UPTAKE  DEnillATn,
'
                               FIELD VERIFICATION PROGRAM
                                   (AQUATIC DISPOSAL)


                                  TECHNICAL REPORT D-85-2


                   BIOACCUMULATION  OF  CONTAMINANTS  FROM
                     BLACK ROCK  HARBOR  DREDGED  MATERIAL
                          BY MUSSELS AND POLYCHAETES

                                           by
                       James Lake,  Gerald L. Hoffman. Steven C. Schimmei

                              Environmental Research Laboratory
                              US Environmental Protection Agency
                              Narragansett. Rhode Island 02882
February 1985
 Final Report
                              Approved For Pub'
                            prepared tor DEPARTMENT OF THE ARMY
                                US Army Corps of Engineers
                                Washington, DC  20314-1000
                            and  US Environmental Protection Agency
                                  Washington, DC  20460
                                 r^ 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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           Unclassified
SECURITY CLASSIFICATION OF THIS PAGE (Wim r>nta Kutarnd)
            REPORT DOCUMENTATION PAGE
                                               READ INSTRUCTIONS
                                            BKI'OKE COMPUKTINO KORM
 1. REPORT NUMBER
  Technical Report D-85-2
                                      2. GOVT ACCESSION NO
                                                        3. RECIPIENT'S CATALOG NUMBER
 4. TITLE (and Subtitle)
  BIOACCUMULATION OF CONTAMINANTS FROM BLACK
  ROCK HARBOR  DREDGED MATERIAL BY MUSSELS AND
  POLYCHAETES
                                       5. TYPE OF REPORT a PERIOD COVERED

                                        Final report
                                       6. PERFORMING ORG. REPORT NUMBER
 7. AUTHORf")
  James Lake,  Gerald L. Hoffman,
  Steven C.  Schimmel
                                                        8. CONTRACT OR GRANT NUMBER(«)
 9. PERFORMING ORGANIZATION NAME AND ADDRESS
  US Environmental Protection Agency
  Environmental  Research Laboratory
  Narragansett,  Rhode Island  02882
                                       10. PROGRAM ELEMENT, PROJECT, TASK
                                          AREA a WORK UNIT NUMBERS
                                         Field Verification Program
                                         (Aquatic Disposal)
 II. 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
                                         February 1985
                                       13. NUMBER OF PAGES
                                         150
 T4.  MONITORING AGENCY NAME « ADORESSf// dlllerent from Controlling Olllce)
  US Army Engineer Waterways Experiment Station
  Environmental  Laboratory
  PO Box 631, Vicksburg, Mississippi  39180-0631
                                       15. SECURITY CLASS, (at thle report)
                                        Unclassified
                                       ISa.
DECLASSIFI CATION/ DOWN GRADING
SCHEDULE
 16. DISTRIBUTION STATEMEN T (at thle Report)
  Approved for public release;  distribution unlimited.
 17. DISTRIBUTION STATEMENT (of the abstract entered In Block 30, It different from Report)
 18. SUPPLEMENTARY NOTES
  Available from  National Technical Information  Service, 5285  Port Royal Road,
  Springfield, Virginia  22161.  Appendices A and  B are on microfiche and are
  enclosed in a pocket attached  to  the back cover.
19. KEY WORDS (Continue on reveree title it neceateiy end Identify by block number)
 Absorption (Physiology)   (LC)
 Dredged  materials—research   (LC)
 Marine pollution—measurement  (LC)
 Polychlorinated biphenyls  (LC)
20. ABSTRACT (Continue an rerere* aMb H n*c+eemr? fad Identity by block number)
        Mussels  (Mytilus edulis)  and worms  (Nereis  virens) were  exposed in
  laboratory studies to dredged material from Black Rock Harbor  (BRH),
  Connecticut, to examine the bioaccumulation of  organic and inorganic contam-
  inants.  Mussels were exposed in  a dosing system  designed to maintain a
  constant concentration of suspended particulates  and food (algae)  in seawater.
  Control mussels received only food (algae).  Monitoring of concentrations
                                                            (Continued)
DO ,^1473
EDITION OF I MOV S» IS OBSOLETE
                                         Unclassified
                                            SECURITY CLASSIFICATION OF THIS PAriE (When Data Entered)

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  	Unclassified	
SECURITY CLASSIFICATION OF THIS PAGE(H7l«n Dflf Entarfd)
 20.  ABSTRACT (Continued).

 of organic and inorganic contaminants showed that the system maintained
 constant concentrations during exposure.  Exposed mussels accumulated organic
 compounds and some inorganic elements, reaching steady-state values between  the
 first and second weeks of exposure.  During the 28-day exposure period, mussels
 showed increases in concentration of two to three orders of magnitude for
 organic contaminants, but those metals accumulated showed increases of less
 than a factor of 12.
       In general, the depuration of organic contaminants was rapid during the
 first week of depuration, and the depuration rate was inversely related to
 the compound's n-octanol/water partition coefficient.  After the first week
 depuration rates decreased, and concentrations of most organic compounds re-
 mained above control values to the end of the 5-week depuration period.  Iron
 and chromium depurated to control levels within a 2-week period.
       The polychaete worm ^N. virens was exposed to BRH bedded sediment in glass
 aquaria maintained under flowing seawater.  Other worms were maintained in
 reference sediments.  Worms exposed for 28 days accumulated polychlorinated
 biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) to concentrations
 one to three orders of magnitude above those found in the reference organisms.
 Of the metals determined, only Cr and Cu were found to accumulate to concen-
 trations higher than those in the reference worms.  Concentrations of PCBs
 exposed to BRH sediment did not decrease during a 28-day depuration period in
 reference sediments, but depuration of PAHs was apparent.  Chromium and copper
 depurated to control levels after 2 weeks.
       Bioaccumulation factors for PCBs calculated for mussels and worms, when
 total exposure concentrations were normalized to a gram dry weight sediment
 basis, were generally within a factor of 1.5.  This suggests that modeling
 bioaccumulation of some organic compounds as a partitioning of contaminants
 between sediments and organisms may have promise as a generalized predictive
 technique.
                                                    Unclassified
                                        SECURITY CLASSIFICATION OF THIS PAGEf»7i»n Dfta Entered)

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SUBJECT:  Transmittal of Field Verification Program Technical Report Entitled
          "Bioaccumulation of Contaminants from Black Rock Harbor Dredged
          Material by Mussels and Polychaetes"

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 (ERLN) in Narragansett, RI, made it clear
that a dredging project at Black Rock Harbor in Bridgeport, CT, presented a
unique opportunity for simultaneous evaluation of aquatic disposal, upland
disposal, and wetland creation using the same dredged material.  Evaluations
were to be based on technology existing within the two agencies or developed
during the six-year life of the program.

3.  The program is generic in nature and will provide techniques and 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:  Transmlttal of Field Verification Program Technical Report Entitled
          "Bioaccumulation of Contaminants from Black Rock Harbor Dredged
          Material by Mussels and Polychaetes"
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.
      ChoromokosV Jr77~?fc.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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                                    PREFACE






      This report describes work performed by the U.S. Environmental Protec-




tion Agency (EPA), Environmental Research Laboratory, Narragansett, R.I.




(ERLN), as part of the Interagency Field Verification of Testing and Predic-




tive Methodologies for Dredged Material Disposal Alternatives Program or the



Field Verification Program (FVP).   The FVP,  sponsored by the Office, Chief of




Engineers (OCE), is assigned to the Environmental Laboratory (EL),  U. S. Army




Engineer Waterways Experiment Station (WES),  and is managed under the Environ-




mental Effects of Dredging Programs (EEDP).   The OCE Technical Monitors for




FVP were Dr. John R. Hall and Dr.  William L.  Klesch.




      The objective of the FVP is to verify existing predictive techniques for




evaluating the environmental consequence of dredged material disposal under




aquatic, wetland, and upland conditions.  The aquatic portion of this study is




being conducted by ERLN,  with the wetland and upland portions conducted by WES.



      The principal ERLN investigators for this aquatic study were Drs. James




Lake and Gerald Hoffman,  Analytical Chemists, and Mr. Steven Schimmel, Aquatic




Toxicologist.   Laboratory exposure system design was coordinated by Mr. Jay




Sinnett and assisted by Ms. Dianne Black, Dr. Wayne Davis, and Mr.  John Sewall.




Organic chemical sample preparation and analyses were conducted under the




supervision of Drs. Lake and Rogerson, and assisted by Mr. Curt Norwood,




Ms. Sharon Pavignano, Mr. Robert Bowen, Ms.  Adria Elskus, and Mr. Lawrence




LeBlanc.  Inorganic chemical preparation and analyses were conducted under the




supervision of Dr. Gerald Hoffman, and assisted by Mr. Frank Osterman,




Mr. Warren Boothman, and Mr. Dennis Migneault.  Data management and data analy-




sis were conducted by Mr. Jerfrey Rosen and Dr. James Heltshe, respectively.




      The EPA Technical Director for the FVP was Dr. John H. Gentile; the

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Technical Coordinator was Mr. Walter Galloway, and the Project Manager was

Mr. Allan Beck.

      The study was conducted under the direct WES supervision of

Dr. Richard K. Peddicord and Dr. Thomas Dillon and under the general super-

vision of Dr. C. Richard Lee, Chief, Contaminant Mobility and Criteria Group;

Mr. Donald L. Robey, Chief, Ecosystem Research and Simulation Division;

Dr. John Harrison, Chief, EL.  The EEDP Coordinator was Mr. Robert L.  Lazor.

The EEDP Manager was Mr. Charles C. Calhoun.

      Commanders and Directors of WES during preparation of the report were

COL Tilford C. Creel, CE, and COL Robert C. Lee, CE.   Technical Director was

Mr. F. R. Brown.


      This report should be cited as follows:

      Lake, J., Hoffman, G., and Schimmel, S.  1985.   "Bioaccumulation
      of Contaminants From Black Rock Harbor Dredged Material by Mussels
      and Polychaetes," Technical Report D-85-2, prepared by the US
      Environmental Protection Agency, Environmental Research Laboratory,
      Narragansett, R. 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	    7

PART I:  INTRODUCTION	    9

      Background  	    9
      Purpose and Scope	«    9

PART II: MATERIALS AND METHODS	    12

      Sediment Collection and Preservation   	    12
      Test Species	    15
      Mussel Bioaccumulation Study  	  	    16
      Worm Bioaccumulation Study  	    23
      Chemical Analysis 	    26

PART III: RESULTS AND DISCUSSION	    38

      Mussel Test	    38
      Worm Test   	109

PART IV:  SUMMARY	138

      Mussel Bioaccumulation Study  	  	  •  138
      Worm Bioaccumulation Study  	  142
      Bioaccumulation Mussels and Worms	  143

PART V:   RECOMMENDATIONS	  .  145

      Mussels	•  145
      Worms	145
      General 	 .....  	  146

REFERENCES	«	•  147

APPENDIX A: ORGANIC CHEMISTRY DATA *

APPENDIX B: INORGANIC CHEMISTRY DATA *
* Appendices A and B were reproduced on microfiche;  they are
  enclosed in an envelope attached inside the back cover of this  report-

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                             LIST OF FIGURES
                                                                       PAGE
Figure 1.   Central Long Island Sound disposal site and south
            reference Site (41°7.95' N and 72° 52.7'West)	13

Figure 2.   Black Rock Harbor, Connecticut (73°13'W and 41° 90'N),
            source of dredged material	14

Figure 3.   Sediment dosing system with chilled water bath and argon
            gas supply	17

Figure 4.   Suspended sediment feedback control loop and strip chart
            recorder	19

Figure 5.   Blue mussel (Mytilus edulis) contaminant uptake system. * . 20

Figure 6.   Capillary column electron capture gas chromatograms
            of PF-50 (PCB) fractions from day 8 exposure,
            a. unfiltered water, b. filtrate, c. filter, d. water
            through continuous flow centrifuge (14,000 rpm) 	 45

Figure 7.   Capillary column electron capture gas chromatograms
            of PF-50 (PCB) fractions from day 8 exposure,
            a. filtrate, b.  water through continuous flow centrifuge
            (14,000 rpm)	47

Figure 8.   Capillary column electron capture gas chromatogram
            of PF-50 (PCB) fraction from mussels, time 0	49

Figure 9.   Capillary column electron capture gas chromatograms
            of PF-50 (PCB) fractions from mussels, a. time 0,
            b.  day 28 exposure	 52

Figure 10.  Capillary column electron capture gas chromatograms
            of PF-50 (PCB) fractions from exposure, a. unfiltered
            water, day 8, b. mussel, day 28	53

Figure 11.  Capillary column electron capture gas chromatograms
            of PF-50 (PCB) fraction from exposure mussels, a. day
            28, b. day 70	54

Figure 12.  Concentration of total PCBs (as A-1254) in mussels
            exposed to BRH sediment versus time. . . •	58

Figure 13.  Capillary column electron capture gas chromatograms
            of PF-50 (PCB) fraction from exposure, a. filter, day
            8, b. filtrate, day 8, and c. mussels, day 28	62

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                           FIGURES (continued)

                                                                       PAGE

Figure 14.  Capillary column electron capture gas chromatograms
            of PF-50 (PCB) fraction from exposure, a. filter, day
            8, and b. mussels, day 28	65

Figure 15.  Capillary column electron capture gas chromatograms
            of PF-50 (PCB) fractions from mussels, a. time
            0, b. control, day 28, and c. control, day 70	66

Figure 16.  EICPs from GC/MS analysis of exposure tank
            a. unfiltered water, b. filter, c. filtrate,
            d. water through continuous flow centrifuge	71

Figure 17.  EICPs from GC/MS analysis of a. BRH sediments,
            b. mussels, day 0, c. exposure tank mussels,
            day 1.8, d. exposure tank mussels, day 3.5	76

Figure 18.  EICPs from GC/MS analysis of exposure tank mussels
            from a. day 7, b. day 14, c. day 21, d. day
            28	77

Figure 19.  EICPs from GC/MS analysis of exposure tank mussels
            from a. day 35, b. day 40, c. day 49, d. day
            56	78

Figure 20.  EICPs from GC/MS analysis of exposure tank mussels
            from a. day 63, b. day 70	79

Figure 21.  Concentration of sum of parent PAHs in mussels
            exposed to BRH sediment versus time	85

Figure 22.  Capillary column  flame ionization detector gas
            chromatograms of  PF-50 fraction from a. exposure
            tank water, day 0, b. BRH sediment, and
            c. mussels, exposure day 28	87

Figure 23.  Concentration of  total petroleum hydrocarbons in
            mussels exposed to BRH sediment versus time	89

Figure 24.  Uptake and depuration of Fe, and Cr in mussels
            exposed to BRH sediment	101

Figure 25.  Uptake and depuration of Pb, and Cd in mussels
            exposed to BRH sediment	102

Figure 26.  Uptake and depuration of Cu, and As in mussels
            exposed to BRH sediment	103

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                              FIGURES  (continued)
                                                                       PAGE
Figure 27.  Uptake and depuration of Zn, and Mn in mussels
            exposed to BRH sediment	104

Figure 28.  Capillary column electron capture gas chromatograms
            of PF-50 (PCB) fraction for BRH sediment, and
            reference sediment	108

Figure 29.  Capillary column electron capture gas chromatograms
            of PF-50 (PCB) fraction for worms exposed to
            BRH sediment a. day 0, b. day  14, and c. day
            28	Ill

Figure 30.  Capillary column electron capture gas chromato-
            grams of PF-50 (PCB) fraction  for worms exposed
            to BRH sediment a. day 42, and b. day 56	112

Figure 31.  Concentration of total PCBs (as A-1254) in worms
            exposed to BRH sediment versus time	113

Figure 32.  Concentration of sum of parent PAHs in worms
            exposed to BRH sediment versus time	119

Figure 33.  Capillary column flame ionization detector
            gas chromatogram of PF-50 fraction from
            worms exposed to BRH sediment  for
            28 days    	122

Figure 34.  Concentration of total petroleum hydrocarbons
            in worms exposed to sediment verus time	123

Figure 35.  Uptake and depuration of Fe, and Cr in worms
            exposed to BRH sediment	133

Figure 36.  Uptake and depuration of Cu, Zn and Cd in
            worms exposed to BRH sediment	135

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                              LIST OF TABLES
                                                                        PAGE
1.  Summary of Experimental Conditions for Bioaccumulation
    Study with Mytilus edulis ...................... 24

2.  Mussel Dosing System; PCB Levels in Unfiltered Water ........ 39

3.  PCB Blank Levels - Mussel Dosing System Samples ........... 40

4.  Mussel Dosing System; PCB Levels in Water -Expo sure Tank ....... 42

5.  Tentative Identifications of Compounds in BCD Gas
    Chromatograms ............................ 43

6.  Comparison of Experimental and Estimated Kp's ............ 48

7.  Average Trace Metal Concentrations for Mussels Collected from
    the Exposure Chamber on Day 28 ...................  56

8.  Estimated and Measured Bioconcentration Factors (BCF) in Mussels
    at Day 28 ....................... • ...... 59

9.  Measured Log BAF for Each Separate PCB Peak in Exposed Mussels.   .  . 61

10. PAH and Ethylan Concentrations in Unfiltered Water Samples
    (in Parts per Trillion) ....................... 70

11. PAH Compounds in Mussels ...................... 72

12. Comparison of Experimental and Estimated Sediment /Water
    Partition Coefficients (Kp's) ..............  .....  75

13. Estimated and Measured Bioconcentration Factors (BCF) in Mussels
    at Day 28 .............................   80

14. PAH and Ethylan Concentrations in Exposed Mussels Expressed as
    ng/g (dry) ......... ....................  82

15. Mussel Bioaccumulation Factors (Calculated for Day 28) ....... 83

16. Levels of PAH and Ethylan Compounds in Control Mussels During
    Study ................................ 84

17. Average Trace Metal Concentrations for Black Rock Harbor
    Sediment Samples ..........................  9°
18. Seawater Metal Concentrations Determined for the Black Rock
    Harbor Sediment Exposure and Control Chambers ............  91

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                            TABLES (continued)

                                                                      PAGE

19. Average Fe/Metal Ratios for the Exposure Chamber
    Seawater and Black Rock Harbor Sediments	  .  .  93

20. Metal Bioaccumulation Factors for Mussels Exposed
    to Black Rock Harbor Sediments	  98

21.  Average Fe/Metal Ratios for Control Mussels and
     Black Rock Harbor Sediments	99

22.  PCB Bioaccumulation Factors, Exposed Worms Day 28	   116

23.  PCB Bioaccumulation Factors of Worms in Reference
     Sediment-Day 28	   117

24.  PAH and Ethylan Bioaccumulation Factors - Worms	120

25.  Mussel Bioaccumulation Factors Calculated from Filters.   .  .  .   125

26.  Comparison of BAFs from Mussels and Worms	127

27.  Average Trace Metal Concentration for Worms Collected
     from the Exposure Chamber on Day 28	131

28.  Metal Bioaccumulation Factors for Worms Exposed to
     Black Rock Harbor Sediment	136
                                         8

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                       BIOACCUMULATION OF CONTAMINANTS
                   FROM BLACK ROCK HARBOR DREDGED MATERIAL
                          BY MUSSELS AND POLYCHAETES

                            PART I:  INTRODUCTION

                                  Background

      1.  The U.S. Army Corps of Engineers (CE) and the U.S. Environmental

Protection Agency (EPA) are jointly conducting a comprehensive Field

Verification Program (FVP) to evaluate the potential environmental impact

associated with various disposal options for dredged material.  The

approach being used in the FVP is to evaluate and field validate assessment

methodologies 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 are 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 will be conducted by the EPA Environmental

Research Laboratory (ERLN), Narragansett, R.I.

                            Purpose and Scope

      2.  The aquatic disposal option of the FVP is to be used as a site-

specific case study for evaluating 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 result from the

ocean disposal of a particular material.  To measure hazard, information

on the duration and intensity of exposure (exposure assessment) of organisms

to concentrations of materials disposed at the site (predicted environmental

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concentration) is coupled with concentrations of the material determined




from laboratory toxicity studies (effects assessment) on individual




species, populations, and communities.  When properly synthesized, these




data provide an estimate of the probability of unacceptable adverse




impact on the aquatic environment as a result of the disposal of the




material.  The verification of hazard assessment is comprised of two




components: (a) documentation and comparison of the accuracy and precision




of an individual method or protocol in the lab and field, and (b) verifi-




cation of the prediction of potential impact to the aquatic environment.




Within this context, hazard assessment contains parallel predictive




laboratory and field verification components.  The achievement of the goal




of hazard assessment requires the development and verification of assessment




protocols for defining exposure and effects.




      3.  The second research component in the aquatic portion of the FVP




is an assessment of the bioaccumulation potential of available contaminants




within the dredged material by the blue mussel (Mytilus edulis) and the




polychaete worm Nereis virens.  The focus of this study is twofold!




(a) determine the qualitative and quantitative aspects of the bioavailable




contaminants within BRH dredged material which are accumulated by the




mussel and the worm; and (b) examine the uptake and depuration kinetics




of the major contaminants within the material that constitute a potential




threat to man and the ecosystem.  Results of this study will contribute




to the overall FVP by providing a predictive tool for predicting residues




of key contaminants in the fauna at the disposal site.  The accuracy




of these predictive tools will be verified in the field and reported in




a future report.
                                      10

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      4.  Chemicals of major environmental concern have three basic




characteristics: (a) they may be acutely or chronically toxic at low



concentrations; (b) they may bioaccumulate to concentrations in tissues




that cause adverse effects in the species contaminated with the




chemical, or otherwise make the species unsuitable for human consumption;



and (c) they may depurate slowly, causing a prolonged (chronic) adverse




effect or render the resource unsuitable for prolonged periods.  The




latter two concerns are addressed in this report.  The study of uptake




and depuration rates of the major bioavailable compounds and elements by




the organisms allows predictions to be made of the rate and extent of




chemical uptake and the time needed to depurate accumulated compounds




to an acceptable concentration.




      5.  Appendices A and B contain organic and inorganic chemistry




data, respectively.  Because of the extent of the accumulated data, they



were reproduced on microfiche and are enclosed in an envelope attached




to the inside back cover of this report.
                                    11

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                     PART II:  MATERIALS AND METHODS

                   Sediment Collection and Preservation

             fc
Reference Sediment

      6.  Reference sediment (REF) for the FVP 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 southernmost perimeter of the central Long Island

Sound disposal site (Figure 1).  Reference sediment was collected with a

Smith-MacIntyre grab sampler (0.1 m^) in both August and December 1982.

Sediment collected on each date was returned to the laboratory, press

sieved (wet) within 48 hr through a 2-mm mesh stainless steel screen,

homogenized, and stored at 4°C until used for experimental purposes.

Sediment was re-homogenized prior to use.

Black Rock Harbor Sediment

      7. The source of the dredged material for the FVP was Black Rock

Harbor (BRH), located in Bridgeport, Connecticut (Figure 2), with

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 bottom sediments were collected at 25 locations

within the study area using a 0.1-m2 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 chemistry analysis.  Twelve barrels were

kept at WES and thirteen barrels were transported to ERLN in a

refrigerated truck and stored at 4°C.  Prior to use the contents of each
                                    12

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                                NEW
                               HAVEN
                BRIDGEPORT
          BLACK ROCK
            HARBOR
                   LONG   ISLAND  :
                                                            FVP
                                                         DISPOSAL
                                                           SITE
                                                    SOUTH REFERENCE
                                                         • SITE
Figure 1.  Central Long Island Sound disposal site and south reference
                site (41°7.95"N and 72°52.7"W)

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     BRIDGEPORT
     N
      N
                               MAINTENANCE
                               DREDGING

                               FVP STUDY
                               REACH
       BLACK
       ROCK
       HARBOR
\\
                        400m
Figure 2.  Black Rock Harbor, Connecticut (73°13''W and 4l°90"N),
               source of dredged material
                         14

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barrel were completely homogenized and wet seived through a 1-mm mesh




seive to remove large particles.  Sediment was stored in glass bottles



at 4°C.  To verify that the contents in the bottles were consistent,




400-ml samples were taken before the 1st, 25th, and 50th bottles for




moisture content and chemical analysis.




                               Test Species




      8.  Two species of marine invertebrates were used to conduct two



separate bioaccumulation studies, including depuration phases.  A bivalve




mollusc, the blue mussel, Mytilus edulis, and the polychaete worm,




Nereis virens, were used in this study.



Mytilus edulis




      9.  The blue mussel is a filter-feeding bivalve mollusc that




ranges along the northern Atlantic coast of the United States and




Europe.  In the United States, it ranges from Maine to North Carolina




and on the Pacific coast from Alaska to California (Bayne 1976).  Mytilus




edulis was selected for this study because it is a filter-feeding mollusc,




capturing food as suspended particulates.  Species of Mytilus have been used




extensively as a biological monitor worldwide (Farrington et al. 1983)



and its biology has been studied extensively.




     10.  One month prior to exposure, adult mussels were collected




from a well-characterized area of Narragansett Bay, Rhode Island, with




relatively low background concentrations of contaminants in the sediments




(Phelps et al. 1983; Phelps and Galloway 1980).  Test organisms, 50 to 70 mm




shell length, were temperature acclimated from 5° to 10°C at the rate of




1°C per day, then held in unfiltered flowing seawater (28 to 30 °/oo




salinity) until initiation of the experiment.
                                    15

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Nereis virens




      11.  Nereis virens is a marine polychaete worm that inhabits the




coastal United States from the Gulf of St. Lawrence to the Gulf of Mexico




on the east coast and the central California coast on the Pacific Ocean




(Pettibone 1963).  They are raptorial and deposit feeders but generally




opportunistic in their feeding habits.  This species was selected because




of its deposit-feeding habits (it will feed directly on sediment



constituents), its relatively large size, and its availability.




Approximately 600 adult worms were purchased from a bait dealer in Wiscasset,




Maine, packed in wet seaweed, and shipped to ERLN.  Upon arrival at the




laboratory, they were immediately placed in sediment for testing.






                        Mussel Bioaccumulation Study




Sediment Dosing System




      12.  A sediment dosing system was constructed to provide BRH as sus-



pended sediment for the mussel bioaccumulation study (Figure 3).  The




dosing system consisted of a conical-shaped slurry reservoir placed in a




chilled fiberglass chamber, a diaphragm pump, a 4-L separatory funnel,



and several return loops that directed the particulate slurry through a



dosing valve.  The slurry reservoir (40 cm diameter x 55 cm high) contained




40-L of slurry comprised of 37.7 L of filtered seawater and 2.3 L of




BRH material.  The slurry was changed every 2-3 days during exposure.




The fiberglass chamber (94 cm x 61 cm x 79 cm high), was maintained




between 4° and 10°C using an externally chilled water source.  (The slurry



was chilled to minimize microbial degradation during the test.)  A polypro-




pylene pipe (3.8 cm diameter) placed at the bottom of the reservoir
                                    16

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       ARGON
     INJECTION
  SEPARATORY
    FUNNEL
                              DELIVERY
                              MANIFOLD
     CHILLED
     WATER  BATH
                                          DOSING
                                          VALVE
                                     TO EXPOSURE
                                       SYSTEM
                                RETURN
                                MANIFOLD
                                 SLURRY
                                 RESERVOIR
Figure 3. Sediment dosing system with chilled water bath and argon
                        gas supply
                          17

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cone was connected to the diaphragm pump  (16-  to  40-L/mln  capacity)




that had a Teflon® diaphragm.  This pump  was used to circulate  the




slurry with minimal 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 at 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




3 and 4) and back to the reservoir.   At the dosing valves, the  slurry




was mixed with Narragansett Bay  seawater which had been filtered (to 15 y)




through sand filters.   The valves were controlled by a microprocessor that




was connected to a transmissometer (Figure 4).  Under transmissometer




control, the microprocessor responds  by modulating the pulse  length to




achieve the desired setpoint of  suspended sediment measured as  turbidity




(Sinnett and Davis 1983).




Mussel Exposure System




      13.  The system used to expose  blue mussels to BRH material in the




bioaccumulation test is shown in Figure 5.  The exposure apparatus




consisted of a fiberglass, resin-coated plywood tank (123-L capacity)




partitioned into two components.  Filtered seawater entered the mixing




chamber at 2 L/min where it was  vigorously combined with the  BRH material




and marine algae as a food source (a mixture of Phaeodactylum tricornutum




and T-Isochrysis galbana).  The mixture cascaded over a partition into
                                    18

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 STRIP CHART
 RECORDER
RETURN TO
RESERVOIR
MICRO-
PROCESSOR
                          CONTROL

                            BOX,
                             I
               SLURRY
                       sssssssa
                                DOSING VALVE
                                            SOLENOID
                     EXPOSURE SYSTEM
                                 \
                                TRANSMISSOMETER

  Figure 4.  Suspended sediment feedback control loop and strip chart
                         recorder
                          19

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                                 SEAWATER/SEDIMENT
                                       SLURRY
                                            ALGAE
TO MICROPROCESSOR
 TRANSMISSOMETER
                                                    MIXING
                                                    CHAMBER
RECIRCULATING
PUMP
                     I
     Figure 5.  Blue mussel (Mytilus edulis) contaminant uptake system
                              20

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the exposure chamber containing the mussels and a transmissometer 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 transmissometer




and returned to the mixing chamber at a rate of 38 L/min.  Polypropylene




or polyethylene plumbing materials were used throughout.




      14.  The sediment dosing system delivered BRH sediment directly into




the mussel exposure chamber via the dosing valve which was controlled by




the microprocessor and transmissometer.  As the mussels removed the suspended




particles to a level below the desired concentration, the microprocessor




simultaneously opened the dosing valve to deliver the BRH suspension and




turned on a peristaltic pump to deliver algae to 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, consisting of 9 mg/L




sediment and 0.5 mg/L algae (30 million cells/L).  This concentration




of suspended sediments was estimated to be below the concentration




that would stress or adversely affect the organisms during the test




because a preliminary test demonstrated no appreciable mortality, histo-




pathological 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 ensure that




contaminants observed in the mussels were accumulated from BRH material




rather than from the seawater or the algal cultures.  The control exposure
                                    21

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was conducted in an identical test apparatus, but no sediment was




delivered to the chamber.  Instead, a suspended particulate concentration




of 0.5 mg/L consisting entirely of algae was maintained by the micro-




processor feedback system.




Experimental Conditions




      16. 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) 1982).  Although not specifically




intended for suspended sediment testing, the general recommendations




defining test animal care, handling, acclimation procedures, seawater




quality, and acceptable exposure conditions were suitable for this test.




      17.  At the start of the bioaccumulation study, 300 mussels




were initially placed in each of the BRH and control chambers.  Before




placing the animals in the test chamber, 20 animals were randomly




selected for organic and inorganic chemical analysis to determine the




baseline residues in the mussels before the exposures began.  During




the test, 20 mussels were sampled for chemical analysis on days 1.8,




3.5, 7, 14, 21, and 28 during exposure and 35, 40, 49, 56, 63, and 70




during depuration in the BRH chamber, and 20 mussels were sampled on




days 28, 56, and 70 in the control chamber.  To avoid excessive loading




of the tanks, the shorter exposures were conducted after some of the




mussels had been removed by sampling.  Specifically, the mussels for days




1.8 and 3.5 exposures were placed in the tank on day 14 and removed on




days 16 and 18 respectively.  Likewise, mussels for the 7 day exposure




were placed in the tank on day 21 and removed on day 28.
                                    22

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Since this design assumes that the exposure system operates in a




consistent manner, two 14-day exposures were conducted to verify con-




sistency, one from day 0 to day 14 and a second from day 14 to day 28.




      18.  Twice each week suspended particulate concentrations from the




control exposure chambers were analyzed by dry weight determination and




by electronic particle counting (1 to 40 u particle range).  The dry




weight determinations were conducted according to Standard Methods




(American Public Health Association (APHA) 1976) with the following




modifications.  Before sample filtration, filters were washed with a 50-




ml aliquot of delonized water, then with three 10-ml aliquots of deionized




water.  Following filtration, filters were rinsed with three 10-ml rinses




of 2.4 percent ammonium formate to remove salt.  Measurements of dissolved




oxygen, salinity, temperature, and ammonia nitrogen were made to determine




water quality and are presented in Table 1.




                        Worm Bioaccumulation Study






      19.  The worm bioaccumulation study consisted of an exposure of




Nereis^virens to solid phase BRH or REF materials for as long as 40




days under flowing seawater conditions.  Twenty-four hours prior to




introducing the animals to the exposure aquaria, approximately 9.5 L




of either sediment was placed in aquaria measuring 32 cm x 38 cm x 16




cm high.  Ambient temperature (9° to 13°C) seawater was then provided




to each of 14 aquaria at the rate of 120 ml/min.  Sediment depth in




each aquarium was approximately 8 cm; seawater depth (maintained by a




standpipe) was approximately 5 cm.




      20.  The test was initiated at time zero (TQ) by randomly placing




24 adult worms in each of 12 aquaria.  Nine aquaria contained BRH
                                    23

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                                   Table  1

            Summary of  Experimental Conditions  for Bioaccumulatlon
                         Study with Mytilus  edulis*
    Parameter
  Control
  Exposure
Suspended solids
   dry wt, mg/L

Particle density,
   No./L

Temperature, °C
Dissolved oxygen,
   mg/L

Salinity, °/oo
Unionized ammonia,
Uptake Period

 1.72 + 0.18
(1.45 - 2.02)

 2.60 + 0.2 X 107
(2.00 - 3.1 X 10?

 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)
 9.32 + 0.58
(8.19 - 10.33)

12.00 + 1.3 X 10?
(9.60 - 13.7 X 10?)

 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)
Suspended solids
   dry wt, mg/L

Particle density,
   No./L

Temperature, °C
Dissolved oxygen,
   mg/L

Salinity, c/oo
Unionized ammonia,
   ug/L
Depuration Period

 23.5 + 1.17
(1.18 - 3.53)

  2.8 + 0.3 X 10?
 (2.5 - 3.2 X 10?)

 15.2 + 0.3
(15.0 - 15.8)

  8.0 + 0.2
 (7.6 - 8.5)

 27.9 + 1.9
  (23 - 30)

 1.30 + 0.26
(0.94 - 1.66)
 2.48 + 1.64
(1.16 - 4.86)

  2.9 + 0.2 X 107
 (2.7 - 3.2 X 107)

 15.2 + 0.3
(15.0 - 15.8)

  8.0 + 0.3
 (7.6 - 8.4)

 27.9 + 1.9
  (23 - 30)

 1.33 + 0.22
(0.96 - 1.52)
* Tabular values are mean and standard deviation with range denoted in
  parentheses.
                                     24

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material and three REF sediment.  A subset of 15 worms was randomly



selected for organic and inorganic chemical analysis to determine the



contaminants in worms at T0.  Prior to chemical analysis, all worms




at all sampling times were placed in Petri dishes containing filtered




seawater and allowed to purge their gut contents for 14 hr.



      21.  For each sampling period, all the worms from a single aquarium




were removed for analysis.  Sampling periods during the uptake portion




of the study were days 14 and 28.  After 28 days, all worms in four of




the five remaining BRH aquaria were removed, the sediment emptied, and




the aquaria cleaned.  The aquaria were then filled with REF sediment



and the worms placed back into the aquaria.  Sampling of these worms




during the depuration phase was on days 42 and 56 (14 and 28 days of




depuration).  The worms in the ninth BRH aquarium were allowed to




remain an additional 12 days (total of 40 days exposure) and archived



at -20°C.




      22.  Three aquaria were each provided with REF sediment and




24 worms.  The worms were sampled on days 28, 40, and 56 to determine



what contaminants, if any, were obtained from the REF sediment.



     23.   For clarity, mussels exposed to BRH sediment and algae are




referred to as "exposed mussels," while mussels exposed to algae only




are referred to as "control mussels,"  For the worn study, worms exposed




to BRH sediment are "exposed worms," while those depurated in reference



sediment are referred to as "depurated worms."  Worms exposed to reference




sediments are referred to as "reference worms."
                                    25

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                              Chemical Analysis






Organic Sample Preparation




      24.  The analytical procedures described below represent  the state




of the art in marine organic analysis and have been intercalibrated with




several oceanographic laboratories.  EPA-recognized analytical  methods,




while available for these classes of contaminants, have been developed




primarily for freshwater and wastewater systems.  These methods require




extensive modification and intercalibration when applied to marine systems




for the types of matrices and levels of detection that are required in this




study.




      25.  Cleaning of Glassware and Equipment.  All glassware  used for




the collection, storage, extraction and analysis of samples was washed




with Alconox®, rinsed four times with hot tap water, four times with




deionized water, capped with aluminum foil, and muffled for 6 hr at 450°C.




Immediately prior to use glassware was rinsed three times with  an appropriate




solvent.




      26.  Stainless steel centrifuge bottles were washed in the same




manner as glassware and then rinsed twice with methanol, twice with




methylene chloride and twice with hexane immediately prior to use.




      27.  Stainless steel tissue homogenizers were washed in the




same manner as glassware and then placed in an ultrasonic bath  in




graduated cylinders filled first with methanol, then with methylene




chloride, and finally with hexane just prior to use.




      28.  Glass fiber filters were placed individually in aluminum foil




and muffled for 6 hr 450°C.  The stainless steel filter housing was




washed and rinsed with acetone and hexane prior to use.
                                    26

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      29.  Sediment.  The methods that follow were used for the extraction




and analysis of BRH sediment and the reference sediment from the worm




dosing system.  Approximately 10 gr of wet sediment was placed in a




stainless steel centrifuge tube, and 50 ml of acetone was added.  The




mixture was homogenized for 40 sec using a brass-bearing-equipped




tissue homogenizer and then centrifuged at 10,000 RFM for 5 min.  The



acetone was decanted into a 1-L separatory funnel containing 150 ml of




pre-extracted deionized water.  The extraction and centrifugation steps




were repeated twice 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 combined in




a 500-ml Erlenmeyer flask.  Extracts were frozen to remove water.  The




sample extract was then subjected to column chromatography (see Column




Chromatography, paragraphs 39 and 40).




      30.  Water.  The following procedure was used for unfiltered water



samples (dissolved plus particle-bound contaminants), samples of filtered




water collected after the glass fiber filter, and water taken after




passage through a continuous flow centrifuge.  Water samples were collected



in 6-L separatory funnels.  Samples were extracted twice by the addition




of 100 ml Freon 113 followed by vigorous shaking.  Extracts were combined




in a 500-ml Erlenmeyer flask, and sodium sulfate (previously muffled at




700°C for 4 hr) was added to remove water.




      31.  The Freon extract was poured off and volume reduced in a round




bottom flask fitted with a Kuderna-Danish evaporator, and the solvent was




changed to hexane.  Extracts (5 ml) were fractionated using the second silicic




acid column (see Column Chromatography, paragraphs 39 and 40).
                                    27

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       32.  Suspended Particulate Material.  For the mussel study suspended




particulate material (SPM) was collected using a 273-mm glass fiber filter




(Gelman Type'AE, 0.1 micron) in a stainless steel housing (Millipore®




273 mm).  Water from the exposure system tanks was allowed to gravity




feed into this filtering system through Teflon® tubing.




      33.  Each filter was carefully removed, placed in a stainless




steel centrifuge bottle, and frozen until preparation and analysis.




Acetone (50 ml) was added to the centrifuge bottles containing the




filter, and the filter was homogenized with a stainless steel tissue




homogenizer for 20 sec at 25,000 RIM.  Samples were centrifuged at 10,000




RPM for 5 min, and the acetone water layer was decanted into a 1-L




separatory funnel containing 150 ml extracted deionized water.  This




extraction procedure was repeated two more times using 50 ml of Freon




113.  The Freon was added to the separatory funnel, which was then




shaken.  The Freon layer was then drawn off and saved.  The remaining




aqueous layer was extracted again with 50 ml of Freon, and the extracts




were combined.  The sample extract was then subjected to column




chromatography (see Column Chromatography, paragraphs 39 and 40).




      34.  Organisms.  Mussel samples were taken for background analysis




at day zero, and removed from the exposure tank at day 1.8, 3.5, 7,




14, 28, 35, 40, 49, 56, 63 and 70; control mussels were sampled on days




28, 56, and 70.  At each sampling time, 20 mussels were removed using a




stratified random sampling plan and stored in muffled aluminum foil in a




freezer prior to analysis.  From each group of 20 mussels, three replicates




consisting of four individuals each were shucked into pre-weighed glass




centrifuge tubes, homogenized with a tissue homogenizer for 20 sec, and
                                    28

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centrifuged at 25,000 RPM for 5 min.  The remaining 8 organisms were




archived at -20°C.



      35.  Dead mussels were removed daily when discovered and mortality




recorded.  Mortality data were analyzed by calculating survivorship




functions for mussels from control and exposed treatment conditions.




These survivorship functions incorporated the effect of the periodic



removal of individuals for analyses other than mortality.  A comparison




of these functions was made using the Mantel and Haenszel (1959) Chi




square test for comparing two survival distributions.



      36.  Worms were removed from exposure tanks for chemical analysis



on days 14, 28, 42, and 56 and from the control tanks on day 28.  A




sample was also collected at day 0, prior to exposure.  Following




collection from the experimental tanks and gut depuration (see Methods




paragraph 20), worms were frozen until analysis in muffled glass jars.




From these samples, three replicates of 1-2 individuals each were placed




into preweighed glass centrifuge tubes, homogenized with a tissue




homogenizer for 20 sec and centrifuged at 25,000 RPM for 5 min.




      37.  Approximately 2 g of the mussel and worm homogenates was




taken for inorganic analysis.  A small portion (approximately  2 g) was




taken for wet:dry ratio determinations.  The remaining homogenate was




weighed and used for organic analysis.




      38.  Each of the 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; the mixture was




then homogenized with a tissue homogenizer for 20  sec and centrifuged




at  1750 RPM for 5 min.  The fluid  layer was decanted into a separatory
                                     29

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funnel containing  150 ml of pre-extracted deionized water.  The acetone




extractions and centrlfugation were repeated once more and the extracts




were combined in the separatory funnel.  The tissue homogenization,




extraction, and centrlfugation were repeated twice more using 25 ml of




Freon 113 as the solvent.  Because of the density of the 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 was drawn off and saved.  The




remaining aqueous  layer was extracted twice more with 50 ml of Freon




each time.  The Freon extracts were combined and the aqueous layer was



discarded.  The sample extract was then subjected to column chromatography




(see Column Chromatography, paragraphs 39 and 40).




      39.  Column  Chromatography, Final Volume and Storage.  To remove



interfering biogenlc material and some residual particulates, the combined




Freon extracts were passed through the first column (2 x 25 cm of 100%




activated 100-200 mesh silicic acid).  For sediment samples, 2.5 cm of




activated copper powder was added to the bottom 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 evaporator




and 3-ball Snyder column.  The solvent was exchanged to hexane as the




sample approached  5 ml.  Final volume reduction to 5 ml was accomplished




by placing the sample in a concentrator tube fitted with a mlcrosnyder column




and placing it into a tube heater.




      40.  The 5-ml sample extracts were then charged onto a 0.9 x 45 cm




second column of 5% water deactivated 100-200 mesh silicic acid.  Three
                                    30

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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 is an expansion of a 1st fraction formally
used by this laboratory.  The PF-50 fraction is designed to include PCBs
and related chlorinated pesticides of similar polarity in addition to a
large portion of the petroleum hydrocarbons.  Petroleum hydrocarbons
(PHC) as referenced in this report include only those hydrocarbon compounds
in the PF-50 fraction.  The polycyclic aromatic hydrocarbons (PAH) which
are collected in the F-2 fraction may also be of petroleum origin; however,
these PAH compounds and the small amount of unresolved material found in
the F-2 (as separated in the present study) represented only a small portion
(approximately 10%) of the total petroleum hydrocarbons and were not
included in petroleum hydrocarbon calculations.   The F-3 fraction
collected more polar material.  Each column fraction was reduced in volume
by a Kuderna-Danish evaporation 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,
microsnyder columns, and a tube heater were utilized to reduce the sample
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 GC/MS analyses.
Organic  Instrumental Analysis
       41.   Electron capture  gas chromatographic analyses were conducted on
a Hewlett-Packard Model 5840 gas chromatograph equipped with a 30 meter DB-
 5 fused  silica capillary column from J & W.   The chromatograph was
                                     31

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temperature programmed from 80°C to 290°C at 10°C/mln with a 4-min




hold at 80°C.  Flame ionization gas chromatographic analyses were conducted




on a Carlo Erba 4160 gas chromatograph equipped with an identical




column.  The temperature was programmed from 60°C to 325°C at 10°C/min




with a 4-min hold at 60°C.




      42.  Gas chromatograph/mass spectrometric (GO/MS) analyses were




conducted on a Finnigan Model 4500 also equipped with J & W DB-5 30




meter fused silica capillary column.  The tail of the capillary column




was positioned inside the mass spectrometer so that the effluent




from the column was directed into the ionization volume of the mass




spectrometer.  The mass spectrometer was operated through a standard




Incos data system and was tuned at all times to meet EPA quality assurance




specifications using decafluorotriphenylphosphine.  The ionizing current




was typically set at 300 milliamperes and 70 EV, and the instrument




operated such that 100 picograms of PAHs 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 typically programmed from 50°C to 330°C at 10°C/min with a 2-min



hold at 50°C, but was occasionally progammed at 4°C/min to permit higher




chromatographic resolution.




      43.  All instruments were calibrated with standards each day.  The




concentrations of the standards 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.  When standards




were not available for some compounds, response factors were calculated




using mean responses of appropriate standards.
                                    32

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jnorganic Sample Preparation



      44.  Seawater.  Two sets of seawater samples were collected from



the mussel exposure system on day 25 of the exposure period.  Approximately




1 hr before the BRH sediment slurry was renewed in the reservoir, duplicate




20-ml samples of seawater were taken from the control and exposure chambers.



Two 20-ml seawater samples were also taken 3 hr after renewing the slurry




from the exposure chamber only.  The unfiltered samples were acidified




with 2.0 ml of ultra-pure concentrated nitric acid and placed in acid-




cleaned polyethylene bottles fitted with polyethylene screw caps.  The



acidified samples were stored at room temperature for 1 week before




trace metal analysis.




      45.  Sediment.  After the BRH sediment contained in a barrel




was thoroughly homogenized (see Sediment Collection and Preservation,




paragraphs 6 and 7), nine samples were taken for analysis.  These samples



included three from the top, three from the middle, and three from the




bottom.  The wet weight of all samples was determined.  The samples




were frozen and then freeze dried in a Virtus® lypholyzer (Model #




10-145MR-BA) for 2 days.  The dry weight of each sample was then



determined.




      46.  The dried BRH sediment samples 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 for several days.




The samples were subsequently evaporated down to approximately 10 ml




after which 30% H202 was added in 2-ml aliquots until 50 ml had been
                                    33

-------
added.  The H202 was added cautiously  since  BRH  sediment  reacts vigorously



with strong oxidizing agents.  The samples were  evaporated  down to




approximately 25 ml and filtered through acid-rinsed  (5%  HNC^) Whatman




41 filter paper into 250-ml volumetric flasks.   The beakers were rinsed




with 25-ml quantities of  5% HNC>3.  The-rinse solution was also filtered




through the filter paper  and added to  the volumetric  flask.  The volumetric




flasks were brought up to volume with  5% HN03.   This  nitric acid-hydrogen




peroxide extraction procedure for sediment samples has been described by




Krishmanurty et al. (1976).




      47.  Organisms.  From each sample homogenate, described in Organic




Sample Preparation, about 2 g of wet tissue  was  taken for inorganic




analysis and 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 concentrated reagent grade nitric  acid was added to




each sample, which was then allowed to digest at room temperature in a




hood for 24 hr.  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 were  slowly added in 1-ml




increments since the effervescent reaction was quite  vigorous.  The




solutions were then heated to 60°C for another day, 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 finally transferred  to screw cap poly-
                                    34

-------
ethylene bottles.  This nitric acid-hydrogen peroxide dissolution procedure




has been reported by Knauer and Martin (1973).




Inorganic Instrumental Analysis




      48.  All flame atomization (FA) atomic absorption (AA) analysis




was conducted with a Perkin-Elmer (Model #603) atomic absorption instrument.



All Hg determinations were conducted by the method of Hatch and Ott (1968)




using a Perkin-Elmer (Model #MHS-1) mercury/hydride system adapted to




the 603 AA.  The transient Hg signals were recorded with a Perkin-Elmer



(Model #56) strip chart recorder.  All heated graphite atomization (HGA)




atomic absorption determinations were conducted with a Perkin-Elmer



(Model #500) HGA unit coupled to a Perkin-Elmer (Model #5000) atomic




absorption instrument retrofitted with a Zeeman HGA background correction




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 setup 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.




      49.  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
                                    35

-------
flask.  All samples were analyzed at  least  twice  to determine signal




reproduclbility.  Most were analyzed  three  times.  Generally, for each




15 samples processed, one sample was  determined by the method of standard




addition, and one procedural blank sample was analyzed.




      50.  All elements except Hg and As were determined in the sediment




samples by FA-AA.  Mercury was determined only in the BRH sediment




samples by the 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.



      51.  All seawater samples were  analyzed by H6A-AA.  No chemical




separation techniques were utilized to concentrate the elements of interest




from the seawater matrix.  All samples were analyzed by direct injection




into the HGA unit (Ediger et al. 1974; Sturgeon et al. 1979; and Slavin



1980).  The large non-atomic background signal was eliminated by the use




of the Zeeman background correction system  (Fernandez et al. 1980, and




Fernandez and Giddlngs 1982).  It was necessary to matrix match the




unknown samples with the standards since chemical interferences are not




corrected by the Zeeman effect.  Therefore, all standards were prepared



in trace metal stripped seawater and  acidified in the same manner as the




samples.  The trace metal-free seawater was prepared by the methods of




Davey et al. (1970).




      52.  Due to the limited size of the mussel and worm samples (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 and worm samples determined by HGA-AA were matrix matched




before analysis.  A matrix solution containing 10% seawater and 90% 0.16
                                    36

-------
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.




      53.  It should be noted that, unlike the As determined in BRH




sediment samples, no chemical interference was detected for the As




determined in the mussel samples.  There is a large difference in the




two sample matrixes with respect to the inorganic and organic composition




which could account for the absence or presence of a chemical interference




during the determination of As by HGA-AA or MHS-1 AA analysis.
                                    37

-------
                     PART  III:   RESULTS  AND  DISCUSSION

                                Mussel Test

Organic Contaminants

       54.   PCBs - Unfiltered Seawater.  The PCB concentrations  (quantitated

as Aroclor-1254) in whole water samples (dissolved plus particle bound

PCB compounds) taken during the exposure and depuration phases  of  the

bioaccumulation study are shown in Table 2.  The  PCB  concentrations found

in blanks processed through the analytical  procedure  averaged 0.21 ng/1

(Table 3).  The average concentration of PCBs  (as A-1254) found in control

tanks was 0.52 ng/1.  PCBs found in unfiltered seawater samples from the

exposure tanks showed an average concentration of 112+29.3 ng/1 during

the exposure.  During the exposure the RSD*  of the measurement  for

total PCBs was 20%, indicating  that the exposure system was working well

and that it delivered a relatively constant  concentration of PCB

contaminants to the mussels.  During the depuration period the  PCB

concentrations in the exposure  tank decreased; however, they remained

elevated above those in the control tank during the depuration  period

(Table 2).  Since the fiberglass exposure tank was cleaned with soap and

water and thoroughly rinsed following the exposure period, the  elevated

concentrations found in the tank during depuration may reflect  the further

introduction of contaminants from a variety  of possible sources (i.e., par-

ticulates associated with the mussels1 shells  or byssal threads, feces and

pseudofeces, etc.).
*  RSD - Relative standard deviation = standard deviation  x 100
                                             mean
                                    38

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                                  Table 2




            Mussel Dosing System; PCS Levels in Unfiltered Water



Day
0
0
28
28
35
70
70



Day
0
0
8
8
14
14
16
16
21
21
*28
28

35
70
70
CONTROL TANK

Replicate
A
B
A
B
A
A
B

EXPOSURE TANK

Replicate
A
B
A
B
A
B
A
B
A
B
A
B

A
A
B

PCB (as A-1254)
ng/1 (not corrected for blank levels )
0.46 .50 + .05
0.53
0.23 .34 + .16
0.45
0.65 .65
0.66 .66 + .00
0.66
.52 + .16

PCB (as A-1254)
ng/1 (not corrected for blank levels)
95.3 115. + 27.2
134.
116. 123. + 9.2
129.
97.4 80.2 + 24.3
63.
133. 155. + 31.1
177.
80.3 91.7 + 16.1
103.
105. 107. + 2.8
109.
112. + 29.3
2.27
1.79 1.83 + 0.06
1.87
*At day 28 exposure ended and depuration period began.




                                     39

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                            Table 3


        PCS Blank Levels - Mussel Dosing System Samples*



                                              PCB (as A-1254)
       Day                                         ng/1

  24 February 83                                    .18

   2 March 83                                       .16

   4 March 83                                       .16

   9 March 83                                       .34

  16 March 83                                       .24

  23 March 83                                       .20
                                                    .21 + .07
* For PCB levels given (Table 2), the blank levels have not
  been subtracted.
                            40

-------
      55.  The distribution of PCB compounds between the dissolved**

and particle-bound form was examined in samples of the exposure water.

Both the filters and the filtrate were extracted and analyzed.  Another

separation of dissolved and particulate phases was accomplished using a

continuous flow centrifuge.  The results of these studies are shown in

Table 4.  The mean PCB concentration for the filters (day 8) added to the

mean PCB concentration for the dissolved compounds (day 8) is close to

the value obtained for analysis of unfiltered water on day 8.  These data

indicate that methylene chloride method for extracting PCB compounds from

the suspended particulate suspensions was as efficient as the sum of the

individual extractions of the filtrate and the particles (see Methods).

        56.  The electron capture detection gas chromatograms from the

analysis of unfiltered water, filters, filtrate, and centrifuged water

taken from the dosing system on day 8 of exposure are shown in Figure 6.

The chromatogram of the unfiltered water (dissolved and particle-bound

contaminants) shows a distribution of PCB compounds from Cl2 to Cl% with

the majority of material containing four, five, and six chlorine atoms.

Tentative identification of the compounds in electron capture chromatograms

are shown in Table 5.  The same general patterns of peaks are shown in

the filter sample; however, there appears to be a relative decrease in
** Dissolved as used in this report refers to the compounds passing
   through the O.!-(JL glass fiber filter and that material which passed
   through the continuous flow centrifuge.  These compounds may be
   associated with surfactants or may be in colloidal forms and not
   truly dissolved.
                                     41

-------
                                  Table  4
          Mussel Dosing System; PCS Levels in Water-Exposure  Tank
    Day

Unfiltered Water

     8
     8
Replicate
    A
    B
           PCB (as A-1254)
ng/1 (not corrected for blank levels^
     116.
     129.
123.   + 9.2
Filtered Water

     8
     8
    A
    B
      11.1
      12.0
 11.6 + .64
Water thru Centrifuge at  14,000 RPM
Filter
                                                 10.6
                                                108.
                                      42

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                                     Table  5
         Tentative  Identifications  of  Compounds  tn BCD Gas  Chromatograms*
   Peak It         Tentative  ID

      1            2,3 - Dichlorobiphenyl
      2            Dichloroblphenyl
      3            Dichlorobiphenyl
      4            2,2',5 -  trichlorobiphenyl
      5            Trichlorobiphenyl
      6            Trichlorobiphenyl
      7            Trichlorobiphenyl
      8            Trichlorobiphenyl
      9            2,4',5 -  trichlorobiphenyl
     10            2,4,4' -  trichlorobiphenyl
     11            2,3,4  -  trichlorobiphenyl
     12            Trichlorobiphenyl
     13            Trichlorobiphenyl
     14            Tetrachlorobiphenyl
     15            2,2',4',5,  - tetrachlorobiphenyl
     16            2,2',4,4' - tetrachlorobiphenyl
     17            2,2',3',5 - tetrachlorobiphenyl
     18            Tetrachlorobiphenyl
     19            Tetrachlorobiphenyl
     20            Tetrachlorobiphenyl
     21            2,3',4',5 - tetrachlorobiphenyl
     22**          2,3',4,5',6 - pentachlorobiphenyl, 2,3',4,4' - tetrachlorobiphenyl
                      2,2',3,5,6 - pentachlorobiphenyl
     23            Pentachlorobiphenyl, 2,3,8 - trichlorodibenzofuran,
                                                  tetrachlorodiphenyl ether
     24            Tetrachlorobiphenyl, Pentachlorobiphenyl
 *Since all PCB isomer standards were not available, the possibility exists that
  other isomers nay elute with identical retention times as the PCB in this table.
  Therefore we prefer the conservative approach by listing identifications as
  tentative.

**More than one PCB isomer standard with this retention time eluted in this
  position.
                                         A3

-------
                             Table 5. (Cont'd)
Peak //         Tentative ID

  25            2,2',4,5,5'- pentachlorobiphenyl
  26            Pentachlorobiphenyl
  27            Pentachlorobiphenyl
  28            Pentachlorobiphenyl, 1,1 - bis (p-chlorophenyl) - 2,2-dichloroethylene
  29            Pentachlorobiphenyl
  30            Pentachlorobiphenyl
  31            Pentachlorobiphenyl, Hexachlorobiphenyl
  32            Pentachlorobiphenyl, Hexachlorobiphenyl
  33            Pentachlorobiphenyl
  34            Pentachlorobiphenyl, Hexachlorobiphenyl
  35            Hexachlorobiphenyl
  36            2,2',4,4',5,5' - hexachlorobiphenyl
  37            Pentachlorobiphenyl, Hexachlorobiphenyl
  38            Hexachlorobiphenyl
  39            Hexachlorobiphenyl,
  40           2,2',3,3',4,5 -hexachlorobiphenyl
  41           Heptachlorobiphenyl
  42           2,2',3,4,4',5',6- heptachlorobiphenyl
  43           2,2',3,3',4,4'- hexachlorobiphenyl
  44           Hexachlorobiphenyl
  45           Heptachlorobiphenyl
  46           2,3,3•,4,4',5- hexachlorobiphenyl
  47           2,2',3,3',4,5',6,6'- octachlorobiphenyl
  48           Heptachlorobiphenyl
  49           Heptachlorobiphenyl
  50           Octachlorobiphenyl
  51           Octachlorobiphenyl
  52           2,2',3,3',4,4',5,5' -octachlorobiphenyl
  53           2,2',3,3',4,4',5,5',6 - nonachlorobiphenyl
  54           Decachlorobiphenyl
                                         44

-------
                               Time
                                                                                       Time
                      a.   Unfiltered water
                                                                                  b.   Filtrate
•is
                                                                                      Time
                                                                     d.   Water through continuous flow
                                                                           centrifuge (14,000 rpm)

                     Figure 6.  Capillary column electron  capture  gas  chromatograms of PF-50
                                         (PCB) fractions from  day  8  exposure

-------
 the height  of  the  lower  molecular weight  peaks  in comparison with the

 chromatogram of  the  unfiltered  water.   The  chromatogram of  the  filtrate  shows

 a  relative  enhancement of  the lower  molecular weight  PCB compounds when

 compared with  the  unfiltered water.  The  distributions  found are  logically

 consistent  with  the  solubilities of  the compounds.  With lower  molecular

 weight, more water-soluble PCB  compounds  were found in  the  filtrate and

 the higher  molecular weight, less soluble compounds were found  associated

 with particles.

       57.   In  order  to determine whether  the  distributions  found  on the

 filter and  in  the  filtrate were artifacts of  the  filtration process (i.e.,

 adsorption  of  less soluble PCB  components on  the  filter while more soluble

 components  passed  into the filtrate), continuous  flow centrifugation at

 14,000 RPM  was utilized  to remove particles.  Analysis  of the water

 following passage  through  the centrifuge  showed a distribution  of PCB

 compounds that was very  similar to the  distributions  found  in the filtrate

 (Figure 7).  While PCBs  in the  water passing  though the centrifuge  may

 still be associated  with extremely fine particles or  exist  in colloidal

 form, this  experiment showed that the separations were  not  artifacts of

 the filtration process.

      58.   Data  from the analysis of filtered material  and  filtrates were

utilized to calculate sediment-water partition coefficients,  Kp,  where

                         Kp = Cs_
                              Cw

              Cs» concentration of compound in sediment

              Cw» concentration of compound in water

Kps were estimated for PCB compounds where Log P  (Log of the  n-octanol/

water partition coefficient) were known (Table 6).  The estimated
                                    46

-------
    t
    c
    o
    Q.
    cr
    o
    o
                     NUMBERS ABOVE PEAKS REFER TO IDENTITIES
                     OF COMPOUNDS LISTED IN TABLE 5.
                                           Time
                                 a.  Filtrate
                                                   15
                                                         30
                                       Time
        b.   Water through  continuous flow  centrifuge  (14,000 rpm)


Figure  7.   Capillary colunm  electron capture  gas chromatograms of PF-50
                    (PCB) fractions from day 8 exposure
                                      47

-------
                                    Table 6

                 Comparison of Experimental and Estimated Kps
Peak No.    PCS Compound                     Log P*    Kp Experimental**  Kp Estlmatedt

   4     2,2,'5 - trichlorobiphenyl           4.7         .99 X 105          .02 X 105

  10     2,4,4' - trichlorobiphenyl           5.0         2.1 X 105         .036 X 105

  25     2,2',4,5,5'-pentachlorobiphenyl      6.3         12. X 105          .66 X 105

  36     2,2l,4,4',5,5'-hexachlorobiphenyl    6.7         42. X 105          1.8 X 10$

  43     2,2',3,3',4,4'-hexachlorobiphenyl    7.0         51. X 1Q5          3.2 X 105
   * Solubility from Mackay et al.(1980b).  Converted to Log
     P using Log P • 5.00-.670 Log S where S is solubility in pmol/1 (Chlou
     et al. 1977).

  ** Kp measured as means of 3 Kps determined on day 28 from mussel exposure tank,

   t Kp estimated using Log Koc - Log Row - 0.21
       Row - n-octanol/water partition coefficient (Log P)
       Koc » organic carbon/water partition coefficient from (Karickhoff  et
       al. 1979)

     and Kp - Koc (%OC) from Briggs (1973).
                   100
                                          48

-------
                  NUMBERS ABOVE PEAKS REFER TO IDENTITIES

                  OF COMPOUNDS LISTED IN TABLE 5.
                                                                                   35
t
0)


o
a.
m
OJ
a:
 o
 
-------
results were considerably lower than the measured results for representative




PCB Isomers.  This may indicate that equilibrium was not established with




respect to PCBs in the aqueous and particle bound phases during  the




residence time of the suspensions in the dosing system.




      59.  PCBs - Mussels*  An electron capture detection (BCD)  gas




chromatogram from mussels taken at day 0 (Figure 8) shows a pattern of




electron capturing compounds that is typical of mussel samples from lower




Narragansett Bay (Lake et al. 1981).  This chromatogram shows a  peak




consisting of 2,4,8-trichlorodlbenzofuran and a tetrachlordiphenyl




ether (which co-elute under the gas chromatographic conditions employed).




The predominant peaks are Clg PCBs.




      60.  Following exposure to suspensions of BRH material (day 28),




more PCB peaks are evident in the BCD chromatograms of mussels and the




distributions are changed considerably (Figure 9).  In particular, the




lower molecular weight PCB compounds consisting of PCBs with two, three,




and four chlorine atoms are significantly increased, and the maximum




peaks consist of Cl5 compounds.  Relative increases in some Cl^  and




Cly peaks eluting in the later portions of chromatograms are also




evident.




      61.  Comparison of this chromatogram with that of the unfiltered




dosing water (Figure 10) shows that the mussels accumulated most of the




different PCB isomers present in the unfiltered water.  The organisms appear-




ed to show a distribution which was very similar to that in the unfiltered




water; and,  as was observed in other studies (Lake et al. 1983), PCBs with




seven or more chlorine atoms were not accumulated as effectively as




those with four, five, and six chlorine atoms.
                                    50

-------
      62.  The chromatogram from mussels exposed for 28 days, and the




chromatogram of mussels exposed for 28 days followed by 42 days of




depuration, are shown In Figure 11.  Comparison of these chromatograms




shows relative decreases in lower molecular weight PCB compounds (Cl2,




013, and 014 isomers) and in some Gig and Cly PCB isomers in the




depurated sample, as well as relative increases in other Cl$ isomers




(peaks No. 36 and 39).  The peaks which are becoming more prominent are




the same PCB peaks that are predominant in the chromatograms from




control mussels and mussels from lower Narragansett Bay.



      63.  Since mussels were not gut purged in the present study, the




extracts of mussels include a PCB contribution from SPM in the gut of the




organisms.  While the significance of this material to the total PCB



content of the organisms has not been determined in the present study by




examining gut-purging, two facts support the contention that it is not




dominant in determining the PCB distributions in mussels.  First,




research examining the uptake of PCBs in similar dosing studies found no




differences in PCB concentrations between non gut-depurated mussels and




mussels depurated for 6 hr (Pruell et al.  1983).  In addition, these




researchers found the SPM contained C18, Clg, and C110 P088 which



were not observed in the mussel extracts but which would have been present




if material in the gut had significantly influenced the PCB contaminants



in the extracts.  Secondly, the amount of  SPM present in the organisms




at day 28 can be calculated from the accumulation of Fe  (which is not




highly bioaccumulated) and the concentration of Fe in the  BRH sediment.




The  amount of sediment accumulated multiplied by the concentration of
                                     51

-------
  t
  U>
  §
  o.

  I
              NUMBERS ABOVE PEAKS REFER TO IDENTITIES
              OF COMPOUNDS LISTED IN TABLE 5
                                                  40
                                      Time
                                   a.   Time 0
Figure  9,
                          Time	>


                 b.  Day  28  exposure



Capillary  column electron capture gas  chromatograms of  PF-50

             (PCB) fractions from mussels
                                       52

-------
   t
   O)
   

   §
   Q.
   «
   0)
   Q
                 NUMBERS ABOVE PEAKS REFER TO IDENTITIES
                 OF COMPOUNDS LISTED IN TABLE 5.
                                                                  50
                                                                 JILj^-jJuJI	
                        a.
          Time 	>


Unfiltered  water,  day
                                       Time 	>

                             b.   Mussel, day  28



Figure 10.   Capillary column  electron capture  gas chromatograms of PF-50

                       (PCB)  tractions from  exposure
                                       53

-------
            NUMBERS ABOVE PEAKS REFER TO IDENTITIES
            OF COMPOUNDS LISTED IN TABLE 5.
                                      Time	>

                                   a.  Day  28
Figure  11.
                           Time 	>

                     b.   Day 70

Capillary  column electron capture  gas chromatograms of PF-50
        (PCB) fraction from exposure mussels

-------
PCBs in the BRH material, then, gives the amount of PCBs in the SPM in

the organism's guts.


Fe concentration Day 28 Mussels        Fe concentration Control Mussels
     500 ug/g dry wt mussel       -    193 ug/g dry wt mussel
        (Table 7)                            (Table 7)

                                  equals

                              Difference 307
                              ug/g dry wt mussel


If the assumption is made that all the Fe in the mussel at day 28 results

from Fe on the SPM in the gut of the organism,


(307 ug Fe/g dry wt mussel) (1000 mg dry wt BRH sed./29600 ug/Fe)
                                   (Table 17)

                                  equals

                  (10 mg dry wt BRH sed/g dry wt mussel)


(10 mg dry wt BRH sed/g  dry wt mussel)  (6800 ng A-1254/1000  mg dry
                                           dry wt.  BRH sed)*
                                  equals

                         (71 ng A-1254/g dry wt mussel)

Since  the increase  observed in the concentration of PCBs  in  the  mussels

is much larger  than this,  28 day mussel - 2800 ng/g (Figure  12), the

contribution  from PCBs on  SPM  in the gut of the organisms appears to be

almost inconsequential.

       64.   The  concentrations  of PCB in mussels exposed for  28 days were

divided by  the  concentrations  of PCBs  in  filtered  water samples  to obtain

bioconcentration factors (BCFs).**   These data  expressed as  Log  BCF

  *  Value  from Rogerson et  al.  (1983).
**  Bioconcentration in  this  report refers to  the process  of  uptake
    of  contaminants  from water.
                                     55

-------
                          Table 7
  Average Trace Metal Concentrations for Mussels collected

Metal
Fe
Zn
Mn
Cu
Pb
Cd
Cr
from the Exposure

Chamber on Day 28*

Mussel
28 Day
500 +
333 +
11 +
55 +
13.9 +
7.0 +
25.1 +
191
84
5
18
4.7
2.0
10.7

Mussel
Control
193 + 24
178 + 53
12+5
12+5
5.0 + 1.5
2.6 + 0.4
2.2 + 1.0
*The control concentrations reported for the mussels are the
 average of all the control samples and not just day 28.  All
 concentrations are in Ug/g dry weight.
                             56

-------
(Table 8) were converted from dry wt to wet wt using a common wet to dry

conversion factor to facilitate comparisons with estimates of Log BCFs

from Geyer et al.(1982).  Due to variability in the amount of water in

the organism tissues, the authors prefer the use of dry weights from

individual samples for calculations, as is done in the remainder of the

report.  The measured Log BCFs for representative PCB compounds increase

with increasing Log P (decreasing aqueous solubility) as observed for

Log BCFs with mussels (Ernst 1977) and fish (Veith et al. 1979).  In

addition, the measured values are in close agreement with estimated Log

BCFs (Geyer et al. 1982).

      65.  The concentrations of compounds in the mussel samples at day

28 (dry weight) and the concentrations of compounds in the unfiltered

water samples at day 28 were used to calculate bioaccumulation factors

(BAFs).*

     BAF =« concentration of individual PCB compound in mussel (dry weight)
           concentration of individual PCB compound in unfiltered water

In order to facilitate comparisons of these large values, log BAFs were

calculated (Table 9).  In spite of considerable differences in the n-

octanol/water partition coefficients (Log Ps) for these PCB compounds,

the Log BAFs appear to be quite constant.  This is in contrast to BCFs

(accumulation from water only) from the literature for single compound

tests with dissolved components, which show increasing BCFs with
                                    57

-------
  1000
T3


O»
\
O>
CVJ

<


CO
o
0_
   100
     10
                                                 exposure
                                                  tank
                                                 control
                                                  tank
                   UPTAKE
                                   DEPURATION
       1.83.57    14    21     28   35 40     49    56    63   70

                                TIME  (days)


       Figure 12.  Concentration of total PCBs (as  A-1254)  in mussels
                   exposed to BRH sediment versus time
                                 58

-------
                                      Table 8

              Estimated  and Measured Bioconcentration Factors (BCF) in
Mussels at Day 28




Log P* Estimated Log BCF**
Peak No. PCB Compound (Log Row) (wet wt.)
4 2, 2 ',5 - trichlorobiphenyl
10 2,4,4', -
25 2,2'4,5,5
36 2, 2', 4, 4'
43 2, 2', 3, 3'
PAHs
Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benz(a)anthracene
Chrysene
Benz(a)pyrene
Perylene
trichlorobiphenyl
'-pentachlorobiphenyl
,5,5' -hexachlorobiphenyl
,4,4' -hexachlorobiphenyl









4.7
5.0
6.3
6.7
7.0

4.4
5.3
4.9
5.1
5.8
6.4
6.2
6.9
3.2
3.5
4.6
4.9
5.2

3.0
3.7
3.4
3.6
4.2
4.7
4.5
5.1

Measured Logt BCF
(wet wt.)
3.2
3.6
4.2
4.7
4.8

2.2
2.3
2.9
3.1
4.0
3.9
4.2
3.8
 * PCB solubilities from Mackay et al. (1980a).   PAH solubilities from
   Mackay et al. (1980b).   Solubility converted to Log P using Log P -
   5.00 - .67 Log S where S is solubility in umol/L (Chiou et al. 1977.)

** BCF estimated from log BF - 0.858 x Log Row - 0.808 from Geyer et al.(1982),
   (BCF can be substituted for BF.)

 t BCF measured from mean of concentrations in three 28-day exposed mussels
   divided by mean of concentrations in three 28-day filtered water samples
   from the exposed tank.


                                      59

-------
 decreasing water solubility (increasing Log P) for mussels (Ernst 1977;




 Geyer et al.  1982).   The uniform BAFs observed in the present study




 probably resulted from the presence of SPM in the dosing system.




       66.   The constant BAFs  observed may have resulted from two




 processes  competing  for the dissolved phase contaminants.  The first is




 re-adsorption  of dissolved PCB  contaminants by the SPM including  algae; the




 second  is  the  bioconcentration  of  dissolved PCB contaminants by the mussels.




 If  these two distributions vary to approximately the  same extent  over the




 range of PCB contaminants, then constancy of BAFs  could result.  Another




 possible explanation for the  relatively constant BAFs  observed in this




 study is that  the mussels  accumulate  individual PCB compounds by  a similar




 constant process (i.e.,  transfer from particles across the  lining of  the




 gut).   This method of  accumulation could result in distributions  which




 were very  similar to  those in the  unfiltered water and filter samples if




 depuration rates for  the individual compounds  were approximately  equal




 during  accumulation.   A third possible  explanation for the  constant BAFs




 observed in this study is  that  steady-state  values were not  reached for




 all PCB  compounds during the uptake period  (see discussion  of  Kinetics).




      67.  The distributions of dissolved  (filtrate) and  particle-bound




 (filter) PCBs were compared with those  in mussels  (Figure 13).  The




distribution in  the water  (filtrate) is  dominated  by low molecular weight




compounds and the peak heights of  the PCBs decrease with increasing




molecular weight.  The distribution on  the SPM  (filter) closely matches




that found in mussels  (Figure 14), suggesting that  PCBs in the  SPM




influence the distribution found in the mussels.
                                    60

-------
                                Table 9
    Measured Log BAF* for Each Separate PCB Peak in Exposed Mussels
Peak Number
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
Log BAF
4.3
4.4
4.5
4.4
4.4
4.1
4.4
4.6
4.5
4.4
4.4
4.4
4.4
4.4
4.5
4.5
4.4
4.4
4.5
4.5
4.4
4.4
4.4
4.5
4.4
4.4
4.4
Peak Number
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51



Log BAF
4.5
4.6
4.5
4.5
4.5
4.6
4.5
4.5
4.5
4.5
3.9
4.4
4.2
4.4
4.4
4.4
3.4
4.3
4.2
4.4
4.1
3.0
3.3
3.8



*Method for calculation of Log BAF shown in text,
                                  61

-------
                   1
                                       Time 	>

                             a.  Filter, day
                                        Tim«
                             b.   Filtrate,  day
                           c.  Mussels, day 28


Figure 13.  Capillary column electron capture gas chromatograms of PF-50
                       (PCB) fraction from exposure
                                    62

-------
      68.  Chromatograms from control mussels sampled at day 0,




day 28, and day 70 (Figure 15) showed only minor changes between day 0



and day 28.  Between day 28 and day 70 an Increase In the height of the



2,4,8-trichlorodibenzofuran and tetrachlorodiphenyl ether peak was




evident in the control mussels.  This increase probably reflects an




increased input of these industrial contaminants to the upper




Narragansett Bay followed by down Bay transport and entrance of small



amounts of these contaminants into our laboratory seawater supply.  In



addition, a small relative increase in some lower molecular weight PCB




compounds was observed in the day 70 control.  During the depuration




period, a late eluting peak appeared in chromatograms.  GC/MS analysis



showed that it was not a chlorine or bromine-containing compound•  It is




probably an electron capturing biological compound.  It should be noted




that the PCB concentrations in control mussels remained low during the




experiment and that these organisms fulfilled their intended purpose as




chemical controls by accumulating background concentrations of pollutants



from the control seawater.




      69.  PCB Kinetics.  The accumulation and depuration of PCB




contaminants (quantified as Aroclor®-1254) are shown in Figure 12.  To




determine if steady-state was reached during the uptake period, the



A-1254 residue concentration in the mussels, and the time data (in days),




were entered into a computerized non-linear model in accordance with




proposed ASTM recommendations (ASTM 1982).






          Y - Ln  Residue - Pl/(l + P2 ** (Time - P3))
                                    63

-------
where Y « natural log of the residue
      Pl= natural log of the maximum predicted residue concentration
      P2= rising slope (0
-------
                    NUMBERS ABOVE PEAKS REFER TO IDENTITIES

                    OF COMPOUNDS LISTED IN TABLE 5.
   I
   :.
   in
   -a
   :•
   o


   n

   v

   r i
                                       Time 	>




                               a.  Filter,  day 8
                                f1
Figure  14
                          Time	>



                  b.  Mussels,  day 28





Capillary  column electron  capture gas chroraatograms of  PF-50


            (PCB) fraction  from exposure
                                      65

-------
                         NUMBERS ABOVE PEAKS REFER TO IDENTITIES
                         OF COMPOUNDS LISTED IN TABLE 5
                              b.   Control,  day 28
                             c.
       Time

Control,  day 70
Figure  15.   Capillary column  electron capture gas  chromatograms  of PF-50
                         (PCB)  fractions  from mussels
                                       66

-------
      73.  The kinetics of the depuration phase were examined in detail.




For most bioaccumulatlon studies, first-order kinetic expressions have




been applied (Niimi and Cho 1981; Ernst 1977; Veith et al. 1979).  For



this process, the depuration rate is not dependent on the initial



concentration.  Another study found that' second order kinetics were




followed for the elimination of pesticides from catfish (Ellgehausen et



al. 1980).  In second-order processes the depuration rate is dependent




on the initial concentration.  Plots of In C versus time (C=*



concentration) and 1/C versus time for all the peaks examined during the




depuration phase were made.  If first-order kinetics were followed, then




the plot of In C versus time should be linear; if second-order kinetics



were followed, the plot of 1/C versus time should be linear  (Glasstone



and Lewis 1960; Ellgehausen et al. 1980).  No clear distinction of the




order of the kinetics was found in comparisons of the correlation co-




efficients (Table Al).  In addition, scatter of the data during depuration



and the impact of slightly elevated levels of PCBs in the exposure tank



during depuration (Table 2) precluded a conclusive determination of the



order of kinetics.




      74.  If first-order kinetics are assumed, as was the case in other



studies on bioaccumulatlon (Niimi and Cho  1981; Ernst 1977;  Vieth et al.




1979), differences in  the depuration rates for the accumulated compounds



can be examined.  The  slopes of lines  (the first-order depuration rates)




for the compounds examined in this study are shown on Table  A2.  A subset




of eight  "representative" compounds was selected  from the PCB dis-



tributions in mussels  for more detailed study.  Analysis of  covariance



was used  to  test equality of the slopes for the eight compounds  (ot. =»0.05)
                                     67

-------
over different sections of the depuration period.  This examination was




made over different depuration periods  (1 week,  2 weeks,  3 weeks,  4



weeks, 5 weeks, and 6 weeks) to determine if the depuration rates  were



constant over the depuration period.  The results (Table  A2) show  a




decrease in depuration rate (demonstrated as less negative slopes) with




increasing depuration time for most individual compounds.  These results




may demonstrate the inapplicability of  first-order kinetics to describe



depuration for some of these compounds.  The results of the comparisons




of the equality of the slopes (depuration rates) during the depuration




time periods are shown in Table A2.  The results show that statistical




differences between some lines exist (a =0.05) over some  of the time



periods.  Within each depuration period there are 28 possible compound




comparisons.  Consequently, a very small a level (.05/28) was chosen for




each pairwise comparison.  This was done to maintain a 5% level of




significance for all pairwise comparisons within each depuration period.




While differences between other lines were not significant at the  same



concentration, an observed trend showed that the lower molecular weight




compounds were more rapidly depurated than the higher molecular weight



compounds.  Some higher molecular weight compounds appear to depurate




faster than some mid range PCBs during  the first weeks of depuration (up



to day 56 or 28 days depuration).  The slopes of the depuration lines



for the different compounds converge as depuration time increases.




      75.  If all the depuration data are included, those compounds




which are resistant to transformation (Zell and Ballschmiter 1980) and



with higher chlorination have the slowest depuration rates (shown as



less negative slopes in Table A2).
                                   68

-------
      76.  PAHs - Seawater.  The concentrations of 11 polycyclic aromatic




hydrocarbons (PAHs) and one chlorinated pesticide, Ethylan (1,l-dichloro-2,




2-bis (p-ethylphenyl) ethane), in unfiltered water samples (dissolved plus



particle-bound compounds) taken during the exposure phase of the mussel




bioaccumulation study are shown in Table 10.  The levels of these




contaminants in control water samples, water samples taken during




depuration, and blanks were below the detection limit «0.1 ng/L for the




methods used for extraction and analysis).



      77.  Extracted ion current profiles  (EICPs) result from the GC/MS




analysis.  These profiles display the concentrations of the major ion




for each compound as a function of retention time on the GC column.



By examining several of these plots corresponding to different  times




during the course of the experiment it is  possible to determine what




relative changes in the content of selected compounds occurred  during




the experiment.




      78.  The EICPs for the  PAH and  Ethylan compounds  (which are



reported together because  they were all  analyzed  in  the same  GC/MS



analyses)  in  an unfiltered exposure water  sample  from the  dosing system




day  8 are  shown in Figure  16.   The mass  numbers  (molecular weight/charge)




of  fragments  characteristic of  the compounds  (Figure 16)  are  shown  on




the  right  axis.



       79.   Examination of  the unfiltered water sample  EICP (Figure  16)




and the  data in  Table  10 indicates that  the relative distributions  of PAH




 compounds  and the Ethylan were fairly consistent over  the exposure  studies,



 but the  total concentrations  of these compounds changed (RSDs (S.D./mean




 x 100)  for PAH compounds were up to  approximately 75%).  The greater
                                     69

-------
                                    Table  10
            PAH and Ethylan Concentrations in Unfiltered Water  Samples*
Day

Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benz(a)anthracene
Chrysene
Benzo(b)f luoranthene and/or
Benzo(k)fluoranthene
Benzo (e )pyrene
Benzo(a)pyrene
Perylene
Sum of PAHs with MW of 276
Ethylan
SUM-PAHs
£
6.8
2.3
17.0
25.9
13.7
20.1
25.4

15.7
16.1
2.9
25.2
0.8
171.
8_
39.6
9.6
47.8
74.3
33.2
45.5
59.3

34.6
35.9
6.4
57.8
1.6
444.
J.4
8.2
2.1
11.5
18.5
8.7
13.1
16.8

9.4
9.7
1.9
16.8
0.6
117.
28
28.1
9.2
33.5
49.5
19.'
29.0
36.7

19.7
22.3
4.3
38.*
1.2,
290.
*  (in Parts per Trillion)
                                  70

-------
           LETTERS ABOVE PEAKS REFER TO IDENTITIES
           OF COMPOUNDS LISTED IN TABLE 11.
A

-
B C
-- 	


D
J




uu-~
L E


--
F G


1
H I J K
I I MASS
1 AL/L
1 -— 276
L (
	 — f^e.
\ A -
i- — . — ~ — . — _____ — . — . — — _ 223
•• 	 —202
.
i [ i | • |~ i |- \FB
1200 1400 1600 1800 2000 2200 SCAN
2000 23*20 26*40 30*00 33*20 36*40 TIME
MAP:
a. Unfiltered water
A

B C
,1.
D L
-J
-*-*-
E
.._J
	
F G H
-
I J K
MASS
	 - - --276
CJi
	 — — 	 .— — -V— 	 _— ~- _ ^.jj

1 | 1 | l | l | 1 | l
1200 1400 !600 1800 2000 2200 SCAN
20*00 23*20 26*40 30*00 33*20 36*40 TIME
MAP*
                 c.   Filtrate
                                                                  A B
                                                                          C D L
                                                                                  E F
                                                                                         GHIJ
                                                                       ._JJUL
                                  — 223

                                  -— 202
                                                                    '2°°
                                                                    20*00
I40°
23*20
 I6O°
26*40
 I80°
30*00
200°
36*20
2200 SCAN
36*40 TIME
                                                                MAP:
                                                                  AB
                                                                                     b.   Filter
                                                                          COL
                                                                                  E F
                                                                                         G HIJ
                                                                    1200
                                                                    20*OO
 1400
23*20
                                                                MAP:
 1600
26*40
 1800
30*00
2000
33*20
                                                                                                              MASS

                                                                                                             • - 302


                                                                                                             •- 276


                                                                                                             -- 252


                                                                                                             •- 223
    178
2200 SCAN
36*40 TIME
                                                                        d.  Water  through  continuous  flow
                                                                                  centrifuge
                       Figure  16.   EICPs from GC/MS analysis of  exposure  tank

-------
                               Table 11
                       PAH Compounds in Mussels
Peak                 Chemical ID




 A                   Phenanthrene



 B                   Anthracene




 C                   Fluoranthene




 D                   Pyrene



 E                   Benz(a)anthracene




 F                   Chrysene




 G                   Benzo(b)fluoranthene and/or Benzo(k)fluoranthene




 H                   Benzo(e)pyrene




 I                   Benzo(a)pyrene




 J                   Perylene




 K                   Sum of PAHs with MW of 276




 L                   Ethylan
                                72

-------
 variability observed for the PAH compounds in water samples than for the

 PCB compounds (paragraph 54) may reflect variability of  the contaminants

 in the BRH dredged material.  It should be noted that soot  particles

 containing high concentrations of PAH compounds  may be present  in

 contaminated sediments and that variability in the  numbers  of these

 particles  in samples may substantially contribute to concentration

 variability.

       80.   The  EICPs from the GC/MS  analyses  of  unfiltered  water, filters,

 filtrate,  and water passing  through  the  continuous  flow  centrifuge

 taken  on day 8  of  exposure are shown in  Figure 16.   The  samples  of un-

 filtered water  show a pattern of  peaks for  the compounds of  interest

 which  is very similar to the patterns  for  the BRH sediment.  A similar

 distribution is observed in  the sample from the  filter.  The EICPs from

 the filtrate and the water passing through  the continuous flow centrifuge

 show a relative enhancement  of  the lower molecular weight PAH compounds.

 As found with the  PCB compounds,  the PAH compounds appear to distribute

 in accordance with  their  solubilities.  With  lower molecular weight, more

 soluble PAH  compounds  are  found in the filtrate, and with the higher

molecular weight, less soluble compounds are  found associated with

particles.

      81.  Data from  the analysis of filtered material and filtrates were

used to calculate sediment-water partition coefficients,  Kp, where

                       Kp»Cs/Cw

                Cs - concentration of compound in sediment (dry weight)
                Cw » concentration of compound In water

Kps were estimated for compounds where the Log n-octanol/water partition
                                    73

-------
coefficient (Log P) values were known.  As observed for the PCB




compounds, the estimated results for the PAHs were considerably below




the experimental results (Table 12).  This may indicate that the de-




sorption of PAH compounds from suspended sediment was not complete (i.e.,




equilibrium was not reached) during the residence time of the suspensions




in the dosing system.



      82.  PAHs - Mussels.  Examination of the EICPs from mussels at




day 0 showed phenanthrene, fluoranthene and pyrene, benz[a]anthracene,




benzo[k] and/or benzo[b]fluoranthene, benzo(e]pyrene, and benzo[a]pyrene




and perylene (Figure 17).  Ethylan was not found in these background




samples.  At the first sampling period (day 1.8), the abundance of the




above compounds had increased and anthracene, Ethylan, and some PAHs




with MW 276 were apparent (Figure 17).  Comparison of this EICP with one




from the BRH material shows that mussels had a relatively lower concen-




tration of peaks G, H, I, J, and K than was present in the sediment (Figure




17).  This same general pattern of peaks is observed in all other mussel




samples taken during exposure (Figures 17 and 18).  Following 7 days of




depuration, the lower molecular weight peaks A, B, C, and D had decreased




considerably while peaks E through K had become more prominent (Figure




19).  The selective depuration of the lower molecular weight peaks may




result from the higher depuration rates associated with more water-soluble




compounds (Ernst 1977).  As depuration continued, general decreases in




the concentrations of all compounds were observed (Figures 19 and 20).

-------
                                 Table 12
          Comparison of Experimental and Estimated Sediment/Water
          "Partition Coefficients (KpsT
PAH Compound
Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benz(a)anthracene
Chrysene
Benz(a)pyrene
Perylene
Log P*
4.4
5.3
4.9
5.1
5.8
6.4
6.2
6.9
Kp Experimental**
.27 X 105
.23 X 105
.71 X 105
.79 X 105
4.7 X 105
4.1 X 105
24. X 105
17. X 105
Kp Estimatedt
.009 X 105
.066 X 105
.048 X 105
.031 X 105
.23 X 105
.87 X 105
.60 X 105
2.8 X 105
* Solubility from Mackay et al. (1980a) converted to Log P using
  Log P = 5.00 .67 Log S where S is solubility in ymol/L (Chiou et al.
  1977).

**Kp estimated as mean of 3 Kps determined on day 28 from mussel exposure
  tank.

t Kp estimated as in Appendix Table A-l.
                                    75

-------
~~J
              A B
                      LETTERS ABOVE PEAKS REFER TO IDENTITIES
                      OF COMPOUNDS LISTED IN TABLE 11.
                      C D  L     £F     GH]  J     K




MA
A








_J
1
1200
20:00
P:
B C









JL
c



«.
.







,


--, i
1
k -
1 f 1 '
LJli 	
r
L .
' - i ii -

._j , .
i
2
) I



1
400
3^20
a.
E


--JL
1
1600
26:40
BRH 5
F G


I
1800
30:00
iedimer
HIJ K
1 i —
i -4_
L -





MASS
— 302


— 223
— 202
1 | I .10
2000 SCAN
33:20 TIME
its
MASS
— i




I ' ' 1 ' 1 '
1200 I40O I60O 1800
20:00 23:20 26:40 30:00

2OOO 22
33:20 36


— 252

— 202
00 SCAN
40 TIME
            MAP:
                c.  Exposure  tank  mussels,  day  l.i
\B (
-


	 J-« — f.

:


4*
^
i.
D





E
-• ---•

	 ,_


F


i
J
|
c
	

bd'j-
J,
|
H
"j
1



Ivl 1
J
1 — '



,.
;
— i
i



nn » .^i .

MASS
— 302
— 276
— 252
— 223
— 2O2
                                                                                    1200
                                                                                   20-00
         I40O
        23^20
 1600
26^40
 1800
30^00
2000
33^20
SCAN
TIME
                                                                               MAP:
                                                                                AS
                                                                                                   Mussels, day 0
                                                                                       C D L
                                                                                                EF
                                                                                                      G HIJ     K




i
. .. LrfJ
l,__ 	 -j .--M




fl




CC




__L



^


U
j
...
I

MASS
— i


•-— ri^" A, , , - -

r i |

i | i -
— OUi
— 276
— 252
-223
— 202
— 178
                                                                              MAP:
                                                                                  1200     1400     1600     1800    200O    2200 SCAN
                                                                                 20:00     23:20    26:40     30:00    33:20    36:40 TIME
d.   Exposure  tank mussels,  day  3.5
                                               Figure  17.   EICPs  from GC/MS  analysis

-------
A B
         LETTERS ABOVE PEAKS REFER TO IDENTITIES
         OF COMPOUNDS LISTED IN TABLE 11.
        CDL     EF     GHIJ     K
                                                                              CDL
                                                                                       EF
                                                                                              G HIJ




	 JL.."-« —
r '
J


t






*




	 L










\



V
' '



'I' ""i 	 I"*" "l 	
                                               223
    1200    1400     1600     1800     2000
   20>00    23*20    26*40    30^00    33*20

MAP

                     a.   Day 7


AB     CDL    EF     GHIJ    K
2200
36-40





:



i



J



__L



L


*-.
*
-'




-T*""*T r 1
1200 1400
20*00 23*20
II 1
1600 1800 2000
26*40 30*00 33*20
                                                276


                                                252



                                                223


                                                202


                                                178

                                                SCAN
                                                TIME
 MAP:
	

t
f* —
12
JL-J.



*


^




-JL



L


p*""1" i
00 1400 16
I



LJ

i 	 '

	 1
A— -. -




rfASS
- 302
— 276
252

- 202
— 178
1 1 ' 1 """
00 1800 2000 SCA
                                                                     MAP:
                                                                                         b.   Day  14
                                                                      AB      CDL
                                                                                       EF
                                                                                              GHIJ
                                                                     MAP:
                                                                                                                     T1ME




__JLJV
\M~^*I~**-
,



A



DC
J
~V
-------
             AB
                      LETTERS ABOVE PEAKS REFER TO IDENTITIES
                      OF COMPOUNDS LISTED IN TABLE 11
                     C D L    EF
                                   G HIJ


*M
MB




c=A—




X
h
JU



J



«^A.
•TT ., i



bzs~
ttaM


k
***

\ 	
_j.
r it
I BlMTII

1 '1 1
1200 1400 1600 1800
20=00 23=20 26=40 30=00
MASS
1 	 1
II

•BH^IMMAM


— 276
-252
— 223
- 202
— r— 3- 178
2000 SCAN
33=20 TIME
            MAP
                               a.   Day 35
                                                                          MAP
                                                                                             b.  Day 40
                                                                                                                    MASS
00
                                                                                   COL   E F
                                                                                                 G H IJ
                             c.   Day  49


— h
1 ._.*



•f
Ik
«-
\M


Vv






***V*t
i
1200 1400
20=00 23=20






'
7
i —
MASS
1 	 1
-. 	 *<..„.,
. .
L*-^1" ill i.i _._•>.. j
^L. _t.tu ^d**^''ft»**»««»»M


— 252
— 223
— 202
I7Q
1 1 . 1 ' 1 1
1600 1800 2000 2200 SCAN
26=40 30-00 33=20 36=40 TIME
                                                                          MAP:
                                                                                           d.   Day  56
                             Figure 19.   EICPs from  GC/MS  analysis of exposure  tank mussels

-------
                  LETTERS ABOVE PEAKS REFER TO IDENTITIES
                  OF COMPOUNDS LISTED IN TABLE 11
                C D  L     E F      GHIJ      K
          I200
         20'00
I400
23^20
 1600
26^40
 1800
30-00
2000
3 3'20
     MAP:
                                                             178
 2200 SCAN
36'40 TIME
         y-  —r    i     i     |     ,     |	1	1	1	\~ 178
         1200       1400      1600      1800      2000      2200  SCAN
        20:00      23^20     26 = 40     30^00      33 = 20      36:40  TIME
    MAP:

                              b.  Day 70

Figure  20.   EICPs from  GC/MS analysis of  exposure  tank mussels
                                 79

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                                      Table  13


              Estimated and Measured  Bioconcentration Factors  (BCF) in

Mussels at Day

Log P*
Peak No. PCB Compound (Log Kow)
4 2, 2', 5 -
10 2,4,4', -
25 2,2-4,5,5
36 2, 2', 4, 4'
43 2, 2', 3, 3'
PAHs
Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benz ( a ) anthrace ne
Chrysene
Benz(a)pyrene
Perylene
trichlorobiphenyl 4.7
trichloroblphenyl 5.0
'-pentachlorobiphenyl 6.3
,5,5'-hexachloroblphenyl 6.7
,4,4'-hexachloroblphenyl 7.0
4.4
5.3
4.9
5.1
5.8
6.4
6.2
6.9
28

Estimated Log BCF**
(wet wt.)
3.2
3.5
4.6
4.9
5.2
3.0
3.7
3.4
3.6
4.2
4.7
4.5
5.1

Measured Log! BCF
(wet wt.)
3.2
3.6
4.2
4.7
4.8
2.2
2.3
2.9
3.1
4.0
3.9
4.2
3.8
 * PCB solubilities from Mackay et al. (1980b).  PAH solubilities from
   Mackay et al. (1980a).  Solubility converted to Log P using Log P -
   5.00 - .67 Log S where S is solubility in jjmol/L (Chiou et al., 1977)

** BCF estimated from log BF - 0.858 x Log Kow - 0.808 from Geyer et al.
   (1982).

 t BCF measured from mean of concentrations in three 28-day exposed mussels
   divided by mean of concentrations in three 28-day filtered water samples
   from the exposed tank.
                                          80

-------
      83.   The mean concentrations of PAH compounds in mussels exposed




 for 28 days were divided by the concentrations of PAHs in filtered




 seawater  samples to obtain bioconcentration factors (BCFs) (Table 13).




 As  with the PCBs (paragraph 64) these data are expressed in Log form on




 a wet weight basis to facilitate comparisons with estimates of Log BCFs




 from Geyer et al.  (1982).   The  measured  Log BCFs  for the PAH compounds




 increase  with increasing Log P  (decreasing aqueous solubility) as observed




 for Log BCFs with mussels  (Ernst 1977) and fish (Veith et al. 1979).




 For PAHs  the measured values are not  as  close to  the estimated values




 as  were the PCBs (Table  13).




       84.   The mean concentration of  PAH and Ethylan compounds in the




 mussels at  day 28  (Table 14) and the  mean concentration of these  compounds




 in  unfiltered water at day 28 (Table  10) were used to calculate bio-




 accumulation factors  (BAFs). The  Log BAFs and the Log Ps are shown  in




 Table  15.   The compounds with lower Log  Ps showed  lower BAFs.   Benz(a)




 anthracene  and chrysene  showed  the highest BAF values in the mussels




 while  the higher molecular weight  PAH compounds were accumulated  less




 effectively.   The pesticide  Ethylan showed a relatively high BAF  compared




 to  the  PAH  compounds.  Since organisms were not gut  depurated prior  to




 analysis, the  PAH content  of the organisms included  a contribution from




 sediment in the  gut  (see discussion under  PCBs, paragraph 63).  The




 levels of PAH contaminants  found in control  mussels  were  low and  remained




 relatively  constant during  the dosing-period.   Ethylan was not  found in




 control samples  (Table 16).



      85.   The uptake  and depuration of  total  PAH  compounds  during the




mussel exposure  study  are shown in Figure  21.   This  plot  was made using
                                    81

-------
00
ro
                                                               Table  14


                             PAH and Ethylan Concentrations  in Exposed Mussels Expressed as ng/g (dry)*

Day
Peak
Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benz ( a )anthracene
Chrysene
0
8.69
.488
17.4
15.8
2.72
6.63
Benzo(b)fluoranthene 9.82
and/or Benzo(k)fluoranthene
Benzo(e)pyrene
Benzo(a)pyrene
Perylene
Sum of PAHs with
MW of 276
Ethyl an
Sum of PAH Compounds

3.96
.532
1.12
3.66
0
70.8
7
162.
63.6
802
1359
754
1005
631
288
269
30.7
261
452.
5630
Expos ur
14
130.
49.6
444
811
448
650
408
234
216
25.5
170
177.
3590

e~
21
246.
73.0
475
1117
512
856
543
301
350
60.3
111
317.
4640

>_ „
™
28
130.
59.8
698
1228
864
1179
895
379
392
44.0
348
444.
6220

35
10.7
4.52
59.1
175
285
435
472
249
226
23.9
141
142.
2080
— — Tlav
itef
40
13.6
4.81
37.6
95.3
153
270
288
161
120
18.4
82.7
154.
1250
>uration
49
10.1
2.75
19.8
38.8
35.9
75.4
101
59.0
31.1
4.30
21.4
29.0
399

56
23.2
5.43
17.9
35.0
9.76
25.2
29.0
22.6
7.01
4.13
7.57
14.5
187

62
9.65
2.17
23.4
41.7
22.8
40.6
73.1
23.5
16.1
4.26
20.6
20.3
278

70
10.9
2.57
20.1
33.3
4.99
16.0
12.8
0
0
0
5.97
8.8
107
         * Values not corrected for blank  value.

-------
                                 Table  15


           Mussel Bioaccumulation Factors  (Calculated for  Day  28)
Peak
Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benz(a)anthracene
Chrysene
Benzo(b)fluoranthene and/or
Benzo(k)fluoranthene
Benzo(e)pyrene
Benzo(a)pyrene
Perylene
Sum of PAHs with MW of 276
Ethyl an
Log BAF
3.7
3.8
4.3
4.4
4.7
4.6
4.4
4.3
4.2
4.0
4.0
4.6
Log P*
4.4
5.3
4.9
5.1
5.8
6.4


6.2
6.9
7.0

* P » n-octanol/water partition coefficient obtained from solubility
  data in (Mackay et al. 1980a).  Converted to Log P using
  Log P « 5.00-.67 log S where S is solubility in umol/1 (Chiou et
  al. 1977).
                                   83

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                                 Table  16
           Levels of PAH and Ethylan Compounds  in  Control  Mussels
During


Peak
Phenanthrene
Anthracene
Fluor anthene
Pyrene
Benz ( a )anthracene
Chrysene
Benzo(b)£luoranthene and/or
Benzo(k)f luoranthene
Benzo(e)pyrene
Benzo(a)pyrene
Perylene
Sum of PAHs with MW of 276
Ethylan
Study (PPb;


0
8.69
.488
17.4
15.8
2.72
6.63
9.82
3.96
.532
1.12
3.66
0
(ng/g(dry))

Day
ire — ~ —x Depuration
28
6.36 5
.697 1
6.12 11
9.31 13
1.13
3.12 7
3.84 1
2.07 4
.621
.355
2.28 2
0



56
.70
.20
.7
.6
.852
.95
.62
.56
.399
.221
.01
0


- _ _ _S
70
6.22
.383
7.78
13.4
.911
5.80
2.14
3.92
.075
.075
.524
0
Sum-PAHs
70.8
35.9
49.8
41.2
                                        84

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CO
Q.
Q.
                                                                EXPOSED
                                                            REFERENCE
                                        DEPURATION
f UPTAKE
       |	1	1	1	1	1	1	1     I    I    I	1"	1	1    I
       05    10  15   20   25  30 35  40  45  50   55   60  65  70
                                       DAYS


     Figure  21.   Concentration of sum of parent PAHs in mussels exposed
                         to BRH sediment versus time
                                      85

-------
the sums of  the concentrations of the eleven  PAH  compounds  listed  in




Table  14 and shows a rapid uptake of PAH compounds to day 7 of  the exposure,




Following day  7 concentrations fluctuated until the end of  the  exposure




at day 28.   Depuration of the PAH compounds was rapid during the first




week followed  by several weeks of slower depuration.  By day 70 the




concentrations of most compounds had decreased to approximately two to




three times  their day 0 levels indicating that depuration was not




complete.




      86.  Figures A18 to A20 show the uptake and depuration of the




PAHs and the Ethylan.  Most of the curves showed maximums at day 7 or




day 28.  The curves for fluoranthene and pyrene showed maximum




concentrations at day 7, but in general larger PAH compounds showed




maximum concentrations at day 28.




      87.  During the depuration phase, the lower molecular weight PAH




compounds appeared to be depurated more rapidly that the higher molecular




weight PAH compounds.  The rapid depuration of more soluble (lower Log




P) compounds by mussels has been observed in another study  (Ernst  1977).




In general, the higher molecular weight PAH compounds (higher Log P)




took longer to reach their maximum level and were depurated more slowly




than the lower molecular weight (lower Log P) compounds.  Ethylan which




was not detected in control organisms was accumulated and depurated similar




to chrysene.




      88.  Petroleum Hydrocarbons - Mussels.  The petroleum hydrocarbons




from samples were detected as a large mound in flame ionization detection



gas chromatograms (Figure 22).  This mound of material, usually referred




to as an unresolved complex mixture (UCM), consists of numerous petroleum
                                    86

-------
              a.  Exposure tank water, day 0
                       b.   BRH sediment
               c.   Mussels,  exposute day 28
Figure 22.  Capillary column flame ionization detector gas
chromatograms of PF-50 (contains mostly straight chain, branched,
           and cyclic saturated hydrocarbons)

-------
hydrocarbons (i.e., alkanes, cycloalkanes).  The petroleum hydrocarbons




found In unfiltered water samples from the dosing system showed a




distribution as an unresolved complex mixture (UCM) which was slightly




lower in molecular weight, but otherwise similar to the distribution




found in the BRH sediments (Figure 22).  During the uptake phase the




mussels accumulated a UCM which was slightly lower in molecular weight




than the distribution found in the unfiltered water.




      89.  The petroleum hydrocarbons followed the same general pattern




of concentration changes observed during the uptake and depuration for




the other organic contaminants in mussels (Figure 23).  By day 7 the



contaminants were near their maximum values.  At day 14 and day 21 lower




concentrations were observed with the highest concentration found at day




28.  The first week of depuration showed a rapid loss of total petroleum




hydrocarbon contaminants followed by a plateau phase of decreased loss




rates.  This behavior for the depuration of petroleum hydrocarbons has




been observed in other studies with bivalve molluscs (Lee et al. 1972;




Clark and Finley 1975; Fossato 1975; Lake and Hershner 1977).  Chromato-




grams from control mussels showed a low level of petroleum hydrocarbons




(as a UCM).  Petroleum hydrocarbons in mussels from the control tank




showed slight concentration decreases during the study period (Figure




23).






Inorganic Contaminants




      90.  Sediment.  The trace metal composition for the barrel of BRH




sediment analyzed is given in Table 17.  The wet-to-dry-weight
                                    88

-------
   500 r
£
O
10
                                                    exposure

                                                      tank
                                                    control
                                                      tank
 Figure 23.
                  UPTAKE	h
                               DEPURATION
              10      20      30      40

                              TIME  (days)
                                           50
60
70
       Concentration of total petroleum hydrocarbons in mussels

             exposed to BRH sediment versus time
                                89

-------
                            Table  17
    Average Trace Metal  Concentrations  for Black Rock Harbor
          Sediment  Samples  Expressed as yg/g Dry Weight
   Metal          pg/g           Std.  Dev.            % Std. Dev.

    Fe            29600             809                    2

    Zn            1200              59                    4

    Mn             359              37                   10

    Cu            2380             112                    4

    Pb             378              16                    4

    Cd               23.4             0.9                   4

    Cr            1430              77                    5

    Ni             139               4                    3

    Hg               1.7             0.1                   4


Wet/Dry              3.22            0.02                  0.6
                              90

-------
                               Table 18
   Seavater Metal Concentrations Determined for the Black Rock Harbor
      Sediment Exposure and Control Chambers Expressed as ug/liter
Metal
Fe
Std. Dev.
Zn
Std. Dev.
Mn
Std. Dev.
Cu
Std. Dev.
Pb
Std. Dev.
Cd
Std. Dev.
Cr
Std. Dev.
Exposure
Pre renewal
of BRH Slurry
288
4.2
25.5
0.4
4.8
0.2
34.8
0.2
4.5
0.5
0.82
0.1
16.6
0.2
Chamber
Post renewal
of BRH Slurry
343
9.9
22.0
0.4
4.8
0.5
34.6
0.4
4.5
0.5
0.7
0.2
17.8
0.2
Control
Chamber
5.2
0.8
5.6
0.7
0.7
0.5
1.4
0.3
<'
<0.2
<0.2
* Seawater samples were collected 2 hr before renewal of the Black
  Rock Harbor sediment slurry and 3 hr post-renewal of the slurry.
                                    91

-------
ratio is also given for the BRH sediment 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.  The results indicate that BRH sediment samples




are reasonably homogeneous for a given barrel if precautions are taken to




re-mix each barrel before sampling.




      91.  Seawater.  The results for the monitoring of the mussel




exposure system are given in Table 18.  Only the total acid leachable




concentrations present in the unfiltered seawater are reported.




Concentrations for the elements of interest in the control chamber were




generally undetectable by direct injection of seawater samples into the




HGA unit; however, seawater samples from the BRH exposure chamber could




be analyzed by this method*  The results show that the concentrations




for most of the metals are reasonably consistent for the two sets of




samples collected prerenewal and post-renewal of the BRH sediment slurry.




      92.  It is important to know if the concentrations of the various




metals determined in the exposure chamber agree with the measured




particulate concentrations that were delivered by the dosing system.




The total concentration of BRH sediment delivered into the exposure




chamber can be calculated from the Fe concentrations presented in Table




18 assuming Fe is conservative.  Using these data, the calculated




concentrations for BRH sediment are 10.8 and 12.8 mg/L for the prerenewal




samples and postrenewal samples, respectively.  The values actually




measured by filtration of seawater samples from the exposure chamber
                                    92

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                           Table  19

Average Fe/Metal Ratios (± Standard Deviation) for the Exposure
       Chamber Seawater and Black Rock Harbor Sediments
                                               Exposure  Chamber
 Fe/Metal             BRH Sediment                  Seawater

 Fe/Zn                 24.8 + 0.9                  17.7 + 2.4

 Fe/Mn                 80.9 + 7.9                  77   + 17

 Fe/Cu                 12.4+0.5                   9.1 + 1.0

 Fe/Pb                 78.4 + 2.6                  70   +8

 Fe/Cd                 1270 + 45                  420   + 156

 Fe/Cr                 20.8 + 0.98                 18.4 + 1.9
                               93

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during the course of  the exposure period range  from  8.19  to  10.33 mg/L.




The values determined for  the control chamber during  this same period




range from 1.45  to  2.02 mg/L.  The calculated concentration  of BRH




sediments in the exposure  chamber are, therefore, only  10 to  20 percent




higher than the  actual range of concentrations  determined over the entire




time course of the  experiment.  Iron was used to make these  calculations




because Fe has the  highest concentration of any metal measured in BRH




sediment and should be affected less by contamination during  seawater




sample analysis.




      93.  The inter-elemental ratios of the metals  (i.e. Fe/metal) are




given in Table 19.  This table also contains the Fe/metal ratios of the




BRH sediment samples.  Theoretically the Fe/metal ratios determined in




the seawater samples  should agree with the Fe/metal ratios for the BRH




sediment samples.   There are, however, some difficulties with this




concept.  The seawater metal concentrations determined were analyzed at




concentrations 1000 times  lower than were determined in the bulk BRH




sediment samples.   Therefore, detection limits  and contamination of




seawater samples could affect the metal concentrations and their sub-




sequent Fe/metal ratios.   This is important since the measured metal




concentrations are used to determine bioaccumulation factors  of metals




for the exposed mussel samples.  The Fe/metal ratios for the  exposure




chamber seawater samples presented in Table 19  are the average values




for the prerenewal and postrenewal of the BRH slurry.  The Fe/metal




ratios for the BRH sediment samples were calculated from the  data presented




in Table Bl.   The BRH sediment Fe/metal ratios are the average of the




individual ratios for the nine samples.  The Fe/metal ratios  for the
                                    94

-------
seawater samples compare favorably with the BRH sediment ratios for all




elements listed except Cd.  The Fe/Cd ratio is different from the sediment




ratio by a factor of 3.  This indicates that a secondary source of Cd




was present in the exposure tank or that the seawater samples were con-




taminated with Cd during collection.  The detection limit for Cd in




seawater using the present analytical techniques is about 0.1 to 0.3 ug/L.




The concentration of Cd determined in the exposure chambers was 0.8 and




0.7 ug/L for the two sets of samples collected.  Therefore, the concentration




of Cd determined in the exposure chamber was very close to the analytical




detection limit and this probably accounts for the factor of 3 difference




in the ratio compared to the BRH Fe/Cd sediment ratio.  Also, the ratio




of Fe/Zn was corrected for the Zn concentration determined in the control




tank (i.e., 5.4 ug/L).  This Zn concentration is probably due to the




large amount of PVC piping that is used in this facility to carry seawater




from Narragansett Bay to the various laboratory seawater exposure




experiments.  Zinc is used as a catalyst in the production of PVC plastic.




The concentration of Zn for Narragansett Bay at a site near the ERL-N is




about 1 to 2 yg/L.




      94.  The concentration of Hg could not be detected In either the




control chamber or the exposure chamber.  The exposure chamber should




have a Hg concentration of approximately 0.02 ug/L.  This Hg concentration




was calculated by dividing the average Fe concentration in the exposure




chamber seawater samples by the Fe/Hg average ratio for the BRH sediment




samples.   The detection limit for Hg with our present analytical




equipment is 0.05 ug/L.
                                    95

-------
      95.  Arsenic was not determined in the control or exposure chamber




seawater samples.  The As concentration for BRH sediments (provided by




New England Division, Corps of Engineers) is 6.1 mg/kg.  The calculated




ratio of Fe/As for BRH sediments using the average Fe concentration from




Table Bl is 4850.  The theoretical As seawater exposure chamber con-




centration can be calculated by dividing the Fe seawater concentration




with this calculated Fe/As ratio value.  The calculated As concentration




due to the addition of 10 mg/L of BRH sediment to the exposure chamber




is 0.07 ug/L.  The natural concentration of As in seawater is




approximately 1 to 2 ug/L.  Therefore, the natural seawater concentration




of As is about 15 times greater than the BRH sediment As added to the




exposure chamber.




      96. Mussels.  The average metal concentrations and standard




deviations for the mussel samples collected from the BRH exposure chamber




on day 28 are given in Table 7.  All of the inorganic data used to




calculate these averages are given in Tables B2, B3, and B4.  The averages




reported for the control mussel samples in this Table are the averages




for all the control mussel samples and not just day 28.  There is




a statistically significant (a « 0.05) difference between the means for




the Cu concentrations of the control mussel samples collected at the




different times during the experiment.  However, for all the other metals




determined, there is no statistical difference in their means for the




control mussel samples collected during the course of the experiment.




There is no significant difference between the mean Mn, Zn, and As in



the control and exposed mussel samples for the 28-day sampling period




(Student t-test, P<0.05).  However, there is a significant difference
                                    96

-------
between the means for the 28-day control mussel samples and the 28-day




exposed mussel samples for all of the other elements determined (lie.,



Fe, Cr, Cu, Pb, and Cd).  If the average of all of the control mussel




samples (days 0, 28, 56, and 70) are compared to the 28-day exposed



mussel samples, then only the mean Mn concentration for the 28-day BRH



exposed mussels is not significantly different from the mean Mn



concentration for all the control mussel samples.




      97.  During the uptake period (excluding time 0) there is no




significant difference (one way analysis of variance, OC »0.05) of the




means for Fe, Cu, Fb, Cr, and Zn for the BRH exposed mussel samples over



time.  This would indicate that equilibrium was reached for these




metals by the time the first set of exposed mussel samples was collected




(i.e., 1.8 days).  This might suggest that the mussels simply had BRH



sediment in their gut at the time of collection and that the mussels



depurated the BRH sediment at a constant rate during the uptake period*



However, this is refuted by the following.  The Fe, Cr, Cu, Pb,




and Cd concentrations for the mussel samples collected from the exposure



chamber during the first week of depuration (day 35) were still elevated



relative to their mean concentrations in the control mussel samples.



Gut depuration of BRH sediment from the exposed mussel samples should



take place faster than 7 days.  Also, Pb did not depurate readily



from the BRH exposed mussels between day 35 and day 70.



      98.  The bioaccumulation factors (BAFs) for the metals determined




in the 28-day exposed mussels are given in Table 20.  To determine the BAF



values, the following calculations are made: (a) the average metal concen-




trations for the control mussels are subtracted from their respective
                                    97

-------
                          Table  20
     Metal Bioaccumulation Factors  for Mussels  Exposed^jio
                 Black Rock Harbor  Sediment*

Metal
Fe
Zn
Mn
Cu
Pb
Cd
Cr
Mussel
BAF
972
6513
-208
1243
1977
6567
1331
Mussel
28 Day /Control
2.6
1.9
0.9
4.6
2.8
2.7
11.4
* The ratios reported are the day 28 mussel sample  averages
  divided by their respective average control concentrations
                               98

-------
                       Table 21
Average Fe/Metal Ratios (± Standard Deviation) for Control
        Mussels and Black Rock Harbor Sediment
Fe/Metal           BRH Sediments               Mussel Control

Fe/Zn               24.8 + 0.9                    1.1 ± 0.2

Fe/Mn               80.9 + 7.9                   18.2 + 5.A

Fe/Cu               12.4 + 0.5                   17.9 + 4.2

Fe/Pb               78.4 + 2.6                   40.7 + 10.2

Fe/Cd              1270   + 45                    73.8 ± 9.8

Fe/Cr               20.8 + 0.98                 100   + 33
                            99

-------
metal concentrations for day 28 BRH exposed mussels; and  (b) the resultant




metal concentrations are divided by their respective BRH exposure chamber




seawater metal concentrations.  The units of measure for the mussels are




in micrograms per gram dry weight and the seawater concentration units




are in micrograms per milliliter.  The ratios of the average metal




concentrations for the day 28 BRH exposed mussels to the average




metal concentrations for the control mussel samples are also reported




in Table 21.  The ratios are a different method of representing the




metal accumulation in the day 28 BRH-exposed mussels.  The BAF values




tend to give an impression of a large uptake by the mussels for the




metal concentrations determined.  However, the metal ratios give only




the relative metal concentration increases for the BRH-exposed mussels




versus the control mussel samples in this study.  For example, the BAF




values for Zn and Cr are 6513 and 1331, respectively.  However, the day




28/control ratios are 1.9 and 11.4 for Zn and Cr, respectively.  Using




only the BAF values one might conclude that Zn would have a larger




percent increase than Cr in exposed mussels.




      99.  There was no increase for the Mn concentration in the day 28




BRH-exposed mussels compared to the control mussels.  However, the




increase for Cr for these same mussel samples was a factor of 11.  The




increases for the other metal concentrations determined for the day 28




BRH-exposed mussels compared to the control mussels are generally greater




by a factor of 2 to 5.




      100.  The uptake and depuration curves for the metals determined




in the samples are given in Figures 24 to 27.  There are two day 14




sets of mussel samples collected from the BRH exposure chamber (day 0
                                   100

-------
  600
1

3400

-------
 ~ 15
 E
 Q.
 O.
 - 10
 JO
 Q.
            THE STANDARD DEVIATIONS OF THE AVERAGE
            METAL CONCENTRATIONS ARE DEPICTED AS
            A VERTICAL LINE. ALL Pb AND Cd CONCEN-
            TRATIONS ARE IN  G/G DRY WEIGHT.
                                      o EXPOSED
                                      • CONTROL
         —UPTAKE—h- DEPURATION —
               10  20  30 40 50  60  70
                      TIME  (days)
                      a.  Pb
     8
  E
  Q.
  Q.
°-  6
  3  4
                                      o EXPOSED
                                      • CONTROL
         —UPTAKE
                        DEPURATION-
Figure 25,
         0   10 20  30  40  50  60  70

                    TIME  (days)

                   b.  Cd

         Uptake and  depuration in mussels exposed
               to BRH sediment
                       102

-------
EIOO
ex
   50
 E 8
 a
 a.
          THE STANDARD DEVIATIONS OF THE AVERAGE
          METAL CONCENTRATIONS ARE DEPICTED AS
          A VERTICAL LINE. ALL Cu AND As CONCEN-
          TRATIONS ARE IN nG/G DRY WEIGHT.
                                    o EXPOSED
                                    • CONTROL
          0   10  20  30  40 50 60 70
          — UPTAKE-H-DEPURATION-
                     TIME (days)
                    a. Cu
                                    ° EXPOSED
                                    • CONTROL
        — UPTAKE—I—DEPURATION
          I         	1	1	1	i—
          0   10 20  30  40  50  60 70

                     TIME (days)

                    b. As

Figure 26.  Uptake and depuration  in mussels exposed
                to BRH sediment
                      103

-------
  400

9- 300
   200

   100
   40
E
o.
o.
c
 a30
   20
    10
            THE STANDARD DEVIATIONS OF THE AVERAGE
            METAL CONCENTRATIONS ARE DEPICTED AS
            A VERTICAL LINE. ALL Zn AND Mn CONCEN-
            TRATIONS ARE IN pG/G DRY WEIGHT.
                                     o EXPOSED
                                     • CONTROL
          -UPTAKE	1	DEPURATION —
               10  30  30  40  50 60 70
                      TIME  (days)
                   a.  Zn
                                    o EXPOSED
                                    • CONTROL
        —UPTAKE
                        DEPURATION
           0  10  20  30  40 50 60 70

                     TIME (days)

                    b.  Mn

Figure 27.  Uptake and depuration in mussels exposed
                to BRH sediment
                     104

-------
 to 14 and day 14 to 28).  These two sets have been combined to create




 only one average concentration for day 14 of the uptake period for the



 exposed mussel samples.




       101.  To determine if any relationships exist between any of the




 metals determined for the homogenized mussel samples, correlation




 coefficients (r) were calculated for all metals compared to the Fe




 concentration for each sample.  Iron was chosen as the element to




 compare to the other metals for three reasons:  (a) Fe has the highest




 concentration of all the metals determined in BRH sediment; (b) Fe has




 the smallest percent standard deviation of the  means of the control




 mussel samples for the entire experiment;  and (c) Fe should be less




 subject to contamination during analysis compared to any of the other




 metals determined.




       102.  The  number of sample pairs  (i.e., mussel concentrations of X




 and Y) must  be considered in  order  to evaluate  the probability of  the




 correlation  coefficient being significant  due to  random sampling from




 an  uncorrelated  population.   All  the  correlation  coefficients  used




 in  the following  discussion are based on P values  of  0.05.  For  example,




 24  data pairs  require  a correlation coefficient greater  than 0.381  to




 be  at  the  0.05 level of  significance  (Fisher  1985).




       103.  During  the  uptake period  (24 sample pairs) only Mn was not




 significantly  correlated with Fe.  All  the other metals  showed varying




 degrees of correlation.  The  calculated correlation coefficients for  Fe




versus Cr, Pb, As, Zn, Cd, Cu, and Mn are 0.957, 0.827,  0.635, 0.534,
                                   105

-------
0.490, 0.461, and 0.263, respectively.  None of the metals correlated




with Fe for the control mussel samples or for the mussel samples collected




during the depuration phase of the experiment.




      104.  It Is easily seen that the Fe and Cr concentrations In the




mussel samples covary linearily (r-0.957) during the uptake period (see




Figure 24).  A comparison of the Fe/Cr ratio (see Table 21) for the




control samples versus the BRH sediment samples shows that the control




mussels have a ratio five times that of the sediment samples.  The




only other metal determined that has a Fe/metal ratio larger for the




control mussels versus the BRH sediment is Cu, and this ratio difference




is a factor of 2.  The Fe/Cr ratio difference in the controls and BRH




sediment may be an advantage in determining uptake of BRH sediment in




mussels during the field verification portion of this study.  The




relatively low concentration of Cr in the control mussels versus the




BRH sediment makes Cr an ideal choice for a metal tracer of BRH sediment.




      105.  The uptake/depuration plots of Pb, Cd, and Cu in the mussels




from the BRH exposure chamber are given in Figures 25 and 26.  All three




of these metals are elevated during the uptake period compared to the




control mussel samples, but there is no clear uptake pattern for any of




them.  The correlation coefficients of Pb, Cd, and Cu versus Fe during




the uptake period are 0.827, 0.490 and 0.465, respectively.  All three




of these correlation coefficients are significant (P=0.05) for a




population of 24 sample pairs.  The Pb uptake curve also resembles many




of the same features of the Cr and Fe curves during the exposure phase




of the experiment.
                                    106

-------
     106.  The maximum amount of uptake for Cu occurs early (day 3.5)




into the exposure period compared to all the other metal concentrations




determined and then tissue concentrations decline after 21 days




into the exposure.  The depuration of Cu appears to be steady in




that a slow release of Cu occurs over a 3-week period.  The Cu




concentration declines to the average control mussel concentration




on day 56 and then remains constant until the end of the experiment.




    107.  The metals Pb and Cd do not show the same type of depuration




curve.  Neither of these elements shows a steady decline in concentration




during the depuration period.  At day 70 (day 42 of depuration), the




average Cd concentration for the exposed mussels declined to the control




mussel concentration, but the average Fb concentration for the exposed




mussels did not decline to the control mussel concentration.




    108.  The last three metals determined in the mussels, As, Zn, and




Mn, have one common feature.  During the uptake and depuration periods




the concentrations of these metals in the exposed mussels vary around




their respective concentrations in the control mussels.  Several of




the average concentrations for these metals in the exposed mussels are




lower than the average for the controls during the uptake period.  The




uptake and depuration plots for these three metals are given in Figures




26 and 27.  The correlation coefficients for As, Zn, and Mn versus




Fe are 0.635, 0.534, and 0.263, respectively, for the uptake portion of




the exposure period.  Of these three metals, only As and Zn have




significant correlation coefficients (P-0.05) for a population of 24




sample pairs.
                                  107

-------
                                                    30
 I
 O
 Q.
 in
 UJ
 cc
 O

 o
 UJ
 H
 LJ
 Q
                                   TIME


                              a.   BRH sediment
t
UJ
o
a.
cc.
o
UJ
O
                                                      35
    Figure 28,
                  TIME


         b.  Reference sediment


Capillary column  electron capture gas  chromatograms

        of PF-50  (PCB) fraction
                                   108

-------
                                  Worm Test






Organic Contaminants




      109.  PCBs - Sediment.  The electron capture detection (BCD)




chromatograms of extracts from BRH sediments taken from the exposure tank




show a distribution of PCB compounds with from two to ten chlorine atoms




with the predominant peaks containing five and six chlorine atoms (Figure




28).  Tentative identifications of the peaks are shown in Table 5.




Chromatograms from day 0 and day 40 of the exposure sediment show almost




Identical distributions of compounds.




      110. The ECD chromatograms of reference sediment from the reference




tanks show a distribution of PCBs with from two to ten chlorine atoms.




The distribution of PCBs in the reference sediment shows a relatively




greater abundance of higher molecular weight PCBs than is found in the




exposure sediment.  Chromatograms from day 0 and day 40 of the reference




sediment show almost identical distributions of compounds.




      111.  Sediments from the exposure and reference tanks were sampled




throughout the study.  Samples taken on day 0 and day 40 were analyzed




and the results show only small differences in the concentrations of




PCBs over the study period.



      112.  PCBs - Worms.  The polychaete worms Nereis virens on




arrival at ERLN were large but visually appeared to lose weight




during exposure to BRH dredged material.  Worms from the reference




tank did not appear to lose weight over the duration of the experiment.




Observations of worms after removal from the tanks and during the gut




depuration phase (see Methods, paragraph 20) showed that the exposed
                                   109

-------
 organisms  appeared  to  process  only  small  amounts  of  BRH  material  during




 the uptake phase  (some worms from the  exposure  tank  had  no  sediment  in




 their  guts).  When  the exposed worms were placed  in  reference  sediment




 for depuration, they processed only small amounts of sediment.  Reference




 worms, however, were always full  of sediment prior to gut depuration.




       113.  Chromatograms  of worms  from the exposure and depuration




 study  are  shown in  Figures 29  and 30.   The chromatograms from  the day 0




 worms  show three  predominant peaks  (36, 39, and 48)  which represent




 compounds  with structures  that are  resistant to degradation (Zell and




 Ballschmiter  1980).  The distribution  of  these  peaks maximizes  at the




 Cl, PCBs.   Only trace  amounts  of  peaks No. 1 to 13 are evident.




 Following  exposure  for 14  days the  organisms had  accumulated a




 range of PCB compounds from C\2 to  c^10*  However, comparison of  the




 chromatogram from day  14 worms with the chromatogram  from BRH sediments




 shows that earlier eluting compounds appear to  be  preferentially




 accumulated.  A peak distribution similar to that  observed in the day 14




worms is found in day  28 and day  42 organisms,  but by day 56 (28 days of




depuration) peaks 36 and 39 are beginning to predominate.  Reference




worms showed only minor changes in  peak patterns during  the experiment.




      114.  During the exposure to the Black Rock  Harbor sediments, N^




virens accumulated the PCB A-1254 (Figure 31).  It is not known if the




worms reached steady-state during this 28-day uptake.  Some researchers




have found no indication of equilibrium concentrations being approached




during 32 days of  exposure of worms (N. virens) to sandy sediment spiked




with A-1254 (McLeese et al. 1980).  Other researchers found that (a)
                                   110

-------
                     a.   Day  0
I-
UJ
O
                        TIME
                     b.   Day 14
 Figure 29.   Capillary column electron capture gas
 chromatograms of PF-50 (PCB) fraction for worms
              exposed to BRH sediment
                         111

-------
UJ
en


a
c/j
LJ
a

D
; >

' >
  Figure 30,
                      TIME 	>



                   b.  Day 56



Capillary column electron capture  gas  chromatograms of PF-50

   fraction for worms exposed to BRH sediment
                                     112

-------
  1000-
in
CM
CD
u
a.
   100
                                  exposure
                                   tank
                    control

                      tank
     10
                 UPTAKE
               10
                      DEPURATION-
         20      30      40

              TIME  (days)
50
60
   'Figure 31.
Concentration of total PCBs (as A-1254) in worms

 exposed to BRH sediment versus time
                            113

-------
steady-state for N. virens exposed to A-1254  In naturally  contaminated

sediments was reached by day  30 and day  40 depending  upon  the  sediment

(Rubinstein et al.  1983); and  (b) steady-state was  reached between  35

days and  100 days  (Interpolation of graphical data) for  JJ.  diversicolor

exposed to sediments spiked with different levels of  PCBs  (Fowler et al.

1978).

       115.  The bioaccumulation factors  (BAFs) » PCS  (worm dry wt.)
                                                 PCB  (sediment dry wt.)

for A-1254 in the  present study were 0.20 for the exposed  worms and

1.33 for  the reference organisms at day  28.   Other  researchers have

reported  BAF values of approximately 8 to 10  (dry wt.) for  14.  dlverslcolor

exposed to sediments spiked with A-1254  (Fowler et al. 1978),  and

concentration factors of 3.8 and 10.8 for large and small  N. virens

exposed to A-1254  in sandy sediment after 32 days exposure, although

steady-state conditions were not attained (McLeese et al.  1980).  BAFs

ranged from 0.157  to 1.59 for II. virens exposed to sediments naturally

contaminanted with A-1254 (Rubinstein et al.  1983).

       116. The present study found no depuration of A-1254  contaminants

during the 28-day depuration period (Figure 31).  This is in agreement

with findings that there was no obvious excretion of  PCB by N. virens

during 21 days post-exposure (McLeese et al.  1980).   In contrast, other

researchers reported that 14. diversicolor eliminated  PCB during post-

exposure depuration periods (Fowler et al. 1978).

       117. The uptake and depuration of representative individual PCB

peaks  (identified in Table 5) are shown in Figures A14 through A17.  An

examination of these curves suggests that the lower molecular  weight, more

soluble compounds are accumulated more rapidly than the higher molecular
                                   114

-------
weight compounds, but no significant depurations were apparent.




Bioaccumulation data for the individual peaks are shown in Tables 22 and




23.  While the data are not conclusive, there appears to be a slight



increase in BAF in the range of the C15 and Clg PCB peaks.  The reason




for the lower BAF values observed in th6 exposed versus the reference




worms may be the result of the organic matter content of the sediments.




The reference sediment contained 1.8% organic matter while the exposure



sediment contained 5.9%.  Other researchers have suggested that the



organic content of sediments play a key role in the availability of




organic pollutants to benthic organisms, with higher organic content



decreasing the bioavailability of contaminants (Rubinstein et al. 1983).




      118. Another reason for the relatively low exposure BAF values




found in the present study may result from the low feeding rates observed




for the worms during the study period.  Since the organisms fed little,




if at all, and appeared to lose weight during this study, a significant



portion of the accumulation of PCBs that occurred may have resulted



from bioconcentration of contaminants from interstitial water.  This




route of uptake has contributed significantly to the PCB body residues




of Arenicola marina and Vt_. diversicolor in laboratory exposures (Courtney



and Langston 1978).



      119.  PAH - Sediment.  Sediment samples collected from the exposure




and reference tanks during the worm exposure study showed distributions of




PAH compounds which are typical of distributions found in heavily




contaminated and lightly contaminated sediments, respectively.  A much



more detailed analysis of these sediments can be found in Rogerson et al.




(1983).
                                   115

-------
                   Table 22




PCB Bioaccumulation Factors. Exposed Worms Day  28

Peak No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
BAF
.10
.57
.27
.16
.30
.20
.15
.07
.20
.16
.19
.17
.39
.39
.20
.29
.13
.22
.19
.09
.07
.30
.38
.25
.28
.22
.36
.09
.22
.26
Peak No.
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54






BAF
.21
.24
.04
.21
.35
.33
.32
.16
.26
.19
.29
.30
.21
.16
.23
.23
.23
.20
.19
.17
.19
.12
.15
.06






                    116

-------
                             Table 23




PCB Bioaccumulation Factors of Worms in Reference Sediment - Day 28
Peak No.
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
BAF
.70
.96
1.6
.57
.43
.84
2.6
1,5
1.4
.97
1.3
.38
2.7
1.0
1.6
1.5
.57
1.2
2.9
1.8
1.1
2.7
1.6
4.0
1.8
2.3
1.5
.96
1.6
1.3
.90
1.9
1.4
.94
1.5
.85
1.5
1.1
                                117

-------
       120.  PAH - Worms.  During the exposure to BRH dredged material,




the polychaete worm N. virens accumulated PAH contaminants and the




pesticide Ethylan.  Reference worms also contained PAH compounds; however,




the concentrations of PAH compounds in the reference worms following




exposure were similar to pre-exposure concentrations.  Ethylan was not




detected in the reference worms.  The uptake and depuration of the sum




of the PAH compounds identified in Table 11 are shown in Figure  32.




This curve is dominated by the lower molecular weight PAH compounds.




       121.  Figures A21 to A23 show the uptake and depuration of the




individual PAH compounds and the Ethylan during the worm exposure study.




The lower molecular weight compounds (i.e., phenanthrene, anthracene,




fluoranthrene, and pyrene) showed maximum uptakes at day 14, while the




higher molecular weight PAHs and the Ethylan reached their maximum at




day 28.  Due to the limited sampling times and the fluctuating nature of




the data, it cannot be determined if steady-state was reached for the




accumulation of PAH and Ethylan by the worms.




       122. While steady-state may not have been reached for the PAHs and




Ethylan, bioaccumulation factors (concentration in worm (dry weight)/




concentration in sediment (dry weight)) were determined for comparison




purposes.  The data are shown in Table 24.  With the exception of an




unexplained increase in BAF for pyrene in the reference worms, the BAFs




appear to range from almost zero to a few percent.




       123. In the depuration phase the lower molecular weight PAH compounds




appeared to be depurated more rapidly from worms than the higher molecular




weight PAH compounds.  Ethylan was accumulated by the worms from the
                                   118

-------
10,000
 1000-
 a.
 a
 «^

 CO
 X
  100-1
                                EXPOSURE
                                 CONTROL
                UPTAKE
                •4-
      DEPURATION
                                  T
                                          T
              10
         20
30
40
50
—T~

 60
                             TIME (days)
 Figure  32,
Concentration of sum of parent PAHs in worms exposed

      to BRH sediment versus time
                              119

-------
                                 Table 24
              PAH and Ethylan Bioaccumulation Factors - Worms
Compound
Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benz (a )anthracene
Chrysene
Benzo(b)fluoranthene and/or
Benzo(k)fluoranthene
Benzo(e)pyrene
Benzo (a)pyrene
Perylene
Sum of PAHs with MW of 276
Ethylan
Exposed Worms*
.04
.04
.05
.04
.03
.06
.02
.02
.02
.07
.004
.09
Reference Worms**
.14
.008
.04
.28
.004
.01
.005
.01
.005
.01
NDt
NDt
* Calculated using mean (n=2) concentrations of PAH and Ethylan compounds
  in exposed worm samples at day 28 divided by mean (n=3) concentrations
  of compounds in exposure sediments (dry weight).

**Calculated using concentration of PAH and Ethylan compounds in reference
  worms at day 28 (n-1) divided by concentration of compounds in reference
  sediments (n=l)(dry weight.

t ND - not determined in reference worms.
                                  120

-------
exposure sediments and depurated similar to the higher molecular weight




PAHs.




      124,  jetroleum Hydrocarbons - Worms.  The UCM patterns found in




the worms following exposure to BRH sediments, when compared to those




of the sediment, were shifted toward lower molecular weight compounds



(Figure 33).  Only small changes were observed in these patterns during



the depuration phases.  The concentrations of petroleum hydrocarbons in



the exposed worms during the uptake and depuration phases of the




experiment are shown in Figure 34.  The maximum concentration observed



was at day 28.  Concentrations of total petroleum hydrocarbons appeared




to decrease during depuration.  Only small changes in the patterns and




concentrations of total petroleum hydrocarbons in the worms from the




reference tank were observed.



      125.  Comparisons of Bioaccumulation - Mussels and Worms.



The determination of bioaccumulation or bioconcentration factors for



the organic compounds found in the exposed mussels and worms in this




study depends upon whether the accumulation is considered to have come




from the dissolved phase, the particulate phase, or both.  If the mechanism



of accumulation is direct uptake from the aqueous phase, then bio-




concentration factors (BCFs) may be utilized.  These factors are usually



determined in experiments where organisms (usually fish) are exposed to




known concentrations of the compound in the water, and where SPM is



usually not present.  BCFs are determined by dividing the concentration




of contaminant in the organism at steady-state by the dissolved con-



centration in the exposure water.  Since many organic pollutants have




low aqueous solubilities and high lipid solubilities, BCFs are usually
                                   121

-------
t
LJ
O
0.
cn
UJ
cc


§

o
LJ
                                                        TIME
  Figure 33.  Capillary column flame ionization detector gas chromatogram  of PF-50  fraction  from worms

  exposed to BRH sediment for 28 days.  This fraction contains mostly  straight  chain,  branched,  and

                                      cyclic saturated hydrocarbons

-------
500r
  10
   5L
                                       exposure
                                        tank
                          o control
                              tank
UPTAKE
DEPURATION
            10     20     30     40

                           TIME  (days)
                             50
                    60
70
 Figure 34.  Concentration of total petroleum hydrocarbons in worms
                 exposed to sediment versus time
                            123

-------
high.  BCFs calculated utilizing the concentration of  contaminants  in




the mussels (dry wt) at day  28 divided by  the concentration  of  contaminant



in the filtrate from the exposure water  show the expected high  values




(Table 13).  In addition, a  direct relationship between  Log  P and the




Log BCF of the compound is observed, as  has been found in other studies




(Ernst 1977; Geyer et al. 1982).




      126.  With the exposure and possible accumulation  from both dissolved



and particulate phases, bioaccumulation  factors (BAFs) defined  as the




concentration of contaminant in the mussels at steady-state  divided by




the concentration of contaminant in the  unfiltered water are appropriate.




In the present study, these  calculations show Log BAF  for PCBs  and PAHs




which are constant at approximately 4.2  over a range of solubilities or




n-octanol/water partition coefficients (Tables 9 and 15)   (see paragraph




65 for possible explanations for this constancy).




      127.  If the mechanism of accumulation is only direct uptake from



SPM, then BAFs calculated using the concentration of contaminant in the




mussels at day 28 (dry wt) divided by the concentration of contaminant in




the SPM (dry wt) are appropriate.  BAFs  calculated this way are less than



0.3 for PCB compounds.  Greater variability was found  for the PAH calculations




(Table 25).




      128.  One simplified view of bioaccumulation uses the concept of




organisms as lipids and other adsorbing materials in a semi-permeable




membrane.  If the assumptions are made that membrane transport and the




quantity and adsorption efficiences of the adsorbing materials in different



organism types are approximately equal, then exposure  of these  "organisms"




to equivalent exposure environments,  until attainment  of steady-state,
                                   124

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                                 Table  25

           Mussel Bioaccumulation Factors Calculated from Filters

PCBs
Peak No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34


BAP*
.18
.23
.17
.18
.19
.10
.18
.18
.19
.19
.19
.20
.15
.17
.17
.17
.17
.19
.19
.16
.16
.15
.16
.17
.15
.16
.16
.18
.23
.16
.15
.15
.16
.15

PAHs
Compound
Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benzo(a)anthracene
Chrysene
Benzo(b)f luoranthene and/or
Benzo(k)f luoranthene
Benzo(e)pyrene
Benzo(a)pyrene
Perylene
Sum of PAHs with MW of 276






















(continued)

BAF1
.07
.12
.15
.20
.25
.22
.13

.11
.11
.05
.06























* BAFs calculated using mean mussel concentration of day 28 (n-3)/mean
  concentration of compounds on day 28 filter (n«3) assuming .009 g
  BRH sediment (dry weight)/liter.
                                  125

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                             Table  25  (Cont'd)
PCBs

Peak No.          BAF*
  35              .16
  36              .15
  37              .15
  38              .06
  39              .14
  40              .09
  41              .15
  42              .15
  43              .14
  44              .02
  45              .12
  46              .10
  47              .17
  48              .08
                                  126

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                                  Table  26

                  Comparison of BAFs from Mussels and Worms

PCB Peak No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
. 19
20
21
22
23
24
25
26
27
28
29
30
31
32

BAF - Exposed Mussels*
.71
.22
.28
.21
.24
,11
.24
.33
.26
.24
.24
.25
.23
.20
.27
.27
.25
,25
.27
.25
.24
.24
.24
.25
.24
.24
.25
.27
.35
.24
.24
.24
(Continued)
BAF - Exposed Worms**
.10
.57
.27
.16
.30
.20
.15
.07
.20
.16
.19
.17
.39
.39
.20
.29
.13
.22
.19
.09
.07
.30
.38
.25
.28
.22
.36
.09
.22
.26
.21
.24

 * Bioaccumulation factors calculated using mean (n=3) concentrations of
   PCBs, PAHs, and Ethylan in exposed mussels at day 28 divided by whole
   unfiltered water concentration at day 28 (nsO) converted to concentration/
   gram dry wt. BRH SPM using a .009 g (dry wt.)/liter.

** Calculated using mean (n=>3) concentrations of PCBs in exposed worms at day
   28 divided by mean (n»3) exposure sediment concentration (dry wts).
                                     127

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                             Table 26  (Cont'd)
PCS Peak. No.
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
BAF - Exposed Mussels
.26
.22
.24
.22
.22
.07
.22
.14
.22
.25
.23
.02
.18
.16
.22
.12
.09
.02
.05



BAF - Exposed Worms
.04
.21
.35
.33
.32
.16
.26
.19
.29
.30
.21
.16
.23
.23
.23
.20
.19
.17
.19
.12
.15
.06
PAH Compounds          BAF - Exposed Mussels           BAF - Exposed Worms*
Phenanthrene
Anthracene
Fluoranthene
Pyrene
Benzo ( a ) anthracene
Chrysene
Benzo (b)fluoranthene and/or
Benzo (k)fluoranthene
Benzo (e)pyrene
Benzo(a)pyrene
Perylene
Sum of PAHs with MW of 276
Ethyl an
.04
.06
.18
.22
.40
.37
.22

.18
.16
.09
.08
.33
.04
.04
.05
.04
.03
.06
.02

.02
.02
.007
.004
.09
* Calculated using mean (n=2) concentration of PAH and Ethylan compounds
  in exposed worm samples at day 28 divided by mean (n=3) concentrations
  of compounds in exposed sediments (dry wts).
                                   128

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should result In equivalent "organism" concentrations.  In the present

study, the exposure environments for the mussels and worms included

contaminants in both dissolved and particle bound form.  For worms, the

contaminants in the sediment pore water represent the dissolved phase;

the particle bound phase is the sediment concentration minus the pore

water concentration.  Since the extraction of sediment utilized in this

study included the contaminants in the pore water with the particle

bound contaminants, BAFs may be calculated utilizing the measured sediment

concentration as a representation of a total exposure concentration.*

In a similar way the total exposure concentration for the mussels is

represented by the unfiltered water samples.  For comparison purposes

the total content of contaminants for both worm and mussel exposures is

assumed to reside on the particles.  BAFs calculated as concentration in

organism (dry wt) divided by the total exposure concentration (dry wt)

of sediment (worms) or SPM (mussels) are shown in Table 26.  The close

correspondence of BAFs for PCBs for both mussels and worms suggests that

bioaccumulation of relatively unreactive FCB molecules may be modeled as

a partitioning of these contaminants between the organisms and either

the sediment or SFM.  Whether the actual bioaccumulation process occurs

through direct transfer of hydrophobic organics from sediment to organism

through the lining of the gut or through a dissolved intermediate phase

is unknown.

      129.  The correspondence between measured and estimated BCF values

found in the mussel studies (paragraph 64) is suggestive of a dissolved


* Not including the concentration of contaminants in overlying water
  which are thought to be very small.
                                   129

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phase intermediate between the contaminants on the SPM and those in the




mussels.  However, the increase in measured BCF with increasing Log P




may result in another way.  The concentrations of PCB compounds in the




aqueous phase decrease by orders of magnitude over the range of PCB




compounds.  Since organisms may accumulate particle-bound contaminants



to constant concentrations (I.e., gut transfer), the division of




organism concentrations by measured aqueous concentrations will give




spreads of BCF values covering several orders of magnitude with BCFs




increasing with decreasing compound solubility (increasing Log P).



Similar relationships are possible in the pore water of the sediments




and in the worms.



      130.  For the more reactive PAH compounds greater differences




were observed in the BAFs for the worms and mussels.  The worms showed



smaller BAFs than the mussels (Table 26).  The reason(s) for these



differences are unclear, but may reflect metabolic differences of the




organisms.



      131.  For modeling bioaccumulation, researchers have suggested




utilizing normalization of sediment concentration to the organic




carbon content of the sediment (site of most of adsorption of hydrophobic




organic compounds) (Karickoff et al.  1979) and normalization of organism




concentrations to the lipid content of the organism (most important site




of storage of organic pollutants).  These concentrations are utilized in




thermodynamic arguments with fugacity concepts and will be the subject



of future publications.
                                   130

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Inorganic Contaminants—Worms




      132.  The average concentrations and  standard deviations




for the day 28 worm samples collected from  the BRH sediment exposure




chamber are given in Table 27.  All of the  inorganic data used to




calculate these averages are given in Table B5.  Data for the time zero




worm samples are not used in the following  discussion since much of the



Fe, Cr, and Cu data are higher than all of  the other samples collected.




The large concentrations of Fe, Cu, and Cr  in the time zero samples are




probably due to the fact that the worms were not allowed to acclimate in




the reference sediment prior to the start of the experiment so that a




firm baseline could have been established.  The time zero worms probably




represent the sediment of the Maine coastline from which they were




collected since these worms had never been  in BRH or REF sediment.




Several data points were eliminated from the uptake, depuration, and



reference worm samples.  These results were rejected by application of




the "Q" test for rejection of an experimental observation (Dean and




Dixon 1951).  Two Cd results were discarded: one from the day 28 uptake




samples and one from the day 40 reference sample.  Also, one value for




Cr was rejected from the day 56 depuration phase of the experiment.




These data points are marked with an asterisk in Table B5.




      133.  The means for the two control mussel Fe concentration are




significantly different (Student t-test, P»0.05) for the two collection




times.  However, there are no significant differences in the control worm




mean concentrations for Cu, Cr, Cd, and Zn for the two collection times.



      134.  The Fe uptake and depuration plot for the worms is given in




Figure 35.  Only the average and standard deviation of the average of
                                   131

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                             Table 27

           Average Trace Metal Concentration for Worms
          Collected from the Exposure Chamber on Day  28*
Metal
Fe
Zn
Cu
Cd
Cr
Worm
28 day
404 + 68
139 + 15
31.3 + 9.9
0.73 + 0.10
5.8 + 2.4
Worm
Control
316 + 75
109 + 16
12.2 + 1.2
0.60 + 0.08
2.3 + 0.6
The control concentrations reported for the worms are the average
of all the control samples and not just day 28.  All concentrations
are in ug/g dry weight.  The standard deviations of the means are also
reported.
                               132

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            THE STANDARD DEVIATIONS OF THE AVERAGE

            METAL CONCENTRATIONS ARE DEPICTED AS

            VERTICAL LINES ASSOCIATED WITH EACH
            DATA POINT
1400
o.
o.

4>
u.
  200
                                       ° EXPOSED

                                       • REFERENCE
 O.
 O.
         — UPTAKE-
 DEPURATION
            10   20    30   40   50   60

                      TIME (days)
                         Fe
                                     o EXPOSED

                                     • REFERENCE
           UPTAKE-
DEPURATION
           10   20   30   40   50   60

                     TIME  (days)
                      b.  Cr
 Figure 35.  Uptake and depuration in worms exposed

                  to BRH sediment
                      133

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each set of exposed and control worm samples are shown in this figure.




There is no significant difference in the Fe concentration of the




day 28 BRH-exposed worms and the day 28 reference sediment-exposed




control worms.  There is no significant difference in the Fe




concentrations for any of the worm samples collected from the BRH exposure




chamber during uptake or depuration (one way analysis variance °^ = 0.05).




      135.  The Cr uptake and depuration plot for the worms is given in




Figure 35.  There is a large standard deviation of the concentration




of Cr for the day 28 BRH-exposed worms compared to the standard deviation




of the Cr concentration for the day 28 reference sediment worms.




However, there is a significant difference (Student t-test, P - 0.05)




between these two concentrations.  Also, the difference between the




concentrations is significant for all the worm samples collected during




uptake and depuration.




      136.  The Cu uptake and depuration plot for the worms is given in




Figure 36.  Like the Cr data the Cu data have a large standard deviation




of the mean for the day 28 BRH-exposed worms.  Also, like Cr, the worm




Cu concentration means for the uptake portion of the study from the BRH




exposure chamber are significantly different from the worm Cu




concentration means for the depuration portion of the study.  The two




control worm sample means for Cu are not significantly different from




each other.  The Cu concentration in BRH-exposed worms declines to the




control worm concentration for the day 42 samples (14 days of depuration).




      137.  The Zn uptake and depuration plot for the worms is given in




Figure 36.  There is no significant difference between the Zn concentrations




for the day 28 BRH exposed worms and the day 28 reference worms.  There
                                   134

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             THE STANDARD DEVIATIONS OF THE AVERAGE
             METAL CONCENTRATIONS ARE DEPICTED AS
             VERTICAL LINES ASSOCIATED WITH EACH
             DATA POINT          	
     40

    a 30
    a
   <3 20

      10
                                    o EXPOSED
                                    • REFERENCE
              UPTAKE-
                 •DEPURATION-
             10   20   30   40   50
                     TIME (days)
                     a.  Cu
                               60
      140
   _  120
   lioo
   * 80
   N  60
      40
      20
                                      o EXPOSED
                                      • REFERENCE
          •	UPTAKE—!	DEPURATION-
              10   20   30  40   50   60
                       TIME (days)

                     b.  Zn
     E
     a.
     a
    •o
    O
.0
      0.5
                                      o EXPOSED
                                      • REFERENCE
               UPTAKE-
                   • DEPURATION-
              10   20   30   40  50  60
                       TIME (days)
                      c.
                          Cd
Figure  36.   Uptake and  depuration  in worms
           exposed to  BRH sediment
                      135

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                               Table 28

               Metal Bioaccumulation Factors for Worms

Metal
Fe
Zn
Cu
Cd
Cr
Exposed to Black Rock Harbor

Worm
BAF
0.0030
0.025
0.0080
0.0058
0.0025
Sediment*

Worm
28 day/control
1.3
1.3
2.6
1.2
2.5
The ratios reported are the 28 day worm sample average concentration
divided by their average controls, respectively.
                                 136

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is also no significant difference between any of the Zn  concentrations


for any of the samples collected from the BRH exposure chamber or the

              b
reference sediment control chamber.


      138.  The Cd uptake and depuration plot for the worms is given in


Figure 36.  There is no significant difference between the Cd concentration


for the day 28 BRH-exposed worms and the day 28 reference sediment


control worms.  The two Cd concentrations for the control samples are


also not significantly different.


      139.    The BAF values for the metals determined in the day 28


BRH-exposed worms are given in Table 28.  These BAF values were calculated


for all the metals even though the Zn, Cd, and Fe data show no significant


difference between the means of these elements for the BRH-exposed and the


reference-exposed control worms.  The BAF values were calculated as follows:


(a)'the average control worm metal concentration was subtracted from


the average day 28 worm concentration; and (b) the corrected concentrations


were then divided by the concentration of the metals determined in BRH


sediment.  The ratios of the average metal concentrations for the day


28 BRH-exposed worms to the average metal concentrations for the control


worm samples are also reported in Table 28.  These ratios are probably


a better representation for the metal accumulation in the day 28 BRH-


exposed worms.  The ratios for Cu and Cr for the day 28  exposed worms


and the reference sediment control worms are 2.6 and 2.5, respectively.


These increases in Cu and Cr, while not large, are significant.  The


calculated ratios for Fe, Zn, and Cd are small and are not significant.
                                   137

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                             PART IV: SUMMARY




                      Mussel Bioaccumulation Study




Organics



      140.  The system utilized to dose mussels with suspensions of




BRH dredged material worked well.  The dose of PCB contaminants monitored




in the exposure tank was quite constant over the study period, and while




the concentration of total PAH compounds was found to show more variability,




this variability appeared to be in total levels of PAH compounds rather




than on a compound-to-compound basis.  It seemed likely that the PAH




variability may have resulted from the unhomogeneous distribution of




soot particles (containing high concentrations of PAH compounds) in the




BRH sediment.  The concentrations of PAH and PCB contaminants in the




control tanks were orders of magnitude below those in the exposure tanks.




      141.  Separation of dissolved and particle-bound PCB and PAH




contaminants resulted in distributions which were logically consistent




with the solubilities of the compounds.  The more water-soluble compounds




were found in the dissolved form while the less water-soluble compounds




were found associated with the particles.




      142.  Based on comparisons of measured and estimated Kps for the PCB




and PAH contaminants in the exposure tanks, it appeared that  equilibrium




conditions were not reached in the residence time of the  suspensions  in




the dosing system.




      143. During the first 7 days of  exposure, mussels in the exposure




tank showed a rapid uptake of PAH and  PCB  compounds.  There was an un-



explained  decrease in the  concentration in mussels during the next  2  weeks




for most  compounds, and the highest  concentrations were found at day  28.
                                    138

-------
      144.  PCB compounds with molecular weights above Cly PCB were not




effectively accumulated by the mussels, and, similarly, PAHs of higher




molecular weight were not accumulated as much as some lower molecular




weight compounds.




      145.  Application of a non-linear model to the mussel data indicated




that steady-state concentrations of PCBs had been reached during the 28-




day exposure period.  From the shapes of the uptake curves for the PAH




compounds, it appeared that steady-state concentrations of PAHs also had




been reached during the 28-day exposure.




      146.  While the dominant method of accumulation of PAHs and PCBs




(i.e., uptake from water or uptake from the particles) could not be




determined from these studies, measured BCFs (assuming uptake from aqueous




phase only) showed increasing BCFs with increasing Log n-octanol/water




partition coefficients (decreasing aqueous solubilities) and reasonable




agreement with BCFs estimated from a correlation (BCF vs. Log P) in the



literature.  In contrast, bioaccumulation factors (BAFs; calculated using




unfiltered water concentrations) for PCBs and PAHs were relatively




constant over the range of PAH and PCB contaminants examined.  The




constancy of these BAFs suggests that similar processes determined the




distributions of these compounds.  In this regard, bioaccumulation in the




exposure tanks may be viewed as the result of two processes competing for




the dissolved phase contaminants:, readsorption by the SPM, and bio-




concentration of dissolved contaminants by the mussels.  Alternatively,




the constant bioaccumulations observed may have resulted from similar




constant processes like direct transfer of contaminants through the gut.
                                   139

-------
       147. In general, depuration rate in mussels appeared to be Inversely




related to Log P for PCB compounds; however, some higher molecular weight




PCB compounds were lost at higher rates than lower molecular weight




PCBs.  Those compounds with recalcitrant structures appeared to be




depurated most slowly.  The depuration of PAH compounds also appeared to




be inversely related to Log P.  The concentrations of petroleum hydrocarbons



measured in the mussels over the exposure and depuration period generally




followed the patterns observed for the other organic contaminants.




       148.  Control mussels, which contained only low concentrations of




contaminants, showed only small fluctuations in concentrations of organic




contaminants during this study.




Inorganics




       149.  The acid-soluble trace metal concentrations in the seawater




were generally consistent with the quantity of BRH sediment added to the



exposure chamber.  The total concentration of BRH sediment added to the




seawater could be calculated from the Fe concentration determined in the




seawater exposure chamber.  The interelemental ratios determined in the




exposure chamber also compared favorably with the interelemental ratios




determined for BRH sediments.




       150.  Statistically, there was no difference for the respective means




of Fe, Cr, Mn, Pb, Cd, Zn, and As concentrations over time for the control




mussels collected from the control chamber.  There was, however, a significant




difference in the mean Cu concentrations for the control mussel samples




collected over time from the control chamber.  The means of Fe, Cu, Cr,



Zn, Pb, Cd, and As for the day 28 BRH-exposed mussels were significantly




different from their respective means for the control mussels.
                                   140

-------
      151.  Typically, metal BAFs over  1000 were calculated for the day




28 mussels collected from the BRH exposure chamber.  A different




impression of the uptake is observed in the ratios of metal concentrations




in day 28 exposed mussels divided by the metal concentrations in the




control mussels.  The ratios which indicate only the relative metal




concentration increases for BRH-exposed mussels versus the control mussel




samples ranged from 0.9 to 11.




      152.  The uptake patterns of Fe and Cr for the exposed mussels were




almost identical.  The correlation coefficient for Fe versus Cr in the




BRH-exposed mussels was 0.957.  The relatively low concentration of Cr



in the control mussels versus that in the BRH sediment makes Cr an




ideal choice for a metal tracer of BRH sediment.  The mussel con-




centrations of Fe, Cr, Cu, and Pb were all elevated during the uptake




period compared to the control mussel samples; however, only Pb and Cr




correlated well with Fe in the mussels during the uptake period.  The




concentrations of Zn, As, and Mn in the exposed mussels varied around




their respective concentrations in the control mussels.




      153.  The depuration patterns of Fe and Cr from the mussels




exposed to BRH sediment appeared to be identical.  Both elements declined



to control concentrations after 2 weeks of depuration.  The depuration




of Cu appeared to begin before the end of the exposure period.  The Cu




concentration in the BRH-exposed mussels fell to the control mussel




concentrations after 3 weeks of depuration.  Neither Pb or Cd showed




a steady decline in concentration in the exposed mussels during the



depuration period.  The concentrations of Mn and As in the exposed




mussels were generally below the concentrations of Mn and As concentrations
                                   141

-------
of the control mussels during depuration.  The average Zn  concentrations




in the exposed mussels were elevated compared to the average control




Zn concentration during the depuration period.  However, the standard




deviations of the average concentrations for Zn in the exposed mussels




overlapped with those for the control mussels during the depuration




period.






                        Worm Bioaccumulation Study




Organics




      154.  The exposure of worms Nereis virens to BRH sediment resulted




in accumulation of organic compounds even though the organisms showed




little evidence of feeding during the experiment.  The PCBs accumulated




by the worms showed a pattern which was similar to the pattern observed




in the sediment.  The PCB contaminants accumulated showed no apparent




decreases in total concentrations over the depuration period.  Exposed




worms accumulated PAHs to concentrations which were orders of magnitude




greater than those in the reference worms.  In contrast to the PCBs,




which showed no concentration decreases during the depuration period,




the concentrations of PAHs in the worms fell rapidly during depuration.




      155.  The petroleum hydrocarbons found in worms (measured as an




unresolved complex mixture) increased during the uptake phase and




decreased during the depuration phase.




      156.  Reference worms showed relatively constant low levels of




organic pollutants over the exposure and depuration study.




      157.  The bloaccumulation factors observed for the PCBs in the




exposed worms were lower than those found for worms in the reference




sediment.  This may have resulted from the apparent poor health of the
                                   142

-------
exposed organisms or from decreased bioavailability of PCB contaminants




in sediments with high organic carbon (i.e., Black Rock Harbor sediments).




Alternatively, the lower bioaccumulation factors found for worms in




Black Rock Harbor sediment may result from steady-state values not




being attained during the 28-day exposu're period.  BAFs for PAH compounds




in the worms were lower than BAFs for the PCBs, and no consistent




differences between exposed and reference BAFs were found for PAHs.  The




differences observed in the BAFs for PCBs and PAH may reflect metabolic




or bioavailability differences, or may result from steady-state values




not being attained during the exposure period.






Inorganics




      158.  There was no significant difference between the mean Fe, Zn,




and Cd concentrations in the day 28 BRH-exposed worms compared to the




reference sediment exposed worms.  There was a significant uptake of Cu




and Cr for the day 28 BRH-exposed worms.  The calculated ratio of the mean




Cu and Cr concentrations for the day 28 BRH-exposed mussels and the




reference sediment exposed mussel were 2.6 and 2.5, respectively.  The




depuration of Cu and Cr was complete 2 weeks after the exposure period.







                 Bioaccumulation Mussels and Worms—Organic^







      159.  Exposure to organic contaminants in dissolved form and on SPM




(mussels) and in pore water and sediments (worms) can be simplified by




assuming that the total exposure concentration of PCBs resides on the



particles or the sediments.  When this is done the calculated BAFs for




both mussels and worms are quite similar.  Similar calculations with
                                   143

-------
the more reactive and possibly less available (due to incorporation in




soot particles) PAHs show greater differences.  Similarities in worm




and mussel BAFs suggest that modeling bioaccumulation of some organics




(at least less reactive compounds like PCBs) as a partitioning of




contaminants between the organisms (worms and mussels) and the sediment




or suspended sediment shows promise as a predictive technique for



assessing the accumulation of organic contaminants from dredged material




and other mixed wastes.
                                   144

-------
                           PART  V:  RECOMMENDATIONS

                                 Mussels


       160.  Due to  the apparent discrepancy between estimated and

measured partition  coefficients for organic compounds, it is recommended

that partitioning studies  be conducted to determine the time to equilibrium

and the soluble/particulate distributions of organic and inorganic

compounds under both aerobic and anaerobic conditions.

       161.  Due to  the variability in concentrations of organics and

inorganics observed in mussels during the uptake phase, the following

studies are recommended:

        _a.  Studies should be conducted to broaden our understanding
            of the nutritional requirements of these organisms and
            the potential nutritional value of the sediments.

        _b.  Future studies should be conducted to examine the
            contribution of contaminants on sediment in the gut of the
            exposed mussels to the concentrations found in extracts of
            whole organisms.

       162.  It is recommended that a range of concentrations be used

in mussel exposures to establish the constancy of bioaccumulation

factors at different exposure concentrations.


                                  Worms


       163.  Due to the poor feeding behavior of the worms in this

exposure study, it is recommended that further studies be conducted

to ensure that worms remain healthy and have adequate nutrition during

exposure and depuration studies with sediments which are heavily contamin-

ated with organic and inorganic compounds.  This research should consider

the utilization of standardized control sediment in exposure studies.
                                   145

-------
These standard sediments could be used to dilute toxic sediments and

to serve as carriers for the nutritional needs of the organisms.

      164.  It is recommended that worms be held in reference sediment

prior to initiation of exposure studies for a time sufficient to allow

worms to adjust their contaminant levels to those of the reference

sediment.

      165.  As adequate exposure conditions (i.e., no adverse effects)

become available, it is recommended  that longer term bioaccumulation

studies be conducted to ensure that  steady-state levels are  reached

for both the organic and inorganic contaminants in these organisms.

                                 General


       166.   In order to determine the  potential for bioaccumulation of

sediment-bound contaminants,  it  is  recommended that research be  undertaken

to develop a short-term abiotic  test to enable prediction  of the bio-

availability and  bioaccumulation of  contaminants.

       167.   In order to link  the laboratory bioaccumulations with

potential bioaccumulations in the  field at  the disposal  site for BRH

dredged  material  (under  the Field  Verification Program),  it  is  recom-

mended  that:

  £.     Field  bioaccumulation samples  be  analyzed  for  the  same  contaminants
         that were accumulated in laboratory studies.

  b_.     Where  possible,  the same organisms  (or  similar surrogate organisms)
         be  used  in field  and  laboratory  studies.

  £.     If  the same organisms cannot be  deployed or are not indigenous
  ~~     at  the disposal site, studies  be  undertaken to compare
         bioaccumulation between indigenous  and surrogate organisms.

  d.     Exposure concentrations at the disposal  and reference site should
  ~     be determined at least seasonally to enable estimation of  mean
         annual exposure levels in water and sediments.
                                    146

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