v
FIELD VERIFICATION PROGRAM
(AQUATIC DISPOSAL)
TECHNICAL REPORT D-85-9
CHEMICAL AND BIOLOGICAL CHARACTERIZATION
OF BLACK ROCK HARBOR DREDGED MATERIAL
by
Peter F. Rogerson, Steven C. Schimmel
Gerald Hoffman
US Environmental Protection Agency
Environmental Research Laboratory
Narragansett, Rhode Island 02882
September 1985
Final Report
Approved For Public Release; Distribution Unlimited
Prepared for DEPARTMENT OF THE ARMY
US Army Corps of Engineers
Washington, DC 20314-1000
and US Environmental Protection Agency
Washington, DC 20460
Monitored by Environmental Laboratory
US Army Engineer Waterways Experiment Station
PO Box 631, Vicksburg, Mississippi 39180-0631
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Destroy this report when no longer needed. Do not return
it to the originator.
The findings in this report are not to be construed as an official
Department of the Army position unless so designated
by other authorized documents.
The contents of this report are not to be used for
advertising, publication, or promotional purposes.
Citation of trade names does not constitute an
official endorsement or approval of the use of
such commercial products.
The D-series of reports includes publications of the
Environmental Effects of Dredging Programs:
Dredging Operations Technical Support.
Long-Term Effects of Dredging Operations
Interagency Field Verification of Methodologies for
Evaluating Dredged Material Disposal Alternatives
(Field Verification Program)
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SUBJECT: Transmittal of Field Verification Program Technical Report Entitled
"Chemical and Biological Characterization of Black Rock Harbor
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
Predictive 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 (EKLN) 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 inter-
pretive approaches applicable to evaluation of many dredging and disposal
operations. 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 a generic predictive hazard-assessment research
strategy applicable to all wastes disposed in the aquatic environment. There-
fore, the ERLN initiated exposure-assessment studies at the aquatic disposal
site. The Corps-sponsored studies on environmental consequences of aquatic
disposal will provide the effects assessment necessary to complement the EPA-
sponsored exposure assessment, thereby allowing ERLN to develop and apply a
hazard-assessment strategy. While not part of the Corps-funded FVP, the EPA
exposure assessment studies will complement the Corps' work, and together the
Corps and the EPA studies will satisfy the needs of both agencies.
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SUBJECT: Transmittal of Field Verification Program Technical Report Entitled
"Chemical and Biological Characterization of Black Rock Harbor
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
Dredging Programs at WES. Studies of the effects of upland disposal and
wetland creation are being conducted by WES and studies of aquatic disposal
are being carried out by the ERLN, applying techniques worked out at the
laboratory for evaluating sublethal effects of contaminants on aquatic organ-
isms. These studies are funded by the Corps while salary, support facilities,
etc., are 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 con-
siderable. The studies conducted under this program are scientific in nature
and will be 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 implementa-
tion, and perhaps regulations themselves, will be based. However, the docu-
ments 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.
Choromokos, Jr., Ph.D., P.E.
Director, Research and Development
U. S. Army Corps of Engineers
Bernard D. Goldstein, M.D.
Assistant Administrator for
Research and Development
U. S. Environmental Protection
Agency
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Unclassified
SECURITY CLASSIFICATION OF THIS PAGE (When Data Entered)
REPORT DOCUMENTATION PAGE
READ INSTRUCTIONS
BEFORE COMPLETING FORM
1. REPORT NUMBER
Technical Report D-85-9
2. GOVT ACCESSION NO
3. RECIPIENT'S CATALOG NUMBER
4. TITLE (and Subtitle)
CHEMICAL AND BIOLOGICAL CHARACTERIZATION OF
BLACK ROCK HARBOR DREDGED MATERIAL
5. TYPE OF REPORT & PERIOD COVERED
Final report
6. PERFORMING ORG. REPORT NUMBER
7. AUTHORO)
Peter F. Rogerson, Steven C. Schimmel,
Gerald Hoffman
8. CONTRACT OR GRANT NUMBERfa.)
9. PERFORMING ORGANIZATION NAME AND ADDRESS
US Environmental Protection Agency
Environmental Research Laboratory
Narragansett. Rhode Island 02882
10. PROGRAM ELEMENT, PROJECT, TASK
AREA & WORK UNIT NUMBERS
Field Verification Program
(Aquatic Disposal)
It. CONTROLLING OFFICE NAME AND ADDRESS
DEPARTMENT OF THE ARMY, US Army Corps of Engineers,
Washington, DC 20314-1000 and US Environmental
Protection Agency, Washington. DC 20460
12. REPORT DATE
September 1985
13. NUMBER OF PAGES
123
14. MONITORING AGENCY NAME & AODRESSf/f different from Controlling O(llce)
US Army Engineer Waterways Experiment Station
Environmental Laboratory
PO Box 631, Vicksburg, Mississippi 39180-0631
15. SECURITY CLASS, (of thlm report)
Unclassified
DECLASSIFICATION/DOWN GRADING
SCHEDULE
16. DISTRIBUTION STATEMENT (of thfa Report)
Approved for public release; distribution unlimited.
17. DISTRIBUTION STATEMENT (of Ota abstract entered In Block 30, It different from Report)
IB. SUPPLEMENTARY NOTES
Available from National Technical Information Service, 5285 Port Royal Road,
Springfield, Virginia 22161. Appendix B was prepared on microfiche and is
enclosed in an envelope attached to the back cover of this report.
19. KEY WORDS (Continue on revere* tide II neceuary and Identify by block number)
Dredging—Connecticut—Black Rock Harbor (LC)
Black Rock Harbor (Conn.) (LC)
Marine sediments—Analysis (LC)
Dredged material (WES)
Aquatic biology (LC)
20. ABSTRACT fCantBaum «ra rererea ttit* It rocwuir and Identity by block number)
Black Rock Harbor, Bridgeport, Conn., dredged material contained substan-
tial concentrations of both organic and inorganic contaminants, many of which
were shown to be biologically available to the blue mussel, Mytilus edulis, in
a laboratory bioassay. Tissue,PCB concentrations were 44% of the concentration
found in the sediment (6800 ng/g), while tissue concentrations of parent poly-
nuclear hydrocarbons were 28% of sediment concentrations that ranged up to
9800 ng/g. Also present in the sediment were Cu, Cr, Zn, Pb, Ni, Cd, and
. (Continued)
DD,
FORM
JAN 73
1473
EDITION OF t NOV SS IS OBSOLETE
Unclassified
SECURITY CLASSIFICATION OF THIS PA!>E (Whm Date Entered)
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Unclassified
SECURITY CLASSIFICATION OF THIS PAGEfHTien D»t* Entered;
20. ABSTRACT (Continued).
Hg at 2380, 1430, 1200, 380, 140, 23, and 1.7 Pg/g, respectively. Of these,
Cu, Cr, Pb, Ni, and Cd accumulated in the mussels.
In acute solid phase toxicity tests, the sediment was lethal to only one
of the eleven species tested, Aropelisca abdita, although behavioral changes
were observed in two additional species, both infaunal species. No effect was
noted with epibenthic or water column species in either solid phase or in
combination with suspended particulate phase.
This investigation is the first phase in developing field-verified bio-
assessment evaluations for the Corps of Engineers and the US Environmental,
Protection Agency regulatory program for dredged material disposal. This
report is not suitable for regulatory purposes; however, appropriate assessment
methodologies that are field verified will be available at the conclusion of
this program.
Unclassified
SECURITY CLASSIFICATION OF THIS PAGEfHTi.n DM* Enlar«f>
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PREFACE
This report describes work performed by the U.S. Environmental
Protection Agency (EPA) Environmental Research Laboratory, Narragansett,
R.I. (ERLN), as part of the Interagency Field Verification of Testing
and Predictive Methodologies for Dredged Material Disposal Alternatives
Program (Field Verification Program (FVP)). This Program is sponsored
by the Office, Chief of Engineers (OCE), and is assigned to the U.S. Army
Engineer Waterways Experiment Station (WES), under the purview of the
Environmental Laboratory's (EL) Environmental Effects of Dredging Pro-
grams (EEDP). The OCE Technical Monitors for FVP were Drs. William L.
Klesch and Robert J. Pierce. The objective of this interagency program
is to field verify existing predictive techniques for evaluating the
environmental consequences of dredged material disposal under aquatic,
wetland, and upland conditions. The aquatic portion of the FVP study
is being conducted by ERLN, with the wetland and upland portions con-
ducted by WES.
The principal ERLN investigators for this aquatic study were
Drs. Peter Rogerson and Gerald Hoffman, Analytical Chemists, and Mr.
Steven Schimmel, Aquatic Toxicologist. Laboratory exposure system
design was coordinated by Mr. Jay Sinnett, Ms. Dianne Black, Dr. Wayne
Davis, and Mr. John Sewall. Organic chemical sample preparation and
analysis were conducted by Ms. Sharon Pavignano, Mr. Larry LeBlanc,
Ms. Adria Elskus, Mr. Robert Bowen, and Mr. Curt Norwood under the
supervision of Drs. Rogerson and James Lake. Inorganic chemical prepa-
ration and analysis was conducted under the supervision of Dr. Gerald
Hoffman, and assisted by Mr. Frank Osterman, Mr. Warren Boothman, and
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Mr. Dennis Migneault. Biological testing was conducted by Dr. John
Scott, Dr. Paul Schauer, Mr. Walter Berry, Ms. Suzanne Lussier Gentile,
Ms. Michele Redmond, Ms. Melissa Hughes, Dr. Chris Deacutis, Dr. Grace
MacPhee, and Ms. Ann Kuhn. Data management and analysis was conducted
by Mr. Jeffery Rosen.
The EPA Technical Director for the FVP was Dr. John H. Gentile; the
Technical Coordinator was Mr. Walter Galloway; and the Project Manager
was Mr. Allan Beck.
The study was conducted under the direct management of Drs. Thomas M.
Dillon and Richard K. Peddicord of the Contaminant Mobility and Criteria
Group (CMCG), Ecosystem Research and Simulation Division (ERSD), EL;
and under the general management of Dr. Charles R. Lee, Chief, CMCG,
Mr. Donald L. Robey, Chief, ERSD, and Dr. John Harrison, Chief, EL. The
FVP Coordinator was Mr. Robert L. Lazor, and the Program Managers were
Mr. Charles C. Calhoun, Jr., and Dr. Robert M. Engler. The report was
edited by Ms. Jamie W. Leach of the WES Publications and Graphic Arts
Division.
During preparation of this report, COL Tilford C. Creel, CE, and
COL Robert C. Lee, CE, were Commanders and Directors of WES and Mr. F. R.
Brown was Technical Director. At the time of publication, COL Allen F.
Grum, USA, was Director and Dr. Robert W. Whalin was Technical Director.
This report should be cited as follows:
Rogerson, P.F., Schimmel, S.C., and Hoffman, G. 1985.
"Chemical and Biological Characterization of Black Rock
Harbor Dredged Material," Technical Report D-85-9, prepared
by US Environmental Protection Agency, Narragansett, R.I.,
for the US Army Engineer Waterways Experiment Station,
Vicksburg, Miss.
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CONTENTS
Page
PREFACE 1
LIST OF FIGURES 4
LIST OF TABLES , 6
PART I: INTRODUCTION 8
Background 8
Purpose * 8
Scope 9
PART II: GENERAL METHODS AND MATERIALS 12
Sediment Collection and Preservation 13
Sediment Dosing System 16
PART III: CHEMICAL CHARACTERIZATION METHODS AND MATERIALS 20
Contaminant Uptake Test 20
Sample Preparation 25
Sample Analysis 32
PART IV: BIOLOGICAL CHARACTERIZATION METHODS AND MATERIALS 36
Overview 36
Annelids 40
Molluscs 43
Arthropods 47
Fishes 52
PART V: RESULTS AND DISCUSSION . 61
Chemical Characterization 61
Biological Characterization 90
PART VI: CONCLUSIONS AND RECOMMENDATIONS 104
REFERENCES 108
APPENDIX A: CHEMICAL DATA Al
APPENDIX B: ACUTE TOXICITY DATA SHEETS* Bl
* Appendix B was prepared on microfiche and is enclosed in an envelope
attached to the back cover of this report.
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LIST OF FIGURES
No. Page
1 Principal components of sediment characterization 13
2 Central Long Island Sound disposal site and South
reference site 14
3 Black Rock Harbor, Connecticut, source of dredged material... 16
4 Sediment dosing system with chilled water bath and argon
gas supply 18
5 Suspended sediment feedback control loop and strip
chart recorder 19
6 Blue mussel (Mytilus edulis) contaminant uptake system 21
7 Suspended sediment dilution system, distribution chamber,
and exposure chambers used for acute toxicity .tests 39
8 Distribution and exposure chambers used for solid phase
and suspended particulate phase exposure of Yoldia limatula,
Mulinia lateralis, and Ampelisca abdita 46
9 Total ion current profiles of the 28-day exposed mussels
analyzed by GC-MS with a 4°/min (50-330) temperature
programming rate 62
10 Total ion current profiles of the 28-day control mussels
analyzed by GC-MS with a 4°/min (50-330) temperature
programming rate 63
11 Total ion current profiles of the Black Rock Harbor
FVP reference sediment analyzed by GC-MS with a
4°/min (50-330) temperature programming rate 72
12 Concentration of PAH compounds in Black Rock Harbor sediment
and exposed and control mussels 75
13 Concentration of sum of C-l through C-4 alkyl homologs
of PAH's measured in Black Rock Harbor sediment
and control and exposed mussels 76
14 Distribution of Cr versus Fe in mussels from the
Black Rock Harbor exposure • • 84
15 Distribution of Pb versus Fe in mussels from the
Black Rock Harbor exposure 85
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LIST OF FIGURES (Cont'd)
No. Page
16 Distribution of Cd versus Fe in mussels from the
Black Rock Harbor exposure 85
17 Distribution of Zn versus Fe in mussels from the
Black Rock Harbor exposure 86
18 Distribution of Cu versus Fe in mussels from the
Black Rock Harbor exposure 87
19 Distribution of As versus Fe in mussels from the
Black Rock Harbor exposure 88
20 Distribution of Mn versus Fe in mussels from the
Black Rock Harbor exposure 88
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LIST OF TABLES
No. page
1 Summary of Experimental CondiCions for the Contaminant
Uptake Study with My til us edulis 25
2 Test Species, Exposure Conditions, and Food Used for Solid
Phase Biological Assays for Sediment Characterization 37
3 Test Species, Exposure Conditions, and Food for Suspended
Particulate Phase Assays for Sediment Characterization 44
.;•}
4 Single Replicate PCB Concentrations as Aroclor®1254 in
ng/g Dry Weight Including Chlorine Number Distributions
by Mass Spectrometry 64
5 Parent Polynuclear Aromatic Hydrocarbons Found in the
Exposed Mussels and Black Rock Harbor Sediment 66
6 Mean Concentrations + Standard Deviation of Parent PAH
Compounds Found in Exposed and Control Mussels and in
Black Rock Harbor Sediment in ng/g Dry Weight 68
7 Mean Concentrations + Standard Deviation of the Sum of C-l
Through C-4 Alkyl Homologs of PAH's Found in Exposed and
Control Mussels and in Black Rock Harbor Sediment
Quantitated as Each Parent PAH 69
8 Distribution of Five Silicone-like Compounds in Exposed
and Control Mussels and Black Rock Harbor Sediment Measured
as GC-MS Area Counts/Gram Dry Weight 70
9 Organic Contaminants in Black Rock Harbor Sediment,
ng/g Dry Weight, One Replicate 73
10 Distribution of Trace Elements in g/g Dry Weight
in Exposed and control Mussels '. 78
11 Average Metal Concentrations in Black Rock Harbor
Barrel #00 and Barrel //LL Sediment Samples 80
12 Ratios of Trace Metal Accumulations in Exposed
and Control Mussels 82
13 Toxicity of Solid Phase Black Rock Harbor (Connecticut)
Dredged Material to 11 Species of Marine
Invertebrates and Fishes 91
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LIST OF TABLES (Cont'd)
No. Page
14 Percent of Yoldia not Burrowed into Sediment over Time for
Solid Phase Test No. 1 and Suspended Phase Test No. 1 93
15 Summary of Response Percent Mortality of Ampelisca abdita
after 96-hr Exposure in Solid Phase Tests with
Black Rock Harbor Sediment 96
16 Toxicity of Black Rock Harbor Dredged Material,
as Suspended Sediment, to Ten Species of Marine
Invertebrates and Fishes 100
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CHEMICAL AND BIOLOGICAL CHARACTERIZATION OF
BLACK ROCK HARBOR DREDGED MATERIAL
PART I: INTRODUCTION
Background
1. The U.S. Army Corps of Engineers (CE) and the U.S. Environ-
mental Protection Agency (EPA) are jointly conducting a comprehensive
Field Verification Program (FVP) to evaluate the risk associated with
various disposal options for dredged material. The approach being
used in the FVP is to evaluate and field validate assessment methodol-
ogies for predicting the environmental impacts of dredged material
disposal in aquatic, upland, and wetland environments. The research,
evaluation, and field verification of the upland and wetland disposal
options is being conducted by the Environmental Laboratory, U.S. Army
Engineer Waterways Experiment Station (WES), Vicksburg, Miss. The
application and field verification of predictive methodologies for the
aquatic disposal option is being conducted by the EPA Environmental
Research Laboratory (ERL-N), Narragansett, R.I.
Purpose
2. The aquatic disposal alternative of the FVP is being used
as a site-specific case study to evaluate a hazard assessment research
strategy. Hazard assessment in terms of this study is a process by
which data on exposure and effects are assembled and interpreted to
determine the potential for harm to the aquatic environment that could
8
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result from the ocean disposal of a particular material. To measure
this hazard, information on the duration and intensity of exposure
(exposure assessment) of organisms and the concentrations of contami-
nants in the materials disposed at the site (predicted environmental
concentration) is coupled with concentrations of the contaminants
determined to be toxic to individual species, populations, and commu-
nities in laboratory toxicity studies (effects assessment). When
properly synthesized, these data provide an estimate of the probability
(risk) of unacceptably altering the aquatic environment as a result of
the disposal of the materials. The verification of hazard assessment
is comprised of two components: verification of an individual method
or protocol between the lab and field, and verification of the predic-
tion of risk to the aquatic environment. Within this context, hazard
assessment contains parallel predictive laboratory and field verifica-
tion components. The achievement of the goal of hazard assessment
requires the development and verification of assessment protocols for
defining exposure and effects.
Scope
3. The first research component in the aquatic portion of the FVP
is sediment characterization, which includes chemical and biological
characterization of the dredged material.
Chemical characterization
4. Chemical characterization is focused on determining what
chemical contaminants are present in the dredged material and, of
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these, which bioaccumulate and constitute a potential threat to man
and the ecosystem. The approach taken in chemical characterization is
to allow the environment, 'in the form of an organism (Mytilus edulis),
to indicate contaminants of biological importance from the dredged mate-
rial. These contaminant profiles can then be compared with chemical
profiles both from the bulk sediment analysis and a more detailed
sediment chemistry analysis of organic contaminants. This approach
has several advantages over chemical screening for preselected compounds
(e.g., 129 priority pollutants):
a_. Chemical screening of preselected chemicals generally
results in a large number of chemicals being classified
as nondetectable.
b. Preselection reduces the number of contaminants examined.
Eliminating preselection and allowing an organism to bioaccumulate con-
taminants increases the probability of detection, reduces the risk of
a biologically important contaminant not being detected, and focuses
the research and monitoring efforts on those contaminants known to
bioaccumulate. The one element of uncertainty in this approach is that
there may be contaminants which go undetected that are biologically
active but do not bioaccumulate. The risk of this occurring, however,
is considered to be relatively negligible.
Biological characterization
5. Biological characterization focuses on "worst-case" toxicological
evaluation of dredged material. As such, the exposure regimes do not
necessarily reflect actual field conditions. The approach selected is to
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adapt existing toxicological protocols for use with solid and suspended
participate phase flow-through tests for both indigenous and "surrogate"
test species. Each acute toxicity test will be evaluated for its appli-
cability and sensitivity for detecting and measuring dredged material
effects. Determinations of test and method variability and reproduci-
bility will be made where appropriate. Finally, results from these
tests will be used to help design exposure conditions for future sub-
lethal biological effects tests.
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PART II: GENERAL METHODS AND MATERIALS
6. The principal components of this study and their interrela-
tionships are schematically represented in Figure 1. The methods for
sediment characterization begin with the suspended sediment (SS) dosing
system. This system is designed to maintain reservoirs of reference
sediment and dredged material under defined anoxic conditions and to
quantitatively deliver them through recirculating loops to test systems.
7. In studies on chemical characterization, known quantities of
suspended dredged material were delivered to the contaminant uptake
system containing the bivalve mollusc Mytilus edulis, where chemical
analyses were conducted for contaminants within the tissues, dissolved
in the water, and in the particulate phase from the sediment dosing
system. These analyses were compared with sediment chemical analyses
to determine which of the many sediment contaminants were bio-
accumulated.
8. The suspended sediment dosing system also interfaced with a
series of experimental components in the studies for biological charac-
terization. Sediment suspensions were first quantitatively delivered
to a controlled dilution system. Here, diluted sediment suspensions
were maintained using a transmissometer-microprocessor feedback system.
These suspensions then flowed into multiported distribution systems
which fed a constant, prescribed dilution of suspension to the suspended
sediment toxicity test chambers. Toxicological information was produced
for polychaetous annelids, bivalve molluscs, arthropods (crustaceans),
and fishes.
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SUSPENDED SEDIMENT (SS)
DOSING SYSTEM
CE
Bulk Sediment
Analysis
CHEMICAL
'CHARACTERIZATION'
I
BIOLOGICAL
CHARACTERIZATION
SS-Controlled
Dilution System
Dissolved
Contaminants
Particulate
Contaminants
SS-Distribution
System
SS-Toxicity
Test System
Figure 1. Principal components of sediment characterization
Sediment Collection and Preservation
Reference sediment
9. Reference sediment (REF) for the FVP sediment characteriza-
tion studies was collected from the South Reference site (41°7.95"N
and 72°52.7"W), which is approximately 700 m south of the southern
perimeter of the Central Long Island Sound (CLIS) disposal site (Figure
2). Reference sediment was collected with a Smith-Maclntyre grab
sampler (0,1 m^) in both August and December 1982. Sediment collected
on each date was returned to the laboratory, press seived (wet) within
48 hr through a 2-mm mesh stainless steel screen, homogenized, and
stored at 48C until used for experimental purposes. Sediment from the
13
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August collection was stored in 32 cm x 61 cm x 25 cm (38 L) polypro-
pylene containers. Sediment in each container was allowed to reach
room temperature and rehomogenized (mixed) prior to use.
BLACK ROCK
HARBORJ
FVP
DISPOSAL
SITE
SOUTH REFERENCE
• SITE
Figure 2. Central Long Island Sound disposal site
and South reference site
Sediment from the December collection was stored in 3.8-L glass jars
with polypropylene lids. Each jar of material was coded with a collec-
tion date, batch number, bottle number, and the name of the person to
whom the material was assigned.
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Black Rock Harbor sediment
10. Black Rock Harbor is locatd in Bridgeport, Conn. (Figure 3),
with the approximate coordinates of 73°13"W and 41°9" N. The study
reach begins 400 m south of the fork in Cedar Creek and extends seaward
for approximately 1700 m. Black Rock Harbor (BRH) bottom sediments
were collected at 25 locations within the study area using a O.l-m^
gravity box corer to a depth of 1.21 m and placed in 210-L barrels and
transported in a refrigerated truck (at 4°C) to WES. The contents of
the 25 barrels were emptied into a nitrogen-purged cement mixer and
homogenized. The homogenized sediment was then redistributed to the
25 barrels and aliquots were taken from each for sediment chemical
analysis. Thirteen barrels were then transported to ERL-N in a refrig-
erated truck and stored at 4°C. The remaining 12 barrels were stored
at WES. At ERL-N, the contents of each barrel were completely homog-
enized, wet sieved prior to use through a 1-mm mesh sieve to remove
large particles, and distributed to 3.8-L brown reagent bottles.
During the distribution process, the sediment was repetitively mixed.
To ensure that the contents in the bottles were consistent, 400-ml
samples were taken from before the 1st, 25th, and 50th bottle for
moisture content and chemical analysis. Each bottle was coded with
barrel number, date, and the name of the person to whom the material
was assigned, and then stored at 4°C.
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MAINTENANCE
DREDGING
Figure 3. Black Rock Harbor, Connecticut,
source of dredged material
Sediment Dosing System
11. Two identical sediment dosing systems were constructed to
simultaneously provide either BRH or REF as suspended sediment to several
toxicity tests (Figure 4). The dosing systems consisted of conical-shaped
slurry reservoirs placed in a chilled fiberglass chamber, a diaphragm
pump, a 4-L separatory funnel, and several return loops that directed the
particulate slurry through dosing valves. The slurry reservoirs (40 cm
diam by 55 cm high) contained 40 L of slurry comprised of 37.7 L of
filtered seawater and 2.3 L of either BRH or REF sediment. The fiberglass
chamber (94 cm x 61 cm x 79 cm high) was maintained between 4° and 10°C
16
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using an externally chilled water source. (The slurry was chilled to
minimize microbial degradation during the test.) Polypropylene pipes
(3.8 cm diam) placed at the bottom of the reservoir cones were
connected to Teflon® diaphragm pumps (16 to 40 L/min capacity).
This type of pump was used to circulate the slurry but minimize
abrasion so that the physical properties and particle sizes of the
material remained as unchanged as possible. The separatory funnel was
connected to the pump and returned to the reservoir by polypropylene
pipes. The separatory funnel served two functions: (a) to ensure that
a constant head pressure was provided by the overflow, and (b) to
serve as a connection for the manifold located 4 cm below the constant
head level. The manifold served to distribute the slurry by directing
a portion of the flow from the funnel (through 6 mm inside diameter
polypropylene tubes) through the Teflon® dosing valves (Figures 4 and-
5) and back to the reservoir. At the dosing valves, the slurry was
mixed with seawater for the mussel contaminant uptake study and the
acute toxicity studies. Argon gas was provided at the rate of 200
ml/min to the reservoir and separatory funnel to minimize oxidation of
the sediment/seawater slurry. Narragansett Bay seawater filtered (to
15y) through sand filters was used for the contaminant uptake and
toxicity studies. The dosing valves were controlled by a microprocessor
connected to a transmissometer (Figure 5). The microprocessor was
programmed to deliver a pulse with a duration of 0.1 sec up to continu-
ous pulse delivery and at intervals from once every second to once every
hour. Under transmissometer control, the microprocessor responds by
modulating the pulse length to achieve the desired setpoint of suspended
17
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sediment measured as turbidity (Sinnett and Davis 1983). The trans-
missometer-microprocessor system was used to control the suspended
sediment concentrations in the mussel contaminant uptake study and
the acute toxicity studies.
ARGON
INJECTION
SEPARATORY
FUNNEL
DELIVERY
MANIFOLD
I
DOSING
VALVE
TO EXPOSURE
SYSTEM
RETURN
MANIFOLD
SLURRY
RESERVOIR
CHILLED
WATER BATH
Figure 4. Sediment dosing system with chilled water
and argon gas supply
18
-------
STRIP CHART
RECORDER
MICRO-
PROCESSOR
CONTROL
BOX
I
SLURRY
DOSING VALVE
RETURN TO
RESERVOIR
SOLENOID
EXPOSURE SYSTEM
\
F±gure
TRANSM1SSOMETER
Suspended sediment feedback control loop
and strip chart recorder
19
-------
PART III: CHEMICAL CHARACTERIZATION METHODS AND MATERIALS
Contaminant Uptake Test
Mussel collection
12. The blue mussel (Mytilus edulis) was used to determine the
bloavailability of certain contaminants within BRH materials In a 28-day
test. One month prior to exposure, mussels were collected from a well
studied area in Narragansett Bay, R.I., that was relatively free of con-
taminants (Phelps et al. 1983; Phelps and Galloway 1980). Test organisms,
50 to 70 mm shell length, were temperature acclimated from 5° to 15°C at
the rate of 1°C per day, then held in unfiltered, flowing seawater until
initiation of the experiment.
Exposure system
13. The system used to expose blue mussels to BRH material in
the 28-day flowing seawater test is shown in Figure 6. The exposure
apparatus consisted of a fiberglass resin-coated plywood tank (123-L
capacity) partitioned into two compartments. Filtered seawater entering
the mixing chamber at 2 L/min was vigorously combined with the BRH
material and the mussels' food, marine algae (a mixture of Phaeodactylum
tricornutum and T-Isochrysis galbana). The mixture cascaded over a
partition into the exposure chamber containing the mussels and a trans-
mi ssometer which measured the amount of suspended particulates in the
water. To ensure that the particles were rapidly and evenly dispersed
throughout the tank, water was collected through a manifold near the
20
-------
transmissometer ana returned to the mixing chamber at a rate of 38
L/min. Polypropylene or polyethylene plumbing materials were used
throughout.
SEAWATER/SEDIMENT
SLURRY
ALGAE
TO MICROPROCESSOR
TRANSMISSOMETER
MIXING
CHAMBER
RECIRCULATING
PUMP
Figure 6. Blue mussel (Mytilus edulis) contaminant uptake system
14. The sediment dosing system delivered BRH sediment directly
into the mussel exposure chamber via the dosing valve which was con-
trolled by the microprocessor and transmissometer. As the mussels
removed the suspended particles below the desired concentration, the
microprocessor opened the dosing valve to deliver the BRH suspension
and simultaneously turned on a peristaltic pump to deliver algae to
21
-------
the chamber. Delivery volumes by the valve and peristaltic pump were
adjusted to maintain a constant ratio of sediment and algae during a
microprocessor pulse. In response to a transmissometer signal every 5
min, the microprocessor modulated the pulse length to achieve an exposure
concentration in the chamber of 9.5 mg/L of suspended particles, con-
sisting of 9 mg/L sediment and 0.5 mg/L algae (30 x 106 cells/L). This
concentration of suspended sediments was estimated to be below the
concentration that would stress or adversely affect the organisms.
A preliminary test demonstrated no appreciable mortality, histopatho-
logical responses, or adverse changes in scope for growth (SFG) after
2 weeks of exposure to 20 mg/L.
15. The control for this experiment was designed to verify that
contaminants observed in the mussels were accumulated from BRH material
rather than from the seawater or the algal cultures. The control expo-
sure was conducted in an identical test apparatus, except that no sedi-
ment suspension was delivered to the chamber. Instead, a suspended
particulate concentration of 0.5 mg/L consisting entirely of algae was
maintained by the microprocessor feedback system.
Experimental conditions
16. Test methods. Whenever possible, the general bioconcentration
test methods used were from "Proposed Standard Practice for Conducting
Bioconcentration Tests with Fishes and Saltwater Bivalve Molluscs,"
(American Society For Testing and Materials (ASTM) 1980a). Although not
specifically intended for suspended sediment testing, the general recom-
mendations defining test animal care, handling and acclimation procedures,
22
-------
seawater quality, and acceptable exposure conditions were suitable for
this test.
17. Prior to placing animals in the test chambers, 20 mussels
were randomly selected from the mussel holding system for organic and
inorganic chemical analysis to determine the baseline residues in the
mussels before the exposures began. At the start of the 28-day uptake
study, 300 mussels were placed in each of the BRH and control chambers.
During the test, 20 mussels from the BRH chamber were sampled for chem-
ical analysis on days 7, 14, and 28. Twenty mussels from the control
chamber were sampled on day 28.
18. From each sample, eight mussels were frozen (-20°C) whole for
possible future use. The remaining twelve mussels were separated into
three groups of four each, the soft tissues removed and homogenized. A
2-g sample from each of the homogenates was used for inorganic analyses
and the remainder for organic analyses.
19. Suspended particulate concentrations. Twice each week sus-
pended particulate concentrations from the control and exposure chambers
were analyzed by dry weight determination and by electronic particle
counting (ly to 40p particle range). The dry weight determinations
were conducted according to Standard Methods (American Public Health
Association (APHA) 1976) with the following modifications. The filters
were washed with a 50-ml aliquot of delonized water before sample filtra-
tion, and followed by three 10-ml rinses of deionized water immediately
after sample filtration to remove salt. Measurements of dissolved
oxygen salinity, temperature, and ammonia-nitrogen were made to docu-
ment water quality (Table 1). All of the water quality measurements
23
-------
were well within the guidelines established by ASTM. Mortality
over the 28-day period was 13% for the exposed mussels and 6% for the
controls. The dry weight of suspended particulates in the exposure
tank did not fluctuate from the nominal level of 9.5 mg/L by more than
15%. Dry weight of suspended particulates in the control tank exceeded
the nominal level of 0.5 mg/L by an average of 1.2 mg/L; however, the
total number of particles (26 x 106 particles/L) within the size
range of the algal species used and of the size which mussels filter
efficiently was within 2% of the nominal level of 30 x 106 particles/L.
This discrepency appeared to be due to mussel fecal pellets suspended
in the water samples taken for dry weight measurements. Maintenance
of the exposure system required routine cleaning of tanks and replace-
ment of BRH sediment in the sediment dosing system. On these occasions,
the system had to be shut down and the concentration of suspended
particulates did not remain within ASTM guidelines; however, this
condition comprised only 1% of the 28-day exposure period.
24
-------
Table 1
Summary of Experimental Conditions for the Contaminant Uptake
Study with Mytilus edulis*
Parameter
Control
Suspended solids,
dry wt, mg/L
Particle density,
No./L
Temperature, °C
Dissolved oxygen,
mg/L
Salinity, ppt
Un-ionized ammonia,
1.72 ± 0.18
(1.45 - 2.02)
2.6 + 0.3 X 107
(2.0 - 3.1 X 107)
15.7 ± 0.4
(15.0 - 16.4)
7.5 ± 0.6
(7.0 - 8.5)
28.4 ± 1.8
(24 - 30)
2.9 ± 1.29
(0.64 - 5.40)
Exposure
9.32 ± 0.58
(8.19 - 10.33)
12 ± 1.3 X 107
(9.6 - 13.7 X 107)
15.6 ± 0.3
(15.4 - 16.4)
7.6 ± 0.4
(7.1 - 8.4)
28.4 ± 1.8
(24 - 30)
3.83 ± 1.68
(1.04 - 6.40)
* Tabular values are mean and standard deviation (N = 9) with the range
denoted by parentheses.
Sample Preparation
Mussels
20. Organic. The analytical procedures described below repre-
sent the state-of-the-art in marine organic analysis and have been
intercalibrated with several oceanographic laboratories. EPA recog-
nized analytical methods, while available for these classes of contam-
inants, have been developed primarily for freshwater and wastewater
systems. These methods required extensive modification and intercali-
bration when applied to marine systems for the types of matrices and
levels of detection required in this study.
25
-------
21. Each of: the separate sample homogenates from above was
treated as a separate sample with appropriate blanks carried through
the entire procedure. To each sample was added 15 ml of acetone and
the mixture homogenized for 20 sec and then centrifuged at 1750 rpm
for 5 rain. The fluid layer was decanted into a 1-L separatory funnel
containing 150 ml of pre-extracted water. The acetone extraction and
centritugation were repeated once more and the extracts combined in
the separatory funnel. The extraction and centrifugation were repeated
twice more using 25 ml of Freon® 113 as the solvent. Because of the
density of Freon®, the solvent was withdrawn from the bottom of the
centrifuge tubes using a syringe. The Freon® extracts were combined
in the separatory funnel which was then shaken and the Freon® layer
drawn off. The remaining aqueous layer was extracted twice more with
50 ml of Freon® each time. The Freon® extracts were combined and the
aqueous layer discarded.
22. To remove interfering biogenic material and some residual
particulates, the combined Freon® extracts were passed through the first
column (2 x 25 cm of 100% activated 100 to 200 mesh silicic acid). For
sediment samples 2.5 cm of activated copper powder was added to the top
of the first column to remove elemental sulfur. The column was then
rinsed with 25 ml Freon® followed by 50 ml of methylene chloride. The
eluate was collected and volume reduced in a round bottom flask fitted
with a Kuderna-Danish and 3-ball Snyder column. The solvent was ex-
changed to hexane as the sample approached 5 ml. Final volume reduction
to 5 ml was accomplished by placing the sample in a concentrator tube
26
-------
and having it blown down with a gentle stream of helium (ultra-high
purity).
23. The 5-ml sample extracts were then charged onto a 0.9 cm x
45 cm second column of 5% deactivated 100 to 200 mesh silica gel.
Three fractions were collected from the column. Fraction 1 (PF-50)
consisted of 50 ml of pentane; fraction 2 (F-2) consisted of 35 ml of
20% methylene chloride in pentane; and fraction 3 (F-3) consisted of
35 ml of methylene chloride. The PF-50 fraction was designed to col-
lect the PCB's and related materials, while fraction F-2 was designed
to collect aromatic hydrocarbons. The F-3 fraction collected more
polar material, which will be analyzed in detail at a later date.
Each column fraction was reduced in volume by Kuderna-Danish evapora-
tion as above, with the solvent changed to hexane. The final sample
volume- of 1 ml was achieved by adding 1 ml of heptane to the sample in
a 10-ml concentrator tube. Glass ebullators and microsnyder columns
were added and the samples reduced on a tube heater at 110°C to 1 ml.
The extracts were then divided in half between sealed glass ampules
for archival storage and screw cap vials for gas chromatographic and
mass spectrometric analysis.
24. All glassware used for the collection, storage, extraction,
and analysis of samples was washed with Alconox®, rinsed 4 times with
hot tap water, 4 times with deionized water, capped with aluminum foil,
and muffled for 6 hours at 450°C. Immediately prior to use glassware
was rinsed 3 times with an appropriate solvent.
27
-------
25. Stainless steel centrifuge bottles were washed as glassware
and then rinsed twice with methanol, twice with methylene chloride, and
twice with hexane immediately prior to use.
26. Polytrons® used for homogenization and extraction samples
were washed as glassware and then placed in an ultrasonic bath in
graduated cylinders filled first with methanol, methylene chloride, and
then with hexane prior to use.
27. Glass fiber filters were placed individually in aluminum
foil and muffled for 6 hours at 450°C. The stainless steel filter
housing was washed and rinsed with acetone and hexane prior to use.
28. Inorganic. From each sample homogenate, described in para-
graphs 20-27, about 2 g of wet tissue was taken for inorganic analysis,
placed in a tared beaker, and weighed. The samples were oven dried at
110°C for 2 days, cooled in a desiccator, and weighed. Ten milliliters
of reagent grade nitric acid was added to each sample, which was then
allowed to digest at room temperature in a hood for 24 hours. The
samples were heated at 60°C for several days until complete dissolution
of the sample had occurred. The samples were then evaporated to near
dryness at 90-95°C, and cooled to room temperature. Three milliliters
of 30% hydrogen peroxide was slowly added in 1-ml increments to reduce
the intensity of the effervescent reaction. The solutions were then
heated to 60°C for 24 hours, evaporated to near dryness, and cooled to
room temperature. At this point the clear and colorless solutions
were transferred to 25-ml volumetric flasks with several rinses of 5%
nitric acid, and were diluted to the mark with 5% nitric acid. The
solutions were then transferred to screw cap polyethylene bottles.
28
-------
Fifty-milliliter quantities of trace metal stripped seawater (Davey
et al. 1979) were treated like mussel samples in order to estimate the
metal blank values for the Polytron® homogenization procedure for the
mussel samples. All procedures used to prepare the glass centrifuge
tubes and Polytron® were identical to that used for mussel samples.
The seawater remained in contact with the operating Polytron® for the
same period of time required for homogenization of the mussel samples.
Three 10-ml quantities from each centrifuge tube were then pipetted
into beakers and processed like the 2-g (wet weight) homogenized mussel
tissue.
Sediments
29. Organic. The methods which follow were used for the extrac-
tion and analysis of BRU sediment. Approximately 10 g of wet sediment
was placed in a stainless steel centrifuge tube, and 50 ml of acetone
was added. The mixture was homogenized for 40 seconds using a brass-
bearing equipped Polytron®, and then centrifuged at 10,000 rpm for 5
minutes. The acetone was decanted in a 1-L separatory funnel containing
150 ml of pre-extracted deionized water. The extraction and centrifu-
gation steps were repeated once more and all extracts were combined in
the separatory funnel. The aqueous layer in the separatory funnel was
extracted three times with 50 ml of Freon® 113 each time and the extracts
were combined in a 500-ml Erlenmeyer flask. Extracts were frozen to
remove water.
30. The sediment Freon® extracts were subjected to the same two
column chromatographic separations as were the tissue sample extracts,
29
-------
except tor the addition of a 2.3-cra activated copper powder layer to
the first column for the removal of elemental sulfur. The copper
powder was activated by washing it with feN KCl, followed by a deionized
water rinse and then a raethanol and raethylene chloride rinse. The
first column removed biogenic material and the second column separated
the sample into the non-polar PF-50 fraction which contained PCB's and
other similar materials; an F-2 fraction, which contained primarily
aromatic hydrocarbons; and an F-3 fraction, which contained more polar
material and will be analyzed later. The samples were reduced in volume
and split for analysis and archival storage as described above. Similar
analytical methods have been reported by Lake et al. (1979).
31. Inorganic. After thoroughly homogenizing the sediment
contained in a barrel (see homogenization of sediment) nine samples
were taken for analysis from each barrel. These nine samples included
three from the top, three from the middle, and three from the bottom.
The wet weight of each sample was determined. Samples from barrel #00
were ladled into 400-ml Pyrex beakers and samples from barrel #LL were
ladled into 250-ml acid-cleaned polyethylene bottles. The wet weight
of all samples was then determined. The samples were frozen and then
freeze dried in a Virtis® lypholyzer (model #10-145MR-BA) for 2 days.
The dry weight of each sample was then determined.
32. Samples from barrel #00 were acidified with a total of 50 ml
of concentrated HN03 (reagent grade). The acid was added in 10-ml
aliquots since BRh sediment is very reactive to acid. All reaction was
allowed to subside before the next addition of acid was made. After
several days the samples were heated at 60°C. The sediment samples
30
-------
were subsequently evaporated down to approximately 10 ml after which
30% H202 was added in 2-ml aliquots until 50 ml had been added. The
H202 was added cautiously since BRH sediment reacts vigorously with
strong oxidizing agents. The samples were evaporated down to approxi-
mately 25 ml and filtered through acid-rinsed (5% HN03> Whatman® 41
filter paper into 250-ml volumetric flasks. The beakers were rinsed
with 25-ml quantities of 5% HM^. The rinse solution was also filtered
through the filter paper and added to the volumetric flask. The volumet-
ric flasks were brought up to volume with 5% HN(>3. The solutions
were then transferred to polyethylene bottles fitted with polyethylene
screw caps. Two empty beakers were taken through the entire concen-
trated acid dissolution procedure to estimate applicable metal blanks.
33. Dilute NH03 (5%) was added to the 250-ml bottles containing
sediment from barrel #LL and allowed to stand at room temperature for
1 week. The caps were loosely placed on top of each bottle during the
first few hours since gas (probably H2S) is liberated during this
elution process. The bottles were shaken vigorously once each day.
After 1 week the samples were filtered through acid-rinsed Whatman® 41
filter paper into acid-cleaned polyethlene bottles. No H202 was added
to these samples. Two empty bottles were taken through the entire
dilute acid dissolution procedure to estimate applicable metal blanks.
The two dissolution techniques were used to determine if different
metal concentrations would be obtained for BRH sediment samples.
31
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Sample Analysis
Organic
34. Electron capture gas chromatographic analyses were
conducted on a Hewlett-Packard model 5840 gas chromatograph equipped
with a 30-m DB-5 fused silica capillary column from J&W. The chromatograph
was temperature programmed from 80°C to 290°C at 10°C/min with a 4-min
hold at 80°C. Flame ionization gas chromatographic analyses were
conducted on a Carlo Erba Fractovap gas chromatograph also equipped
with a 30-m DB-5 fused silica capillary column from J&W. The temperature
was programmed from 60°C to 325°C at 10°C/min with a 4-min hold.
35. Gas chromatograph/mass spectrometric (GC/MS) analysis was
conducted on a Finnigan model 4500 equipped with a J&W DB-5 30-m fused
silica capillary column. The capillary column was connected directly
to the mass spectrometer with no interface present so that the effluent
from the column passed directly into the ionization source of the mass
spectrometer. The mass spectrometer was operated through a standard
Incos data system and was tuned to meet EPA quality assurance specifi-
cations using decafluorotriphenylphosphine. The ionizing current was
typically set at 300 uA and 70 V, and the instrument operated such
that 100 pg of polynuclear aromatic hydrocarbons from naphthalene to
benzopyrene gave easily quantifiable signals on their molecular ions
with signal-to-noise ratios of 50:1 or better. The mass spectrometer's
gas chromatograph was programmed from 50°C to 330°C typically at
10°C/min with a 2-min hold at 50°C, but was occasionally programmed at
4°C/min for higher chromatographic resolution.
32
-------
36. All instruments were calibrated with authentic standards
each day quantitation was attempted. The concentrations of the stand-
ards used were chosen to be close to the levels of the materials of
interest, and periodic linearity checks were made to ensure the proper
performance of each system. In some cases, authentic standards were
not available, such as for the alkyl homologs of the aromatic hydrocar-
bons. In this case, the numbers reported are a low estimate of the
actual amount present, because the response factors for these homologs
were assumed to be equal to those of the corresponding PAH's, and the
alkylated homologs all have a decreased molecular ion intensity compared
to the corresponding PAH's.
37. We also did not have standards for biphenyl, acenaphthene,
fluorene, and the aromatics with molecular weights greater than 252.
Since the molecular weights of biphenyl (154), acenaphthene (154), and
fluorene (166) lie between those of naphthalene (128) and anthracene
and phenanthrene (MW's 178), the response factors of these three com-
pounds were estimated by averaging the response factor of naphthalene
with the average of the response factors of phenanthrene and anthracene.
Response factors for the aromatics heavier than the 252's were taken to
be the same as that for the 252's.
Inorganic
38. All flame atomization (FA) atomic absorption (AA) analysis
was done with a Perkin-Elmer atomic absorption instrument (Model 603).
All Hg determinations were done by the method of Hatch and Ott (1968)
using a Perkin-Elmer mercury/hydride system (Model MHS-1) adapted to
33
-------
the 603 AA. The transient Kg signals were recorded with a Perkin-Elmer
strip chart recorder (Model 56). All heated graphite atomization
(HGA) atomic absorption determinations were conducted with a Perkin-
Elmer HGA unit (Model 500) coupled to a Perkin-Elmer atomic absorption
instrument (Model 5000) retrofitted with a Zeeman HGA background correc-
tion unit. The Model 500 HGA unit was equipped with an auto injector
(Model AS-40). The transient HGA-AA signals were recorded with a
Perkin-Elmer strip chart recorder (Model 56) and also sent automatically
to a Perkin-Elmer data station microcomputer (Model 3600). Software
supplied with the data station reduced the transient signals to a peak
height and peak area for each element determined. The instrument
set-up procedures for the FA-AA, MHS-1, and HGA-AA determinations
were in accordance with procedures described in "Methods For Chemical
Analysis of Water and Wastes" (EPA 1979) and are also found in the
manufacturer's reference manuals.
39. The AA instruments were calibrated each time samples were
analyzed for a given element. Instrument calibrations were generally
checked after every five samples had been atomized into the flame unit,
injected into the HGA unit, or pipetted into the MHS-1 sample reaction
flask. All samples were analyzed at least twice to determine signal
reproducibility; most were analyzed three times. Generally one sample
was determined by the method of standard addition, and one procedural
>
blank sample was analyzed for each 15 samples processed.
40. All elements (i.e., Fe, Zn, Mn, Cu, Pb, Cd, Cr, and Ni)
except Hg and As were determined in the sediment samples by FA-AA.
Mercury was determined only in the BRH barrel #00 samples by the
34
-------
MHS-l-AA technique. Arsenic could not be determined in the sediment
samples because of a chemical interference. At this time the cause of
the chemical interference is under investigation.
41. Due to the limited sample size of the mussel samples (i.e.,
2 g wet weight), only Fe and Zn could be determined by conventional
FA-AA. All other elements (i.e., Mn, Cu, Pb, Cd, Cr, and As) were
determined by HGA-AA. All mussel samples determined by HGA-AA were
matrix matched before analysis. A matrix solution containing 10%
seawater and 90% 0.16 N nitric acid (V/V) was used as a diluent for
both standards and samples. Samples were diluted with this matrix
modification solution so that the sample extracts never exceeded 20%
of the total volume of the solution analyzed. Standards were made up
in an identical manner to the samples.
35
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PART IV: BIOLOGICAL CHARACTERIZATION METHODS AND MATERIALS
Overview
Methods
42. The 4-day (96-hr) and 10-day flow-through toxicity tests
described below generally followed the methods prescribed by "Standard
Practice for Conducting Acute Toxicity Tests with Fishes, Macroinverte-
brates, and Amphibians" (ASTM 1980b). Although these acute toxicity
test methods were not specifically designed for suspended sediment or
solid phase sediment tests, they provided recommendations for test
animal care, handling, and acclimation, as well as guidelines for
experimental designs, water quality parameters, statistical analyses,
and general quality criteria that were suitable for the sediment tests.
Holding and acclimation conditions for each species were similar or
identical to the test conditions (Table 2).
43. Toxicity tests. For each species, two types of flow-through
toxicity tests were conducted:
£. Solid phase, in which BRH sediment or REF sediment was
placed in the bottom of the exposure chambers and fil-
tered seawater allowed to flow over the sediment
b_. Suspended particulate phase, in which suspensions of the
sediments (25 rag/L) were dosed in combination with a
solid phase sediment (either REF or a no-effect percent-
age of BRH as determined by the solid phase tests).
The solid phase tests represent conditions similar to that on the dis-
posal mound in an undisturbed (quiescent) state; the tests combining
solid phase and suspended sediments represent the conditions on the
disposal mound in a dynamic state.
36
-------
Table 2
Test Species, Exposure Conditions, and Food Used for Solid Phase
Species
Annelids
Neanthes
ar cnaceode n t ata
Nephtys incisa
Molluscs
Yoldia
llmatula
Mulinla
lateralis
Arthropods
Mysidopsia
bahia
Anpelisca abdita
Fishes
Menidia
menidia
Cyprinodon
variegatua
Ammodytes
americanus embryo
larvae
Paralichthys
dentatus
Pseudopleuronectes
americanus
N*
3
3
2
2
3
6
2
2
2
6
2
2
Biological
Exposure Time
days
10
10
10
10
4
4
4
4
4
4
4
4
Assays for
Salinity
ppt
30
(30-32)**
31
(30-32)
28
(27-30)
29
(18-30)
29
(26-30)
30
30
(30-30)
29
(28-30)
30
(30-30)
29
(26-32)
28
(26-30)
29
(26-32)
Sediment Characterization
Temperature
°C
19
(19.8-20.1)
20
(19-21)
19
(18-21)
19.5
(18-21)
25
(23-26)
20
(19-21.5)
20.5
(20-20.8)
21.2
(20.2-21.9)
10
(9-11.5)
10.5
(8.2-11.2)
20
(19.5-20.8)
10
(9-11)
Se water Flow
Kate, ml/min
51
52
50
45 to 80
35
45 to 80
45
45 to 50
34
21 to 34
43 to 49
35 to 40
Photoperiod
hrs
14
14
14
14
12
14
12
12
12
12
12
12
Food
none
none
none
none
Artemia
nauplii
none
Artemia
nauplii
none
none
Brachionus
glleatlllB
Artemia
nauplii
Artemia
nauplii
* N - number of tests conducted.
** Parentheses denote ranges.
-------
44. In the solid phase tests, a measured quantity of BRH, REF,
or a mixture of the two sediments was placed in an exposure chamber
and filtered seawater allowed to flow over the material. Introduction
of the test animals into the exposure chambers was delayed 2 to 24 hr
after starting the seawater flow to allow settling of any suspended
material. After the animals were placed in the chambers, the sediments
were left undisturbed until the end of the test. Test species, exposure,
conditions, and food used in the solid phase tests are listed in Table 2.
45. All tests (solid and suspended sediment phase) were conducted
with sand-filtered Narragansett Bay seawater at approximately 30 ppt
salinity. The photoperiod simulated a natural cycle for the time of
year these tests were conducted (Tables Bl to B52). Test temperatures
were generally held at 20°C; however, two species (Ammodytes and
Pseudopleuronectes) required 10°C.
46. The performance of suspended particulate phase tests required
the interfacing of three experimental modules: the suspended sediment
(SS) controlled dosing system, the SS dilution system, and the SS
toxicity test system. The suspended sediment dosing system (Figure 4)
supplied REF and BRH sediments at a concentration of 5.75 percent
(2.3 L of sediment in 37.7 L of seawater) to the suspended sediment
dilution system (Figure 7). This system consists of individual (REF
and BRH) 72-L glass aquaria containing a transmissometer to measure
turbidity and a recirculating submersible pump and manifold to maintain
a uniform particle distribution. Temperature-controlled filtered
seawater was introduced into the aquaria on demand. A suspended sedi-
ment concentration (BRH and REF) of 25 mg/L was maintained for all
38
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acute tests by the microprocessor-transmissometer feedback loop (Figure
5) that controlled the pulse duration of the dosing valves of the sus-
pended sediment dosing systems (Figure 4). From the aquaria, uniform
25-mg/L suspensions of sediment (REF and BRH) under constant head pressure.
flowed by gravity through polypropylene tubing to distribution chambers
(Figure 7). The distribution chambers, 3.8-L glass jars, had nine ports
to distribute the 25-mg/L suspensions to eight separate exposure chambers.
The ninth port, an overflow, functioned to maintain a constant head pres-
sure. A seawater distribution chamber containing temperature-controlled,
filtered seawater without sediment was provided as an additional control
for all experiments.
SLURRY
SEAWATER
RETURN TO
RESERVOIR
DILUTION
SYSTEM
TO MICRO-
PROCESSOR
TRANSMISSOMETER
DISTRIBUTION
CHAMBER
x\ n~~\
TO EXPOSURE SYSTEMS
OVERFLOW
TO DRAIN
EXPOSURE
CHAMBERS
TO DRAIN
SEDIMENT
Figure 7. Suspended sediment dilution system, distribution chamber, and
exposure chambers used for acute toxicity tests
39
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47. Statistics. Toxicity test data were analyzed by probit anal-
ysis (Finney 1971) if a dose-response relationship could be defined in
the tests. In cases where only two treatments were tested, a Chi-square
test was used to determine whether significant differences (P<0.05) in
mortality or a sublethal effect were observed.
Annelids
Collection, culture, and holding
48. Two species of polychaete annelids, Nephtys incisa and
Neanthes arenaceodentata Moore (Nereis acuminata Ehlers), were used
for the acute tests. Nephtys incisa were collected with a Smith-
Mclntyre dredge from the South reference site (Figure 2) in August and
October 1982 and February 1983. The worms were sieved (0.5 mm) from
the sediment on board ship, sorted into size classes, placed into
sediment from that station, and transported back to ERL-N. Worms were
held in the sediment with filtered Narragansett Bay water flowing over
them. If temperature acclimation was needed, the seawater temperature
was raised about 2°C/day until the test temperature was reached and
then held for at least 10 days at that temperature prior,to testing.
49. Nephtys incisa were fed prawn flakes (ADT-Prime®, Aquatic
Diet Technology, Inc., Brooklyn, N.Y.) directly on the sediment surface
during holding. At the start of the test, worms were sieved (0.335-mm
mesh seive) out of the sediment and placed in the test chambers. All
tests with N. incisa were conducted with juveniles. Neanthes
arenaceodentata were from laboratory cultures (original stock from
40
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D. J. Reish, California State University, Long Beach, Calif.). Worms
were cultured at 20 + 1°C and fed prawn flakes. Either adults or juve-
niles were used for toxicity tests.
Solid phase tests
50. The solid phase tests were conducted in glass crystallizing
dishes (150 x 75 mm). Each dish contained a 100-ml glass beaker (48 x
67 mm) in the center of the dish. The inflow water (sand-filtered
Narragansett Bay seawater) was directed into this beaker (which con-
tained no sediment), flowed out of the beaker over the sediment surface,
and overflowed the edge of the crystallizing dish. Flow rates were
approximately 50 ml/min. Each dish contained 400 ml of sediment (2.5
to 3.5 cm deep). Neanthes arenaceodentata were also exposed in indi-
vidual chambers constructed from glass Petri dishes (50 mm diameter)
with a nylon mesh collar (8 cm high) glued into the Petri dish. The
chambers were placed in a glass box that received a constant inflow of
seawater and had a siphon at the outflow to create a fluctuating water
level in the box (Pesch and Morgan 1978). The sediment layer in the
individual chambers was 1 cm deep.
51. The exposure concentrations used in the solid phase tests
were: 100, 75, 50, 25, and 0 percent BRH (100 percent REF). The mix-
tures of the two sediments were made volumetrically, mixed thoroughly,
and then distributed to the exposure chambers. An aliquot of sediment
was taken, weighed, and dried a minimum of 24 hr, and reweighed to
establish the wet-dry ratio for each exposure concentration. The
sediment was placed in the empty crystallizing dishes and the seawater
41
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turned on. After a minimum of 2 hr, the inflow water was turned off,
some water poured off so that the water level was below the edge of the
dish and the worms were added. Approximately 1 hr later, each dish was
checked to be certain all the worms had burrowed into the sediment,
then the inflow water was turned back on. Worms were not fed during
the test. Three tests were conducted with jfl. incisa, two with similar
size worms from two different collections, and one with larger worms
(Tables Bl to B3). Three tests were conducted with IJ. arenaceodentata,
two tests with adult males in the two different exposure chambers and
one test with juveniles.
Suspended particulate phase test
52. The suspended particulate tests were conducted in test cham-
bers similar to those used in the solid phase tests. Each crystallizing
dish (150 x 75 mm) contained 400 ml sediment (2.5 to 3.5 cm deep). A
smaller glass crystallizing dish (60 x 35 mm) was placed in the center
of the larger dish. The inflow water (water with suspended particulate
matter) was directed into the smaller crystallizing dish, which con-
tained no sediment. A Teflon®-coated stir bar kept the particulate
matter in suspension as it overflowed into the larger crystallizing
dish. Neanthes arenaceodentata were exposed only in the crystallizing
dishes (not individual chambers) because the results of previous
solid phase tests indicated that the crystallizing dishes were suitable
containers for this species.
53. Exposure conditions for the solid phase portion of the sus-
pended particulate tests were 100 percent REF or 100 percent BRH.
42
-------
These two solid phase exposure conditions in combination with the two
suspended sediment exposures (25 mg/L REF or BRH) gave a total of four
exposure treatments. The test was conducted four times with N. incisa
juveniles and twice with N^. arenaceodentata juveniles. The procedure
for adding the sediment and worms was the same as in the solid phase
test except for the first two tests with IN. incisa where the inflowing
seawater was not turned off when the worms were placed in the dishes
(see Results section). Worms were not fed during these tests (Table 3),
54. During the tests, all exposure chambers were examined daily
for the appearence of any worms on the surface of the sediment. On the
last day of the test (day 10), measurements were made on the burrows
visible through the side of the dishes. The depth of the deepest bur-
row and estimated average depth of the burrows were noted. Then the
sediment was sieved (0.335-mm seive mesh) and the worms retrieved and
counted. Any missing worms were presumed dead from toxic effects.
Since a weight gain in 10-day treatments was not expected, dry weights
were taken merely to establish the size of the worms used in the test.
Molluscs
Collection, culture, and holding
55. Yoldia limatula and Mulinia lateralis, the two species of
molluscs used in the acute toxicity tests, were seived out of sediments
from the South reference site on board ship in October 1982 and January
and February 1983 (Tables B10-B13). The organisms were then returned
to the laboratory where they were sorted from shell material and placed
43
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Table 3
Test Species, Exposure Conditions, and Food for Suspended Particulate
Phase Assays for Sediment Characterization
Exposure Time
Species N* days
Annelids
Neanthes 1 10
arenaceodentata
Nephtys 2 10
incisa
Molluscs
Yoldia 1 10
limatula
Mulinia 2 10
later all s
Arthropods
Mysldopsis 2 4
bahla
Ampelisca 2 4
abdlta
Fishes
Menidia 2 4
menldia
Cyprinodon 2 4
variegatus
Aamodytes 2 4
americanus
Pseudopleuronectes 2 4
americanus
Salinity
ppt
27
(25-30)**
28
(25-30)
27.5
(25-30)
27.5
(25-30)
30
(28-31)
28
(27-29) (20.
30
(30-30)
27
(25-29)
29
(28-30)
28
(26-29)
Temperature Seawater Flow
°C Rate, ml/min
20
(19.8-20.5) 99
20
(19.2-20.5) 98
20.5
(20-21) 50
20.5
(19.8-21) 50
21
(20-21) 90
21 or 8
5-21.5)(7.5-8.5) 60
20
(19.6-20.3) 80 to 90
19.5
(19.3-20.1) 80 to 90
10
(8.2-11.6) 86 to 92
10.2
(8.2-12.7) 80 to 95
Photoperiod
hrs Food
12
12
14
14
12
14
12
12
12
12
none
none
none
none
Artemia
nauplii
none
Artemia
nauplii
none
Brachionus
plicatili
Artemia
nauplii
* N - number of tests conducted.
** Parentheses denote ranges.
-------
in containers of REF sediment. The newly collected organisms were
acclimated to 20°C at the rate of l°C/day. Mulinia lateralis were fed
the diatom Phaeodactylum tricornutum daily; Tf. limatula were fed the
REF sediment.
Solid phase tests
56. Solid phase toxicity tests were conducted with Yoldia and
Mulinia for 10 days at approximately 20°C and 30 ppt. The exposure
system consisted of three 70- by 50-mm glass crystallizing dishes
containing test sediment (65 mm deep) placed on a glass rack 3 cm
off the bottom of a 3.8-L glass jar with a siphon to drain. Filtered
seawater entered the exposure system at a flow rate of 45 to 80 ml/min,
depending on the test species, and was circulated within the system by
a Teflon®-coated stir bar operated by a water-driven stirrer. Each
exposure system was placed in a 20°C water bath of recirculating
seawater and monitored with a continuous temperature recorder. Organ-
isms were added 6 to 24 hr after the sediments were distributed to the
exposure chambers. Experimental concentrations were 100% BRH and 100%
REF for Mulinea, and 100, 66, 50, 33, 25, and 0 percent BRH for Yoldia.
The water content was determined for all sediment combinations. Each
treatment consisted of two replicates. A minimum of two replications
of each test were conducted to satisfy statistical design criteria.
Suspended particulate phase tests
57. For the suspended particulate exposures, 25-mg/L BRH and REF
slurries (previously mixed in the suspended sediment dilution aquaria,
45
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Figure 7) were each delivered to a crystalizing dish (17 cm x 9 cm)
fitted with a standpipe to maintain a constant water level (Figure 8).
Material was kept in suspension with a water-driven stirrer and stir
bar. To adjust flow rates to each exposure container, U-shaped glass
siphons were set at the desired height. The suspension was collected
by small glass funnels that drained through polypropylene tubing to
the exposure chambers. Flow rates were measured daily (Table 3).
SLURRY
SIPHON
STANDPIPE
DISTRIBU
CHAMB
I
T
EF
ON
?
1
1
TO DRAIN
EXPOSURE
CHAMBER
TO EXPOSURE
CHAMBER
SEDIMENT
SUPPORT RACK
MAGNETIC
STIRRER
Figure 8. Distribution and exposure chambers used for solid phase and
suspended particulate phase exposure of Yoldia limatula,
Mulinia lateralis, and Ampelisca abdita
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58. Sediment suspensions were monitored at least twice during
each test using the dry weight measurement as described in the contam-
inant uptake test.
Arthropods
59. An estuarine mysid, Mysidopsis bahia, and a benthic amphipod,
Ampelisca abdita, were the two species of arthropods used in both the
solid phase and the suspended particulate phase toxicity tests (Tables
B16-B28).
Collection, culture, and jiolding
60. Mysidopsis bahia were cultured in the laboratory for several
generations according to methods described in Gentile et al. (1982)
under conditions identical to those used in the toxicity tests. Cul-
tures were maintained in filtered (15 ) Narragansett Bay seawater at
28+2 ppt salinity and 25° + 2°C temperature with a photoperiod of 14
hr light. Cultures were fed daily ad libitum 24-hr posthatch Artemia
salina from the reference strain (Sorgeloos 1980). Reference brine
shrimp were used because they were high in nutritional value, had a
high hatching percentage, and were low in contaminants.
61. Ampelisca abdita were collected from Long Island Sound or
Narrow River, Rhode Island, and transported unsieved to the laboratory
in native sediment. The sediments were then sieved and the recovered
animals placed in presieved native or reference sediment for acclimation.
These animals were acclimated to the test temperature (20°C) at the rate
of l°C/day. During acclimation, Ampelisca were fed daily ad libitum
47
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with the diatom Phaeodactylum tricornutum. Animal collection dates and
location are listed in Appendix Tables B21-B28.
Solid phase tests
62. Three flow-through acute toxicity tests were completed with
M. bahia. Continuous-flow bioassays were conducted with a diluter
system modified from Mount and Brungs (1967) and an exposure system
from Sosnowski, Germond, and Gentile (1979)* The exposure system
employed a siphon-flush mechanism that produced ten turnovers (volume
additions) per day (Table 2). Sioassays were maintained at a tempera-
ture of 25° + 2°C and salinity of 28+2 ppt and illumination of 1000
lux on a 14-hr light cycle. Sediment from BRH and REF (at room temper-
ature) was stirred and shaken vigorously before mixing to obtain the
desired percentage for each treatment. The sediment was added to the
exposure cups to a depth of 2 cm and allowed to remain in the system
overnight in flowing seawater before animals were introduced. Treat-
ments for all assays were: 100, 75, 50, and 25 percent BRH with a REF
control and a seawater control (no sediment).
63. The first test employed two exposure cups per replicate, for
a total of 20 animals per treatment (Table B16). Dissolved oxygen and
salinity were not monitored during this test. Cups for this test were
glass Petri dishes (100 mm diameter) with a 250-p nylon screen forming
the sides. The cups were removed daily for monitoring. Test organisms
could be seen most easily over the sediment by shining an intense beam
of light horizontally from the side of the test cup at the sediment/
water interface.
48
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64. For the second and third tests, five 24- to 30-hr postrelease
juveniles were randomly distributed into each of three exposure cups per
two replicates for a total of 30 organisms per exposure concentration.
Each cup was fed 24-hr posthatch reference Artemia salina daily and
removed after 96 hr exposure to determine mortality. Dissolved oxygen
was measured by Winkler titration on days 1 and 4 of these tests. Cups
in these tests were glass jars (70 mm in diameter x 83 mm high) with a
32-mm-diam hole on either side covered with 250-p nylon screen, and
were removed for monitoring only at the end of the test (96 hr).
65. In the solid phase toxicity tests with Ampelisca abdita
acclimated organisms were sieved from their holding sediments, outsized
and dead organisms discarded, and the remainder sequentially distributed
into 100-ml plastic beakers of 20 organisms each. At least 40 organisms
were preserved for size determination and the remaining beakers of organ-
isms were transferred to experimental chambers and checked after 1 hr
in order to replace any organisms that had not burrowed. Two exposure
chambers (40 organisms) per treatment were placed in the exposure system.
66. The amphipod exposure system was similar to that used for the
bivalve molluscs, except that each 3.8-L jar contained two exposure cham-
bers that consisted of 0.24-L glass jars, with four 2.5-cm-diam holes
covered with 0.4-mm mesh nylon screening. These small jars were fitted
with polypropylene lids and self-starting siphons. The water flowed
through the screens, into the exposure chambers, and out through the
siphons to the drains (Figure 8).
67. Exposure chambers were checked daily and the number of indi-
viduals dead, moribund, on the sediment, and on the water surface were
49
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recorded. The number of molts and condition of the tubes constructed
were also monitored. At the conclusion of each assay, the sediment in
all containers was sieved and the animals counted. Any animals missing
were assumed to be dead. LCSO's were based on records of dead animals
only.
Suspended particulate phase tests
68. Suspended sediment toxicity tests with Mysidopsis bahia were
conducted using the same test apparatus as that used for the annelid
studies (Figure 7). Test organisms used for the studies were cultured,
held, and acclimated using the same procedures as those used in the
solid phase tests. Two distribution chambers (one for BRH and one for
REF, Figure 7) delivered mixtures of filtered seawater and sediment
slurry at a final particulate load of 25 mg/L. A third distribution
chamber delivered only filtered seawater to the control treatment.
Test chambers consisted of 2-L glass culture dishes that held three
exposure cups (glass jars with netted holes as in the solid phase
tests). The exposure cups were held above the bottom of the test
chamber by a glass grid to allow a stir bar to mix the particulate
suspension. A siphon in the culture dish provided a vertical excursion
of the test suspension for additional mixing. Flow of the test suspen-
sion into the chambers, calibrated daily, was approximately 90 ml/min.
69. Bioassays were maintained at a temperature of 21° + 1°C,
salinity was 28+2 ppt, and illumination was 1000 lux on a 12-hr light
cycle. Dissolved oxygen was measured on days 1 and 4 of the 4-day tests,
The suspended particulate concentration was measured in both treatments
50
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daily with an electronic particle counter. For a period of 8 days,
particle counts were made daily from the splitters, exposure chambers,
and exposure cups to determine the consistency of the suspended particle
density. Thereafter, counts were made daily in the exposure chambers
only.
70. Five 24- to 30-hr postrelease juveniles were randomly distri-
buted into each of three exposure cups per two replicates for a total
of 30 animals per treatment. Each cup was fed 24-hr posthatch reference
Artemia salina daily, and removed after 96 hr exposure to monitor mor-
tality.
71. The suspended particulate phase Ampelisca exposure system
was similar to that used for the bivalve molluscs (Figure 8). In
addition, in these studies, intermediate exposures of BRH sediment
between 100 percent BRH and 0 percent BRH (100% REF) were used. To
achieve varying degrees of exposure to the dredged material, but still
maintain constant particle densities, a siphon and collection tube
from both suspension systems (REF and BRH) were directed to a single
exposure chamber. For example, to get one third BRH and two thirds
REF (33 percent BRH exposure) at 60 ml/min, 20 ml/min of BRH and 40
ml/min of REF would be combined. A 66 percent exposure would be just
the opposite: 40 ml/min BRH and 20 ml/min REF. Flow rates were meas-
ured daily. The bioassay monitoring schedule was identical to that
previously described for solid phase .tests with Ampelisca.
51
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Fishes
72. Five species of fishes were exposed to solid phase BRH mate-
rial and four species to suspended particulate phase BRH material. Two
species, the Atlantic silverside (Menidia menidia) and sheepshead minnow
(Cyprinodon variegatus), have been used routinely in aquatic toxicology;
the remaining three, the American sand lance (Ammodytes americanus),
summer flounder (Paralichthys dentatus), and winter flounder
(Pseudopleuronectes americanus), have not been routinely used but
represent fish species likely to be directly exposed to the disposal
of dredged material. Menidia menidia and £. variegatus are estuarine
species that spawn their demersal, adhesive eggs in salt marsh areas
during the warm months of the year. Ammodytes americanus is an early
winter spawner that deposits its eggs in or on sand substrates.
Paralichthys dentatus ranges inshore to well offshore and spawns in
the winter and produces a pelagic egg whereas P^. americanus is a near-
shore species that spawns in the early spring and produces a demersal
egg mass.
Collection, culture, and holding
73. Menidia menidia (Atlantic silverside) eggs and larvae were
obtained from field-collected adults (Succotash salt marsh, Rhode Island)
that were induced to spawn in the laboratory. The spawning method was
that described by Middaugh and Takita (1983) with some minor changes:
current velocity was not altered to mimic tidal flow in some of the
spawning tanks and a flow-through seawater system was used rather
52
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than a recirculating system. Seawater temperature was maintained at
18° to 20°C and salinity range was 28 to 30 ppt. Eggs were released
and fertilized by adults onto acrylic fiber mats approximately 20 x
20 x 10 cm. Mats containing eggs were transferred to 5-L hatching
jars within 48 hr of fertilization and gently aerated. Newly hatched
fish were initially fed rotifers (Brachionus plicatilis_), followed by
a daily feeding of newly hatched Artemia sp. (reference strain, Sorgeloos
1980) after the first 2 days. Embryo development, hatching, and larval
development took place within a temperature range of 18° to 21°C and a
salinity range of 29 to 31 ppt.
74. Cyprinodon variegatus (sheepshead minnow) larvae were obtained
from eggs spawned in the laboratory. The methods used were similar to
those by Hansen et al. (1978). Adult sheepshead, obtained from a salt
marsh on Santa Rosa Island, Escambia County, Florida, were kept in a
160-L aquarium supplied with flowing seawater (300 ml/min). The tem-
perature in the tank was 26°C (25.5° to 26.5°C) and the salinity was
30 ppt (29 to 31 ppt). The fish were contained in a nylon basket in
the tank so that the eggs spawned fell through the bottom of the basket
and onto a collecting screen below. The eggs were incubated at 20°
to 23°C. (One set of eggs, those used in replicate 1 of the second
suspended phase test, were incubated at 30°C to accelerate hatching.)
After hatching, the larvae were fed reference (Sorgeloos 1980) brine
shrimp (Artemia sp.) daily.
75. Ammodytes americanus (American sand lance) adults were col-
lected from the Merrimac River, Massachusetts, in November 1982 and
53
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transported to ERL-N.* They were maintained at ambient seawater temper-
ature in 1.1-m-diam tanks containing approximately 5 cm of sand. From
November 1982 through January 1983 the water temperature decreased
from 15° to 8°C and the photoperiod was changed from 11 hr light to
9 hr light. At 8°C the adults were spawned artificially into basins
and the eggs were coated with diatomaceous earth. The embryos were
transferred to mesh baskets suspended in basins of seawater. The
basins were maintained in an incubator at 8.0° to 10.2°C. Photoperiod
increased from 9 to 12 hr light; salinity was maintained at 28 to 32
ppt (by the addition of deionized water), and the seawater was gently
aerated. The larvae were maintained under identical .conditions and
immediately after hatching were fed rotifers (Brachionus plicatilis).
Larvae for the suspended particulate tests were provided by the NMFS
Laboratory, Narragansett, R.I., and maintained under the conditions
described above until testing.
76. Summer flounder (Paralichthys dentatus) were obtained from
natural spawnings of laboratory-held brood stock at the NMFS-Narragan-
sett, R.I. The eggs were spawned in 18°C and 30 ppt salinity seawater
and hatched 3 days later. The larvae were reared in black plastic
containers as described by Klein-MacPhee (1981). The average rearing
temperature was 14°C (13.6° to 15.2°C), salinity averaged 30 ppt (29
to 30 ppt).
* The taxonomy of the sand lance in the literature is unclear; however,
the fish collected most closely ressembled A. americanus; per-
sonal communication, Mr. Lawrence Buckley, National Marine
Fisheries Service (NMFS), Narragansett, R.I.
54
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77. Larvae were fed laboratory-cultured rotifers (B. plicatilis)
for approximately 3 weeks when they were fed newly hatched reference
strain (Klein-MacPhee, Howe11,and Beck 1982) Artemia nauplii. They
were then transferred to 47-cm-diam fiberglass tanks supplied with
filtered, flowing seawater at 16° + 1°C and 29 to 30 ppt. After meta-
morphosis, which occurred in 90 percent of the animals at 7 weeks
posthatch, the juveniles were used only in the solid phase toxicity
tests as there were insufficient animals to conduct the suspended
sediment tests.
78. Adult winter flounder (Pseudopleuronectes americanus) were
collected in Narragansett Bay, R.I., in November and December 1982.
The fish were transported to ERL-N and maintained in 2.4-m-diam tanks
provided with flowing seawater at ambient temperature and salinity
(2.4° to 12.2°C, 27 to 30 ppt). They were allowed to ripen naturally
and the fish first spawned 7 February 1983. The eggs of a single
female were stripped manually into a plastic dishpan treated with
diatomaceous earth/Co prevent clumping (Smigielski and Arnold 1972).
Incubation techniques, collection, and rearing are described in Klein-
MacPhee, Howell, and Beck (1982). Incubation temperature and salinities
ranged from 3.8° to 5.2°C and 28 to 30 ppt, respectively, and larvae
hatched 6 to 8 days postfertilization. Larval-rearing temperatures
and salinities ranged from 8.5° to 9.6°C and 28 to 30 ppt, respectively.
Winter flounder larvae were fed in a- manner similar to summer flounder
and were switched to reference strain (Sorgeloos 1980) Artemia approxi-
mately 1 month posthatch.
55
-------
79. A second batch of eggs were obtained on 23 February 1983.
Embryos and larvae were cultured as described above; incubation tempera-
tures and salinities ranged from 4.2° to 6.1°C and 28 to 30 ppt, respec-
tively. Larval rearing temperatures and salinities varied from 7.6° to
9.3°C and 27 to 30 ppt, respectively.
Solid phase tests
80. Menidia menidia solid phase tests were conducted in 19-cm-
diam by 10-cm-
-------
water quality. Larvae were fed daily with approximately 30% body wet
weight of newly hatched reference brine shrimp.
83. Cyprinodon variegatus solid phase tests were conducted as
described for Menidia menidia above. The tests were conducted at 20°
to 22°C and 28 to 30 ppt salinity, at flow rates of 45 to 50 ml/min
(Table B33 and B34). The test larvae were fed newly hatched reference
brine shrimp on day 0, 1, 2, and 3 for the first test and only on day
zero in the second test. Dissolved oxygen measurements were taken
only during the second test.
84. Ammodytes americanus embryonic solid phase tests were con-
ducted using the same general exposure chamber design described for
annelid solid phase tests. Embryos were placed inside sediment-covered,
mesh-bottomed glass tubes (45 X 15 mm) settled into 300 ml of sediment
contained in 110-mm-diam glass finger bowls, with two embryo chambers
(five embryos per chamber) per finger bowl. Two finger bowls were
submerged in a 30-L water bath (9.0° to 11.5°C and 28 to 32 ppt salin-
ity). Siphons and a central water well ensured an oxygenated flow of
water over the embryos. Two tests were conducted using seawater (SW)
and REF controls and 25%, 50%, 75%, and 100% BRH. (Tables B37 and B38).
Samples of the sediment were taken for moisture content as described
for molluscs.
85. Ammodytes americanus larval solid phase tests were conducted
in finger bowls (85 mm diameter) surrounded by 250-y mesh, creating
a column 80 to 90 mm high. The mesh column was welded together at the
seam and glued to the finger bowl with silicone adhesive. Each cup
held 100 ml of sediment and two or three chambers were placed in a
57
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water bath to a depth 10 to 20 ram below the top of the 250-^ mesh
column. Five larvae were placed per dish. Test conditions ranged
from 9.0° to 11.0°C, 26 to 30 ppt salinity, and 12 hr light (Tables
B39 to B44). Two tests were conducted using the three concentrations:
SW, 100% REF, and 100% BRH; and four using the six concentrations
mentioned under the embryonic tests (Tables B39-B44). Larvae were fed
rotifers (Brachionus plicatilis) twice daily at a concentration of 2
to 3 rotifers/ml or greater. Dissolved oxygen measurements were
taken only during the final test.
86. Paralichthys dentatus (summer flounder) larvae were exposed
to the REF and BRH sediments in the manner described above for the
Menidia menidia solid phase tests except that the fish were fed brine
shrimp at the rate of approximately 100% of their body weight per day.
In the first test the temperature varied from 20.2° to 20.8°C. The
salinity was 30 ppt throughout the test, and the flow rates varied
from 42 to 45 ml/mln (Tables B47 and B48).
87. Psuedopleuronectes americanus (winter flounder) larvae were
exposed to REF and BRti sediments in the manner described above for
the Menidia menidia solid phase tests except that all fish dead or
missing after 24 hr were assumed to have died or been lost as a result
of handling and were replaced. The flow rate in these tests was 37
ml/min and the temperature varied from 9.0 to 11.0°C. In the first
test the salinity varied from 30 to 32 ppt, in the second it varied
from 26 to 29 ppt (Tables B49 and B50).
58
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Suspended particulate phase tests
88. Menidia menidia suspended particulate tests were conducted
in the same crystallization dishes described in the solid phase tests
above, and included 600 ml/dish of BRH or REF sediment in the appro-
priate treatment dishes. Treatments used for these tests were:
seawater controls (no sediment or particulates), BRH (25 mg/L), and
REF (25 mg/L). In these tests, the inflowing seawater or seawater and
suspended particulates were introduced to each dish by the distribution
chambers previously described (Figure 7). Each inflow discharged into
a 150-ml beaker located in the center of each dish, and a spinning
stir bar maintained the particulates in suspension in the beaker. The
lip of this beaker was higher than the water level of the crystalliza-
tion dish to protect fish larvae from the mixing vortex.
89. Two tests were performed using 12 larvae per treatment, and
each treatment was replicated once. Flow rates for these tests were 80
to 90 ml/min; temperature and salinity ranges were 19.6° to 20.3°C and
26 to 30 ppt, respectively. Suspended particulate concentration was
measured by dry weight of a 100-ml water sample and two Coulter counts
from each treatment (dish) over the course of the test. Dishes were
monitored daily for temperature, salinity, flow rate, and number of
larvae live, moribund, or dead. Dissolved oxygen concentrations were
measured midway through the exposure period. Fish larvae were fed a
daily ration of 30% body wet weight of newly hatched reference brine
shrimp (Tables B31 and B32). High daily rations were needed because of
the relatively high flow rates.
59
-------
90. Cyprinodon variegatus suspended particulate tests were con-
ducted as described for Menidia menidia above at a temperature of
19.3° to 20.1°C at a flow rate of 80 to 90 ml/min. During the first
suspended particulate test the salinity ranged from 25 to 26 ppt, and
during the second test, from 28 to 29 ppt (Tables B35 and B36).
91. Ammodytes americanus suspended particulate tests were con-
ducted as described above for Menidia menidia, except 15 larvae were
used per treatment and larvae lost or dead within 24 hr of the beginning
of the test were replaced. During the two tests the temperature ranged
from 8.2° to 11.6°C, the salinity from 28 to 30 ppt, and the flow
rates from 89 to 90 ml/min (Tables B45 and B46). The larvae were 28
to 32 days old (obtained from NMFS, Narragansett) and were fed rotifers
(Brachionus plicatilis) twice daily.
92. Pseudopleuronectes americanus suspended particulate tests
were conducted as described for Menidia menidia, except that larvae
lost or dead within 24 hr of the beginning of the test were replaced.
The temperature in the exposure chambers containing no sediment in
the suspended particulate tests varied from 8.2° to 10.2°C, the temper-
atures in the exposure chambers containing sediment were slightly
higher (9.0° to il.l°C in REF and 9.8° to 12.7°C in BRH)., The flow
rate varied from 85 to 95 ml/min in the first test, and from 80 to 90
ml/min in the second. In the first test (Table B51) the salinity
varied from 26 to 29 ppt; in the second test (Table B52) the salinity
was constant at 28 ppt.
60
-------
PART V: RESULTS AND DISCUSSION
Chemical Characterization
Organic contaminants
93. Mussel analysis. Results of analyses of the 28-day exposed
mussels by GC/MS are shown by the total ion current profiles of frac-
tions PF-50 and F-2 in Figure 9. The richness of these two traces is
in marked contrast to the traces shown in Figure 10 for the control
mussels. Within the PF-50 fraction of the exposed mussels (Figure 9)
there are large quantities of naturally occurring biogenic compounds
which also show up in Figure lOa. The large humplike structure in
Figure 9 is known as the Unresolved Complex Mixture which is frequently
seen in association with petroleum contamination, and consists primarily
of hydrocarbons of an aliphatic nature (Boehm and Quinn 1977; Stegeman
and Teal 1973). The high resolution capillary column run of this
fraction was examined for unknown organic compounds of a nonaliphatic
structure. In addition to DDE and the PCB isomers containing from two
to eight chlorine atoms, evidence was found which suggests that many
unusual compounds exist in this fraction. However, even with the
extreme resolution afforded by the 4°C/min runs on 30-m capillary
columns, the coelution of many compounds effectively hinders the deter-
mination of a complete spectrum for any given compound. Although
tentative identifications were not possible, the incomplete spectra
could still be used as recognizable characteristics of the dredged
material.
61
-------
100
tn
5
1000 2000 3000
a. FRACTION PF-50
4000
Scon Number
100
CO
u
A*A—L—J&1U
1000
2000 3000
b. FRACTION F-2
4000
Scan Number
Figure 9. Total ion current profiles oi the 28-day exposed
mussels analyzed by GC-MS with a 4°/min (50-330) temperature
programming rate
-------
100
500
1000 1500
a. FRACTION PF-50
2000 Scan Number
100-
z
IOOO
2000 3000
b. FRACTION F- 2
4000
Scan Number
Figure 10. Total ion current profiles of the 28-day control
mussels analyzed by GC-MS with a 4°/min (50-330) temperature
programming rate
63
-------
94. PCB's were analyzed by both capillary column electron cap-
ture gas chromatography for the total quantity of material present
measured as Aroclor 1254, and by GC/MS for their relative chlorine
number distributions. Both results are shown in Table 4. The chlorine
number distributions were calculated from uncorrected mass spectrometer
molecular ion area measurements and so do not reflect differences in
response factors for different PCB's.
Table 4
Single Replicate PCB Concentrations as Aroclor® 1254
in
ng/g Dry Weight Including Chlorine
Number Distributions by Mass Spectrometry
Sample
Day 0
Day 28
Control
Day 7
Exposed
Day 14
Exposed
Day 28
Exposed
BRH
Sediment
Total
PCB
ng/g
74
84
2000
1100
3000
6800
di tri
0 6
2 9
37 390
34 240
58 600
130 960
Clx PCB
tetra penta hexa hepta
22 24 20 2
22 22 20 3
740 520 290 30
420 310 89 6
960 890 420 67
2500 1500 1500 130
octa
0
0
1
0
0
3
64
-------
95. Aromatic hydrocarbons form a class of compounds that are
strongly accumulated by mussels from Black Rock Harbor sediment. This
class can be divided into two groups: the parent polynuclear aromatic
hydrocarbons, and their alkylated homologs. There has been no attempt
made to quantitate each structural isomer of the parent aromatic hydro-
carbons, or to identify the particular structural or positional isomer
of their alkyl homologs. Rather, the alkyl homologs are identified as
being associated with a particular molecular weight parent PAH which
has been substituted with alkyl chains having a total of from 1 to 4
carbon atoms. These are referred to as C-l to C-4 substituents, and
could refer to any combination of methyl-, ethyl-, prppyl-, or butyl-
groups which might add up to the correct number of alkyl substituent
carbons.
96. Table 5 lists the parent PAH's found in this study, along
with their molecular weights. The molecular weights form a convenient
short-hand notation for these compounds when referring to the parent
compounds, and, with the addition of the C-l and C-4 nomenclature
mentioned above, their homologs as well. Also because of chromato-
graphic overlap, alkyl homolog distributions (AHD's) of different
PAH's having the same molecular weight cannot be separated except in
the case of the two 154's. Thus, for consistency, all PAH's of a
given molecular weight should be treated as a single measurement when
being compared with alkyl homolog distributions.
97. Appendix Tables A1-A7 list the PAH concentrations and the
concentrations of each alkyl homolog, including the variability asso-
ciated with the three replicates measured on the mussel samples.
65
-------
Table 5
Parent Polynuclear Aromatic Hydrocarbons Found in the Exposed Mussels
and Black Rock Harbor Sediment
Compound Molecular Weight
Naphthalene 128
Biphenyl 154-Bi
Acenaphthene 154-An
Fluorene 166
Phenanthrene
Anthracene 178
Pyrene
Fluoranthene 202
Benz(a)anthracene
Chrysene 228
Tripherylene
Benzofluoranthenes
Benzopyrenes 252
Perylene
Benzoperylene
Dibenzopyrene 276
and others
Dibenzanthracenes
Benzocrysenes 278
and others
Coronene
or similar 300
Dibenzocrysenes
and others 302
66
-------
Response factors for the alkyl horaologs were assumed to be the same as
the parent PAH. This leads to low but reproducible estimates of the
homolog concentrations. Also, response factors for the 154's, 166,
and those over 276 were estimated as described above. Tables 6 and 7
are summary tables of the aromatic hydrocarbon data; the average PAH
concentration for each treatment is shown in Table 6, and the concen-
tration of the sum of each alkyl homolog distribution from C-l to C-4
is shown in Table 7.
98. The total ion current profile of the day 28 control mussels
(Figure lOb) contains five peaks that have been tentatively identified
as silicones. These identifications are based on library spectra
matches and isotope ratio calculations for silicon. Spectra and reten-
tion time of authentic standards were not yet available to verify
their identifications. However, the compounds have spectra similar to
dodecamethylcyclohexasiloxane and decamethylcyclopentasiloxane. There
were a series of five of these peaks which could be measured in most
samples. Table 8 lists the results of these measurements, expressed
as arbitrary area counts per gram of mussel (or sediment) dry weight.
This allows measurements made between samples of different weight to be
compared even though there were no standards available with which valid
quantitations could be obtained. Although the exposed mussels seem to
take up these siloxane compounds, the day 28 control samples had consid-
erably more of them and the Black Rock Harbor sediment had no more
than the analytical blank. Therefore, these contaminants seem to
originate from the dosing system and not from the sediment.
67
-------
Table 6
Mean Concentrations ± Standard Deviation of Parent PAH Compounds
found in Exposed and Control Mussels and in
Black Rock Harbor Sediment in ng/g Dry Weight
PAH Concentration, ng/g Dry Weight
Compound
128
154Bi
154AN
166
178
202
228
252
276
278
300
302
Day
0
0.8
+0.4
1.3
+0.2
0.7
+0.4
1.4
+0.4
7.2
+1.6
34
+8
8.5
+2.3
15
+5
3.3
+0.7
<0.4
<0.1
<0.3
Day
7
0.5
+0.1
1.9
+0.6
3.6
+0.8
14
+1
220
+25
2600
+460
1800
+230
1300
+230
250
+67
140
+26
11
+2.8
66
+13
Day
14
1.1
+0.3
2.9
+1.0
4.7
+1.2
8.3
+1.3
140
+21
1100
+160
840
+160
610
+100
120
+22
68
+17
2.9
+0.9
17
+5
Day
28
0.6
+0.3
3.5
+2.1
3.8
+1.8
12
+3.7
180
+35
2000
+470
1800
+360
1600
+210
280
+50
160
+20
9.3
+0.8
56
+9
Day
28 Control
0.5
+1.0
5.3
+2.0
1.2
+0.2
1.2
+0.4
6.7
+1.5
19
+2.5
3.1
+1.1
5.8
+1.7
1.6
+0.9
<0.6
<0.1
<0.1
Black Rock
Harbor
Sediment
17
54
120
370
2700
7100
9800
8600
9100
4400
130
2700
68
-------
Table 7
Mean Concentrations ± Standard Deviation of the Sum of C-l Through
C-4
Alkyl Homologs of PAH
in Black Rock Harbor
I C-l through C-4
Day Day
Compound 0 7
128
154B1
154AN
166
178
202
228
252
8.6 390
+2.9 +120
9.7 710
+2.2 +92
3.0 300
+1.0 +41
14 1100
+3.4 +130
38 5600
+6.7 +990
12 3500
+1.9 +650
5.9 1500
+1.6 +290
<6 510
+100
's Found
Sediment
in Exposed and Control
Quantitated
Alkyl Homologs, ng/g
Day
14
270
+65
450
+150
160
+15
540
+110
2600
+500
1600
+290
710
+150
220
+45
Day
28
300
+100
720
+230
280
+80
1100
+250
5000
+1400
3400
+690
1800
+290
590
+79
Mussels and
as Each Parent PAH
Dry Weight
Day
28 Control
16
+2.3
27
+3.5
7.8
+0.8
23
+4.8
39
+7.1
7.7
+2.9
1.3
+0.6
<5
Black Rock
Harbor
Sediment
3400
2900
1100
3400
13000
8300
13000
4800
99. The mass spectrometric data of the day 28 mussel F-2 frac-
tion was also examined in detail for other compounds of interest which
might be hidden by the high aromatic hydrocarbon content of the sample.
The molecular weights of the aromatic hydrocarbons form a steadily
increasing elution series as the temperature of the gas chromatograph
is increased. The mass spectral data collected above the molecular
weights of the currently eluting aromatics can be examined at high
69
-------
Table 8
Distribution of Five Silicone-like Compounds in Exposed
and Control Mussels and
Black Rock Harbor Sediment Measured
as GC/MS Area Counts/Gram Dry Weight
Response of
GC/MS
Scan
Number
1277
1619
1927
2194
2432
Day
0
7200
+3300
6200
+500
28,000
+1000
10,000
+800
5700
+5500
Day
7
17,000
+1500
39,000
+2100
79,000
+6700
41,000
+4400
9900
+1000
Silicone-like Peaks
Day
14
18,000
+2500
15,000
+3200
12,000
+1700
5400
+560
2100
+210
Day
28
23,000
+5900
48,000
+17,000
86,000
+39,000
45,000
+21,000
10,000
+4100
Per Gram Dry
Day
28 Control
28,000
+2600
550,000
+120,000
1140,000
+150,000
412,000
+73,000
53,000
+13,000
Weight
Sediment
1800
1500
480
480
330
sensitivity for the presence of anomalously heavy compounds. Many of
these compounds tend to be chlorinated organics because of the increased
weight of the chlorine substituents. Several members of the DDT series
were identified in this manner as was a compound tentatively identified
as Ethylan, another chlorinated insecticide. In addition to these
identified compounds, this technique extracted the spectra of numerous
other contaminants which have yet to be identified, many of which may
contain oxygen. Although not yet identified, these spectra and reten-
tion times can serve as identification labels or fingerprints for the
same compounds in further laboratory or field studies.
70
-------
100. Sediment analysis. The Black Rock Harbor sediment material
has been analyzed for the same contaminants as decribed for the mussels,
with the exception of the anonamously heavy F-2 components. Figure 11
shows the sediment total ion current profiles for the PF-50 and F-2
fractions. Although the PF-50 fractions show some qualitative differ-
ences compared to the day 28 exposed mussel PF-50 fraction in Figure
9, the F-2 fraction is very similar to the exposed mussel F-2 shown in
Figure 10. PCB's in the sediment were measured both by capillary
column electron capture gas chromatography and by mass spectrometry.
The gas chromatographic measurements were quantitated against Aroclor®
1254, while the mass spectrometry measurements were for chlorine number
distributions. Both measurements are reported in Table 4 along with
the mussel results.
- 101. The same polynuclear aromatic hydrocarbon measurements were
made for the sediment as for the mussels. The alkyl homolog distribu-
tions in Black Rock Harbor sediment for each of the PAH's from 128 to
302 are shown with those of the mussels in Appendix A. Tables A1-A7
contain the tabulated concentrations of each PAH and each alkyl homolog
concentration from C-l through C-4. Sediment concentrations are sum-
marized in Table 9.
102. Accumulation. The results of the sediment characterization
mussel bioaccumulation study show that both PCB's and aromatic hydro-
carbons were accumulated from Black Rock Harbor sediment in as little
as 7 days. Total PCB's, measured by capillary column gas chromatography
using electron capture and quantitating against Aroclor® 1254, show a
large increase in the exposed mussels over the levels in both the
71
-------
100 n
1000
2000 3000
a. FRACTION PF-50
4000
Scon Number
100
t
CO
UJ
1000
2000 3000
b. FRACTION F-2
4000
Scan Number
Figure 11. Total ion current profiles of the Black Rock Harbor
FVP sediment analyzed by GC-MS with a 4°/min
(50-330) temperature programming rate
72
-------
Table 9
Organic Contaminants in Black Rock Harbor Sediment, ng/g Dry Weight,
Compound
128
154 Bi
154 An
166
178
202
228
252
276
278
300
302
PCB's
One Replicate
Concentration
PAH
17
54
120
370
2700
7100
9800
8600
9100
4400
130
2700
6800
AMD
3400
2900
1100
3400
13000
8300
13000
4800
-
-
-
—
pre-exposed and the day 28 controls, as shown in Table 4. Both con-
trols remained quite constant at about 80 ng/g dry weight, while the
three exposure period samples showed uptake to between 1100 and 3000
ng/g dry weight. There is an increase in the PCB levels as the exposure
time increases, although the value at day 14 is anomalously low for
unknown reasons. By the end of the 28-day uptake period, the mussels
had accumulated PCB's to a level approaching half of that in the sedi-
ment, while at day 7 they had accumulated levels that were almost one
third that of the sediment. Both sampling periods demonstrate a sizable
uptake of PCB's and indicate the clear biological availability of this
contaminant.
73
-------
103. Table 4 also shows the distribution of PCB isomers into the
various chlorine number groups from two chlorine atoms per molecule to
eight chlorine atoms per molecule. The lower chlorine number groups
correspond to PCB compounds having relatively low log P (octanol-water
partition coefficient) values, while the higher chlorine number groups
have relatively higher log P values. The two control samples, day 0
and day 28 control, are similar in the amount and pattern of their PCB
distributions, although there are some minor differences at chlorine
numbers 4, 5, and 6. These similarities are consistent with lower
Narragansett Bay mussels being held in a control tank with filtered
lower Narragansett Bay water flowing through.
104. The three exposure times also show remarkably similar PCB
patterns. The day 28 exposed sample shows slightly elevated levels of
Cl-5 and Cl-6 PCB's, but these differences are quite small. A compar-
ison of the PCB patterns of the three exposed samples with that of the
Black Rock Harbor sediment in Table 4 reveals that there are no large
pattern changes even between the sediment and the exposed mussels.
The mussels may have accumulated slightly more of the Cl-3 PCB's than
were present in the sediment, and they might have accumulated slightly
less of the Cl-6 PCB's than were in the sediment.
105. Aromatic hydrocarbons form the second class of chemical
contaminants which are strongly accumulated by mussels exposed to Black
Rock Harbor sediment. Tables 6 and 7 summarize the data for the con-
centrations of both parent PAH and for the sum of the alkyl homolog
series from C-l to C-4. The amount of parent PAH's taken up by the
exposed organisms compared with that in the sediment and the control
74
-------
organisms is shown in Figure 12, while the amount of the summed alkyl
homologs is shown in Figure 13 for the same samples. The biological
availability of aromatic hydrocarbons in the sediment is dramatic. The
concentration of the most intense PAH's, 202 to 252, shows an increase
of a factor of about 100 over control concentrations. In contrast to
these dramatic uptakes after only a week of exposure, the levels do
not markedly increase over the subsequent 21 days. As with the PCB's,
the 14-day sample is anomalously low.
10000-
5000-
< •
Z
-.-:
' <
Sediment
Day 7
Day 28
Day 14
CONTROL
T T r i i | T r r F
128 154 154 166 178 202 228 252 276 278 300 302
Bi An
MOLECULAR WEIGHT
Figure 12. Concentration of PAH compounds in Black Rock Harbor
sediment and exposed and control mussels
75
-------
13000-
10000-
5OOO-
2
128 154 154 166 178 202 228 252
Bi An
MOLECULAR WEIGHT
Figure 13. Concentration of sum of C-l through C-4 alkyl homologs
of PAH's measured in Black Rock Harbor sediment and control and
exposed mussels
106. In contrast to the PCB patterns, which showed similar uptake
for each isoraer, the PAH patterns are quite variable. The maximum
uptake is near PAH 202, where the mussels' levels are approximately one
third of the sediment levels. However, for early PAH's from 128 to
166, mussel uptake levels are only about 5% of the sediment levels and,
for PAH's above 252, the uptake may be even smaller than 5%. If one
can assume that the PAH's are all in a form equally biologically avail-
able, which has not yet been shown, then it follows that the mussels
are selecting a particular fraction of the PAH's for uptake which is
not based on log P since the octanol-water partitioning coefficient
for PAH's continues to increase as molecular weight increases.
76
-------
107. Table 7 shows approximately the same pattern for the summed
alkyl homologs as for the PAH's. The lighter PAH homologs are present
in the mussels at levels around 10% of the levels in the sediment,
while the homologs of the most concentrated PAH's, 178 and 202, are
present in the mussels at levels nearly one third those of the sediment.
Again, the heavier PAH homolog ratios decrease to about 10% of the
sediment values. The homologs of PAH's above 252 were hardly detectable
in the mussels at all. Therefore, the mussels are selecting a similar
fraction of the alkyl homologs as they did for the PAH's. Assuming
equal availability, this again is not consistent with a strict log P
uptake model.
Inorganic contaminants
108. Mussels. Due to the limited sample size (approximately 2
g of homogenized wet weight of tissue), only Fe and Zn could be deter-
mined by conventional flame atomic absorption. All other elements
(i.e., Mn, Cu, Pb, Cd, Cr, and As) were determined by heated graphite
atomization. The amount of sample available to do the mercury analysis
by the cold vapor technique was inadequate to detect low concentrations
of mercury. Nickel could not be reliably determined on this limited
sample size even with HGA-AA. The average and the standard deviation
of the average for each set of mussel samples collected from the con-
trol and exposure chambers are given in Table 10. All of the individ-
ual elemental concentrations determined for the mussel samples are
given in Table A10. The average method blank for the mussel samples
is also given in Table A10. The average method blank was
77
-------
calculated by dividing the average absolute ug value for each element
by the average homogenized mussel dry weight. All elements (except
Cu) determined in the blanks were below the detection limits of the
analysis methodology employed in this study. The percent recovery of
metal spikes added to the mussel samples prior to analysis are given
in Table All. The percent spike recovery for all elements determined
in the mussel samples was greater than 80% and less than 110%.
Table 10
Distribution of Trace Elements in yg/g Dry Weight in Exposed and
Control
Mussels
Trace Metal Concentration,
Element
Fe
SD
Mn
SD
Zn
SD
Cu
SD
Pb
SD
Cd
SD
Cr
SD
As
SD
Day
0
199
± 26
11
± 0.7
149
± 76
25
± 23
4.7
± 0.1
2.7
± 0.3
2.3
± 0.3
8.2
± 0.6
Day
7
357
± 41
12
+ 3
286
± 58
62
± 23
11
± 1.5
4.7
± 0.7
15
± 1.6
8.6
± 1.0
Day
14
330
± 6
31
± 10
160
± 18
75
± 7
8.5
± 0.9
3.1
± 0.5
12
± 1.3
7.3
± 0.4
Ug/g Dry Weight
Day
28
500
± 191
11
± 4.9
333
± 84
55 '
± 18
14
± 4.7
7.0
± 1.9
25
± 11
8.9
± 1.1
Day
28 Control
213
± 7
13
± 7
221
± 63
17
± 5
6.5
± 2.6
2.8
± 0.5
2.0
± 0.5
7.4
± 1.0
78
-------
109. Sediments. The inorganic chemical composition for the two
barrels (#00 and #LL) of Black Rock Harbor sediment is given in Table
11. The average, standard deviation of the average, and the percent
standard deviation of the average are given. The wet to dry weight
ratio is also given for the two barrels of sediment. All of the data
used to calculate these averages are given in Tables A8 and A9 for
Barrel #00 and Barrel #LL, respectively. The metal concentrations for
the two blank samples for each barrel are also given in Tables A8 and
A9. The blank metal concentrations are given in units of yg/g to
easily show the large difference between the samples and the blanks.
The average dry weight values calculated for the two sets of barrel
samples were used to convert the absolute ug values to ug/g concen-
trations for an easy comparison to the samples. No values for As are
listed in this table since a chemical interference was detected during
the analysis (for both HGA-AA and MHS-1 hydride generation techniques)
of these sediment samples. At this time the cause of the chemical
interference for the As determinations for BRH sediment samples is
under investigation. The overall results indicate that the BRH barrel
samples are reasonably homogeneous between barrrels if reasonable
precautions are taken to remix each barrel before being sampled. What
appear to be small differences between Fe and Mn for the barrels are
probably due to the two different preparation techniques that were
used to dissolve the metals that were determined. The #00 samples
were dissolved in hot concentrated nitric acid while the #LL samples
were simply eluted with 5% nitric acid at room temperature. The simi-
larity of the results indicates that the metals reported here are
79
-------
Table 11
Average Metal Concentrations in Black Rock Harbor Barrel #00 and
Element
Fe
SD
%SD
Mn
SD
%SD
Zn
SD
%SD
Cu
SD
%SD
Pb
SD
%SD
Cd
SD
%SD
Cr
SD
%SD
Ni
SD
%SD
Hg
SD
%SD
wet/dry
SD
%SD
Barrel #LL Sediment Samples*
Barrel #00
29,600
809
2
359
37
10
1200
59
4
2380
112
4
378
16
4
23.4
0.9
3.7
1430
77
5
139
4
3
1.7
0.1
4.0
3.22
.02
1
Barrel #LL
29,600
623
2
280
5
1
1210
50
4
2540
64
2
413
18
4
24.7
0.7
2.9
1300
30
2
169
5
3
-
—
—
3.03
0.17
5.6
* All concentrations are in yg/g dry weight.
and percent standard deviation are also given.
80
-------
probably all present as sulfides or some other easily solubilized
anion. Manganese has a difference of about 30% for the two barrels of
sediment. This difference is undoubtedly related to the two preparation
techniques prior to analysis. It should be pointed out that the two
preparation techniques were used simply to ascertain the magnitude of
differences that would be obtained by the two acid extraction methods.
110. Accumulation. Statistically (Student-t test, P <0.05) there
is no difference between the mean time zero mussel control samples and
the mean 28-day mussel control samples. There is no significant differ-
ence between the mean Mn, Zn, and As in the control and exposed samples
for the 28th day sampling period (P<0.05). However, there is a signif-
icant difference between the means for the 28-day control mussel samples
and the 28-day exposed mussel samples for all of the other elements
determined. The elemental ratios (calculated from the data in Table
All) of the 28-day exposure mussel samples and the 28-day control
mussel samples show that a significant uptake of several of the trace
elements had occurred in the mussel test organism. These data are
presented in Table 12. The elemental ratios of the 28-day control
mussels and the day 0 control mussel samples are also given in this
table. Any elemental ratio that approximates 1 (talcing the standard
deviation into account) indicates no significant uptake or change in
content for that element in the test organism. Essentially, all the
control sample ratios are not different from 1, if the standard devia-
tion is taken into account. For the day 28 dosed samples, however,
only Mn and As have a ratio of approximately 1. The most obvious
uptake of any of the metal ratios listed in Table 12 is Cr. The ratio
81
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of uptake (exposed/control) for this element is approximately 13.
There is no indication that equilibrium has been obtained for Cr during
this bioaccuraulation period of 28 days. The control samples, however,
show no change for Cr content during the same time period. All other
metals determined in the mussel samples tend to indicate minor amounts
of uptake.
Table 12
Ratios of Trace Metal Accumulations in Exposed and Control Mussels
28-day control ± SD
Element
Fe
Mn
Zn
Cu
Pb
Cd
Cr
As
0-day control
1.1 ±
1.2 ±
1.5 ±
0.7 ±
1.4 ±
1.0 ±
0.9 ±
0.9 ±
0.1
0.7
0.9
0.6
0.6
0.2
0.2
0.1
28 day exposed ± SD
28 day control
2.4 ±
0.9 ±
1.5 ±
3.2 ±
2.1 ±
2.5 ±
12.8 ±
1.2 ±
0.9
0.6
0.6
1.4
1.1
0.8
6.4
0.2
111. To determine if any relationships exist between any of the
metals determined for the homogenized mussel samples, linear least
square regression equations were calculated for all metals versus the
Fe concentration for each sample. In the regression line calculations,
Fe was used as the dependent variable and the other metals were made
82
-------
the independent variable. The plots are shown in Figures 14 through
20. The number of sample pairs (i.e., mussel concentrations of x and
y) must be considered in order to evaluate the significance of the
calculated correlation coefficient. In addition, the probability (P),
which indicates the possibility of a correlation coefficient being
significant due to random sampling from an uncorrelated population,
should also be considered. Values of P of 0.10 and 0.01 indicate the
possibility that 10% and 1%, respectively, of the time, a significant
correlation of x to y may occur due to random errors. All the correla-
tion coefficients used in the following discussion are based on P
values of 0.01 (i.e., 99% probability that a real correlation exists
between x and y). For example, 15 data pairs require a correlation
coefficient greater that 0.641 to be at the 1% level of significance
(Fisher 1958). The x intercept of the regression line indicates how
much of the mean Fe mussel concentration of the dependent variable (x)
is not associated with the mean independent variable (y). The slope
of the line is in effect the relative mussel concentration of Fe asso-
ciated with the total of the other metals determined. All of the data
for all the samples are plotted in these figures. The open circles
represent samples collected from the BRH exposure chamber and the solid
circles represent mussel samples collected from the control chamber.
112. At the 1% level of significance Cr, Pb, and Cd have a good
correlation with Fe. The correlation coefficients for Fe versus Cr,
Pb, and Cd are 0.978, 0.940, and 0.860, respectively. The linear least
squares regression plots for Fe versus Cr, Pb, and Cd are given in
Figures 14, 15, and 16. Qualitatively these three figures show the
83
-------
relative increase of Cr, Pb, Cd, and Fe in BRH sediment-exposed mussels
compared to these same metals in control mussel samples. The equations
for these regression lines are given below:
[Fe] = 167 + 13.5 [Cr]
[Fe] - 32.5 + 32.3 [Pb]
[Fe] + 67.3 + 62.5 [Cd]
The predictive utility of the above regression equations are not certain
at this time. However, it would be interesting to apply these equations
to metal concentrations determined in mussel samples collected from
Long Island Sound which are exposed to BRH sediment.
30
CL
£ 20
10
• Control
O BRH Exposed
200 400 600
Fe (ppm)
Figure 14. Distribution of Cr versus Fe in mussels
from the Black Rock exposure
84
-------
16
Q- 12
Q.
.o
CL
8
• Control
-O BRH Exposed
200
400
Fe (ppm)
600
Figure 15. Distribution of Pb versus Fe in mussels
from the Black Rock exposure
E
a.
a.
8
6
4
2
Control
Exposed
200 400
Fe (ppm)
600
Figure 16. Distribution of Cd versus Fe in mussels
from the Black Rock Harbor exposure
85
-------
113. The correlation coefficients for Zn and Cu with Fe are
close to the correlation coefficient required for a P value of 0.01.
The correlation coefficients for Fe versus Zn and Cu are 0.710 and
0.628, respectively. The regression plots for these elements are shown
in Figures 17 and 18. From Figure 17 it can be seen that there is
almost no increase for the Zn concentration determined in mussels
collected from the exposure chamber versus the control chamber. The
linear least squares regression equations for these plots are given
below:
[Fe] - 76.7 + 1.06 [Zn]
[Fe] = 171 + 3.16 [Cu]
IM
300
200
100
• Control
O BRH Exposed
o
200 400
Fe (ppm)
600
Figure 17. Distribution of Zn versus Fe in mussels
from the Black Rock Harbor exposure
86
-------
90
o
30
• Control
OBRH Exposed
cP
200
400
Fe (ppm)
600
Figure 18. Distribution of Cu versus Fe in mussels
from the Black Rock Harbor exposure
114. Both As and Mn have correlation coefficients that are below
the required value for a correlation at a P value of 0.01. The correla-
tion coefficients for As and Mn versus Fe are 0.589 and 0.132, respec-
tively. The regression plots for these elements are shown in Figures
19 and 20. The plots of Mn and As versus Fe demonstrate that these
two elements are of little value for showing BRH sediment uptake in
mussel test organisms. The linear least squares regression equations
for these plots are given below:
[FeJ = -349 + 82.8 [As]
[Fe] = 290 + 1.89 [Mn]
87
-------
10
E
Q.
CL
in
• Control
O BRH Exposed
200
400
Fe (ppm)
600
Figure 19. Distribution of As versus Fe in mussels
from the Black Rock. Harbor exposure
30
'e
1 20
c
2
10
Of
• Control ol
0 BRH Exposed I
/
* o
/o o
I o
1 1
200 400 600
Fe (ppm)
Figure 20. Distribution of Mn versus Fe in mussels
from the Black. Rock Harbor exposure
88
-------
115. Several of the elements showed an Increase in concentration
in the first 7 days of the uptake period. Like the uptake of the
organic compounds determined in this study, the day 14 samples appeared
to resemble a period of depuration for several of the elements measured.
A notable exception was Mn, which showed its highest concentration
during the day 14 sampling period. No explanation is easily formulated
for either this observed depuration for most of the metals determined
or the erratic uptake behaviour of Mn during this sampling date.
116. The fact that several of the elements have shown that an
uptake plateau was not reached during the study indicates that a longer
time period may be warranted to study the bioaccumulation of metals in
this marine organism. A longer time period can have two beneficial
factors. First, the amount of an element may increase in the test
organism to a level where it may be possible to determine interelemental
ratios after subtracting control organism concentrations for the elements
in question. With the present data, the day 28 exposed organism concen-
trations are too close to the control sample data to make reliable
subtractions in all cases except for Cr. Second, a longer uptake
period would give a better evaluation of a long-term effect on the
organism.
89
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Biological Characterization
Solid phase tests
117. Nephtys incisa. In three solid phase tests with N. incisa
there were no mortalities due to Black Rock Harbor (BRH) sediment in 10
days (Tables 13, B1-B3). No worms were seen out of their burrows on
the sediment surface in any treatment. There was a slight difference
in the burrow depth in the different treatments. The depth of the
burrows decreased as the percentage of BRH sediment increased. For
example, in one experiment (Table Bl) the deepest burrow was 1.8 cm in
the 25% BRH treatment but only 0.6 cm in the 100% BRH treatment, and
the estimated average burrow depths in those treatments were 1.2 and
0.4 cm, respectively. The difference in the burrowing depth in the
sediment indicates that the worms are behaving somewhat differently in
the BRH sediment than in the REF sediment, but this parameter is not
particularly useful in short-term sediment characterization tests
because we do not know what a difference in burrow depth means. At
best it can be used as a signal to indicate that there may be some
effect in these treatments in longer term experiments.
118. Neanthes arenaceodentata. In three solid phase tests with
N. arenaceodentata (two with adult males and one with juveniles) there
were no mortalities due to the BRH sediments (Table 13). Control
mortality was 0% in both tests with adults and 5% in the one test with
juveniles (Tables B6-B8). In the tests with adults, one worm from the
100% BRH and one from the 75% BRH treatments came to the surface of
the sediment on day 3 and remained there for the rest of the test.
90
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Table 13
Toxlclty of Solid Phase Black Rock Harbor (Connecticut) Dredged Material
to 11 Species of Marine Invertebrates and Fishes
Species
Annelids
Nephtys
incisa
Heanthes
arenaceodentata
Molluscs
Yoldia limatula
Mulinia lateralis
Arthropods
Test Duration
days
10
10
10
10
LC50*
or
NOEC**
NOEC - 100Z BRH
NOEC - 100Z BRH
NOEC - 66Z BRH
NOEC - 100Z BRH
Behavioral or
Other Effects
Indication of decrease in
burrow depth with BRH mat
Burrowing impaired at 50Z
BRH. No feeding at any
BRH mixture
none
Mysidopsis bahia
Ampellsca abdlta
Fishes
Menldia menidla
Cyprinodon variegatus
Ammodytes americanus
embryo
larvae
Paralichthys
dentatus
Pseudopleuronectes
americanus
4
4
4
4
4
4
4
4
4
NOEC • 100Z BRH
LC50 - 27.2 and 29.8Z
NOEC - 100Z BRH
NOEC - 100Z BRH
NOEC - 100Z BRH
Inconclusive
not definitive
Inconclusive
NOEC - 100Z BRH
Inconclusive
none
Tube building impaired
BRH cone including
none
none
none
none
in all
12.5Z
*LC50 - concentration lethal to an estimated 50% of the test organisms.
**NOEC » no observed effect concentration; effect noted is mortality.
91
-------
These two worms appeared normal otherwise. None of the juveniles
appeared on the surface of the sediment except to search for food.
Because control mortality was 0% in both types of test chambers (indi-
vidual cups and crystallizing dishes), adult If. arenaceodentata do not
have to be exposed in individual cups, if they are given enough sedi-
ment and are not crowded even though these worms are cannibalistic.
Future tests with N. arenaceodentata will be conducted in the crystal-
lizing dishes.
119. Yoldia lima tula. In a preliminary test, Y_. lima tula were
exposed to 25 and 50% BRH sediment and experienced no significant mor-
tality over a 10-day period (Table 13). However, the ability to
burrow into these sediments was impaired (Table 14). These results
indicated that a true 10-day exposure was not produced, so a second
solid phase test was conducted in which, after day 2, the Yoldia in
all treatments were gently pushed into the sediment, the posterior end
down. The animals then burrowed deeper into the sediment. The concen-
trations of BRH sediment and percent mortalities for this test were
as follows: 0% BRH, 0% mortality; 33% BRH, 2.5% mortality; 66% BRH,
7.5% mortality; and 100% BRH, 35% mortality (Table Bll). Even though
exposure times for the two tests are not comparable, neither produced
significant mortalities at or below 66% BRH. The 35% mortality at
100% BRH was statistically significant (Chi-square, P<0.05).
92
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Table 14
Percent of Yoldia not Burrowed into Sediment Over Time for
10
OJ
Treatment Day 1
Solid Phase 1
Re f erence 1.6
25% BRH 45
50% BRH 63.3
Suspended 1
REF Susp/REF Solid 7.5
REF Susp/50% BRH 35
BRH Susp/REF 55
BRH Susp/50% BRH 72.5
Solid Phase Test No. 1 and Suspended Phase Test No. 1
Percent Not Burrowed
2 34567 89
0 00000 00
36,7 26.7 15 11.7 10 13.3 6.7 5
51.7 41.7 33.3 23.3 18.3 15 13.3 11.7
0 00000 00
10 10 5 0 5 0 2.5 0
22.5 17.5 12.5 000 00
40 25 20 10 7.5 5 5 2.5
Day 10
Number Dead/
10 Number Exposed
0 0/61
5 0/60
8.3 1/60
0 0/40
0 3/40
0 0/40
2.5 1/40
-------
120. In addition to the effects on burrowing, the BRH sediment
inhibited Yoldia feeding. This bivalve is a subsurface deposit feeder
which expels unused sediment and feces at the sediment-water interface,
creating a mound of sediment on the surface. These mounds were very
apparent in all control (REF) treatments. In all solid phase BRH expo-
sures, no feeding mounds were seen. Consequently, long-term exposure
may be expected to produce effects on the growth potential of this
bivalve.
121. Mulinia lateralis. There were no lethal or behavioral
effects of BRH sediment on M. lateral!s (Tables 13, B13, and B14).
122. Mysidopsis bahia. The arthropod M. bahia showed no acute
effects following a 96-hr exposure to 100% BRH or REF sediment. The
first test, using exposure cups with screen sides, was monitored daily
for mortality by removing and replacing each cup. This process stirred
up the sediment and created a heavy suspension of particles which made
it very difficult to see the test organisms and also created a suspended
particulate as well as a solid phase test. Although an LC50 for this
test could not be calculated, there was an apparent dose-related
response (Table B16).
123. In order to expose the test species to the solid phase only,
the second and third tests were not checked until day 4. The cups,
therefore, remained undisturbed and no particulate matter was suspended.
There was no 96-hr LCSO with either of these tests and no dose-related
response (Tables 13 and B16-B18).
94
-------
124. Dissolved oxygen in these tests was not affected by the
sediment, and temperature and salinity were within acceptable limits
(Tables 2 and B16-B18).
125. Ampelisca abdita. Table 15 shows 96-hr mortality as a
result of exposures to BRH sediments ranging from 50 to 12.5% mixtures,
with both local (from the Narrow River, R.I. (NR} , local collection
site) sediment and REF sediment as controls. Bioassay methods using
Ampelisca require use of a local sediment for control treatments.
These tests, for each population, are shown in Table 15 for comparison
with the REF sediment. The Long Island (LI) Ampelisca are more sensi-
tive to BRH mixed with local Long Island Sound sediment, than with REF
sediment. The NR Ampelisca experienced no differences in BRH-induced
mortality when exposed to either REF or NR control sediments from
either REF or NR. There was 100% mortality for Narrow River amphipods
to 100% BRH sediment in these and all other preliminary tests. There
was some variability in the quantitative dose response for the six
tests. In all cases, however, 25% BRH produced statistically signif-
icant mortalities (chi-square, P<0.05). The ability to build tubes
was impaired at all BRH exposures and histological analysis showed
tissue damage in the tube-building glands (P.P. Yevich, ERL-N, Personal
Communication).
95
-------
Table 15
Summary of Response Percent Mortality of Ampelisca abdita after 96-hr Exposure in
Solid Phase Tests with Black Rock Harbor Sediment *
Popula-
tion
LI
LI
NR
NR
NR
NR
Mean
size
mm
6.51
4.84
5.53
5.67
5.34
6.80
N
per
cone.
40
40
30
30
80
40
Percent Black Rock
Harbor Sediment
Control
Sediment
REF
LI
REF
REF
NR
REF
Test #
1
2
3
4
5
6
50%
33.0
74.8
83.3
80.0
84.8
61.0
25% 12.5%
25.0
56.1
63.3
63.3
45.0 4.0
49.1 30.6
Control
7.5
10.9
3.3
0.0
1.8
11,6
LC50
% BRH
27.16
29.8
* Amphipods were collected-from Narrow River (NR) and Long Island Sound (LI). Control sediments for
Black Rock Harbor mixtures were collected from Narrow River (NR), Long Island Sound (LI), and
Reference Station South (REF).
-------
126. Menidia menidia. Larvae of M. menidia showed no acute
effects following a 96-hr exposure to 100% BRH or REF sediment. No
mortality occurred in any treatment (SW control; 100% REF; 100% BRH)
for either of the two tests conducted using the solid phase test regime
(Tables B29 and B30). No obvious behavioral changes were observed in
swimming behavior at the end of the test period, and all fish appeared
to be feeding on the brine shrimp supplied. Dissolved oxygen concen-
trations were close to saturation for all treatments.
127. Cyprinodon variegatus. Larvae of £. variegatus appeared
to be unaffected by the solid phase of either REF or BRH. A single
mortality occurred in one seawater control of the first test (Table
B33), Dissolved oxygen was at or above saturation. Mortality in the
control appeared to be the result of gas supersaturation. In the
second test, the dissolved oxygen was close to saturation in all treat-
ments (Table B34).
128. Ammodytes americanus. Embryos of A_. americanus appeared
to be unaffected by the solid phases of both REF and BRH. Mortalities
occurred, but not in response to dose (Tables B37 and B38).
129. Ammodytes americanus larval solid phase tests showed high
mortality at 100% BRH when three treatments (SW, REF, and 100% BRH)
were used (Tables B39 and B40). A chi-square test showed significant
differences between mortalities in the 100% REF and 100% BRH (P<0.01).
Subsequent testing with six treatments showed high control mortality
(100% REF) as well as high mortality across the BRH treatments (Tables
B41-B44). These tests generally showed higher mortality at higher
97
-------
concentrations of BRH sediment, but test and probit analysis (Finney
1971) results were inconclusive.
130. Ammodytes americanus has not been used extensively in labo-
ratory tests and the high mortality in these tests can probably be
attributed to handling stress. Further testing of this species is
required to find the optimum method of handling, the optimum age for
testing, and the most suitable bioassay chamber to minimize control
mortality. Dissolved oxygen measurements were taken only once during
all the solid phase tests. Dissolved oxygen was assumed to be at
saturation, but the measurements taken at the time of the last test
(Table B44) were below saturation, and could have contributed to the
high mortality observed during these tests.
131. Paralichthys dentatus. The fish P_. dentata appeared to be
unaffected by the presence of either REF or BRH in the solid phase.
Control and REF mortalities were within accepted ASTM limits «10%)
for both tests. Dissolved oxygen was at or near saturation in all the
exposure chambers (Tables B47 and B48).
132. Pseudopleuronectes americanus. The results of the solid
phase tests with winter flounder larvae were not conclusive. In test
1 there was significantly (chi-square test, P<0.05) higher mortality
in REF than there was in BRH. In the second test there was significantly
(chi-square test, P<0.05) higher mortality in BRH than there was in
REF. In both tests there was a tendency for the larvae in the exposure
chambers containing both REF and BRH sediment to stay in the water
column more of the time than the larvae in the seawater controls. The
98
-------
dissolved oxygen concentration was at or near saturation in all treat-
ments in both tests (Tables B49 and B50).
Suspended particulate tests
133. Nephtys incisa. In two suspended particulate tests with
Nephtys iricisa no toxic effect was seen (Table B4 and B5). Mortality
was 5% or less in all treatments. During the test, no worms appeared on
the sediment surface. There were no differences in the estimated average
depth of burrows in the various treatments (Table 16).
134. In two earlier suspended particulate tests with _N. incisa a
toxic effect (25 and 20% mortality) was seen in the 25 mg/L BRH sus-
pended particulate, 100% BRH solid phase treatment. However, in a
review of the procedures for these two tests, we discovered that we
had not turned off the inflow water when we added the worms to the treat-
ment dishes. When we repeated the tests (above), we saw no toxic effects
in any treatments. We concluded that the "mortalities" in the first two
tests were not real and probably resulted from the worms not burrowing
into the sediment in the beginning of the test. Therefore, it is impor-
tant that the procedures outlined in the methods section be followed
carefully to avoid this problem.
135. Neanthes arenaceodenta. In the suspended particulate tests with
,N. arenaceodentata, no toxic effect was seen in any treatment (mortality
10% or less, Table B9a and B9b). No worms appeared on the surface in any
treatments except to search for food (a normal activity).
99
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Table 16
Toxicity of Black Rock Harbor (Connecticut) Dredged Material, as Suspended Sediment,
to 10 Species of Marine Invertebrates and Fishes
o
o
Species
Annelids
Nephtys
incisa
Neanthes
arenaceodentata
Molluscs
Yoldia limatula
Mulinia lateral!s
Arthropods
Mysidopsis bahia
Ampelisca abdita
Fishes
Menidia menidia
Cyprinodon variegatus
Ammodytes americanus
Pseudopleuronectes
americanus
Test Duration
days
10
10
10
10
4
4
LC50*
or
NOEO*
NOEC = 100% solid and
25 mg/L suspended BRH material
NOEC = 100% solid and 25 mg/L
suspended BRH material
NOEC = 100% solid and 40 mg/L
suspended BRH material
NOEC = 100% solid and 40 mg/L
suspended BRH material
NOEC = 100% solid and 25 mg/L
suspended BRH material
NOEC - 0% solid and 20 mg/L
suspended BRH material
NOEC = 100% solid and 25 mg/L
suspended BRH material
NOEC - 100% solid and 25 mg/L
suspended BRH material
Not definitive
NOEC = 100% solid and 25 mg/L
suspended BRH material
Behavioral or
Other Effects
None
None
No burrowing in 50%
BRH solid phase mat
None
None
None
None
None
None
*LC50 = concentration lethal to an estimated 50% of the test organisms.
**NOEC = no observed effect concentration, effect noted is mortality.
-------
136. Yoldia limatula. There were no mortalities with Ypldia
exposed to 40 mg/L BRH suspension with either REF or 50% BRH sediment
as a solid phase (Tables BIO to B12). In the treatments with 50% BRH
as a solid phase, the same nonburrowing response that was noted in the
solid phase tests was seen.
137. Mulinia lateralis. Mulinia experienced no mortalities when
exposed to 40 mg/L BRH suspended participates with 100% BRH in the solid
phase (Tables B13 to B15).
138. Mysidopsis bahia. The arthropod M. bahia showed no acute
effects from a 96-hr exposure to 25 mg/L BRH or REF participate. The
two 96-hr tests for the particulate phase were conducted at 21°C rather
than 25°C as in the solid phase tests, however; the lower temperature
had no effect on the survival of the test species.
139. In the first test, the control was without reference sedi-
ment; in the second test the control contained reference sediment.
No difference in mortality occurred between either of these two treat-
ments or between particulate phase controls and the controls from the
solid phase tests.
140. The levels of exposure for the particulate phase tests were
representative of the worst case situation with the highest level of
BRH particulate load (25 mg/L) and 100% BRH sediment compared to the
identical condition with reference particulate and sediment. Neither
test showed any effect on survival at 96 hours (Tables 16, B19, and
B20).
101
-------
141. Dissolved oxygen, temperature, and salinity remained within
acceptable limits throughout the tests (Tables 2, B19, and B20), Elec-
tronic particle counts showed consistency tor particle density in the
splitters, exposure chambers, and test chambers.
142. Ampelisca abdita. The species A_. abdita experienced no sig-
nificant mortalities when exposed to 20 mg/L BRH at 20°C (Table B27)
or when exposed to 25 mg/L BRH at 8°C (Table B28). In both cases, a
no-effect level of 0% BRH was used in the solid phase (Table 16).
143. Menidia menidia. Larvae of bl. menidia showed no significant
acute effects following a 96-hr exposure to suspended particulates of
BRH or REF sediment (Table 16). A single mortality occurred in one SW
control dish (Table B31). Although it was difficult to observe fish
swimming in the REF and BRH exposure dishes, it could be discerned
during daily counts that fish were feeding on the brine shrimp ration.
Larvae appeared to be swimming normally at the termination of these
experiments. Dissolved oxygen was close to saturation for all treat-
ments.
144. Cyprinodon variegatus. The fish C_. variegatus appeared to
be unaffected by exposure to either the BRH or the REF 'sediments in
suspended phase at the nominal concentration of 25 mg/L (Table 16).
There were no mortalities in any of the treatments in either test, and
the behavior of the larvae appeared to be unaffected by the suspended
sediment. Dissolved oxygen was close to saturation in all treatments
(Tables B35 and B36).
102
-------
145. Ammodytes americanus. Suspended sediment test results for
A. americanus were inconclusive as to the effect of the suspensions of
100% REF or BRH on the larvae (Tables B45 and B46).
146. Psuedopleuronectes americanus^ Larvae of P. americanus did
not appear to be adversely affected by the presence of suspended REF or
BRH at the nominal concentration of 25 mg/L (Table 16). In the first
test there were only two mortalities, one in the seawater control and
one in BRH (Tables B51 and B52). In the second test, there were some
mortalities in all of the treatments (5 out of 24 in the seawater con-
trol, 5 out of 19 in REF, and 4 out of 22 in BRH), but there was no
significant difference between the REF and the BRH in either test.
Dissolved oxygen was at or near saturation in all exposure dishes
during both tests. As in the solid phase tests with the P. americanus,
there was more of a tendency for the larvae to stay in the water column
in the exposure chambers containing sediment than in those containing
no sediment.
103
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PART VI: CONCLUSIONS AND RECOMMENDATIONS
147. Black Rock Harbor dredged material contained substantial
concentrations of both organic and inorganic contaminants that were
biologically available to the blue mussel Mytilus edulis in a suspended
particulate bioaccumulation study. PCB's were present in the sediment
at 6800 ng/g (ppb), while PAH's of molecular weights between 128 and
302 were present at concentrations between 17 and 9100 ng/g. Alkyl
homologs of the PAH's of molecular weights between 128 and 252 were
also present in the sediment at concentrations between 110 and 13000
ng/g, measured as the sum of alkyl homologs between C-l and C-4. Of
the organic contaminants present in the sediment, mussels accumulated
PCB's to a tissue concentration of 3000 ng/g, which is 44% of the con-
centration in the sediment. PAH's of molecular weights 202, 228, and
252 were accumulated to tissue concentrations between 1600 and 2000 ng/g,
which are between 18 and 28% of the concentrations in the sediment. Other
PAH's were accumulated to lesser extents. Alkyl homologs of the PAH's
were also accumulated, with a maximum tissue concentration of 5000 ng/g
for the 178 alkyl homologs, which is 38% of the sediment concentration.
Other alkyl homologs from molecular weight PAH's of 166 through 228 were
accumulated to levels between 110 and 3400 ng/g; these tissue concentra-
tions are between 14 and 41% of the sediment concentration. Although
these concentrations of contaminants accumulated in mussels were after a
28 day exposure, the concentrations in the mussels after only 7 days were
close to the same concentrations for many of the contaminants.
104
-------
148. Inorganic contaminants were also present in Black Rock
Harbor sediment, but an interference precluded the measurement of As.
With the exception of Mn, Zn, As, and Hg, which were below the limit
of detection, all other trace metals showed statistically significant
(P<.05) increases over controls during the 28 day mussel uptake study.
The greatest uptake was for Cr, which reached a concentration of 25 yg/g
at the end of the bioaccuraulation compared to a control concentration
of 1.96 yg/g. Other trace metals were taken up to a lesser degree,
including Cd which reached a concentration of 7 pg/g compared to a
control value of 2.8 yg/g. Compared to the sediment concentration,
the organisms accumulated to only 2% of the Cr, but 28% of the Cd
sediment concentration values.
149. In contrast to the organic contaminants which reached high
concentrations relatively early in the bioaccumulation study, many trace
metals had not reached plateau concentrations by the end of the 28-day
study. Therefore, although many organic contaminants should be detected
in a shorter test, significant accumulation from inorganic contaminants
could be overlooked.
150. Black Rock Harbor material as solid phase or in combination
with the suspended particulate phase was acutely lethal to one of the
eleven species tested and caused behavioral changes in two species.
Only with Ampelisca abdita was the material sufficiently toxic to pro-
duce 96-hr LC50 values (27.2 and 28.2% BRH, solid phase; Tables B25
and B26). In addition, the amphipod's tube building was impaired in
all BRH concentration down to 12.5%, the lowest concentration tested.
105
-------
Yoldia limatula failed to burrow into sediment containing 25% BRH or
higher and did not feed even when gently pushed into the sediments.
151. All species except Ammodytes americanus proved suitable
for testing dredged material in this study. As stated earlier, .A.
americanus has never before (to our knowledge) been used in aquatic
toxicology and much must be learned in the handling and culture of the
species before acceptable control survival «10%; ASTM 1980b) can be
attained. A strong effort will continue in developing the species for
toxicity testing because it is an important link in marine food chains
in coastal waters of the northeastern United States (Sherman et al.
1981) and may be impacted by dredged material disposal.
152. Three of the five infaunal species tested (A. abdita, N.
incisa, and Y_. lima tula) were sensitive to BRH material in acute tests,
whereas no epibenthic or water column species showed sensitivity to
the material either in solid phase or in combination with the suspended
participate phase. For the affected species, the most important factor
was the concentration (or presence) of BRH solid phase material.
Without further data from long-term tests, including measurements of
energetics, histopathology, etc., one can only speculate on the reason
for the differing sensitivities, but it may be related to whether or
not the BRH solid phase material is in contact with the species. For
example, larval A. americanus and VL. menidia seldom directly contact
the benthos. Conversely, infaunal species such as If. lima tula and A_.
abdita are in intimate contact with the sediment, and thus the toxic
properties of the BRH material are available by direct contact or by
106
-------
ingestion. Longer term exposures and more detailed studies may reveal
the factors causing the different sensitivities.
153. The reproducibility of the effects observed in the solid
phase and in combination with the suspended particulate phase for the
species tested was very good. No significant differences were observed
in replicate solid phase tests with JN. incisa, N. arenaceodentata, M.
lateralis, JM. bahia, A. abdita, M. menidia, ^. variegatus, P_. dentata,
and P^. americanus. As stated earlier, further research is needed with
A_. americanus to obtain the necessary control survival and test repro-
ducibility; Y_. limatula tests were not precisely replicated (Tables
BIO and Bll), but the mortality observed in the two tests was corre-
lated well with BRH concentration.
154. Replicates from suspended particulate tests, like the solid
phase replicates, showed excellent reproducibility. No significant
differences in results of the replicates were observed in N. incisa,
M^ lateralis, M^ bahia, M. menidia, CL variegatus, and JP. americanus.
Suspended particulate test results obviously depended upon the reli-
ability of the dosing system. Once the initial problems were solved,
the microprocessor, transmissometer, and dosing valve system worked
well. Generally, the particulate concentration was maintained within
10% of the desired values.
107
-------
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110
-------
APPENDIX A: CHEMICAL DATA
Appendix Tables A1-A7 list the concentration of each polynuclear
aromatic hydrocarbon and its alkylated homolog from C-l through C-4
for the three replicates of day 7, 14, and 28 and for the three repli-
cates of both the day 0 and day 28 controls. Also given are the same
measurements for Black Rock Harbor sediment and for the analytical
blank associated with these analyses.
Appendix Tables A8-A11 give complete metal concentration data for
all sediment and mussel samples.
Al
-------
Table Al
Day 0 control concentrations of PAH and Aklyl Homologs
for replicates A, B, and
Concentration, ng/g
Compound
128
154Bi
154An
166
178
202
228
252
276
278
300
302
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
PAH
1.2
0.5
0.8
1.6
1.3
1.1
0.9
0.3
1.0
1.8
1.1
1.3
9.1
6.1
6.5
42
27
34
9.7
5.9
10
17
11
18
4.0
2.7
3.2
<0.4
<0.2
<0.6
<0.1
<0.1
<0.1
<0.3
<0.5
<0.2
C-l
0.6
0.3
0.7
0.9
0.5
0.5
_
-
—
1.0
0.6
1.0
11
7.4
8.6
8.7
5.4
7.7
2.0
1.3
3.2
<6.6
6.3
<5.9
C-2
2.3
1.3
2.2
2.1
1.6
2.4
1.4
0.8
1.6
4.6
2.7
3.7
19
12
15
2.7
3.0
3.6
1.5
1.6
2.3
<0.7
<0.4
<0.2
C
Dry Weight
C-3
4.4
2.6
2.4
3.8
2.2
3.6
2.0
1.1
2.1
9.2
5.6
7.0
11
8.1
11
1.5
1.1
1.5
2.6
1.4
1.9
<0.3
<0.2
<0.1
C-4
4.4
1.8
2.8
4.5
2.9
4.2
0
-
—
3.2
2.4
2.4
3.2
3.3
3.4
0.6
0.6
0.4
_
-
—
<0.1
<0.1
<0.1
A2
-------
Table A2
Day 7 exposed concentrations of PAH and Alkyl Homologs
for replicates A, B, and C
Concentration, ng/g Dry Weight
Compound
128
154Bi
154An
166
17&
202
228
252
276
278
300
302
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
PAH
0.4
0.5
0.6
1.5
1.6
2.5
2.9
4.4
3.6
14
13
15
190
240
220
230
3100
2300
1700
2100
1700
1200
1600
1200
220
330
210
120
170
130
9.4
14
9.0
57
81
60
C-l
0.5
0.6
0.5
6.8
7.0
7,4
15
23
19
72
71
66
930
1200
970
1600
2200
1600
830
1100
800
300
410
290
C-2
7.1
15
9.1
70
79
70
76
110
93
260
270
250
1900
2400
1800
950
1200
900
380
480
320
110
150
100
C-3
80
170
120
200
240
200
160
200
190
430
490
370
1500
2000
1400
490
640
460
140
180
110
36
53
39
'
C-4
190
330
250
380
490
280
_
-
-
340
370
260
870
1200
800
170
250
160
49
65
40
10
14
13
A3
-------
Table A3
Day 14 exposed concentrations of PAH and Alkyl Homologs
for replicates A, B, and C
Concentration, ng/g Dry Weight
Compound
128
154B1
154An
166
178
202
228
252
276
278
300
302
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
b
C
A
B
C
PAH
0.8
1.0
1.4
2.3
2.4
4.1
3.4
5.3
5.5
6.8
8.8
9.3
120
160
150
930
1200
1200
660
950
910
490
680
650
97
140
110
52
85
67
2.1
3.8
2.7
12
22
16
C-l
0.5
1.3
1.6
2.6
4.6
3.9
18
-
—
33
52
44
390
530
480
630
850
840
340
520
490
140
190
150
C-2
8.4
16
16
28
43
40
42
63
63
100
150
130
720
1000
980
360
540
510
140
220
180
38
65
49
C-3
65
110
96
89
140
120
84
110
110
170
230
230
590
890
810
180
300
260
49
76
57
8.3
18
12
C-4
120
190
180
170
400
300
-
-
-*
120
200
160
300
540
440
80
99
72
16
29
20
<0.9
<4.0
<2.2
A4
-------
Table A4
Day 28 Exposed concentrations of PAH and Alkyl Homologs
for replicates A, ft, and C
Concentration, ng/g Dry Weight
Compound
128
154B1
154An
166
178
202
228
252
276
278
300
302
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
PAH
0.5
0.4
1.0
2.8
1.8
5.8
3.8
2.0
5.6
13
7.9
15
210
140
180
2500
1600
1800
2200
1500
1700
1800
1400
1500
330
230
280
180
140
160
9.6
8.4
9.9
62
46
60
C-l
0.8
0.3
1.4
7.0
5.0
7.5
21
13
41
67
44
62
1000
650
730
1900
1300
1400
1300
970
1000
450
350
400
*
C-2
10
4.4
15
78
46
71
100
54
98
290
180
220
2300
1400
1400
1300
910
940
570
440
450
160
120
140
C-3
100
53
130
250
140
200
210
120
180
520
310
590
2100
1300
1300
750
550
560
200
140
170
47
31
45
C-4
250
130
220
620
300
430
_
-
-
420
270
290
1300
810
840
280
210
210
58
49
46
12
10
10
A5
-------
Table A5
Day 28 control concentrations of PAH and Alkyl Homologs
for replicates A, B, and C
Concentration, ng/g Dry Weight
Compound
128
154Bi
154An
166
178
202
228
252
276
278
300
302
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
C
A
B
A
B
C
A
B
C
A
B
C
A
B
C
PAH
0.5
0.5
0.6
6.5
3.0
6.3
1.0
1.1
1.4
1.4
0.8
1.4
6.8
5.1
8.1
22
17
19
4.3
2.4
2.5
7.8
4.8
2.2
<1.0
0.9
<0.9
<0.3
0.6
<0.1
<0.1
<0.1
<0.1
<0.1
<0.1
C-l
0.6
0.6
0.7
3.6
1.8
3.4
_
-
-
2.6
1.7
2.4
14
12
15
5.3
3.3
4.4
1.9
1.0
0.9 '
<4.1
<3.0
C-2
2.3
1.6
2.5
5.5
5.5
4.6
3.6
3.4
4.0
7.6
6.0
7.5
16
11
14
3.1
1.3
1.1
<1.8
<0.8
<1.1
<0.5
<0.1
C-3
4.7
4.0
5.5
9.0
7.2
8.7
5.0
3.5
3.8
11
7.9
9.3
11
6.2
7.6
2.1
0.9
0.7
<1.4
<0.9
<1.4
<0.2
<0.1
C-4
9.5
7.6
9.6
12
9.0
12
_
-
—
6.6
2.5
4.1
4.9
2.5
3.3
0.5
<0.3
0.5
<5.5
<4.1
<7.0
<0.1
<0.1
A6
-------
Table A6
Analytical blank concentrations of PAH and Alkyl Homologs
Compound
128
154Bi
154An
166
178
202
228
252
276
278
300
302
of previous samples
Concentration, ng/g Dry Weight
PAH C-l C-2 C-3 C-4
0.6 0.3 <0.8 0.5 <0.4
0.7 <0.2 <0.3 <0.4 <0.3
0.6 - <0.1 <0.1
0.2 2.5 <0.3 <1.3 <0.1
0.6 0.5 <0.4 <0.3 <0.2
0.7 <0.1 <0.1 <0.1 <0.1
0.7 <0.1 <0.2 <1.0 <0.1
0.7 <2.4 <0.1 <0.1 <0.1
<0.1
<0.1
<0.1
<0.1
A7
-------
Table A7
concentrations in ng/g dry weight
Concentration, ng/g Dry Weight
Compound
128
154Bi
154An
166
178
202
228
252
276
278
300
302
PAH
17
54
120
370
2700
7100
9800
8600
9100
4400
130
2700
C-l
67
130
140
610
3500
3800
5400
2700
C-2 C-3
420 1200
520 760
420 500
930 1100
3900 3200
2200 1500
3900 2200
1500 580
C-4
1700
1500
-
790
2300
770
1200
-
AS
-------
Table AS
Metal Concentrations for Black Rock Harbor Barrel 100.
All Sediment Samples were
Dissolved with Hot
Concentrated HNOq.
All Concentrations are in ug/g Dry Weight
TOP
1
2
3
AVE
SD
ZSD
MIDDLE
1
2
3
AVE
SD
%SD
BOTTOM
1
2
3 -
AVE
SD
ZSD
BARREL
AVE
SD
ZSD
BLANK
Fe
31000
28400
28900
29400
1130
3
30400
30100
29800
30100
244
1
29400
28700
30100
29400
571
1
29600
809
2
1 0.
BLANK 2 0.
Mn
403
369
303
326
52
16
326
417
384
375
37
10
348
371
408
375
24
6
359
37
10
90 0.025
84 0.014
Zn
1190
1090
1150
1130
41
3
1290
1280
1150
1240
63
5
1210
1200
1220
1210
8
1
1200
59
4
0.34
0.39
Cu
2350
2140
2390
2293
109
4
2480
2490
2270
2413
101
4
2410
2420
2520
2450
49
2
2385
112
4
0.25
0.34
Pb
384
346
365
365
15
4
403
383
364
383
15
4
381
380
400
387
9
2
378
16
4
0.14
0.17
Cd
24.5
23.5
23.8
23.9
0.4
1.8
23.2
24.1
22.2
23.2
0.8
3.4
23.8
21.7
23.9
23.1
1.0
4.4
23.4
0.9
3.7
0.056
0.056
Cr
1400
1260
1380
1346
61
4
1540
1460
1410
1470
53
3
1460
1460
1510
1476
23
1
1431
77
5
0.014
0.017
Ni
141
133
144
139
4
3
144
147
139
143
3
2
131
139
141
137
4
3
139
4
3
0.20
0.56
Hg
1.8
1.6
1.7
1.7
0.1
7.0
1.7
1.7
1.6
1.7
0.1
4.0
1.7
1.7
1.8
1.7
0.1
1.0
1.7
0.1
4.0
<0.01
<0.01
A9
-------
Table A9
Metal Concentrations for Black Rock Harbor Barrel #LL.
All Sediment Samples were Eluted with 5Z HNO^.
All Concentrations are Given in ug/g Dry Weight
TOP
1
2
3
AVE
SD
%SD
MIDDLE
1
2
3
AVE
SD
%SD
BOTTOM
1
2
3
AVE
SD
%SD
BARREL
AVE
SD
%SD
BLANK
BLANK
Fe
26400
26500
27200
26700
355
1
26300
28000
26700
27000
712
2
26100
26200
25800
26000
169
1
26600
623
2
1 1.9
2 1.3
Mn
284
278
284
282
2
1
274
280
286
280
4
1
269
272
278
273
3
1
278
5
1
0.07
0.03
Zn
1290
1210
1240
1250
33
2
1180
1282
1244
1240
42
3
1150
1180
1150
1160
14
1
1210
50
4
1.
0.
Cu
2560
2510
2650
2570
57
2
2500
2670
2510
2560
77
3
2530
2490
2490
2500
18
1
2540
64
2
19 1.03
32 0.71
Pb
421
431
427
426
4
1
436
4.16
377
409
24
5
393
397
425
405
14
3
413
18
4
0.
0.
Cd
25.4
24.8
25.7
25.3
0.4
1.5
24.0
25.6
24.1
24.6
0.7
3.0
23.5
24.4
24.9
24.3
0.6
2.4
24.7
0.7
2.9
055 0.032
063 0.048
Cr
1380
1350
1350
1360
14
1
1330
1380
1360
1360
20
1
1350
1280
1310
1310
28
' 2
1340
30
2
0.048
0.024
Nl
172
166
168
168
2
1
168
184
170
174
7
4
164
164
167
165
1
1
169
5
3
0.79
0.95
A10
-------
Table A10
Metal Concentrations Determined for Mytilus edulis
Control Samples and Black Rock
Harbor Exposed Samples.
All Concentrations in ug/gram Dry Weight
SAMPLE
DAY 1
7 2
3
AVE
SD
%SD
DAY 1
14 2
3
AVE
SD
%SD
DAY 1
28 2
3
AVE
SD
ZSD
Fe
404
334
332
357
41
11
322
333
333
329
6
2
715
348
437
500
191
38
Mn
15.6
9.5
11.3
12.1
3.2
25
19.7
38.9
24.9
31.1
10.1
32
16.8
7.6
9.2
11.2
4.9
44
Zn
350
236
272
286
58
20
176
163
142
160
17
11
386
236
376
333
84
25
Cu
81.3
68.6
36.3
62.0
23.2
37
68.6
73.6
83.3
75.2
7.4
10
74.9
39.9
50.2
55.0
18.0
33
Pb
11.9
11.2
8.9
10.7
1.5
14
9.50
8.20
7.87
8.52
0.86
10
19.0
9.8
13.0
13.9
4.7
34
Cd
5.37
4.71
4.00
4.69
0.69
15
3.67
2.73
2.93
3.11
0.49
16
8.09
4.72
8.09
6.97
1.94
28
Cr
17.2
14.6
14.2
15.3
1.6
10
13.3
10.8
12.0
12.0
1.3
10
36.4
15.1
23.6
25.1
10.7
43
As
9.37
7.43
8.95
8.58
1.02
12
6.94
7.33
7.67
9.31
0.37
5
9.86
7.76
9.00
8.87
1.06
12
(continued)
All
-------
Table AID (Cont'd)
CONTROL SAMPLES
SAMPLE
DAY 1
0 2
3
AVE
SD
%SD
DAY 1
28 2
3
AVE
SD
%SD
Fe
189
178
229
199
26
13
220
212
206
213
7
3
Mn
11.4
10.4
11.6
11.1
0.7
6
9.3
8.1
21.0
12.8
7.1
55
Zn
70
154
223
149
76
51
289
164
210
221
63
29
Cu
51.8
10.1
14.2
25.4
23.0
91
20.9
19.7
11.8
17.4
5.0
28
Pb
4.73
4.85
4.57
4.72
0.14
3
9.42
5.81
4.40
6.54
2.59
40
Cd
3.09
2.54
2.50
2.71
0.32
12
3.32
2.69
2.35
2.79
0.49
18
Cr
2.64
2.08
2.07
2.26
0.32
14
2.53
1.81
1.55
1.96
0.50
26
As
7.45
8.62
8.50
8.19
0.64
8
7.37
8.38
6.45
7.40
0.96
13
BLANK SAMPLES
AVE <1.9 <0.06 <0.6 >0.3 <0.06 <0.01 <0.06 <0.2
A12
-------
Table All
Percent Recovery of Metals Spiked Into Mussel Samples
Element
Fe
Zn
Mn
Cu
Pb
Cd
Cr
As
yg Added
15.0
62.5
25.0
37.5
2.50
2.50
2.50
2.50
1.25
2.50
0.625
0.625
2.50
2.50
2.50
2.50
yg Found
14.5
59.8
25.2
38.5
2.66
2.95
2.31
2.38
1.07
2.34
0.630
0.66U
2.43
2.44
2.28
2.17
% Recovery
96.7
95.6
101
103
106
118
92.4
95.2
85.6
93.6
101
106
97.2
97.6
91.2
86.8
A13
------- |