
WEIGH, HOMOGENIZE IN LIQUID N2,
ADD 1/2 TOTAL VOLUME OF 6% PCA (v/v) - 0.33% EDTA (w/v)
REQUIRED TO MAKE A FINAL DILUTION OF 1:10 (TISSUE: ACID)
THAW @ 0ฐ C
CENTRIFUGE 20 MIN, 6.0O0g's  5ฐ C

PELLET	SUPE RNATANT(1)
ADD 1/2 THE TOTAL VOLUME
OF 6% PCA (v/v| - 0.33% EDTA (w/v)
AS ABOVE, SONICATE 30 SEC
I
CENTRIFUGE 20 MIN, 6,000 g's @ 5ฐ C
H	1
PELLET (DISCARD)	SUPERNATANT (2)
COMBINE SUPERNATANTS 1 AND 2
ADJUST TO pH 6.5-7.0 WITH
SOLID K2C03
I
FILTER THROUGH MILLIPORE
SWINEX FILTER (0.45n. 13 MM)

NEUTRALIZED SUPERNATANT
ANALYZE ATP, ADP, AMP WITHIN
2 HR
Figure 9. Summary of procedure for the extraction of
adenine nucleotides from the tissue of N. inai.8a
added to the ground tissue, allowed to freeze, and ground to a powder with the
tissue sample. This mixture was kept on ice and allowed to thaw. After thaw-
ing, the mixture was centrifuged at 5ฐ C and 6,000 g's for 20 min after
thorough mixing. The supernatant was decanted into a polyethylene centrifuge
tube. Each pellet was extracted again with one half the total volume of PCA
(6 percent v/v) - EDTA (0.33 percent w/v) required to make a final dilution of
1:10 (tissue:acid) and followed by sonication and centrifugation. The
25

-------
supernatants were combined, and 5 uฃ of universal indicator was added and ad-
justed to pH 6.5 to 7.0 with solid K^CO^. These tubes were left on ice for
approximately 15 min to allow CO^ evolution and then centrifuged as above.
The supernatant was decanted from the KCIO^ precipitate into clean polyethyl-
ene centrifuge tubes and assayed or stored at -20ฐ C. Generally, 20 samples
were prepared each day. Recovery efficiency of the extraction was determined
by spiking tissue samples with ATP, ADP, and AMP, and recovery was calculated
by the following equation:
% Recovery = (Sample + Standard) - Sample x 10Q
Standard
where
Sample + Standard = concentration of adenylates in sample spiked with
adenylates
Sample = concentration of adenylates in samples
Standard = concentration of adenylate standard
Adenylate assay
42.	Mytilus edulis. The concentrations of ATP, ADP, and AMP were de-
termined spectrophotometrically (340 nm) with hexokinase (Lamprecht and
Trautschold 1974), pyruvate kinase, and myokinase (Adam 1963), respectively
(Figure 10). All enzymes, chemicals, and reagents (analytical grade) were ob-
tained from Boehringer Mannheim, Indianapolis, Ind.
43.	The principle of the ATP assay is as follows: glucose is phos-
phorylated by ATP to glucose-6-phosphate (G6P) with hexokinase (HK)
(reaction 1). Glucose-6-phosphate then reacts with nicotinamide-adenine
dinucleotide phosphate (NADP) to form 6-phosphoglucono-lactone and reduced
nicotinamide-adenine dinucleotide phosphate (NADPH). This reaction is cata-
lyzed by glucose-6-phosphate dehydrogenase (G6P-DH) (reaction 2).
HK
REACTION 1: ATP + GLUCOSE	G6P + ADP
Mg2+
REACTION 2:
G6P + NADP—
6-PHOSPHOGLUCONO-LACTONE
"+ NADPH + H+
26

-------
NEUTRALIZED SUPERNATANT
(SEE FIGURES 8 AND 9)
f
i
ATP ASSAY
I
0.2 mC INTO CUVETTE
1.8 mfi ASSAY BUFFER
5.0 8 G6P-DH, MIX
READ (340 nm)
5.0 2 GLUCOSE, MIX
IMMEDIATELY ADD
5.0 e HK, MIX
READ (340 nm) AFTER
20-MIN INCUBATION
LEGEND
G6P-DH ป GLUCOSE-6-PHOSPHATE DEHYDROGENASE
HK = HEXOKINASE
LDH - LACTATE DEHYDROGENASE
PK - PYRUVATE KINASE
MK = MYOKINASE
ADP, AMP ASSAY
0.2 mfi INTO CUVETTE
1.8 mC ASSAY BUFFER
5.0 6 LDH, MIX
READ (340 nm)
5.0 E PK, MIX
READ (340 nm) AFTER
20-MIN INCUBATION
5.0 e MK, MIX
READ (340 nm) AFTER
20-MIN INCUBATION
Figure 10. Summary of procedures for analysis of ATP, ADP, and AMP
in adductor muscle tissue of M. edulis and whole N. inaisa
Thus for every micromole of ATP, 1 ymol of NADPH is formed and causes an in-
crease in absorbancy at 340 nm.
44. The principle of the ADP and AMP assays is as follows: pyruvate
kinase (PK) catalyzes the phosphorylation of 1 ymol of ADP by phosphoenolpyru-
vate (PEP) to form 1 ymol of ATP and pyruvate (reaction 4). Pyruvate in turn
27

-------
is converted to lactate by lactate dehydrogenase (LDH). Thus, 1 ymol of ADP
results in the formation of 1 ymol of nicotinamide-adenine dinucleotide (NAD)
(reaction 5). The decrease in absorbancy at 340 ran caused by the formation of
NAD from NADH is, therefore, proportional to the amount of ADP present in the
sample. After this absorbance change has been measured in a sample, myokinase
(MK) is added. This enzyme catalyzes the formation of 2 ymol of ADP from
1 ymol each of AMP and ATP (reaction 3). In turn, 2 ymol of NAD are formed
(reactions 4 and 5).
MK
REACTION 3: AMP + ATP
2 ADP
REACTION 4: 2 ADP + 2 PEP-
2 ATP + 2 PYRUVATE
REACTION 5:
2 PYRUVATE + 2 NADH	
2 LACTATE + 2 NAD
45. To determine if any inhibitory effects of neutralized tissue
extracts on the nucleotide assay system occurred, known amounts of ATP, ADP,
and AMP were added to neutralized extracts as internal standards and assayed
to check for inhibitory or enhancement effects by the extract. The following
equations were used to calculate correction factors (Cf):
X7 = (Sample + Internal Standard) - Sample
Internal Standard
(2)
Cf	= 100Z
ATP, ADP, or AMP 100% + X%	K '
46. A Cf was not required for ATP since extracts of M. edulis adductor
muscle had a negligible effect on absorbance. However, these same extracts
increased absorbance, which caused high readings for ADP (112 percent) and AMP
28

-------
(Ill percent). Thus, a CF was required for ADP (0.89) and AMP (0.90) to cal-
culate accurately their concentration.
47.	Nephtys inoisa. Within 2 hr after the final centrifugation,
N. inoisa tissue extracts were assayed for adenylate using a technique identi-
cal to that for M. edulis (Figure 10).
Field Methods
Organism collection and holding
48.	Mytilus edulis. 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 returned to the laboratory where 100, 5- to 7-cm organisms were
sorted and placed into each polyethylene basket. All baskets were placed in
holding tanks of flowing unfiltered seawater until deployed in the field.
49.	Nephtys inoisa. Nephtys inoisa for field studies were collected at
stations REFS, 1000E, 400E, 200E, and CNTR. Station locations were marked
with buoys for the duration of this project. While the boat was anchored, a
2
Smith-Maclntyre grab sampler (0.1 m ) was used to collect bottom sediments.
These sediments were wet sieved on deck (nested sieves of 2- and 0.5-mm mesh
size), and organisms were collected. On each sampling date, N. inoisa were
collected for biological measurements. Specimens for adenylate measurements
were prepared immediately on the boat.
Exposure
50.	Mytilus edulis deployment and retrieval. Mytilus edulis were de-
ployed at CNTR, 400E, 1000E, and REFS at the CLIS disposal site (Figure 2).
The physical arrangement at 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 at-
tached 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.
51.	Deployment of M, edulis at the CLIS disposal site is summarized in
Table 3. Mussels were deployed at each station for a period of 1 month pre-
disposal to collect baseline data (cruise number T - 4). A second deployment
occurred during disposal operations, except that no mussels were placed at the
29

-------
Table 3
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
-
4
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
3
months
T
+
55
18
Oct
83
05
Jun
84
8
months
T
+
74
12
Jun
84
17
Oct
84
4
months
* T = 0 refers to the termination of disposal activities at the FVP site on
18 May 1983.
CNTR station (T = 0, T + 2). Mussels were deployed for 1-month periods over
the next 3 months (T +8, T + 12, T + 16) and then on a quarterly basis for
the next year (T + 23, T + 40, T + 55, and T + 74). In addition, several sets
of mussels were left at each station for 7 months (T + 22).
52.	Mytitus 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.
53.	Mytilus edulis field exposures via tissue residues. Exposure con-
ditions present in the field during each mussel deployment were not as well
characterized as they were in the laboratory studies. As a result, the de-
scription of M. edulis exposure to BRH material in the field is more qualita-
tive than quantitative and will be presented in two parts. First, a
30

-------
prediction of field exposure is based on mussel tissue residues. The rela-
tionship between exposure 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 expo-
sure concentrations (0, 1.5, 3.3 mg/il) from the same exposures. In order to
correct for background 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, mg/il BRH material = (PCB residue
x 0.000965) - 0.0019, (R^ ฆ 0.99) , 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
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 that mussels were retrieved.
54.	Mytilus edulis estimated exposure via water chemistry data. A sec-
ond estimate of exposure was generated from the PCB and copper concentrations
in the whole water samples collected during the various postdisposal cruises.
The concentration of BRH material that would have to be present to produce
these levels was determined by dividing the concentration of PCB and copper
present in the barrel material collected from BRH (2,900 ug/g and 6,910 pg/g
for copper and PCB, respectively). A range of exposures also was calculated
for the water chemistry data; estimated BRH material was determined with and
without subtracting the concentration at the REFS station.
55.	Nephtys inaisa field exposures via tissue residues. The purpose of
exposure assessment is to determine the temporal and spatial range of exposure
concentrations experienced by populations of interest. The exposure condi-
tions present in the field for N. inaisa were not as well characterized as
they were in the laboratory studies. As a result, the description of
N. inaisa exposure to BRH material in the field is more qualitative than quan-
titative and is presented in three parts. First, a prediction of field expo-
sure can be made based on worm tissue residues. The relationship between
31

-------
exposure to BRH sediments and tissue residues was determined in the laboratory
experiment. Tissue residues of PCBs as Aroclor 1254 (A1254) from the 0-, 50-,
and 100-percent BRH treatments at 42 days were plotted against BRH exposure
concentrations. This relationship was used to estimate field exposure condi-
tions based on tissue residues of PCBs in field-collected worms. Inherent in
this approach is the assumption that organisms have comparable patterns of
bioaccumulation in the laboratory and in the field.
56.	Nephtys inoisa field exposures from physical data. A second analy-
sis calculates the maximum total suspended solids concentrations from 1 m
above the bottom to the sediment-water interface. This analysis assumes that
the total suspended solids are composed totally of BRH sediments and repre-
sents a worst case or upper bound prediction. A third approach calculates the
probable amount of BRH sediment exposure at the sediment-water interface based
upon actual sediment contaminant concentrations for each sampling station and
date. This analysis assumes that resuspension of the surface sediment is the
primary source of the total suspended solids at the sediment-water interface.
57.	The equation used to calculate total suspended solids concentra-
tions from the sediment-water interface up to 1 m above the bottom is de-
scribed as follows:
Cz - C„[' + 
-------
factors were likewise empirically determined from acoustic profilometer data
collected between the sediment-water interface and 1 m above the bottom
(Bohlen and Winnick 1986; Munns et al. 1986) .
Maximum upper bound estimate
59.	For the purposes of the maximum upper bound analyses, it was as-
sumed that the exposed populations are located off the mound and aligned with
the mean direction of current flow. The route of contaminant exposure was as-
sumed to be through the transport of resuspended BRH sediments. These total
suspended solids are composed of resuspended Long Island Sound sediments, as
well as BRH sediment resuspended from the disposal site. Since the intent of
these analyses is to create a maximum upper bound set of exposure conditions,
it was assumed that the suspended solids concentration was composed, in total
(100 percent), of resuspended BRH sediment.
Probable exposure estimate
60.	It was not within the scope of this program to provide a continuous
temporal record of the percent contribution of BRH sediments to the total sus-
pended solids load. Consequently, a second set of analyses was designed to
estimate the percentage of BRH sediment that could have comprised the total
suspended solids concentration at the sediment-water interface for each sta-
tion and indicate how these concentrations changed with time throughout the
study. The proportions of BRH dredged material in the surficial sediments at
each station and date were estimated by comparing the concentrations of
selected contaminants measured in the 0- to 2-cm layer of sediment cores col-
lected, postdisposal, at the FVP site. These field concentrations were com-
pared with the barrel concentrations to determine a percentage as follows.
Percentage BRH Sediment
where
(C - REF)
(BRH - REF)
x 100	(5)
C * concentration of contaminant in the sediment core
REF - concentration of contaminant in REF sediment
BRH ซ concentration of contaminant in BRH sediment (barrel)
The percentage BRH sediment values were calculated for each station and date
using the 11 different contaminants, the details of which are shown in Appen-
dix A, Tables A1-A13. To achieve a BRH-suapended sediment concentration that
reflects the surficial sediment contaminant levels for each station and date,
33

-------
the total suspended solids concentrations predicted for the sediment-water
interface were multiplied by the estimated proportions of BRH sediment.
Chemical Methods
Analytical methods
61.	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 in order to ana-
lyze the types of matrices in this study. These methods were intercalibrated
to ensure the quality of the data.
Organic sample preparation
62.	Samples of sediment, suspended particulates, and organisms were ex-
tracted 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 were saved. Foam plugs containing the
dissolved 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 re-
duced carefully prior to analysis.
Organic analysis
63.	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 A1254 standard be-
cause the distribution of PCB congeners in the dredged material closely
matched that distribution, as did the distribution in organisms at
steady-state.
64.	Gas chromatograph/mass spectrometric analyses were conducted with a
Finnigan Model 4500, also equipped with a 30-m DB-5 fused silica capillary
column. The mass spectrometer was operated through a standard Incos data
34

-------
system and was tuned at all times to meet USEPA quality assurance
specifications.
65.	All instruments were calibrated daily with appropriate standards.
The concentrations of the standards used were chosen to approximate those of
the contaminants of interest, and periodic linearity checks were made to en-
sure the proper performance of each system. When standards were not avail-
able, 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 reduction
66.	As stated above, PCBs were quantified as A1254 because the sample
patterns closely resembled that profile. This allowed a convenient way of re-
porting these data without treating the voluminous data that would have re-
sulted from measuring some 55 congener peaks by electron capture detector.
Likewise, a method was sought to summarize the PAH data. Appendix B lists the
35 individual PAH parent and alkyl homolog compounds and groups of compounds
measured in this study. Each PAH of the same molecular weight, both parents
and alkyl homologs, can be summed to yield 9 PAH parent sums and 5 alkyl
homolog sums. Although useful, this only reduced the data to 14 PAH vari-
ables, which was not sufficient. Since the distribution of PAHs differed
greatly in both quantity and quality between Long Island Sound and the BRH
dredged material, statistics were sought that would retain significant quanti-
tative 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 - S[C(i)]
(6)
CENT -
S[C(i) * MW(i)]
SUM
(7)
where
S - SUM
C(i) ป concentration of PAH from molecular weight 166 through 302,
including both parent and alkyl homologs
35

-------
t- vป
MW(i) = molecular weight of 1 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
molecular weight of any particular PAH distribution. This statistic can be
used to readily distinguish two different sources of PAH distributions; one
with predominately heavy molecular weight pyrogenic compounds and one with
more lighter molecular weight petrogenic compounds. These distributions are
typically found in Long Island Sound at REFS and BRH, respectively. A major
value to this statistic is that it enables one to readily distinguish these
two sources when their contaminant concentrations are nearly equal. The for-
mulas for calculating these, and 178 alkyl homologs, are shown in Appendix B.
Because distributions of both parents and homologs were measured, SUMs and
CENTs of both parents and homologs were calculated as well. These were de-
fined as PSUM, PCENT, HSUM, and HCENT. By definition,
SUM = PSUM + HSUM	(8)
and
PSUM * PCENT + HSUM * HCENT
CENT = 	—		(9)
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
67.	Sediment was prepared for inorganic analysis by elution at room
temperature with 2N HNO^. The samples were filtered through Whatman it2 filter
paper. Organisms were totally digested in concentrated HNO^ at 60ฐ C and fil-
tered through Whatman //2 filter paper.
68.	Cadmium, nickel, lead, 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, and
zinc) were analyzed by heated graphite atomization atomic absorption (HGA-AA)
via direct injection. Samples of suspended particulates on Nucleopore
(0.45 y) filters were eluted with 2N HNC>3 and analyzed by HGA-AA.
Inorganic analysis
69.	All flame atomization atomic absorption (FA-AA) was conducted with
a Perkin-Elmer (Model 5000) atomic absorption spectrophotometer. All HGA-AA
36

-------
determinations were conducted with Perkin-Elmer Model 500 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.
70.	The FA-AA and HGA-AA instrument operating conditions are similar to
those described in USEPA (1979) and those in the manufacturers' reference man-
uals. The AA instruments were calibrated each time samples were analyzed for
a given element. Sample extracts were analyzed a minimum of twice to deter-
mine 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
71.	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. In determin-
ing the acceptability of dredged material for ocean disposal, a variety of
evaluatory criteria are applied. These include bulk sediment chemistry, tox-
icity, and bioaccumulation. 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 contami-
nants selected for detailed analyses throughout the study included PCBs, PAHs,
the pesticide ethylan, and eight metals.
72.	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.
73.	Multivariate clustering analyses were performed on the chemical
data in an attempt to define groups or clusters of chemicals that behaved in a
statistically similar manner. No assumptions were made concerning the behav-
ior, interactions, or dynamics of chemicals between compartments; therefore,
each compartment was analyzed separately. Five compartments were identified
37

-------
from field and laboratory data for statistical analysis. Of these, the surfi-
cial sediments and the unfiltered, particulate, and dissolved water column
fractions described exposure conditions experienced by infaunal and pelagic
organisms. The remaining compartment consisted of tissue residues in
organisms.
74.	The data were further partitioned into inorganic and organic analy-
sis. The inorganic analyses generally consisted of 8 variables, whereas the
organic 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. The subset of chemi-
cals selected as representative included six organic compounds, four metals,
and two summary statistics.
Statistical Analysis Methods
75.	The primary objective of the FVP was to compare laboratory with
field responses under similar exposure conditions. Because of the highly
dynamic temporal and spatial conditions in the field, the exposure environment
can be given only boundaries and cannot be assigned specific values, as is the
case for laboratory studies. Consequently, the degree to which laboratory
exposure-response relationships concur with those derived from field data can
be described only qualitatively. That does not preclude the use of inferen-
tial statistical procedures to explore those laboratory and field relation-
ships for which the appropriate quantitative information is available. How-
ever, the nature of this project was such that descriptive and exploratory
statistics were often the most appropriate techniques to illustrate relations
and trends. Simple graphic representations of variables were all that was
necessary to illustrate a relationship. In addition, multivariate techniques,
such as cluster analysis, were the most appropriate techniques to elucidate
more complex relationships between groups of selected variables.
76.	Prior to making comparisons between laboratory and field effects,
it was necessary to establish whether field exposure boundaries were similar
to those measured in the laboratory. Assuming that tissue residue and expo-
sure are closely related, this was accomplished by examining the tissue resi-
dues of all worms from laboratory and field exposures together, independent of
exposure concentration or station location and date. An agglomerative
38

-------
hierarchical cluster analysis was performed on the ten selected chemical con-
taminants and the two summary statistics using the SAS cluster procedure (SAS
1985) to establish which tissue residues among all the laboratory treatments
and field stations were most similar. The clustering procedure used was the
average linkage method, which uses unweighted pair-groups with arithmetic
averages on squared distances between samples. Prior to analysis, residue
data were normalized using standard deviations 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.
77. The relationship between AEC and tissue residue values in the field
samples was explored by regressing and plotting the mean AEC value for each
sample against the corresponding mean tissue residue (Snedecor and Cochran
1980). This procedure was completed individually for each of the 10 selected
chemical contaminants and the two summary statistics.
39

-------
PART III: RESULTS
Laboratory
Exposure
78. Mytilus edulis system monitoring. The M. edulis exposure system
was monitored for both total suspended solids (TSS) concentrations and the
percentage of REF and BRH sediments. A strip chart record indicated that the
system maintained a suspended particulate concentration of 10 mg/i approxi-
mately 90 percent of the time. Examples of times when the 10 mg/ฃ 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 con-
centration of BRH sediments dosed into each treatment is listed in Table 4.
Table 4
Suspended Sediment Concentrations in the Mussel Exposure System


Calculated
Nominal
Measured
BRH Sediment
% BRH
% BRH
mp/i,
100
100(0.0)*
10.0
50
50(0.83)
5.0
30
33(0.84)
3.3
10
15(1.39)
1.5
0
0(0.0)
0.0
* Standard error in parentheses.
79.	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 (ฑ5 percent) of BRH and REF material were delivered to the 50-percent
BRH exposure chamber. The 100- and 0-percent BRH treatments were controlled
by single dosing values.
80.	The 10- and 30-percent BRH treatments required two dosing valves
per treatment. Because the pulse length could not be adjusted separately for
each valve, manual adjustment of each valve was required to provide the de-
sired concentration. The volumetric amount of BRH and REF material delivered
40

-------
to each treatment was .onitored and recorded. In the treatment with a nominal
10-percent BRH, the actual value delivered was 15 percent. In the 30 percent
BRH treatment, the actual value was 33 percent.
81.	Mvtilus edulis 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 ซg/i in the exposure chambers. Initially, 150 animals were placed into
each chamber with clearance rates of approximately 2 */mussel/hr, or a total
of 300 4/hr. The seawater flow rate through each chamber, independent of sus-
pended sediment additions, was approximately 24 l/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 consequences on the behavior of the contaminants in the exposure
system.
82.	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 measured chemical concentrations in the expo-
sure system, using PCB and copper as examples.
83.	Whole water samples were taken for chemical analysis on days 1,7,
14, 21, and 28 in the second experiment. The mean PCB concentrations (nano-
grams per litre) for the five sampling dates for each exposure treatment in
the second experiment are given in Table 5. 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 ex-
posure system into the equation provided an estimated value of the concentra-
tion of BRH sediment in the system. The estimated concentration of BRH sedi-
ment in each treatment is similar to the actual measured values. These data
41

-------
Table 5
Chemical Monitoring of the Exposure System in Experiment 2
Nominal




Treatment




Concentration
PCB Concentration, ng/ฃ
BRH Concentration, mg/Jl
% 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
suggest that PCB concentrations in the system are closely associated with the
TSS concentrations.
84.	Copper concentrations were measured both with and without mussels
in the exposure system at 10 mg/ฃ TSS for each treatment. With no mussels in
the exposure system, the total copper concentrations were 9.37 and 2.5 yg/ฃ
for the 30- and 10-percent BRH treatments, respectively. These concentrations
represent 3.8 and 1.8 mg/4. 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.
85.	When mussels were present in the system, the mean copper concentra-
tions were 17.0 and 10.7 \ig/Z 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
suspended solids were delivered at a higher rate to the exposure chamber than
the rate of incoming seawater, soluble copper accumulated in the exposure
chamber. When a dose of BRH suspended sediment was delivered to an exposure
chamber, all contaminants were introduced at the same rate. Because the mus-
sels were more efficient at removing particulates than dissolved contaminants,
dissolved copper tended to accumulate because its removal from the system was
primarily via the overflow at a much slower rate. This resulted in higher
concentrations of copper than those predicted from the TSS data alone.
86.	Nephtys inoisa system monitoring. During the three N. incisa labo-
ratory experiments, the exposure system was monitored for TSS, seawater
42

-------
temperature, and seawater salinity. These data are presented in Table 6. In
general, the exposure system maintained the suspended solids concentrations
close to the nominal 200 mg/JL Temperature and salinity values were stable at
approximately 20ฐ C and 30 g/kg, respectively. DO concentrations were checked
once during each experiment, and they never differed significantly from ex-
pected saturation.
Table 6
Measured TSS Concentrations (Dry Weight) and Exposure
Conditions for Laboratory Tests with N. incisa


Seawater
Seawater
Treatment
TSS, mg/ซ,
Temperature
Salinity, g/kg
% BRH
x ฑ SD
x ฑ SD
X
+
SD

Test 1 -
10 Dec 1984 (10 days)



100
215 ฑ 53
19.9 ฑ 0.30
31.5
+
0.70
75
182 ฑ 14
19.9 ฑ 0.30
31.5
+
0.70
50
180 ฑ 19
19.9 ฑ 0.30
31.5
+
0.70
25
189 ฑ 22
19.9 ฑ 0.30
31.5
+
0.70
0
190 ฑ 16
19.9 ฑ 0.30
31.5
+
0.70

Test 2 -
12 Mar 1985 (28 days)



100
183 ฑ 24
18.9 ฑ 0.43
31.1
+
0.86
50
185 ฑ 21
18.9 ฑ 0.43
31.1
+
0.86
0
203 ฑ 24
18.9 ฑ 0.43
31.1
+
0.86

Test 3 -
3 May 1985 (42 days)



100
201 t 23
19.8 ฑ 0.53
30.9
+
0.70
50
184 ฑ 19
19.8 ฑ 0.53
30.9
+
0.70
0
190 ฑ 21
19.8 ฑ 0.53
30.9
+
0.70
87. Nephtys incisa chemical monitoring. During the 42-day experiment,
seawater and N. incisa from the exposure chambers were sampled for chemical
analysis. Seawater chemical monitoring data are presented in Table 7. The
dosing system malfunctioned for 2 days spilling BRH sediments into all treat-
ments. The day 18 chemistry samples were taken during this period. The prob
lem was corrected, and the system performed normally for the remainder of the
test. The seawater chemistry data confirm that N. incisa received a graded
exposure to BRH sediments during most (AO of 42 days) of the experiment.
43

-------
Table 7
Chemical Analysis of Seawater in Exposure Chambers of 42-Day
Experiment Exposing N. inaisa to BRH sediment
Experiment
Treatment
Total PCB
Total
Metals,
yg / ฃ
Day
% BRH
ng/ฃ as A1254
Cu
Cd
Cr
3
100
NS*
407
5.4
245

50
NS
256
3.2
159

0
NS
15
0.1
15
6
100
1,170
NS
NS
NS

50
590
NS
NS
NS

0
79
NS
NS
NS
18**
100
340
307
3.6
181

50
510
208
3.5
125

0
700
134
2.2
89
32
100
NS
357
5.0
203

50
NS
171
2.6
106

0
NS
15
0.1
16
42
100
1,920
NS
NS
NS

50
980
NS
NS
NS

0
12
NS
NS
NS
* Not sampled.
** Dosing system malfunctioned for 2 days spilling BRH sediments into all
treatments.
Chemical analysis of test sediments
88.	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 8).
Tissue residue
89.	Mytilus edutis. Differences in contaminant concentrations between
BRH and REF sediments facilitated the tracking of these contaminants in ex-
posed biota. Results of Experiment 1 indicated that PCB tissue residue con-
centration in mussels are directly related to exposure concentration
(Table 9). PCBs in mussels from the 0-percent BRH concentration remained
about the same over the 14-day experiment.
90.	The PCB tissue residue data from Experiment 2 are listed in
44

-------
Table 8
Concentrations of the Ten Selected Contaminants and Two Summary
Statistics for Both BRH and REF Sediments
Mean ฑ Standard Deviation
Sediment*
Chemical Compound
Phenanthrene
Sum of 178 alkyl
homologs
Fluoranthene
Benzo(a)pyrene
Ethylan
PCB as A1254
SUM of PAHs
CENT of PAHs
Copper
Cadmium
Chromium
Iron
BRH
REF
5,000 ฑ 1,800 (15)**
28,000 ฑ 8,300 (15)
6,300
3,900
4,000
6,400
142,000
232.8
2,900
24
1,480
31,000
ฑ 1,300 (15)
ฑ 970 (15)
ฑ 820 (15)
ฑ 840 (15)
ฑ 30,000 (15)
ฑ 1.7 (15)
ฑ 310 (18)
ฑ 1.0 (18)
ฑ 104 (18)
ฑ 2,800 (18)
85 ฑ 17 (12)
170 ฑ 26 (12)
240 ฑ 33 (12)
250 ฑ 28 (12)
0 ฑ - (12)
39 ฑ 4 (12)
4,500 ฑ 510 (12)
249.2 ฑ 1.7 (12)
60 ฑ 3 (15)
0.23 ฑ 0.04 (15)
50 ฑ 15 (15)
21,000 ฑ 1,400 (15)
* 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 CENT.
** (N) - number of replicates.
Table 10 and graphically depicted in Figure 11. Tissue residues, measured at
7-day intervals, Indicated that the mussels in the 0-percent BRH chamber main-
tained a relatively constant background concentration of PCBs throughout the
experiment. In the 10- and 30-percent BRH chambers, the concentration of PCBs
in the mussels increased from days 0 to 14 and then remained nearly constant
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 mus-
sels in the 30-percent BRH treatment was almost double that of mussels from
45

-------
Table 9
PCB Tissue Residues (ng/g Dry Weight) in Mussels from
Laboratory Experiment 1

% BRH

Day
0 50
100
0
117 117
117
14
154 2,100
3,700

Table 10

PCB
Tissue Residues (ng/g Dry Weight) in
Mussels from
Laboratory Experiment 2

% BRH

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
the 10-percent BRH treatment. The actual concentration of BRH dosed to the
30-percent BRH treatment, 3.3 mg/ฃ,, is nearly double that dosed to the
10-percent BRH treatment, 1.5 mg/j,. The measured whole water concentrations
of PCB were 11.86 and 23.57 ng/Jl for the 10- and 30-percent BRH treatments,
respectively. These data indicate a good relationship between the actual
dosed concentrations of BRH suspended sediment, the measured whole water con-
centrations, and the PCB tissue residues in the mussels in Experiment 2.
91. A comparison of the tissue residues between the two experiments can
be made for days 0 and 14. The PCB residue concentrations in the day 0 mus-
sels from Experiment 1 were almost half that of those from Experiment 2, (117
and 210 ng/g, respectively). In addition, day 14 PCB residue 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. PCB data from these experiments
were normalized to nanograms per gram of lipid, and the results are presented
46

-------
4000n
3500-
• TIME ZERO
O0% BRH
~ 10 % BRH
A 30% BRH
T3
ป 3000-
9
2500-
m
2000-
ซ/J
1500-
1000-
500-
O-
21
28
0
14
7
SAMPLING DAY
Figure 11. Concentrations of PCB as A1254 in the
tissue of M. edulis exposed to BRH suspended sed-
iments for 28 days
in Table 11 and Figure 12. Inspection of these data show that differences be-
tween experiments 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 12).
Table 11
PCB Concentrations (ng/g Lipid) in Mussels
from Both Laboratory Experiments
% BRH Treatments
Day
0*
0**
10**
30**
50*
0
2,900
2,400
2,400
2,400
2,900
7
—
5,200
17,100
24,000
—
14
3,800
4,300
27,000
54,000
53,000
21
—
5,000
35,000
67,000
—
28
—
3,800
30,000
66,000
—
100*
2,900
119,000
* Experiment 1.
** Experiment 2.
47

-------
140000-1
S 60000-
™ 80000-
a
120000-
100000-
40000-
20000-
0
0 10 20 30 40 50 60 70 80 90 100
EXPOSURE CONCENTRATION (%BRH)
Figure 12. Concentrations of PCB as A1254, normalized
for lipids, in the tissue of M. edulis exposed to BRR
sediment for 14 days
92.	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 13-18.
93.	While each of these graphs will not be discussed at length, it is
interesting to note the relationship between the molecular weight of the or-
ganic compounds and tissue residue over time. The benzo(a)pyrene tissue resi-
dues 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 with the 10-percent BRH
treatment. Mussel residues for both of these treatments are elevated compared
with the 0-percent BRH treatments. 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 with the 0-percent BRH expo-
sure. These data would suggest that mussels have the ability to metabolize
and/or excrete the lower molecular weight PAHs, even during continuous
48

-------
• TIME ZERO
O0% BRH
~ 10% BRH
A 30% BRH
175-1
150-
125-
100-
75-
50-
cO
25-
21
0
7
14
26
SAMPLING DAY
a. Phenanthrene
4000-1
• TIME ZERO
O 0% BRH
~ 10% BRH
A 30% BRH
3500-
V)
o 3000-
2500-
> 2000-
ฎ 1500-
o 1000-
500-
0
7
14
28
SAMPLING DAY
b. Sum of 178 alkyl homologs
Figure 13. Concentrations of phenanthrene and sum
of 178 alkyl homologs in the tissue of M. edulis
exposed to BRH suspended sediments for 28 days
49

-------
800-1
• TIME ZERO
0 0% BRH
~ 10% BRH
A 30% BRH
700-
o 600-
500-
I- 400-
O 300-
200-
100-
0
7
14
21
28
SAMPLING DAY
a. Fluoranthene
• TIME ZERO
00% BRH
~ 10% BRH
A 30% BRH
175-1
150-
9
c 125-
z
ฃ ioo-
75-
N
ffi 50-
25-
of
0
28
21
14
7
SAMPLING OAY
b. Benzo(a)pyrene
Figure 14. Concentrations of fluoranthene and
benzo(a)pyrene in the tissue of M. edulis ex-
posed to BRH suspended sediments for 28 days
50

-------
14 000
12000-
>ป
I.
"O
10000-
- 8000-
o 6000-
2
3
in
4000-
2000-
230n
~ 225-j
o
E
220-
<
a.
ฃ 2'5H
o
H 210-j
z
bJ
O
205-
200
• TIME ZERO
O 0% BRH
~ 10% BRH
A 30% BRH
a.
SAMPLING DAY
SUM of PAHs
• TIME ZERO
O 0% BRH
~ 10% BRH
A 30% BRH
~r
7
I
14
T-
21
—I
28
SAMPLING DAY
b. CENT of PAHs
Figure 15. Concentrations of SUM of PAHs and CENT
of PAHs in the tissue of M. edulie exposed to BRH
suspended sediments for 28 days
51

-------
1000
900-
800-
10 700-
c 600-
ซJ 500-
• TIME ZERO
O 0% BRH
~ 10% BRH
A 30 % BRH
i= 400-
300-
200-
100-
26
SAMPLING DAY
a. Ethylan
4000-1
3500-
• TIME ZERO
0 0% BRH
~ 10 % BRH
A 30% BRH
e 3000-
9
* 2500-
m
2000-
(O
o 1500-
1000-
500-
7
14
0
21
28
SAMPLING DAY
b. PCB as A1254
Figure 16. Concentrations of ethylan and PCB as
A1254 in the tissue of M, edulie exposed to BRH
suspended sediments for 28 days
52

-------
• TIME ZERO
O 0% BRH
~ 10% BRH
A 30% BRH
~1	1	
7	14
SAMPLING DAY
a. Cadmium
21
i
26
• TIME ZERO
O 0 % BRH
~ 10% BRH
A 30% BRH
SAMPLING DAY
b. Copper
Figure 17. Concentrations of cadmium and copper in
the tissue of M. edulis exposed to BRH suspended
sediments for 28 days
53

-------
8-1
• TIME ZERO
O 0% BRH
~ 10% BRH
A 30% BRH
7-
>>
W
TJ
o
7
14
21
28
SAMPLING DAY
a. Chromium
• TIME ZERO
0 0% BRH
~ 10% BRH
A 30% BRH
600-
450-
300-
150-
7
14
21
0
26
SAMPLING DAY
b. Iron
Figure 18. Concentrations of chromium and iron
in the tissue of M. edulis exposed to BRH sus-
pended sediments for 28 days
54

-------
exposure. In addition, the data would indicate that only higher molecular
weight compounds should be used to relate exposure levels and subsequent tis-
sue residue levels, because even under relatively constant exposure condi-
tions, residues of lower molecular weight PAHs did not reflect exposure
concentrations.
94.	Nephtys inaisa. Nephtys inoisa tissues from suspended sediment
laboratory exposures were analyzed for a suite of organic and inorganic con-
taminants found in BRH sediment. These tissue residues were measured on sam-
ples from days 0, 28, and 42 of the experiment. The summary statistics, SUM
and CENT, of the PAHs were also calculated for each of these sampling dates.
The tissue residue data for the representative subset of chemical compounds
are presented graphically in Figures 19-24.
95.	Although the data presented in these figures are not discussed in
detail (see Lake et al. 1987), some general observations are made. The tissue
residue concentrations of all the organic compounds increased with increasing
exposure. The PAHs, with the exception of fluoranthene, reached their highest
measured tissue concentrations at day 28 and exposure concentrations of
100-percent BRH (200-mg BRH/i.) • By day 42, the residue concentrations of
phenanthrene and benzo(a)pyrene declined by 30 and 50 percent, respectively.
The tissue residue concentrations of PCBs reached an apparent steady-state at
the 50-percent BRH (100 mg/BRH/fc) exposure by day 28, although there was a
continued increase at 100-percent BRH (200 mg/BRH/&) at day 42. Because of
its kinetic, partitioning, and persistence properties, PCB was selected as a
"tracer" for BRH material and was used to relate BRH exposure conditions to
tissue concentrations. Copper and cadmium, which have soluble fractions in
seawater, did produce elevated tissue concentrations as a consequence of in-
creased exposure to BRH suspended sediment. Chromium and iron, which are
bound to particulates, did not produce elevated tissue concentrations and in
fact showed apparent depuration of these compounds from day 28 to day 42 of
the experiment.
Effects results
96.	Mutilue edulia. Adenine nucleotide concentrations were measured in
adductor muscle tissues of M. edulie exposed to BRH sediments in both labora-
tory experiments. The results of these measurements are presented in
Table 12. These data are presented graphically in Figure 25. The data for
day 14 from both experiments represent an exposure range of 0 to 10 mg BRH/ฃ.
55

-------
TIME ZERO
0% BRH
50% BRH
100% BRH
10-
9-
tr
6-
5-
3-
0
14
28
42
DAY
a. Phenarithrene
CO
o
o
_i
o
2
0
1
>-
CO
N
2
a
CO
TIME ZERO
1400
300-
1200-
1000
900-
800-
700-
600-
500-
400-
300-
DAY
b. Sum of 178 alkyl homologs
Figure 19. Concentrations of phenanthrene and sum
of 178 alkyl homologs in the tissue of N. incisa
exposed to BRH suspended sediments for 42 days
56

-------
330-
• TIME ZERO
O 0% BRH
~ 50% BRH
A 100% BRH
* 300-
c 270-
180-
50-
O 120-
90-
60-
30-
0
14
28
42
DAY
a. Fluoranthene
0-
TIME ZERO
0% BRH
50% BRH
100% BRH
100-
90-
o>
70-
Lul
20-
0-
0
42
2B
14
DAY
b. Benzo(a)pyrene
Figure 20. Concentrations of fluoranthene and
benzo(a)pyrene in the tissue of N. inoisa ex-
posed to BRH suspended sediments for 42 days
57

-------
5500-
• TIME ZERO
O 0 % BRH
~ 50% BRH
A 100% BRH
5000-
3500-
* 3000-
CL
2500-
2000-
= 1500-
1000-
500-
0
14
28
42
DAY
a. SUM of PAHs
230-
227-
226-
225-
• TIME ZERO
O 0% BRH
~ 50% BRH
A 100% BRH
224-
o 223-
222-
221-
220-
219-
42
28
14
b. CENT of PAHs
Figure 21. Concentrations of SUM of PAHs and CENT
of PAHs In the tissue of N. inciaa exposed to BRH
suspended sediments for 42 days
58

-------
TIME ZERO
0% BRH
50% BRH
1007. BRH
100-
90-
80-
o>
70-
60-
50-
40-
30-
20-
10-
0*
0
14
28
42
DAY
a. Ethylan
• TIME ZERO
O 0% BRH
~ 50% BRH
A 100% BRH
DAY
b. PCB as A1254
Figure 22. Concentrations of ethylan and PCB as
A1254 in the tissue of N. inciea exposed to BRH
suspended sediments for 42 days
59

-------
TIME ZERO
0% BRH
50% BRH
100% BRH
>N
o>
0 7-
06-
05
O
14
28
42
DAY
a. Cadmium
1100-
TIME ZERO
0 % BRH
50% BRH
100% BRH
1000-
900-
T>
800-
700-
600-
a 500-
o 400-
300-
200-
100-
42
28
14
0
DAY
b. Copper
Figure 23. Concentrations of cadmium and copper in
the tissue of N. ineisa exposed to BRH suspended
sediments for 42 days
60

-------
50-
45-
40-
35-
30-
25-
• TIME ZERO
O 0% BRH
~ 50% BRH
A 1007. BRH
20-
10-
5-
0
14
28
42
0 AY
a. Chromium
• TIME ZERO
O 0% BRH
~ 50% BRH
A 100% BRH
—I
42
T-
14
-T"
28
DAY
b. Iron
Figure 24. Concentrations of chromium and iron
in the tissue of N. inoisa exposed to BRH sus-
pended sediments for 42 days
61

-------
Table 12
Adenine Nucleotide Concentrations (yimol/g Wet Weight) in
M. edulis from the Two Laboratory Experiments
Treatment
% BRH
ATP
ADP


AMP
Total
AEC


Experiment
1,
Day
14


0
5.01
1.23


0.07
6.31
0.89
50
3.83
1.19


0.13
5.15
0.86
100
3.68
0.83


0.05
4.56
0.90


Experiment
2,
Day
14


0
4.26
1.64


0.38
6.28
0.81
10
3.29
1.76


0.60
5.65
0.73
30
3.09
1.80


0.65
5.55
0.71


Experiment
2,
Day
28


0
3.93
1.41


0.22
5.56
0.83
10
4.07
1.40


0.25
5.71
0.83
30
3.90
1.61


0.35
5.86
0.80
These data were pooled and analyzed for correlations between BRH exposure con-
centrations and adenine nucleotide concentrations. The only significant rela-
tionship occurred between BRH exposure and the adenine nucleotide pool
2
(P = 0.001, r = 0.94). The other adenine variables and the AEC showed no
significant relationship with BRH exposure concentrations.
97.	Nephtys inaisa. Adenine nucleotide concentrations were measured in
whole worms exposed to BRH sediments in three laboratory experiments. The re-
sults of these measurements are presented in Table 13. These data are pre-
sented graphically in Figure 26. The data for day 28 from the second and
third experiments were pooled and analyzed for correlations between BRH expo-
sure concentrations and adenine nucleotide responses. There were no signifi-
cant relationships between BRH exposure and adenine nucleotide pool concentra-
tions, other adenine variables, or AEC.
Field
Exposure
98.	Mytilus edulis exposures estimated from tissue residues. The first
method used to estimate exposure conditions of M. edulis to BRH material in
62

-------
7 -|
6 -
5-
r 4-
v
i
o
ฃ
(/)
_j
O
O
Q.
liJ
Q
_)
U
3
Z
2
UJ
o

-------
Table 13
Adenine Nucleotide Concentrations (pmol/g Wet Weight) In
N. ineisa from the Three Laboratory Experiments
Treatment
% BRH
ATP
ADP


AMP
Total
AEC


Experiment
1,
Day
10


0
1.55
0.53


0.12
2.20
0.82
50
1.83
0.40


0.05
2.28
0.88
100
1.53
0.53


0.06
2.12
0.83


Experiment
2,
Day
28


0
0.75
0.34


0.08
1.17
0.78
50
1.63
0.60


0.11
2.35
0.81
100
1.93
0.55


0.08
2.56
0.83


Experiment
3,
Day
28


0
1.65
0.65


0.06
2.37
0.82
50
1.52
0.50


0.09
2.11
0.83
100
1.04
0.47


0.07
1.59
0.79


Experiment
3,
Day
42


0
1.74
0.54


0.08
2.35
0.85
50
1.28
0.46


0.10
1.84
0.81
100
1.68
0.49


0.07
2.24
0.84
demonstrate several spatial and temporal trends (Table 14). Spatially, the
data indicate a trend towards greater exposure near the CNTR station immedi-
ately following disposal. This is evidence by the elevated exposures at T = 0
(1000E > 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.
100. 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!% 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, de-
creased to between 0.7 and 0.3 mg/fc, half that of the previous collection.
64

-------
EXP 1 : 10 DAY
0.5 -
0.0 -
EXP 2 - 28 DAY
EXP 3 28 DAY
lu 2.0 -
—
TOTAL



	•
•o		
ATP
	-0
•o
AEC



— Q

AOP
	ฆ
-ฆ—
•A—
AMP
	A
O
3
z
5 -
Ul
5 l.O
2 0.5 -I
EXP 3 42 DAY
BRH (mq /L)
200
Figure 26. Response of adenine nucleotide
pools in N. incisa to BRH exposure in
laboratory experiments
Subsequent collections indicated a continued decrease to levels similar to
those at the REFS station by T + 12.
101. Exposures estimated from water chemistry data. In addition to the
estimates of BRH exposure based on mussel PCB tissue residues, a second
65

-------
Table 14
Predicted BRH Suspended Material Sediment Exposure (mg/I)
Required To Achieve the Measured Tissue Residue
Values of Mussels Deployed in CLIS*
Collection

Estimated
Exposure Range
Cruise
Station
High Value
Low Value
H
1
O
CNTR
0.37
0.00

400E
0.26
0.00

1000E
0.38
0.00

REFS
0.38
0.00
T = 0
1000E
1.04
0.56

REFS
0.49

T + 2
400E
1.39
0.79

1000E
0.98
0.38

REFS
0.60

T + 8
CNTR
0.67
0.21

400E
0.71
0.25

1000E
0.60
0.14

REFS
0.46

T + 12
CNTR
0.61
0.06

400E
0.64
0.09

1000E
0.53
0.00

REFS
0.55

T + 15
CNTR
0.84
0.31

400E
0.61
0.08

1000E
0.53

T + 21
CNTR
0.52
0.12

400E
0.66
0.26

1000E
0.55
0.15

REFS
0.40

T + 27
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.
66

-------
Table 14 (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 CNTR	0.52	0.00
400E	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
67

-------
estimate 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 15).
102.	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 com-
pared with the REFS station. The same pattern was observed in both the copper
and PCB estimates for the 21 July 1983 sample. A decreasing concentration of
BRH material was estimated moving away from the CNTR station.
103.	On a temporal scale, the BRH concentration (copper data) decreased
by about half from June to July (high estimate). After this collection, how-
ever, the copper-based BRH estimates fluctuated over time with the December
1983 and June 1984 values higher than the September 1983 concentrations. This
pattern over time may be more reflective of CLIS than of actual BRH levels be-
cause these estimates (high value) were based on the absolute copper levels at
each location. Inspection of the low estimate indicated a more distinct pat-
tern 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.
104.	The pattern of BRH 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 in-
creasing time. In addition, the high estimates did not show the same varia-
bility over time that the copper data did. This may indicate that PCB concen-
trations in Long Island Sound were most constant over time and thus BRH
estimates based on PCB concentrations were less influenced by background fluc-
tuations .
105.	Nephtys inaisa exposure estimated from tissue residues. The first
method used to estimate exposure conditions of N. inaisa to BRH material in
CLIS involved the laboratory-generated relationships between PCB tissue resi-
dues and BRH exposures. Using this relationship and the PCB tissue residues
in field-collected N. inaisa, estimates of field BRH exposure concentrations
68

-------



Table 15



Predicted BRH
Suspended
Sediment Exposure
(mg/I) Based
on

PCB and
Copper Whole Water Chemistry Data*







Collection

Estimate Using Copper
Estimate
Using PCB
Cruise
Station
High Value Low Value
High Value
Low Value
T + 2
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
T + 9
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
T + 15
CNTR
	
	
0.17
0.07

400E
—
—
0.21
0.11

1000E
—
—
0.16
0.06

REFS
—
—
0.10
0.00
T + 15
CNTR
	
—
—
—

400E
0.72
0.22
—
—

1000E
—
—
—
—

REFS
0.50
0.00
—
—
T + 28
CNTR
	
	
0.05
0.00

400E
1.13
0.37
0.08
0.00

1000E
—
—
0.09
0.00

REFS
0.91
0.00
0.09
0.00
T + 55
CNTR
	
__
	
—

400E
1.00
0.09
—
—

1000E
—
—
—
—

REFS
0.91
0.00
—
—
* Each estimate was calculated through division of the concentration of PCB
or copper present in the field by the concentration of that material present
in the BRH barrel material (6,910 ng/g and 2,900 yg/g for PCB and copper,
respectively). The high value was determined from the actual whole water
concentration while the low value was calculated after the REFS values were
subtracted.
69

-------
were calculated. There are several assumptions in this approach: N. inaisa
provides an integrated measure of exposure; N. inaisa tissue residues were at
steady-state with BRH exposure concentrations at the time of sampling; and
PCBs are a good chemical marker for BRH sediments. Based on the results of
the laboratory experiment, each of these assumptions seems reasonable.
106. The estimated exposures resulting from this approach are presented
as milligrams per litre BRH for each station and collection date in Table 16.
Table 16
Estimated Concentrations of BRH Sediment (mg/ft) Suspended at
Sediment-Water Interface Based on PCB Concentrations
in Field-Collected N. inoisa
	Station	
400E	1000E	REFS
0	—	0
0	—	2
9	—	3
15	—	8
95	43	2
114	44	2
131	88	12
51	26	0
38	10	0
29	10	3
5	4	0
Date

CNTR
17 Apr
82
—
16 Nov
82
—
16 Feb
83
—
12 Apr
83
—
02 Jun
83
—
03 Jul
83
—
06 Sep
83
—
29 Nov
83
—
20 Mar
84
47
16 Oct
84
53
24 Jan
86
76
Nephtys inaisa at CNTR were buried during disposal of the dredged material and
did not recolonize the dredged material mound until the spring of 1984. When
the worms recolonized the mound, sampling began. The data in Table 16 display
clear spatial and temporal trends. The highest estimates were in the immedi-
ate vicinity of the disposed BRH material (400E) during the summer of 1983.
There was a decrease in exposure at 400E and 1000E in 1984 and 1985.
107. Nephtys inaisa exposure estimates from physical data. Benthic ex-
posure at the FVP disposal site can occur through both the suspended and
bedded sediments. This section describes predictions of the maximum upper
70

-------
bound suspended sediment concentrations from 1 m above the bottom to the
sediment-water interface. This calculation is based upon the assumption that
the sediment-water interface consists totally of BRH sediment and that the
suspended solids at the sediment-water interface consist totally of BRH sedi-
ment and that the contaminant concentrations are similar to those found in the
dredged material prior to disposal.
108.	TSS concentrations were measured at the FVP site at 1 m above the
sediment-water interface with an in situ monitoring platform (Bohlen and
Winnick 1986). Although there is considerable variation in the data through
one tidal cycle, average background suspended solids were estimated to be
10 mg/ฃ, while during typical storm conditions suspended solids concentrations
averaged 30 mg/& for periods of 4 to 7 days (Munns et al. 1986).
109.	Using an acoustic profilometer, the suspended sediment concentra-
tions at 1 m above the bottom were found to increase exponentially to the sed-
iment-water interface. The upper and lower limits for this increase are 10x
and lx, respectively, depending on hydrographic conditions (Bohlen and Winnick
1986). These data, in conjunction with suspended sediment data for 1 m above
the bottom, can be used to predict the suspended solids concentrations at the
sediment-water interface.
110.	For example, the suspended solids concentration under background
conditions (10 mg/I) would increase to 100 mg/i for the 10x enrichment at the
sediment-water interface and decrease to 10 mg/i for the quiescent conditions.
Likewise, under storm conditions (30 mg/Jl), the sediment-water interface sus-
pended solids concentrations would range from 300 to 30 mg/i for the 10x and
lx enrichments, respectively (Figure 27). These conditions represent the max-
imum upper bound exposures that would be expected to occur at the sediment-
water interface and could be made using predisposal, site characterization
data.
111.	A more probable estimate is provided by using contaminant concen-
trations present in the sediment after disposal. It is this material that
will be resuspended and transported as suspended solids to populations outside
the disposal site. Assuming that resuspended surficial sediments are the
source of contaminants for the suspended sediments, the maximum upper bound
estimates can be adjusted to reflect the spatial and temporal changes in con-
taminant concentration. These changes are represented as percentages of BRH
sediment in the 0- to 2-cm surface layer at CNTR, 200E, 400E, and 1000E from
71

-------
1.0-1
Suspended solids (mg BRH/I)
at I m above bottom
A = 10
B = 30
tr 0.4-
u 0.3-
-i	1	1	r
0 20 40 60 80
00 120 A
i	i i i i i i
0 60 120 180 240 300 360 B
SUSPENDED SOLIDS (mg BRH/I)
Figure 27. Suspended sediment
concentrations from 1 in above
the bottom to the sediment-
water interface for storm and
background conditions
June 1983, immediately after disposal, to October 1985 (Table 17). The com-
bination of these percentages and the TSS concentrations predicted for the
sediment-water interface results in concentrations of BRH suspended sediments
at the sediment-water interface for each station and sampling date (Table 18).
112.	The sediment samples used for the percent calculations were not
replicated; therefore, no variability estimates are available. However, cer-
tain trends in the data are evident (Table 17). The percentages of BRH sedi-
ment (<50 percent) at CNTR and 200E were low compared with the barrel sedi-
ments collected predisposal. There is a gradient of BRH material that is a
function of both distance from the center of the mound and of time from dis-
posal. BRH sediment concentrations were highest at CNTR and 200E immediately
after disposal and decreased significantly through October 1984. Concentra-
tions were elevated in December 1984 at CNTR and 200E and again in October
1985 at 200E. The BRH concentrations at 400E also decreased through time and,
after December 1983, were the same or higher than those at 1000E.
113.	The 1- to 2-percent BRH sediment calculated for 1000E represents a
quantitatively measured elevation above background and is supported by tissue
residue data for N. inaisa. This contamination could have resulted from the
72

-------
Table 17
Percent BRH Sediment in the Surficial Sediments at the FVP Disposal Site
Station
Date
CNTR
200E
400E
1000E
Jun 83
44.5
41.1
12.5
1.8
Jul 83
15.0
37.4
3.3
1.6
Sep 83
32.0
36.7
4.9
2.0
Dec 83
32.8
36.1
9.5
4.4
Mar 84
4.4
2.2
1.9
1.8
Jun 84
9.5
15.6
0.5
0.7
Sep 84
10.0
0.8
3.5
0.5
Oct 84
2.6
—
0.2
1.6
Dec 84
35.1
11.3
0.0
1.0
Oct 85
0.2
21.0
0.0
0.0
Table 18
Concentration of BRH (mg/ft) at the Sediment-Water Interface for TSS
Concentrations of 30 mg/ft and 10 mg/& and an Enrichment of 10**




Station





CNTR

200E

400E

1000E
Date
300
100
300
100
300
100
300
100
Jun 83
133.5
44.5
123.3
41.1
37.5
12.5
5.4
1.8
Jul 83
45.0
15.0
112.2
37.4
9.9
3.3
4.8
1.6
Sep 83
96.0
32.0
110.1
36.7
14.7
4.9
6.0
2.0
Dec 83
98.4
32.8
108.3
36.1
28.5
9.5
13.2
4.4
Mar 84
14.2
4.4
6.6
2.2
4.7
1.9
5.4
1.8
Jun 84
28.5
9.5
46.8
15.6
1.5
0.5
2.1
0.7
Sep 84
30.0
10.0
2.4
0.8
10.5
3.5
1.5
0.5
Oct 84
7.8
2.6
—
—
0.6
0.2
4.8
1.6
Dec 84
105.3
35.1
33.9
11.3
0.0
0.0
3.0
1.0
Oct 85
0.6
0.2
63.0
21.0
0.0
0.0
0.0
0.0
* BRH concentrations for the 1* enrichment can be obtained by dividing the
tabular values by 10.
73

-------
dispersion of dredged material during disposal, the errant disposal of BRH
material in the vicinity of 1000E, or the continuous transport of contaminated
material from the disposal mound.
114.	The estimates of exposure to BRH material at the sediment-water
interface derived from tissue concentrations of PCB and from the maximum upper
bound predictions agreed well. The exposure estimates based on the chemistry
of the 0- to 2-cm surface sediments were low. If the exposure estimates based
on tissue concentrations of PCB are accepted as a valid check on the exposure
estimates from the physical data, it is concluded that the higher estimates of
exposure are accurate. The simplest explanation is that the 0- to 2-cm sam-
pling procedure integrates clean and contaminated sediments, thus underesti-
mating the actual exposures experienced by the worms. The data suggest that
the worms were exposed to a thin, surface layer of contaminated sediment.
Tissue residues
115.	Mytilus edulis. The tissue residue levels for the mussels col-
lected during the course of the FVP study are presented graphically in Fig-
ures 28-33 for each of the 12 selected organic, inorganic, and summary statis-
tic chemical contaminants. The raw data shown on these figures are included
in Appendix B.
116.	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.
117.	A consistent pattern emerged when the spatial component of the
organic residue data was considered within a sampling date. Mytilus edulis
were deployed during the actual disposal operation, and were collected 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 REFS. Once again, a con-
sistent pattern 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 tissue residue
concentrations between stations decreased dramatically.
118.	The tissue residue data for metals did not provide as clear a pic~
ture of the disposal operation as the organic residues. In general, metal
74

-------
ฆ	CNTR
ฆ	400E
~ IOOOE
Qrefs
jJk

4/83 5/83 6/83 7/83 8/83 9/83 10/83 11/83 3/84 6/84 8/85
SAMPLING DATE
~r
t	r
t	r
"T~
t	r
T-4 T + 0 T+2 T+8 T+12 T+15 T+21 T+27 T+43 T+55 T+II6
CRUISE NUMBER
1
a. Phenanthrene
2000
O
oป
(A
ID
O
J
O
2
O
X
>-
*
_I
<
GO
N
U_
O
2
3
(/)
1500
1000
500
ฆ	CNTR
ฆ	400E
~	IOOOE
~	REFS
,BTi t	| Iti,
r
4/83 5/83 6/83 7/83 8/83 9/83 10/83 If/83 3/84 6/84 8/85
SAMPLING DATE
I	1	I	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
b. Sum of 178 alkyl homologs
Figure 28. Concentrations of phenanthrene and the sum of 178 alkyl
homologs in the tissues of M. edulis exposed at the specified FVP
stations and sampling dates
75

-------
ฆ	CNTR
ฆ	400E
~	IOOOE
~	refs
i	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
T-4 ' T+0 ' T+2 ' T+8 ' T + 12 ' T+15 ' T+21 ' T+27 ' T+43 ' T+55 ' T+U6 '
CRUISE NUMBER
a. Fluoranthene
250-1
200-
I 50
100
50-
| CNTR
|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
I	1	I	1	1	1	1	1	1	1	1	—i
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. Benzo(a)pyrene
Figure 29. Concentrations of fluoranthene and benzo(a)pyrene
in the tissues of M. edulis exposed at the specified FVP
stations and sampling dates
76

-------
9000-1
? 6000
I
<
a.
2
=>
(/>
3000
| CNTR
ฆ 400E
~	I000E
~	refs
th y y fci irh lii - n, rh,
T/83 5/83 ' 6/83 ' 7/83 ' 8/83 ' 9/83 ' 10/83' 11/83' 3/84 6/84 8/8S
SAMPLING DATE
t	r
-i	1	1	—r
T

T-4 T+0 T+2 T+8 T+12 T+15 T+21 T+27 T+43 T455 T+II6
CRUISE NUMBER
a. SUM of PAHs
250-1
240
x
2
o
tc
I-
z
UJ
o
CNTR
400E
~ I000E
230
220
210 ™ i	i ~ |"	|	|	i	|	i	i
4/83 5/83 6/63 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
b. CENT of PAHs
Figure 30. Concentrations of the SUM of PAHs and CENT of PAHs
in the tissues of M. edulie exposed at the specified FVP
stations and sampling dates
77

-------
CNTR
ฆ 400E
~ IOOOE
LJREFS
4/83 6/63 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+l 16 '
CRUISE NUMBER
a. PCB as A1254
120
100-
80-
< 60H
-J
>*
X
ui 40 H
20
.mฃ1
CNTR
|400E
~ IOOOE
4/83 5/83 6/83 7/83 8/83 9/83 10/83 II/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+-I5 T+21 T+27 T+43 T+55 T + II6
CRUISE NUMBER
b. Ethylan
Figure 31. Concentrations of PCB as A1254 and ethylan in
the tissues of M. edulis exposed at the specified FVP
stations and sampling dates
78

-------
6 -|
ฆ	CNTR
ฆ	400E
~	IOOOE
~	refs
4/83 S/83 6/83
7/83 8/83 9/83 10/83 11/83
3/84 6/84 8/8S
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
a. Cadmium
10 20
o 10
| CNTR
|400E
0 IOOOE
~ refs
i —r
4/83 S/83 6/83 7/83 8/83 9/83 10/83 II/83 3/84 6/84 8/8S
SAMPLING OATE
T-4 ' T+0 ' T+2 ' T+8 ' T+I2 ' T+I5 ' T+2I ' T+27 ' T+43 ' T+55 ' T+II6 '
CRUISE NUMBER
b. Copper
Figure 32. Concentrations of cadmium and copper in
the tissues of M. edulis exposed at the specified FVP
stations and sampling dates
79

-------
4-i
|CNTR
ฆ 400E
~	IOOOE
~	REFS
4/83 5/83 6/83
7/83 8/83 9/83 10/83
1/83 3/84 6/84 8/85
SAMPLING DATE
I	I	I	1	I	I	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
>.
w
"O
1000 -
z
o
at
500-
ฆ	CNTR
ฆ	400E
~	IOOOE
~	REFS
T
H
4/83 5/83 6/83 7/63 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
b. Iron
Figure 33. Concentrations of chromium and iron in
the tissues of M. edulis exposed at the specified FVP
stations and sampling dates
80

-------
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). The October and November sam-
ples consisted of organisms that had been deployed at the FVP site for 7 and
3 months, respectively. One possible explanation for the difference between
elevated metal and organic tissue residue patterns may be that mussels require
a long period of time to reach steady-state with respect to metal concentra-
tions. Comparing the organic and metal residue data from the field suggests
that organic tissue residues present a better picture of the disposal opera-
tion at the FVP disposal site.
119.	Nephtys inaisa. The tissue concentrations for the N. incisa col-
lected at the CLIS site during the FVP study are presented graphically for
each of the 12 selected chemical variables in Figures 34-39. The raw data
shown on these figures are included in Appendix B.
120.	Clear spatial and temporal patterns of tissue concentrations of
PCBs and PAHs were found. Highest tissue concentrations were determined at
station 400E with lowest concentrations at station REFs. When N. inaisa re-
colonized the dredged material site at station CNTR in the spring of 1984, the
tissue concentrations of PCBs in these worms were comparable with those found
at 400E immediately postdisposal.
121.	The temporal patterns of the field tissue residue show a rapid in-
crease in organic residue values during and immediately postdisposal at 400E
and at 1000E. The PAH residues for N. inaisa showed an increase immediately
postdisposal. This was followed by a rapid decline during July and August.
The phenanthrene residue value returned to background levels by September, but
the higher molecular weight PAH tissue residues tended to remain at approxi-
mately 25 percent of their maximum values for an additional year. The PCB
residues at 400E increased rapidly immediately after disposal and, unlike the
PAHs, remained elevated through September and declined only 50 percent by
December 1983. Unlike the PAHs, PCB residues increased 2.5 times REFS at
1000E postdisposal and remained elevation above REFS until October 1984.
There were no clear temporal or spatial patterns for inorganic tissue residues
for N. inaisa from the field.
81

-------
600
O CNTR
• 400E
~ I000E
REPS
400
q: 300
i rff i f i i
I t I I I "rf'f i Pi i ฅ i i	i i i i i I i i!
asondjfmamjjasondjfmamjjasondjfmamjjasondj
1982
1983
1984
1985
T+0 T+16 T+28 T+44
T t+74
T ~ 140
a. Phenanthrene
6000
? 5000
4000
3000
2000-
I 000
o CNTR
400E
~ IOOOE
1 'nฎrf 1* i f i i P"m<-i i i i i iA
'I I I I I I I I I I II
ASONDJFMAMJJASONDJFMAMJJASONDJFMAMJJASONDJ
1982
1983
1984
1985
TปD T +16 T +28 T+44
T + 140
b. Sum of 178 alkyl homologs
Figure 34. Concentrations of phenanthrene and the sum of 178 alkyl
homologs in the tissues of N. inaiaa collected at the specified FVP
stations and sampling dates
82

-------
1600
1400
1200
- 1000-
i 800
<
a.
o
3
600-
400-
200

O CNTR
400E
D lOOOE
lllllllllTlllllTllllllllllll
AS ONDJFMAMJ JASONDJ FMAMJJASON DJFMAMJJASONDJ
1982
1983
_l	I	L.
1984
1985
T +0 T +16 T +28 T +44
T+74
T + 140
a. Fluoranthene
500
* 400
a:
>
a
z
Ul
CD
300-
200-
100-
O CNTR
• 400E
0 lOOOE
ฆ REFS

'II "I I I II I I I II
M I M I I M M I II II I I I I II I I I I I I I I M I
ASONDJFMAMJJASONDJFMAMJJASONDJFMAMJJASONDJ
1982
1983
1984
1985
_i	l	u
T+0 T + 16 T+28 T+44	T+M
T+140
b. Benzo(a)pyrene
Figure 35. Concentrations of fluoranthene and benzo(a)pyrene
in the tissues of N. inaisa collected at the specified FVP
stations and sampling dates
83

-------
20 000-;
17 500
15000
12 500-
10000-
5
D
7 500
5000H
2 500
o CNTR
• 400E
~ I000E
REFS
i i i i i i i i i i i i i i i i i i n i i i i i i i ii i i i i i i
ASONOJFMAMJJiSONDJFMAMJJASONDJFMAMJJASONDJ
1982
1983
1984
1985
_J	L-
T+0 T+16 T + 28 T + 44	T + 74
T +140
a. SUM of PAHs
270 -i
260
* 250
o
E
O 2 30
o
o
IE
y- 2 20-j
z
Ul
o
2 10
200
o CNTR
• 400E
~ I000E
I I I I I i i I I I I I I I I I I I I I i i i I i i i I I i I i i I i I I i I I I i
ASONDJFMAMJJASONDJFMAMJJASONDJFMAMJJASONDJ
1982
1983
1984
1985
_l
T + 0 T+16 T +28 Tf44	T*74
b. CENT of PAHs
T+140
Figure 36. Concentrations of the SUM of PAHs and CENT of PAHs
in the tissues of N. inaisa collected at the specified FVP
stations and sampling dates
84

-------
1400-
1200
1000
ฃ 800-
V
N
< 600-

o
o.
400-
200-
o CNTR
400E
~ IOOOE
0 I I I I I 1 I 1 I I I I I I I II I I I I I I I I I I I I I i I I I I I I I I 1 i i I
ASONDJFMAMJJAS0NDJFMAMJJAS0NDJFMAMJJAS0NDJ
1982
1983
_i	I	u
1984
1985
Tซ0 Tt|6 T+28 T+44
Tซ74
T-M40
z
<
I 6-|
I 2
I 0-
8-
2-
a. PCB as A1254
O CNTR
• 400E
a ioooe
ฆ REFS
oiiiiiii^PiyfiPiitiiiiiiiiiii^iiiiiiiiiiiiiif
A SONDJFMAMJJAS OND JFMAMJJ ASONOJ FMAMJJ AS0NDJ
1982
1983
-I	1	L_
1984
1985
_L_
T+0 T+16 T+28 T+44
T+74
T + 140
b. Ethylan
Figure 37. Concentrations of PCB as A125A and ethylan in
the tissues of N. incisa collected at the specified FVP
stations and sampling dates. The value for CNTR on
March 1984 (Figure 37b) was equal to REFS
85

-------
O CNTR
v 200E
400E
~ IOOOE

111 i 11 11 i i 11 11 i 11 i 11 i i i 11 i 11 i 11 i 111 11 i
JFMAMJJASONDJFMAMJJASONDJFMAMJJASONDJF
1963

_i_
_L_
1984
_l_
1985
T + O T+16 T+28	T+44 T + 74
a. Cadmium
T +140
0.
a
o
o
250-,
200
150-
100
50
o CNTR
7 200E
• 400E
~ IOOOE
ฆ REFS
-V
-B
I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I I
JFMAMJJASONDJFMAMJJASONDJFMAMJJASONDJF
1983
1984
1985
T + 0 T+16 T+28
b.
T +44 T + 74
Copper
T + 140
Figure 38. Concentrations of cadmium and copper in
the tissues of N. inaisa collected at the specified
FVP stations and sampling dates
86

-------
12
I 0
s
=5
6-
2
o
tr
x
ฐ 4
O CNTR
r 200E
ฆ
0 I I I I I I I I I I I I I | I I I I i | | I I i I | I I | I I I I I I I I I I
JFMAMJJASONDJFMAMJJASONDJFMAMJJASONDJF
1983
1984
1985
_l_


_1_
T + O T+16 T + 28
T 444
T + 74
T + 140
O
a.
2000
1750
150.0
1250
1000
750-
500
250
a. Chromium
O CNTR
V 200E
• 400E
D IOOOE
i i i i i I I l I I I I I I I i i i—I I i i i i i i i i i i i i i i i i i i
JFMAMJ JASONOJ FMAMJ J AS ONDJ F MAMJ JASONDJF
1983
1984
1985

T+O
T+16 T + 28
T + 74
T + 140
b. Iron
Figure 39. Concentrations of chromium and iron in
the tissues of N. inoisa collected at the specified
FVP stations and sampling dates
87

-------
Effects results
122.	Mytilus edul'Ls. Adenine nucleotide concentrations were measured
in adductor muscle tissue of M. edulis exposed at the FVP field stations. The
results of these measurements are presented in Table 19. The data for adenine
nucleotide pool concentrations and for AEC are presented graphically in Fig-
ure 40. The physiology of M. edulis is influenced by a variety of seasonal
factors that varied naturally from collection to collection during the FVP.
Because seasonal influences might conceal responses to the BRH sediments, the
field exposure-effects data will be considered within discrete sampling peri-
ods, where mussels were presumably exposed to similar seasonal conditions. As
was stated at the beginning of this report, the purpose for using adenine
nucleotide responses was to determine whether relative sublethal effects could
be measured between laboratory treatments and field stations. Consideration
of field exposure-effects relationships within a sampling period is entirely
consistent with this objective.
123.	The use of field measurements of adenine nucleotide concentrations
within a sampling period reduced sample size to a maximum of four when all
stations were sampled. Regression analysis with this sample size is not ap-
propriate; therefore, the data are presented graphically to illustrate trends.
Because detailed field exposure data are not available, tissue concentrations
are used to represent exposure concentrations experienced by the organisms.
It would be impractical to present graphs for all of the residue-effects data
for each of the 11 sampling dates. PCBs were selected because of the good re-
lationship previously described between tissue concentrations of PCBs and BRH
exposure concentrations in laboratory experiments. Therefore, only the resi-
due data from the field for PCBs are used. These data are related to the ef-
fect responses of total adenylate nucleotide pool concentration and AEC. The
total adenylate nucleotide pool concentration was the only adenylate nucleo-
tide measurement that responded to BRH exposure in laboratory experiments.
The AEC is included here because it is the primary response being evaluated in
this report. When the data were examined in this manner, there were no trends
among stations on any sampling dates for total adenine nucleotide pool concen-
trations or for AEC.
124.	Nephtys inaisa. Adenine nucleotide concentrations were measured
in whole worms sampled at the FVP stations on the specified dates. The re-
sults of these measurements are presented in Table 20. The data for total
88

-------


Table
19


Adenine
Nucleotide Concentrations
(ymol/g Wet Weight)
i Measured in


Adductor
Muscle Tissues
of M. edulis Sampled at


FVP
Field Stations
on Specific Dates


Station
ATP
ADP
AMP
Total
AEC


22 Apr 83
(T - 04)


CNTR
3.00
0.83
0.07
3.90
0.88
400E
3.06
0.83
0.11
4.00
0.87
1000E
2.74
0.79
0.09
3.62
0.86
REFS
2.94
0.80
0.05
3.79
0.88


24 May 83
(T - 0)


CNTR
	*
	
__
_ _

400E
—
	
	
	
	
1000E
3.69
0.98
0.12
4.79
0.87
REFS
3.40
0.85
0.13
4.38
0.87


07 Jun 83
(T + 2)


CNTR

	
...
ซ...
_ _.
400E
3.65
0.82
0.05
4.52
0.90
1000E
3.96
1.09
0.14
5.19
0.87
REFS
—
—
—
—
—


13 Jul 83
(T + 8)


CNTR
3.30
1.32
0.23
4.85
0.82
400E
3.59
1.28
0.19
5.06
0.84
1000E
3.40
1.19
0.24
4.83
0.83
REFS
2.84
1.20
0.32
4.36
0.83


10 Aug 83
(T + 12)


CNTR
3.71
1.47
0.39
5.57
0.80
400E
3.79
1.18
0.26
5.23
0.84
1000E
3.47
1.24
0.35
5.06
0.81
REFS
3.87
1.84
0.95
6.66
0.83
(Continued)
* Not sampled.

-------
Table 19 (Concluded)
Station
CNTR
400E
1000E
REFS
CNTR
400E
1000E
REFS
CNTR
400E
1000E
REFS
CNTR
400E
1000E
REFS
CNTR
400E
lOOOE
REFS
CNTR
400E
lOOOE
REFS
ATP
2.72
2.61
2.79
2.31
	*
3.27
4.06
3.86
2.47
2.48
2.71
ADP
AMP
3.84
3.52
2.53
2.52
2.64
3.34
3.23
2.90
3.95
06 Sep 83 (T + 15)
1.33
1.54
1.49
1.22
0.42
0.50
0.43
0.42
29 Nov 83 (T + 27)
1.07
0.93
1.08
0.13
0.06
0.09
20 Mar 84 (T + 43)
0.88
0.73
0.06
0.04
0.94	0.08
05 Jun 84 (T + 55)
0.89	0.10
0.91	0.11
17 Oct 84 (T + 74)
1.05
1.00
1.10
0.15
0.14
0.15
14 Aug 85 (T + 116)
1.17
1.17
0.94
1.15
0.17
0.22
0.14
0.08
Total
4.47
4.65
4.71
3.95
4.47
5.05
5.03
3.41
3.25
3.73
4.83
4.54
3.73
3.66
3.89
4.68
4.62
3.98
5.18
AEC
0.76
0.73
0.75
0. 74
0.85
0.90
0.88
0.85
0.88
0.85
0.89
0.88
0.82
0.82
0.82
0.83
0.82
0.84
0.87
* Not sampled.
90

-------
i.oo-
0.95-
0.90-
0.85
0.80
0.75H
0.70
0% BRH
A
A
10% BRH
A
30% BRH
A

1.00-j

0.95-
(A

2
0.90-
E

>s
in
0.85-
c

01
a
0.80-
o


0.75-
a

LlI
<
0.70-
1.00-1
0.95
0.90
0.85-
0.80-
0.75-
0.70-
a. Laboratory
rP
s
b. T-4, 22 APR 83
O CNTR
A 400 E
~ I000E
<7 REFS
T
3000
5
-4
-3
4000
1000	2000
PCB AS AI254 (ng/g dry)
c. Tซ 0,24 MAY 83
Figure 40. Relationship between adenine nucleotide con-
centration and AEC in M. edulis and PCB tissue residue
concentrations in laboratory- and field-exposed animals.
The laboratory data are presented to provide a perspec-
tive between the residue concentrations of laboratory-
and field-exposed mussels (Sheet 1 of 4)
91

-------


O
CNTR

A
A
400E
~

~
I000E


V
REFS
4"
d. T + 2, 7 JUN 83
aI
	,	,	r
e. T + 8, 13 JUL 83
f
ฆI

-4
4000
1000	2000	3000
PCB AS AI254 (ng/g dry)
f. T + 12,10 AUG 83
Figure 40. (Sheet 2 of 4)
92

-------
1.00-1

0.95-

0.90-

0.85-

0.80-
f
0.75-
'a
0.70-


1.00-
(/>
O
0.95-
JO

C
>s
0.90-
V)
c
4)
o
0.85-
u

o
0.80-
u

<1
0.75-

0.70-
1.00
0.95-
0.90-
0.85
0.80
0.75-
0.70-
o
q. T + 15, 6 SEP 83
~
~
ft
	1	1	r
h. T +21, 18 OCT 83
a
v
1-
0
O CNTR
400E
~ I000E
? REFS
4
-A
4000
1000	2000	3000
PCB AS AI254 (ng/g dry)
i. T + 27, 29 NOV 83
Figure 40. (Sheet 3 of 4)
93

-------
o
-0
E
c
V
Q.
O
u
U
<
I.OOn
0.95
0.90-
0.85-
0.80-
0.75-
0.70
I.OO-i
0.95-
0.90-
0.85-
0.80-
0.75-
0.70
A
	j	!	1	
j. T + 43, 20 MAR 84
	1	1	r—
k. T +55, 5 JUN 84
l.OO-

0.95-

0.90-


V
0.85-
o 2
at
0.80-

0.75-
ฆ
0.70-
1
f
i i
0
o CNTR
a 400 E
~ IOOOE
v REFS
	,	,	,	,	,	
1000	2000	3000
PCB AS AI254 (ng/g dry)
T + 116,13 AUG 85
Figure 40. (Sheet 4 of 4)
4
-4
4000
94

-------
Table 20
Adenine Nucleotide Concentrations (pmol/g Wet Weight) Measured in
N. inoisa Sampled on Specified Dates at the FVP
Field Stations
Station
400E
1000E
REFS
400E
1000E
REFS
400E
1000E
REFS
400E
1000E
REFS
400E
1000E
REFS
400E
1000E
REFS
ATP
0.61
	*
0.65
0.27
0.25
0.59
0.43
0.88
0.64
1.16
1.38
1.56
1.24
1.07
1.33
0.83
0.76
0.99
ADP
AMP
12 Apr 83 (T - 5)
0.16
0.05
0.24	0.06
06 Jun 83 (T + 3)
0.23
0.22
0.35
0.09
0.09
0.09
19 Jul 83 (T + 9)
0.35
0.25
0.36
0.46
0.41
0.26
0.45
0.45
0.51
0.49
0.38
0.51
0.13
0.09
0.08
08 Sep 83 (T + 16)
0.13
0.09
0.08
14 Dec 83 (T + 30)
0.20
0.20
0.18
15 Mar 84 (T + 43)
0.14
0.15
0.15
Total
0.82
0.95
0.59
0.56
1.03
0.91
1.22
1.08
1.74
1.87
1.88
1.89
1.72
2.02
1.46
1.29
1.65
AEC
0.36
0.55
0.63
0.62
0.72
0.58
0.61
0.72
0.80
0.85
0.90
0.73
0.73
0.76
0.69
0.68
0.70
(Continued)
* Not sampled.
95

-------
Table 20 (Concluded)
Station
ATP
ADP


AMP
Total
AEC


21 Jun
84
(T +
57)


400E
2.18
0.55


0.13
2.86
0.85
1000E
1.94
0.73


0.18
2.85
0.79
REFS
2.14
0.64


0.11
2.89
0.84


10 Oct
84
(T +
73)


400E
1.99
0.60


0.09
2.68
0.84
1000E
1.25
0.45


0.04
1.74
0.83
REFS
2.60
0.56


0.07
3.23
0.89


26 Jun
85
(T +
110)


400E
1.86
0.63


0.14
2.63
0.81
1000E
1.63
0.66


0.19
2.48
0.78
REFS
2.33
1.30


0.04
3.67
0.81

-------
adenine nucleotide pool concentrations and for AEC are presented graphically
in Figure 41. Because of seasonal influences on the organisms similar to
those described for M. edulis, the data are compared among FVP stations only
within sampling dates as was done for M. edulis. Limiting the comparisons of
field measurements to within a sampling period reduced sample size to a maxi-
mum of three when all stations sampled for N. incisa were sampled. Regression
analysis with this sample size is not appropriate; therefore, the data are
presented graphically to illustrate trends. Because detailed field exposure
data are not available, tissue concentrations of PCBs are used to represent
BRH exposure concentrations experienced by the organisms. It would be imprac-
tical to present graphs for all of the data for each of the nine sampling
dates. PCBs were selected because of the good relationship previously de-
scribed between tissue concentrations of PCBs and BRH exposure concentrations
in the laboratory experiment. The responses of total adenine nucleotide pool
concentrations and AEC in N. incisa are presented for comparison with the data
for M, edulis and because these responses represent two different ways of
treating adenine nucleotide data. The pool is the simple sum of all the
adenine nucleotide pools. The AEC is a weighted proportion of ATP in the
total pool. Neither of these effects measurements responded to BRH exposure
in laboratory experiments. When the field data were examined graphically,
there were trends among stations for adenine nucleotide pool concentrations
and for AEC on T + 16. There were no trends among stations on other dates.
Laboratory-to-Field Comparisons
Mytilus edulis
125.	The laboratory-to-field comparison was completed in two parts and
included both tissue residue and effects data. The approach taken was first
to establish whether exposure conditions were similar in the laboratory and
field residue data. Comparable tissue residues were interpreted as being in-
dicative of comparable BRH exposures. The second step was to compare the
adenine nucleotide pool concentrations and AEC values of the laboratory- and
field-exposed mussels with similar tissue residue concentrations.
126.	Residues. Results of the cluster analysis suggested several gen-
eral observations. First, the samples that were most similar included all the
field residues collected after T + 2 and the laboratory 0-percent BRH
97

-------
1.0
0.9
0.8-
0.7-
0.6-
0.5-
0.4
0.3H
- 0.2
0)
i 0.1
E
>>

c
0>
a.
_o
O
Ui
<
O 0% BRH
o
50/50
100%
BRH o
r2.5
2.4
2.3
2.2
2.1
L2.0
-1.9
-1.8
.7
200 400 600
a. Laboratory
800
i i	1	1	1	r
1000 1200 1400
1600
1.0 1
0.9-
0.8
0.7
0.6-
0.5-
0.4-
0.3-
0.2
0. H
• REFS
A 400E
ฆ I000E
r1-1
- 1.0
-0.9
-0.8
-0.7
-0.6

O
>>
M
T3
V
to
0
<_>
1
a>
J
o>
o
ฃ
CO
U)
a
i-
o
LU
-J
o
3
Z
u
z
z
UJ
a
<
200 400 600 800 1000 1200
PCB AS AI254 (ng/g)
b. T-5 , 12 APR 83
Figure 41. Relationship between total adenine nucleotide
concentration and AEC in N. incisa and PCB tissue residue
concentrations in laboratory- and field-exposed animals.
The laboratory data are presented to provide a perspec-
tive between the residue concentrations of laboratory-
and field-exposed worms (Sheet 1 of 4)
98

-------
o
I i i i i i I i i r
c. T + 3, 6JUN83	# REFS
A400E
I0-|	BIOOOE
0.9-
0.8
0.7
0.6-1 •	D
0.5-
0.4-
0.3-
0.2H
0.1
-1.0
-0.9
-0.8
•0.7
ฆ0.6
rl.3
ฆ1.2
-I.I
-1.0
0.9
0.8
—i	1	1	1	1	1	1	1	1	1	1
200 400 600 800 1000 1200
PCB AS AI254 (ng/g)
d. T+9, 19 JUL 83
Figure 41. (Sheet 2 of 4)
99

-------
• REFS
A 400E
ฆ IOOOE
i	i	r	i	r
e. T+16, 8 SEP 83
o
200 400 600 800
PCB AS AI254 (ng/g)
f. T+30, 30 DEC 83
r20

- 1.9

- 1.8




in
-1.5
o

u
1
T
"5

>.

ฆo

o>

N

o

E

3
ฆ2.1


LU

Q
ฆ2.0
1-

O

LJ
-1.9
_1
O

3

Z
-1.8
UJ

z
ฆ1.7
z
Hi

o

<
-1.6
-1
<

~—

o

t-
—i	1	1	
1000 1200
Figure 41. (Sheet 3 of 4)
100

-------
o
• REFS
a 400E
ฆ IOOOE
—i	1	1	1	1—
g. T + 43, 15 MAR 84
r 1.7
1.6
1.5
-1.4
-1.3
1.2
u>
o
-O
E
>.

-------
exposures. This would indicate that mussels in the field received minimal ex-
posure to BRH material after the initial disposal operation. Second, mussels
collected predisposal (CNTR, 400E, 1000E) and those collected shortly after
disposal (1000E at T + 0 and T + 2) were more similar to the other field sam-
ples than the laboratory samples. This would imply that, even during dispo-
sal, BRH exposures at these stations were more similar to subsequent postdis-
posal field residues than to laboratory BRH exposures. Third, all of the
laboratory residues were more similar to each other than any of the field sam-
ples. 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 the
other field exposures. 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 laboratory samples than the field.
127.	Effects. Analysis of the residue data suggested that the most
valid comparison between laboratory and field adenylate nucleotide 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 adenine nucleotide data
of mussels exposed to even 10-percent BRH in the laboratory to any mussels in
the field would not be appropriate because the residues were dissimilar. Ad-
ditionally, comparison of mussels collected from the field when environmental
conditions were not similar to those in the laboratory would not be
appropriate.
128.	Laboratory experiments were conducted in the spring at water tem-
peratures of 15ฐ C. Comparable field conditions existed at T + 0 and T + 2 in
the field. The AEC for mussels exposed to 0-percent BRH in the laboratory for
28 days was 0.83. The field-exposed mussels had AEC values of 0,87, 0.87, and
0.87 for the T + 0 (1000E, REFS) and T + 2 (1000E) collections, respectively.
These AEC values were indicative of metabolically active individuals in a non-
limiting environment. The only other collection that occurred in the spring
when water temperatures were similar was at T + 55. The AEC values for these
mussels were 0.89 and 0.88 for stations 400E and REFS, respectively. These
values are comparable with that value (0.83) for mussels exposed 28 days to
0-percent BRH in the laboratory. A comparison of the total adenylate nucleo-
tide pool data for these same samples also reveals no clear differences be-
tween laboratory and field data.
102

-------
129.	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 sediment exposures in the
field. In fact, the highest field residues were less than the residues of the
mussels exposed to the lowest BRH sediment concentration (1.5 mg/JL) in the
laboratory. Therefore, the data suggest that there was little or no overlap
in laboratory and field BRH sediment exposure concentrations. The resultant
effects data indicated adverse effects on mussels due to laboratory exposure,
whereas in the field there were no effects attributable to BRH sediment
exposure.
Nephtys inaisa
130.	The laboratory-to-field comparison was completed in two parts and
included both tissue residue and effects data. The approach taken was to es-
tablish first whether 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 adenine nucleotide pool concentrations and
AEC values of the laboratory and field worms at similar exposures.
131.	Residues. Data for the 12 representative chemical variables were
analyzed statistically by cluster analysis. The purpose of this procedure was
to identify distinctive patterns of association among the N. inaisa sampled
from laboratory experiments and field stations. The cluster analysis revealed
no consistent clustering of the laboratory data separate from field data.
This agrees with the overlapping range of residue data in laboratory and field
samples. The implication is that laboratory exposures to BRH material accu-
rately reflected the range of field exposures to BRH material for N. inaisa.
132.	Effects. The tissue residue data Indicated that comparison of
laboratory and field adenylate nucleotide data is valid. This comparison is
appropriate when environmental conditions in the field are similar to those in
the laboratory. Laboratory experiments were conducted with 20ฐ C seawater.
Comparable conditions existed at T + 8, T + 16, and T + 72 in the field. The
AEC for worms exposed to 0-percent BRH sediment in the laboratory for 42 days
was 0.85. The field-exposed worms had AEC values of 0.90, 0.85, and 0,80 for
T + 16 (REFS, 1000E, and 400E, respectively) and 0.89, 0.83, and 0.84 for
103

-------
T + 72 (REFS, 1000E, and 400E, respectively). The AEC values were very simi-
lar and were indicative of metabolically active individuals in a nonlimiting
environment. The AEC values for worms from the T + 8 collection could not be
used for comparison with other AEC values because of the extraction problems
experienced at this time. A comparison of total adenine pool data for these
same samples also revealed no clear differences between laboratory and field
worms.
Residue-Effects Comparisons
133. Regression analysis was used to determine whether any relationship
existed between the five biological measures (AEC, concentrations of ATP, ADP,
AMP, and total adenine nucleotide pools) and the tissue concentrations of the
selected 10 chemicals and 2 summary statistics for PAHs. The 5 biological
measures were regressed on the 12 chemical variables for the laboratory and
field data for both M. edulis and N. inoisa. This results in a total of 240
regression analyses. It is not reasonable to present all of this information
graphically in this report. Instead, these results are presented in
Tables 21-24. These tables contain statistical P values indicating degree
of statistical significance for each analysis. A P value of 0.05 or less
indicates a statistically significant relationship between the two variables
regressed together; a P value of 0.1 or less is useful for identifying pos-
sible trends in the data. The P values in Tables 21-24 were classified into
significance categories of 0.05 or 0.1, by biological variable, by species,
and by laboratory and field. This information is presented in Table 25. This
table facilitates comparison of the regression analysis results in three ways:
by biological variable, by species, and by laboratory versus field categories.
For example, the results in the grand total rows at the bottom of the table
permit ranking the biological variables from most to least correlated with
tissue concentrations of the 12 chemical variables as follows: adenine nuc-
leotide pool > total ATP > total ADP > AEC > total AMP. The results in the
grand total column facilitate comparison by species and by laboratory versus
field categories. For example, M. edulis had a total of 35 significant corre-
lations (P <. 0,05) out of a possible 120, while N. inoisa had a total of 14
significant correlations out of a possible 120. Clearly, the biological re-
sponses of M. edulis were more closely correlated with tissue concentrations
104

-------
Table 21
Summary of P Values Indicating Degree of Statistical Significance
for Each Regression Analysis Between Biological Variables and
Tissue Concentrations of Contaminants for Laboratory


Samples of
M. edulis





Biological
Variable





Total Adenine

Chemical Compound
ATP
ADP
AMP
Nucleotide Pool
AEC
Phenanthrene
0.282
0.424
0.608
0.063
0.405
Sum of 178 alkyl
0.007
0.590
0.323
0.006
0.298
homologs





Fluoranthene
0.007
0.429
0.314
0.005
0.314
Benzo(a)pyrene
0.030
0.571
0.670
0.001
0.688
SUM of PAHs
0.008
0.459
0.416
0.003
0.383
CENT of PAHs
0.616
0.689
0.485
0.655
0.688
PCB as A1254
0.008
0.669
0.272
0.019
0.235
Ethylan
0.010
0.670
0.269
0.023
0.234
Copper
0.012
0.477
0.472
0.006
0.393
Cadmium
0.152
0.232
0.635
0.001
0.681
Chromium
0.684
0.085
0.160
0.054
0.235
Iron
0.681
0.688
0.516
0.304
0.615
of the 12 chemical variables than was true for the biological responses of
N. inoi8a. A laboratory-versus-field comparison of the results revealed that
M. edulis had 15 significant correlations in the laboratory versus 20 in the
field, while N. inaiea had 13 in the laboratory versus 1 in the field. With
P ฆ 0.05 , out of 60 possible correlations, 3 can be expected by random chance
alone. For M. edulis, slightly more correlations occurred in the field than
in the laboratory. For N. inoisa, the correlations were limited almost en-
tirely to the laboratory.
134. These data may be examined also by chemical class. Table 26 pre-
sents the results of PAHs and metals by species and by laboratory versus field
105

-------
Table 22
Summary of P Values Indicating Degree of Statistical Significance
for Each Regression Analysis Between Biological Variables and
Tissue Concentrations for Field Samples of M. edutis
Biological Variable
Chemical Compound
ATP
ADP
AMP
Total Adenine
Nucleotide Pool
AEC
Phenanthrene
0.564
0.008
0.071
0.054
0.034
Sum of 178 alkyl
homologs
0.117
0.147
0.171
0.684
0.062
Fluoranthene
0.309
0.012
0.035
0.495
0.009
Benzo(a)pyrene
0.072
0.496
0.363
0.434
0.199
SUM of PAHs
0.087
0.229
0.201
0.641
0.085
CENT of PAHs
0.093
0.001
0.034
0.002
0.036
PCB as A1254
0.044
0.314
0.486
0.036
0.669
Ethylan
0.076
0.382
0.298
0.508
0.142
Copper
0.305
0.011
0.004
0.323
0.007
Cadmium
0.447
0.499
0.134
0.459
0.491
Chromium
0.162
0.007
0.004
0.460
0.005
Iron
0.401
0.014
0.018
0.344
0.018
categories. For M. edutis, the results indicate correlations between adenyl-
ate nucleotide concentrations and both PAH and metal concentrations in both
laboratory and field. For PAHs, there were eight significant correlations in
the laboratory and five in the field. For metals, there were three in the
laboratory and nine in the field. Overall, for M. edutis , the biological re-
sponse correlations with PAHs (13) were about the same as with metals (12).
For N. inoisa, the correlations between biological responses and contaminant
concentrations in tissues were limited almost entirely to PAHs in the labora-
tory (12 of 13).
106

-------
Table 23
Summary of P Values Indicating Degree of Statistical Significance
for Each Regression Analysis Between Biological Variables and
Tissue Concentrations of Contaminants for Laboratory
Samples of N. inoisa
Biological Variable
Chemical Compound
ATP
ADP
AMP
Total Adenine
Nucleotide Pool
AEC
Phenanthrene
0.006
0.042
0.626
0.002
0.050
Sum of 178 alkyl
homologs
0.059
0.056
0.490
0.032
0.196
Fluoranthene
0.102
0.066
0.480
0.059
0.281
Benzo(a)pyrene
0.001
0.044
0.691
0.0002
0.026
SUM of PAHs
0.024
0.049
0.514
0.012
0.119
CENT of PAHs
0.297
0.151
0.673
0.217
0.491
PCB as A1254
0.135
0.045
0.733
0.078
0.392
Ethylan
0.124
0.078
0.473
0.075
0.280
Copper
0.256
0.089
0.460
0.165
0.513
Cadmium
0.174
0.151
0.274
0.118
0.362
Chromium
0.978
0.314
0.920
0.822
0.519
Iron
0.965
0.214
0.703
0.788
0.482
107

-------
Table 24
Summary of P Values Indicating Degree of Statistical Significance
for Each Regression Analysis Between Biological Variables and
Tissue Concentrations for Field Samples of N. inei-sa
Biological Variable
Chemical Compound
ATP
ADP
AMP
Total Adenine
Nucleotide Pool
AEC
Phenanthrene
0.053
0.181
0.930
0.057
0.417
Sum of 178 alkyl
homologs
0.060
0.328
0.710
0.078
0.540
Fluoranthene
0.068
0.293
0.830
0.081
0.513
Benzo(a)pyrene
0.134
0.617
0.529
0.185
0.919
SUM of PAHs
0.078
0.378
0.651
0.102
0.639
CENT of PAHs
0.056
0.663
0.651
0.095
0.374
PCB as A1254
0.338
0.609
0.719
0.373
0.753
Ethylan*
—
—
—
—
—
Copper
0.720
0.955
0.200
0.812
0.501
Cadmium
0.074
0.115
0.076
0.091
0.292
Chromium
0.328
0.111
0.481
0.244
0.081
Iron
0.905
0.194
0.032
0.654
0.328
* Ethylan could not be quantified because of analytical interference.
108

-------
Table 25
Summary of the Number of Significant Correlations Between Adenine
Nucleotide Concentrations and Tissue Contaminant Concentrations
by Biological Variable, by Species, and by Laboratory
Versus Field Categories
Biological Variable

Lab/
Signif-



Total Adenine

Grand
Species
Field
icance
ATP
ADP
AMP
Nucleotide Pool
AEC
Total
M. edulis
Lab
0.05
7
0
0
8
0
15


0.10
7
1
0
10
0
18

Field
0.05
1
6
5
2
6
20


0.10
5
6
6
3
8
28

Total
0.05
8
6
5
10
6
35


0.10
12
7
6
13
8
46
N. inoisa
Lab
0.05
3
4
0
4
2
13


0.10
4
8
0
7
2
21

Field
0.05
0
0
1
0
0
1


0.10
6
0
2
5
1
14

Total
0.05
3
4
1
4
2
14


0.10
10
8
2
12
3
35
Grand
Total
0.05
11
10
6
14
8
49


0.10
22
15
8
25
11
81
109

-------
Table 26
Summary of the Number of Significant Correlations Between Adenine
Nucleotide Concentrations and Tissue Contaminant Concentrations
by Chemical Class, by Species, and by Laboratory
Versus Field Categories
Lab/
Species
Field
Significance
PAHs*
Metals*
M. edulis
Lab
0.05
8
3


0.10
9
5

Field
0.05
5
9


0.10
11
9

Total
0.05
13
12


0.10
20
14
N. inaisa
Lab
0.05
12
0


0.10
16
1

Field
0.05
0
1


0.10
7
5

Total
0.05
12
1


0.10
23
6
* Maximum possible number of significant correlations for PAHs ~ 25, for
metals - 20 for species, for laboratory or field category.
110

-------
PART IV: DISCUSSION
135.	The objectives of this study were to: (a) field verify laboratory
results and (b) investigate residue-effect relationships in M. edutis and
N. inoisa after exposure to BRH sediment in the laboratory and in the field.
The design of this study followed a logical progression from BRH exposure to
tissue residue concentration to biological effects. The discussion will par-
allel this approach by establishing the exposure-residue and residue-effect
relationships separately for the laboratory and for the field. This permits a
comparison of the laboratory and field results. Finally, the residue-effects
relationship will be considered in depth.
Laboratory Experiments
Mytilus edulis
136.	There was a strong link between exposure to BRH sediment and se-
lected tissue residues in M. edulis, as confirmed by the monitoring data col-
lected during the laboratory experiments. In addition, the relationship be-
tween 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/fc, 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.
137.	The strong exposure-residue relationships measured in the labora-
tory experiments indicate beyond reasonable doubt that the contaminants in BRH
sediments are biologically available. That mussels were affected by exposure
to BRH sediments is evident by reduced scope for growth, clearance (feeding)
rate, and shell growth rate in both experiments (Nelson et al. 1987).
Ill

-------
138.	In this report the responses of adenine nucleotides and AEC to
stress are considered. The central role of adenine nucleotides in energy
transformation and in metabolic regulation suggests their potential usefulness
as indicators of sublethal stress (Vetter and Hodson 1984). The adenine
nucleotides were measured in the adductor muscle tissue of M. edulis from all
treatments in laboratory Experiments 1 and 2. The exposed organisms from
these experiments were stressed as evidenced by reduced scope for growth in-
cluding reduced clearance (feeding) rates (Nelson et al. 1987). It might be
expected that under stress conditions and reduced food intake, the adenine
nucleotide pools of these organisms should be affected. Of the five variables
considered, ATP, ADP, AMP, adenine nucleotide pool, and AEC, only the pool
2
responded. This response was significant (P = 0.001, r = 0.94) only for the
day 14 data pooled from both experiments. The AEC measured in M. edulis did
not respond to BRH exposure in the laboratory.
139.	Caution should be exercised when interpreting adenine nucleotide
concentration data as indicators of stress. These concentration data repre-
sent "standing crop" or "static" pool sizes and convey little or no informa-
tion about energy flow. The turnover time for ATP in metabolically active
tissues may be less than 1 sec, and its concentration relative to energy stor-
age compounds is exceedingly low (Vetter and Hodson 1984). Therefore, ATP
does not represent an energy reserve but, rather, the ability of the cell to
generate energy. It is the rate of energy flow through ATP between anabolic
and catabolic processes that is of interest in measuring an organism's re-
sponse to stress. A stressor may affect metabolism by increasing the demand
for energy and/or by inhibiting the production of energy. If a stress causes
an increase in metabolism (and therefore energy flow) without exceeding the
organism's ability to regenerate ATP from energy reserves, there will be no
change in ATP concentration. Changes in ATP concentration will occur if a
stress causes a metabolic demand that exceeds the organism's ability to regen-
erate ATP; if the energy reserves are exhausted; or if the stress blocks the
ability to resynthesize ATP. AEC is a less sensitive indicator of stress be-
cause it may remain constant even when decreases in ATP result in decreases in
total adenylate concentrations rather than increases in ADP and AMP. Vetter
and Hodson (1984) state, "A change in total adenylate concentration is pre-
cisely what does occur to a greater or lesser extent in almost all organisms
studied to date." The response of M. edulis In this study to BRH exposure in
112

-------
the laboratory is consistent with this explanation. The total adenylate con-
centration in M. edulis muscle tissue decreased with exposure to increasing
concentrations of BRH sediment, while the AEC remained unchanged. Atkinson
(1977) proposed AEC as a biochemical concept and explained in great detail its
conservative and cybernetic nature. These very qualities make AEC insensitive
as an indicator of stress.
Nephtys inaisa
140.	The tissue residue concentrations of all the organic compounds
measured in N. inaisa increased with increasing exposure to BRH sediments.
This strong exposure-residue relationship measured in the laboratory experi-
ments indicates beyond reasonable doubt that the worms were exposed to BRH
sediments and that the contaminants in BRH sediments were biologically avail-
able. That worms were affected by exposure to BRH sediments is evident by re-
ductions in growth, increases in respiration rate, and exposure-related reduc-
tions in net growth efficiency (Johns and Gutjahr-Gobell 1988).
141.	Adenine nucleotides were measured in whole worms from all treat-
ments at all sampling times in three laboratory experiments. Exposure condi-
tions ranged from time zero, nonexposed, to 200 mg BRH/Jl for 42 days. Of the
five variables considered, ATP, ALP, AMP, adenine nucleotide pool, and AEC,
none exhibited a response to BRH exposure. Caution should be exercised when
interpreting apparent "no-effect" responses in adenine nucleotide concentra-
tion data. These data represent static or snap-shot measurements of adenine
nucleotide pool sizes and therefore convey little or no information about en-
ergy flow. Ideally, these measurements should be supported by measurements of
energy storage compounds such as lipid and glycogen. Vetter and Hodson (1984)
state:
Adenylate measurements alone fail to pinpoint a specific
cause and effect relationship between a type of pollutant
and an observed effect. This is not a failure of
adenylate measurements in particular but an unavoidable
consequence of the generalized stress response as it has
evolved in most organisms. Combining adenylate measure-
ments with other measures of energy reserves, such as
lipid and glycogen, can improve detection of low-level
chronic stress and begin to indicate the mechanism of
toxic action.
113

-------
Field Experiments
Mytilus eduli-s
142.	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.
143.	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 in the field). The following generalizations are evident from the esti-
mated BRH exposure concentrations: (a) the two independent methods, tissue
residue and whole water analysis, provided remarkably similar estimates of BRH
exposure; and (b) there was a distinct exposure signal at 1 m above the bottom
during and immediately postdisposal, and that signal was transient, decreasing
spatially and temporally postdisposal.
144.	Comparison of the tissue residue and water chemistry estimates of
BRH concentrations in CLIS indicated very good correspondence as demonstrated
by the following examples. Tissue residue data (PCBs) from the T + 2 col-
lection indicated that the BRH concentration was estimated to range between
1.4 and 0.8 mg/i at the CNTR station. Water samples from the same station es-
timated the BRH concentration to range between 1.1 and 0.7 mg/Jl using PCB val-
ues, and 1.3 and 0.7 using copper values. Approximately 1 month later, BRH
concentration estimates, based on PCB tissue residues, were between 0.7 and
0.2 mg/i. at the CNTR station. The corresponding water chemistry estimates of
BRH concentrations at the CNTR station ranged from 0.2 to 0.1 mg/H, using
PCBs, and 0.6 to 0.3 mg/ฃ, using copper. The similarity of these estimates
indicates that exposure based on whole water chemistry concentrations and tis-
sue residue concentrations tracked reasonably well.
145.	The good relationship between residue and water chemistry esti-
mates of BRH concentrations further supports the second generalization men-
tioned above, that, BRH exposure 1 m above the bottom was maximal immediately
postdisposal and decreased over time. Spatially, both the PCB tissue residue
114

-------
and water chemistry data indicated that BRH exposure decreased moving away
from the CNTR station immediately postdisposal. 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 dis-
posal 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/Ji (tissue residues at T + 2, Table 15). At the time of the
next collection, this value decreased by approximately one half and continued
to decrease over time.
146.	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 concen-
tration at the REFS station, was assumed to remove the background concentra-
tion 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 decreased rap-
idly following completion of the disposal operation.
147.	Two weeks postdisposal (T + 2), when maximum tissue residues
(i.e., exposures) occurred, no decreases in AEC or any of the adenine nucleo-
tide concentrations were observed. One explanation is that the BRH exposure
concentration at T + 2 may have been insufficient to cause a response. The
estimated maximum BRH exposure at a station where mussels were deployed
(0.8 mg/ฃ was about half that of the lowest suspended sediment concentration
tested in the laboratory (1.5 mg/H). The response threshold for the adenine
nucleotide was >5 mg/JI BRH suspended sediment. Therefore, the BRH signal in
the field was "weaker" than that required to elicit a response in the labora-
tory experiments. Based upon laboratory exposure-response data, one would
have predicted that for the measured field exposures no effects would occur to
the adenine nucleotides. This is in fact exactly what was observed.
148.	AEC and adenine nucleotide concentrations for mussels deployed in
CLIS for 1 month and longer were affected only by natural phenomena. Mussels
115

-------
collected during August and September, regardless of location, had AEC values
Indicative of stress. This phenomenon has been reported for mussels previ-
ously by Skjoldal and Barkati (1982) and Zaroogian et al. (1982) and is prob-
ably due to a combination of reproductive and temperature (22ฐ C) stresses.
149.	The final collection (T + 116), a 1-month deployment, indicated
that AEC was not different among stations; however, the AEC values were higher
than the AEC for these same mussels at the time of deployment. This was due
to an abnormal occurrence that was endemic to Narragansett Bay, including the
site of the reference population, at the time mussels were collected for de-
ployment in CLIS. A bloom of a small (<2.0 y) algal species occurred through-
out Narragansett Bay. This alga, present in concentrations greater than
1.0 billion cells/J,, caused the mussels in the Bay to cease feeding, resulting
in mass mortalities in mussel populations. As a result, mussels deployed for
T + 116 were not in the best condition, and deployment in CLIS appeared to be
therapeutic.
Nephtys inaisa
150.	Tissue concentrations of PCBs in N. incisa increased at all FVP
stations during the summer of 1983 and reached their highest measured concen-
trations in September. This evidence indicates that exposure to dredged mate-
rial at the sediment-water interface continued throughout the CLIS study area
during the summer of 1983. Since there were no significant storms during the
summer of 1983, the contaminant exposures were probably due to initial disper-
sion of dredged material and tidally driven resuspension and movement of sedi-
ments from the dredged material mounds.
151.	There were difficulties with adenylate extraction from N, incisa
during the first several months of field sampling. These problems were re-
solved by September 1983 (Zaroogian et al. 1985). Therefore, the data from
September 1983 (T + 16) could be tested for spatial differences. Differences
among stations were found to be significant for all adenine nucleotide concen-
trations and AEC. The differences in AEC were highly significant (P = 0.0001)
with the lowest values closest to the disposal mound: 400E (0.80), 1000E
(0.85), and REFS (0.90). Although these differences were statistically sig-
nificant, the AEC values were indicative of nonstressed organisms.
152.	During the winter months (T + 29, T + 43), what appeared to be a
seasonal stress occurred. This stress appeared to be greater than any stress
induced by disposal operations. Davis (1979) reported a sixfold to sevenfold
116

-------
reduction in respiration in N. incisa during winter months (seawater tempera-
ture 0ฐ C) when compared with respiration during the summer months (seawater
temperature 24ฐ C). Seawater temperatures recorded at the December 1983 and
March 1984 samplings were 7ฐ C and 0.9ฐ C, respectively. Thus, the AEC values
for N. inaisa during winter months, which were indicative of metabolic stress,
could have been due to lowered respiration rates caused by low seawater
temperatures.
Laboratory-to-Field Comparisons
153.	A primary objective of the FVP was to field verify the laboratory
biological responses by measuring the same response under both laboratory and
field exposures. A basic and often implicit assumption is that results de-
rived from laboratory tests are directly applicable in the field. This study
was designed to test that assumption.
Mytilus edulis
154.	Exposure conditions must be examined to determine whether the bio-
logical responses are responding to comparable exposures in the laboratory and
in the field. A comparison of laboratory and field data indicated one obvious
fact: laboratory and field BRH exposures were different. The two independent
estimates of BRH exposure indicated that maximum exposures in the field were
half that of the lowest BRH exposure in the laboratory. Cluster analysis of
laboratory and field mussel residue data yielded results similar to those ob-
tained in estimating the field exposures. That is, the mussel residues in the
field were most similar to mussels exposed to reference sediment,in the labo-
ratory. Considering these data, discussion of the laboratory-field comparison
will focus initially on when conditions (exposures and residues) were similar
in the laboratory and field, and secondly on estimated concentrations of BRH
suspended material required to affect concentrations of adenine nucleotides
and AEC values.
155.	The exposure and residue data indicated that the most legitimate
comparison between laboratory and field biological effects 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 biological effects data should provide the most accurate
117

-------
laboratory-to-field comparison. The AEC value for mussels exposed to
0-percent BRH sediment in the laboratory for 28 days was 0.83. The field-
exposed mussels exhibited AEC values of 0.87, 0.87, and 0.87 for the T + 0
(1000E), T + 0 (REFS), and T + 2 (1000E) collections, respectively. The simi-
larity of these AEC values indicated that the relative physiological condition
of these mussels was the same when environmental conditions were most alike.
The mussel collection at T + 55 also occurred in the spring at water tempera-
tures similar to the laboratory exposures; however, these mussels had been
deployed in CLIS for 8 months. The AEC values of these mussels, 0.87, 0.83,
and 0.88 for 400E, 1000E, and REFS, respectively, were similar to the AEC for
mussels in the 1-month laboratory exposure (0.83).
156.	The purpose of this qualitative comparison between laboratory and
field was to determine 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
concentrations, food quantity, etc.), the AEC values and the adenine nucleo-
tide concentrations between the two exposures were remarkably similar.
157.	A second aspect of the laboratory-to-field comparison was a qual-
itative evaluation of the estimate of BRH material required to produce an ef-
fect on AEC and the adenine nucleotide concentrations in the laboratory and
field. The total adenine pool concentration for mussels in the laboratory
experiments provided a clear signal that exposure to 5 to 10 mg/ฃ of BRH mate-
rial for 14 days negatively affected M. edulis. From these data an estimated
BRH exposure ^5 mg/ฃ should be sufficient to adversely affect mussels exposed
in the field. The estimated maximum exposure concentration in the field
(0.8 mg/H), approximately half that in the lowest laboratory BRH exposure
(1.5 mg/ฃ), did not appear to affect the mussels. The mussels were exposed to
much higher concentrations of BRH (10x) in the laboratory than in the field.
The response in total adenine pool concentrations was elicited only at these
higher concentrations and therefore would not be expected in the field.
Nephtys inoisa
158.	Exposure conditions must be examined to determine whether the bio-
logical responses are responding to comparable situations in the laboratory
and in the field. Physical data were used to make three different estimates
of exposure to BRH material at the FVP stations. Water chemistry data were
used to estimate milligrams of BRH per litre 1 m above the bottom at the FVP
118

-------
stations (Nelson et al. 1987). With the assumption of a 10x enrichment from
the 1 m above the bottom value, there is a predicted exposure at the sediment-
water interface of 6 to 13 mg BRH/ฃ at the FVP stations as a result of dis-
posal at the FVP site. Estimates of exposure via resuspension of surficial
sediments predicted much higher concentrations. A worst case estimate assumes
that all of the predicted suspended solids are BRH material from the disposal
mound. This estimate predicts up to 100 mg BRH/ฃ under quiescent conditions
and up to 300 mg BRH/ฃ under storm conditions. A more probable estimate as-
sumes that sediments resuspended at each station are the source of contami-
nants for the suspended solids. The estimate predicts a graded exposure at
the FVP stations with maximum values of 40 mg/Jl at 200E, 12 mg/ฃ at 400E, and
4 mg/I at 100E for quiescent conditions. Those values increase to 120 mg/ฃ at
200E, 40 mg/ฃ at 400E, and 10 mg/ฃ at 100E for storm conditions.
159.	If it is assumed that tissue concentrations in N. inaiaa are di-
rectly related to exposure concentrations, this relationship may be used to
test the reasonableness of the exposure predictions. This assumption is rea-
sonable, based on results from laboratory experiments. A cluster analysis of
all N. incisa tissue residue data revealed no consistent clustering of the
laboratory data separate from the field data. Therefore, if it is assumed
that tissue concentrations reflect exposure concentrations, then this associa-
tion of laboratory and field tissue concentration data indicates an overlap of
laboratory exposure conditions with field exposure conditions. The estimates
of field exposures to BRH sediment (milligrams per litre) suspended at the
sediment-water interface based on PCB concentrations in field-collected
N. incisa are up to 12 mg/ฃ at REFS, 88 mg/ฃ at 100E, and 30 mg/ฃ at 400E.
160.	Assuming that exposures were due to initial dispersion of BRH sed-
iments during disposal and subsequent resuspension and movement of sediments
from the dredged material mound, a combination of estimates seems appropriate.
The estimate based on water chemistry predicts exposures of at least 6 mg/ฃ at
the sediment-water interface at all FVP stations during disposal activities in
CLIS. The worst case resuspension estimate predicts exposures of up to
100 mg/ฃ in the vicinity of the disposal mound. These estimates (6-100 mg/ฃ)
agree well with those predicted by the tissue concentration exposure concen-
tration relationship (12-130 mg/I). The two laboratory exposures for the sis-
ter chromatid exchange response were 0 and 200 mg BRH/i, as suspended solids.
These exposures overlap the estimated range of exposures in the field,
119

-------
simulated clean control conditions at REFS and worst case storm conditions
near the disposal mound.
161.	In a prior laboratory study (Zaroogian et al. 1985), N. inaisa
were allowed to burrow into either 100-percent REF or 100-percent BRH sediment
prior to treatment with suspended REF or BRH sediments at concentrations up to
200 mg/ฃ. These experiments lasted for 10 days, and no differences in AEC oc-
curred among treatments. Thus, the results of this prior study were in agree-
ment with the present study where no differences in AEC or adenine nucleotide
concentrations occurred among treatments. The longest experiment in the pres-
ent study was 6 weeks.
162.	In contrast to the laboratory results where no differences were
reported among treatments, field studies showed highly significant differences
among stations in September 1983 (T + 16 weeks) for AEC and all adenine nucle-
otide concentrations. This response in the field coincides with peak expo-
sures as evidenced by highest tissue concentrations of PCBs measured in field
N. inaisa. There is marked difference between laboratory and field responses
in spite of estimated comparable exposures to BRH sediments. For example, the
highest concentration of PCB measured in tissues of laboratory N. inaisa was
1,500 ng/g dry weight (sampled at 42 days, 100-percent BRH treatment), while
the highest concentration of PCB measured in tissues of field N. inaisa was
1,250 ng/g dry weight (sampled 6 September 1983 at 400E) . A major difference
between laboratory and field studies was the duration of exposures: 6 weeks
in the laboratory versus 16 weeks in the field. The importance of exposure
time suggests that the PAHs may be affecting the adenine nucleotide concentra-
tion responses. The residue-effect relationships between tissue concentra-
tions of PAHs and adenine nucleotide concentrations responses in laboratory
N. inaisa further support this suggestion. These compounds may be metabolized
to reactive intermediates. A key enzyme system in this metabolism is the
cytochrome P-450 dependent mixed-function oxygenase (MFO) system. These en-
zymes are not functioning at all times. Their reactivity is induced, that is,
greatly increased, by exposure to substrate compounds (PAHs). This induction
is not immediate. Fries and Lee (1984) report an induction time of 4 to
8 weeks for the MFO system in the marine polychaete Nereis virens exposed to
benzo(a)pyrene.
163.	The contrasting patterns of increased tissue concentrations of
PCBs and PAHs in N. inaisa during the summer of 1983 indicate that the MFO
120

-------
system in the exposed worms had been activated. PCB tissue concentrations
reached a peak in September, 4 months postdisposal, indicating a continuous
exposure to contaminated sediments during this period. In contrast to the
4-month period of PCB increase, the highest PAH tissue concentrations occurred
in July, only 2 months postdisposal. Laboratory bioaccumulation data suggest
that N. inoisa can metabolize PAHS and that this metabolic capability was
induced during the 42-day experimental period. These data suggest that metab-
olism of PAHs was induced and was causing a sharp decline in tissue PAH con-
centrations despite continuous exposure to these compounds. Presumably, by
16 weeks postdisposal, the worm MFO system had been induced, resulting in a
significant biological response.
164.	The results of this study suggest that the activity of the MFO
system in N. inoisa may be an important variable because of the high concen-
trations of compounds, such as PAHs in BRH sediment, metabolized by these
enzymes. The MFO system is induced by exposure to contaminants such as PAHs.
Because this induction takes weeks in polychaetes, the laboratory experiments
were probably not long enough to induce maximum MFO activity. The shortness
of laboratory experiments with N. inoisa in the FVP program was cited as a
limitation also in the cytogenetic studies (Pesch et al. 1987). The simplest
explanation of why the laboratory and field results do not match for N. inoisa
in this study is that the laboratory experiments were not long enough. The
field data suggest that the laboratory experiments should be conducted for pe-
riods of at least 4 months. The results suggest also that MFO activity should
be monitored in organisms exposed to material such as BRH sediments.
Residue-Effects Comparisons
165.	Bioaccumulation of contaminants depends on various pharmacokinetic
processes including uptake, distribution, metabolism, and elimination. The
extent to which a contaminant will bioaccumulate depends on the relative rates
of uptake versus rates of metabolism and elimination. A contaminant that is
metabolized and eliminated rapidly, for example, probably will not
bioaccumulate.
166.	Bioaccumulation data provide evidence that contaminants are bio-
logically available. This is useful information for complex wastes such as
BRH sediments. However, bioaccumulation data alone provide no evidence of an
121

-------
effect or consequence to the organism. The relevance of a specific tissue
residue to the fitness of an organism is unknown.
167.	In this study, an attempt was made to identify relationships be-
tween tissue concentrations of selected BRH sediment contaminants and biologi-
cal effects under both laboratory and field exposures. It was assumed that
these selected contaminants were biologically available and had toxicological
properties that would affect adenine nucleotide concentrations in N. inaisa
and M. edulis. The difficulty was that BRH sediment contained many contami-
nants, any of which alone could be toxic to these test species and possibly
more toxic in combination. Thus, it is impossible to determine if the accumu-
lation of any of the selected contaminants was responsible for any observed
effect. Therefore, any relationship between body burden and biological re-
sponse can be attributed only to the sediment, although inferences or trends
may be identified with individual contaminants.
168.	Adenine nucleotides and AEC are of interest in measuring stress
effects because of their central role in energy transformation and their
importance as regulators of metabolic processes. Vetter and Hodson (1984)
state that the metabolic costs of stress may result from either:
(1) diverting assimilated energy to nonproductive func-
tions such as increased respiration, cell repair, cleaning
and avoidance activity or (2) decreasing the efficiency of
energy transfer reactions through toxic effects on enzyme
systems, changes in membrane potentials or genetic damage.
Either mechanism results in an increase in the dissipation
of assimilated energy as heat and a concomitant decrease
in growth and reproductive potential.
This study was not designed to distinguish between these mechanisms. There-
fore, the adenine nucleotide and AEC responses measured in M. edulis and
N. inaisa could be due to either or both mechanisms.
Mytilus edulis
169.	The strong exposure-residue relationships measured in the labora-
tory experiments indicate that the contaminants in BRH sediments are biologi-
cally available. Regression analysis was used to determine whether any rela-
tionships existed among the five biological measures (AEC, concentrations of
ATP, ADP, AMP, and total adenylate nucleotide pools) and tissue concentrations
of the selected 10 chemicals and 2 summary statistics for PAHs.
170.	In the laboratory, significant correlations existed between total
122

-------
adenylate pool and sum of 178 alkyl homologs, fluoranthene, benzo(a)pyrene,
sum of PAHs, PCB as 1254, ethylan, copper, and cadmium. This represents a
broad spectrum of chemicals, including several classes of organic compounds
plus metals. It is interesting to note that, in these same experiments, sig-
nificant correlations existed between scope for growth responses and the exact
same list of chemicals, (Nelson et al. 1987). Scope for growth, as with the
adenine nucleotide pools, is a measure of energy utilization. Thus, these two
independent measures suggest that these chemicals affected energy flow and
utilization in exposed M. edulis.
171.	Detoxification systems for a wide range of chemical compounds are
known to exist in marine organisms. An important feature of these systems is
that they are inducible; that is, their functional activity is greatly in-
creased by exposure to substrate chemicals. Therefore, organisms having these
systems are able to acquire tolerance to and the ability to excrete a variety
of chemicals. The primary detoxification systems so far identified are the
MFO system for organic xenobiotics and the metallothioneins for metals. These
systems apparently are universally distributed, and M. edulis has both MFO
(Livingstone 1985) and metallothionein (Roesijadi 1982, 1986). These systems
function to sequester contaminants and protect against direct toxic action.
Theoretically, an organism is protected until the detoxification systems
exceed saturation. Only when contaminants "spill over" do they have direct
toxic impact on an organism. However, as Jenkins and Brown (1984) point out,
this approach "ignores the potential indirect cost of detoxification to an or-
ganism. This cost may represent the energy required for the synthesis of de-
toxification proteins, competition for amino acids or the physical cost of ac-
cumulating large quantities of proteins and/or membrane-bound vesicles within
a cell."
172.	Declines in adenine nucleotide energy pools and scope for growth
indices (Nelson et al. 1987) indicate that M. edulis was stressed by exposure
to BRH sediments in the laboratory experiments. This stress was characterized
by increased energy costs. Measurements of the adenine nucleotide concentra-
tions can help to characterize the energy costs incurred by organisms under
stressful conditions. "However, the information content of the basic measure-
ments can be vastly Increased and the mechanism of toxic action can be sug-
gested if the changes in adenylate concentration are considered in the light
of changes in other energy compounds such as glycogen and neutral lipid
123

-------
reserves," state Vetter and Hodson (L984). Ultimately, energy costs of stress
need to be related to changes In growth and reproduction and to impacts on
populations.
173.	In contrast to the laboratory results, there were no strong
exposure-residue relationships measured in the field. Cluster analysis of
laboratory and field mussel residue data yielded results similar to those ob-
tained in estimating field exposures. Maximum exposures in the field were
half that of the lowest BRH exposure in the laboratory. These exposures
existed during disposal of BRH sediments at the FVP site and for several
months postdisposal. Thereafter, there was no measurable accumulation of con-
taminants in M. edulis tissues, indicating little or no exposure to BRH
sediments in the field during the 2 years of postdisposal monitoring. The
contaminant residues in the field mussels were most similar to those of mus-
sels exposed to reference sediment in the laboratory. Thus, the exposure data
and the tissue residue data indicate that in the field M. edulis was exposed
to low and transient levels of BRH sediments. The effects data support this
hypothesis.
174.	Use of adenine nucleotides as indicators of stress presumes that
the three adenylates remain almost constant within an organism except under
conditions of extreme energy consumption or of metabolic inhibition. When the
concentrations of the three adenylates change in response to stress, these
changes occur in consistent and predictable ways. This concept suggested that
the ratios of adenylate concentrations, as represented by AEC, were a useful
measure of an organism's well-being. As a response to stress, it is expected
that ATP concentration will decrease, AMP and ADP concentrations will in-
crease, and, therefore, AEC will decrease. The residue-effect data for
M. edulis from the field are inconsistent with these expectations. If in-
creased tissue concentrations of contaminants are to be construed as stress-
ful, then concentrations of ATP should be negatively correlated, concentra-
tions of ADP and AMP should be positively correlated, and AEC should be
negatively correlated. The field data for M. edulis show an opposite pattern.
Concentration of ATP was positively correlated with tissue concentrations of
PCB. Concentrations of ADP and AMP were negatively correlated with tissue
concentrations of five contaminants. AEC values were positively correlated
with tissue concentrations of five contaminants. The apparent AEC response to
iron further confirms the inconsistent nature of these data. Iron is present
124

-------
in all sediments in relatively high concentrations. It is not specific to BRH
sediments. Also, it is relatively nontoxic. It was included in the list of
12 chemical variables as a negative control. That is, detection of any bio-
logical response to iron concentrations was not expected. The adenine nucleo-
tide concentrations and AEC values indicate that FVP field-collected M. edulis
may have been responding to something, but that something was not tissue con-
centrations of contaminants, nor was it exposure to BRH sediments.
Nephtys inoisa
175.	The strong exposure-residue relationships measured in the labora-
tory experiments indicate that the contaminants in BRH sediments are biologi-
cally available. Regression analysis was used to determine whether any rela-
tionships existed between the five biological measures (AEC, concentrations of
ATP, ADP, AMP, and adenine nucleotide pools) and the tissue concentrations of
the selected 10 chemicals and 2 summary statistics for PAHs.
176.	Significant correlations were detected between the following re-
sponses and chemical contaminants: the adenine nucleotides correlated with
PAHs, phenanthrene, sum of 178 homologs, benzo(a)pyrene, and the sum of the
PAHs; measures of ATP correlated with PAHs, phenanthrene, benzo(a)pyrene, and
the sum of the PAHs; ADP showed similar correlations as ATP but with the addi-
tion of PCBs; finally, AEC correlated with phenanthrene and benzo(a)pyrene.
This represents a narrow spectrum of chemicals; 12 of 13 significant correla-
tions were with PAHs. In contrast, the physiological measure of net growth
efficiency responded to a broad spectrum of chemicals, including several
classes of organic compounds plus metals in these same laboratory experiments
(Johns and Gutjahr-Gobell 1988). The data suggest that the adenine nucleotide
pools in N. inoisa were affected by the PAHs in BRH sediment.
177.	Ironically, a direct correlation between tissue concentrations of
PAHs and a biological response is not always expected. For high molecular
weight PAHs, it is not the parent PAH compounds that are biologically damag-
ing. Damaging action is associated with metabolic products. These metabo-
lites inflict damage when the affected organisms are actively depurating.
Therefore, a simple correlation between tissue concentration of parent PAH
compounds and biological response will not exist generally.
178.	PAHs are metabolized by the MFO system. Polychaetes are known to
have active, inducible MFO systems (Lee et al. 1981; Lee and Singer 1980).
Induction of these enzymes takes weeks to months (Fries and Lee 1984). It is
125

-------
during maximum activity of the MFO system that maximum biological effect would
be expected. It is instructive on this point to compare laboratory and field
residue-effects data for the adenine nucleotide responses. This comparison is
complicated somewhat by technical difficulties encountered during the early
field measurements (Zaroogian et al. 1985). However, with this constraint,
some general observations may be useful. In the field there were no meaning-
ful significant correlations. The single exception, iron, was included as a
negative control. There were no correlations between tissue concentrations of
PAHs and any measure of adenine nucleotide response. Yet, at 16 weeks post-
disposal, there were highly significant differences in adenine nucleotide re-
sponses among field stations. In the laboratory, there were significant cor-
relations between tissue concentrations of PAHs and adenine nucleotide
responses. Yet, in the laboratory, there were no significant differences in
adenine nucleotide responses among treatments. Exposure data indicate compa-
rable exposures in the laboratory and in the field. The big difference be-
tween laboratory and field was length of exposure. The maximum length of lab-
oratory exposure was 6 weeks. The maximum response in the field was at
16 weeks. The data suggest that, in the laboratory at 6 weeks, depuration had
been initiated, residue-effect correlations had been established, but no sig-
nificant biological responses had been evoked. In the field, at 16 weeks,
depuration had actively depleted most of the tissue concentrations of PAHs,
residue-effects correlations were no longer present, and significantly differ-
ent biological responses among stations were present as a consequence. If
this interpretation of the data Is accurate, it suggests that the laboratory
experiments were not long enough. More time was needed to induce fully the
MFO system in N. incisa. The induction of the MFO system is crucial to an
organism's response to complex materials, such as BRH sediments, that contain
high concentrations of PAHs.
179. Overall, it appears that a number of factors other than BRH sedi-
ments were affecting the AEC response in the field. Even where statistically
significant responses were observed, they were indicative of nonstressed or-
ganisms and probably have little, if any, ecological significance.
126

-------
PART V: CONCLUSIONS
180.	There were three primary objectives in the FVP. The first objec-
tive was to test the applicability of biological responses to measure effects
of dredged material. The second objective was to field verify responses ob-
served in the laboratory and determine the accuracy of the laboratory predic-
tions. The third objective was to determine the degree of correlation between
tissue residues accumulated from dredged material and biological responses as
observed in both the laboratory and the field.
181.	The biological responses evaluated in this report included the
adenine nucleotide measures of ATP, ADP, AMP, adenine nucleotide pool, and
AEC. These responses were measured in M. edulis and N. inoisa exposed to BRH
sediment in the laboratory and in the field. The only significant laboratory
response was a reduction in total adenylate pool concentration measured in
M. edulis at BRH exposure concentrations higher than any estimated exposures
in the field. The only significant field responses were station-related
changes in all adenylate nucleotide concentrations measured in N. inoisa
16 weeks postdisposal. This represented an exposure lasting 10 weeks longer
than the longest laboratory exposure, which was 6 weeks. The field exposures
were indicative of nonstressed organisms and appeared not to result from ex-
posure to BRH sediments.
182.	The adenine nucleotide pool concentrations in organisms exposed to
BRH sediments will respond in a concentration-related manner. However, these
responses are relatively insensitive in M. edulis and are related to long ex-
posure periods in N. incisa.
183.	The apparent differences between laboratory and field responses
for M. edulis and for N. inoisa could be explained by differences in exposures
between the two situations. For M. edulis, the exposures were much higher in
the laboratory. For N. inoisa, the exposures in the laboratory and the field
were comparable, but the field response was significant at 16 weeks post-
disposal. This length of exposure far exceeded the length of any laboratory
experiments.
184.	Adenine nucleotide and AEC are important in energy transformation
and in regulation of metabolic processes. Therefore, it is not surprising
that responses in adenine nucleotide pools correlate with tissue concentra-
tions of BRH contaminants in exposed organisms. Measurements of the adenine
127

-------
nucleotide concentrations may help to characterize the energy costs incurred
by organisms under stressful conditions.
128

-------
REFERENCES
Adam, H. 1963. "Adenosine-5'-Phosphate: Determination with Phosphoglycerat
Kinase," Methods of Enzymatic Analysis, H. V. Bergmeyer, ed., Verlag Chemie
Weinheim, Academic Press, New York.
American Society for Testing and Materials. 1980. "Standard Practice for
Conducting Acute Toxicity Tests with Fishes, Macroinvertebrates, and Amphib-
ians," ASTM E-729-80, Philadelphia, Pa.
Atkinson, D. E. 1971. "Regulation of Enzyme Function," Annual Review of
Microbiology, Vol 23, pp 47-68.
	. 1977. Cellular Energy Metabolism and Its Regulation, Academic
Press, New York.
Atkinson, D. E., and Walton, G. M. 1967. "Adenosine Triphosphate Conserva-
tion in Metabolic Regulation; Rat Liver Citrate Cleavage Enzyme," Journal of
Biological Chemistry, Vol 242, pp 3239-3241.
Ball, W. J., and Atkinson, D. E. 1975. "Adenylate Energy Charge in
Sacahavomyces cerevisiae During Starvation," Journal of Bacteriology, Vol 121
pp 975-982.
Behm, C. A., and Bryant, C. 1975. "Studies on Regulatory Metabolism in
Moniezia expansa: The Role of Phosphofructo kinase (with a Note on Pyruvate
kinase)," International Journal of Parasitology, Vol 5, pp 339-346.
Bergmeyer, H. V. 1965. "Cell and Tissue Disintegration," Methods of
Enzymatic Analysis, H. V. Bergmeyer, ed., Academic Press, New York.
Bohlen, W. F., and Winnick, K. B. 1986. "Observations of Near-Bottom Sus-
pended Material Concentrations at the FVP Site, Central Long Island Sound
Dredged Materials Area: Immediate Predisposal, Disposal and Immediate Post-
disposal Period, April 18, 1983 - June 29, 1983," Science Applications Inter-
national Corporation, Newport, R. I., Marine Sciences Institute, Groton, Conn
Boyle, E. A., and Edmond, J. M. 1975. "Determination of Trace Metals in
Aqueous Solution by APDC Chelate Co-precipitation," Analytical Methods in
Oceanography, American Chemical Society, pp 44-55.
Chapman, A. G., Fall, L., and Atkinson, D. E. 1971. "Adenylate Energy Charg
in Escherichia coli During Growth and Starvation," Journal of Bacteriology,
Vol 108, pp 1072-1086.
Davis, W. R. 1979. "The Burrowing, Feeding, and Respiratory Activities of
Nephtys incisa Malmgren, 1865 (Polychaeta:Annelida)," Ph.D. Dissertation,
University of South Carolina.
Dean, D., and Mazurkiewicz, M. 1975. "Methods of Culturing Polychaetes,"
Culture of Marine Invertebrate Animals, W. L. Smith, and M. H. Chanley, eds.,
Plenum Press, New York.
Fries, C. R., and Lee, R. F. 1984. "Pollutant Effects on the Mixed Function
Oxygenase (MFO) and Reproductive Systems of the Marine Polychaete Nereis
virens," Marine Biology, Vol 79, pp 187-193.
129

-------
Gentile, J. H., and Scott, K. J. 1986. "The Application of a Hazard Assess-
ment Strategy to Sediment Testing: Issues and Case Study," Fate and Effects
of Sediment-Bound Chemicals in Aquatic Systems, K. L. Dickson, A. W. Maki, and
W. Brungs, eds., SETAC Special Publication pp 1-17.
Ivanovici, A. M. 1980a. "Adenylate Energy Charge in Pyyasus ebeninus in
Response to Salinity," Comparative Biochemistry and Physiology, Vol 66A,
PP 43-57.
	. 1980b. "Adenylate Energy Charge: An Evaluation of Applicability
to Assessment of Pollution Effects and Directions for Future Research,"
Rapport et Process-Verbaux des Reunions. Conseil International pour
l'Exploration de la Mer, Vol 179, pp 23-28.
Jenkins, K. D., and Brown, D. A. 1984. "Determining the Biological Signifi-
cance of Contaminant Bioaccumulation," Concepts in Marine Pollution Measure-
ments, H. H. White, ed., A Maryland Sea Grand Publication, University of
Maryland, College Park, Md., pp 355-375.
Johns, D. M., and Gutjahr-Gobell, R. 1988. "Bioenergetic Effects of Black
Rock Harbor Dredged Material on the Polychaete Nephtys i-nsisa: A Field Veri-
fication Study," Technical Report D-88-3, prepared by the US Environmental
Protection Agency, Narragansett, R. I., for the US Army Engineer Waterways
Experiment Station, Vicksburg, Miss.
Lake, J. L., Galloway, W. B., Hoffman, G., Nelson, W. G., and Scott, K. J.
1987. "Comparison of Field and Laboratory Bioaccumulation of Organic and
Inorganic Contaminants from Black Rock Harbor Dredged Material," Technical
Report D-87-6, prepared by US Environmental Research Laboratory, Narragansett,
R. I., for the US Army Engineer Waterways Experiment Station,
Vicksburg, Miss.
Lake, J., Hoffman, G., and Schimmel, S. 1985. "Bioaccumulation of Contami-
nants from Black Rock Harbor Dredged Material by Mussels and Polychaetes,"
Technical Report D-85-2, prepared by the US Environmental Protection Agency,
Environmental Research Laboratory, Narragansett, R. 1., for the US Army Engi-
neer Waterways Experiment Station, Vicksburg, Miss.
Lamprecht, W., and Trautschold, I. 1974. "Adenosine-5'-triphosphate: Deter-
mination with Hexokinase and Glucose-6-phosphate Dehydrogenase," Methods of
Enzymatic Analysis, H. V. Bergmeyer, ed., Second English edition, Verlag
Chemie Weinheim, Academic Press, New York, Vol 4, pp 2101-2110.
Lee, R. F., and Singer, S. C. 1980. "Detoxifying Enzymes System in Marine
Polychaetes: Increases in Activity After Exposure to Aromatic Hydrocarbons,"
Rapport et Process-Verbaux des Reunions. Conseil International pour
l'Exploration de la Mer, Vol 179, pp 29-32.
Lee, R. F,, Stolzenbach, J., Singer, S., and Tenore, K. R. 1981. "Effects of
Crude Oil on Growth and Mixed Function Oxygenase Activity in Polychaetes,
Nereis sp.," Biological Monitoring of Marine Pollutants, F. J. Vernberg,
A. Calabrese, F. P. Thurberg, and W. B. Vernberg, eds., Academic Press, New
York, pp 323-324.
Livingstone, D. R. 1985. "Responses of the Detoxification/Toxification
Enzyme Systems of Molluscs to Organic Pollutants and Xenobiotics," Marine
Pollution Bulletin, Vol 16, No. 4, pp 158-164.
130

-------
Montague, M. D., and Dawes, E. A. 1974. "The Survival of Peptococous
pvevotii in Relation to the Adenylate Energy Charge," Journal of General
Microbiology, Vol 80, pp 291-299.
Munns, W. R., Jr., Paul, J. F., Bierman, V. J., Jr., Davis, W. R., Galloway,
W. B., Hoffman, G. L., Rogerson, P. F., and Pruell, R. J. 1986. "Exposure
Assessment Component of the Field Verification Program: Data Presentation and
Synthesis," Contribution 751, US Environmental Protection Agency, Environmen-
tal Research Laboratory, Narragansett, R. I.
Nelson, W. G., Phelps, D. K., Galloway, W. B, Rogerson, P. F., and Pruell,
R. J. 1987. "Effects of Black Rock Harbor Dredged Material on the Scope for
Growth of the Blue Mussel, Mytilus edulis, After Laboratory and Field Expo-
sures," Technical Report D-87-7, prepared by US Environmental Research Labora-
tory, Narragansett, R. I, for the US Army Engineer Waterways Experiment
Station, Vicksburg, Miss.
Pesch, G. G., Mueller, C., Pesch, C. E., Malcolm, A. R., Rogerson, P. F.,
Munns, W. R., Jr., Gardner, G. R., Helthse, J., Lee, T. C., and Senecal, A. G.
1987. "Sister Chromatid Exchange in Marine Polychaetes Exposed to Black Rock
Harbor Sediment," Technical Report D-87-5, prepared by the US Environmental
Protection Agency, Narragansett, R. I., for the US Army Engineer Waterways
Experiment Station, Vicksburg, Miss.
Phelps, D., and Galloway, W. 1980. "A Report on the Coastal Environmental
Assessment Station (CEAS) Program," Rapport et Process-Verbaux des Reunions.
Conseil International pour 1'Exploration de la Mer, Vol 179, pp 76-81.
Rainer, S. F., Ivanovici, A. M., and Wadley, V. A. 1979. "The Effect of Re-
duced Salinity on Adenylate Energy Charge in Three Estuarine Molluscs," Marine
Biology, Vol 54, pp 91-99.
Ridge, J. W. 1972. "Hypoxia and the Energy Charge of the Cerebral Adenylate
Pool," Biochemical Journal, Vol 127, pp 351-355.
Roesijadi, G. 1982. "Uptake and Incorporation of Mercury into Mercury-
Binding Proteins of Gills of Mytilus edulis as a Function of Time," Marine
Biology, Vol 66, pp 151-157.
	. 1986. "Mercury-Binding Proteins from the Marine Mussel, Mytilus
edulisEnvironmental Health Perspectives, Vol 65, pp 45-48.
Rogerson, P. F., Schimmel, S. C., and Hoffman, G. L. 1985. "Chemical and Bio-
logical Characterization of Black Rock Harbor Dredged Material," Technical Re-
port D-85-9, US Army Engineer Waterways Experimental Station, Vicksburg, Miss.
SAS. 1985. SAS User's Guide; Statistics, Version 5 Edition, SAS Institute,
Inc., Gary, N. C.
Sinnett, J. C., and Davis, W. R. 1983. "A Programmable Turbidistat for Sus-
pended Particles in Laboratory Aquaria," Journal of Experimental Marine
Biology and Ecology, Vol 73, pp 167-174.
Skjoldal, H. R., and Bakke, T. 1978. "Relationship Between ATP and Energy
Charge During Lethal Metabolic Stress of the Marine Isopod Cirolana borealis,"
Journal of Biological Chemistry, Vol 253, pp 3355-3356.
Skjoldal, H. R., and Barkati, S. 1982. "ATP Content and Adenylate Energy
Charge of the Mussel Mytilus edulis During the Annual Reproductive Cycle in
Lindaspollene, West Norway," Marine Biology, Vol 70, pp 1-6.
131

-------
Snedecor, G. W., and Cochran, W. G. 1980. Statistical Methods, 7th ed., The
Iowa State University Press, Ames, Iowa.
US Environmental Protection Agency. 1979. "Methods for Chemical Analysis of
Water and Wastes," EPA-600/4-79-020, Environmental Monitoring and Support Lab-
oratory, Cincinnati, Ohio.
US Environmental Protection Agency/US Army Corps of Engineers. 1977. "The
Ecological Evaluation of Proposed Discharge of Dredged Material into Ocean
Waters; Implementation Manual for Section 103 of PL 92-532," Environmental
Effects Laboratory, US Army Engineer Waterways Experiment Station, Vicksburg,
Miss.
Vetter, R. D., and Hodson, R. E. 1984. "Metabolic Indicators of Sublethal
Stress: Changes in Adenine Nucleotides, Glycogen and Lipid," Concepts in
Marine Pollution Measurements, H. H. White, ed., A Maryland Sea Grant Publica-
tion, University of Maryland, College Park, Md., pp 471-498.
Wijsman, T. S. M. 1976. "Adenosine Phosphates and Energy Charge in Different
Tissues of Mytilus edulis Under Aerobic and Anaerobic Conditions," Journal of
Comparative Physiology, Vol 107, pp 129-140.
Zaroogian, G. E., Gentile, J. H., Heltshe, J. F., Johnson, M., and
Ivanovici, A. M. 1982. "Application of Adenine Nucleotide Measurements for
the Evaluation of Stress in Mytilus edulis and Crassostrea virginica
Comparative Biochemistry and Physiology, Vol 71B, pp 643-649.
Zaroogian, G. E., Pesch, C. E., Schauer, P., and Black, D. 1985. "Laboratory
Evaluation of Adenylate Energy Charge as a Test for Stress in Mytilus edulis
and Nephtys incisa Treated with Dredged Material," Technical Report D-85-3,
prepared by the US Environmental Protection Agency, Environmental Research
Laboratory, Narragansett, R. I., for the US Army Engineer Waterways Experiment
Station, Vicksburg, Miss.
132

-------
APPENDIX A: BLACK ROCK HARBOR SEDIMENT
PERCENTAGE CALCULATIONS

-------
Table A1
Percentage of Black Rock Harbor (BRH) Sediment in the Surflcial Sediments (0-2 cm)
and the Contaminants Used for the Percent Calculations
	Station	
Date	CNTR	200E	400E	1000E
Percentage of BRH Sediment
Jun
83
44.5
41.1
12.5
1.8
Jul
83
15.0
37.4
3.3
1.6
Sep
83
32.0
36.7
4.9
2.0
Dec
83
32.8
36.1
9.5
4.4
Mar
84
4.4
2.2
1.9
1.8
Jun
84
9.5
15.6
0.5
0.7
Sep
84
10.0
0.8
3.5
0.5
Oct
84
2.6
—
0.2
1.6
Dec
84
35.1
11.3
0.0
1.0
Oct
85
0.2
21.0
0.0
0.0
(Continued)

-------
Table A1 (Concluded)
Station
Date
Jun 83
Jul 83
Sep 83
Dec 83
Mar 84
Jun 84
Sep 84
Oct 84
Dec 84
Oct 85
CNTR
200E
400E
1000E
PAH+PCB+Cd+Cu+Cr
PAH+Cd+Cu+Cr
PAH+PCB+Cd+Cu+Cr
Cd+Cu+Cr
PAH+PCB+Cd+Cu+Cr
Cd+Cu+Cr
PAH+PCB+Cd+Cu+Cr
PAH+PCB
Cd+Cu+Cr
PCB
Contaminants Used*
PAH+PCB+Cd+Cu+Cr
PAH+PCB+Cd+Cu+Cr
PAH+PCB+Cd+Cu+Cr
Cd+Cu+Cr
PAH+PCB+Cd+Cu+Cr
Cd+Cu+Cr
PAH+PCB+Cd+Cu+Cr
Cd+Cu+Cr
PAH+PCB
PAH+PCB+Cd+Cu+Cr
PAH+PCB+Cd+Cu+Cr
PAH+PCB+Cd+Cu+Cr
Cd+Cu+Cr
PAH+PCB+Cd+Cu+Cr
Cd+Cu+Cr
PAH+PCB+Cu+Cr
PAH+PCB
Cu+Cr
PCB
Cd+Cu+Cr
PAH+PCB+Cd+Cu+Cr
PAH+PCB+Cd+Cu+Cr
Cd+Cu+Cr
PAH+PCB+Cu+Cr
Cu+Cr
PAH+PCB+Cu+Cr
PAH+PCB
Cu+Cr
PAH+PCB
* PAH = polynuclear aromatic hydrocarbons; PCB = polychlorinated biphenyls.

-------
Table A2
Phenanthrene Concentrations (ng/g Dry Weight) in
Surficial Sediments
Station
Date
CNTR
200E
400E
1000E
REFS
8/18/82
—
—
—
—
—
11/11/82
—
—
—
—
—
12/8/82
12/8/82
—
—
—
—
114
77
3/2/83
3/2/83
3/2/83
105
101
132
—
107
98
62
6/3/83
6/3/83
1,560
1,960
910
52
63
88
7/26/83
770
1,710
240
174
51
9/1/83
9/1/83
780
1,010
220
168
94
81
3/19/84
3/20/84
3/20/84
3/20/84
77
200
98
100
141
250
78
42
90
76
9/11/84
147
57
116
109
40
10/16/84
230
—
85
137
123
10/22/85
43
440
38
69
51
A5

-------
Table A3
178 Alkyl Homolog Concentrations (ng/g Dry Weight) in


Surficlal
Sediments





Station


Date
CNTR
200E
400E
1000E
REFS
8/18/82
—
—
—
—
—
11/11/82
—
—
—
—
—
12/8/82
— —
—
—
—
210
12/8/82
	
—
— —

172
3/2/83
250
210
260
	
188
3/2/83
—
—
—
— —
230
3/2/83
—
—
——
— ""
127
6/3/83

—
5,300
230
189
6/3/83
	
—
	
122
— —
7/26/83
9,700
—
1,500
412
131
9/1/83
5,200
—
1,480
613
186
9/1/83
—
—
	
	
189
3/19/84
1,330
590
560
600
103
3/20/84
—
—
590
260
170
3/20/84
—
—
—
—
185
3/20/84
1,200
—
—

— ~~
9/11/84
3,000
270
640
250
103
10/16/84
1,260
—
240
420
240
10/22/85
490
3,800
430
210
192
A6

-------
Table A4
Fluoranthene Concentrations (ng/g Dry Weight) in
Surficial Sediments
Station
Date
CNTR
200E
400E
1000E
REFS
8/18/82
—
—
—
—
—
11/11/82
—
—
—
—
—
12/8/82
12/8/82
—
—
—
—
280
200
3/2/83
3/2/83
3/2/83
300
260
340

270
230
148
6/3/83
6/3/83
2,300
2,300
1,240
142
161
220
7/26/83
1,940
2,600
570
400
140
9/1/83
9/1/83
1,370
2,800
560
380
220
210
3/19/84
3/20/84
3/20/84
3/20/84
290
510
330
330
360
600
210
124
230
185
9/11/84
650
166
410
250
108
10/16/84
580
—
240
320
300
10/22/85
172
1,770
142
196
189
A7

-------
Table A5
Benzo(a)pyrene Concentrations (ng/g Dry Weight) in
Surficial Sediments
Station
Date
CNTR
200E
400E
1000E
REFS
8/18/82
—
—
—
—
—
11/11/82
—
—
—
—
—
12/8/82
12/8/82
—
—
—
—
280
220
3/2/83
3/2/83
3/2/83
260
270
310
E
220
210
173
6/3/83
6/3/83
1,640
1,490
810
122
158
210
7/26/83
1,520
1,750
380
370
169
9/1/83
9/1/83
1,000
2,100
570
320
200
230
3/19/84
3/20/84
3/20/84
3/20/84
220
460
350
260
400
450
280
155
240
185
9/11/84
600
230
400
260
111
10/16/84
450
—
240
320
290
10/22/85
280
1,130
230
196
380
A8

-------


Table
A6



SUM PAH*
Concentrations
(ng/g Dry Weight)
in



Surficial
Sediments





Station


Date
CNTR
200E
400E
1000E
REFS
8/18/82
—
—
—
—
—
11/11/82
—
—
—
—
—
12/8/82



__
5,200
12/8/82
	
	
	
—
4,500
3/2/83
5,100
4,900
5,900
— _
4,400
3/2/83
—
—
—
— —
4,300
3/2/83
—
—
—
—
3,300
6/3/83
62,000
59,000
30,000
2,400
3,900
6/3/83
—
—
—
3,000
—
7/26/83
54,000
63,000
10,100
7,200
3,200
9/1/83
33,000
71,000
13,500
7,200
3,600
9/1/83
—
—
—
—
4,300
3/19/84
7,200
7,100
6,200
9,300
2,700
3/20/84
—
—
7,300
4,500
3,600
3/20/84
—
—

	
4,300
3/20/84
11,100
—
	
	
—
9/11/84
18,600
4,400
8,600
5,000
2,000
10/16/84
11,500
—
4,800
6,700
5,800
10/22/85
5,400
34,000
4,900
3,800
5,400
* PAH - polynuclear aromatic hydrocarbons.
A9

-------
Table A7
Centroid Statistic in Surficial Sediments
Station
Date
CNTR
200E
400E
1000E
REFS
8/18/82
—
—
—
—
—
11/11/82
—
—
—
—
—
12/8/82
12/8/82
—
—
—
—
249.7
252.0
3/2/83
3/2/83
3/2/83
247.6
248.9
247.7
—
247.4
248.0
252.1
6/3/83
6/3/83
238.7
234.1
235.2
241.4
250.3
248.3
7/26/83
234.7
232.6
234.4
247.3
252.5
9/1/83
9/1/83
239.7
238.6
244.7
244.3
245.4
250.3
3/19/84
3/20/84
3/20/84
3/20/84
237.0
242.9
245.1
241.1
243.5
244.5
245.3
251.0
243.7
251.5
9/11/84
240.8
249.2
244.1
247.5
247.2
10/16/84
240.4
—
248.4
247 .7
250.0
10/22/85
248.8
241.1
248.6
248.7
253.4
A10

-------


Table
A8



Ethvlan
Concentrations
: (ng/g Dry Weight)




in Surficial
Sediments





Station


Date
CNTR
200E
400E
1000E
REFS
8/18/82
—
—
—
—
—
11/11/82
—
—
—
—
—
12/8/82
__
	
	
	
0.0
12/8/82
	
	
	
—
0.0
3/2/83
0.0
0.0
0.0
	
0.0
3/2/83
—
—
—
—
0.0
3/2/83
—
—
—
—
0.0
6/3/83
340.0
370.0
163.0
5.0
0.0
6/3/83
—
—
—
0.0
—
7/26/83
0.0
950.0
90.0
35.0
0.0
9/1/83
210.0
670.0
30.0
15.0
0.0
9/1/83
—
—
—
—
0.0
3/19/84
74.0
50.0
36.0
31.0
0.0
3/20/84
—
—
12.0
0.0
0.0
3/20/84
—
—
—
—
0.0
3/20/84
23.0
—
—
—
—
9/11/84
96.0
14.0
64.0
3.0
0.0
10/16/84
12.0
—
2.0
7.0
0.0
10/22/85
8.0
820.0
4.0
5.0
0.0
All

-------
Table A9
PCB* (A1254) Concentrations (ng/g Dry Weight)
in Surficial Sediments
Station
Date
CNTR
200E
400E
1000E
REFS
8/18/82
—
—
73
—
59
11/11/82
—
—
30
—
26
12/8/82
—
—
—
—
48
3/2/83
3/2/83
3/2/83
77
75
98
—
65
67
60
6/3/83
6/3/83
1,730
1,650
890
79
45
59
7/26/83
180
1,830
240
117
28
9/1/83
1,190
2,200
340
200
59
3/19/84
3/20/84
270
181
250
162
96
26
9/11/84
440
113
183
66
27
10/16/84
181
—
84
162
77
10/22/85
72
1,550
37
48
29
* PCB - polychlorinated biphenyls.
A12

-------
Table A10
Cadmium Concentrations (ng/g Dry Weight)
in Surficial Sediments
Date


Station


CNTR
200E
400E
1000E
REFS
3/4/83
0.36
0.34
1.06
0.29
0.24
3/4/83
0.39
0.35
0.44
0.21
0.22
3/4/83
0.35
0.49
0.32
0.25
0.22
6/3/83
17.00
13.90
7.30
0.74
0.22
6/3/83
12.40
14.70
4.20
0.58
0.21
6/3/83
13.00
12.90
3.70
0.64
0.19
7/26/83
5.40
11.70
1.14
0.64
0.22
9/1/83
4.10
9.80
0.84
0.68
0.18
9/1/83
21.00
8.70
3.60
0.76
—
12/9/83
8.80
8.70
3.30
1.02
—
3/19/84
2.10
1.11
0.85
1.08
0.20
3/19/84
—
0.87
—
—
—
3/19/84
—
0.23
—
—
—
6/12/84
3.10
4.80
0.37
0.39
—
9/11/84
3.70
0.73
0.97
0.30
0.20
12/20/84
9.30
2.50
0.32
0.72
—
10/22/85
0.45
8.30
0.29
0.32
0.16
A13

-------
Table All
Chromium Concentrations (ng/g Dry Weight)
in Surficial Sediments
Station
Date
CNTR
200E
400E
1000E
REFS
3/4/83
56
39
59
59
48
3/4/83
53
57
43
58
52
3/4/83
45
56
56
60
54
6/3/83
870
680
340
69
49
6/3/83
780
740
191
72
48
6/3/83
800
600
155
74
48
7/26/83
120
519
69
66
44
9/1/83
310
600
106
79
56
9/1/83
680
380
160
79
—
12/9/83
520
660
117
126
—
3/19/84
100
52
54
86
47
3/19/84
—
140
—
—
—
3/19/84
—
40
—
—
—
6/12/84
138
210
41
52
—
9/11/84
153
41
128
55
44
12/20/84
550
175
47
88
—
10/22/85
54
430
57
59
40
AH

-------
Table A12
Copper Concentrations (ng/g Dry Weight)
in Surficial Sediments
Date


Station


CNTR
200E
400E
1000E
REFS
3/4/83
67
57
67
70
55
3/4/83
62
69
63
68
57
3/4/83
63
67
64
69
58
6/3/83
1,640
1,380
680
99
48
6/3/83
1,300
1,420
360
102
51
6/3/83
1,330
1,240
303
106
56
7/26/83
450
1,230
185
106
49
9/1/83
560
1,070
134
103
47
9/1/83
1,890
910
510
122
—
12/9/83
910
950
370
177
—
3/19/84
200
111
143
123
53
3/19/84
—
107
—
—
—
3/19/84
—
114
—
—
—
6/12/84
350
530
89
83
—
9/11/84
430
86
156
73
48
12/20/84
1,000
500
52
131
—
10/22/85
92
910
75
72
46
A15

-------
Table A13
Iron Concentrations (ng/g Dry Weight)
in Surficial Sediments



Station


Date
CNTR
200E
400E
1000E
REFS
3/4/83
3/4/83
3/4/83
21,000
20,000
18,400
17,100
22,000
21,000
22,000
18,900
21,000
23,000
23,000
23,000
19,700
21,000
22,000
6/3/83
6/3/83
6/3/83
17,100
19,300
17,900
19,200
19,000
18,700
23,000
22,000
23,000
21,000
21,000
22,000
21,000
19,000
21,000
7/26/83
15,200
16,700
21,000
16,800
21,000
9/1/83
9/1/83
15,100
26,000
19,300
15,100
21,000
18,400
16,400
19,700
12/9/83
16,500
21,000
19,600
17,500
—
3/19/84
3/19/84
3/19/84
5,800
17,300
16,600
15,600
20,000
18,700
21,000
6/12/84
6,500
17,100
19,800
15,600
—
9/11/84
12,600
17,400
18,400
18,200
21,000
12/20/84
18,100
17,300
17,400
18,000
—
10/22/85
9,900
17,200
18,100
18,900
17,000
A16

-------
APPENDIX B: CHEMICAL FORMULAS AND FIELD WORM RESIDUE CONCENTRATIONS

-------
Table B1
Chemical Contaminants Selected for Measurement
in Both Field and Laboratory Studies
Chlorinated hydrocarbon pesticides
Polychlorinated biphenyls
Ethylan
Aromatic hydrocarbons > molecular weight 166:

Molecular
Compound Class
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
____	(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.
B3

-------
Table Bl (Concluded)
Molecular
Compound Class		Weight
Benzofluoranthenes	252
Benzo(e)pyrene	252
Benzo(a)pyrene	252
Perylene	252
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
Dibenzothiopehen	184
C-l* Dibenzothiophene	198
C-2* Dibenzothiophene	212
C-3* Dibenzothiophene	226
C-4* Dibenzothiophene	240
Metals
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.
B4

-------
Table B2
Complete Formulae for Calculating all SUM and CENT Variables
PSUM
= POS166
+
POS178
+
POS202
+
POS228
+
POS252
+
POS276
+
POS278
+

POS300
+
P0S302











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
+
POS302
PCENT = [POS166*166 + POS178*178 + POS202*202 + POS228*228 + POS252*252 +
POS276*276 + POS278*278 + POS300*300 + POS302*302]/PSUM
HCENT = [H1C166*180 + H2C166*194 + H3C166*208 + H4C166*222 + H1C178*192 +
H2C178*206 + H3C178*22Q + H4C178*234 + H1C202*216 + H2C202*230 +
H3C202*244 + H4C202*258 + H1C228*242 + H2C228*256 + H3C228*270 +
H4C228*284 + H1C252*266 + H2C252*280 + H3C252*294 + H4C252*308]/HSUM
CENT = [POS166*166 + H1C166*180 + H2C166*194 + H3C166*208 + H4C166*222 +
POS178*178 + H1C178*192 + H2C178*206 + H3C178*220 + H4C178*234 +
POS202*202 + H1C202*216 + H2C202*230 + H3C202*244 + H4C202*258 +
POS228*228 + H1C228*242 + H2C228*256 + H3C228*270 + H4C228*284 +
POS252*252 + H1C252*266 + H2C252*280 + H3C252*294 + H4C252*308 +
POS276*276 + POS278*278 + POS300*300 + 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.
B5

-------
Table B3
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	117	98	38
Sum of 178 alkyl	580	310	310	290
homologs
Fluoranthene	161	102	90	82
Benzo(a)pyrene	37	20	34	25
Ethylan	5	3	5	10
PCB as A1254	380	270	400	440
SUM of PAHs	2,600	1,520	1,650	1,380
CENTROID of PAHs	218	219	225	228
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	1,400	530	340
* 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.
B6

-------
Table B4
Tissue Residue Concentrations in Mussels from the T + 0
Field Collection in CLIS (24 May 83)*
Station**
Chemical Compound	1000E	REFS
Phenanthrene	43	16
Sum of 178 alkyl	1,440	290
homologs
Fluoranthene	161	52
Benzo(a)pyrene	100	18
Ethylan	102	9
PCB as A1254	1,080	500
SUM of PAHs	5,400	1,290
CENTROID of PAHs	230	232
Copper	16.5	10.9
Cadmium	2.0	2.3
Chromium	2.6	1.5
Iron	420	330
* 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.
** The CNTR station was not deployed because of the dumping operation and the
400E station was lost.
B7

-------
Table B5
Tissue Residue Concentrations in Mussels from the T + 2
Field Collection in CLIS (07 Jun 83)*
Station**
Chemical Compound	400E	1000E	REFS
Phenanthrene	69	41	13
Sum of 178 alkyl	1,900	970	540
homologs
Fluoranthene	290	126	72
Benzo(a)pyrene	210	118	51
Ethylan	71	39	17
PCB as A1254	1,440	1,020	630
SUM of PAHs	8,700	4,700	2,500
CENTROID of PAHs	232	234	233
Copper	16.9	15.6	10.8
Cadmium	2.3	2.3	1.9
Chromium	3.0	3.0	2.0
Iron	510	560	560
* 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.
** The CNTR station was not deployed because of the disposal operation.
B8

-------
Table B6
Tissue Residue Concentrations in Mussels from the T + 8
Field Collection in CLIS (10 Jul 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
11
350
45
40
22
700
1,870
234
10.1
1.9
1.4
340
400E
14
340
46
50
20
740
2,100
236
9.6
2.0
1.4
370
1000E
9
193
31
18
7
620
1,020
231
11.5
1.3
3.2
820
REFS
7
105
23
20
1
480
760
240
4.4
0.9
0.8
240
* 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.
B9

-------
Table B7
Tissue Residue Concentrations in Mussels from the T + 12
Field Collection in CLIS (10 Aug 83)*
Station
Chemical Compound	CNTR	400E	1000E	REFS
Phenanthrene	17	10	9	8
Sum of 178 alkyl	250	160	96	65
homologs
Fluoranthene	41	28	20	15
Benzo(a)pyrene	41	17	16	13
Ethylan	9	8	3	1
PCB as A1254	640	660	550	570
SUM of PAHs	1,600	940	710	530
CENTROID of PAHs	237	236	239	240
Copper	5.3	5.6	7.5	5.8
Cadmium	0.9	0.9	1.2	1.1
Chromium	1.0	0.7	1.6	0.7
Iron	164	167	450	177
* 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.
B10

-------
Table B8
Tissue Residue Concentrations in Mussels from the T + 15
Field Collection in CLIS (06 Sep 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
13
370
57
53
10
870
2,100
236
7.7
1.0
1.2
260
400E
9
230
38
45
6
630
1,540
239
6.0
1.1
0.9
179
1000E
10
210
33
28
4
640
1,240
237
8.0
1.1
1.1
290
REFS
6
43
14
7
1
550
350
238
5.8
0.9
0.9
260
* 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.
Bll

-------
Table B9
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
12
132
33
24
2
540
1,000
240
22.1
5.1
2.3
440
400E
11
101
25
9
2
680
670
234
16.3
4.4
2.6
540
1000E
11
88
22
17
1
570
700
239
15.1
4.8
2.2
420
REFS
10
46
16
9
0
420
400
238
16.4
5.0
2.2
480
* 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.
B12

-------
Table BIO
Tissue Residue Concentrations in Mussels from the T + 27
Field Collection in CLIS (29 Nov 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
4Q0E
18
230
68
39
3
540
1,820
240
16.4
3.2
2.5
570
1000E
10
117
36
32
1
380
1,150
244
21.0
3.4
3.6
920
REFS
8
86
37
19
0
450
860
240
23.3
3.6
3.3
920
* 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.
** The CNTR station was missing at the time of collection.
B13

-------
Table Bll
Tissue Residue Concentrations in Mussels from the T + 43
Field Collection in CL1S (20 Mar 84)*
Station**
Chemical Compound
Phenanthrene
Sum of 178 alkyl
homologs
Fluoranthene
Benzo(a)pyrene
Ethylan
PCB as A1254
SUM of PAHs
CENTROID of PAHs
CNTR
6
94
28
8
2
350
510
229
400E
18
78
26
4
1
330
460
230
REFS
17
70
24
7
1
280
450
231
* Metals were not measured for these samples. Units are nanograms per gram
dry weight for the organic compounds and the statistic SUM, and molecular
weight for the statistic CENTROID.
** The 1000E station was missing at the time of collection.
B14

-------
Table B12
Tissue Residue Concentrations in Mussels from the T + 55
Field Collection in CLIS (05 Jun 84)*
Station**
Chemical Compound	400E	1000E	REFS
Phenanthrene	6	6	4
Sum of 178 alkyl	89	91	54
homologs
Fluoranthene	25	31	18
Benzo(a)pyrene	6	7	6
Ethylan	0	10
PCB as A1254	540	490	550
SUM of PAHs	520	550	370
CENTROID of PAHs	234	235	236
* Metals were not measured for these samples. Units are nanograms per gram
dry weight for the organic compounds and the statistic SUM, and molecular
weight for the statistic CENTROID.
** The CNTR station was missing at the time of collection.
B15

-------
Table B13
Tissue Residue Concentrations in Mussels from the T + 116
Field Collection in CLIS (13 Aug 85)*
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
3
79
18
18
1
310
700
242
8.5
1.5
1.2
290
400E
6
124
27
40
2
350
1,270
243
7.5
1.3
1.1
260
1000E
3
80
24
22
1
450
810
241
6.9
1.3
0.9
220
REFS
3
58
19
17
0
440
620
244
7.6
1.3
0.9
220
* 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.
B16

-------