United States
             Environmental Protection
             Agency
            Great Lakes National
            Program Office
            536 South Clark Street
            Chicago, Illinois 60605
EPA-905/3-84-007
November 1984
v>EPA
Flow-through  Bioassay
For Measuring
Bioaccumulation of
Toxic Substances From
Sediment
                 Do not WEED. This document
                 should be retained in the EPA
                 Region 5 Library Collectioii.

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                                                         EPA-905/3-84-007
                                                         November 1984
                    Flow-through  Bloassay For Measuring
             Bloaccumulatlon  of Toxic  Substances  From Sediment
                              Michael J. Mac
                              Carol  C. Edsall
                            Robert J. Hesselberg
                           Richard E. Sayers, Jr.

                       U.S.  Fish and Wildlife Service
                       Great Lakes Fishery Laboratory
                        Ann Arbor,  Michigan 48105
                               Final  Report
                                 May  1984
                   Interagency Agreement DW 930095-01-0

                             Project  Officer
                            Anthony Klzlauskas
                          Remedial Program Staff
                   U.S. Environmental Protection Agency
                   GREAT LAKES NATIONAL PROGRAM OFFICE
                   U.S. ENVIRONMENTAL PROTECTION AGENCY
                     536 SOUTH CLARK STREET, ROOM 958
                         CHICAGO, ILLINOIS 60605
^/Contribution No. 616 of the Great Lakes Fishery Laboratory

                                 U.S. Environmental Protection Agency
                                               Sowlwartf, 12m Ffosf
                                 Chicago.lt  60604-3590

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                               DISCLAIMER

     This report has been reviewed by the Great Lakes National Program
Office, U.S. Environmental Protection Agency, and approved for publication.
Approval does not signify that the contents necessarily reflect the views
and policies of the U.S. Environmental Protection Agency, nor does mention of
trade names or commercial products constitute endorsement or recommendation
for use.
                                J!

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                                  FOREWORD


The Great Lakes National Program Office (GLNPO) of the United States Enviro-
mental Protection Agency was established in Region V, Chicago, to focus
attention on the significant and complex natural resource represented by the
Great Lakes.

GLNPO implements a multi-media environmental management program drawing on
a wide range of expertise represented by universities, private firms. State,
Federal, and Canadian governmental agencies, and the International Joint
Commission.  The goal of the GLNPO program is to develop programs, practices
and technology necessary for a better understanding of the Great Lakes Basin
ecosystem and to eliminate or reduce to the maximum extent practicable the
discharge of pollutants into the Great Lakes system.  GLNPO also coordinates
U.S. actions in fulfillment of the Great Lakes Water Quality Agreement of
1978 between Canada and the United States of America.
                                     iii

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                                 CONTENTS








INTRODUCTION 	   1



MATERIALS AND METHODS 	   2



RESULTS 	   8



DISCUSSION 	  11



REFERENCES 	  15
                                     IV

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                                  FIGURES

                                                                        PAGE

1.  D1 agram of Sediment Exposure System	3
                                  TABLES
1.  Physical  and Chemical  Characteristics from the Three Collections.
    Dates of Collection and Dates of Various Tests are also Presented	6
2.  Summary of Test Conditions and Mortality of Organisms	9
3.  Mean Weight, Lipid Content, and PCB and Zn Concentrations  in
    Analyzed Samples	10

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                             ACKNOWLEDGEMENT
The authors acknowledge the technical assistance of David Nortrup,
Susan Praasterink, and Dr. James G. Seelye.  We also acknowledge
personnel at the U.S. Fish and Wildlife Service, Hammond Bay
Biological Station for assistance with collection of ollgochaetes.
This work was supported In part by the U.S. Environmental Protection
Agency, Great Lakes National  Program Office under Interagency Agree-
ment DW 930095-01-0 with the U.S. F1sh and Wildlife Service, Great
Lakes Fishery Laboratory.
                                    v1

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                                 Introduction

     Over 10 million cubic meters of  sediment are dredged annually from Great
Lakes waterways.   Because much  of this material is taken from harbors,
connecting channels, and other  nearshore areas that often are contaminated
with toxic substances,  the sediments  proposed for dredging need to be
evaluated for the presence of bioavailable contaminants and the potential  for
toxicity to the biota.   Sound decisions on the appropriate disposal of the
dredged material  can be made only after such an evaluation.  Presently, no
standardized procedure  exists for evaluating dredged material in freshwater
systems although criteria for discharge of dredged material into marine waters
have been developed  (USEPA/CE 1977).  In the ocean discharge guidelines, it is
recommended that bioassays be conducted on liquid, solid, and suspended
particulate phases of dredged material.  Because it appears that the solid
phase has the greatest  potential  for  environmental damage and because
measuranents of bioaccumulation must  be made to evaluate sediments for
disposal (USEPA/CE 1977, Seelye and Mac 1983), we developed a bioassay for
testing the solid phase of dredged material that measures the survival of
organisms and, perhaps  more important, the bioaccumulation of toxic
substances.  Although other workers ^have demonstrated the bioaccumulation  of
toxic substances by  aquatic organisms from naturally contaminated sediments
(Peddicord et al . 1980; Rubinstein et al . 1980, 1983; Seelye et al. 1982),
several have used testing  methods that result in unacceptable mortality to
control organisms (Bahnick et al. 1981, Prater et al. 1983).

     Our bioassay is intended to  estimate the potential for bioaccumulation of
contaminants from sediments that  are  not acutely toxic to test organisms,  but

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are suspected of containing  persistent contaminants.  By using test organisms
that are not highly susceptible to toxic compounds, the bioaccumulation test
allows estimation of the potential food-chain accumulation of contaminants
that may occur in local  biota  from surficial sediments.  In practice,
bioaccumulation observed in  this  bioassay by organisms exposed to test
sediments (sediments to  be dredged) would be compared to bioaccumulation
observed from sediments  collected from a reference site (e.g. a disposal site
or open lake), and also  from control sediments (relatively clean sediment).
Decisions could then be  based  on  a comparison of  results between test and
reference sediments to determine  if disposal would cause degradation to the
habitat, and between reference and control sediment to determine if even the
reference material is seriously contaminated.  Although the test is not
intended to be a toxicity test per se, use of test, reference, and control
sediments enables interpretation  of any mortality of organisms that may occur
during the bioassays. High  mortality in bioassays with test or reference
sediment would indicate  acute  toxicity of sediments in the project area.
However if high mortality occurs  in all three sediments, it can be assumed
that the organisms were  not  in a  healthy state at tne time of testing.
     We describe the results of 10-day sediment bioassays in which both
mortality and bioaccumulation  were measured in four aquatic organisms.  We
exposed two infaunal  organisms and two species of fish to test and control
sediments in the laboratory.

                            Materials and Methods

     Sediment bioassays  were conducted in a flow-through system consisting of
eight 39-L glass tanks (Fig. 1).  Each tank received 100 mL/min of 20°C water,

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       RAW WELL WATER
       DEIONIZED WATER
u>
CHILLER
HEATER
                                             HEAD  TANK
                                                                                    THERMO-
                                                                                    REGULATOR
                                                                                             .VALVE


                                                                                              WATER
                                                                                              LEVEL
                                                                                             -STANDPIPE
                                                                                              SEDIMENT
       Figure 1.  Diagram of sediment exposure system.

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softened to a hardness of  about 120 mg/L (as CaCOs) by mixing deionized  well
water with processed  well  water (hardness 442 mg/L, Seelye et al. 1982).
Prior to the start of a test, about 11 kg (5 cm depth) of sediment was added
to each tank; four tanks received contaminated (test) sediment and four
received clean (control) sediment.  Water was then added to the tank and the
sediment was allowed  to settle for 24 hours before test organisms were added.
Tests lasted 12 days.  Organisms were exposed to sediment for 10 days and then
moved to identical tanks containing only flowing water for 2 days to allow for
clearance of ingested sediment from the gut.  During the tests we monitored
water temperature, flow rate, hardness, dissolved oxygen, and sediment redox
potential.  Suspended solids were measured only in the exposures involving
fish.
     We tested two species of fish (fathead minnows, Pimephales promelas, and
yellow perch, Perca flavescens) and two species of invertebrates (an
oligochaete worm, Octolasion tyrtaeum, and the Asiatic clam, Corbicula
fluminea).  Adult minnows  (2-3g) and juvenile perch  ( 2g) were obtained from
the National Fishery  Research Laboratory, La Crosse, Wisconsin; the worms were
collected from the Black River near Onaway, Michigan; and the clams were
obtained from the Sacramento River delta in California.  Test organisms were
held at the Great Lakes Fishery  Laboratory  for at  least 2 weeks and acclimated
to softened water for at least 5 days  before testing.  During this time, fish
were fed Silver Cup!/ pellets, clams  were fed algae  (Chlorella), and
oligochaetes were maintained  in  forest duff in which organic matter was
available for  food.  Organisms  were not  fed during the tests.
i/Reference to trade names does not imply U.S.  Government  endorsement

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     During testing,  we  also  examined the suitability of two other
invertebrates:   Chironomus  larvae and Hexagenia limbata.  Chironomus larvae
are easily cultured and  an  important food chain member in the Great Lakes.   We
rejected them as a test  organism, however, because their small size would
require large numbers of similarly aged individuals to obtain enough tissue for
contaminant analysis. Hexagenia limbata is another important food item in  the
Great Lakes.  We observed high mortality during both holding and testing of
this species.  High mortality of IH. limbata has occurred in other published
sediment studies (Bahnick et  al . 1981).  Because of the small size of
Chironomus and the high  mortality of J4. limbata, no further tests of these
organisms were attempted.
     Either 10 fish,  14  oligochaete worms, or 30 clams were placed into each
tank at the start of  the bioassay.  We netted fish and collected worms and
clams by hand following  10  days of exposure to sediments.  At the end of a
test, we froze all live  organisms whole for contaminant analysis.  In
addition, a sample of test  organisms was frozen prior to the start of the test
for determination of  preexposure contaminant concentrations.  All organisms
were thawed and the clams shelled before homogenization and analysis.
     Control and test sediments were collected just before each test to
minimize chemical changes in  the sediment caused by storage (Table 1).  Three
sets of test sediments were collected with a ponar dredge from the Raisin
River near Monroe, Michigan (41° 54' 1" N, 83° 21' 18" W), and three sets of
control sediments were shoveled from Meadowood Pond in Saline Township,
Michigan (42° 7' 44"  N,  83° 47' 45" W).  In the first exposure, two sets of
tanks were used with  fathead  minnows in one set and oligochaetes in the other.
Yellow perch were tested in sediments from the second collection and clams

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Table 1.  Physical  and chemical  characteristics  of  sediments from the three collections.  Dates of collection and
     dates of various tests are  also presented.
Physical composition (% dry wt.)
Exposures and dates
of test

1.




2.



3.




Fathead minnows
and oligochaetes
(10/20-11/2)
Control
Test
Yellow perch
(11/16-11/28)
Control
Test
Asiatic clams
(1/28-2/9)
Control
Test
Date
Collected




10/12
9/30


11/9
10/25


1/13
1/14
Sand




65
11


51
67


16
36
Silt




19
43


24
16


41
33
Clay




13
35


22
11


32
22
Volatile
solids




2
8


3
4


10
7
Contaminants
PCBs




0.016
31.72


0.013
12.78


0.014
19.60
(ug/g dry wt.)
Zn




34
244


31
147


67
162

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were tested In sediments from the third  collection.  Subsamples of sediment
were taken before placement in test tanks  and  frozen for later chemical and
physical analysis.

     Sediments and test organisms were analyzed  for PCBs and In to examine the
bioaccumulation potential  of both an organic and an inorganic contaminant
directly from contaminated sediment.  We analyzed PCBs in sediment and in test
organisms by gas chromatography (GC), using methods previously described by
Seelye et al. (1982).  Samples of tissue and sediment for Zn analysis were
digested in HN03 according to the method of May and Brumbaugh (1982) except
that no perchloric acid was used, and Zn concentrations were determined on a
Perkin Elmer Model 228 atomic absorption spectrophotoraeter equipped with an
HGA2200 graphite furnace.   Absorption values for 50-uL samples were compared at
307.6 nm with known standards.  Graphite furnace conditions were as follows:
nitrogen flow, 20 mL/min;  sample drying  time,  60 sec with a 15-sec ramp
(20-120°C); ashing time, 50 sec with a 10-sec  ramp (120° -500°C); and
atomization, 6 seconds at  2200°C.

     We report concentrations of PCBs and  Zn in both sediment and tissue on a
dry weight basis to alleviate discrepancies caused by varying water content.
Thus subsamples of all analytical  samples  were dried to measure water content.
A wet-to-dry conversion factor was calculated  and applied to measured
wet-weight concentrations.

     We compared PCB and Zn concentrations in  test organisms between
preexposure samples and test and control samples after exposure, using analysis
of variance (ANOVA).  For  both species of  fish and the oligochaetes, all
surviving test organisms in a tank were  composited to form one analytical

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sample.  Thus different tanks were considered  replicates.  Clams remaining
alive at the completion of the bioassay were divided  into two analytical
samples so that within- and between-tank replicates,  as well as replicate
tanks, were considered in the ANOVA.

                                   Results

     The two sediments used provided  contrasting levels of PCBs and Zn.
Concentrations in test sediment (micrograms per gram, dry weight) ranged from
12.8 to 31.7 PCB and from 147 to 241  for Zn whereas those in control sediment
were < 0.02 for PCB and from 31 to 67 for Zn (Table 1).  Although sediment was
collected three times, which resulted in some  variation in both physical and
chemical  composition, a large difference in the two contaminants of interest
between control and test sediment was always found.

     Use of the flow-through bioassay produced nearly constant conditions
throughout testing, and mortality was low (£8.3%) in test organisms,
indicating that the test sediments were not acutely toxic to the organisms
tested (Table 2). Mortality was high  in only one test where a mechanical
failure restricted flow to two tanks  holding oligochaetes during exposure.  In
these two tanks, all  oligochaetes died and were not analyzed.  Thus in the
contaminated sediment treatment only  two replicates are reported.

     All  test organisms accumulated significant (££0.05) amounts of PCBs from
test sediments when compared with either organisms exposed to control sediments
or preexposure organisms (Table 3).  Bioaccumulation factors (BF = dry wt
concentration in tissue divided by dry wt concentration in sediment) for
organisms exposed to test sediments indicated that oligochaetes were the most

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 Table 2.  Summary of test conditions and mortality  of  organisms,
Organism and      Temp.
  exposure        (°C)
(No. of organisms
in parentheses)
                                 Test Conditions
                           Dissolved
oxygen
(mg/L)
   Hardness
(mg/L,  CaCOs)
Sediment Eh
  (mV)
Mortality
Fathead minnows
 Control (10)     20.0
 Test (10)         21.2

Perch
 Control (10)     20.2
 Test (10)         20.2

Oligochaetes
 Control (14)     21.2
 Test (14)         21.0

Clams
 Control (30)     19.9
 Test (30)         19.4
  7.8
  7.5
  7.9
  7.7
  8.0
  7.9
  8.5
  8.4
   158.6
   158.6
   128.1
   128.7
   150.4
   156.0
   113.8
   113.3
 -146.3
 -175.9
 -227.5
 -219.7
 -154.5
 -214.3
 -272.5
 -271.5
  5.0
  0.0
  0.0
  0.0
  5.4
  0.0
  8.3
  8.3

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Table 3.  Mean weight,  lipid  content, and PCB and Zn concentrations in analyzed
     samples.  Standard errors  in  parentheses.  All sample sizes equal 4 except
     clams (N=8)  and oligochaetes  (N=2).
Organi sm and
exposure
Fathead minnows
Preexposure
Control
Test
Yellow perch
Preexposure
Control
Test
Oligochaetes
Preexposure
Control
Test
Asiatic clams
Preexposure
Control
Test
Weight
(9)
2.42
(0.16)
2.08
(0.08)
2.06
(0.09)
2.10
(0.04)
1.90
(0.03)
1.98
(0.09)
0.94*
(0.07)
0.64
(0.04)
0.58
(0.02)
1.05
(0.03)
1.22
(0.04)
1.12
(0.05)
Lipid
(%)
8.5
(0.29)
8.1
(0.18)
8.1
(0.08)
4.9*
(0.27)
3.9
(0.29)
3.6
(0.23)
0.6
(0.07)
0.6
(0.12)
0.5
(0.0)
1.5
(0.23)
1.8
(0.04)
1.9
(0.10)
Contaminants
PCB
1.0
(0.02)
1.4
(0.04)
45.4*
(1.96)
1.6
(0.0)
2.0
(0.18)
8.9*
(0.73)
0.4
(0.06)
0.5
(0.08)
125.5*
(1.88)
0.8
(0.17)
1.1
(0.04)
3.4*
(0.14)
(ug.g dry vrt.)
Zn
189.8
(13.36)
227 .6*
(13.34)
179.2
(3.95)
113.5
(6.92)
128.6
(8.15)
118.4
(12.79)
182.9
(24.96)
141.0
(11.47)
171.0
(36.64)
135.1
(9.64)
97.1*
(2.55)
117.8
(3.18)
*Denotes significant difference  (£_< 0.05)  from other two treatments based
 on analysis of variance and Duncan's multiple range test.
                                             10

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efficient accumulators (BF, 4)  and  clams  were the least efficient (BF, of 0.2).
Both species of fish accumulated  PCB—the BF was 1.4 in fathead minnows and 0.7
in ye11ow perch.
     None of the organisms exposed  to test  sediments accumulated Zn in any of
the bioassays, although we did  observe  several statistically significant
changes in the Zn concentration in  organisms from other treatments.  Clams
exposed to control sediment were  significantly lower (P_ < 0.001) in Zn
concentration than clams before exposure  or those held in test sediments (Table
3).  Apparently clams had high  levels of  Zn in tissues when we received them
and some depuration occurred in clean sediments.  Fatheads exposed to control
sediments had significantly .higher  (P_= 0.03) concentrations of Zn than
fatheads before exposure or those exposed to test sediments (Table 3). This
observation is unexplained.
     The weight of oligochaetes and the lipid content of yellow perch both
decreased significantly during  the  bioassay (Table 3).  The general trend of
decreasing weight and lipid content in  both species of fish and in the
oligochaetes was expected because the organisms  were not fed during the 12-day
test.
                                 Discussion
System performance
     The flow-through bioassay  that we  evaluated for use in assessing the
potential of bioaccumulation from sediments provided conditions allowing for
high survival of test organisms.  Mortality did  not exceed 8.3% in species and
half the tests resulted in no mortality.  The water flow of 100 mL/min was
                                       11

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sufficient for maintenance of  good temperature control and a high oxygen
saturation (95%).
     The 10-day exposure period, as suggested for evaluating dredged material
for ocean discharge (USEPA/CE  1977), was seemingly adequate to assess
bioaccumulation potential  for  organic contaminants with similar chemical
characteristics as PCBs.  However, it is unlikely that test organisms had
reached a steady-state concentration , thus the maximum BFs had probably not
been attained (Rubinstein et al. 1983).  We did not observe the accumulation of
Zn from test sediments in this study—perhaps because the Zn in our test
sediments was in a form that is biologically unavailable (Engel et al. 1981);
increasing the exposure time would not likely have affected the bioaccumulation
of Zn.  Rubinstein et al. (1983). showed that increasing the time that marine
invertebrates were exposed to  contaminated sediments from 10 to 100 days did
not result in accumulation of  Hg or Cd. Seelye et al . (1982) provided evidence
that 10 days is sufficient for measuring bioaccumulation potential of
biologically available metals  from sediments; they reported accumulation of Zn
and several other  metals by yellow perch in a 10-day exposure.  It thus appears
that 10 days is sufficient to  measure bioaccumulation potential of metals that
are in available form; however not all factors that  influence this availability
are understood.
     The effects of not feeding organisms during the test are not certain.  It
might be argued that withholding  food might result in a loss of PCBs as lipids
are mobilized and  lipophilic contaminants metabolized.  However, if this was
occurring to any great extent  then organisms exposed to control sediments would
have lower PCB levels than unexposed organisms.  This loss of PCBs was not
                                       12

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observed.  Nevertheless,  It  Is possible that feeding would have enhanced
accumulation and  perhaps  feeding of organisms during the test should be
attempted.  It may  not  be successful in tests with fish however due to the high
turbidity in aquaria  caused  by resuspension of fine sediments.

Species evaluation

     We compared  accumulation in two species of fish that have different
advantages for use  as bioassay test organisms.  The yellow perch 1s widely
distributed in the  Great  Lakes and the young are often found in areas of
dredging activity (Barnes 1979).  Its commercial and sport fishing value  make
it an economically  important species.  On the other hand, the fathead minnow,
although also widely  distributed in the Great Lakes watershed, is neither as
abundant nor as economically important as the perch.  It does offer certain
other advantages   (a) it  is  routinely available because it is easy to culture,
(b) it is tolerant  of a wider range of water temperature and dissolved  oxygen
(Eddy and Underbill 1974), and c) it is widely used as a bioassay organism
(Committee on Methods for Toxicity Tests with Aquatic Organism 1975).
     Our results  show that fathead minnows accumulated higher concentrations
of PCBs than did  perch.  Although this difference was not critical in testing
sediments containing  high levels of PCBs, the fathead minnow would be the
superior test organism  for testing sediments with low concentrations of
contaminants.  Higher lipid  levels in fathead minnows (Table 3), as well  as
behavioral differences  between the two species, could account for their greater
uptake of PCBs.  Fathead  minnows were more active at the water sediment
interface than were yellow perch, resulting in a greater resuspension of the
sediment.  This observation  is supported by measurements of suspended solids in
                                      13

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tanks of the two species duriny the bioassay.  In control and test sediments,
used with fathead minnows suspended solids in control and test sediments were
125.8 and 200.8 my/L,  respectively).   In the yellow perch exposure these values
were only 23.5 and 27  mg/L.

     0^. tyrtaeum was shown to  be the preferred invertebrate test species
because it accumulated PCBs  to a yreater extent than did the clams.  Althouyh
this species is not readily  available  and not a common inhabitant of Great
Lakes nearshore areas  we feel  an ol iyochaete worm would provide a benthic
invertebrate for freshwater  testiny similar to the polychaete Nereis used in
marine sediment evaluation (Rubinstein et al. 1983, Elder et al. 1979).   Both
Nereis and 0_. tyrtaeum appear  to accumulate oryanic contaminants readily and
are of adequate size to provide ample tissue for analysis.
     The Asiatic clam  is common in several areas of the country and has  been
found in western Lake  Erie (Scott-Wasilk et al. 1983).  However, our results
suygest that its suitability as a test species for measuriny bioaccumulation is
questionable due to several  factors:   (a) it may cease feediny in certain
sediments, (b) it had  the lowest BCF of all the species we tested, and (c) the
presence of the shell  causes confusion as to what to use as an analytical
sample.  Althouyh Asiatic clams accumulate metals in their shells (Clarke et
al. 1979), the ecoloyical and  toxicological siynificance of this metal
accumulation is unknown.  This leads to uncertainty as to whether shells should
be included in the contaminant analysis.  Both oryanic and inorganic analyses
are often conducted on samples taken from the same preparation, but the
inclusion of shells in the oryanic analysis could result in analytical
problems.
                                    14

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                                    References

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Barnes, M. D.  Inventory and review of  literature data sources pertaining to
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Clarke, J. H., A. N. Clarke, D.  J. Wilson, and J. 0. Frauf.  On the use of
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Eddy, S., and J.  C.  Underhill.  Northern Fishes.  University of Minnesota
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Elder, D.  L.,  S.  W.  Fowler, and G. G. Polikarpov.  Remobilization of sediment
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     Engineers Waterways Experiment Station, Vicksburg, Mississippi, 1980.  82
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Prater, B. L., R. L. Bennett, P. J. Crerar, and R. A.  Laskowski-Hoke.   An
     evaluation  and  refinement of a 96-hour sediment bioassay procedure.
     (Draft report)  EPA 68-01-6471.  U.S. Environmental Protection Agency,
     Chicago,  Illinois, 1983.  90 pp.
                                     16

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Rubinstein,  N.  I.,  C.  N.  Asaro, and C. Sommers.  The effects of contaminated
     sediments  on representative estuarine species and developing benthic
     communities.   In:   Contaminants and Sediments, Vol. 1, R.A. Baker,  ed.
     Ann Arbor  Science,  Ann Arbor, Michigan, 1980.  pp. 445-461.

Rubinstein,  N.  I.,  E.  Lores,  and N. R. Gregory.  Accumulation of PCBs, mercury
     and cadmium by Nereis virens, Mercenaria mercenaria and Palaemonetes pugio
     from contaminated  harbor sediments.  Aquatic Toxicology 3:249-260,  1983.

Scott-Wasilk, J., G. G.  Downing, and J. S. Lietzow.  Occurrence of the Asiatic
     clam Corbicula fluminea  in the Maumee River and western Lake Erie.   J.
     Great Lakes Res.  9:9-13, 1983.

Seelye, J. G.,  R. J. Hesselberg, and M. J. Mac.  Accumulation by fish of
     contaminants released  from dredged sediments.  Environ. Sci. Technol.
     16(8):459-464, 1982.
Seelye, J. G.,  and  M.  J. Mac. Bioaccumulation of toxic substances associated
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     appendices.
                                        17

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                                    TECHNICAL REPORT DATA
                            (Please read Inaructions on the reverse before completing)
i. REPORT NO.
   EPA-905/3-84-007
                                                            3. RECIPIENT'S ACCE5SION>NO.
4. TITLE AND SUBTITLE
  Flow-through Bioassay for  Measuring Bioaccumulation
  of Toxic Substances from Sediment
             5. REPORT DATE
               November 1984
             6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
  Michael  J.  Mac, Carol C. Edsall,  Robert J.
  and  Richard E. Savers. Jr.	
                                                            B. PERFORMING ORGANIZATION REPORT NO.
Hesselberg,
9. PERFORMING ORGANIZATION NAME AND ADDRESS
  U.S.  Fish  and Wildlife Service
  Great Lakes Fishery Laboratory
  Ann Arbor, Michigan  48105
             10. PROGRAM ELEMENT NO.
               Contribution No.  616
             11. CONTRACT/GRANT NO.
                                                              IAG-DW 930095-01-0
12. SPONSORING AGENCY NAME AND ADDRESS
  U.S.  Environmental Protection  Agency
  Great Lakes National Program Office
  536  South Clark Street, Room 958
  Chicago,  Illinois  60605
             13. TYPE OF REPORT AND PERIOD COVERED
               Research Report 4/83  -  5/84
             14. SPONSORING AGENCY CODE
               Great Lakes National  Program
               Office-USEPA-Region V
16. SUPPLEMENTARY NOTES
  Anthony  Kizlauskas
  Project  Officer
16. ABSTRACT
  A bioassay  was developed for  testing the solid phase  of  dredged material  that measures
  the survival  of organisms, and  the bioaccumulation of toxic substances.   This bioassay
  is intended to estimate the bioaccumulation potential  from sediments that are not
  acutely  toxic to test organisms,  but are suspected of containing persistent
  contaminants.  Two species Of fish, Pimephales promelas  and Perca flavescens, and
  two invertebrate species, Octalasion tyrtaeum and Corbicula fluminea were used in
  the evaluation of this flow-through bioassay.  Although  not intended as a toxicity
  test per se,  the test enables interpretation of any mortality that may occur  during
  bioassays.
17.
                                 KEY WORDS AND DOCUMENT ANALYSIS
                  DESCRIPTORS
                                               b.IDENTIFIERS/OPEN ENDED TERMS
                            :.  COSATI Field/Group
  Bioassessment Test
  Aquatic Organism
  Toxic Substances
  Sediments
  Dredging
19.
   DISTRIBUTION STATEMENT
  Document  is  available to the public
  through the  National  Technical  Information
  Service,  Springfield, Virginia*22161
19. SECURITY CLASS (This Report)
   UNCLASSIFIED
21. NO. OF PAGES

     26	
20. SECURITY CLASS (This page)
   UNCLASSIFIED
                            22. PRICE
EPA Form 2220-1 (9-73)
                                                   U.S. GOVERNMENT PRINTING OFFICE: 1985-555-055/497

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