v>EPA
         United States
         Environmental Protection
         Agency
               Office Of Water
               (WH-556)
EPA 823-R93-001
February 1993
Tiered Testing Issues
For Freshwater And
Marine Sediments

Proceedings Of A Workshop
Held In Washington, DC
September 16 -18,1992
                                Printed on Recycled Paper

-------

-------
            Proceedings
      Tiered Testing Issues
                for
Freshwater and  Marine Sediments
         Washington, D.C.
     September 16-18, 1992
        U.S. Environmental Protection Agency
    Office of Water, Office of Science and Technology,
       and Office of Research and Development
              Washington, D.C.

-------

-------
                           TABLE OF CONTENTS
Introduction .'....'	............>...••••••••••••••••••	

Workshop Summary	.' ' ' '	•••••• • •	•   ^
     Day One	; •  • •-.	:	• • • •
     Day Two  	v . •  • • • • • • •	 . ..	• • • •
          Marine and Estuarine Sediment Testing Break-out Session  . ,	   ป
          Freshwater Sediment Testing Break-Out Session	  21
     Day Three .	•	

                                                                     ...  37
Workplans	• • • •	
     Development of Sediment Toxicity Bioassay
     for Marine and Estuarine Organisms
                                                                         41
      Development of Sediment Toxicity Bioassays
      for Freshwater Organisms ..... ....... . ..... ....... ...... • •  ......
Outlines of Speaker Presentations .................. ...... ........ • .....
      EPA's Contaminated Sediment Management Strategy                         65
           Elizabeth Southerland  ....... ..... ....... ........ .......... 65

      Standardized Sediment Testing:
      Needs and Requirements of Key Agency Programs                            .
           Thomas Armitage ... ..... .  . . . . ......... ......... .  . ...... • 87

      EPA Regional Sediment Needs             ,
           William Peltier  .../:..... ......  ................ • •  .......

      Tiered Sediment Testing Conceptual Overview
           Elizabeth Southerland  ..... . .  ...... ............. .........

      Summary of ASTM Activities on Freshwater and Marine Sediment Test  Methods
           Chris Ingersoll  ...........••••• ..... ......... ..... ......

      EPA Approaches for Biological Methods Standardization:
      Historical Perspective and Present Guidance
           Jim Lazorchak ........... • • ..... • ..... .... ........ '. '  ' ' '

      EMMC Methods Format   .   ,   •                                       91 -a
           William Telliard  ...... ...... ............ ......... . • • ...... .4 ซo

-------
Freshwater Sediment Toxicity Assays: Necessary and Desirable Attributes
      G. Allen Burton	 227

Midge Whole Sediment Bioassays
      John P. Giesy and Jody A. Kubitz	 257

Desirable and Necessary Attributes for Freshwater Sediment Toxicity Tests:
Hvalella azteca
      Chris Ingersoll	269

Discussion of Desirable and Necessary Attributes for Marine and Estuarine
Sediment Toxicity Tests, and the Use of  Ampelisca abdita.
Rhepoxvnius abronius. Leptocheirus plumulosus. and Eohaustorius estuarius
in Marine and Estuarine Sediments
      Richard C. Swartz	\ ...... 281

Bioaccumulation of Sediment-Associated Contaminants:
Significance, Current Status, and Future
      Peter Landrum	,	 . .	295

Bioaccumulation of Sediment-Associated Contaminants:
Present Status, Laboratory Methods, and Related Research Needs
      Henry Lee II		315

Discussion of the Use of Lumbriculus variegatus
in Freshwater Sediments
      Gary Ankley	'.	-....-.'	347

Identification of Long Term Needs for Assessing Sediments
      Norm Rubinstein	".	 349

Break-Out Workgroup for Freshwater Sediment Issues: Overview of the Day
      Gary Ankley	 .	357

Development of a Standard Protocol for Testing Hyalella azteca
      Teresa Norberg-King	361

Development of a Standard Protocol for Chironomus tentans
      Robert Hoke	 387

Issues for Lumbriculus varieaatus1
      Peter Landrum	'. .	393

Development of a Standard Acute Amphipod Protocol
      Richard C.  Swartz, Michelle Redmond, Janet Lamberson
      Beth McGee, Ray Alden, Richard Scroggins, Peter Chapman	 403

-------
     Development of a Standard Chronic Amphipod Protocol    _                •

          John Scott, Ted Dewitt, Michelle Redmond, Chris Schlekat ........ v;  439


                      '-••'•                           . . . .  .	473
APPENDIX A: Workshop Agenda	• • •	


APPENDIX B: Sediment Toxicity Tests Under Development by Environment Canada  ...  483


                                                                ......  491
APPENDIX C: Freshwater Surveys  ........••••••••••••••••••••


APPENDIX D: Bibliographies for Freshwater Test Species  . .	•','''


                        . .                                    ........ 538
APPENDIX E: Workshop Participants	

-------

-------
                                 introduction

-------

-------
WORKSHOP SUMMARY
         1

-------

-------
                                Workshop Summary

      The two and one half day workshop was designed to address general issues affecting both
marine and freshwater sediment testing on day one. Break-out sessions on day two focused on
specific details and requirements for freshwater and marine tests. Day three consisted of reports
back to the full workshop by break-out session leaders on issues discussed, conclusions, research
needs, and next steps followed by an overall summary and wrap-up.
Day One

       During day one of the workshop, the following talks were presented:
Introduction and Description of EPA's Sediment Strategy
Elizabeth Southerland,  U.S. EPA Office of Science and Technology

       Dr. Southerland welcomed participants to the workshop and stated that EPA was
sponsoring this meeting to provide a forum for discussion of issues related to the standardization
of sediment bioassay test methods for cross-program use.    She  described  EPA's Draft
Contaminated Sediment Management Strategy and noted that the development of consistent
assessment methods was one of the major goals of the Strategy. An outline of the draft strategy
was sent to more than 1000 representatives of industry, state, federal governments, and various
constituent group in March of 1992.  Based on comments received from the mailing and from'
three national forums held during the spring and summer of 1992,  a final strategy will be
developed and published in the Federal Register in 1993.

EPA Program Office Sediment Evaluation Needs
Thomas Armitage,  U.S. EPA Office of Science and Technology

       Dr. Armitage described the  statutory authority and regulatory responsibility of EPA
Program Offices that could use the results of sediment toxicity tests.  Based on differences hi
regulatory responsibilities, different programs may interpret test results differently. Interpretation
of individual test results may vary among programs, but standard bioassay protocols could be
used by all programs.  . Once the method protocols have been developed, they may be used
immediately by Office  of Water programs, the Superfund Contract Lab Program, and EPA's
Environmental Services Divisions. The Office of Pollution Prevention and Toxic Substances may
begin a test rule process leading to publication of test methods in the Federal Register, and the
Office of Pesticide Programs may begin their Science Advisory Panel review process.  The
methods to be developed will also be submitted to ASTM to begin the balloting process leading
to completion of an ASTM standard method.

-------
EPA Regional Sediment Evaluation  Needs
William Peltier,  U.S. EPA Region IV, Environmental Services Division - Athens, GA

       Mr. Peltier discussed the EPA Regions'  sediment testing  needs  and reminded  the
audience that  the Regions are  important  clients for the methods  under  development.   He
discussed the  test methods that are currently available, and  noted  that there are significant
differences in test conditions among them.  He emphasized that methods must be validated arid
must be run by Regional labs, state labs and consulting labs. He also noted that test conditions
such as feeding,  age of organisms, sediment depth, and water renewal must be clearly specified
in the protocols.  If pore water is to be tested, the pore extraction method must also be specified.
He also raised the issue of reference and control organism survival acceptability, and discussed
the possibility  of using synthetic sediments for controls.   Concerning  species selection, he noted
that there  should be criteria for using regional species instead  of the species selected  for
standardization,  and  that reference toxicant  testing should be conducted.   The question of
whether  reference toxicant testing should be conducted each time a toxicity  test is done, or
whether  less frequent (weekly/monthly) testing would suffice  was discussed.  The importance
of a good QA/QC program was emphasized.  This should include  laboratory evaluation,
accreditation, and other checks on lab quality. Mr. Peltier also briefly discussed Regional needs
for bioaccumulation testing, especially data interpretation, and closed his presentation with the
reminder that  Regional and state outreach was the key to success of this program.


Tiered  Sediment Testing Conceptual Overview
Elizabeth Southerland, U.S. EPA Office of Science and  Technology

       Dr. Southerland presented an overview of tiered  sediment testing. The Office of Water
(OW) the Office of Pesticide Programs (OPP), the Office of Pollution Prevention and Toxic
Substances (OPPTS), the Office of Solid Waste (OSW) and the Office  of Emergency  and
Remedial Response  (OERR) are all committed  to the  principle of consistent tiered testing
outlined in the Agencywide Contaminated Sediment Strategy.  Agencywide consistent testing is
desirable because all EPA programs would be able to agree  on whether a sediment poses an
ecological or human health risk, and comparable data would be generated. It would also provide
the basis for uniform cross-program decision-making within EPA.  Each  program, however,
should retain the flexibility of deciding whether identified risks would trigger regulatory actions.
Tiered testing should include a hierarchy of tests with the tests in each successive tier becoming
progressively  more rigorous, complex, and costly.  Interpretative guidance must be developed
to explain how  information generated within, each tier  would trigger  regulatory action.   The
interpretative  guidance could be program  specific describing  decisions  based on  a weight of
evidence approach, a pass/fail approach, or comparison to a reference depending  on statutory
and regulatory requirements. There are currently two models of sediment tiered testing used by
EPA: 1) the Office of Water/US Army Corps of Engineers dredged material testing framework;
 and, 2)  the OPP ecological risk assessment tiered testing framework.  Tier one of the dredged
 material testing  framework consists of a review of existing chemical and biological data and/or
 an inventory of  nearby sources. In tier two, chemical data are generated and compared to water
 and sediment quality criteria. Tier three evaluation consists of acute toxicity and bioaccumulation
 testing,  and a comparison of the results to a reference area.  A tier  four evaluation consists of

-------
site-specific field studies.  The OPP testing framework consists of acute toxicity testing in tier
one followed by chronic (early life stage) toxicity testing in tier two and further chronic toxicity
testing (full life cycle) in tier three. Tier four consists of field or mesocosm testing. A tiered
testing framework has not yet been chosen for Agency wide use, but some of the components that
have been identified will be standardized as a result of this workshop. These components ar?
acute and chronic toxicity bioassays, bioaccumulation tests, chemical criteria, and any others that
may have ecological significance including benthic community structure evaluation, colonization
rate, and in situ sediment  testing within a mesocosm.


Summary of ASTM Activities
Chris Ingersoll, U.S. Fish and Wildlife Service, NFCR -Columbia, MO

       Dr  Ingersoll of the U.S. Fish and Wildlife Service presented a summary  of ASTM
activities to standardize freshwater and marine sediment test methods.  ASTM has not yet
developed any standard methods for sediment testing but has developed guides. The most recent
of thesTguides include:  "Standard  Guide for Collection, Storage Characterization  aid
Manipulation  of Sediment for Toxicologies!  Testing." (ASTM  1991  Method E1391-90);
 "Standard Guide for Conducting Static Acute Toxicity Tests Starting with Embryos of Four
 Species of Saltwater Bivalve Molluscs." (ASTM 1991 Method E724-89); "Standard Guide for
 Conducting Acute Toxicity Tests with Fishes, Macroinvertebrates, and Amphibians -(AbTM
 1991  E  729-88)-  "Standard Guide form Conducting Sediment Toxicity Tests  with Marine,
 Estuarine and Freshwater Invertebrates." (ASTM 1991 E-47); "Standard Guide ^Conducting
 10-day Static  Sediment Toxicity Tests with Marine and  Estuarine Amphipods.  (ASTM 19yi
 Method  E1367-90);  and "Standard  Guide for Conducting  Sediment Toxicity  Tests with
 Freshwater Invertebrates." (ASTM 1991 Method E1383-90).  Dr  Ingersoll  .discussed the
 differences between ASTM guides and methods, and briefly described the guides listed above.


 Approaches for Test  Standardization;
 Historical Perspective and Present Guidance                    • . •  ;
 Jim Lazorchak, U.S. EPA Environmental Monitoring and Systems Laboratory - Cincinnati, OH
 William Telliard,  U.S. EPA Office of Science and Technology

       Dr Lazorchak described a 1987 ORD document, "Guidance on Methods Standardization"
 which was never .finalized but may serve as a framework for the methods to be standardized as
 a result of this workshop.  There is no specific standardization process for biological methods
 in the document,  but the process developed for chemical testing may be applied to biological
 testing methods development.  As outlined in the document, method requirements and data
 quality objectives must first be established. Method selection and development is men followed
 by a single-laboratory evaluation involving a precision check and tests for sensitivity of method
 variables. This is followed by confirmatory testing by a minimum of three labs.  An mtenm
          .
  method description may then be prepared, although this has not yet been done for a^iological
  test.. A formal collaborative or round robin testing procedure may then be conducted with a
      .
  minimum of six labs.

-------
       Dr. TelJiard briefly described the activities of the EPA's Environmental Monitoring
Management Council (EMMC) whose charter is to: 1) coordinate the Agency's environmental
methods research and development activities; 2) foster consistency and simplicity in measurement
methodology across regulatory  programs;  3) facilitate cooperative efforts with other federal
agencies, academia, industry, and other interested external parties on methods development; 4)
promote'and facilitate the adoption of new monitoring technology and instrumentation; and 5)
evaluate the feasibility of a national laboratory  accreditation program.   He  described the
activities of the Methods Consolidation Workgroup.  Their focus to date has been on methods
for water, solid waste,  and  air, although QA/QC and biological methods will be considered by
the group as well. A method validation process has not yet been developed, but a format for
EMMC approved methods has been finalized.                           /


Desirable and Necessary Attributes for  Freshwater Sediment Toxicity  Tests
G.  Allen Burton, Wright State University - Dayton, OH
John Giesy, Michigan  State University - East Lansing, MI
Chris Ingersott,  U.S. Fish and Wildlife Service, NFCR - Columbia, MO

         Dr. Burton noted that desirable attributes, for freshwater species and tests include:
species sensitivity; reproducibility of the  test; discriminatory  function of the test; and  "do-
ability". Dr. Giesy remarked that compromises are often necessary when deciding which species
to standardize.  Generally, a battery of tests with several species is required to explain most of
the variation in a test.  Several disadvantages to the use of Chironomus tentans in sediment tests
were described:  these insects are relatively insensitive for use in acute tbxicity tests (they are
especially  tolerant to  metals);  they  do not feed  directly  on sediments but eat  resuspended
particles; genetic drift has been observed in lab-to-lab variation; there  is unexplained sporadic
loss of vigor in culture; and there is the potential for loss (pupation or emergence) of adults.
Several other issues were identified:  when to, start the test; test duration; endpoints; volume of
test sediments and water;  food/culture medium.  Similar questions about test conditions for
Hyalella azteca were posed. Many of the issues highlighted by these speakers, will be described
in  the freshwater break-out session notes.


Desirable and Necessary Attributes for Marine and Estuarine
Sediment Toxicity  Tests
Rick Swarfz, U.S. EPA Environmental Research Laboratory - Pacific Division  ,

        Several  of the same attributes and issues identified for freshwater bioassays  were
 described for marine/estuarine tests.  A major difference between marine and freshwater testing
 is  that many marine test species are  field collected not cultured.  Adult organisms are used! for
 tests rather than the young cultured organisms used in freshwater testing. This difference aftects
 feeding regime  as well as the necessity for performing routine reference toxicant testing.

-------
Desirable and Necessary Attributes for Freshwater,  Marine  and Estuarine
Bioaccumulation Tests                                               mn
Peter Landrum NOAA Great Lakes Environmental Research Laboratory - Ann Arbor, MI
Henry Lee II, U.S. EPA Environmental Research Laboratory - Pacific Division
      The two required attributes of bioaccumulation test species are that they ingest sediment

                --SMS?
 i^s^^rt^^
          '      residue levels, and the need for interpretative guidance on test results.
       Attributes of Lumbnmte variegatus that make it a. good ^f f01*^


 usiln long-term tests; standard culture and test methods have been developed; and some field
 validation has been done.


 Identification of Long Term Research Needs            _ ••
 GaryAnkley, U.S. EPA Environmental Research Laboratory -Duluth, MN
 No^n Rubimtein, U.S. EPA Environmental Research Laboratory - Narragansett, RI
       Several long term research needs for sediment toxicity testing were ide ntifi ed. These


  L Development Contaminated Sediment Research Strategy -is include in the end of the manne
  break-out session summary.

-------

-------
Day Two

Marine and Estuarine Sediment Testing Break-out Session
                           -   '              .          -                      >          !
       During this break-out session of the workshop, session leaders facilitated discussion of
issues to be resolved for marine and estuarine sediment testing.


Objectives of the Session                                 •„-   '"     m'r''
Norm Rubinstein, U.S. EPA Environmental Research Laboratory - Narragansett, Rl

       Dr Rubinstein identified three overall objectives for the workshop session dealing with
sediment tests for marine and estuarine organisms:  1) the workshop participants should agree
upon the definition of a standard method; 2) the workshop participants should reach^ agreement
onthe test species and protocols that can be standardized within the next year; and 3) workshop
participants should reach agreement on the process for  standardizing protocols on sediment
spiking, handling, and storage.


Discussion of Marine and Estuarine Test Methods
Rick'Swartz, U.S. EPA Environmental Research Laboratory - Newport, OR

 Standardization Process

        Dr Swartz noted that, since exact guidance for standardizing test methods has not been
 developed, the ASTM process for standardizing methods should be used to provide peer review
 and more general participation  in the process.


 Identification of Uncertainties to be Addressed in  Standardization Process     ;

        It  was  suggested that  methods for sediment handling and manipulation should be
 addressed separately from the toxicological methods .themselves.  It was noted that Environment
 Canada is evaluating sediment handling and manipulation as a separate issue, and will soon have
 guidance  available. There was general agreement that  guidance is needed to determine how
 sediment  should be sampled and handled in the field, transported or shipped to the laboratory
 and manipulated or otherwise treated in the laboratory. It was suggested tot handling;imd.
 spiking could be addressed in guidance documents to accompany test method documents.; There
 was also discussion about developing an additional standard method for experimental design that
 could provide specific information on such issues as sampling replicates and selection, of held
 reference sites.

-------
       Dr. Swartz identified three categories of uncertainty that must be addressed in developing
guidance:  1) limitations of the method must be described (this is distinct from information
gaps); 2) issues that can be resolved on the basis of consensus should be identified (e.g. selection
of the temperature for conducting tests); and 3) critical research needs that must be met should
be identified (e.g. grain size tolerance).  Workshop participants agreed to identify these issues
for the available test organisms.
Discussion of Rhepoxvnius dbronius
Rick Swartz, U.S. EPA Environmental Research Laboratory - Newport,  OR

       Dr. Swartz indicated that Rhepoxvnius has been well studied, there are 40-50 papers in
the literature, and there is already one interlaboratory comparison that has been completed. Dr.
Swartz  noted that  there are  probably not many critical research  needs  to be rilled for
Rhepoxvnius tests.  The following consensus issues were identified for Rhepoxynius:

1.     A written protocol for  the standard acute toxicity test is needed.  Testing procedures
       which must be followed should be identified.

2.     Agreement must be reached on the data needed for reference toxicants.  Dr. Swartz
       already has data on cadmium.

3.     Reference sediment quality  assurance/quality control  requirements must be identified.
       Specifically, the question of the need to set a minimum desirable reference sediment
       survival limit was raised. If one has intermediate levels of survival in reference sediment,
       the difference between reference sediment and test site sediment, if detected, are difficult
       to interpret,  if not meaningless.

4.     Data are available on the seasonal sensitivity of Rhepoxynius.  It should be possible to
       understand seasonal variability of this test species.

5.     Consensus must be reached  on how to ship and handle Rhepoxynius.

       The need for evaluating relative sensitivity of Rhepoxynius was  discussed. Reference
toxicants were discussed, and it was agreed that the method guidance developed for Rhepoxynius
should include information describing how to interpret sensitivity to reference toxicant tests.
The utility of reference toxicants spiked in sediment was addressed. Workshop participants
stated that this is a generic issue to be addressed in all of the method guidance documents to be
developed.
                                          10

-------
Discussion of Ampelisca dbdita
Michelle Redmond, U.S. EPA Environmental Research Laboratory,- Newport* OR

       Ms Redmond described the test species and the, acute toxicity test.  Ampelisca abdjta
occurs in the high salinity range of estuaries.  It is a particle feeder. Research to support the
Ampelisca test is needed in a number of areas:

. 1.     Research is needed on grain size tolerance for this species.

2.     No data are available to test the sensitivity of ovigerous females.  Males are not used in
       the test.

 3.     A critical research question is how to interpret control survival. Frequently problems are
       encountered when laboratories run the test for the first time.

       Other issues were discussed. It was noted that problems have been encountered when
 Ampelisca is shipped to laboratories to run the test. The shipping process must be standardized.
 Test sensitivity has also been observed related to season. It was noted that EPA's Environmental
 Monitoring and Assessment Program has adopted a control survival limit of 90 percent for this
 species   For Hyalella a lower  rate of 80 percent has been established.  Salinity ranges for
 AWlisca were also discussed.  Some workshop participants recommended using full strength
 seawater for the test.  The species was not recommended for tests at salinities below five parts
 per thousand Workshop participants agreed that a standard method should be written for a high
 salinity range, and additional research can be conducted to broaden the range.  There was some
 discussion of ammonia tolerance,  and  it was agreed that  recent research has established the
 ammonia tolerance limits of Ampelisca.


 ntsni.ssinn nf T^ptocheirus plumulosus
 Beth McGee, Maryland Department of the Environment - Baltimore, MD

        Ms McGee described tests using L^tocheirus.  She noted that it has a wide tolerance
 range for both salinity (2-30 parts per thousand) and grain size (sand to silt). It was agreed that
 test temperature is a consensus issue, not a research issue for this species.  The test is run at
 temperatures between 20-25 degrees C. The salinity at which, tests are run should depend upon
 the objectives of the test.  When testing the toxicity of in-place pollutants, the test site salinity
 may be used for the test.  A photoperiod of 16:8 hours light/dark is used for the test.  Juvenile
 and young adults (sized 3-5 mm)  are tested and 20 animals are used per test chamber.  Both
 field collected  and  cultured organisms are used for the test.  Cadmium chlonde is used as a
 reference toxicant for the test (96-hr LC-50).
                                            11

-------
       A number of research issues were identified for Leptocheirus:

1.     What are the effects of salinity on test results?

2.     Must the animals be acclimated if culture or collection site salinity is different from test
       salinity?  Tests suggest tolerance limits for sudden and extreme (i.e. 5 to 32 parts per
       thousand)  salinity changes.

3.     Are different test results  obtained due to differences in •culturing methods, acclimation,
       and salinity?                          '        .

4.     More data are needed to  determine the sensitivity of this species to different chemicals,
       particularly  ammonia and PAHs.

5.     More interspecies comparisons are needed.  Data from tests on this species could be
       compared to marine species, and to freshwater species capable of tolerating low salinity
       (i.e.  Hyalella azteca).

6.     More data are needed to identify sensitivity differences between laboratory and field
       collected animals.

7.  '   Field validation is needed.

       It was noted that there are some advantages to laboratory culture of this test species. In
culture it is available at  all times of the year, and cultures  have been reared under known
conditions.  The disadvantages associated with laboratory culture include: the deleterious effects
of inbreeding and the influence of culture conditions on test sensitivity.  Advantages associated
with field collection include:  testing a natural population; the availability of large numbers of
animals at low cost; the seasonal availability of size classes; accounting for the seasonal effects
on sensitivity; and  accounting for geographic differences in sensitivity.

       A number of positive attributes of Leptocheirus as a test animal were discussed:  1) the
animal is tolerant of a wide range of environmental conditions;  2) the animal has sensitivity
which is comparable to other amphipods; 3) the animal is hardy and tolerant of handling, and
it can  be well maintained in the laboratory; and, 4) a partial life cycle test using Leptocheirus
is now being developed. It was  noted that investigators should be careful not to release animals
in non-native regions. Workshop participants agreed that indigenous species should be used in,
testing when this is possible.

Note:  An interlaboratory comparison has been conducted. Four laboratories participated in the
acute  I- plumulosus portion of a broader comparison  of test methods for Chesapeake Bay
methods.
                                            12

-------
                       ous                                T        __
                               vnentvl Research Moratov - Ne^on, OK
                       A o^to tn-riHtv test using Eohaustorius were described. The species

in the spring. The young ™"*** eis nojrowe m    g   g            ^
                                                   AW	       used with this
species is cadmium chloride.
       The following research issues were identified Eohaustorius:
 1.     Research is needed to de^^^
 2.     The response to cadmium has varied.  Additional data are needed.
 3.     /Additional'research is needed on factors affecting the sensitivity to other reference
       toxicants.
 4.     The toxicity test has not been subjected to interlaboratory  comparison.
 5      The toxicity test has not been subjected to rigorous field validation.


              to'the use of the species is that it cannot be cultured.         ,
                                    ***ซ

                               ซ
                                            13

-------
  to toxicity testing in the fall, it seems to be sensitive to organic toxicants, and is tolerant of a
  wide range of salinities. Sediment particle tolerance may be a problem.  Lower survival in fine
  sediment has been noted. Subadults are used for testing with 20 animals placed in each exposures
  chamber.

        This animal appears to be a good candidate test species for acute testing but not for
  chronic testing, since it is slow in growing and  reproducing.  The amphipod is tolerant  of
  ammonia but more research is needed  to define  its tolerance levels.  A 16:8  light to dark
  photoperiod is used in most testing with this species. This has been used to simulate summer
  conditions.  The Chesapeake Bay Program has been running tests with this species at a salinity
  of 15 parts per thousand, reference toxicant testing is conducted at 20 parts per thousand.  The
  appropriate photoperiod for testing was discussed.  It was noted that when the lights are turned
  off animals come out of their tubes and  may be exposed to more fresh  sediment.  The
  availability of the species in the field was discussed. They are a dominant amphipod  in North
  Carolina and South Carolina, and are also readily available in the Chesapeake Bay.


 Discussion of Canadian Experience -with Test Methods
 Richard Scroggins, Environment Canada - Quebec, Canada

        The Canadian experience with toxicity testing for that country's ocean dumping program
 was discussed. Seven amphipods, all found in Canada, were evaluated in a series of round robin
 tests.  The testing resulted in similar responses  at all laboratories for a number of species.   It
 was mentioned that Environment Canada will recommend that organisms for testing for Atlantic
 and Pacific sites will be coast specific.  The preferred animals for testing on the Pacific coast
 appear to be Rhepoxynius or Eohaustorius. For tests on the Atlantic Coast, Leptocheirus pjnouis
 was selected.
 General Discussion of Acute Amphipod Tests

       A number of issues were discussed by workshop participants:
I.
Experimental Design
       Guidance on experimental design should be provided to support the test method guidance.
A separate document could  be developed  that  would address  experimental  design and
interpretation of data.  Workshop  participants noted that  it is difficult to separate effects
observed in the lab from those that may or may not be observed in the field.  Richard Scroggins
recommended the use of five field replicates for conducting sediment tests.  It was noted that
appendices could be prepared  for the test methods by program  offices indicating how their
regulatory needs could be met by the tests.
                                          14

-------
II.     Commercial Availability of the Species
                                                                       •*>,
       Workshop participants discussed the importance of commercial availability of test species.
Ampelisca is available from a commercial supplier on both the east and west coasts. There are
also suppliers for Eohaustorius and Rhepoxvnius. Some workshop participants indicated that a
directory of suppliers for test animals would be very useful. .

m.    Sediment Handling and Storage

       It  was  agreed  that  the sediment  handling  and  storage issue is very  important:
Geochemists should  assist in the development of guidance in this area.  Sediment handling
jnvolves a number of uncertainties.  The Canadian government is trying to build on  the 1990
ASTM guidelines and has formed  a subcommittee  of geocherhists and field  collection
lexicologists to refine the document. The issue of storage is also important. The EPA Regional
people at  the session indicated that the commercial aspects of sample handling and storage are
important as well.

IV.   Other Issues Related  to Acute Tests

       Some participants noted that a separate guidance document  addressing experimental
design and providing guidance on sampling would be useful.  It was stated that the Regions and
Program Offices should provide input to address this issue.   A number of comments on acute
test method development were received from the Regions and Program Offices.

        1..    Superfund Enforcement indicated that, generally, they do hot have problems using
              ASTM methods, and supported pursuing ASTM approval of methods developed.

        2.     OPPTS  indicated that their real interest,is in spiked sediment tests. '

        3.     Region  1 indicated that methods developed should be subjected to interiaboratory
              validation.

        4.     Region  9 (represented by Brian Melzian  currently at the Narragansett ERL)
              indicated  that  the development  of methods should be  accompanied, by a
              technology transfer effort.    '.,_••„   .'     .  •

        5      Region  4 indicated that they  are looking for standard methods that can be
    ,     '    customized to meet a particular program need. The Region will probably use a
               set of reference toxicants to evaluate methods.
                                            15

-------
Discussion of Chronic Test Methods
John Scott, Sciences Applications International Corporation - Narrdgansett, RI

       A number of advantages  to the use of chronic tests were identified. The tests are more
sensitive than acute tests, and they may be more relevant to ecological processes.  They may
provide a better estimation of population  level effects.   A number of technical issues were
identified that must be addressed to complete development of the chronic tests. These included:

1.     Standardization of the age of species to be used.

2.     Understanding species sensitivity in chronic tests.

3.     Understanding nutritional requirements of test species.

4.     Developing feeding protocols.                     ,

5.     Understanding effects of sediment aging.

6.     Selecting the proper endpoints and biological responses for testing.

       Ted Dewitt described four species indigenous to Chesapeake Bay that were evaluated for
chronic tests.  He indicated that Leptocheirus appears to be the most promising one for chronic
tests at this time.  The species is sensitive, it reproduces easily, and it can be handled easily.
The animals can be tested in "dishpans" using static renewal conditions.  A water bath is used
to keep the test temperature constant, and an algal (a flagellated chrysophyte) and dry food
mixture is used  to maintain the animals.              ,

       It was  noted that in developing chronic tests, a procedure for minimizing the release of
nonindigenous species should also be developed.  One of the differences between a chronic test
and a 10 day acute is the requirement for feeding of animals in the longer test.  A number of
endpoints for the chronic test were discussed. These include:

1.     Mortality of adults.

2.     Size of F0 generation.

3.     Fertility  (number of offspring/female survivor).

4.     Timing of brood emergence.
                                                                \
5.     Reburial  timing.

6.     Growth.
                                           16

-------
An advantage of using ^BafitetaS in chronic tests is that the animal can survive a range of
sediment grain sizes and salinities.

       m.    ^MpVat  of the  Maryland  Department  of the  Environment  discussed  the



laboratory.










 is developed:                      ,

  1.    Appropriate test temperature.              .                       .;'.'"-
  2.    Appropriate age (U,< 24 hours xs  < 1 week old) of animal to be used for fte^.

  3.    Effect of source of animal on the test.

  4.    Effects of using cultured versus field collected animals for the tests.

  5.     Effects of the feeding regime used for the test.

  6.     Development of consistent QA/QC guideUnes (reference ;toxi^^
         survival).
  >7.     interaction between1'nutrition and toxicological sensitivity.

   8.     Effects of other variables on the test (e.g. salinity and grain size),

   9;     Simplification of the memcdolo^

    10.   Comparison of ^relative sensitivity of endpoints between species.
                                              17

-------
        Michelle Redmond discussed the use of Ampelisca in chronic testing.  This species has
 a growth curve similar to Leptocheirus.  A number of experiments have been conducted  to
 improve the reproduction of this animal in the laboratory.  The effects of differences  in
 population density, aeration, and other variables has been investigated. Success in maintaining
 the ^cultures has been variable.  Experiments indicate that aeration is necessary to conduct the
 sediment test, and that survival of juvenile offspring obtained from cultured females is greater.
 It will be necessary to look at shipping and handling to determine if this is related to decreased
 survival.  The following observations were made based on test results:
 1.

 2.

 3.

 4.


 5.

 6.
Short term and chronic sublethal tests can be conducted using Ampelisca.

Known age animals can be released from the females.

Newly released or 10 day old animals can be used for the test.

Problems of low reproduction and poor survival were encountered, perhaps related to
shipping  and handling.

A flow through system and changes in photoperiod may be necessary to conduct the test.

Nutrition may be a factor affecting test results.
Discussion on Bioaccumulation  Testing
Henry Lee,  U.S. EPA Environmental Research Laboratory - Newport, RI

       The development of a standard  28 day bioaccumulation method for two species, Neries
and Macoma, was discussed.  A draft guidance manual on bedded sediment bioaccumulation
tests has been produced by the EPA Newport Laboratory.  Workshop participants agreed that,
given the current state of knowledge, these test protocols can now be written.  Some additional
research must be completed.  Longer  term tests and kinetic models will also provide tools to
evaluate bioaccumulation.  It was recommended that a series of round  robin tests be complete
using Neries and Macoma to provide some indication of the precision to be obtained with the
tests.  The round robin experiments will not be cheap because of the requirement for tissue
analysis. It would probably be necessary to use spiked sediment with a high Kow compound.

       Region 9 has expressed interest in using'their own species for the bioaccumulation test.
Workshop participants noted that it will be necessary to develop guidance on interpretation of
bioaccumulation results. This will depend upon whether human health or ecological health is
of concern.  Human health may be the most significant endpoint to be addressed.
                                         18

-------
      It was agreed that the protocol will be developed as follows:


I/    Protocol will be for a 28 day solid phase test.


2.    Test will use a flow through seawater system.


3     Test protocol will describe test organism acclimation, maintenance, introduction, data
      recording, removal, gut depuration, and sample preservation.


      With this protocol, 80 percent of the steady state level will be reached  There was some









 Discussion  of Office of Research  and  Development Contaminated  Sediment


                     EPA EnrirovneMol Research Laboraory - Narraganse,,, RI
       The EPA Office of Research and Development (ORD) *ntamina^ฑd"S^
 ซmfซv vL discussed  ORD has restructured the research planning process   Contaminated






 STon STe "^Hbrium Phoning approach for the development of sediment quahty,

 criteria   Research ^ill also be J^conducted on evaluation of ecological nsk.
                                          19

-------
20

-------
Day Two

Freshwater Sediment Testing Break-Out Session

       During this break-out session of the workshop, session leaders facilitated discussion of
issues to be resolved for freshwater sediment testing.



Objectives of the Sessions               t'   ซ.  ^ i  *  ป™
Gary Ankley, U.S. EPA Environmental Research Lab - Duluth,  MN

       As a preliminary step for the second day of the workshop, a questionnaire was sent out
to worklop'SLSd o*er selected researchers on July 13, 1992. The pnm*y ^formation
reauested was about culturing and testing for three freshwater species OL azteca,  C ten^s,
STrofeSS T'Hie survey was done in order to assemble as much information  a^ possibly







 exampLSe culturing issues ranked for IL azteca were: known age cu tiire systems; feeding
 rSmt;' w^r for coring; flow-through versus static V^-.^'S^K
 control  e.g., reproduction levels, reference toxicants); and genetic dnft/^^ej^s -™
 testing issues for Hvalella were:   test length/endpoint; organism age to stort the tes t  water
TeS ^frequency ofc method); interpreting the effect of sediment vanable on test r^
  rg!orgSc carbon, particle size); feeding levels/appropriateness in ^^^^
 Ssurkncl/quality control  (criteria for acceptable tests).  These were generally the same issues
 for C.. tentans and I- vanggatui.            ,-.•                      •

        This session was designed to: (1) identify freshwater sediment toxicity tests as candidates
  for stan^dStion Sn th! next year; and (2) to explore specific: technical issues associated
  w^^rjpurposes of reaching a general consensus or for ซf^*
  future research.  Each issue identified during the session was discussed and, where
  consensus was reached for development of standard protocols.
  Development of a Standard Testing Protocol for mS^em
  Teresa Norberg-King, U.S. EPA Environmental Research Lab, - Duluth, MN

         Twenty one responses to the survey were received,  and  eighteen laboratories  (see
  AppendbTS reported formation on HyaOeUa aztecaf The summary of the survey responses
  are as follows: .

  I      c.^^rYfarffwzfeZfaoaeai.-- The most common procedural response is underlined and
         when no item is underlined it indicates no single most common response.
                                            21

-------
A.     Culture Methods
      Flow:
      Temperature:
      Light:
      Chamber:
      Age animals:
      Freq. restart:
      Water  Quality:
      Source of Strains:

      Aeration:
      Feeding:
      Substrate:
      Reference
      Toxicants:
Static vs. renewal
19 to 25ฐC (23!Cy
16:8 photoperiod; 50 to 100 ft. candles
!Lto40L
Known age vs. mixed age
Monthly, every 2 months
Natural vs. reconstituted
ERL-Duluth, ERL-Corvalis. Burlington, Michigan  State (most
cultured in moderately hard water or hard water)
Moderate
Leaves.  Tetraminฎ. rabbit chow,  diatoms, yeast, wheat grass,
Chlorella, alfalfa, Nutrafinฎ,  YCT. paper towels, Selenastrum,
Ankistrodesmus, brine shrimp, aquatic plants, sediment.  Feed 2
to 3 times/week typical.
Leaves, nylon mesh, cotton gauze,  3-M web plastic, paper towels

Cd. Cu, KC1, Zn, NaCl, Cr (water-only exposures)
      B.  Testing Procedures
      Flow:
      Aeration:
      Temperature:
      Light:
      Chamber:
      Sed. ratio:
      Age animals:

      No. animals:
      No. reps:
      Duration:
      Feeding:
      Endpoints:

      Acceptability:
Static vs. renewal.                                  .
None or moderate
20 to 25ฐC (2Q1Q
16:8 photoperiod; 25 to 50 ft. candles"
30 mL to 1 L (250 to 300 mL)
1:1 to 1:4 ratio sediment:water
Known age (0 to 7 d, 7 to  14 d) vs. mixed age (size about 7
to 14 d) (sieved)                                          '
5 to 20/beaker (10/beakert
2 to 10/treatment (3 to 5/treatmenfl
2-1028^(10^)
None, Rabbit Chow, YCT, maple leaves, Tetraminฎ
Survival, length,  weight,   sexual  maturation  (males),  young
production, bioaccumulation
Survival (80%). length, weight
                                         22

-------
Development of a Standard Testing Protocol
for Chironomus tentans
Robert Hoke> AScl - Duluth, MN

       Twenty one responses to the survey were received, and twelve laboratories (see Appendix
D) reported information on Chironomus tentans.  The summary of the survey responses are as
follows:
       Summary for Chironomus tentans. The most common procedure is underlined and when
       no item is underlined it indicates no suigle most common response.

       A.  Culture Methods

       Flow:              Static vs. renewal
       Temperature:        19 to 25ฐC (2310
       Light:              16:8 photoperiod; 50 to 120 ft. candles
       Chamber:           1 L to 40 L
       .Age animals:        Known age vs. mixed age
       Freq. restart:        2x week to every 6 months
       Age restart org:     egg cases to <24 h old larvae
      'Water Quality:      Natural vs. reconstituted
       Aeration:           Moderate
       Feeding:            Tetraminฎ. Nutrafinฎ, YCT and algae, alfalfa and Tetramin
                          Feed daily to 3x/week                                  .
       Substrate:          paper towels (bleached or unbleached); sand
       Reference                           •
       Toxicants:          Cu, NaCl, Cd, KC1 (water-only exposures)
       B.  Testing Procedures

       Flow:              Static vs. renewal
       Aeration:           None or moderate
       Temperature:       20 to 25ฐC (23ฐC)           ;
       Light:              16:8 photoperiod; 25 to 120 ft. candles
       Chamber:          50 mL to 2 L
       Sed. ratio:          1:1 to 111 ratio sediment:water
       Age animals:       Known age (0  to 16 d; 10 to 14 d)
       No. animals:  -     15 to 80/beaker (10 to 15/beaker)
       No. reps:          2-15 (3 to 4)     .   • .
       Duration:          2 to 14-d (10-d)
       Feeding:,          trout chow, Tetrafinฎ, YCT
       Endpoints:          Survival, weight
       Acceptability:       Survival (70%). weight (dry weight)
                                         23

-------
Development of a Standard Testing Protocol for
Lumbriculus variegatus
Peter Landrum, NOAA Great Lakes Environmental Research Laboratory - Ann Arbor, MI

      Twenty one responses to the survey were received, and five laboratories (see Appendix
D) reported information on Lumbriculus variegatus.  The summary of the survey responses are
as follows:
I.      Summary for Lumbriculus varieeatus.  The most common procedure is underlined and
       when no item is underlined it indicates no single most common response.
       A. Culture Methods
       Flow:
       Temperature:
       Light:
       Chamber:
       Age animals:
       Freq. restart:
       Water Quality:
       Aeration:
       Feeding:
       Substrate:
       Reference
       Toxicant:
Static vs. renewal
22to24ฐC
16:8 photoperiod; intensity unspecified
1 L to 40 L
mixed
Monthly, every 2 months
Natural vs. reconstituted
Moderate
Frozen silver cup trout chow, salmon starter, sediment, Tetraminฎ,
yeast, wheat  grass, Chlorella, alfalfa, Nutrafinฎ,  YCT, paper
towels food.  Feed  2 to 3  times/week typical.
paper towels, sediment

no reference toxicants specified
       B. Testing Procedure
       Flow:
       Aeration:
       Temperature:
       Light:
       Sed. Ratio:

       Age animals:
       No. animals:

       No. reps:
       Duration:
       Endpoints:
       Feeding:
       Acceptability:
Static vs. renewal
None or moderate
10to23ฐC
16:6 photoperiod
1:1 to 1:4 ratio sediment: water (sediment volumes should be,
adequate to allow feeding and .burrowing)
Adults, 3.8 cm.
Adequate number to provide tissue mass for analysis of
residue of concern
4 to 5/treatment
10 to 28 d
Bioaccumulation
None
Adequate tissue mass for residue analysis
                                         24

-------
Workgroup Recommendations
                                                                 I -•   i
       Following the presentations of the culturing and testing survey results, there was an open
dialogue about the key issues.  Generally for most of these issues,  workgroup participants
offered suggestions about currently used versus preferred approaches.  The workgroup arrived
at a consents on several culturing and testing specifics. Where it was not possible to make a
decision because of lack of information,, the group identified research items that need further
consideration before a  specific decisions could  be made.  The workgroup's consensus and
research issues for the various species tests are summarized below.

       In developing guidance for culturing freshwater species  to be included in the EPA
methods manual for sediment tests, it was generally agreed that *ere^f/^tU^fofmt
that may be used to culture the three species. It was generally concluded that success of the
tests  would rely heavily on the health of the culture from which the animals were taken for
testine  That is having healthy animals of known quality and age for testing was deemed to be
Sfkey ^deration Relative to  culture conditions.  Therefore, a perforrnance-based cntena
approach was selected  as the preferred method  through which individual labฐrat^s^
evaluate their culture protocol rather than by a control-based cntena approach.  This method
was chosen to allow each laboratory to optimize their own, perhaps umque,  culture techniques
and meet certain quality control  monitoring steps in cultures,  while providing organisms that
 would produce  reliable, comparable test results.
 Tfvalelia azteca:
 Performance-based culturing criteria, for Hyalella azteca would .include:

 1      Laboratories must perform .monthly water-lily reference toxicant tests to assess the
        health of their culture organisms or the organisms they purchase from other laboratories.
        The reference toxicant test should be performed as a 96 h water-only test.  laboratories
        should also evaluate the slope of their LC50 curve for  each reference toxicant test
        Results of these monthly tests then, would be entered into "control" charts; test results
        greater than two standard deviations from the mean LC50 might indicate a cause for
        concern that the cultures are unhealthy.

 2     Laboratories should track  the parental survival because it is important to know the
   ^     reproduction trends of reproduction  for  the cultures; it was, suggested  that this; be
        developed in a manner similar to that used as the control chart  for reference toxicant
        data.                                            '

  3 •    Laboratories should routinely measure and record the following culture water chemical
        parameters: pH; D.O.; hardness; alkalinity; and ammonia.  Again it was suggested_ tha
        {Ssbe developed in a manner to that used  as  the control chart for reference toxicant
        data.
                                            25

-------
4.     Laboratories should characterize and monitor the quality of the food they use in terms
       of nutrient content and contamination.

5.     Laboratories  should  keep records of their culture restart interval and keep accurate
       records on the age of the brood animals and, as far as possible, track the source of test
       animal to the age of the brood animals.  Physiological parameters such as lipid content
       also might be considered as a culture parameter to determine the health of the organisms.

6.     Laboratories should develop standard operating procedures to ensure that their cultures
       are renewed and monitored on a regular and standardized manner.


       Given that the performance-based culture criteria will be part of the requirements for the
test performance, the major consensus  items for the Hyalella  azteca  test protocol were  as
follows:

1.     The use of the performance-based culture  criteria for cultures will be used to judge the
       health of the animals in the  toxicity test, along with the acceptability criteria. The test  (
       acceptability criteria were agreed upon as  80% control survival in a 10 d test as well as
       acceptable water chemistry parameters over the course of the test.

2.   "  The renewal  of the overlying water in the  sediment  test at a rate of  1.25  to  4
       volumes/day was agreed upon.  The specific procedures were not specified for the water
       renewals; however, the intervals  should be evenly spaced over 24 h.

3.     The age of the animals for testing was 0 to 14 d. However, the workgroup recognized
       that older animals (7 to 14 d) are easier to recover in the whole sediment compansd to
       0-7 d old animals.  This necessitates the need for known age culture systems.

4.     The test length discussed ranged  from 7-28 d,  yet since the majority of the workgroup
       participants were using 10-14 d for the survival endpoint, consensus was to use the 10
       d test period. This test duration may allow the growth endpoint to be measured after 10
       d, but more research "would be needed before this could be added as a criterion for test
       acceptability.

5.     The number of animals needed per concentration and the number of replicates needed was
        discussed. The choice of each (replicates, number per concentration) will depend in part
     •  on the probability level selected, and the type of statistical  analysis.  When variability
        remains constant, the sensitivity of the test increases as the replicates increase. Further
        evaluation of the appropriate alpha  (a) (significance level)  and beta  (6) (power of the
        test)  for the Type I and Type n errors is needed along with consideration of the delta (5)
        (minimum detectable difference).  It was the group consensus'that this analysis could be
        done with existing data.
                                            26

-------
6      Although various size test chambers are used, it appeared that numerous researchers were
       looking for smaller test volumes and test chambers and use of 250 to 300 ml beakers
       with 50 to lop ml of sediment for 10 to 20 animals each was agreed upon.  Larger
       chambers have been used in the past but because space can be a limiting  factor or
       recovery of animals from larger volumes of sediment may be lower, it was the groups
       consensus that smaller chambers would be acceptable. The minimum amount of sediment
       needed to use in tests may need further consideration.

7      Feeding during the 10-d survival exposure was deemed necessary. Otherwise, acceptable
       survival cannot be achieved wim R azteca.  One food  should be used for the standard
       method. However, the decision of what the food should be and the quantity of it are yet
       to be determined.

8.     The test temperature was agreed to be 23 ฑ 2ฐC as most laboratories could accommodate
       that now.

9      The analysis of abiotic factors that may affect test results is needed: this includes issues
       such as the sediment grain size and/or organic carbon content which may affect organism
       survival and/or growth. It was the group consensus that guidance on this issue could be
       developed based upon  existing data  sets developed  in  IL azteca tests with a range  of
       clean .sediments.                  .

                               ;             -                  '-"./.'
The following research needs were identified for IL azteca.

 1     Additional research is  needed to develop approaches to produce known age animals for
       testing.  Alternatively, the manuals would state that if cultures were sieved to  obtain test
       organisms, the age of the animals  retained by the sieve must have been previously
       determined in that culture system.                                         ,

 2     Additional research is needed to evaluate  the  sensitivity of the various  strains of IL
       azteca  to assess whether there is significant genetic drift depending on the waters that
       animals were obtained from  and/or the length of time they have been in monocultures.

 3.    Additional research is needed to develop the standard food for the tests that will provide
        minimal organic carbon input while providing sufficient nutrition.

 4     Additional research on the age  of the test animals  that can be used  and still meet the
        minimum required recovery  is needed.  Testing underway at Duluth may. help to decide
        whether 0-7 d organisms are any different in sensitivity than the 7-14 d old organisms
        which are easier to recover from whole sediment tests.

 5      Additional research is needed on the abiotic factors that may  affect test  results (e.g.,
        organic carbon, particle size).   Data  from Canada  (Burlington), Duluth,  the ARCS
        program, and Mississippi should assist in making the decision.

 6     Additional research is needed to determine the significance of various funding regimes
        on the toxicity test.   Feeding  may alter  the exposure through reduced uptake and
        enhanced elimination  of the contaminants.


                                   v       '27      "-.      "        :  ,.        •..   .     .  •

-------
nhirnnomv* tentans/riparius:

       The workgroup consensus was that the a riparius could be included in the a tenitans.
protocols; these species are fairly similiar.

       Most of the performance-based culture criteria listed for H, azteca are relevant to the
Chironomus spp.  In addition to the chironomids, it is important that the following be considered
in fa& performance-based criteria:

1      Laboratories should measure and record the dry weight of at least one larval stage (e.g.,
    .   day 12) for the purpose of maintaining control charts for each culture. This is a similar
       parameter to young production counts for 1L azteca.

2      Laboratories should record the time to first emergence for each culture and keep this data
 '     .in a control chart.   Fluctuations in time to emergence often precede loss of vigor in
       chironomid cultures.
 3     Laboratories must perform monthly water-only reference toxicant tests  to
       health of their culture organisms or the organisms they purchase from other laboratories.
       The reference toxicant test should be performed as a 96 h water-only test.  Laboratories
       should also evaluate the  slope of their LC50 curve for each reference  toxicant test.
     '  Result of thSe monthly  tests then would be entered into "control" charts; test results
       greater than two standard deviations from the mean LC50 might indicate a cause for
       concern that the cultures are unhealthy.

 4     Laboratories should routinely measure and record the following culture water chemical
       parameters: pH; D.O. ; hardness; alkalinity; and ammonia. Again, it was  suggested that
       this be developed  in manner similar to that used as the control  chart for  reference
        toxicant data.                                                        .        .

 5.     Laboratories should characterize and monitor the quality of the food they use in terms
        of nutrient content and contamination.

 6      Laboratories  should keep records of their culture restart interval and  keep accurate
        records on the age of the brood animals and, as far as. possible, track the source of test
        animals to the age of the brood animals.
  7.     Laboratories should develop standard operating procedures to ensure that
     '   are  renewed and monitored on  a regular  and standardized  manner.
        parameters  such  as lipid content also  might be considered  as a culture parameter to
        determine the health of the organisms.
                                            28

-------
follows:

       The age of the test animals should be: 8 to 12 d old far d tentans and 6-8 d old for
1.
    .  rioanus.
2.
       course of the test.
                 af4?0 Hซ more research wouW be needed before.flus could be added as

       'a criteria for test acceptability.














         done with existing data.


  6      Although various size test chambers are used ,it appearedI that numerous
                                                                  would be acceptable.



   "
         to be determined.                                                        ,

   8.     The test temperature was agreed 'to be 23 ฑ 2ฐC as most laboratories could accommodate
         that now.
                                             29

-------
 9.     The analysis of abiotic factors that may affect test results is needed. This includes issues
       such as the sediment grain size and/or organic carbon content which may affect organism
       survival and/or growth.  It was the group consensus that guidance on this issue could be
       developed based upon existing data sets developed in C. tentans tests with a range of
       clean sediments.
The following research needs were identified for Chironomus spp. toxicity tests as well as those
for the amphipod test above.

I.     Additional research is needed on the abiotic factors that may affect test results  (e.g.,
       organic carbon, particle size).  Data from Canada (Burlington), Duluth,  the ARCS
       program, and Mississippi should assist in making the decision.

2.     Additional research is needed to develop approaches to produce known age animals for
       testing. Alternatively, the manuals would state that if cultures were sieved to obtain test
       organisms, the age of the animals retained by the sieve must have been  previously
       determined in that culture system.

3.     Additional research is needed to develop the relative sensitivity data of midges and other
       taxa.

4.   ,  Additional research is needed to develop the standard food for the tests that will provide
       minimal organic carbon input while providing sufficient nutrition.

5.     Additional research on the age of the test  animals that can  be  used and still meet the
       minimum required recovery is needed.
                                           30

-------
Tjgnhriculus vnrieeatus:

       Many-of te performance-based criteria described above for H, งzteca are relevant to
variegatus.  Additional culture considerations follow:
1
       Laboratories must perform monthly water-only reference toxicant tests to assess  the
       ^S to cultur^organisms or the organisms
       preference toxicant test should be performed as a 9  h water-only test.  ™™*ซป*
       .hould also evaluate the slope  of their LC50 curve for each reference toxicant test
       S* o  the^tSy tes?then would be entered into "control" charts; test resuUs
       gSte* San two  standard deviations from the mean LC50 might indicate a cause for
       concern that the cultures are unhealthy.            '
 2     Laboratories should monitor and record the frequency .wifc which the population is
 ••'     doubling. Again, this might be done using the control chart concept.

 3     Physiological parameters such as lipid content also might be considered as a culture
       parameter to determine the health of the organisms.              ^
 4      Laboratories should routinely measure and record the followin g-cultur e ^ter ch^cal
        dieters- pH- D O.; hardness; alkalinity; and ammonia. Again it was suggested that
        Ke devdopW  in manner similar to that used  as the control  chart for reference
        toxicant data.
 5      Laboratories should characterize and monitor the quality of food i thq
        nutrient content and contamination, particularly for the compounds to be evaluated for
        bioaccumulation .

 6.     Laboratories should keep records of their culture restart interval.

 7     Laboratories should develop standard operating procedures to ensure that their cultures
        are renewed and monitored on a regular and standardized manner.
  above:

  1.
         Given that ^performance-based culture criteria will be partof the requirements for Ae
        hSTtest perfoVmice, the major consensus items for the test protoo* I ซ ซ ซ^
            the vSous parameters of importance for the test method as discussed for H, azteca
         For the time being, 28 d tests are recommended; however, ongoing research may indicate
         SS shorter tests could be used. The use of the performance-based culture cntena for
         SLres wUl be used to judge the health of the animals in the bioaccumulation test along
         3 ttie acceptability criteria.  The test acceptability  criteria will include acceptable
         water chemistry parameters over the course of the test.
                                             31

-------
2.     During tests, the organisms will not have any food added to the test chambers.

3.     The desirable chamber size is more variable than for the previous two species and is
       dependent upon the organism/sediment carbon ratios needed.

4.     The number of organisms used per replicate will depend on obtaining an adequate tissue
       mass for analysis of compounds of concern.  Also, it was suggested  that the ratio of
       organism carbon/sediment organic carbon in the test systems should be on the order of
       greater than 1/10 to 1/100.

5.     The, renewal  of the overlying  water in the sediment test  at a  rate of 1.25  to 4
       volumes/day was agreed upon. The specific procedures were not specified for the waiter
       renewals; however, the intervals should be evenly spaced over 24 h.

6.     The test temperature  was agreed  to be  23 ฑ  2ฐC  as  most  laboratories  could
       accommodate that now.                                ,

7.     The analysis of abiotic factors that may affect test results is needed.  This includes issues
       such as the sediment grain size and/or organic carbon content.
The following research needs were identified for L.. variegatus.

J.     Additional research to determine  the minimum and  optimal  time of exposure for
       bioaccumulative compounds  is needed.  This would include tests that would evaluate
       whether a shorter test (7-10 d) might be possible rather than the 28 d test.

2.     Additional research  to evaluate  gut purging  times  in  relation  to elimination of
       contaminants is needed.  Evaluate  the appropriate time to allow for gut purging and
       whether this is contingent on chemical class.

5.     Additional research to evaluate kinetics of the uptake/depuration for chemicals over a
       wide range of Kow's.
                                         fc
4.     Additional research is needed to evaluate optimal organism carbon/sediment ratios for
       tests.

5.     Additional research is needed to determine the potential of "sediment avoidance" on
       bioaccumulation of contaminants by L.. variegatus.

6.     Additional research is needed on organism loading and doubling of populations during
       kinetic studies.
                                          32

-------
Dav Three

Conclusions and Next Steps
       Dr  Southland opened the session.  She stated that, on the basis of discussion held at
this woShop EPA^ffl develop standard test protocols for acute toxicity and bioaccumulation













 leading to completion of a standard method.

       Dr  Ankley described the results of the freshwater breakout session.  He stated that the
 bases for ^Dedb selection for standardization were: current and historical acceptance; logistical



 %*%%ฃ** organisms are generally epibeathic and matin, testjฃP"™ฃ^*g



 o^re criteria for  H  azteca.  Consensus from the breakout session was reached on what
 criteria mTbe^orZde^d  what criteria should be considered.   Factors that mu^ be
 cSerSlre:  reference toxicants for short-term water ^.^^&^f^^^v










              -  describes the tasks to be performed in developing the final protocols.
                                            33

-------
       Dr. AnMey also summarized major issues discussed concerning the freshwater toxicity
tests.  The following major issues related to the use of H. azteca were covered in the session:

1) Age of test animals.
                                                                                    i
       There appears to be a range of ages most appropriate for testing. Organisms of age 0-14
days are most appropriate.  It may be best to use animals of age 7-14 days, but difficulties in
recovering the animals may be encountered.

2) Length of test.

       The length of the test agreed upon was 10 days with survival as the endpoint.

3) Feeding.

       A minimal amount of food is necessary, but more data on feeding must be generated.

4) Water renewal.

       Limited renewal of test water was recommended (1 volume/day).
                                                                        i         '
5) Sediment volumes.

       Sediment volume in the 1/2 liter range was recommended but it seems acceptable to use
       smaller test volumes.

6) Grain  size.

       There does not seem to be a grain size effect in the short term Hyalella test.

7) Strains of animals.

       There are different strains of Hyalella used for testing. Reference toxicant comparisons
       of the strains are needed.
       In discussing  the  Chironomus  test it was  agreed  that  minimal feeding would be
appropriate (the acceptability of a change in total sediment  organic carbon during the test of
.01% - 1.0% was discussed). Workshop participants did not believe that genetic diversity was
an important issue for Chironomus.
                                          34

-------
       In discussion of the Lum^a^ bioaccumulation test^a num
reached:
1)     The protocol should recommend a 28 day test.
Ti     No feeding is needed during the test.                     ..•,
3      The ScL be conducted with water renewal or static conditions.


I
       necessary.

















  Tpfkin^^^




   are described in the breakout session notes.
                                              35

-------
       Dr. Lee summarized the discussion of the marine bioaccumulation  tests.   Several
approaches to bioaccumulation testing were discussed.  These included:  use of equilibrium
partitioning models; kinetic modeling; and the direct approach of testing field collected sediment.
The development of tissue residue criteria for the protection of human health was identified as
a research need.  It was agreed in the breakout session that the existing 28 day protocols for
Series and Macoma would be standardized. Field validation and round robin testing will be
required to complete development of this protocol.  Over a longer period of time, it will be
necessary to  develop  kinetic models.  It will  also be necessary to  develop guidance on the
ecological significance of tissue residue levels.

       Dr. Southerland concluded the meeting by thanking all the participants for their hard
work and reminded everyone  that methods protocols would be completed for the species
discussed by each workgroup by the end of fiscal year  1993 (October 1, 1993).  The following
fiscal year, if more funds become available, the focus will be on standard chronic test methods
and sediment toxicity identification evaluations.
                                           36

-------
WORKPLANS
     37

-------
38

-------
       The following workplans describe research that will.be undertaken to develop sediment
toxicity test protocols for marine, estuarine, and freshwater organisms.  Bunding has been
proved to complete work on test protocols for species included in the.workplans dunng fiscd
year 1993  Work to develop additional test protocols will be completed if additional funding is
available. The workplans were developed on the basis of discussions held at this workshop.
                                            39

-------
40

-------
                 Development of Sediment Toxicity Bioassay for
                         Marine and Estuarine Organisms
                                  Technical Approach

       This workplan was developed on the basis of discussions held  at an EPA Office of
 Science and Technology (OST)/Office of Research and Development (ORD) sponsored workshop
 on the standardization of sediment toxicity tests. Meetings were also held with ORD scientists
 from the Environmental  Research Laboratories  in Narragansett, Rhode  Island, and Newport,
 Oregon, to discuss and identify the objectives of the proposed  work. In addition, some
 preliminary analyses of the statistical characteristics of acute sediment toxicity data were
 conducted.

      • This workplan describes research to be conducted to standardize acute, chronic,  and
 bioaccumulation tests for sediment  toxicity.  The standardization of these methods is critical
 because of the need to instill consistency in their application in Federal, Regional ,and State
 sediment characterization  programs. This  consistency is  of particular importance to the
 implementation of EPA's Contaminated Sediment Management Strategy.; The strategy calls for
 completion of a national inventory of contaminated sites and continued monitoring to assess the,
 extent and severity of the problem.  These actions will require a nationally consistent means for
 determining sediment quality.

       .The general approach to standardization of sediment bioassays is to start with reasonably
 well-defined procedures, such as the ten-day sediment bioassay with the amphipod Rhepoxynius
 abroriius. and proceed to more complicated, less well-defined procedures, for example, chronic
 .tests and bioaccumulation tests. Field validation of test procedures completes the  cycle of
 standardization prior to implementation of the methods.

        The research described in this work plan supports the development of standard methods
 for acute  and chronic bioassays and bioaccumulation tests.  This  work will concentrate on
 .development  to  the ASTM "Standard Test Method" phase of me ten-day acute  bioassay
 procedure with four species of amphipods.  Other activities will support the development of the
 chronic bioassay and the bioaccumulation test to the ASTM "Standard Guide" phase. The latter
 documents can benefit from the ASTM review process, without the rigorous scrutiny necessary
 for standard methods.

        Standard protocols for acute sediment toxicity tests using the benthic marine and estuarine
 amphipods Ampelisca abdita. Rhepoxvnius abronius. Leptocheirus plumulosus, and Eohaustorius
 estuarius will be prepared. The protocols will include details on: culture and/or acquisition of
 test organisms;  test design; QA/QC requirements;  effects  of abiotic factors;  contaminant
 interactions; reference sediment requirements; relative sensitivity; biological significance of acute
 toxicity; and interpretive  guidance.  Additionally, protocols  for the conduct of the 28-day
 bioaccumulation test,  and for the collection, handling, and spiking of test  sediments will be
 prepared.                                                         ,

        The approach to completion of each research task is presented below.  Research will be
. conducted at the SAIC, Inc. in Narragansett,  RI, testing  center in  consultation with EPA's
 Environmental  Research  Laboratories in  Narragansett,  RI, and Newport, OR,  the  EPA
 Headquarters Office of Science and Technology, and EPA's Tiered Testing  Workgroup.

 •''..'.     .-   -  •       ^           41              ,'         -.''••_'

-------
                 Specific Research Tasks and Methodological Approach
Research to Support the Acute Standard Method:

       There are a number of technical areas where data are needed to support an understanding
of the responses of the four amphipod species to sediment contaminants. Some of the factors that
affect an organism's response to contaminants are temperature, salinity, and grain size. Other
sediment characteristics not related to contaminants, e.g., ammonia, sulfides, and low dissolved
oxygen, may cause toxicity thus confounding the interpretation of contaminant effects.  This
workplan focuses on elaborating test species responses to salinity, ammonia, and grain size
where existing data are lacking. Temperature effects are not important since a standard species-
specific temperature within the upper  tolerance range is normally chosen.  Similarly, low
dissolved oxygen or sulfide toxicity will not be addressed because the test chambers are well
aerated during sediment exposures.

       There also are no data on the  relative sensitivity among these species to dissolved and
sediment-associated contaminants. Field validation data also are  lacking for three of the four
species. In addition to the collection of supplementary data through the conduct of laboratory
experiments, existing data on these elements of the tests"will be gathered and synthesized.

    "   The experimental design addressing each of the issues discussed below will be prepzired
and agreed upon in consultation with technical experts from the ORD's Environmental Research
Laboratories and EPA's Office of Science and Technology.

Grain Size  Tolerance:
                                                                 , \
       Most marine and estuarine organisms prefer a particular range of sediment type that may
be  related to feeding or burrowing habits. In contaminant effects studies, sensitivity to particle
size is important since unusual stress associated with extremes of the particle size range may
induce mortality. The effect of fine particle size as an artifact causing non-contaminant induced
mortality in Rhepoxvnius abronius is well understood, and algorithms describing that relationship
have been developed.  It is possible that the opposite effect, i.e., coarse particle size-induced
mortality, may occur in Ampelisca abdita. While contaminants are generally associated with fine
sediments, contaminated coarse sediments can be extremely toxic also. Experiments will  be
conducted to identify the potential for grain size effects in both Ampelisca and Leptocheinjs.

       The hypothesis to be tested is  that extremes in sediment particle size do  not cause
excessive mortality in the  test organisms. To test this hypothesis, a range  of particle size
treatments (up to five) will be established by mixing,- by volume, non-toxic coarse sediments
with silt/clay sediments. Natural sediments with different particle sizes may also be tested: I&ch
species will be exposed to these sediment mixtures using the existing ASTM procedure for the
conduct of 10-day solid phase bioassays.
                                           42

-------
not exist at present.            .    .  -                  -



Ammonia Tolerance:






                      ,0 the other three test species in 10-day *ttc .exposures.
            sets of expend              wu




 sediment-spiked ammonia.


 ^r^ "^^
 ammonia concentrations will be determined.


 Salinity Tolerance:




  ugss^m and An^.^^^-^m^SfaSL Detest salinity
  ^faSpCs^U rKCSry SwUl be evaluafcd in Ught of existing info^on
  on the distribution of these two' species relative to salinity.

        This research will not address the potential interactive affects of sublethal salinity stress
  and contaminant toxicity within the non-lethal salinity range.
                                         43

-------
 Relative Sensitivity:

       Typically, only one of the four amphipod  test species will be used in a solid phase
 assessment of sediment toxicity.  The species  of choice would be selected  for a number of
 reasons  including:  availability;  regional  interest;  and  particle  size  compatibility.  The
 interpretation of a toxic response will  require an understanding of the sensitivity of the test
 organism relative to that of "benchmark" species for which a large data base exists. Large data
 bases do exist for Rhepoxynius and Ampelisca.  however, direct comparisons of the two species
 are rare.                                                                  .
       Experiments will be conducted to test the hypothesis that there are no differences in
 sensitivity of the four amphipod  species.  The relative sensitivity to two  inorganic and two
 organic compounds will be determined through two sets of experiments with the four species:
 four-day water-only and 10-day spiked sediment bioassays. LCSOs for each species and exposure
 type  will be determined.  The relative sensitivity to field: sediments of known  contaminant
 concentrations  and toxicity will also be determined in round robin testing described below.
 Further information will be gained by analysis of existing data on the relative sensitivity of these
 species.
Field Validation:

       The ultimate utility of these acute bioassay methods will depend on the extent to which
they predict contaminant effects in natural populations and communities. The toxic responsie in
Rhepoxynius has been related to the absence of amphipods and alterations in benthic community
structure.

       A field validation of the acute response is necessary ,for Ampelisca. Eohaustorius. and
Leptocheirus. Data are currently available from the EMAP and NOAA Status and Trends isites
on the benthic community composition and-abundance of amphipods. Concurrent data are also
available on  Ampelisca toxicity for many of .these sites. These data will be reviewed and
analyzed to determine if sufficient data exist to field validate the acute test for this species. The
criterion for validation will be the co-occurrence of toxicity with decreased amphipod abundance
and/or significant benthic community effects.

       Analysis  of these  data  will identify sites that may be used  to validate  the test  for
Leptocheirus. Additional samples, not to exceed 20,  will be collected in cooperation with these
programs for toxicity tests with this species. Similarly, if the data on Ampelisca are insufficient,
this species will  also be tested.
                                          44

-------
Testing Summary:
ฐ?n tmDelisca - 10-day sediment - 5 treatment levels - 2
 o   .uSSSU-lMiy sediment -5 treatment levels -2 runs
                  - 10-day spiked sediment - 5 treatment levels
                   - 10-day spiked sediment - 5 treatment levels
                    ^^ฃ%^-,*
                   - 10-day spiked sediment - 5 treatment levels
Field Validation         '"..,.
     AmEeUsca - 10-day - 20 sediments
     Le^tocheirus - 10-day , 20 sediments
   o
  At least 5 replicates will be used per run.
                                        -1-'
  Research to Support the Chronic Standard Method:

        Research wiU be initiated to dfel^
   j^j^ and Anodto .MJJf^S ^vt'nlnS ResSrcrilboratory in Newport
   development and validation by the E^e^™nm^.h condubted to develop acute, test
   Oregon (ERL-Newport) and others. Some of ftc ^™,^A.g EnvirOnmental Research
   methods wDl apply  to the ^%^^^gisett) is also supporting research on
   Laboratory in Narragansett, ^^^SSh with tbeฃ two species will be
   chronic sediment bioassays ^^^^ S for the ERL-Newport and Narragansett
   defined at a later date when ^.c^m^^s^ith up to five treatments each will be

   ^rtr*^^^^
   of S work at & two EPA E8L laboratones.
                                     45

-------
        Since the ability to culture .test species will be a requirement of chronic methods,
 additional research will address the culture requirements of Ampelisca. Short-term experiments
 will investigate the effects of culture container size and configuration, and feeding procedures
 and frequency on the size and fecundity of laboratory populations.


 Research to Support the Bioaccumulation Standard Method:

        The accuracy of the marine sediment bioaccumulation test is poorly quantified, and the
 extent to which it predicts tissue residues under field conditions is unknown. The accuracy, as.
 defined by a field validation of the laboratory results, is critical to the future development and
 application of this test. A key to the laboratory to field comparison is the achievement of steady
 state conditions in the laboratory exposures. Also, ideal circumstances would dictate that the site
 chosen be inhabited by the same organisms being used in the bioaccumulation test.

        Research described in this workplan will  provide an initial field validation of the
 bioaccumulation method using Macoma nasuta and  Neries virens.  This research  entails  a
 minimum of a 60-day laboratory  exposure  to  a  naturally  contaminated   sediment  with
 measurements of tissue residues in each  species at a minimum of two time intervals (28 and 60
 days). These values will be compared to those measured in field collected organisms, preferably
 the same species, inhabiting the same sediments.

       Up to 50 tissue residue and eight sediment residue measurements will be required for this
 research. Chemical  analyses  will target high and  low molecular weight PAHs, metals,  and
 chlorinated compounds.

       Criteria for the selection of an appropriate site include the presence of the test organisms,
 or an  adequate sediment-ingesting  surrogate, and  concentrations  of target  contaminants
 sufficiently high to be bioaccumulated, but not high enough to cause mortality in test organisms.
 Only one site validation  will be conducted  due  to the limited availability of resources.
 Therefore, the site will be carefully selected after consultation with ORD and OST experts. The
 same consultation will be required to finalize the experimental design.

         •                                     ',''*•
 Statistical Analysis of Acute Bioassay Data:

       The currently accepted criterion for the assignment of toxicity to a sediment sample relies
 on the detection of a statistically significant decrease  in survival in a test sediment relative to that
 in a control (or reference) sediment. This research will provide a rigorous analysis of acute test
 data  for the four  species  to  determine  a  level of statistical  significance that relies on  the
 variability of the test performance among and between test runs. The goal of these analyses is
 to describe procedures for examining the statistical  nature of acute toxicity data bases  relative
 to variations in control mortality, test precision, and the ability to determine minimum detectable
differences. Preliminary analyses of selected Ampelisca data indicate that minimum detectable
differences may range from 20 to 30%. Ultimately, these statistical differences can be compared
to differences in mortality that are considered biologically significant.
                                           46

-------
 need, if any, for data transformation.
 Acute Round Robin Tests:
-
                                                           K^sra
 conducted to provide direct comparison with the other two species.
       Four laboratories will be evaluated in this task. Up to four naturally contaminated and

  Preparation of Draft Standard Methods:

        Draft standard methods will be prepared for acute sediment, bioassays witii the four
        The ASTM format will be followed for this and all other methods developed ^nderthis
  workpla? ifm^foZt, the documents will be submitted for review and approval under the
  ASTM Subcommittee E47.03 sediment toxicology balloting system.

         A draft euide will be prepared for chronic sediment bioassays using T^ptocheirus. This
   included in the draft guides.

         Two draft guides for the 28-day bioaccumulation test with Macoma and Neries. will be
                               be baled on the existing EPA method for the former spec.es.
   t/1. WL/u-L v*X* • •*• • i*"*-™- ••-——•	               . ^
   That'EPA method also is a draft ASTM guide.


                                       -   47

-------
       Additional standard methods for sediment collection, handling, and spiking will be
prepared in the ASTM format. The sediment spiking procedure will follow those developed by
the ERL-Newport laboratory for the sediment quality criteria program. The sediment collection
and handling document will be based on an existing guide prepared by ASTM. This document
will be revised in conjunction with ongoing efforts to develop standard methods for freshwater
sediment bioassays.  Specifically, a committee of experts will be formed to come to consensus
on key issues. This group will meet at the fall 1992 and spring 1993 ASTM meetings to discuss
the proposed revisions to the existing document.
                                         48   •

-------
                                     Schedule
Acute Testing:
Complete ammonia tolerance tests' with^he^xynius abronius, Le^tocheirus plumulosus, and
Fohaustorius estuarius - data report (Jan/93)
Complete grain size tolerance tests with Ampdisca abdita and L^tocheirus r4umulosus - data
report (Jan/93)
Complete salinity tolerance tests - data report (Jan/93)

Complete comparative sensitivity tests with spiked sediments - data report (Mar/93)

Complete field validation data analysis and testing - data report  (Sept/93)
 Chronic Testing:
 Complete development of culture methods for Ampelisca - draft protocol (Sept/93)


 Bioaccumulation Tests:
 Complete laboratory testing and chemical analysis for field validation - data report (Sept/93)
  Statistical Analysis Procedures:
  Completion of statistical analyses for mirdmum detectable difference for each of the fourspecies
  - data report (Jun/93)
  Acute Protocol Round Robin:

  Completion of acute test round robin - data report (July/93)
                                            49

-------
Document Preparation:




Draft Standard Method for Rhepoxynius abronius Acute Test (Apr/93)



Draft Standard Method for Ampelisca abdita Acute Test (Sept/93)



Draft Standard Method for Leptocheirus plumulosus Acute Test (Sept/93)




Draft Standard Method for Eohaustorius estuarius Acute Test (Sept/93)




Draft Guide for Macoma nasuta Bioaccumulation Test (Sept/93)




Draft Guide for Neries virens Bioaccumulation Test (Sept/93)



Draft Guide for Leptocheirus plumulosus Chronic Test (Sept/93)



Draft Standard Method for Sediment Spiking (Apr/93)




Draft Standard Method' for Sediment Collection, Handling, and Storage (Sept/93)
                                        50

-------
                Development of Sediment Toxicity Bioassays
                         for Freshwater Organisms
                              TM-hnical Approach
      This workplan was <^op^
workshop on standardization of sediment  cmaty ^^^^ ^ |nvironmental
contaminants  in sediments to aquatic ?f"*ฃ^*SS Si areas where water quality
toxicology. Mounting evidence exists of ^^^^^^^^-^b^l^l^
criteria are not exceeded, yet orgamsmsare *^*™^ J^ent ^ost assessments of
placed on evaluating ซPซ*^' f**^               considered a safe repository of
:Sn^
considering the fate of chemicals in sediment.
       A variety of methods have "^
 procedures range in complexity frฐt^ฐ^
 Contaminants on single species     "^S^iiictato whole sediment (often

       Ideany, a sediment test should ^
 study is to screen a large number of^ ซm a^y man     ^^^ ^^^^
 contaminated sediment can^be atxop^^^^S^ with the ability of an animal
 of chemicals in sediments that are not fJ^STo^S chronic effects and long-term
 to develop grow  ซ''*^ J^SSS^^
 bioaccumulation of chemicals from sed ^""f1  ^   coiiminants.  Most estimations of
 and to understand the enyironmen^ •*&*%ฃ baled on T- to^-d exposures with midges,
 . chronic sediment contaminant ^ effects have ^ ^^  .^ u^cyde exposures may not
   from responses to natural sediment characteristics.
                                         51

-------
        Direct comparisons of animals exposed in the laboratory and in the field are required to
 verity results from laboratory testing procedures. The assumption that laboratory results for a
 specific sediment represent effects of similar sediments in the field needs to be evaluated.
 Hazard evaluation of contaminated field sediments that integrate data from laboratory exposures,
 chemical analyses, and in situ field assessments provide strong complementary evidence of the
 degree of pollution-induced degradation to aquatic and  benthic communities.

        The goal of this research project is to develop state-of-the art, standardized protocols for
 assessing the potential effects of contaminated sediments on aquatic ecosystems.   These
 laboratory tests are an essential component to the tiered testing approach currently being
 developed by EPA.  The general strategy behind the research is to start with the standardisation
 of reasonably well-defined test procedures (10-d acute toxicity tests with benthic invertebrates),
 proceeding  to less  well-defined protocols (bioaccumulation tests, food chain models, chronic
 toxicity tests, toxicity  identification evaluation), and ultimately  culminating  in field validation
 of the tests.  Because many contaminants of concern in sediments bioaccumulate, this research
 will emphasize development and validation of toxicity and bioaccumulation tests to residue-effect
 endpoints based  on tissue concentrations. Part of this effort will involve developing toxicokinetic
 and metabolism models for species exposed to different classes of representative  sediment
 contaminants. This information is needed in order to develop realistic models for predicting
 exposure of organisms through the food chain, and will also provide a technical  basis for
 assessing the use of risk based residue-effect models.

       The following objectives (elements) and associated timelines are based  on the assumption
 that support  for this research effort will be available at comparable  levels  for 3 to 5 years.
 Available FY92  and FY93 funding will support only research activities dealing with Objectives
 1, 2,  and 3. Research will be  conducted  at the U.S. Fish and  Wildlife Service National
 Fisheries  Contaminant Research  Center in  Columbia,  Missouri  (NFCR-C)  and  EPA's
 Environmental Research Laboratory in Duluth, Minnesota (ERL-Duluth) in consultation with an
 established workgroup  of experts on freshwater sediment toxicity testing,  EPA's Office of
 Science and Technology, and an EPA Tiered Testing Workgroup.
1.
2.
3.
4.
5.
6.
7.

8.

9.
                       Specific Research Objectives

Standard Protocol for an Acute Toxicity Test with Hyalella azteca (FY93)
Standard Protocol for an Acute Toxicity Test with Chironomus tentans (FY93)
Standard Protocol for a Bioaccumulation Test with Lumbriculus yariegatus (FY931)
Toxicity Identification Evaluation (TIE) Procedures for Contaminated Sediments (FY94)
Standard Protocol for a Chronic Toxicitv Test with Hvalella azteca rFY94)
Standard Protocol for a Chronic Toxicity Test with Chironomus tentans (FY95)
Develop a Generalized Model for-Predicting the Metabolism of Common Sediment-
Associated Contaminants in Benthos and Fish (FY95)
Develop an Effects-Based  Tissue Residue Model for Assessing the Risk of Sediment-
Associated Contaminants (FY96)
Summarize Field Validation Studies for Standardized Toxicity and Bioaccumulation Tests
for Freshwater Sediments (FY96)                         .
                                          52

-------
                  Fvperimentfll Design anf* Methodological Approiach



      Investigations of sediment toxicity and  bioaccumulation are  limited by -k lack : .of
understanding of the factors controlling contaminant availability in sediment. Additionally, a
"act Turitide stendardized methods also Umits the use of sediment tests m ^ntammation
assessments.  ASTM Subcommittee E47. 03 on Sediment Toxicology 'has *^&ฃ*ฃ
assessing the bioavailability of contaminants associated with sediments (e.g. , ASTM E 1383 92
^sSd Guide  for Conducting  Sediment Toxicity Tests witfi Freshwater ™f* ฃ
These guides are used to evaluate the toxicological hazard of contaminated sediment  soil
sSe Sng fluids, and similar materials. The Subcommittee developed general guides and
no? sSndarftest methods or protocols, because  most procedures for *M.coott^
sSinSnt have only been Decently  developed.   Definitive protocols are needed which describe
specific test methods.                             .



Objective 1:  Standard Protocol for an Acute  Toxicity Test with Hyjdella gzteca (FY93)
           •  i         • '   ^.         •        '      •,  • •
      ' The protocol will include details for culturing and testing the amphipod,  including test
 svstem design   Also covered "will be the development of a standard reference sediment,
 Sures for'reference toxicants, procedures for interpreting the effects of abiotic factors (e.g
 narticfe sSe)^ or ^results, use of cell lines for screening complex hydrophobic compounds and
 deteS me^tency of these .compounds to aquatic organisms, the results of a relative
          Lalys^Cfrom a series of single chemical tests) for K azteca, evaluation of genetic
           Moratory cultures of the amphipod, and results of preliminary round-robm studxes.
   ,     Hvalella azteca will be cultured to produce known-age or known-size animals. MFC^-C


 of cukures wffl be compared using known-age or mixed-age methods and various diets.
        A standard control sediment will be developed for use in determining
                                                                                   and
      f\ dlxUlUAiU vWil U.WA tjw^*."*"-'*" ••	^     J.         .       .  .   *
nf the test and will facilitate inter-laboratory comparisons.  A particle size
con* nuSoTof total organic carbon will be selected  to  be  representative^ freshwater
SeSsNTCR-C will evaluate KC1 and ERL-Duluth wiU evaluate CuSO4 for use as a

reference toxicant.

       NFCR-C wiU establish a culture of the ERL-Duluth strain of HyjMla a^a.  Relative
sensitiv^of tMs^n will be compared to the NFCR-C strain. Taxonomy of both strains will
be confirmed by an identified expert.
                                           53

-------
       Preliminary round robin studies will be conducted by 8 laboratories using water only exposures
 (Phase 1) and whole sediment exposures (Phase 2):
       a.

       b.


       c.
Phase 1: Water only, 4-d exposure, 3 species (e.g., common strain), 1 reference
toxicant (KC1).
Phase 2: Whole sediment, 10-d toxicity exposures,  3 species (e.g., common
strain),  2 sediments (medium and high toxicity) + control. Versions of ERL-
Duluth methods would be  used.
Timeline:                             .
1. Identify methods and laboratories for testing
2. Phase 1 testing: October,  1992
3. Phase 2 testing: February, 1993
Objective 2:  Standard Protocol for an Acute Toxicity Test with Chironomus tentans (FYS>3)

       The protocol will include details for culturing and testing ฃL tentans. including test system
design.  Also covered will be  the development of, a standard  reference sediment,  procedures for
.reference toxicants, procedures for interpreting the effects of abiotic factors (e.g., particle size) on test
results, use of cell lines for screening complex hydrophobic compounds and determining the potency
of these compounds to aquatic organisms, the results of a relative sensitivity analysis (from a series of
single chemical tests) for C. tentans. and results of preliminary round robin studies. Preliminary round
robin studies will be conducted by 8 laboratories using water only exposures  (Phase 1) and whole
sediment exposures (Phase 2).                                      ,           ,
Objective 3: Standard Protocol for a Bioaccumulation Test with Lumbriculus variegatus (FY93)

       The oligochaete Lumbriculus variegatus is the most promising benthic test species available for
a standardized  freshwater  bioaccumulation test.   Research will  be focused on field validation of
bioaccumulation tests with Lumbriculus. as well as analysis of the kinetics of bioaccumulation of
different classes of chemicals of concern. Lumbriculus variegatus will be cultured at NFCR-C. ERL-
Duluth will supply their SOPs and animals.  NFCR-C will determine performance of the cultures using
reference toxicants and estimates of population dynamics.
                                              54

-------
 Specific Research Tasks to Develop Acute Tnyicitv and Bioaccumulation Test Protocols

      Specific  research tasks that must be completed to develop the acute toxicity and
bioaccumulation test protocols are identified below. Each area of research is identified with a
group or groups having primary responsibility for conducting the work: (Columbia = NFCR-C,
WSU = Wright State University, Athens  = NFCR-C Athens Georgia Field Research Station).
An area of research designated "round robin" will be evaluated by laboratories participating in
the round robin tests.  An "*" indicates areas of research that will not be completely resolved
bv the end of fiscal year 1993.  Best professional judgment will  be used in some instances to
make decisions regarding  areas of research indicated with an ''*V  An area of research
designated with an "@" was identified as a high priority research need at EPA s test method
standardization  workshop.   NFCR-C will  be responsible  for  writing the final document
describing the standard protocols. Chemical analyses will be done by the U.S. Fish and Wildlife
Service Patuxent Analytical Control Facility.  This laboratory will meet QA/QC requirements
and anticipate a 30 to 90 day turn around time for samples.

1.    CULTURING

       a.1     amphipods: known-age vs. mixed cultures  .

               1.    evaluate sensitivity to suite of compounds with different modes of action
                    (Columbia, Athens
              2.    consistency in size of organisms between known-age and mixed cultures
                    (Columbia, WSU, @)
                                         •ป•-.'•           '
       b.     use of reconstituted water (*)

       c.     diet(*)

       d.     performance criteria  (WSU, *, @)

 2,    WHOLE SEDIMENT TESTING

       a.     temperature: 20 to 25 ปC (*)

       b.      static renewal 1-4 volume additions/day (*)

        c.      methods for static renewal:  evaluate water quality with various static renewal
               exposure systems (All)                    ,

        d      chamber size: 30 mL to 1 L (typically 250 to 300 mL); volume of sediment: 100
               mL (minimum for 300 mL chamber);  sediment to water ratio between 1:1 and
       ,        1:4 (round robin, @)

  '   •  •      • •      .-.•'-       .-        55       -        -  •   . .      •-;...'.

-------
e.     known-age animals:

       1.    Hyalella azteca: 0 to 14 d old

             a.     evaluate using reference toxicants (All, @)

             b.     behavior in sediment by life stage (WSU, Columbia)
                                           \
       2.    Chironomus tentans: 10 d old (*)

             a.     evaluate using reference toxicants (All, @)

       3.  .  Lumbriculus yariegatus: adults (*)

             a.     evaluate using reference toxicants (All, @)

       number of animals/chamber: minimum 10 (*)

       number of replicates (chambers): power analyses are needed to determine desired
       number of replicates (Athens, @)

h.     feeding (All, round robin, *, @):

       1.    Hyalella azteca: 6 mg rabbit pellets/MWF/20 animals (Columbia); 0.8 mg
              YTC/d (EPA); methods will be compared

       2.     Chironomus tentans: 4 mg Tetramin/d/10 animals

       3.     Lumbriculus variegatus:  no feeding for bioaccumulation testing, 20 mg
              trout starter every 5 d/10 animals in toxicity testing

i.     water quality: evaluate use of reconstituted water (*)   t

j.     photoperiod: 16:8, minimum 25 foot candles (*)

k.     endpoints: survival, growth (round robin, @)            ,

1.     test acceptability criterion (round robin, @).
f.

g-
                                    56 '

-------
3.    WATER-ONLY TESTING

      a      Reference toxicants: Water only, 4-d exposures, performed monthly  Columbia
             wm Sate KC1, WSU will evaluate CdCl2, and ERL-Duluth will evaluate
             CuSO4

      b.     Use of reconstituted water (All, @)

      c.     Use of phenol or another non-ionic organic as a reference toxicant for water-only

             or sediment testing (*)      '                           ,

 4.    STANDARD CONTROL SEDIMENT (@, Columbia, Athens)


 5.    PROCEDURES FOR  EOTERPRETTNG EFFECTS  OF  ABIOTIC  FACTORS

       (Duluth, Columbia, WSU, .<ง>)

       Reconstituted sediment will be used to evaluate particle size and organic carbon. . ERL-
       ShT^veSS databases and regression equations with 50 sediment samples for
       2 "™dTEW.Dulซth is developing manuscripts dealing wi* interpreting the effects
      •of ^Sa. WSU is evaluating the influence of low dissolved oxygen.


 6.    RELATIVE SENSHTVITY ANALYSIS (Duluth, @)


       ERL-Duluth is developing a database for 12 chemicals with all 3 species using 10-d .flow-  ,

       through water-only exposures.    .                                     '


 7.    GENETIC VARIABILITY OF Hyajella azteca (AU, @)

       miumbia and WSU have started cultures of the ERL-Duluth strain of HyaMla aztecl.
       "^™ofsซ*ns to a suite of compounds with different modes of action will

       be evaluated.

  8.     PRELIMINARY ROUND-ROBIN STUDIES (WSU, @)


        a.    Toxicity testing: 8 laboratories

  ^            1     Phase 1: Water only, 4-d exposures,
                     next spring), KC1 reference toxicant. October


               2     Phase 2: Whole sediment, 10-d toxicity exposures, 2 s^iments + control.
                     Versions of ERL-Duluth methods used. February 1993.
                                         57

-------
9.    EXPERIMENTAL DESIGN FOR FIELD VALIDATION OF BIOACCTJMULATION
      TESTS WITH LTJMBRICULTJS

      a.     Objectives


             1.    Kinetics of bioaccumulation of different classes of chemicals of concern
                   with Lumbriculus


             2.    Field   validation  of  laboratory   bioaccumulation  exposures  with
                   Lumbriculus

      b.     Experimental design
                                 i                           \         -

             1.     Field-collected sediments (2 sediments)


                   (Select sediments with broad, range of K^ compounds (up to K^ log 7-8);
                   organic carbon/lipid normalize.


                   Suggested location for sediment collection:


                   a.    Little Scioto River in Ohio: high PAHs and metals, possibly PCBs


                   b.    Huntsville Alabama:  DDT and metabolites

            2.     Field-collected oligochaetes (5 samples/sediment)


            3.     Laboratory-exposed oligochaetes  (56-day exposure sample over time)

     c.      Field collection                           '


            1.    Field-collected oligochaetes ,


                  5 samples x 2 sediments =  10 oligochaete samples

            2.     Field-collected sediment


                  A.    2 sediments: Little Scioto and 1 Huntsville


                  B.    Collect multiple grabs of sediment (about 4 L/grab).  Homogenize
                        and split into two subsamples-.  one 2-L subsample  for sediment
                        chemistry  and  a  second  2-L  subsample  for  field-collected
                        oligochaetes.   Sieve oligochaetes in the field.  Repeat 4 L grab
                        sampling until enough biomass and sediment is collected.  Ship
                        oligochaetes and sediments to Columbia.  No depuration of field-
                        collected oligochaetes  (comparison  to  28-D  sample  w/o
                    1    elimination).
                                      58

-------
 d.      Chemistry samples (metals and organics sampled from the same replicate)

        1.    Chemical-specific analyses:

              A.    Little Scioto River: PAHs, metals, possibly PCBs

              B.    Huntsville: DDT and metabolites

        2.    Sediment

              2 replicates/sampling period
       •   !    x 3 sample periods (day 0, 28, 56)
               = 6 samples/sediment

        ,3.     Field-collected oligochaetes                            '

               5 replicates/sediment

         4.    Laboratory-exposed oligochaetes

               A.    Uptake: day 0, 2, 4, 7, 14, 28., 56

                      1.     3 replicates/sampling period                  .
                             x 6 sampling periods (days 0, 2, 4, 7, 14, 5t)
'  '    ,                       =18 samples/sediment

                      2.     5 replicates/sampling period
                             x 1 sampling period (day 28)             ;
                            '.= 5 samples/sediment               ,
    /                                         .
   >                    3     Total: 23 oligochaete samples/each sediinent

                B.     Elimination: hour 12, 24, 48, 72, and day 7

                       1.     2 replicates/sampling period        ,
                              x 5 sampling periods
                              x 2 treatments (with or without sediment)
                              =  20 samples/sediment
                                       59

-------
                       Additional Research Proposed for FY94-96
 Objective 4:  Toxicity  Identification Evaluation  (TEE) Procedures for  Contaminated
               Sediments (FY94)

        A draft document describing preliminary methods for sediment TEE has already been
 completed; further resources will enable completion of research focused upon issues such as pore
 water preparation, isolation and fractionation  of high  log K™ non-ionic organics,  species
 selection, TDB on whole  sediments, and field validation.
 Objective 5:   Standard Protocol for a Chronic Toxicity Test with Hvalella azteca (FY94)

       In  addition to  the research issues identified under Objective  1, a key  factor to be
 addressed  in the development of a chronic test would focus on appropriate toxicity endpoints.
 In addition, the length of the test will be evaluated; it may be, for example, that little additional
 information is gained in long-term tests as compared to short-term tests.
 Objective 6:  Standard Protocol for a Chronic Toxicity Test with Chironomus tentans
              (FY95)

       See Objective 5.
 Objective 7:  Develop a Generalized Model for Predicting the Metabolism of Common
              Sediment-Associated Contaminants in Benthos and Fish (FY95)

       Residue-based risk assessments for hydrophobic sediment-associated contaminants are
 highly dependent on accurate prediction and measurements of bioaccumulation.   For super-
 hydrophobic chemicals, bioaccumulation potential can be significantly overestimated by models,
 which assume no metabolism, or by empirical exposure that are of insufficient length to account
 for the kinetics of biotransformation.  Procedures will be developed whereby computer-assisted
 predictions of metabolic rates  can be used to  refine bioaccumulation estimates and identify
 associated uncertainties.
Objective 8:  Develop an Effects-Based Tissue Residue Model for Assessing the Risk of
              Sediment-Associated Contaminants (FY96)

       Current standard  guides  must be  modified  to provide a residue-effect based  risk
assessment approach for very hydrophobic chemicals.  Studies must be longer in length Jind
incorporate  both dietary  and water column routes  of exposure  to adequately  quantify
bioaccumulation.   Toxicity endpoints  must also include  effects  on  reproduction  and the
probability of these effects must be related to residue levels.  Research will be directed to
provide improved test protocols and predictive models for assessing risk.
                                          60

-------
               SrhPdnle for rnmpletion of Pmiliicts in Fiscal Year 1993

      The final product of workifunded in FY93 will be standard protocols for acute^oxicity
      The ASTM format will be followed for all methods developed under this ™^; Jฐ
this fomat,  ^d^ents>m be submitted  for review  and approval  under  the ASTM
Subcommittee E47.03 sediment toxicology balloting system.
                                           61

-------
62

-------
      OUTLINES
         OF
SPEAKER PRESENTATIONS
          63

-------
64

-------
EPAys Contaminated Sediment Management Strategy
Elizabeth Southerland,  U. S: EPA Office of Science and Technology

       In the 1980s EPA documented the extent and severity of contaminated sediment problems
at sites throughout the U.S.  Concerned with the mounting evidence of ecological and human
health effects, EPA's Office of Water organized a Sediment Steering Committee chaired by the
Assistant Administrator of Water and composed of senior managers in all the EPA offices with
authority to handle contaminated sediments and EPA's ten Regional offices'.:

       Over the past two years this committee has beenpreparing an Agencywide Contaminated
Sediment  Management Strategy to coordinate and focus EPA's resources on contaminated
sediment problems. A draft outline of this strategy was released to the public this year to serve
as a proposal for discussion in three national forums scheduled  for April, May, and June. The
draft  strategy is designed around three major principles:

       1.      In-place sediment should be protected from  contamination to ensure that the
              beneficial uses  of  the nation's  surface  waters  are maintained for  future
              generations;

       2.     Protection of in-place sediment should be achieved through pollution prevention
              and source controls;

       3.     Natural recovery is the preferred remedial technique.   In-place  sediment
              remediation, will be limited to high risk sites where natural recovery will not
              occur in an acceptable time period and where the cleanup process will not cause
              greater problems than leaving the site alone.

       The draft  strategy includes several component strategies:  assessment, prevention,
 remediation, dredged material management, research,  and outreach. A brief summary of each
 of these elements  follows.                                      -

       In,the assessment strategy EPA is committing to  develop a national  inventory of
 contaminated sediment sites and a pilot inventory of potential sources of sediment contamination,
 based on existing data. The two types of inventories will be complementary because the source
 database can be used to predict where sediments are contaminated in unsampled areas.  The
 inventories will be designed so that EPA's prevention and remediation programs can use them
 to focus their resources on cleaning up the top priority sites and sources.  Another key element
 of the assessment  strategy is the commitment to develop a consistent; tiered testing strategy that
 will include a minimum set of sediment chemical criteria,  bioassays, and bioaccumulation tests
 that all programs  will agree to use in determining if sediment are contaminated.

        The prevention strategy includes a variety of pollution  prevention measures and source
 controls. The scale of contamination will guide the choice of a particular set of these measures.
 If a  sediment contaminant is causing harm or risk at numerous sites nationwide, it may be

                                           65*  - ''  '   '    '•.-  '..'..,':/.

-------
 relatively inefficient to deal with the problem on a site-by-site basis.   Instead,-the strategy
 discusses nationally applicable responses, such as prohibitions or use restrictions under TSCA
 or EOFRA, technology-based effluent limitations for industrial dischargers, or a national initiative
 to revise water-quality based limits in NPDES permits.  If atmospheric deposition appears to be
 a primary source of contamination, responses under the Clean Air Act will be considered.
 Where sediment contamination is a concern at particular sites, but not on a national scale, case-
 by-case assessments and response actions are recommended. Based on narrative and chemical-
 specific criteria and standards, EPA or a State can develop NPDES permit limits for discharges
 from'industrial sources, municipal sewage treatment plants, stormwater outfalls, and combined
 sewer overflows.  States that have ndnpoint source control programs can take actions to reduce
 the contributions of these sources to sediment contamination.

        EPA may, remediate sediments  under CERCLA, RCRA, CWA, and TSCA.   The
 remediation programs will use the national inventory to assist in selecting sites for cleanup and
 the consistent tiered testing to  assist in identifying contaminated areas and establishing cleanup
 goals.  The remediation strategy emphasizes that sources of contamination should be controlled
 prior  to remediation  efforts  unless the contaminated sediments pose a  sufficiently  great
 environmental hazard.   In making remediation decisions, the strategy also points out that it is
 important to consider whether contaminated sediments at a site can be transported to downstream
 or offshore areas if left in place, thereby increasing the size of the contaminated area and making
 future remediation efforts much more difficult.  Other factors to consider include the timefraiwe
 for natural recovery, the potential for contaminant mobilization during remediation,  and the
 feasibility and cost of various treatment and removal options.

       The  maintenance of our nation's waterways for navigation requires  the dredging and
 disposal of 250 to 450 million cubic yards of material each year.   Dredged material testing
 manuals prepared jointly by EPA and the Corps  of Engineers recommend  the chemical and
 biological tests that should be conducted to determine if the material is contaminated and must
 be disposed of using special procedures. The tests selected for the Agencywide contaminated
 sediment strategy will be included in these dredged material testing manuals.  The strategy also
 outlines additional guidance that will be developed by EPA  and the Corps to improve the
 management of these materials.         . ,                                 >

       The  research strategy  outlines all  the work that EPA's  Office of Research  and
 Development  (ORD)  has planned  on sediment  chemical  criteria,  sediment  bioassay  and
 bioaccumulation test, fate and transport models, and remedial techniques.  ORD is establishing
 a Resource Center to provide  EPA  offices  with centralized technical assistance in evaluating
 sediment contamination and will also sponsor workshops and training sessions throughout the
 country.

       The outreach strategy describes how EPA will work with other Federal agencies and State
 agencies to  coordinate  EPA's  contaminated sediment activities with  their efforts.  EPA will
 strive to ensure that these agencies share  sediment related research findings and innovative
technologies.  In  addition, EPA is proposing  a two-way public awareness program that will
disseminate contaminated sediment information to  the public and also incorporate information
from the public into EPA activities.                    '      '
                                          66

-------
67

-------
cซ
J
<
O
O

        z.
        ^ ^H



!s -
TS ^
^^ ^^

Is

tf' ^
     O

     = ^
     ซ. c
     C5
     >; C

    = ซ
^ 4)    -—
       W)
       Eซป
        5
     5 S
         Dฃ
     Z O "C
     tn --
    K   Q
            68

-------
69

-------
 S
 o
CL,
         en
         en
         C
         C5

         •••

         'S
ent
i
sess
                 c
                 c
         en



        "en

        S
en

i  i
O  83

y  M
S  ฃ
   e.
                  en

                  li
         S

         03
    O
S W
 2  ซ

 ""*  s-

 e
 0)

.i  B
T5  03
 0>  e

™  c
 ftJD'^
 c  c
tS  4>
                         ฃ2
 o
u

-o  .fc.
 C  > :
 03  O  '
 ^  CJ  •
•S  ^ flj-
 e?  ^^^ ^**
*S  P* ซ***

111

 ^1 S
 1   ซ s
                         ฃZ
                                  =  ฎ.2
                                  s  s ^
                                  ซJ  -
-------
ft
ft
  —t    ft
  ^      \ซป
  ft      ^.H
•
Sf
^^
9
^.
1
sr
3

o
-x*
n'
ฃ
B—
ฃL

•ฃ



• '.;
5T

C
>
I
en
3
S
ft
35
ซ
wtf
ft
S*
s*
•••
M
CA
z
*
b
ซ5
- ^
K
1

ft
•? .
ft
ttr-
5*
ซ<
f
•j"
CT!
ft;
S5
• f^-
-
*-<
tf
I
g
n
^
H
S

^/
CA
n
o .
f+- •
n
w*
0-'
^*-
ง•ง•
5
OR
^*
0
•C/i
22
o-
X
ST
-•s ,
?"
5
x"
^^.
•^
s. a • ซ





s.
ft



' &2

S
p^ป
5





                 71

-------
 =
 O'

U
&
D
       ^'— ^^

       w "
-Jv C

ซ 2
&- c
O V2
^
|i
c 5ซ
c: |

ฃ E
— c
       5 ฃ
         •4^

         O
* JS.
^ ^ซ C


ซ *ฃ'*
C  ~ ปT

SSI
u -5 .-

ง c 15
^^ * rf< \ ^-- ^
     c
ซ.    ^
c — w
C5 •  ซ U

gปฐ

            a cs
            56 ง
       Cft
         -.  O
       "j ^ •••
       •*^ C5 <*^

       o e  ซ

       U
            C CQ M
            ซ•ป ** \OA
            & s <&
            CJ
           0)
                             2

e E

|s

tl ฃ
:= c.
J5 U
^^
  •o
   e
o> c:
           dJ  2
           .S-8
           o  i

           ฃง
                        72

-------
ri'ss.S'SJ
          73

-------


/-v
^d
S'
o
U
^•^
en
ELEMENT



s
CA
O
o
^^
.e
1
u
c
mention
Effluent Guidelines
Point Source Controls,
Stormwater
!TT * •
CTJ ^^rii ^^&
2 
-------
on cr
•  •  •
•c > >

ง11
*-<-ซ:<<
<^ •••• •••
3T = s
4g  as w
e  ป ^
w  Cr A-
as  n &

*>1
ซ  "^ ซ

B  ?ฃ
>'ป
*  S
S" as .
3 era

•?  =•
2 TO'

I 53

Hซ
ft  d,
CM ซ3T
^^ ij*}
S' ft
**  ^^
OS  —
                   p
          --H
              •9 S
                        3 a
                   a  oS.
                   "  MM T^
-1. s.-
  •    -
       ft
              s f g.. S g
              •ft *S   C ft
              a.  E. ^ " &
5' B ง
•s  ซ =

   5T
                         Q.,
                         5*
                 H-
                 r
                 R3
                 2
                 t=3
                 Z
                 H
         75

-------
0
         wa

         CJ



         C

        C/2


        "c5

cซ
 Cfl



K
                   ^    c
                          u
                          c
                          V*
                        u    s
                        >   "-5
                                                  0)
            0)


            e

         en
         c
         a

        o
                   C/2
                         =  "8
                         'S  tS

*i   5

 ^   -*^


'S    Is
                                                  C


                                                  u
                                 i
                       ffi--

                       'S
                        en
                                           76

-------
77

-------
•' &
i ฃ2]
H
H ^
^ta ^^^^^^^^
^^ ^•k A
vซ/J "^^-
SESSMENT
(Con
c/^
^^
^
1






ฃ.,
^JQ ^J
.S S
5= K
•3 ซ
•5 o
o • fig
S o
s '





o
^
toring Mission Statcn
's
c
ง

1
c
c
&
^•^
Jm
toring Task Force wit
"s
e
5






' e ' •
System Modernizatio
3
8
C . • -;•

78'

-------

•SB
(t>
      BJ
      S

  H i-'s-i
        i. ง
        ft O
        p c  „
        —• 5"  rt

            a  ป••
            I. 5
0
s:

5
x
s
    fa-  ง
•• n  ^

ฐ I SI  t
?• S
      79

-------


i
1
^"4
^T
O
&•}
^^^^^^* !
^^f^*

Gm
H CT
K5
o ^
:ESEAR
ซ

s
C3
^_
OJD
C
• t.


"S

tS
c
c
C
s
c:
"S Research
w
ฃ













u
22- ซ
S =
^^3 ••
t_ ซ
i • w
eB .2
5 ซ
e ฑi
= i
•fi fi
^^j ^^
^J u








en
C
.ฃ
en
cu
•|
C
lie Workshops
•s
ฃ






C/2

C
eU
ex)
^
"S
•o
CV
•Ss
o
u
CQ
C/2
^
'




•••1
C
&.
en
C
•c
s
^k
c.
c
en
.^en
- ' •*.
ps— Bioassays
••i^
u
C
U
!


s
.2
^^ '
•s
i
1 • '

.2 i
ฃ •
en"
cz
en
en
'ซ
O.
S ' ^
and Wildlife-
^
.ซ
•

-------
    n c  a
    e CA  O
    ง ft  s
ft
   * ft   o

   3 ฃ   S'
   s- —.   3
    S  C
    ฃ0
ac
   ง S  ง

s     c  3
rt •••   1   <*
l*j     ฃJ   ^B^.
=5:   • •ง   ft
55     f*  O
"Z    09  3
 S
 •ซ•*•

 5"
 ป • 88
 = S

 5" •?
 ore 23

 sf
 s ^.

 S 2-

 B-a-
 S G
 S V5
' & ft

 fT ""^

 ฃ ซ?
j ^ "S

•^s  -
ซ" 2 • S*


9s f
ป SB  S
3 "BT "P
g p  J3

ft 7-^-
•   O • 3
   H ^^

   I i
   ft •
                  5A      O


                  5!  ""  ?
                  Q. •    ft
 i
 •ft
 5
       ft
       CA
      i
      •ft'
 2     E?
 *^    P
 _ro     5
 era     ft
    CA


    3



    Q

    ft



    ft*

    E.
    3T
                         ft"
oaccum
                          ft
                                    z
                                    H

                                    O
                                    z
                                    o
     .  81

-------
O
tf
H
PREVE
                 en
                •—  W5
                "ซ  **
                -
            —   .S
             ฃ
                    M
             C
                 c
                 ซ
                 S
e:



fl
ปS
e *E
ซ u
.!ฃ
"s •ซ
x K
^ C3

•=?
                              en

                              ex

                              ซs
                              CA

                              O
                              CJD"

                              EC
                              u
tants
                                  C
                                  C.
                                  u
                              en
E   ฎ
                    wa
             o
            CD
                 e  a)
                 U Z
Learn How t

Bioconcentra
                          "ซ'**—•
                              o   •—
                              CL   Q
                              •C  S

-------
>n
c o .
•a o '
sr •!
o &.
2- s

S2
• -ซ_ ~
& =T
fa
85 ป9
o'S
•"S W5
9w ^^^
'0 <
S ฉ
o =
•' 2 Jf
*• >



5" '
.
E.

^


n
loordinatewit
sr
Z
sป
S"
•ป
ijl Resource IVust
ซ

o"
CO
?r
ft5
cr.
J5-*
S
5"
s
•
55
ccovcr Costs 1
^
•n-
1
ft*
5
&.
K
^•^
^
CA








n
ompel Respor
•V
ft"
ft*
D
s
C
35
5?
Cff






e
Sฃ
n
o
P3
^S^j
^^f
n
r
o
SD
C.
H
C/5
^j
>
&




S'
ปfe
brcement
i
W
CO
rt
d Remediati
o
s







w
P3
l
'ซ•
j^^
.H
S
z
CO
H
r*
H^
r^^
H .
Nrt

^
^

1
                      83

-------
    &.


    "8
    
-------
       e
       CA
N
6?
    ft
    93
  ••ฃ-   ST.
       <
    ป

    i
&
H

ftfS.  I   5'
& i*  ^    s
—•ซ   ^.
                         c?
< o
ft 3'
CA ซ•
<"^ gj

era*. 5
& =.
C. 3
e ป
s r.
•=3

ft
3

ft



W*
        x
             o   O
             5..Z
                         C
  85

-------



ED MATERIAL
DREDG



!x
rtENT STRATEG
[ANAGEP
^
•
-c
DJD
•a
i
ional Guidance on Testing Dre<
•*-*
u
ฃ
Q.
"w
4)
Ct
s.
•*•ป
03
ฃ
V
.s
*u
C5
2
ฃ
C
C5
™
a
^O
."E
CD
u
C
ซฃ
"u
0)


eฐ
to
vised National Guidance on Te
V
S
I
•"ft.



u
1
S
CJ
O
2
^
L.
"3
C
redged Material
C

2
u
e
cut on the Environmental Fact
&
a.
"S
fi



CT!
B
o"
C
^ft.
CA
C
tx
'S
15
onsider When E
U

s
CA
RCRA Disposal Requirements
E
C5
u
ft.
.-
*>
•



V5
^
[a.
"w
^c
ฃ ,
s
^o
]S .
•5 -
rti
gencywide Remi

86

-------
Standardized Sediment Testing:
Needs and Requirements of Key Agency Programs
Thomas Armitage, U.S. EPA Office of Science and Technology


I.     Office of Wetlands, Oceans, and Watersheds

       A.     Oceans arid Coastal Protection Division

              1.    Regulatory Responsibilities:

                    a     Determine  the acceptability of materials, including  dredged
                          materials,  for ocean  dumping under  the  Ocean  Dumping
                          Regulations.

       B.     Statutory Authority

              1.    Marine Protection, Research, and Sanctuaries Act (MPRS A);

              2.    Ocean Dumping Regulations (40 CFR 227 and 228)          ,

       C.     Major Needs and Requirements for Standard Sediment Testing Methods:

              1.    Primary  interest lies in testing dredged material to determine possible
                    effects of disposal. .                                         .

              2    Methods  for  the following  tests should be  standardized:  chemical
                     analytical testing, toxicity bioassays, and bioaccumulation.

              3     A sufficient number of test organism taxa must be standardized in order
                   .  to provide an adequate range of test species and be applicable to a wide,
                     range of sites and contaminants.
                           it is recommended that standardized organisms to be used in acute
                           toxicity tests be from the taxonomic order Amphipoda, and include
                           at lea;t th~ ปrฃ*™ซ™ Ampelisca abdita and RhPT>oxvnms abromus.
      a
      b     Standard organisms for bioaccumulation tests should include a
       '     number  of species from the  following  taxonomic  groups:
            polychaetes (particularly Neanthes sp, Neries sp, and Nepjhys sp),
            moUuscs (particularly Macoma sp), and crustaceans.  Because of
            problems with availability and relative sensitivities, we nee
-------
       5.     Standard methods for use in marine sediment chemistry analysis have been
             developed for tiie Clean Water Act 301(h) program (monitoring ocean
             outfalls of sewage treatment plants), and are the methods specified for use
             in testing sediment for ocean disposal. The Oceans and Coastal Protection
             Division suggests that these methods be adopted for marine sediments
             unless other methods can be standardized and used routinely.

D.     Existing Program Guidance and Tests Used to Assess Sediment Contamination:

       1.     The testing manual, "Evaluation of Dredged Material Proposed for Ocean
             Disposal" outlines procedures for water column and benthic effects tests.

             a.     The testing manual lists suggested and recommended species to be
                    used in specified tests. The decision concerning which species are
                    required for tests currently rests with the EPA Regions and Corps
                    of  Engineers Districts,  which must  consider  local factors in
                    determining species appropriateness.   This is done through the
                    development of local testing manuals.

             b.     Guidance for performing  tests  includes discussion of:  species
                    selection, apparatus, experimental conditions, sample preparation,
                    test design, QA/QC, and data presentation and interpretation.

             c.     Water column tests

                    i.      Elutriate  chemical concentrations  are  ' used  to  assess
                           compliance with water quality  criteria; the water quality
                           criteria   are  considered  the   "Limiting  Permissible
                           Concentration" not to be exceeded after consideration of
                           initial mixing.                      .           .

                    ii.     Elutriate bioassays are static 96 hour LC50  studies using
                           three  concentrations  of  elutriate;  1%  of the LC50
                           concentration is  considered the "Limiting  Concentration"
                           not to be exceeded after consideration of initial mixing.

                    iii.    The  regulations  require  at   least  one  species  from
                           phytoplankton or zooplankton,  crustacean or mollusc, and
                           fish.  A range of test species are suggested, and a subset is
                           recommended, in  the testing  manual for water column
                           bioassays.  However, decisions concerning the choice of
                           test organisms currently rest with local decision makers.

-------
           d.     Whole Sediment Bioassays
                 i      whole sediment bioassays, both acute 10 day bioassays and
                        10 or 28 day bioaccumulation, are described in the testing
                        manual.      . '  .

                 u     For  both acute  10 day  bioassays and  10  or  28 day
                   ''   bioaccumulation, the dredged material results are compared
                        to reference site sediment results.  This is done to ensure
                        that  no unreasonable degradation  will occur  because of
                        disposal.                                    ...-•'.
iii
  '
                        The regulations require that test species comprise filter-
                        feeding, deposit-feeding,  and burrowing  organisms.  A
                        range of  test species  are suggested, and  a subset is
                        recommended, in the testing manual, for whole sediment
                        bioassays. However, the decision concerning choice of test
                        organisms currently rests with local decision makers.

               '••••••'    For acute toxicity bioassays, infaunal amphipods,
                                burrowing polychaetes, molluscs,  and crustaceans
      ,                  .        are suggested.
                         .      For bioaccumulation tests,  polychaetes, molluscs,
                                and crustaceans are suggested.

      2     301(h) monitoring program guidance documents include  a number of
            volumes that may be relevant to contaminated sediment testing:

            a.     301(h)   toxic   effects   of sewage  discharge,   on  coral 'reef
        i           communities.                            ,
            b     Summary  of U.S. EPA approved  methods  to  demonstrate
              "     compliance with applicable water quality  standards.

            c.     QA/QC for 301(h) monitoring program. Guidance on field and lab
                   methods.                    -"
             d      Bioaccumulation series (5 volumes).  Addresses target species,
                    detection limits, analytical methods, and  sample replication.
Wetlands Division

A.     Regulatory Responsibilities:

       1      Develop guidelines to evaluate proposed discharges of dredged or fill
              material into waters of the United States.

                                    89

-------
B.     Statutory arid Regulatory Authority:

       1.     Clean Water Act, Section 404

       2.     Section 404(b) (1) guidelines (40 CFR 230)

C.     Major Needs and Requirements for Standard Sediment Testing Methods:

       1.     Interest lies in testing dredged material to determine possible effects of
             discharge.

       2.     The program would like a tiered testing approach to evaluation of dredged
             material proposed for discharge.  Needs and requirements are similar to
             the ocean disposal program.  However, freshwater and estuarine species
             should also be selected for test development.
                         i                                  - '                 !
D.     Existing Program Guidance and Tests Used to Assess Sediment Contamination:

       1.     Currently there is no broadly applied testing manual for dredged material
             evaluation. Evaluations are handled on a case-by-case basis using 404 (b)
             (1) Guidelines.  However, EPA and the Corps of Engineers  are in the
             process of developing the Inland Testing Manual to evaluate proposed
             discharges of dredged material  into waters inside  the baseline of the
             Territorial Sea. The manual will be completed in 1993.  It incorporates
             a tiered testing strategy.

       2.     The 404  (b) (1)  Guidelines provide a general framework under which
             testing is  to be performed. The general framework  includes: evaluation
             of existing  information, chemical  and  biological  testing  (chemical,
             characterization of material, elutriate testing, and benthic effects testing),
             and evaluation of physical effects of disposal.

       3.     Regional  guidance for testing dredged material has  been developed by:
             Regions 1, 5, 9,  and 10.

             a.     Region 1 protocols have been developed for: selection of sampling
                    sites;  physical and bulk  chemical analysis of sediments;  tiered
                    evaluation testing  for liquid phase assay, suspended paniculate
                    assay,  whole sediment  assay,  and  bioaccumulation  analysis;
                   - elutriate testing procedures; and QA/QC measures.

             b.     Region 9 has developed guidance similar to Region 1 guidance.
                                    90

-------
             c     Region 5 has produced guidance for sampling and testing efforts
                   related to navigational dredging, and also uses the IJC Guidelines
                   and Register for Evaluation of Great Lakes Dredging Projects.

             d.     Region  10  has developed the Puget  Sound  Dredged  Disposal
                   Analysis, which includes a tiered battery of tests.

             e     Other Regions apply variations  of the ocean disposal  testing
                   manual to dredged material assessment programs in waters of the
                   United States.

      4     It should be noted that standardized testing in the ocean dumping, 404,
        'and other programs must address the interfering effects of sediment grain
             size, ammonia, and  hydrogen sulfide.  These  are important issues for
             standardization.


Office of Pollution Prevention and Toxic Substances

A.     Regulatory Responsibilities:

       1      Assessment of risks  resulting from possible releases  of existing and new
              chemicals that are manufactured, distributed, or disposed.

       2.     Decisions to regulate the use of new and existing chemicals.

B.     Statutory Authority:              .

        1.     Toxic Substances Control Act, Section 4, 5,  6, 8

 C.     Major Needs and Requirements for Standard Sediment Testing Methods:

        1     OPPT  is interested in fate, transport and effects of potential sediment
              contaminants. Spiked sediment testing is the kind of standardized test that
              would  be of the greatest usd in these evaluations.

        2     The PCB program is also interested in developing toxicity assays for use
         '     'with sediments taken from sites contaminated with  PCBs.  If a disposal
              method such as bioremediation is used to remove PCBs, toxicity tests are
              necessary to ensure that PCBs have been destroyed, and that intermediates
              more toxic than the original PCBs have not been formed.
                                     91

-------
       3.     Since spiked sediment tests would be of greatest use to the program,
             standardization of the test protocol elements described below -would be
             most useful.

       4.     Sediment testing for one species is described in the program guidelines.
             Standardization  of methods  for additional species would offer a greater
             range of testing options.

D.     Existing Program Guidance and Tests Used to Assess Sediment Contamination:

       1.     TSCA Guidelines used to develop data on the toxicity and bioavailability
             of chemical substances and mixtures.

       2.     Section 795.135 of the Guidelines  describes the chironomid Sediment
             Toxicity Test,

       3.     Chironomid sediment test guideline describes three tests:

             a.     14-day chironpmid aqueous exposure test with minimal sediments,
                    foods, and test substance added to the water.

             b.     14-day exposure with test substance added to the sediment.

             c.     14-day interstitial exposure with  the test substance addecl to the
                    water.

       4.     The guidelines include the following key protocol elements:

             a.     Conducting range finding tests;

             b.     Conducting definitive tests (number of test organisms, specification
                    for controls, test duration, endpoints, water quality measurements);

             c.     Measurement of test substance;

             d.     Test conditions and selection of organisms

                    i.     Acquisition,   feeding,  loading,  care   and  handling,
                           accEmation, facilities;

             e.     Test substance delivery system;

             f.     Dilution water;
                                    92

-------
                   g.     Cleaning test system;
                   h,     Sediments used for test, determination of contaminant partitioning and
     .                     bioconcentration;

                   i.     Additional test parameters and measurements;

               '    j.     Reporting test results.


IV.    Office of Pesticide Programs

       A.    Regulatory Responsibilities:                         ,
             1     Review me uses of new and existing chemicals to be registered as pesticides
                    in order to determine effects on nontarget organisms.

       '     ' 2     Make decisions to: 1) label pesticides in order to control or restrict their use;
                                    ratioofne^^
       •             or
                ban the use of existing pesticides.
B.     Statutory Authority:                                          .   ,
       1.     Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA)

C.     Major Needs and Requirements for Standard Sediment Testing Methods:

       1     The tiesticide program is primarily interested in spiked sediment toxicity tests
             anlS?^
             S^S of Chemicals would affect exposed species.   Standardization of
              spiking methods would therefore be very useful.

       9      The program requires  standardized tests to discriminate among exposure
              SLaysTorS to determine  the bioavailability of contaminants resulting
              from different levels and methods of pesticide application.

        3      OPP is also interested in  evaluating contaminated sediment in the field in
              order to trace runoff of pesticides applied in agricultural practices. .

        4.      Currently,  no standardized sediment toxicity tests have been developed by the
               program for evaluation of pesticides.                         '.-"'••
        5      The program would like to see standardization of marine and fr^terte^
               for boA the acute and chronic studies.  Representative species of appropriate
               Snsiti^ty from a range of sites would have to be available for testing. Acute
               SSSS testing currentiy requires an invertebrate, -da warm water ^d



               probably be similar to those mentioned above.
                                       93

-------
              6.     Mesocosm tests have also been developed by OPP for dose-response
                    studies of pesticides.

              7.     A major concern of the program is that standardized sediment studies
                    would have to be legally defensible if Agency decisions were challenged
                    by the agricultural or chemical industries.

              8.     A major issue for the program in sediment testing is the kind of sediment
                    to use in spiked testing, how the sediment should be handled, and how the
                    contaminant should be introduced to the test system.

              9.     The pesticide testing  guidelines and  standard evaluation procedures
                    developed by OPP provide the level  of detail that would be required in
                    standardization.  This kind of guidance is generally of the same level of
                    detail as an ASTM standard method.

              10.    The level of detail in a standardized test that is likely to be most useful to
                    the pesticide program would be that of the ASTM standard practice.  A
                    standard guide may not offer enough structure, and a standard method
                    may not be flexible enough.

       D.     Existing Program Guidance and Tests Used to Assess Sediment Contamination:

              1.     The pesticide program currently has no existing guidance for sediment
                    testing.  No standard tests or species have been developed. The program
                    has authority, to ask for such a study under special test requirements, but
                    generally has not required sediment bioassays.  If they are required, the
                    pesticide registrant is asked to submit  a protocol for evaluation by the
                    program.

              2.     The Office of Pesticide Programs Standard Evaluation Procedure for
                    Ecological  Risk  Assessment describes how the results of contaminated
                    sediment studies would be used to complete a risk assessment.
V.     Office of Emergency and Remedial Response (Superfund)

       A.    Regulatory Responsibilities:

             1.     Cleanup of hazardous waste sites to protect human health, welfare, and
                    the environment.

             2.     Sediment assessment  methods are  identified and  applied both for site
                    assessments  and remedy selection.


                                          94

-------
       3      Once contaminants are identified, existing state and federal standards are
              evaluated for applicability to the site.  When standards are not available,
              other evaluation methods are used to determine if the environment is
              endangered and to select cleanup goals.

 B.    Statutory Authority:

       1      Comprehensive Environmental Response, Compensation, and Liability Act
         '     (CERCLA),  as  amended  by  the  Superfund   Amendments   and
              Reauthorizatiori Act (SARA).

       2     CERCLA mandates that sampling be conducted to characterize the release
              of hazardous substances from a site  and to determine if these releases
              present a threat to human health, welfare, and the environment.

 C.    Major Needs and Requirements for Standard Sediment Testing Methods:

        1     The program is looking for a battery  of test methods that have been peer
              reviewed  and validated.  These would be used with standard chemical
              analytical methods.  A list  of standardized biological methods that could
              be included in the Superfund Contract Laboratory Program would be very
              useful.
                                           i                       •
        2     The Superfund Program handles a wide variety of sites, requiring a wide
               spectrum of testing options to meet  site specific goals.  More emphasis
;               and resources are being placed on ecological assessment, and the program
               will be doing more quantitative assessment.  Any standard methods that
               can be of use in these assessments will be important to the program.

        3.     The following standardized tests would be useful for the program:

               a. "   Chemical Testing                      -        .

      '            ,    i.     Sediment  analysis   to   determine  bioavailability   of
                            contaminants.

                ,      ii:    Total Organic Carbon (TOC).

                      iii.     Residue Analysis

               b.     Biological Testing

                      i.     Bioaccumulation Tests

                      ii.     Solid Phase Toxicity  Tests
                                      95

-------
      D.     Existing Program Guidance and Tests Used to Assess Sediment Contamination:

             1.     Superfund RI/FS  Guidance and the Environmental Evaluation Manual
                   provide general guidance on methodologies used to determine the nature
                   and extent of sediment contamination at sites. Ecological Assessment of
                   Hazardous Waste  Sites. EPA/600/3-89/013.

             2.     The Data Quality Objectives Guidance and Compendium outline a detailed
                   description of the  investigation process.  A Quality Action Plan (QAP) is
                   developed on each site, and may be carried out by the EPA established
                   contract laboratory program (CLP) or a non CLP laboratory that meets the
                   data quality objectives of the investigation.

             3.     Currently, the Superfund Program draws upon several sources for testing
                   methods.  These include, but are not limited to ASTM, OECD, the U.S.
                   Army  Corps of  Engineers,  EPA Methods for Measuring Acute and
                   Chronic Toxicity  of Effluent Waters, and the Canadian ministry of the
                   Environment. A list of those available for sediment tests is attached.  The
                   ASTM sediment methods are standard guides.

             4.     Region 4 Standard Operating Procedures for Toxicity Testing/Hazardous
                   Waste Assessments.   April  15, 1990.  Written by Todd Harris, Jay
                   Glover, Jim Maudsley, with foreword by Bill Peltier.  U.S.  EPA Region
                   4 Environmental Services Division and NSI Technology Services Corp.  '
VI.    Office of Solid Waste

       A.    Regulatory Responsibilities:

             1.     Under  the  Resource  Conservation  and  Recovery  Act  (RCRA),
                    contaminated sediments exhibiting RCRA toxicity must be managed in a
                    RCRA permitted or interim status facility.

             2.     Authority to enforce cleanup of contaminated sediments is used under two
                  .  conditions: 1) if the sediment is dredged and exhibits a hazardous waste
                    characteristic under RCRA, or if the sediment is mixed with a RCRA
                    listed hazardous waste; and 2) if the sediment contamination can be shown
                    to have resulted from a release form a specified solid waste management
                    unit at a RCRA permitted or interim  status hazardous waste facility.

             3.     Contaminated sediments  may be toxic under the toxicity characteristic
                    leaching procedure test, a test that compares the concentrations of various
                    chemicals in the  leachate .from  the dredged  materials  with levels
                    established to protect the environment.
                                          96

-------
1.
     B.     Statutory Authority:                        ,

            1.     Resource Conservation and Recovery Act (RCRA), Sections 3004 (u) and
                  (v), and  Section 7003.

     C.     Major Needs and Requirements for Standard Sediment Testing Methods:

                  If new sediment quality assessment methods receive adequate scientific
                  review, OSW would incorporate them in making RGRA permit decisions,
                  and in evaluation of remedial site restoration plans.

            2     The program would be interested in tests that can link toxicity to a source
             'of contamination.  Toxicity identification evaluation would be useful.

      D.    Existing Program Guidance and Tests Used to Assess Sediment Contamination:

             1      The  program relies on the Superfund guidance described  above   No
              '     standard program  approaches  to   evaluating   or   testing  sediment
                   contamination  have been developed.  Testing decisions are made on  a
                   case-by-case basis. The program would be interested in a range of testing
                   options  that could provide the appropriate choice for a particular site.


Vri.  Office of Wastewater Enforcement and Compliance

      A.  '  Regulatory  Responsibilities:

             1      NPDES permits issued  under the Clean Water  Act can be written  to
              '     protect  against sediment contamination.  Sediment testing and monitoring
                    can  be  required as a condition of a discharge permit.

             2      At present, water quality  based effluent limits protect against sediment
                    contamination only to the extent that such contamination would cause
                    violations  of water quality criteria.

       B. ..  Statutory Authority:

              1.     Clean Water Act, Sections 101(4) (3), 301 (b) (1) (C), 402, and 304 (1).

       C.     Major Needs  and Requirements for Standard Sediment Testing Methods:

              1.     Acute  and chronic sediment bioassays and bioaccumulation studies could
                     be  used in the NPDES  permitting process.
                              97

-------
D.     Existing Program Guidance and Tests Used to Assess Sediment Contamination:

       1.     Sediment quality  criteria are being developed to be used in NPDES
             permitting. .        -           '    '

       2.     The guidance document, "Assessment and Control of Bioconcentratable
             Contaminants in Surface Waters,"  March 1991, describes methods for
             sampling and measuring bioconcentratable chemicals in sediment and biota
             at point source discharge sites.
                                   98

-------
      PROGRAM OFFICES
      RESPONSIBLE FOR
SEDIMENT TOXICITY TESTING

 . OFFICE OF WETLANDS, OCEANS, AND WATERSHEDS
  - OCEANS AND COASTAL PROTECTION DIVISION
  - WETLANDS DIVISION
                       i
 • OFFICE OF WASTEWATER ENFORCEMENT AND
  COMPLIANCE
 • OFFICE OF POLLUTION PREVENTION AND TOXIC
  SUBSTANCES
 . OFFICE OF PESTICIDE PROGRAMS
 • OFFICE OF EMERGENCY AND REMEDIAL RESPONSE
                               'ซ
 .  OFFICE OF SOLID WASTE
                  99

-------
   OFFICE OF WETLANDS, OCEANS , AND
  WATERSHEDS - OCEANS AND COASTAL
         PROTECTION DIVISION

STATUTORY AUTHORITY AND REGULATORY
            RESPONSIBILITY
    DETERMINE ACCEPTABILITY OF MATERIALS
    (INCLUDING DREDGED MATERIALS) FOR
    OCEAN DUMPING

    MARINE PROTECTION RESEARCH AND
    SANCTUARIES ACT (MPRSA)

    OCEAN DUMPING REGULATIONS (40 CFR 220-229)
                   100

-------
        OCEANS AND COASTAL
PROTECTION DIVISION REQUIREMENTS
 FOR STANDARD SEDIMENT TESTING
              METHODS

v  CHEMICAL ANALYTICAL TESTING, TOXICITY
  BIOASSAYS, AND BIOACCUMULATION TESTS
v  ORGANISM TAXA MUST BE APPLICABLE
  TO WIDE RANGE OF SITES AND CONTAMINANTS
   RECOMMEND USE OF AMPHIPODS FOR ACUTE
   SEDIMENT TOXICITY TESTS
   - AMPELISCA ABDITA. RHEPQXYNIUS ABRQNIUS
    RECOMMEND SEVERAL TAXA FOR
    BIOACCUMULATION TESTS
    -- POLYCHAETES (NERJES SPECIES)
    - MOLLUSCS (MACOMA SPECIES)
    WHOLE SEDIMENT CHRONIC BIOASSAYS
    NEEDED FOR OCEAN DUMPING PROGRAM
    SEDIMENT CHEMISTRY METHODS DEVELOPED
    FOR CWA 301 (H) PROGRAM
                    101

-------
 OCEANS AND COASTAL PROTECTION
DIVISION EXISTING TESTING GUIDANCE
 TESTING MANUAL, "EVALUATION OF DREDGED
 MATERIAL PROPOSED FOR OCEAN DISPOSAL" -
 "THE GREEN BOOK"
 LISTS SUGGESTED AND RECOMMENDED SPECIES
 FOR TESTS
  PROVIDES GUIDANCE FOR PERFORMING TESTS
  - SPECIES SELECTION
  -- APPARATUS
  -- EXPERIMENTAL CONDITIONS
  -- SAMPLE PREPARATION
  -- TEST DESIGN
  -- QA/QC REQUIREMENTS
  -- DATA PRESENTATION AND INTERPRETATION
                    102

-------
    "GREEN BOOK" TESTS






ELUTRIATE BIOASSAYS
                1     . •

- STATIC 96 HR LC50 STUDIES



 WHOLE SEDIMENT BIOASSAYS



-- 10 DAY BIOASSAYS


- 10 OR 28 DAY BIOACCUMULATION TESTS
    ..'   .•"'   .. '     .        '    • \

DREDGED MATERIAL RESULTS ARE

COMPARED TO REFERENCE SITE SEDIMENT

TEST RESULTS
                103

-------
   OFFICE OF WETLANDS, OCEANS, AND
       WATERSHEDS -- WETLANDS
                DIVISION

STATUTORY AUTHORITY AND REGULATORY
            RESPONSIBILITY
     TESTING DREDGED MATERIAL TO DETERMINE
      EFFECTS OF DISCHARGE

     CLEAN WATER ACT SECTION 404

     SECTION 404(B)(1) GUIDELINES
      (40 CFR 230)
                    104

-------
  WETLANDS DIVISION EXISTING
       PROGRAM GUIDANCE
.  CURRENTLY NO BROADLY APPLIED TESTING

  MANUAL
   EVALUATE PROPOSED DISCHARGES OF
   DREDGED MATERIAL

.  404(BX1) GUIDELINES PROVIDE GENERAL
   TESTING FRAMEWORK
    MATERIAL TESTING
                  105

-------
OFFICE OF POLLUTION PREVENTION
     AND TOXIC SUBSTANCES

   STATUTORY AUTHORITY AND
  REGULATORY RESPONSIBILITY
 ASSESSMENT OF RISKS FROM RELEASE
 OF EXISTING AND NEW CHEMICALS
 THAT ARE MANUFACTURED, DISTRIBUTED,
 OR DISPOSED

 TOXIC SUBSTANCES CONTROL ACT, SECTIONS
 4, 5, 6, AND 8
               106

-------
OFFICE OF POLLUTION PREVENTION. ^


             METHODS
 INTERESTED IN FATE, TRANSPORT, AND EFFECTS
 OF SEDIMENT CONTAMINANTS

 SPIKED SEDIMENT TESTING IS OF GREATEST USE

 PCB PROGRAM INTERESTED IN TOXICITY ASSAYS
  FOR PCBS
                   107 .

-------
OFFICE OF POLLUTION PREVENTION
 AND TOXIC SUBSTANCES EXISTING
        TESTING GUIDANCE
SECTION 795.135 OF TSCA GUIDELINES
 DESCRIBE CHIRONOMID SEDIMENT TOXICITY
TEST

TEST GUIDELINES DESCRIBE THREE TESTS

 - 14 DAY CHIRONOMID AQUEOUS EXPOSURE
  TEST WITH MINIMAL SEDIMENTS AND FOOD
  ADDED TO THE WATER

 -14 DAY CHIRONOMID TEST WITH TEST
   SUBSTANCE ADDED TO SEDIMENT

 -- 14 DAY INTERSTITIAL EXPOSURE WITH
   TEST SUBSTANCE ADDED TO THE WATER

 GUIDELINES INCLUDE A NUMBER OF KEY
 PROTOCOL ELEMENTS
                 108

-------
OFFICE OF PESTICIDE PROGRAMS

  STATUTORY AUTHORITY AND
  REGULATORY RESPONSIBILITY
—^ •      _    "    ' - '

  REVIEW USE OF NEW AND EXISTING
  CHEMICALS REGISTERED AS PESTICIDES
  TO DETERMINE EFFECTS ON
  NONTARGET ORGANISMS

   MAKE DECISIONS TO LABEL PESTICIDES
   TO RESTmCT USE, PROHIBIT REGISTRATION
   OR USES  OR CANCEL/BAN USE OF EXISTING
   PESTICIDES
   FEDERAL INSECTICIDE, FUNGICIDE, AND
   RODENTICIDE ACT (FIFRA)
                 109

-------
OFFICE OF PESTICIDE PROGRAMS
 REQUIREMENTS FOR STANDARD
  SEDIMENT TESTING METHODS
  PRIMARILY BNTERESTED IN SPIKED SEDIMENT
  TOXICITY TESTS TO DEVELOP DOSE/RESPONSE
  RELATIONSHIPS

  REQUIRES TESTS TO DISCRIMINATE AMONG
  EXPOSURE PATHWAYS

  INTERESTED IN EVALUATING CONTAMINATED
  SEDIMENT IN THE FIELD TO TRACE
  PESTICIDE RUNOFF

  PROGRAM NEEDS ACUTE AND CHRONIC MARINE
  AND FRESHWATER TESTS

  REPRESENTATIVE SPECIES FROM A RANGE
  OF SITES ARE NEEDED

   MAJOR ISSUE FOR PROGRAM IS KIND
   OF SEDIMENT TO USE IN SPIKED TESTING
   AND HOW TO INTRODUCE CONTAMINANT
   INTO TEST SYSTEM
                 no

-------
OFFICE OF PESTICIDE PROGRAMS
  EXISTING TESTING GUIDANCE



OPP CURRENTLY HAS NO EXISTING GUIDANCE FOR
SEDIMENT TESTING

PROGRAM HAS AUTHORITY TO ASK FOR SEDIMENT
 STUDY UNDER THE SPECIAL TEST
 REQUIREMENTS

 OPP STANDARD EVALUATION PROCEDURE
 FOR ECOLOGICAL RISK ASSESSMENT  _
 npQPRlRES HOW RESULTS OF SEDIMENT
 STUDIES WOULD BE USED IN RISK ASSESSMENT
                 111

-------
  OFFICE OF EMERGENCY AND
     REMEDIAL RESPONSE

  STATUTORY AUTHORITY AND
 REGULATORY RESPONSIBILITY
CLEANUP OF HAZARDOUS WASTE SITES
TO PROTECT HUMAN HEALTH, WELFARE,
AND THE ENVIRONMENT

SEDIMENT ASSESSMENT METHODS
 APPLIED FOR SITE ASSESSMENT AND
 REMEDY SELECTION

EXISTING STATE AND FEDERAL STANDARDS
 USED TO SET CLEANUP GOALS, IN
 ABSENCE OF STANDARDS, OTHER ASSESSMENT
 METHODS USED

COMPREHENSIVE ENVIRONMENTAL RESPONSE,
 COMPENSATION, AND LIABILITY ACT
 (CERCLA) AS AMENDED BY SUPERFUND
 AMENDMENTS AND REAUTHORIZATION ACT
 (SARA)

CERCLA MANDATES SAMPLING TO
CHARACTERIZE RELEASE OF HAZARDOUS
 SUBSTANCES FROM A SITE
                112

-------
OFFICE OF EMERGENCY AND REMEDIAL
    RESPONSE REQUIREMENTS FOR   ^
STANDARD SEDIMENT TESTING METHODS
   AND VALIDATED
   MORE EMPHASIS IN PROGRAM IS BEING
   PLACED^ ECOLOGICAL ASSESSMENT


                  113

-------
 OFFICE OF EMERGENCY AND REMEDIAL
RESPONSE EXISTING TESTING GUIDANCE
   SUPERFUND Rl/FS GUIDANCE AND ENVIRONMENTAL
   EVALUATION MANUAL

   DATA QUALITY OBJECTIVES GUIDANCE AND
   COMPENDIUM

   OTHER SOURCES OF TESTING METHODS INCLUDE

   - ASTM, ARMY COE METHODS, EPA METHODS
     FOR MEASURING EFFLUENT TOXICITY,
     CANADIAN MINISTRY OF ENVIRONMENT

   EPA REGION 4 STANDARD OPERATING       .^
   PROCEDURES FOR TOXICITY TESTING/ HAZARDOUS
   WASTE ASSESSMENTS
                    114

-------
    OFFICE OF SOLID WASTE


  STATUTORY AUTHORITY AND

 REGULATORY RESPONSIBILITY



RESOURCE CONSERVATION AND
RECOVERY ACT (RCRA) SECTIONS 3004 (U)
AND (V) AND SECTION 7003

PROGRAM DETERMINES WHETHER
CONTAMINATED SEDIMENT SHOULD BE^
MANAGED IN RCRA PERMITTED FACILITY

PROGRAM HAS AUTHORITY TO ENFORCED _
CLEANUP OF SEDIMENT IF IT IS HAZARDOUS
WASTE UNDER RCRA OR IF IT HAS BEEN
RELEASED FROM A RCRA PERMITTED FACILITY
                 115

-------
OFFICE OF SOLID WASTE REQUIREMENTS
   FOR STANDARD SEDIMENT TESTING
               METHODS
    PROGRAM IS INTERESTED IN TESTS LINKING
    TOXICITY TO A SOURCE OF CONTAMINATION

    IF NEW SEDIMENT ASSESSMENT METHODS
    ARE DEVELOPED THEY WOULD BE USED
     IN MAKING RCRA PERMIT DECISIONS, AND
    IN EVALUATING REMEDIAL SITE RESTORATION
    PLANS
                   116

-------
OFFICE OF SOLID WASTE EXISTING
       TESTING GUIDANCE

       _——	—	:	;	

NO STANDARD PROGRAM APPROACHES
TO EVALUATING SEDIMENT

PROGRAM RELIES ON SUPERFUND GUIDANCE

TESTING DECISIONS ARE MADE ON A CASE-BY-
 CASE BASIS
 A RANGE OF TESTS WOULD BE NEEDED TO
 PROVIDE AN APPROPRIATE TEST FOR
 AN INDIVIDUAL SITE
                  117

-------
 OFFICE OF WASTEWATER ENFORCEMENT
            AND COMPLIANCE

STATUTORY AUTHORITY AND REGULATORY
             RESPONSIBILTY
     ISSUES NPDES PERMITS UNDER CLEAN WATER
     ACT

     PERMITS CAN BE WRITTEN TO PROTECT AGAINST
     SEDIMENT CONTAMINATION

     SEDIMENT TESTING AND MONITORING CAN
     BE REQUIRED AS CONDITION OF A DISCHARGE
     PERMIT

     CLEAN WATER ACT SECTIONS 101 (A)(3),
     301(B)(1)(C),AND304(L)
                     118

-------
OFFICE OF WASTEWATER ENFORCEMENT
   AND COMPLIANCE REQUIREMENTS
   FOR STANDARD SEDIMENT TESTING
              METHODS
    ACUTE AND CHRONIC SEDIMENT BIOASSAYS
    AND B10ACCUMULATION STUDIES COULD
    BE USED IN NPDES PERMITTING PROCESS

    SEDIMENT QUALITY CRITERIA NOW
    BEING DEVELOPED TO BE USED IN NPDES
    PERMITTING
                    119

-------
OFFICE OF WASTEWATER ENFORCEMENT
  AND COMPLIANCE EXISTING TESTING
               GUIDANCE
    "ASSESSMENT AND CONTROL OF
    BIOCONCENTRATABLE CONTAMINANTS IN
    SURFACE WATERS"

    DESCRIBES METHODS FOR SAMPLING AND
    MEASURING BIOCONCENTRATABLE
    CHEMICALS IN SEDIMENT AND BIOTA AT
    POINT SOURCE DISCHARGES

    SEDIMENT CRITERIA METHODOLOGY
    (EQUILIBRIUM PARTITIONING METHOD)

    MODELS TO BACK CALCULATE PERMIT LIMITS
    FROM SEDIMENT CRITERIA
                    120

-------
EPA Regional Sediment Needs                                    .
William Peltier, U.S. EPA Region TV, Environmental Services Division - Athens, GA
I.     Present Regional Activities

       A.     Superfund
       B.     MPDES
       C.     Dredge material
       D.     Special investigations
       Standardized Test Methods

       A.     ASTM
       B.     EPA
       C.     COE/EPA
       D.     Modification of existing methods
 HL   Reference Sediment

       A.    Regional periodic reference area
       B.    Synthetic reference sediment
 IV.    Control Sediment

        A.    Synthetic sediment
        B.    Site sediment
        C.    Regional site sediment
  V.    Species Selection

        A.     Standard test species
        B.     Regional test species
        C.     Criteria for alternate test species*
  VL   Reference Toxicant Testing

        A.    Selection of chemicals for reference testing
        B,    Required series of chemicals used in reference testing for Regional or alternate
               test species                                   ,
                                           121

-------
VH.  Test Conditions

    '  A.    Summary of test conditions and test acceptability for each selected test species
    .  QA/AC Program

       A.    Contract laboratory evaluations
       B.    Accreditation of laboratories'
IX.    Sample Collection, Preservation and Holding

       A.     Consistency in collection methods
       B.     Preservation and holding times of sediment
 X.    Bioaccumulation                                      >

       A.    Standardized sampling protocols
       B.    Minimum detection levels with available analytical methods
 XI.   Technical Transfer

       A.     Regional and State program assistance


 XH.  Regional Resources

       A.     Present Regional and State staffing
       B.     Facilities and future initiatives
                                          122

-------
•a- 5-
>2?
Z'cft-m
zS
gg
33 r
O.
O
>
c/>
-H.:. '
P"
I
O
w
T3
O
0)
>
O
H
•
D
S3
o
5
ง
o
m
C/3
^p *







o
  123

-------
05
Q
O
LU
Q
CC
Q
CO
         LU
         O
      CO
      Q


      Sป
      Lฑ Q
      LU O
•••^^H








<
O LU
^ 2 >:
i_ -> en
co ^ O
      LU
         u_
       DC
   O Q LU  CC
      I-
         Lu og
         co O ^  Q
             174.

-------
125

-------
          LLJ
Q
LU
CO
LU
O
LLJ
DC
LU
LJ_
LU
DC
                 CO
                 LU
                 b co
                 CO LU
                 LU b
                 O CO
                    LU
                          CO
                 0 "  2
                       ^ cc
                 ฃE
              CO
la
                    126

-------
127

-------
o
CO
LU
or
LU
LU
DC

O
Q.
                CD
      LLJ
      h-
      h-
   zs
oPl
<
DC

fe CLJ
      O
   LU ^
   cc
CO CO CO
              128

-------
129

-------

























CD
— -^
_
l^^™1
0)
UJ

l~~
^^
~p
^^
o
X
o
h-
•
LLJ
O
21
LLJ
o:
LU
u_
LLJ
o:
H
•
/^•fcXffi
(• ^^^^8^/s
CD x^_JX ฐ"
^^ ,
Z P
^ LLJ
Q LLJ
LU O
' CO 1- Z •
,i, CO LU
^^^^1 III ^^rfW
X X IrJLJ f-T
^: ^ H- LLJ
LLJ LJJ W m
h- CC ^ rr
•37- LJJ ^ .
-^ป PI ULJ LJL
• S -LIJ DC O
^ CC LLJ s.
— 1 ^ LL ?t
O Q LLJ ^
f*\ LL. rr ^-
***** ^^ LI i "•••*
LL. Cl ^\ ^3
\n -^ Tฃ r*i
i <ฃ- rn >ซ/
t? < J5 LJJ
^ H ? DC
^ CO C/5 U-



1


























130

-------
•*
03

O
      oo
      o
TI
rn
    O

    3
    >
    o
    o
    33

    m
m
o

HI
— n
    o
 o
             D
              D
              O
O
Q
 = ง.ป
         131

-------

-------
133

-------

-------
135

-------
f

-------
Tiered Sediment Testing Conceptual Overview
Elizabeth Southland, 17.5. EPA Office of Science and Technology
       ThP office of Water (OW), the Office of Pesticide Programs (OPP), the Office of







 gSSwoufd be developed to explain how a decision could be made at the end of each tier
 on whether a sediment poses a risk that would trigger a regulatory action.
                                          137

-------
138

-------
o
-n
-n
0
m
O
-n
EMERGE
z
0
-<
^
z
^^^_
0
-n
O
m
O
Tl
(fl
o
r-
o
S
^
CO



o
-n
-n
O
m
o
•n
POLLUTK
o
z *
-o
30
m
o
Tl
Tl
O
m
O
Tl
PESTICID
m
*n
w
0
Q
O
Tj
O"
m
Tl
_f
•21
m
30





o

30
m
m
                                  139

-------
Ill
O
O
   yj
   UJ
C/)Q
O
O
OJCO
งE
ox
LUh-
           
-------
141

-------
LU
O
<
a
        CO
    U_Q
        LU
Q.
DC
LU
                LL =

                O

                $U1
                2- {A
                III g
                Q LLJ
                COLU
                0
                m-  ,*
                OLUO
                     CO
                Q.O<
                X<
                UJLJJ
                           LU
                           O
O
O
LL,

O
LU
Q.
                           GC
                           o
                           o
                           DC
                           a.
                                LLJ
                                o
HE
2

LU
LL
o
                                X
                                0
                                LU
           LL
           cn
           CO
                CT)
                z
                o
                q
                E
Q.
s
o
o
LU
o
           LU
           a:
           LU
           LL.
           LU
           a:
                      142

-------
                         H
                         m
                         30
I
C/J
m
CO
TJ
m
o
o
-n
m
o
f/v
STUDIES



•
30
m
"n
m
30
m
o
m
30
m
o
O
•si
2
"O
>
3D
0)

0)

ACUTE TO
BIOASSA^
X
^^^ *^^
^^^ •••
0>
mnj
3D —
>0
^S
mo
00
S

^
WATER QL
QUALITY C
^^M ^"^
HC
rriH
2 <
Oฐ
^0)
EDIMEN'
IPARISO
z-1
O)

CHEMICAL
a
i
o
m
z
rn
33
B^ป
m
a



REVIEW OF
BIOLOGICA
OF NEARB\
7:r-m
••|5&
ง>i
m>o
•wง-0
5x
O m
30^
15
• m-1"
z>
oi
30
H
o
              143

-------
Q
LLI
DC
UJ
HI
o
O
X
o

UJ
H

o
     o

     CO
     DC
      x
      o
      Qlil

      o<
             ^" ^
             UJ

             o

             o
             UJ
             u.
      H UJ    t 111
              O uj
              O ULJ
              S0

              0<
                     o
                     CO
                     UJ
CO
O
o
o
0)
111
s
DC
O
o

UJ
LL.
 DC
 UJ
      CM

      DC
      UJ
              CO

              DC
              UJ
DC
UJ

H
           144

-------
145

-------
146

-------
Summary of ASTM Activities on
                               Freshwater and Marine Sediment Test Methods
     Objectives

     A.    Overview ASTM subcommittee E47.03 on sediment toxicology
     B.    Standardization of sediment testing


     Advantages of Standardization

     A. ••' Useof uniform testing procedures (e.g., dilution water, duration)
     B.    Increase data accuracy and precision
     C.    Facilitate test replication
     D.    increase comparative value of results
     E.    Greater regulatory and legal impact


     Disadvantages of Standardization

      A     Sediment testing in infancy relative to aquatic testing


      ?:   : SSSSSiSSSi - -*ป — * ,,*ป


IV.   American Society for Testing and Materials

      A     Goal- "Develop standards on characteristics and performance"of products,
            svstems   and s^s^promotion of related  knowledge    -voluntary
            conSsus/  Publication:  Annual Book of ASTM Standards, Volume   ,

            1L04


V.   ASTM Definitions

      A.   Standard: Document development within principals of the society =

            consensus                       '',."•*  ^nn
      B    Guide:  Series of options, no recommended course of action
.   ,    C.   Test Method: Definitive procedure for measuring characteristics
     f                                            .        '   .
                                       147

-------
VI.   Balloting a Document by Voting Members

      A.    Consensus at 4 levels
             1.     Task group (1 to 10 individuals                   ;
             2.     Subcommittee (100 individuals + 500)
             3.     Main committee (250 individuals)
             4.     Society (33,000 individuals)
      B.    Re-ballot every 3 years (at minimum)

VH.  ASTM E47.03 SedJment Toxicology Subcommittee

      A.    Inception:  May 1987
      B.    Goal:  "Develop guides for assessing the bioavailability of contaminants
             associated with sediments ... evaluate hazard of contaminated sediment,
             soil, sludge, drilling fluids, and similar materials."
      C.    Meeting schedule:   Spring - during annual ASTM symposium
                                 Fall - weekend before SETAC
      Approved ASTM Standards
vm.
       A.    E 1367-92:
             Amphipods
       B.    E 1383-92:  Standard Guide .
       C.    E 1391-90:   Standard Guide
             Manipulation
                         Standard Guide ... 10-d Toxicity with Marine & Estuarine

                                          Toxicity with Freshwater Invertebrates
                                            Collection. Storage.  Characterization.
K.    Documents in Balloting Process (task Groups)

       A.     Designing Biological Tests (Dwyer; Main/Subcommittee)
       B.     Toxicity Tests with Polychaetes (Reish; Main/Subcommittee)
       C.     Toxicity Tests with Mayflies (Bedard; Main/Subcommittee)
       D.     Terminology (Ingersoll; Main/Subcommittee)
       E.     Bioaccumulation by Benthic Invertebrates (Lee; Subcommittee)
       F.     Bioaccumulation by Fish (Mac; Subcommittee)

X.    Documents Proposed

       A.     Toxicity Tests with Oysters (Dinnel)
       B.     Toxicity Tests with Echinoderms (Dinnel)
       C.     Toxicity Tests with Earthworms (Callahan)
       D.     Toxicity Tests with Microtox (Evereklian)
       E.     Toxicity Tests with Tubifex tubifex (Day)
       F.     Sediment Resusoension  (Burton)
       G.     Statistical Guidance (Schlekat)
       H.     Toxicitv Identification and Evaluation (??)
                                          148

-------
XI.   E 1367:  Marine and Estuarine Amphipods

      A.    Scope, significance, and use
      B.    Interference
      C     Hazards
      D.    Test water, test and control sediments, test organisms
      E.    Experimental design, procedure, analyses
      F.    Acceptability and interpretation                      .
       G.    Species-specific 'annexes

 XII.   E 1367:  Marine and Estuarine Amphipods

       A.    Species-specific annexes    •
             Annex!.     Rhepoxvnius abronius
             Annex 2.     Eohaustorius spp.
             Annex 3.     Ampelisca abdjta
             Annex 4.     flrandidierella japonica
             Annex 5.     T^ptocheirus plumulosus
             Annex 6.     Hyalella azteca (proposed)      •

 XII    E1367: Marine and Estuarine Amphipods


        A'    ^    To-d whole sediment, static, field contaminated/spiked sediment
              2.    Endpoints ^survival, reburial
        B.    Status:  approved standard
        G.    Future plans:                                              ;
               1.    Chronic endpoints (growth, reproduction)
              2.    "Rhepoxynius abronius test method?

  Xin.  E 1383:  Freshwater Invertebrates

        A.     Species-specific annexes
               Annex!.    ; HyjMla azteca                      ,
               Annex 2.     Chironomus tentans                   :
               Annex 3.     Chironomus riparius        .                    .
               Annex 4.     Daphina sp. and Ceriodaphnia sp.
               Annex 5.     Hexagenia sp. (Main/Subcommittee)         •   _
               Annex 6.     Diporeia SP. (proposed;  formerly Pontoporeia hoyj)
               Annex 7.     T.umbriculus sp. (proposed)
               Annex  8.     Oligochaeta (proposed, Jubifex tubifex)
               Annex  9.     MoUusks (proposed)
                                           149

-------
XV,   E 1383: Freshwater Invertebrates

       A.    Procedures:
             1.     Partial life cycle, static/flow-through, field contaminated/spiked
                    sediment
             2.     Endpoints = survival, growth, reproduction
       B.    Status:  approved standard
       C.    Future plans:
             1.     Additional species-specific annexes
             2.     Hyalella azteca test method?

XVI.  E1391:  Collection, Storage, Characterization, Manipulation

       A.    Procedures:
             1.     Sediment collection, transport, storage, characterization, spiking,
                    and dilution
             2.     Recommendations and limitations
       B.    Status:  approved standard
       C.    Future plans:  test methods?

XVH. Designing Biological Tests with Sediment

       A.    Procedures:
              1.     Test type (e.g., whole sediment, pore water,.elutriate)
             2.     Sample collection, handling, and manipulation
             3.     Test organisms and endpoints
             4.     Experimental design
             5.     Statistics,  data interpretation,  QA/QC
       B.     Status:  Main/subcommittee ballot

XVm Sediment Toxicity Tests with Polychaetes

       A.     Procedures:                             •              ,
              1.     4 to 20-d test, juvenile/adult, field contaminated/spiked sediment
              2.     Endpoints = survival, growth
       B.     Status:  Main/subcommittee ballot

XIX.  Bioaccumulation Bv Benthic Invertebrates and Bioaccumulation By Fish

       A.     Procedures:                            .
              1.      10 28-d, static/flow-through,  field contaminated/spiked sediment
              2.     Bioaccumulation potential vs. steady  state                    ,
       B.     Status: Subcommittee ballots
                                          150

-------
XX.  Research Needs for Standard Development

      A;    Multi-'species comparisons
      B.    Inter-laboratory comparisons
      C. -   Abiotic factors
      D.    Life history and chronic indicators of toxicity
      E.    Spiking methods and positive controls
      F.    Dilution studies and mixtures
       G.    Laboratory to injitu comparisons
                                           151

-------

-------
               O

               0
                        O
                        ฃ
Q.

N

^*

O
3

O
               (D


               >

               23
                  m
       O       (/>
       ^*      *"
 0
 a

 ro
 3

 5       -
 to       m
 *•*       i^
               IT
               O
               O


               3

               S
               (D
       (Q
                O
                CO
                cn
                o
                a


                0
                 X.

                 O
                 O

                 O
                 CD
153

-------
ADVANTAGES OF STANDARDIZATION




•     Use of uniform testing procedures (e.g.; dilution water, duration)




•     Increase data accuracy and precision




•     Facilitate test replication




•     Increase comparative value of results




•     Greater regulatory and legal impact
                                   154

-------
DISADVANTAGES OF STANDARDIZATION





•    Sediment testing in infancy relative to aquatic testing




•    Inhibit creative approaches



•    inadequate characterization of effects because of optimized conditions
                                    155

-------
AMERICAN SOCIETY FOR TESTING AND MATERIALS
     GOAL:
"Develop standards on characteristics and performance of
                     products, systems, and services; promotion of related
                     knowledge...voluntary consensus."
     PUBLICATION:    Annual Book of ASTM Standards, Volume 11.04
                                  156

-------
ASTM DEFINITIONS





•    STANDARD:
     GUIDE:
     TEST METHOD:
     DOCUMENT DEVELOPED WITHIN PRINCIPALS OF THE





     SOCIETY = CONSENSUS




     SERIES OF OPTIONS, NO RECOMMENDED COURSE OF
                       ACTION
     DEFINITIVE PROCEDURE FOR MEASURING





/     CHARACTERISTICS
                              157

-------
BALLOTING A DOCUMENT BY VOTING MEMBERS





•    CONSENSUS AT 4 LEVELS




     1.    Task Group (1 to 10 individuals)




     2.    Subcommittee (100 individuals+500)




     3.    Main Committee (250 individuals)




     4.    Society (33,000  individuals)




•    RE-BALLOT EVERY 3 YEARS (at minimum)
                                   158

-------
ASTM E47.03 SEDIMENT TOXICOLOGY SUBCOMMITTEE





•     INCEPTION: May 1987




•     GOAL:     "Develop guides for assessing the bioavailability of contaminants




                associated with sediments... evaluate hazard of contaminated




                sediment, soil, sludge, drilling fluids, and similar materials."




•     MEETING SCHEDULE:




                Spring-During Annual ASTM Symposium




                Fall-Weekend before SETAC
                                    159

-------
APPROVED ASTM STANDARDS
                                     ป
•    E 1367-92: Standard Guide ... 10-d Toxicity with Marine & Estuarine Amohipods .

•    E 1383-92: Standard Guide ... Toxicity with Freshwater Invertebrates

•    E 1391-90: Standard Guide ... Collection. Storage. Characterization. Manipulation
                                    160

-------
DOCUMENTS IN BALLOTING PROCESS (Task Groups)




•   - reigning Biological Tests (Dwyer; Main/Subcommittee)




•    Toxicity Tests with Polvchaetes (Reish; Main/Subcommittee)




•    Toxicity Tests with Mayflies (Bedard; Main/Subcommittee)




•    Terminology (Ingersoll; Main/Subcommittee)




•     Bioaccumuiation *y Ronthin invertebrates (Lee; Subcommittee)




•    Bioaccumuiation by Fish (Mac; Subcommittee)
                                161

-------
DOCUMENTS PROPOSED





•    Toxicity Tests with Oysters (Dinnel)




•    Toxicity Tests with Echinoderms (Dinnel)




•    Toxicitv Tests with earthworms (Callahan)




•    Toxicity Tests with Microtox (Evereklian)




•    Toxicity Tests with Tubifex tubifex (Day)




•    Sediment Resuspension (Burton)




•    Statistical Guidance (Schlekat)




•    Toxicitv Identification and Evaluation (??)
                                   162

-------
E 1367: MARINE AND ESTUARINE AMPHIPODS





A.   Scope, Significance, and Use




B.   Interferences




C.   Hazards



p   Test Water, Test and Control Sediments, Test Organisms




.E..' •Experimental-Design, Procedure, Analyses




 F.    Acceptability and Interpretation




 G.   Species-specific Annexes
                                   . 163

-------
E 1367: MARINE AND ESTUARINE AMPHIPODS
     SPECIES-SPECIFIC ANNEXES
     Annex 1.    Rhepoxynius abronius
     Annex 2.    Eohaustorius spp.
     Annex 3.   Amoelisca abdita
     Annex 4.   Grandidierella iaponica
     Annex 5.    Leotocheirus plumulosus
           6.    Hvaiella azte'ca (proposed)
                                    164

-------
E 1367: MARINE AND ESTUARINE AMPHIPODS





•    PROCEDURES:




     -10-d whole sediment, static, field contaminated/spiked sediment




     •Endpoints = survival, reburial




•   STATUS: approved standard




•    FUTURE PLANS:



      -Chronic endpoints (growth, reproduction)





      -Rhepoxvnius abronius test method?
                                      165

-------
E 1383: FRESHWATER INVERTEBRATES




•    SPECIES-SPECIFIC ANNEXES




     Annex 1.   Hvalella azteca




     Annex 2.   Chironomus tentans




     Annex 3.   Chironomus riparius




     Annex 4.   Daphnia sp. and Ceriodaphnia sp.




     Annex 5.   Hexaaenia sp. (Main/Subcommittee)
                                166

-------
E 1383: FRESHWATER INVERTEBRATES





•    SPECIES-SPECIFIC ANNEXES (cont.)




            6.   Diporeia sp. (proposed; formerly Pontoporeia hoyi)




            7.   Lumbricyius sp. (proposed)




            8.   Oligochaeta (proposed;




            9.   Mollusks (proposed)
                                 167

-------
E 1383: FRESHWATER INVERTEBRATES





•    PROCEDURES:




     -Partial life cycle, static/flow-through, field contaminated/spiked sediment




     -Endpoints = survival, growth, reproduction




•    STATUS: approved standard




•    FUTURE PLANS:




     -Additional species-specific annexes




     -Hvalella azteca test method?
                                     168

-------
E1391: COLLECTION, STORAGE, CHARACTERIZATION, MANIPULATION
  . i    •        •      '-••.'••     •    "        .       '         „ '.   .
•    PROCEDURES:

     -Sediment collection, transport, storage, characterization, spiking, and dilution

     -Recommendations and Limitations

•   STATUS: approved standard

•   FUTURE PLANS: Test methods?
                                   169

-------
DESIGNING BIOLOGICAL TESTS WITH SEDIMENT





•    PROCEDURES:




     -Test type (e.g., whole sediment, pore water, elutriate)




     -Sample collection, handling, and manipulation




     -Test organisms and endpoints




     -Experimental design




     -Statistics, data interpretation, QA/QC




•    STATUS: Main/Subcommittee ballot
                                  170

-------
SEDIMENT TOXICITY TESTS WITH POLYCHAETES





•    PROCEDURES:




     -4 to 20-d test, juvenile/adult, field contaminated/spiked sediment





     -Endpoints = survival, growth




•   STATUS: Main/Subcommittee ballot
                                    171

-------
 BIOACCUMULATION BY BENTHIC INVERTEBRATES and





BIOACCUMULATION BY FISH





•    PROCEDURES:




     -10 to 28-d, static/flow-through, field contaminated/spiked sediment




     -Bioaccumulation potential vs. steady state




•    STATUS: Subcommittee ballots
                                  172

-------
RESEARCH NEEDS FOR STANDARD DEVELOPMENT





•   MULTI-SPECIES COMPARISONS




•   INTER-LABORATORY COMPARISONS





•   ABIOTIC FACTORS




•   LIFE HISTORY AND CHRONIC INDICATORS OF TOXICITY





•   SPIKING METHODS AND POSITIVE CONTROLS





•   DILUTION STUDIES AND MIXTURES




•   LABORATORY TO ]N STTU COMPARISONS
                               173

-------

-------
EPA Approaches for Biological Methods Standardization:
—    •  - perspective and Present Guidance
           ok, U.S. EPA Environmental Monitoring and Systems Laboratory -
                                                                            ' ฐ
I     Background - Standardization, of Biological Methods 1960 - 1989
      A    National Water Quality Network (NWQN)
      B'    Methods Development and Quality Assurance Research Lab
      C    Environmental Monitoring and Support Laboratory - Cincinnati
     ' D'.    Environmental Monitoring Systems Laboratory - Cincinnati
      E.    Biological Advisory Committee
      F!    Biological Methods Manual (1973)

 H.   Whole Effluent Toxicity Testing Manuals

      A.    Research Method to 304(H) CWA Approved Method (1981-1992)

 m.   Current Status of Biological Methods Standardization

       A.    EPA Biological Advisory Committee Charter

 IV.   Existing Agency (ORD, 1987) Guidance on Methods Standardization/Validation

       A.    Six Steps:                 N
                    1.  Determination of Method Requirements and DQO's

          ,         2.  Method Selection/Development

                    3.  Single-Laboratory Evaluation

                    4.  Confirmatory Testing

                    5. Interim Methods Description

                    6. 'Formal Collaborative (interlaboratory study)

  V.    New EMMC Workgroup on Biological Methods Integration
                                         175

-------
            OUTLINE OF ACTION TAKEN (1989-1991)
                       MAY 20, 1992
1.
2.
3.
4.
5.
6.
12/4/89
2/4/90
3/90
4/90
1/91
5/7/91
7.   5/27/91
8.   6/91
PUBLICATION OF PROPOSED RULE




PUBLIC COMMENT PERIOD CLOSED




PUBLIC COMMENTS SUMMARIZED




FOUR TECHNICAL SUBGROUPS  ORGANIZED




(AQUA.TOX., STAT, AMES TEST, VIRUSES)




RESPONSES TO PUBLIC COMMENTS COMPLETED




FINAL RULE TO RED BORDER REVIEW (OPPE,




OPTS, OGC, OW, REG. 3 & 65)  (NEW LAWYER,




REGAS, ASSIGNED)




RED BORDER REVIEW COMPLETE (OW AND




OGC NON-CONCUR)




FURTHER ACTION ON RULE PLACED ON HOLD




UNTIL TOXICiry TEST MANUALS ARE REVISED
                           176

-------
      BIOLOGICAL METHODS TO BE INCLUDED IN 304 (H)




                       MAY 20, 1992
PROPOSED ACTION:








1.   TOXICITY TEST METHODS



    A.   ACUTE AND CHRONIC TOXICITY TESTS FOR EFFLUENTS




         AND RECEIVING WATERS



    B.   ACUTE TOXICITY TESTS FOR DRILLING MUDS




     C.   AMES (MUTAGENICITY) TEST FOR EFFLUENTS, RECEIVING




         WATERS, AND SLUDGES.







 2.   METHODSFORCOLLECTION^ONCENTRATION^NUMERATION,




     AND IDENTIFICATION  OF VIRUSES IN SURFACE WATERS,




     WASTEWATERS, AND SLUDGES.








 3.   UPDATED REFERENCES FOR BACTERIOLOGICAL TESTS
                            177

-------
          INCLUSION OF TOXICITY TESTS IN 304 (H)




                       MAY 20, 1992           .








           RESULTS OF 1991 RED BORDER REVIEW:




              t    ' ,           -   •       •   .  '    -



1.   OPPE & OPTS* CONCURRED WITHOUT COMMENT









2.   REGIONS 3 & 5 CONCURRED WITH MINOR COMMENTS








3. .  OW CONCURRED CONTINGENT ON COMPLETION OF REVISED




    TOXICITY  TEST MANUALS , BEFORE  THE FINAL  RULE  IS




    PUBLISHED








4.   OGC NON-CONCURRED.  CONCURRENCE DEPENDS ON:




    A.  REVISION/APPROVAL OF FINAL RULE




    B.  REVISION/APPROVAL  OF   RESPONSES  TO  PUBLIC




        COMMENTS




    C  REVISION/APPROVAL/PUBLICATION OF MANUALS
* OPTS is .now the Off. Poll. Prevention & Toxics
                           178

-------
      STATUS OF INCLUSION OF TOXICITY TESTS IN 304 (H)


                          MAY 20, 1992




              ACTION STNCE 1991 RED BORDER REVIEW;



1.    9/91  -    REVISED ACUTE MANUAL SENT TO PRINTER   -    .
                                                  V
2.    9/91  -    NEW OGC LAWYER (SWEENEY) ASSIGNED

3.    1/92  -    CHRONIC MANUALS NEAR COMPLETION

4.    2/92  -    ACTION ON RULE RESTARTED (MEETING  IN DC WITH OGC,
       , -            •         i             •         |

              OWEC)

          -    REVISION OF COMMENTS AND RULE

              HARMONIZATION OF POLICY IN MANUALS, RESPONSE TO

          • ...  COMMENTS, AND FINAL RULE ,



      TARGET DATES FOR COMPLETION AND PUBLICATION OF RULE



5.    5/92  -     COMPLETION OF REVISED RESPONSE TO COMMENTS (RTC)

               AND FINAL RULE (FR)

6.    6/92  -     REVIEW OF RTC AND FR BY 304H WKGP

7.    7/92  -     RED BORDER REVIEW

8.    8/92  -     SUBMISSION OF RULE TO ADMINISTRATOR ,
   •/  >   - -           ••'   .      '         •     ••••''   '.
9.     12/92 -     PUBLICATION OF RULE IN FED REG


                          ,     179               .

-------
                  BIOLOGICAT ADVISORY COMMITTEE

The vision of the Biological Advisory Committee is to provide technical advice to the agency
on aU biological methods and related ecological issues.

MISSION OF THE COMMITTEE

1.  Review, comment, and assist Regions, ORD and Program Offices in standardizing and
evaluating EPA biological methods and indicators to be used by Regional and State programs.

2. Ensure that states develop logically consistent and ecologically meaningful biological criteria
that  facilitate  interstate, interregion  and  Environmental  Monitoring Assessment Programs
(EMAP) national comparisons.

3.  Exchange technical information and experience  in the collection, analyses, and use of
biological methods and indicators in assessing the effects of impacts on biological integrity.

4. Review, comment, and assist Regions and Program offices in developing agency biological
monitoring or biocriteria policy.                                            ,

5. Represent agency for ecological methods and other'related issues on National EPA and other
Federal committees (e.g., EPA Environmental Monitoring Management Council (EMMC) and
OMB Intergovernmental Task Force on Monitoring Water Quality).

Biological Advisory Committee (BAG) will be headed by a Chairperson from EMSL-Cincinnati,,
Deputy Chairperson from ORD or a Program Office.

Each subcommittee will have a chairperson and vice-chairperson, one of which is:

1.  an ORD or Program Office Representative, and
2.  a Regional Representative

The following subcommittees were agreed upon by the BAG during the FY91 meeting:

Toxicitv Assessment Subcommittee;

This subcommittee's mission is to address methods and other issues related to assessing acute
toxicity, chronic toxicity,  bioaccumulation, and organism physiological (biomarker level)
dysfunction due to contaminant in water, sediment and soil.

Ecological Assessment Subcommittee;

This subcommittee's mission will be to address methods and issues that measure ecosystem
health in aquatic and terrestrial systems.

The following addition of steering committees to the BAG Committee and its subcommittees was
recommended by the BAG Chairperson and Subcommittee Chairpersons during a November,
 1992 Society for Environmental Toxicology and Chemistry meeting:
                                         180

-------
RA.C Steering Committee;
Committee.

Tmricitv Assessment Su*ป™™™ittee Steering Committee;
 The mission of the steering committee will be to organize, prioritize, and expedite subcommittee
 activities.

 Ffnlogical Aซpssnient S"*ป™mniitteซ' Steering Committee;

 The mission of the steering committee will be to organize, prioritize, and expedite subcommittee
 activities.

                    REPRESENTED ON COMMITTEE
 1   A minimum of 1-2 Biologists from each Regional Environmental Services Division ^upon
 JecommTS  from  each  BSD and  Biologist  from Water  Management  and/or Waste
 Management).

 2.  A minimum of 1-2 Biologists from each ORD Laboratory.

 3.  A minimum of 1-2 Biologists from each of the following ProgramOffices:

        Office of Wastewater Enforcement and Permits
        Office of Science and Technology
        Office of Wetlands, Oceans and Watersheds
        Office of Solid Waste and Emergency Response
        Office of Toxic Substances
        Office of Pesticides

  COMMITTED  ACTIVITIES

  1. Quarterly conference calls:

         10 or less Regional Biologists plus 1-2 ORD Reps, 1-2 Hdqtrs Rep

  2. Information exchange in:

         Monthly surface water monitoring status report                 -
         Regional monthly reports circulated to regional biologists
         NETAC Newsletter    .                        ^
         Superfund monthly FORUM report                                   .        -
         EMSL - Bulletin computer board             ,
         EMAIL     ,
         FAX  ;   •        .     .   '        '    _.        _.••/-.   .';  '.

                                           181

-------
              EPA BIOLOGICAL ADVISORY COMMITTEE:
                      ORGANIZATION STRUCTURE
I.     BAG Chairperson and Deputy Chairperson
      EPA Biological Advisory Committee Steering Committee

      A.    BAG Chairperson and Deputy Chairperson
      B.    Subcommittee Chairpersons and Vice-Chairpersons
      EPA Biological Advisory Subcommittees

      A.    Toxicity Assessment Subcommittee

            1.     Chairperson, Vice-Chairperson, and Steering Committee

      B.    Toxicity Assessment Subcommittee Steering Committee

      C.    Ecological Assessment Subcommittee

            1.     Chairperson, Vice-Chairperson, and Steering Committee
            2.     Ecological Assessment Subcommittee Steering Committee
                                     182

-------
COMPOSITION OF EACH FPA BIOLOGICAL ADVISORY COMMITTEE
EPA Biological Advisory Committee Chairperson :<,

The  Chairperson of  the EPA  biological Advisory  Committee will be the Chief of the
Bioassessment & Ecotoxicology Branch.

FPA Biological Advisory Committee Deputy Chairperson:

The Deputy Chairperson of the EPA Biological Advisory Committee will be a willing ORD,
Regional or Program Office Representative appointed by the Chairperson of the EPA BAG.

EPA Biological Advisory Committee Steering Committee:    •   •    ,

The  Steering Committee will consist of the present and  past BAG Chairpersons and Deputy
Chairperson, Chairpersons and Vice-Chairpersons of the EPA BAG Subcommittees.
                                                                      COMMITTEE
gTTTON OF F, ACH EPA BIOL
 Toxicitv Assessment Subcommittee:

 The subcommittee will consist of BAG members willing to serve and a Chairperson nominated
 by anyone on the BAG, willing, to serve, and elected by the BAG.  The subcommittee will also
 include a Vice-Chairperson that is a willing ORD, Regional or Program Office Representative,
 nominated and elected by the members of the BAG Subcommittee.  Either the Subcommittee
 Chairperson or Vice-Chairperson should be from and ORD Laboratory.

 Toxicitv Assessment Subcommittee Steering Committee:

 The Subcommittee Steering  Committee will consist of a minimum  of 3 BAG members  (in
 addition to Subcommittee Chairperson and Vice-Chairperson) willing to serve and confirmed by
 Subcommittee Chairperson and Vice-Chairperson.  Steering committee should be composed of
 .a least 1 member from each of the following organizations: Regional Office, Headquarters
 Program Office, and an ORD representative.

 Ecological Assessment Subcommittee:

 The Subcommittee will consist of BAG members willing to serve and a Chairperson nominated
 by anyone on the BAG, willing to serve, and elected by the BAG. The subcommittee will also
 include a Vice-Chairperson that is a willing ORD, Regional or Program Office Representative
 nominated and elected by the Subcommittee members. Either the Subcommittee Chairperson
 or Vice-Chairperson should be from an ORD Laboratory.
                                         183

-------
Ecological Assessment Subcommittee Steering Committee:

A minimum of 3 BAG members (in addition to the Subcommittee Chairperson and Vice-
Chairperson)  willing to serve and  confirmed  by  Subcommittee  Chairperson  and Vice-
Chairperson.  Steering committee should be composed of a least 1 member from each of the
following  organizations: Regional  Office, Headquarters  Program Office  and  an ORD
Representative.
NOTE:  Tentatively the Committee and Subcommittee officers are filled by the following
individuals:
* EPA Biological Advisory Committee (BAG)

             Chairperson, Jim Lazorchak, ORD EMSL-Cincinnati
             Deputy Chairperson, Vacant

* BAG Steering Committee:

             Bill Peltier              ,
             Teresa Norberg-King
             Ron Preston
             Don Klemm
             Cornie Weber

* Toxlcity Assessment Subcommittee;

             Chairperson, Bill Peltier, Region 4 BSD
             Vice-Chairperson, Teresa Norberg-King, ORD ERL-Duluth

* Steering Committee:

             Joe Cummins, Region 10, BSD
             Cornie Weber, ORD EMSL-Cincinnati
             Margarete Heber,  HDQTS OW
                                       184

-------
* Toxicitv Assessment Subcommittee Members

            Peter Nolan, Region 1
            Robert Donaghy, Region 3                                       .
            Chick Steiner, Region 5
            Terry Hollister, Robert Vickery, P. Crocker, Region 6
            Mike Tucker, Region 7                 ;                     ,
            Loys Parrish, Glenn J. Rodriguez, Region 8
            Joe Cummins, Region 10
            Gary Chapman, ORD ERL-Newport
            Doug Middaugh, ORD ERL-Gulf Breeze
            Don Klemni, ORD EMSL - Newtown
            Phil Lewis, ORD EMSL - Newtown

      - Members signed up as of 6/91 BAG Meeting.  Others not attending 6/91 meeting can
also join, contact subcommittee chairperson.                                .

* Ecological Assessment Committee;

            Chairperson, Ron Preston, Region 3 BSD
            Co-Chairperson, Don Klemm, ORD EMSL-Cincinnati\

* Steering Committee;

            Jim Kurtenbach, Region 2, BSD
            Chris Faulkner, HDTS Wetlands, Watershed, Oceans
            George Gibbons, HDTS, Science and Technology

* Ecological Assessment Subcommittee Members

            Peter Nolan, Region 1
            Jim Kurtenbach, Region 2
            Jim Green, Region 3
            Del Hicks,  Jerry Stober, Hoke Howard, Region 4
            Wayne Davis, Thomas Simon, Region 5
            Evan Hornig, Region 6
             Gary E. Welker, Region 7
            Loys Parrish, Region 8
            Peter Husby, Region 9                    .           ,
             Gretchen Hayslip, Gerald Montgomery, Region 10
             Teresa Norberg-King, ORD ERL-Duluth
             Chris .Faulkner, OWOW AWD
             Margarete Heber, OST HESD
             Don Klemm, ORD EMSL - Newtown
             Brian Hill, ORD EMSL - Newtown    .
            Prank McCormick, ORD EMSL - Newtown
             Phil Lewis, ORD EMSL - Newtown

       - Members signed up as of 6/91 BAG Meeting.
       Others not attending 6/91 BAG meeting can also join, contact subcommittee Chairperson.
                                       185

-------
 TOXICITY
5NT SUBC<
CEFUNCTIOM
      The Subcommittee functions within the EPA EMSL-Cincinnati Biological Advisory
Committee (BAG) that provides guidance in the area of toxicity assessment.  The Subcommittee
is directed by a Chairperson with members representing EPA HQ Offices, ORD Laboratories
and Regions who volunteer to serve on the Subcommittee.

      The functions of the Subcommittee are as follows:

      *      Assist in the preparation, and coordination of toxicity test methods prepared for
             publication by the EPA for application in  freshwater,  marine and terrestrial
             ecosystems. EPA activities or programs impacted are as follows: NPDES whole
             effluent testing, sediment testing, toxicity reduction evaluation, dredge and fill,
             ocean disposal, EMAP,  CERCLA, RCRA, TSCA and FIFRA.

      *      Assess existing toxicity test methodologies (test condition, species, endpoints, and
             methods  of  data  analyses) that will impact  EPA National  and Regional
             implementation and enforcement of ecological programs.

      *'      Provide ORD laboratories and HQ programs  annually with a technical assistance
             needs list to support Regional toxicity assessment activities associated with the
             NPDES program, Water Quality Standards, and sediment criteria.

      *      Serve the Environmental Monitoring Council (EMMC) as a resource for review
             of toxicity testing methods proposed by other offices.  Provides information on
             Subcommittee activities relating to the development and use of new toxicity
             testing methods.

      *      Coordinate with  the Ecological Assessment  Subcommittee of the BAG on
             overlapping activities, such as biomarkers development and statistical analyses.

      *      Integrate toxicity biomarkers from the agency's strategic planning initiative into
             the activities of the Toxicity Assessment Subcommittee.

    .  *      Make available Subcommittee technical  expertise to State and EPA Regional
             offices  and HQ  programs for:  program, project, or report reviews; judicial,
             administrative, or legislative hearings; and adversarial proceedings.
                                        186

-------
                        ASSESSMENT SUBCOMMITTEE:
                   FTINPTTONAT. STATEMENT


The  Subcommittee  functions,  within EPA EMSL-Cincinnati Biological  Advisory
                                                                         The
                                 ,
Committee (BAG) that  provides  guidance  in the area of ecological  a^essmen^   The
Subcommittee is facilitated by a Chairperson with members representing  EPA HQ program
offices, ORD Laboratories and Regions who volunteer to serve on the Subcommittee.

      The functions of the Subcommittee are as follows:

      *      Assist in the preparation,  review  and coordination of ecological assessment
             methods prepared for publications by EPA for application in freshwater, marine
             and terrestrial ecosystems.  EPA  activities or programs impacted are as follows:
             water quality monitoring,  environmental indicators,  biological  criteria,  non-
             regionalization, EMAP, CERCLA, RCRA, TSCA and FIFRA.

       *     Evaluate  existing  ecological assessment  methodologies  (biosurveys,   field
             procedures, study design and  methods of data analyses)  that will impact EPA
             National and Regional activities utilizing ecological assessments.

       *     Provide ORD laboratories and HQ programs annually with a technical assistance
             needs list to support ecological  assessment activities associated with EPA and
             State environmental protection programs.

             Serve the Environmental Monitoring Management Council (EMMC) as a resource
             for review of ecological assessment activities proposed by other offices.  Provides
             information on Subcommittee activities relating to the development and  use of
             state-of-the-art ecological assessment methods.

       *      Coordinate with the Toxicity Testing Subcommittee of the BAG, on overlapping
              activities, such as biomarkers development and statistical analyses.

       *      Integrate ecological indicator goals from the agency's strategic planning initiative
              into the activities of the Ecological Assessment Subcommittee.

              Make available Subcommittee technical expertise to State and EPA Regional
              offices and HQ programs  for:   program, project of report reviews; judicial,
              administrative or legislative hearings; and adversarial proceedings.
                                          187

-------
            STANDARDIZATION OF BIOLOGICAL METHODS

1.     INTRODUCTION

1.1    BACKGROUND
       Field and laboratory "methods for monitoring  the status and trends  of the biological
integrity of aquatic communities and the quality of surface waters and effluents have played a
key role in Federal and State water pollution control programs for several decades. The current
Agency biological monitoring methods development and standardization program emerged from
activities of the National Water Quality (Monitoring) Network (NWQN) in Cincinnati in the late
1960's, later renamed successively the Methods Development and Quality Assurance Research
Laboratory (MDQARL), the Environmental Monitoring and Support Laboratory - Cincinnati,
and the Environmental Monitoring Systems Laboratory - Cincinnati.  Development, evaluation,
standardization and publication of biological (field) sampling and (laboratory) analysis methods,
first begun by  the NWQN in the late 1950's, has preceded uninterrupted to the present.

       The need for a formal monitoring methods development and standardization program was
recognized by  the federal water pollution control program (Federal Water Quality Agency) in
the late 1960's.   As a  result, an Agency chemical methods advisory group was  created to
recommend and assist NWQN in the standardization and publication of chemical and physical
monitoring methods  for water.  The first Agency manual of chemical and physical monitoring
methods was published in 1968.

       An Agency biological advisory committee was created by MDQARL in 1970, consisting
of representatives from  the regions, research laboratories, and headquarters program offices.
The members of the committee were selected to provide a cross-section of technical expertise
in biological monitoring and guidance on state, regional and headquarters program  office
requirements for aquatic biology data. At that time, the emphasis in biological monitoring in
the Agency water monitoring program was on the effects of discharges from publicly owned
sewage treatment plants on the structure and function of aquatic communities.

       Methods for  the  collection and analysis of biological samples  and interpretation of
biological  data were selected from the peer-reviewed literature and techniques then in regular
use by Agency regional  and research personnel and state programs.  Primary emphasis was
placed on taxonomic composition and standing crop.  Data on the identification and enumeration
of aquatic organisms were used to establish the status and trends of biological integrity in terms
of indicator organisms, the proportion of sensitive (clean water) and tolerant( polluted water)
organisms, and species  diversity indices.  The first Agency biological  monitoring methods
manual was published by  MDQARL in  1973 (USEPA,  1973).   The  Biological Advisory
Committee, established in 1970,  has continued to function to the present.
                                        188

-------
      In the mid-1970's, primary emphasis on biological monitoring in the Agency and states
began to shift from the biological integrity of ambient waters to the measurement of effluent
toxicity.  In response to the new Agency and state programs needs, EMSL, with the assistance
of the Biological Advisory Committee, published the first methods manual for monitoring the
acute toxicity of effluents and surface waters (USEPA, 1978) to aquatic organisms, now in its
fourth edition (USEPA, 1991), and methods manuals for the estimation of the chronic toxicity
of effluents and surface waters  to freshwater and marine organisms (USEPA, 1992a, 1992b),
based on methods developed by the  Environmental  Research  Laboratories at Duluth and
Narragansett, respectively.

             k              •               •                      •
1.2    CURRENT STATUS OF BIOLOGICAL METHODS STANDARDIZATION

       During this period of chemical, physical and biological methods development (21965 -to
the present), the Agency was also developing and continually strengthening its quality assurance
program, which rests heavily on the availability of standardized and validated methods.  The
development of the Agency's policy for a water quality-based approach to discharge permitting
(USEPA  1984) and  the subsequent move to place toxicity limits in discharge permits and to
include effluent toxicity  test in the list of "official" EPA (largely  chemical and  physical)
monitoring methods in Table I, 40 CFR Part 136, have led to questions related to biological
methods standardization, such as, "when (at what point) is a biological method considered to be
standardized or validated?" or,  "what process is involved in biological  methods standardization
and  validation?"  The agency currently lacks an official policy on methods standardization.


1.3    EXISTING   AGENCY   (ORD)   GUIDANCE   ON  METHODS
       STANDARDIZATION/VALIDATION

       The process of monitoring methods selection,' standardization and validation is essentially
similar for chemical, physical and biological methods.  A consensus document or "white paper"
on the subject  (USEPA, 1987) was prepared  by the staff of the Office of Acid  Deposition,
Environmental  Monitoring, and Quality Assurance (it has  since been renamed the Office of
Monitoring, Modeling and Quality Assurance, or OMMSQA), but the document and its contents
have not yet been "officially" endorsed by OMMSQA or the Agency, or promulgated as Agency
policy.                                                .

       Because of the detail and clarity of the 1987 document, it would be  senseless to repeat
 its contents in  full, here. However,  it would be advantageous to provide a summary of the
 process  described in the document, and to indicate  how the steps may differ, if at all, for
 biological methods.                                                               .
                                          189

-------
       The  six  major  steps are  described  below.   In the case  of biological methods
development/validation, the Biological Advisory Committee should be consulted during each
step.
1.
2.
tination of method requirements and data quality objective
       Determination

             - Provided by the program office

       Method selection/development

             - Potential user, such as program office and/or regions,  should  be actively
             involved.                             ,     ,

3.     Single-laboratory evaluation

             -  Includes  sensitivity  to  test method  variables   (ruggedness)  and  single
             laboratory/single operator precision.

4.     Confirmatory testing

             - Evaluation by several (minimum of three) independent labs.

5.     Interim method description _

             - Full method description, information on ruggedness,  mandatory and optional test
             conditions, guidance  on data analysis, single  laboratory precision, etc.   If
             endorsed by the Agency, it would now be considered a "standard" method.

6.     Formal collaborative (interlaboratory study)

             - Complete, acceptable,  data from a minimum of six labs.
                                         190

-------
                      METHODS STANDARDIZATION
BIBLIOGRAPHY:
1.   AOAC.   1984.   Report  of the Committee of Collaborative Interlaboratory Studies,
      Association of Official Analytical Chemists. J. Assoc. Anal. Chem. 67 (2)

2.   ASTM.  1979.  Standard practice for conducting an interlaboratory test program to
      determine the precision of test methods.  Annual Book of Standards,  14.02.  Water,
      Standard  E691-79.    American Society for Testing  and Materials,  Philadelphia,
      Pennsylvania.

3.  ASTM.  1981.  Standard practice of precision and accuracy of methods of Committee D-
      22 on Sampling and Analysis of Atmospheres.  Annual Book of Standards,  11.03.
      Standard  E3670-81.   American  Society for Testing and  Materials,  Philadelphia,
      Pennsylvania.                                              ,             l

4.  ASTM.   1985. -Standard practice of determination of precision and accuracy of methods
      of Committee D-19 on Water.  Annual Book of Standards, 11.01. Standard D2777-85.
      .American Society for Testing and Materials, Philadelphia, Pennsylvania. ',

5.  BatteUe  Columbus Laboratories.  1982.  Development of appropriate statistical techniques
      to compare analytical  methods across wastewaters.  Bishop,  T.F., F.E. Brydon, and
      B.C.  Dutter.  BatteUe  Columbus Laboratories, Columbus, Ohio.  USEPA Contract 68-
      03-2624.

6.   BatteUe Columbus Laboratories.  1985. Single laboratory validation protocol.  BatteUe
      Columbus Laboratories,  Columbus Ohio.   USEPA Contract  68-03-3224, Work
      assignment #1.                                                              .

7.   Glaser,  J.A., D.A. Foerst, G.D. McKee, S.A. Quave, and W.L.  Budde.  1981. Trace
      analyses for wastewater. Environ. Sci. Techn. 15(12): 1426-1435.

8.  USEPA. 1983. Guidelines and format for EMSL-Cincinnati methods. J.F. Kopp. U.S.
      Environmental Protection Agency,  Cincinnati, Ohio.  EPA/600/4-83/020.

9.   USEPA.  1983.   Guidelines for conducting single laboratory  evaluations of biological
      methods.   McKenzie,  W.,  and  T.  Olsson, m,  Bioassay  Systems Corporation.
      Environmental Monitoring Systems Laboratory, U.S. Environmental Protection Agency,
      Las Vegas, Nevada.  EPA/600/4-83/056.
                                        191

-------
10.  USEPA.  1984.   The development of data quality objectives.   Quality Assurance
      Management Staff, Office of Research and Development, U.S. Environmental Protection
      Agency, Washington, D.C.  Unpublished report.

11. USEPA. 1984.  Formal collaborative study design for water and wastes. P.W. Britton,
      ed. Environmental Monitoring and Support Laboratory, U.S. Environmental Protection
      Agency, Cincinnati,  Ohio.  EPA/600/4-83/020.

12. USEPA.  1987. Guidelines for selection and validation of USEPA'S measurement methods.
      Office of Acid Deposition, Environmental Monitoring, and Quality Assurance, Office of
      Research and Development, U.S. Environmental Protection Agency, Washington, D.C.
      Unpublished report.  57pp.

13.  USEPA.   1983. s Validation of testing/measurement methods.   L.R.   Williams.
      Environmental Monitoring Systems Laboratory, U.S.  Environmental Protection Agency,
      Las Vegas, Nevada.  EPA/600/X-83/060.
                                        192

-------
        STEPS IN BIOLOGICAL METHODS DEVELOPMENT.
              STANDARDISATION. AND VALIDATION
1.    Determination of Methods Requirements and DQO's


2.    Candidate Method Selection/Development



3.    Method Evaluation

           - Single Laboratory
           - Precision, Bias, Ruggedness



4.   Confirmatory Testing

           - Evaluation by 3 Labs

                                               \
5.   Interim Method Description

           - Now a Standard Method

"                   =' "         .          "       <

 6.    Formal Collaborative Study

           - Data from 6 Labs
                                  193

-------

-------
m
5t'
0

CD
O
r~ •
o
D
CAL METHODS
—
H
rn
Q
33
— r
— - '


^
z
p

r
IDATION






CO
OD

O
33
O

0
o
z'
o
m
0
z
m
H
X
0
Vp^
CO
CO
H

(O
(O
o
0)

m
5

CD
O
(LOGICAL ADVIS
O
33
•<

0
0
•. ^
^
o
1
CO
o

•p
m
PA APPROVED 1
^
m
H
^M
0
0
CO

m
5,
S
m
H
X

o
CO
PROCESS 1970
~i
1
^ป
(O
00
to
v*y

TI
O
^
_
m
H

O
O
S PROCESS 191
O)

- • 1 '
(O
, —*J
o
O
33

D
33
O
C
TJ
33
g
N
o
z
        -H
        m
      33
      O
      O
                         CD
                         O

                         5
                         g
                         o
                      cl
                      iง
                      O 03
                      ""-. 03
   m  m
      co
      CO
                         33

                         2
                         N
  .195

-------
RDIZATION
Is
1*
CO -=
O co
ORICAL METHO
Cornel iu
H-
co
n:
jm
<•
•i
•WE

CO
X
Q_
CO
Z)
i
ORING METHODS
^
^y
O
1
o
10
O)
- NWQN
x.
DC
NETWO
Y (MONITORING)
h—
~i
a
cc
LU
1-
1


T3
C
standardization,
field sampling
C CO
g eo
1.2
75.2
> .0
' ^^
+? O
ฃง
ฃ.2



methods
CO
"w
"co
c
CO
o
to
u.
,o


a
LJ.
(RING METHODS
O
H"
Z
O
n
CO
o
(D
CO
CD
LU
O
-^
\SSURAr
AND QUALITY )
h—
z
rELOPME
;>
LU
0






process:
^_
CO
E
AN


Q.
O
0
o
73
CO
•o
O
ฃ
ฎ
I *•
9- •
IAL - CHEMICAL;
METHODS
=3
" Z
CO
Q
O
^c
1-
LU
s

0
1
m
RING & SUPPORl
O

O
_i
L^
Z
LU
^

,z
o
1
CD
RING SYSTEMS L
Q

O
L^
LU
^K
LU
o  o ฃ'

                               1-
                               LLJ
a

Ll-
CO
CO
                                                           CO
                                                           CC
O   O
cc   cc
                                                                    LLI   LU
                                  196

-------
                   m
—
_
SSION: Provide techn
biological monitorini
program with emph
treatment plant dis
function of aquatic
o ฃ0 ซ•ป -•
0 3- CO _. g

i<3 ";1~
3 CD o rn 3
CCO 3 T3 TJ
w' "•' ,ป* w
3 > C
~ o *••• ••*
CD CD m O
CO ^. SJ. 3
' 3" CD CD
' ฎ IS "* 3
CD _ CD
CO 0 3 ~f
c w = ฐ
c -h ^
CD CO = o"
CD ซQ -i
ฃ0 $
3 03
Q.(Q





Guidance on State,
requirements for
& 3?'
•" CD
ฃ0 *ฃ.
ซ O
C^ ^^
ฃ0
cr-
0 50
o
CO tJ
^ -n
^^ ^k
O
0.0
10 S
•"*• --i
ฃ0 3
0
1^
o"
CD






Biological Monitoring
IU




















m
X
PERTISE: CROSS-SE
O
^^1
o
z

o
•n

H
m
O
I
z
o
'*"
/ m
X
T3
m
DO
— \
CO
N^ป
m


m
o
CO
3
o
— H
5'
CD
CO





















\
EPA Research Labs























EPA Regions





















DO
m
PRESENTATION:





















CO

0 Biological Advisor
*<

o
3

3
*+
CD
CD

I
^
D
O
DO
r~








LABORATORY -
O
ซ
Q
- ^•H
2.
Z
:>
^^•i
^^B




1








•*•
O
n
IS DEVELOPMENT &
O
ci
->
; n"
-
; ~2
• "^
| ^
CO
\#ป
CO
\Jr
DO
z
o
m
DO
m
CO
m
DO
o
, X


CD
O
n '
m
5





















                                  CO

                                  o
                                  5
                                  o
                                  m

                               Oj
                               o ^


                               CD m
                               lo
                                   3D
                                   g

                                   N
                                    O
197

-------
*
N
Q
DC
<
Q

1*
C/.J  0
CO  ^
Q  .J

li
LLJ  0)
•^  C
LLJ
_J

g
DC
O
H
CO
     co
     o
     4=
     CD
     .Q
               CO <0
              •2 ฃ
              H>
               ฃ n
               o o
O
UJ
=>
H   -r
.- 
             0)
             CO
             CO
 o I

 CO ^
f C
E ^

*^ CO
       —   o
               o
               CO
        ^   co w
          o
          O
          CO
          o
          h-
          ?   Q~  --ฃ

              ?     .ll
                      D) O1
O ฃ  .2
/-N —  ซ*-
            2 "o
            0
                      O
                      E
                     co
                     i-

                     LU
                     O
                     o
                                      ซ
                                      ซ

                                      "co

                                      o
                            0)

                            4-ซ
                            CO
                            0)
                            CO
                            o
                            co
                            4->
                            CO
                            CO
                            o
                            DC
                            H
                            LJJ
                            2
                            O
                            CQ
                                      CO
                                      CD
                                 CO    O

                                 o   ?
—   c
  .   o
 o   '"5
U—   k.

 I   1

•3
 C
 CD
                                .
                            CO  4-<
                                          O
                                          I^M
                                          *-

                                          O
                                O
                                O
                    O

                    z:
c
CD
                                          CD

                                          Q.
                                          i_
                                          CD
                                      o
                                      0
                         .2   ฐ

                         ง   I
                         ?   ฐ
                         O   i
                                         O
     >   Q.
     i   s
     DC   O
     S   I
                                               CO
                                               CD
                                               O
                                          o ฑ:
                                          •^ co
                                          CO Jr
                                          i_ CD
                                          g >
                                          E -o
                                               __
                                               c co
                                               O ซ-
                                              ~ _CD

                                               00
                                              -51=
                                              0.2
                                                        DC
                                                        UJ
                                          O
                                          DC
CO
CD
CO

"co
c
CO

CO
*~
ca
               i  2  •ฃ

              LLJ ซ?   O
              H      O

              SE •—   c
              CD      o
                      O
                     ^0)

                     "o
                     O
                                                                    o
                                                                    0.
                                                           CO
                                                           D)
                                                      CO
                                                           
-------

e
1
CO
•-f
5T
0
o
CD"
*ir
tA












%
B>
g.
— K
OJ
ftj
21
Q%
C
ft%
•Mi*

O
0
3 •
C!j
O
^^™
^H*
3,
ฃ
QJ

CD
™*

CO
CO
^•^
1
m
r~
-Cincinnat

c
cr
co"
,3-
CD
CO
3
fi>
3
,-c
05
, *^™
6










m m CD
g x o
1 s s"
(Q -l Q.
3"
tQ
^
CD
3
O
*<
D
>
CO
c
a
Jป
3
CD












*+•
V> O
0 3
modificat
and capa
cro
=: 3
~ CO
*< ^
0 ฎ

m co
> Ol
CD 0
CO O
o'S
3 3
jh% cO
— 3
r~
a>
cr
CO


,
93
Q.
CO
C,
5T
o
CD
DJ
i-f
CD
-T
CO
















CD
3"
O
ct
-t
measuring

CD
DJ
o
c
CD
0
X
^jr
ซ<

o

CD
^^^

__
•C
CD
3
CO
I
3 m
Gi ^
)*S ซ3h
If2
fi> .
•Cincinnati
1 on:
**
O
o
3
cp_
E"
CO
CD
cr
CD


•o

cr
co"
17
CD
CO

m
*
CD
^^h
CD
CO
CD
Q.
O
3
CO
H
O
o
3
3
<-ป•
J-C
O
CD
CO
m
i
CO
en
.
__
i.

CO






3
o
o
3'
vertebrate!
w
fi>
3
^^i
ftj
^
cr
S"
CO
" a









CO
01
3- '
o rn
0. 0 33
co 3 r-
o"
fi>
o
c
c?
r^>
o
X
cv
^4>
ป<
^.
31
_,,
co"


"








o
C
c
o
CD"
CO
CO
CD
%•/
T5
0)
•
•D
C
co"
3-
CD
CO

m
5

.^
CD
TJ
O
                                   Q
                                        D- CO
                                        O  ^
co
          199

-------
S STANDARDIZAT
o
STORICAL EPA METHO
31
ซBI
4H
_/•
•
•Hi
Eงfii
9ซiii
ll
Jal
RONIC TOXICITY
X
EMPHASIS SHIFTS TO C
CO
0
o
X
••H
LU
del
i

CO
o
CO
o>


Cincinnati Publishes:
i
_i
CO
S
LU

Q
DC
O

1

CO
CO
O)
Tiethods."
mtm
15
C
"o
c
O
i
CO
LU
u.
O
<*—
to
E
^
o
TJ
C
CO

CO
CD
^)
2
3



j—
ater Establishes Policy or
ฃ
"o
CD
.0
H—
0

1

CO
O>
Permits

CO
LU
Q
CL
Z
O
+*
JC.
0
CO
O
0.
a
CO
•o
CD
CO
CO
m
1
>>
+*

"co
d
^

T3
0)
CO
CO
X2
I
it^
"co
: =J
O"
1_
o
*-
CO
I_
o
H—
*-
c
o>
h
•o i
tz
5ง
Is
CO X
o
co *-
"E
O.

!r

• •
• c
O
nati - Publishes manual
-Cincin
_i
CO
LU

i

1C
CO
o>
*E
o
&••
o
D)
"co
E
to
(D
O
H—
CO
•a
o
"5
E


ฃ
i
ซ
mmm

CO

CO
3
15
5
^^
f effluents and receiving
ater organisms."
1
rl
o *-
._ <*-
X
5 ฐ






CO
4^
I_
O
Q.
0>
b.
ฃ
LU
cz
•ซ *"^
uth peer review methods
EPA biologists working
nee Division laboratories
3 0 C
Q CD
1 CO CO
_i ,2
DC "•ฃ eg
LU IE •*•*
JQ Jr
5 ซ c
o a. E
ซc
^^
•ooo
Q) ' .1=
CO ^ >
CO c C
CQ CO LU

200

-------
^  m CD
O  "0 8s
X  > CO
7T   O
=' -i Q.
-. O
3  -i
    .
   co  m
      33
'<• 3
S-?-
3—  w ^
   CO Ji>
0)  TJ CD
3  0> &)
^•0-3
 s?o  a
 <& CO  ป-ป-

 p" -*• -'i"   =•*
                   CO
                   =r.
                   
-------
Process
O
00
REQU
(-
o
<

DC
LU

fe
         LU
         _J

         O
         CO
         h-
         o
         LU
         CO
              CO
18
CO
LU
DC
pursuan
              2
              0.
     4->

   m ซ5
   Q; o
   ^5 m^m f—
   •_ ซfcซ ^
   ^ ~ O
   Ti ** •—
   t? k. *-

   C CD CO

   •~ o o
                _
              0 -=
              .
              D>
       "co > g
       •= c S1
       co co rc
             si-
             iz: 4_> T3
                   CD

CO

CO
              co
     CD
    2

    '3
     D)
ซP CL

C '> ซ-
CO O O
                .
                II?
How
     c
     o
                OT  ฃ ฐ ^,
                •—    ป'J3 2
.- ซ Q
S =
co a aj
O QL-—
SS CO
OL
Q.












c
eo
*-
CO
eo
N—
^T^
^,
CD
0

E
0)
Q,
CO
LU
O
a.


CO
"co
TJ
4i^
C
3
H—

0)

0>

•-ป
15
15
3
o-
k.
O


-------
ro
GO
c--'
D-
,_!.
0
CL
o"
CD
O
— K
O
5"

0

I














•3
C
ฃ.
03
<3
W*
0
CL
D"
03
0
CL
0
3
"D

' g;
o'
03
"*1
CL

CD
^>
O

o
o
3
3
0
3
ft~m


O 03
2.3
0 "
5" 0"
3 &
0
O
CL
03
01


"TJ"
C
03
0
03










1


as
&3 X
03 MM*
ty methods
tics




_














CD
CD
O
CO
o
^^
I
03
ubgroups f
o
i
0
CL
j_|-
O
ฃป
CL
Q.
0
O3
03

0
O
.3
3
0
3
03
CD
0
-n
C
g;
o"
Comments
o
o
3
•D
0
t-f
0














         ro
         CD
         CO
         CD
       c m
       t*
       o-o
         Q
       .- 03

       -*• O
       CO 3
       O>
         I-*

       & ง

       -i 2
       CD ^
       .Q *<

       ฎ S-
       03 CD
       O (Q
       O
       3
       3
       CD
       =J
       i-i-
       CA
                   O
 203

-------
s
O O

X CO


eง
o^ o:

co co
w d
LLI <
O 13

0 Z
cc <
Q. S
00
1 Border review
T3
0
CC
J_
o
**—

"0
0
*^
"i
-Q
13
CO
fl5
3
1
LL
• •
O)
O)




i
-ฃJ
/IS
E.
i_
^
o
c
0
0
1
c
o
c
o
O
O



0
E
E
0
o
_o
Zi
=3
Q.
O

CO
0
CO
c
o
Q.
CO
0
i_

0
+-ป
CO
c
o
[to
0
*i^



s manuals to be
0

"5
0

•ป
J2
CO
13
C
CO
E

_0







_0
L_
en
L_
Q
M—
0
_Q
0
IMIBI
^2
Q.











1

TJ
0
i_
3
O
c
0
0
0









•o
0
x:
.S2
15
3
Q.
CO
CO
CO
E
c


"5
L-
- _
o
-a
0
•^ '









c
o


•o
0
concurr
__
CO
13
CO
E
0^^
w-d
ฎ 0
JZ
>ป.S2
•rr ^
^J "™t
• ^^B ^aJ
X CL
0
*- T3
C
ฎ CO
13
o ^

0
GC



CM
CO

JI
*j

>
manuals

o
o
o
CM
•
i- w
^ *-*
o c
0
w E
t E
=3 0
gง
O M_
o o
1
5 *ฐ
O 0
C O)
co
> Q.
O


• •
CM
CJ5
O)
   CO
   3
   CO

 0 S
 w eo
 o c

ฃ ฐ
   ฐ t.

.6.0 ฐ

*~< ^
+- mZ-


 &*ง

 sr-
 0

 w -^ 0 52

 C 0 ฐ •-
 c 4ii co *-
 0 to    ~
E •ฃ ^r co
CO
c =.'
        CO
O O CO JD
O O O =J
     •^ co
*-•*-• *r c
CO O -C 3

0 c S
c   0 ซ^
                                                 -.
                                                0
  — fl>  CO

   g co.2
     o  •*-
     JZ  CO
                                                O o    CO
                                                CO fl) *- *-
                                                  •S O co

                                               Q
                                               cc

                                               o
                                 204

-------

m

I

Z
o

m
5
CD
O
r-
OGICA
r-

S
m
H
X
0
O-
CO
>
z
o
z
0
o
o
33
CO

<ฃ
O
m
O
'z

H
m
O
X
z
o
>
r-
co
CO
,c
m
CO

0
m
>
r;
z
0

ง
-I
I
CO
5
'z
O
33
ฃ3


33
m
m
:>
ซ•

o
o

^^
2.
m
•y
- ^~
H
>
CO
CO
CO
H
^_^
33
m
D
0
Z
CO
ซ•

O
33
O
"
3ฐ
"D
33
O
o
33
.^•'
<ฃ
CO
>
o
D
c
H
m
CO






















. o

 >

 D
     5*  TO
     m  33


     3  i
     O  O
     D  m
     CO

     >  m
     z  o
     D  X
        Z
     33  n
     PS


     m  >
     o  |

     m  rt
     O
     O
             CO

             >
             0
                    co -*
         m
         CO
           m
         O O
         Q

         ง 3
         2 m
      Co Q
      co m
        z
        o
z
D
           O

           Z
CO

o

o
D

O
^
z
o
X
m
13
33
m
CO
m
z
S
O
z



















ป

CO
c
CO
o
o

_
-5
-i,
m
m
CO
•n
c
z
o
-i
o
z .
r~
CO
^
m
S
m
z
—i
CO
o
~n

CO
c
CD
O
O

s
H
H
m
m
^
z
D

T3
^D
33
O
m
o
CO
$.
^
CO
CO
o
z
, ••
o
H
m
CO
0
33
Q
Z
N

^
6
z







H
m
m
33
Ci
il'' •
.0
o
Sfc
^
H'
H
m
m
D
33
m
:>
^_
•D
O
I ,
>
33
H
m
33
>
O

CO

0
33
m
<
m
^
m
D

CO
CO
-*
CD
^
^'
i

33
. m^J
m
o
X
>
33
-1
m
33
m
o


















Z!H
zm
QO
02'
if
coS
m
z S
coo
/ISORY COMM
US COMMITTE
mq
H
m
m
O







m
5'
03
O
OGICAL ADVI
1990s
o>
O
xซ
X
o
o
s
H
H
m
m
                   205

-------
LU
LU
H
t
•5
.OGICAL ADVISORY COMf
1990s
__1
O


n i
UJ
J
i
n
t^^M
^Ifl
CO
Of
r (REGIONS, ORD, & PROGI
CO
CO
ff\
CO
!_..
-^
\C DUTIES
REVIEW, COMMEr
•^ •
CQ t-
THODS
LU
^fmm
. ISSUES DEALING WITH
.UATING EPA BIOLOGICAL ti
— ซ —I
— m
"Z.
oง
, ^
AND VOTE ON 1
STANDARDIZING








.
AND INDICATORS


_j
^-*
CONSISTENT & ECOLOGIC/
CL
O
_j
UJ
UJ
Q
ENSURE STATES
cvj

CO
CRITERIA THAT FACILITATE
AL, AND EMAP NATIONAL
J-O
o 7n
TK UJ
ฐcc
7> HI
MEANINGFUL BIC
INTERSTATE, INT









COMPARISONS.

"CO
UJ
ORMATION AND EXPERIENC
AND INDICATORS.
u_co
2 Q
-o
_jX
o LU
•^ *^
EXCHANGE TECHI
ON BIOLOGICAL
CO


1ST AGENCY BIOLOGICAL
CO
CO


K"
^^
REVIEW, COMMEh
•


;RIA POLICY.
UJ
E
o
o
CO

MONITORING OR

ELATED
cc
IOLOGICAL METHODS AND
CO
2Z
O
>
,-^
REPRESENT AGEr
id
0
LU
AND OTHER FEDERAL AG
ITFMWQ
CL .
LU O
_I ^
_^ป ซ(E
< LU
O r
ISSUES ON NATh
COMMITTEES, i.e

206

-------
1
o
r-
m
NFORCEMEI
z
H

Z
<
m
CO
H
O

H
O
o
^ ^
m

H
m
33


0000
Tl T| T| T|
00
m m
OO

SOLID WAS
POLLUTION
H
TJ m
33
m >
ป m
zg

> ฐ
z ^
o S
^^
s^
m
33
~
-T-
CO ฑ
TJ
c
2
''•
. o
. CO
0 S
•n m
0 O
m z
o w
1
10
WASTE WAT E
33
m
Tl
O
33
O
x f'
m
Sป
rn
H
*>
TJ
m
TJ

~ •
H
CO
m
CO
r~
r-
ff\
CO
VEGAS

















m
CO
i-
g
g
1

















m m
33 33
I~ I"
1 1
Q b
CO
r- 33
mmm ^
^%
LLIS
BREEZE

















m
TJ
r~
i
z
33
•n
Ju
5>
X3ANNSETT


-














s ง
r~
i
o
r~
-J
I ;.-.






-









f
O
33
Q
Z
N
• 5
O'
Z,
CO
33
m
TJ
33
m
CO
m
z
I
—1
m
o
0
z
03

0








•.
TJ
33
O
D
•MB
33
t>
,M OFFICES
•*r*
~ ^
D
_
1
O''.
CO
m
i

o
6
P™
ง^^
Q
CO
33 CO
D











.
33
0
s
33
m
Q
O
Z-

p.
i
rv
CO
m
                                           O
                                           m
                                           CO
                                                   m

                                                   5
                                                   ro
                                                   O

                                                   O
                                                   g
                                                   o
                                                   >


                                                •
-------
 LU
 HI
 h-
 I-
O
O

DC
O
CO  co
O
O
3
O
m
LLI
LU
CC
z>

O
Z>
CC
F-
co
<
z:
O
           LU
           LU
   O
   CO
   CC
   LU
2 0.
OCC
CO
< I-
X =>
O Q.
   LU
   O
             CC
             O
             CQ
o
O
O
                     CC
                     LU
                     LU
                     h-
                     co
                     CQ
                              LU
                              CO
                              S1"
                              L1I m
                              coil
         38
                                           o
                                           LU
                              •z.
                              Q
                              CO
                              CC
co
DC<
HIT
                0>
                                           LU
                                           CO
                                           CO
                                           >
                                           I-
                      O
                      o
                      CQ
                                        O
                                        CO
                                        CC
                coE
                oc.<
                LUX
                S=ฐ
                CC  i
                                                  0>
                                             LU
                                             LU
o
o
m
D
CO
o
       CC
       LU
       LU

       CO
                                                          LU
                                                          LU
                                  o
                                  o
                                  m
                                  3
                                  co
                                            EE
                                            CC
                                            LU
                                            LU

                                            CO
                                         208

-------
!  I
=r
o
0.
CO

CO

2.
CD
O
CO
a.
D

        "D
        C
        3D
        m
CL
CD
O
O
CD
CO
CO
3
o
I-  5
•ฃ  a
o  ซ
    3'
fi?   Q.
3
Q.  -?
    ca
    c
D.  S-
fi}  O
           03
CO

c
to
z
Q
           -n
           3D
           m
                m
                H
                I
                O
                o
                05
                     O
                     pj

                     Q

                     Z
Z
H
m
z
-i


I
to

o

m
                     co
0
3D
03
O
r~
0
Q
O
^p^

m
•H
X
0
a
CO
                      co
                          m
                          m
                          m
                          (O
                          CD
D
C
o
m
                              m
           •g  O
           33  3D

           5  CO
                              m
                       3- m
                       H  o

                       m  O
                       H  z
                       I  .
                       O  >

                       S' ^
                           s
                           O
                           •n
                                 O
                                 3D
                                 O

                                 O
                                 O
                                 m
                                 z
                                 a>
                                 c
                                 CO
                                 o
                                 o
                                  m
                                  z
                                  H

                                  O
                                  33
                                  H
                                  m


                                  5
                                  "0
                                  m
                                  3D
                                               3D
                                               m
                                            (A
                                            2
                                            5>
                             So
                                               3D
                                            g
                                            N
                                  ^     =!-5
                                  3=  •   -0 >
                                            35  m
                                            c  o
                                            g  z

                                            II
                                                o
                                                a
                                                CO
                    209

-------
CO
Q
O


s
O

UJ
13  <
03  N

UJ
CO
UJ
oc
a.
    1
CO
          CO
          Q.
          UJ


          CO


          X

          CO
          CO
        V  UJ
              UJ
              Q

              z>
              C5

              U.
              O

              >
              CC
              CO
                    CC
                    oc
                    CL
                       UJ
mo


LU <

II
OC5
OC UJ
CL OC
LU
                    O
                    
           UJ 111
           CO O

           o
           o
                                    UJ
                                c\
            CO

            UJ
              I .^^k.

         U_ CD CO
CO
         UJ

         CD

         _J
          i
         LU
         J
         CD


         55


         co"
                                     oc
                                  9=0
~ UJ
ฃ*S'
— • LU
> QCD
H0<5
-* ^^
งsง
O CL <
CO O_l
TT 1 1
m m m
yj j _i
                                                      coco
                                        210

-------
  O)
  •


	-TV
CD
  O
i;
51
  m
                      m
                      DO
                      m
                      O
                      D

                      O
                      m
                      CO
                      O
                      DO
O
o
z
•n
DO


I
O
DO
 H
 m
 CO

 z
 o
                               CO
                               c
DO


O
-n

O
o
m
m
CO

o
O
                                       m
                                       co
COMPLETE ACCEP
DATA FROM A Mils
OF SIX LABS
•"•• —4
s ">
C CD
	 ^^tm
ENDORSED BY AG
•STANDARD" METH
O m
o z
o
CO
m
ir
CD
i
•o
DO
m
g

CO
o
GUIDANCE ON DAT-
ANALYSES
3>


MANDATORY & OP"
TEST CONDITIONS
—1
O

FULL METHOD
RUGGEDNESS TES"
—i .
m

EVALUATION OF A
MINIMUM OF 3 LAI
TO
CO

                                C
                                m
                                D
                                       ฐ
                                      N


                                      5 1
                                      0>
                                      C O
                                      5Z


                                      II
                                        O
                                        a
                                        CO
                    211

-------
DC
O
                                             LU
                                             Se
LU
h-
it
1- CC
GICAL ME1
WORKG
O
O
CQ
o
LU
mi

••••^^^^^n
21
3B
aBB
&SSI
agj^m
LU
Z
QC
O
^^
LL.
O
O
CQ
O
o
LU
0
• cc
LU
CO
III
LU
=>
o
LU
QC
CM
O

CO
Q
O
X
LU
O
CD
O
_J
O
CQ
Z
O
Cu
O
CD

DC
O
^




CO
CC
X
o
1
o
o
o
•1-
cc
o
LU
1-

_I
CQ
CM
o>


BORATORY
_i
ROM REGIONAI
LL.

LU
Z
O





LU
O-
LL.
LL.
Q
^sr
ROM PROGRAK
LU

LU
Z
0



' •

CC
X
o
1
o
o
Q
CC
T
HAIRMAN IS Tl
O

CQ




o
CC
o
1
a
CO
o
EPRESENTING
QC





OMPEND
O
QC
III
LU
O
CD
CD
z
IS CONSIDERII
O

LU
CO
o>
s_
OTTRS
CQ
3 STARTED
w*
Q
O
X
DLOGICAL MET
^^
m~~

LU
O



O
ง
COMPEND
LU
CO
LU
1-
CL

O
t-
z
LU
h-




f
Z
o
6:
QC
CD
LU
z

CO
o
o
fZ
LU
IO
                    00
                                       212

-------
EMMC Methods Format
William Telliard, U.S. EPA Office of Science and Technology


I.           Scope and Application - Tabular format whenever possible

             A.     Analyte list
             B.     CAS numbers                          .
             C.     Matrices                                                    •  •/=
             D.     Method sensitivity (expressed as mass and as concentration with a specific
                    sample size)
             E.     Data quality objectives

n.          Summary of Method                                           "
          .                                      \                          .    •
DDL.         Definitions

IV.         Interferences

V.          Safety

             A.     Above and beyond good laboratory practices
             B.     Disclaimer Statement (look at ASTM disclaimer),
             C.     Special precautions
             D.     Specific toxicity of target analytes or reagents
             E.     Not appropriate for general safety statements

 VI.         Equipment and Supplies

 VH.        Reagents and Standards
                           i
 Vm.       Sample Collection, Preservation and Storage

             A.    Provides information on sample collection, preservation,  shipment and
                    storage conditions.                 -
              B.     If holding times are  exceeded, data may have changed and should  be
                    flagged for data user's attention.

 IX.          Quality Control

              A.     This section provides a summary of the QC requirements of the method.
                                         213

-------
X.           Calibration and Standardization

             A.     Should include  calibration steps  that  are  not  followed  daily; daily
                    calibration steps will be included in the procedure section.

XI.          Procedure

XII.         Data Analysis and Calculations

Xm.        Method Performance

             A.     A precision/bias statement should be incorporated in the section, including
                    detection limits, and source/limitations of data.

XIV.        Pollution Prevention

             A.     Cite good laboratory practices for pollution prevention.

XV.         Waste Management

             A.     Cite how waste and samples are properly disposed.

XVI.        References

             A.     Source documents                                               ,
             B.     Publications.

XVn.        Tables, Diagrams, Flowcharts, and Validation Data

             A.     Location of these items will be left to the judgement of the individual
                    work group.
(Finalized as a result of January 24, 1992, balloting by Members of EMMC Methods Integration
Panel and Work Group Tri-chairs.)
                                        214

-------
              m
              O

              O

             (3
              &>


              5
              3
215

-------
c
0

E
Q.


"0










w
^>
c
05
Q.
O
ฃ







CO
Service
0
o
c
03
CO
CO
>
15
O








c

+- ป
03
O)
0
ฃ
CO
"D
O
oC
A

ฃ
3 .
• ^™
"D
C
0'

Q.
E
0
o
CO
T3
O
ฃ
0
T3
0
4-ป
03
E
o
4-J
tt
0
Q
c

• ^BBI
-M
03
'•\rf
"5
'D)
0
DC
63
CO
•o
O
0
^
15
•^
15
c
•

c
.2

*+-*
<0BW
03
4— '

T3
O
O
O
to
O
03
zi
03
C
•2'
03
-Z
                   IB
216

-------
,NJ

6
o


v '•




-
- •' •

%p
3
• •
• . '

D > co
D ^ ฃ
o CD ^
^ W' CD
<•. Q) o
0 ^ B:
*-!'!'
-, 7T O
ฐ r^ 2.
Cu^ 7T
— • " ""* -^
^ o Q
c ™^
•D 0
i- C" .
d^H
/D


I


ฃt
CD
^•M
^^
0
Q.
' Q)"'
O
vT
o
o
c


S
o
~
o
o
c

(A
'





g
o
o
&
CO
o

3
(A
a
E
&}
j*
o
3


                                                     217

-------
(ft
B
o
(6
             U)
             C'

             to
             0)
             U)


             "5
             CD

               &
          o  a>
         E.E
          c/>
          3
             fl)
          CO  >

         0)  il
                        CD
                        0
                        ,b   0)
                             CD
                             0
                            -C

                             0
 CD
 C
 o   >
 S   CD
 0

 C
 O
 o

!5
"o
 CD

 O)
 C
 o
                        CO
O

ฃ
        <
o      <
TO  CO   0
D^  LLI   O
C  
-------
Si
ro

o
o
'•'<ฃ.
0
Q.
03
>-*•
CD
C7
CD
CO
CD-











,
CO
i
o
o
3
0"
Q.
CD
r-+
C .
"D
0
5.
CQ
0
CD
o
^ w
3
0
5"
O
Q_
5'
CD
CD
0
0
•—
0
X,
0)"
. *^*
•
CQ
3
0
^^*
IT
O
Q.
CO

O
O
CO
CL
0
3
6'

>ป "h
ฃ ฐ
S-. 3
0 si
0 ^
•D^
.I-*"
0
|
D'
0
o
CQ
•<
m
|
^^
o
Q.
3
^^^^ป -
^^^r




• •
> Q
CQ 0
0 5"
0 CD
Approac
Methods
/-s 1 **" .^^—
g o 1 _ 31
Miv ^^^^ • • ^^^^^^
30
S co"
o
a.
— h
ฐ
3
Q)
m
|^^ ' •
^
i
3-
flT
05
:
5"
ฃ"
3 H.
rt o
(D W
(Q
3
Mil
O
3









  tn
                                   219

-------
 3
 O  (ft
 ^  O
O
•••••  	

S-2


t






"d
3
O
5

Q
ซ
tt\


CO
c
CD
• •
M—
"D
C
CD
CO
.
'x
o
LIZ
•D
co 0)
1 1
O .'C
"•P O
CO —
0? O
CO
CD
0)
T3
C
—
"o

o

v>
o
^3
o
E
c
0
c
o
"Z.













c
-.2
[E
"a)
Q
   Q.
         05
                              Uj

                              t
m


o
o

cvi
in
<
                  220

-------
o
o
                       en
                       0
                       a-
                        i

                       a
                       CD
                       CD

                       o

                       CD
                       D)
                       o
                       0)
o_

a



CD
                                            O
                                            •t
 o
 c
WQ

^ C
OlC


3
                            221

-------
   1C
   V*
   c
   o
   0
S-g
   a.
a
3
O

O


o
CO

5
0)
              D)
              CD
     CD
c

.2
"•+-ป
CD
o
                    CO
                    C
                    o
a.o

II
O Mz
s s
p 0
                              I4J
                        o
                        o
                  222

-------
O C G
0  D  D
O


O
—h

3
0
O
Q.
0
O

3
o
0
   o
   D
   0
   O  O
   Q)  Q)
   O
   D

O  O
   9- D
      o
      0
      c
   W   0

       3
       0

       r-+
       O)
          I   I
Q.
Q)
f-t
o'
13
      O
      O
      0
      Q.
      0
      05
             0
   9L
   51
   0)
   o
   Z5

   CD
   C

   CL
   0
             0
             05
            0)

            o'
O5
05
C
0
05
                    O
                    i
          ""  ฃ
          5-1
          2,  *•

          I'?
          W  O
              S
              T3
          O

             223

-------
 W

**
    o
 0)
    CD
•o
 ^
*  %
 0 CQ
 O J.
 C  O
 E  c
 S(5
 0>  (0
Q.  >
            CO
            c
            _o
            "•*-ป
            CD
            O
            J^
            "o
            0
            Q.
            CO

            "D
            O,
            0

            E
            J0

            JD
            CD
            0
            Q
                0
                CD
                JD

                0
                CD '13
                CO CO
                CO
                0
                O
                0
                •- o
                0 0
                00
cS O
O
                0.0
                CO Q

                JD -Q
                .a 0
                X"™" •—
                  CO
                ^ 0
                      ^ -5
                      CO T3
                      CO 0
                      0 O
                      o o
                      0
                      c
                        CO
                      0
                      o
                      c
                      CD
                        0
                        0
      0 CO
      Q. 0

      0 ••' =
        CT
       ^ 0
      "O *~
      O >
                      0

      0
                      CD O
                                UJ
                                •I?
                  224

-------
Ul
to
6
o
 *
  m
   33
   CD
CD CD
                           m
                      o
                      <"
                        <  o'
                         03
                         ง i"
                         o a.
                         w  -w
                         C  Q.
                         0-91
                         r   *"*
                            CD
                         D  Q)
                         O  O)
                         0  0
                         O)  =

                            D
CDCQ

0>g

D m
2L
<
^
o'
                        H
                o
                3
                                              mm
                        O
                        3


                        3
                    O   fl)
                    a.  -
                    V)

                    3"
                    a.
                    (D
                    X
                          225

-------
      x
c  -u
1   =
=  T3
 C   0)
Lil




      o
 (B   S
      O

CO
 5
O
                  0
                     CO
       c
       O
O)  CO  O
C  O M3
'^  o "ฃ

2  S  E
c  >  J=5
O  CO
    c
       —
 0  Q. o
LLJ  X

   -ง
    c
                                    
-------
Freshwater Sediment Toxictiy Assays:
Necessary and Desirable Attributes
G. Allen Burton, Jr., Wright State University, Department of Biological Sciences - Dayton, OH
I.    What's Available?


H.    Assay Strengths/Weaknesses


ffl.   Assay Requirements

      A.    Necessary attributes (from a scientific perspective)
      B.    Desirable attributes (from a regulatory/program/project perspective)

         • .   .              •                 -   '                   , ; >
IV.   Examples of Assay Evaluations


V.    Preliminary Recommendations
                                        227

-------

-------
      Toxicity Testing



usefulness improving rapidly



- method refinement



- short-term chronic assays



- toxicant interactions simplified



- reduced uncertainty
              229

-------
           Selected Freshwater Sediment
 Organism Group

 Amphibians

 Fish



 Zooplankton
Toxicity Tests

 Test Species
 Response Measures

 Embryo-larval survival, Terata  Xenopus laevis
 Embryo-larval survival, Length
 Weight, Terata
 Survival, Reproduction
 Benthic Invertebrates   Survival, Size, Reproduction,
                      Molting, Emergence, Avoidance
Microbial

Phytoplankton

Macrophytes
Benthic Indigenous
   Communities
Luminescence (Microtox™)

Cell number, 14C uptake

Frond number, chlorophyll,
biomass, root and shoot length,
peroxidase

Structure indices, functional
indices, chlorophyll, respiration,
enzyme activities
 Pimephales promdas
 Oncornynchus mykiss
 Oryzins latipes

 Daphnia magna
 Ceriodaphnia dubia
 Brachionus sp.
 Colpidium campylum

 Panagrdlus redivivus
 Caenorhabditis elegans
 Tubifex tubifex
 Stylodrihts heringianus
 Pristina Iddyi
 Lumbriculus variegatus
 Hyaldla azteca
 Diporeia sp.
 Gamrnarus pulex
 Gammarus fasciatus
 Corbicula fluminea
Anodonta imbedllis
 Chironomus tentans
 Chironomus riparius
Hexagenia  limbata
Hexagenia  bilieata

Photobacterium phosphorewn

Selenastrum capricormttum

Lemna sp.
Hydrilla verticillata
Bacteria
Protozoan
Periphyton
Phytoplankton
Macroinvertebrate
                                      230

-------
            Optimal Tpxicity Assay Issues
Validation                    Resources   ,
  Relevance                       Organism availability
  Sensitivity/discriminatory         Laboratory resources
  Exposure design                 Expertise
  Response dynamics               Expense, time
                     Standardization
                       Methods
                       QA/QC criteria
                       Adequate database
                          231

-------
Selection of Optimal Assessment Endpoints




    Project objectives



    Site characteristics        .



    Available methods
   •*


    Key components represented
                   232

-------
Assay Sensitivity:  effect vs. control or reference
                Realistic protection of study ecosystem      . .
               (i.e., nationwide, ecoregion, site-specific)
                         Relevance
                         Validity
                         Significance
                             233

-------
                                      5  S
       I
       I
       0.
       •8
       co
       en

 •o
  O
  3
  (A
  O
  3
  
       03

       W
rr
O
       (B
       CD

       2T
        i

       CL

       g


       I
       M
                                                       CO 00 CO O CO 2 O
                                                                   la
                                          234  '

-------
Factors affecting sensitivity

 • Measured response
 • Organism type
 • Life stage
 • Health
  • Test design (e.g., exposure period)
  • Sample manipulation
              235

-------
 LLI
 CO
 z
 o
 Q.
 CO
 LU
 DC
   SENSITIVITY
     UNDESIRABLE
THRESHOLD
             M
     [CONTAMINANT]
     DESIRABLE
LLI
CO
Z
O
CL
CO
LU
EC
THRESHOLD
     [CONTAMINANT]
                 (Ross et al. 1951)
         236

-------
DISCRIMINATION
     UNDESIRABLE
UJ
en
z
o
O.
CO
111
CC
THRESHOLD
     [CONTAMINANT]
      DESIRABLE
UJ
en
z
O
Q.
cn
UJ
cc
THRESHOLD
     [CONTAMINANT]
                  (Ross et al. 1991)
         237

-------
       Necessary Attributes



•  Sensitive (responsive)

•  Discriminates (discerns degree of contamination)

•  Relevant to ecosystem/study objectives
   (Species and exposure design)

•  Validity (field verified, few false +/-)
                238

-------
        Desirable Attributes
                                          /
A. Agency Specific:
   •  Comprehensive indicator                 '
   •  Reliable
   •  Resource requirements
   •  Standardized

B. Additional Program/Project Specific:
   •  Uniqueness (non-redundant)
   •  Confirmatory (welght-of-evidence)
   •  Significance (ecosystem, commercial, societal)
                .  239

-------
What Makes An Assay Relevant?



   Consider the:

       Test ecosystems' characteristics

       Sample manipulation artifacts

       Organisms' route of exposure(s)

       Organisms' ecological niche

       Measured response sensitivity
      . (e.g., mortality vs. reproduction)

       Organism stress
                . 240

-------
For example:







     Trout * Carp



     Hyalella * Aquatic worms (e.g., Tubifex)




     Elutriate * Solid phase



     Pore - Solid phase ?



     Benthos - Nonbenthos ?



     Lab response i- In situ response ?
              241

-------
Total Quality Assurance







 • Study design



 • Sample collection/manipulation



 • Exposure design



 • Assay performance criteria
           242

-------
      O

      a

      w
      C
             p

             3
CD
•o
•i
O
a
c:
o
a

3

en
a
                    Q
P

3
rฐ
•p

Cft
C
             ffi
c;
•-a
                                 •p

                                  (A
                    Q
             p

             p.

             •1
             CD
O
a
                                                                   rฐ
                                                                  •b
                           0    2.
                           w    ฃ1
                           —    o
                                                                   ca
to
O
CD

O
O

O
                                       1
                                                                                       0
                                                                   0
                                                                   c/5
                                         243

-------
                                                                                  C3
                                                                                  O
                                                                                  •^H

                                                                                  O
                                                                                  e
                                                                                  f-l
                                                                                  u
                                                                                  in
                                                                                  ฉ
                                                                                  0>
                                                                                  c
                                                                                  O
                                                                                  (D
CD
                     ^^-   • •   f—•  V • "*  ^^   ^^
                     ^   g   o   g  S  ฃ  5
                          ^^   '• *   *^  ป**   *C  "P^^
^  ^   ^  Z.   w
P.  fc>   U  t>   ^
                          q
                         ffi
•05-
                                                u
              D)
                                   (0
     ft  >  CO
                       >•<  X~X >"**  _. t
                       D  ~   o  t:
              ฃ  r:   6  ~  .ฃ
                                       •=  'ฃ  ป  ii
ft   -  z+
fi\  CU
                              •q
                              o
                                       ft
                                       oi
                  g
                  ci
                           Ul

                           G
                 '6
                 •J
                              O   "'  O   U  O   U
                                                              CO
                                    >  6
&   S   ^  r,
ฃ  ^   .  Q
                                    ft
                                   OH"
                                          244

-------
                         Survival
 CD

 •ง•"
 O'
M

cw
(D
H
cn
6
(B
rr


05
                X\\\\\\\\\\\\\\\\\\\\\
            x\\\\\\\\\\\\\\\\\\\\\\\\\va
                                                             (D

                                                          _ C/3
                                                          3  2
                                                          ฐ    -
                                                             W
i-1
\D
>*
O
                                            245

-------
 o
75
 m
 (D
0)


0)

O

QC
 o •>
I- E
   CD
    CD
   Q.
          !
              (0

              1
              O
              a
                      T5
                      0)
                      o
                      o
ฃi
ฃ c
fig
f I
.ง
i
kk
•^
fl)
                                    o
                                    o
                                    o
                                    01
                                    o
                                    GO
                                    o
                                    n
                                    o
                                    CM
                                    O c
                                    (0
                                      i
                   246

-------
I
w
CD
D
en
en
O
•-t
 O
 I—^
 o
ffi
Q

•>j

a
en
            Q

                       a
                       a
_   a
                  en
                  C
                            P
                            a
                            CA
           <   CD
           h—   ซ-i
           <   Q
           a   Q
                      s
                      I—>.
                      o
p
a

>j

a
•-ซ
CD
t3
•-t
O
a
                                                  00
                                                  tr

                                                  en
                                                  Q
ffi
                                                        a
                                                        en
in    tn
      O


      lซ^
      O

      O.

      en
      C
                                                       Q
                
-------
  G
  O
  O
  G
 "i— I

  6
 F-4
 u
Q
 co
 G
 
g   ?
•o    o
o    ฃl
t~t    D&
a   ^
   ~    >
                O
C
0
 o   i-
 >    ^
 o
 e
•o
o

XJ
ffi
                   a
                  cu
                             en
     •q
     K
                                  o
           u
           c/i
                CO


                A
                CO
2
a
ฃ
m

ฃ
b
JQ

6
00
TT

6
CO
      g
      m

     •o
                      fl    a
                     Q    a:
                                      248

-------
        rฐ
        br
         i
        to
        CO
        a
        en
        a
Q
a
w
p

O
                e


                Q
tJ

a


o


tr
                        CO

            o

            ฃ•
            o
            *^
            Q
                       a
                       3
Oo
cr

CO
C
•i
                                       CO
CQ

3
    •K
    Q


    a

    (A
                                Q  Q   C  Q
                                ป—•  •—  9  i—
CO
a
(Q
a
CO

O
 B

 p
                                       H-i-
                                       S
                                      CQ

                                       O
                                       O
                                       3
                                       t5
                                       O
                                       13
                                       0
                                       2_
                                       c/5"
                      249

-------
          Principal Components Factor Analysis:
                   ARCS Assay Comparison
Factor 1           Factor 2            Factor 3           Factor 4

P. .promelas (wt.)    D. magna (survival)   Hexagenia (survival)  Hexagenia (molting)

Ii azteca (survival)  C dubia (survival)    C dubia (young)

D. magna (young)

P. promelas fterata)


                          Variance explained

      40%             23%             13%                n%
                               250

-------
               Summary Ranking
              (Ingersoll et al., 1992)
                Protectiveness  Similarity   Sum
                    ;"        -
H. azteca 14-D         1            3       4

H. azteca 28-D         2            5       7

C. riparius 14-D       3            1       4

Microtox              4            2.6

D. magna             5            49

C. tentans 10-D        6            6       12
                       251

-------
        Sediment Toxicity Sensitivity Comparisons



•  Twelve studies by Ankley, Burton, Cairns, Giesy, Hoke, Ross,-et al.

•  Comparisons of 3 to 20 assays/study



Most sensitive assays

   Group A:  Hyalella azteca 7-14 d survival

             Chironomus riparius 7 d survival

             Daphrtla magna 2-7 d survival

   Group B:  Ceriodaphriia dubia 7 d reproduction

             Pimephales promelas 7 d larval growth

             Chironomus tentans 10 d growth

   A = Most sensitive in at least 2 studies
   B = Second and third in sensitivity in at least 3 studies
                               252

-------
               Test Battery Recommendations

                         Burton et al.  Giesv-Hoke   flC    ITC
Daphnia magna (48 h)
/              (7d)
          •-•'•-           .'(or)
Ceriodapnnia dubia (7 d)        •
Hyalella azteca (7-14d)          •
              (28 d)         (or), '                          *

Chironomus riparius (10 d)      •                            (or)
                            (or)
Chironomus tentans (10 d)       •     .        •            ,   *
                            (or)                          (or)
Hexaeenia limbata (10 d)        •                             *
Microtox
             J         \
Selenastrum capricornutum

Algal fractionation bioassay

Ames
                               253

-------
              General Purpose Short List






 I.  a. Microtox (screen in tandem)




    b. Ceriodaphnia dubia or Daphnia magna (3 brood)




    c. Hyalella azteca, Chironomus tentans or C. riparus.



         or Hexagenia limbata (7-14 day)






 n.  a. Pimephales promelas (early life stage)




    b. Selenastnim capricornutum






ffl.  Other assays in tandem with above
                     254

-------
Extreme
   Slight
     I
 Pristine
U
Need to Integrate Assessment Tools
                                                  Burton & Scott  1992)
                               255

-------
              Conclusions






• Consider all assessment issues '




• Test multiple, relevant species




• Test multiple trophic levels




• Validate laboratory responses




• Use proven methods




• Expect site variance
                256

-------
Midge Whole Sediment Bioassays
John P. Giesy andJody A. Kubitz, Michigan State University - East Lansing, MI
                 . midge Chironomus tentans has been effectively used as a bioassay organism
to predict the toxicity of sediments to benthic organisms. This organism is easy to culture and
maintain and  standardized protocols  exist to produce a  continuous  supply  of known-aged
individuals.  The organisms are hardy and  easy to manipulate in  bioassays.   Growth of ^
tentans is a sensitive measure of response, which gives  good discrimination power among
^edim^nts   Studies have been conducted to  calibrate the response of the survival and growth
studies to community structure, and some information on the relative sensitivity of the midges
to chemicals is available.

        There are several issues that need to be addressed if C, tentans assays are to be adopted
for routine use.  Theses include: more studies of the relative sensitivity of the  midges to
chemicals; more comparisons between laboratory and field  studies; investigations of the genetic
drift of laboratory cultures; and comparisons of partial life cycle tests with whole life tests for
toxicants of several modes of action.
                                           257

-------

-------
Q
CD-
*<
? CO
E CO
''•••-" O

N"
,
CD
CO
CO
O
3
co
c:
CO.
CD
Pฐ
CD
CQ'
^" .



CD
CO
O)
•^
Q
O

"3
•
CO
3"
13
Q.
Q.
N"
CD


259 .


•.=5" o
Q. ง
<" i:
• s'i
c c
So
^— C
^**
C/5
c
"O
^•1^
1
o
D
1
CQ
CD




O
^^*ซ
> 3
CL^
< O
& ^
zj 3
i-*- S^
JD O)
CQ -H.
CD CD
O) ^3
53"
0)

'


-------

•
CO
c:
s —
ง CO
-fS CD
CO O)
^ +s
E C
0 ^
c ^
^^^^fc ^^^^^^p
ci ^
"^
o

CO
E'
•2
CO
o
o
CD
D)
ง
O
O
O -!-•
=" ฃ
CO CD
3 C
to '*-
C O
'


O
• ^^^1B
informat
CO
o
"o>
o
o
o
"x
o
-_
o
1
260


.
c •
CO
Q.
CO
\CD
ฎ.
^•••^
.Q
ฃ
O
CD
Q.
^
O
CO
•
^^^^M



•D
CD
D)
CO
CO
•D
_>ป
"55
CO
CD
o
1
>*
^^^ m
CO
JZ
•
•^H
CM
O>
O>
 N

"S
 3
o3
>ป
(0
a
5

-------
Q

CD*
0)
X


O1


J*.


tD

ro
    Q
    O
     i
    en
    CD
    g.

    3'
    CD
    CD

    ft
    C/3
    hf:   O

    3
    CD
CD


CD

i—t
C/3
     03


     Q.
     Q.

     N"
     CD
     Q.
          03    CD
               O)

               g;

               CD
                    CD

CD O

03" 91

CD C/3

Q-CQ-
           o =
           o o
           O 03

           I"
           3 CD
                     CA
             ^
           c
           o
                     CD
         Q.

         O)"
         O
              FT   3
                             03
                             g
                              CD
                              o
         O
                              CD
                              C/

Q.P
                  o

                  3
              &  ฃ,
             CQ  CO
              CD
                                  C/>  CD

                                       CO
                    261

-------
CO
c:

c S>
m cp
CO
   CO
 O "^
 C  ^3
    CO


65
  TJ
  O
= O
CO D)

  CO
  0

g>.i
  H-J
  0
  E

O CO
       o
       (D
          CD
sens
       >>
       ii
       s  ฐ
              CO
       CD
                0
                O
                CO
                CO
               ฃ w
               0 0
               E-Q
                CO
                QL
       CD
       CO
              O 0
                0
                Q
                CO
                CO
                0
                     0
                     CO
                     2 co
                     CO '
             EO
             o
             O CO

               o
             0  ซ-
             co  E
             CO
             CO  CD
             0?CO
              262
                              CM
                              O>
                              O>
N

in
3
                              <#
                              (0
                              Q>

                              O

-------
o
en
S ฎ
i i
N- '• ' • MMI
^
CD
•••8 ง
M ง
2,
Q.
•• . • . • c
tn




. \




•
•O
O
CD
••••I
D
21
6~
CO
^*
. ^^
•^
CO
d!".
o
g




•
5- c/>
o g
MB* • /NN
C ฐ-
S P"
, -3
CD
X
T3
CO.
3"
CD
Q-
•^•B
O
CO
CO
2,

CQ'
o
— *

•
CQ
CD
D
CD
o"
Q.
5?-
"3
Q.
^^ ^
CO
y* ^
U
I

1
5^
CJ
CO
co'
ฃZT.
o
•13




Disadvan

rP^
CQ
CD
f/\
—






. •
o
hironomu
C/)

Cn
ijjj
^3
55"
0)

•

263

-------
CO
C
c:
mus
sues
no

s
CO
4-J
GO

O
            0
            co
           "D
   GO
_-  0
       O  O
                   O)
                   ..  o
                      • M^^^H
                   GO
o    -^

                         264
                                  03
                                  CO
                      CD
                      E
                                  (D
                                  GO
                                  GO
                                  E
                     E
                      "     ^
                            T3
                             
                                     (0
                                     (D

                                     O

-------
o
5'
0)
< •
* ZT CO
c ^^*% dปr*
5". y CD
N^ ^V •*""'
••O — -
S- 03
N> ฃ+L
13
2,
CD
Q.
3"
CD
1— K
1
' "' -' *•<
CD
CO
;•••

•
-3.
o
^^^
3
o
CD
0
D
N"
03
^^T™ '
m**m •
O
D
C/3
CD
Q.
3"
CD
D^
i
•
CD
O)


•
03
0

IZfL
o
,, 13
i
v^
0
cn
O








•
•v;-'--h-
O
'; f.
i— h
O
c
(Q
1
0
O


'-




•
<
5s
0

0
oT
o
0
3
0
13 .
i— i-
0
O)
D
l^H^^^
O
.






o
^ \^
ง•
^^^^^ '• ^^^ • '
O) O
C ^
CD ง
^ ^
- 1
= cB"
^3
rT*"1
w
^j
CO
265

-------


.
CO
C
45
c
CD
"**'
CO
_^ w
ป**J
5
o
c:

•fc
O





CO
"O
(D
CD
Z
HMBM
* *•ป
^MB
O
CO
CD
CO
^^ ^
CD
DC






c
_o
"CO
.Q

"CO
O

[o
1

1
JD
_CC
CD
O
E

c
g
•"*
: accumL
*t—
O
.
o

>•
^i
O
"x
o
CD
1Z3
CO
CD
O
E
CO
to
CD
CD
CD
O
"1
O

CO

CO
/-%
LJ-
'o

g
.g
fv*
CO
O
•
>


CD
••iM^k
C3)
CO
^ ^^
^^^^^M
C/D
Q3

CD
J^^^J
• •M^H
CO
C
CD
CO
to
O
E




c
g
j^^^j
_^^^j
CO
^2
• •K^H
_^_^J
CO
O
o
V—
o
_CC
1
0
.E












CM
0)
O)
N
3
_**
266

-------
Q
5'
w

8ฐ

c
o;

N"
CO
CO
ro
          T5
           CD

        CL9:
        0)  Q
0

^' O
        CD  CD
        a s
           O
           O
           o
           13
           CO

           w"
           (Q
           O
           O
           Q.
0)


0)
                CD g-
                &S
                   a
                   CD


                   CQ
                   CD
   O
   O
           CQ   i-
                CL
                05
                              CD
                           c
               o
                           CD S

                           Sฃ
                           O CD
            CD  c/)
            Q.-
                            O  ฑf.
                               CO
                              CO

                              O
                    CD
                    Q.

                    0)
                    D
                    Q.
                              CD

                   08
                   O
                   Z5  O
                   a1.3
                        c
                           CO
                           o
                                        CD
                                   C/)

                                        CO
                    267

-------

-------
Desirable and Necessary Attributes for Freshwater Sediment Toxicity  Tests:
Hyaletla azieca        ,                                    ,
Chris Ingersoll, U.S. Fish and Wildlife Service, NFCR - Columbia, MO

                                      *
I.     Objectives

       A.    Life history of HyjteHa azteca
       B.    Culture and test methods
       C.    Research needs for standard development                     .


H.    HvaleUa azteca Life History

       A.    Species:  HyaMla azteca (Saussure; talitrid amphipod)
       B.    Habitat:  lakes, ponds, streams
       C.    Distribution:, North America and Caribbean
       D.    Salinity:  Euryhaline; fresh .water up to about 22 g/L, culture 10 to 15 g/L
       E.    Life stages: Immature (1st 5 instars), juvenile (6th & 7th instar), adult (8th instar
             and older; about 35 d at 20 degrees C)                              ,
       F.    Growth:  Indeterminant; male larger than female; male enlarged gnathopods
       G.    Feeding: Omnivore; bacteria and algae < 65 um
       H.    Behavior: Epibenthic, burrow in sediment w/o vegetation


in.   HvaleUa azteca Culture Methods

       A -  Flow: Static renewal, or flow through
       B.    Temperature:  20 to 25 degrees C
       C.    Light: 16:8 photoperiod; 50 to 100 ft. candles
       D.    Chamber:  1 L to 100 L
       E.    Age of animals:  Known age vs. mixed age
       F.    Water quality:  Soft water (ERL-Duluth) vs. hardjvater (ERL-Corvalis) strain
       G.    Aeration: Moderate
       H.    Feeding:  Maple leaves. Tetramin.  Rabbit Chowj diatoms
       I.    Substrate:  Maple leaves. Mitex screen, cotton gauze, 3-M base web plastic
                                         269

-------
IV.    Hvalelia azteca Test-Methods

       A.    Flow:  Static, renewal, flow through
       B.    Temperature:  2Q to 25 degrees C
       C.    Light:  16:8 photoperiod; 25 to-50 ft. candles
       D.    Chamber:  25 mL, 1 L.  up to 100 L
       E.    Sediment ration:  1:1 to  .1:4 ratio sediment:  water
       F.    Age of animals:  Known age (0 to 7 d, 7 to 14 d) vs. mixed age (size about 7 to
             14 d)
       G.    No.  animals: 5 to 20/beaker; 4 to 5 replicates/treatment
       H.    Duration: 7 d, 10 d. 14 d, 28 d
       I.     Endpoints: Survival, length, weight, sexual maturation (males), young production
       J.     Water  Quality: Soft water (ERL-Duluth) vs. hard water (ERL-Corvallis) strain
       K.    Aeration: None or moderate
       L.    Feeding:  None, Rabbit Chow, YCT, maple leaves, Tetramin
       M.    Acceptability:  survival (80%). length, weight
       N.    Particle size: low sensitivity (with sufficient food)?
       O.    NH3 and H2S: low to moderate sensitivity?
       P.    Sediment contact:  Mayflies = Midges >  Amphipods >  Daphnids
       Q.    Sensitivity:  Daphnids > Amphipods = Mayflies > Midges
       R.    Reliability:  Amphipods  = Daphnids  > Midges  >  Mayflies
V.     Research Needs for Standardization of Hvalelia azteca

       A.    Culture and testing: know-age and' feeding
       B.    Culture and testing: reconstituted water
       C.    Reconstituted sediment
       D.    Abiotic factors
       E.    Reference toxicants
       F.    Species and strain sensitivity
       G.    Inter-laboratory comparisons
       H.    Life history and chronic indicators of toxicity
       I.     Spiking methods and positive controls
       J.     Dilution studies and mixtures
       K.    Laboratory to in situ comparisons
                                         270

-------
OBJECTIVES




•     Life history of Hvalella azteca




•     Culture and test Methods




•     Research Needs for Standard Development
                                 271.

-------
HYALELLA AZTECA LIFE HISTORY
Species:




Habitat:




Distribution:




Salinity:
Hvalella azteca (Saussure; talitrid amphipod)
lakes, ponds, streams
North America and Caribbean
Euryhaline; fresh water up to about 22 g/L
                Culture 10 to 15 g/L
                               272

-------
HYALELLA AZTECA LIFE HISTORY (cont.)
Life stages:
Growth:




Feeding:




Behavior:
Immature (1st 5 instars)                 •   •   " '




Juvenile (6th and 7th instar)




Adult (8th instar and older; about 35 d at 20ฐC)




Indeterminant; male larger than female; male enlarged gnathopods




Omnivore; bacteria and algae < 65 urn




Epibenthic; burrow in sediment w/o vegetation
                                  273

-------
HYALELLA AZTECA CULTURE METHODS
Flow:

Temperature:

Light:

Chamber:
Static, Renewal, or flow through

20to25ฐC
              /
16:8 photpperiod; 50 to 100 ft. candles

1 L to 100 L
                          274

-------
HYALELLA AZTECA CULTURE METHODS (cont.)

Age of Animals:          Known age vs. mixed age

Water Quality:
Aeration:

Feeding:

Substrate:
Soft water (ERL-Duluth) vs. hard water (ERL-
Corvallis) strain

Moderate

Maole leaves. Tetramin, Rabbit Chow, diatoms

Maple   leaves.   Nitex   screen,   cotton
gauze, 3-M base web plastic
                                 275

-------
HYALELLA AZTECA TEST METHODS
Flow:



Temperature:



Light:



Chamber:



Sediment ratio:



Age of animals:








No. animals:
Static. Renewal, flow through
20to25ฐC
16:8 photoperiod; 25 to 50 ft. candles



25 ml, 1_L, up to 100 L
1:1 to 1:4 ratio sediment: water
Known age (0 to 7 d, 7 to 14 d)
vs. mixed age (size about 7 to 14 d)
5 to ZQ/beaker; 4 to 5 replicates/treatment
                                2-76

-------
HYALELLA AZTECA TEST METHODS (cont.)
Duration:
Aeration:
Feeding:
7d. 10d.14d, 28 d
Endpoints:       SyiylvaJ, length, weight, sexual maturation (males), young production




Water Quality:    Soft water (ERL-Duluth) vs. hard water (ERL-Corvallis) strain
None or moderate
None, Rabbit Chow, YCT, maple leaves, Tetramin
Acceptability:    Survival (80%). length, weight
                                    277

-------
HYALELLA AZTECA TEST METHODS (cont.)





Particle size:     Low sensitivity (with sufficient food)?




NH3 and H2S:     Low to moderate sensitivity?




Sediment contact: Mayflies = Midges > Amphipods >  Daphnids




Sensitivity:       Daphnids > Amphipods = Mayflies  > Midges




Reliability:       Amphipods = Daphnids > Midges > Mayflies
                                   278

-------
RESEARCH NEEDS FOR STANDARDIZATION OF HYALELLA AZTECA


•   CULTURE AND TESTING: KNOWN-AGE AND FEEDING

•   CULTURE AND TESTING: RECONSTITUTED WATER
                         /     .,                  i  , •
•   RECONSTITUTED SEDIMENT

•   ABIOTIC FACTORS

•   REFERENCE TOXICANTS             '

•   SPECIES AND STRAIN SENSITIVITY
                             279

-------
RESEARCH NEEDS FOR STANDARDIZATION OF HYALELLA AZTECA (cont).
     INTER-LABORATORY COMPARISONS
     LIFE HISTORY AND CHRONIC INDICATORS OF TOXICITY
    SPIKING METHODS AND POSITIVE CONTROLS
     DILUTION STUDIES AND MIXTURES
     LABORATORY TO IN SITU COMPARISONS
                             280

-------
Discussion of Desirable and Necessary Attributes for Marine  and Estuarine
Sediment  Toxicity  Tests,  and the  Use of Ampelisca  abdita. Rhepoxvnius
abronius. Leptocheirus plumulosus. and Eohaustorius estuarius in Marine and
Estuarine Sediments
Richard C. Swam, U.S. EPA Environmental Research Laboratory - Pacific Division
I.     Description of Acute Amphipod Sediment Toxicity Test


n.    Research and Regulatory Applications


m.   Limitations and Advantages
 ,   "  • '  •"   '       •      •   '  •'      '   '•       '   ;  '  v •-  '

IV.   Necessary and Desirable Attributes of Acute Sediment Toxicity Tests

      A.    Species selection
             1.     Relative sensitivity (field and lexicological data)
                   a.     Other species
                   b.     Size
                   c.     Sex
             2.     Ecological importance/relevance
             3.     Economic importance         ,
             4.     Habitat     -,.'.'•.'
             5.     Substrate relation
                   a.     Pelagic
                   b.     Epibenthic
                   c.     Infaunal - tube dwelling
                   d.     Infaunal - free burrowing
             6.     Availability
                  , a.     Field collection   ,
                   b.     Culture method
             7.     Laboratory compatibility   .
             8.     Zoogeography
             9.     Compatibility with bioaccumulation/chronic tests
                                       281,

-------
       B.    Method development and standardization
            . 1.     Written standard method
             2.     Sediment toxicity database
                    a.      Field sediment
                    b.      Spiked sediment
             3.     Control responses
                    a.      QA/QC - collection/culture sediment
                    b.      QA/QC - reference toxicant
                    c.      Experimental - negative
                    d.      Field - reference sediment
             4.     Statistical power
             5.     Tolerance limits of species/method
                    a.      Sediment grain size
                    b.      Temperature
                    c.      Salinity
                    d.      Sediment organic enrichment
                    e.      Ammonia
                    f.      Seasonality
             6.     Sediment collection/processing/storage
             7.     Field validation
             8.     Interlaboratory comparison
V.     Amphipod Species

       A.    Rhepoxynius abrom'us
       B.    Ampelisca abdita
       C.    Eohaustorius estuarius
       D.    Leptocheirus plumulosus
VI.    Chronic Test Methods


   .   Other Species
                                        282

-------
     TABLE  I—Precision of the benthic bioassay in relation
                 to sample size and replication.*

              Number of Amphipods per Replicate

              10                             20
No. of
Replicates
2
4
6
8
10
12
14
16
18
20
6
6.80
2.66
1,94
1.60
1.38
1.25
1.14
1.05
0.98
0.93
6/c
71.6
28.0
20.4
16.8
14.5
13.2
12.0
11.0
10.3
9.8
No. of
Replicates

2
3
4
5
6
7
8
9
10
( •
a

8.55
4.44
3.35
2.80
2.45
2.20
2.02
1.89
1.76
d/c

45.0
23.4
17.6
14.7
12.9
11.6
10.6
10.0
9.3
  a6 is the difference between two survival means for which the bio-
assay is 75% certain of detecting statistical significance (P <  0.05)
[76]. 6/c expresses the precision estimate as a percent of the normal
control survival (c = 19.0 for n = 20; c = 9.5 forซ = 10).
                             283

-------
FIELD VALIDATION
PARAMETER
BENTHIC BIOMASS
BENTHIC DENSITY
SPECIES RICHNESS
AMPHIPOD DENSITY
RHEPOXYNIUS DENSITY
EH
BOD
OIL/GREASE
CADMIUM
CHROMIUM
DEHP
DDE
Toxic
SITES
9.4
450
21
0,8
0
-54
13
15
29
670
16
5
NON -Toxic
SITES
50.3
2000
70
33
23
+223
4
4
11
370
4
3
      284

-------
1.  Written Protocol

   Biology of Test Species.
     . all1 species selection factors

   Limitations of Method
      environmental factors (grain size, salinity, etc.)
      variability/statistical power       ,   .
      field relevance

   Logistics                             >
      exposure chamber                   ,
      duration         • ,  ' . -
      sequence of events
      quarantine
                           285

-------
 3
 S-i
•H
 0)

 O
 O
-P
 o
      Cn
           •H
           SG
                 o
                      .
                      Cn 0)
                      •H C
                      K-H
                         fc
                       I  (0
                         3
                      •O -P
                      •H W
                      S H
 W

•H

 o


 3


 O
      cn
     •H
     as
                 I
                      ss
                       I  CO
                         3
                       5 -P
                       O W
                       i-3 W
 (0
 o
 10
•H
rH
 
-------
Ni
O
O

(0

*fl
tl
0)

y
*<






*0
,JD
O
H- ,
H-
O



rt
to t-1
0> ft>
3 3
rt
^ H-
* O
03
0) O
k^ C
F
&
cr

o
o
3
ป0
jy
rt
H-
cr
H-
rt

•



a
H-
3"







a
H-
vQ



Jpป
' ^
Q>
H-
H
(V
cr
H-
I-1
H-
rt








>3j
H-
ro
t-1
&




H-
|_i
CL
0
l_i
rj-

w
C
cr
en
rt
^
0) '
rt
,ro

50
ro.
rt
H*
o
3
i/ -
^5
ro
ro

%
h{
hj
O
ป
3*
ro
3
^
<
3
r1-
rn
H-
3
C
cr
ro
o-
ft> .
i— '
3
O
ro
i— •
H-
05
0
&)
(-•
H-
to*
t-h
           (D

^-)
pj
O
H-
Hj
H-
O





rt
t-1

rt
H-
o





a
H-
Vfi
sr







a
H-
3*






*ซป'
H-
ro
ฃ





H-
(D

p.
^
g
|_J
rt
C
i-S
ro
I-S
ro
ro

cr
c
n
n .
o
€
H-
s.
3*
D
^
t
D
-••
^
CO
3
tfi
. C
tr ,
ro

ro
" j— '
M
'H-
3
-i
b
0
rt
O
rj
[D
H-
C
W
vO
    287

-------
 Q)

 O
 O
JJ
 O
 Q)
J
          w
          V
          X
   g
   t-J
  g-
  w
-O
 <1)
4J
•H
 E
•H
J
   O
  4J
   (0

   
-------
H

ft
(D
M
 O
 O

I
 to
CO
O
3
      O
      3
W
-ft
to
ft
H-
CO
ft
H"
O
 TJ
 O

 (D
                      o
                      o
                      3
                      ft
                      M
                      O
                      I-1
                      CO


                      I
                      o
             cn
              I
             to
             U1
 +
              O
              3*

              3

              (0

              M>

              O
              3
              (D
              H>
              ffi

              (C

              O
              fl)

              c
              H-
              ft


              O1
              ro
                       •f
                              cn
                              rt
                              O
                              ^
                              H-

                              cn
               H-
               O

              ,rt
               (D
               (0
                              (D
                              O
                               [D
           289

-------
 U)
 3
 M
•H
 0}
.c
 o
 o
4J
 O
 0)
             0-
          T3  M
           10 T3
           O  C
           ^  (C
          03 CO  p..
 in
 3
•H
 ^
 O
-p
 in
 3
 (0

 O
             C-
          13  M
          f(5  >i   •
          O  (0
          ^ rH
          CQ U 0-
 (0
 O
 W
•H
rH
 (U
 o

I
             0-
          T5  W
          (0 T3
          O  C
          k  (0
          cn w P- P- 0"
 3
•H
 C

 X
 o
 G
 0}
          a   •
              u
          m  >,
          CM  (0
             r-i
          A U
           10
          4-1
          •H
          •e
       o
       o>
       &
       a>
 o

 n)

 0)
H
 O
EH
                              290

-------
Other Species


   Grandidierella japonica

   T.epidactvlus dytiscus

   CoroDhium spp
   Neanth.es ar.enaceodentata

   Meiofauna                    ,     ;   ,


other Tests      •                   •


   Microtox                      .. -     ,      ,
  - Bivalve Larval Survival/Growth/Development
   Echinoderm  Larval  Survival/Growth/Development

   Benthic Recolonization
                       291

-------
     Issues and Research•Needs

  Written Protocol - ASTM/EPA format
  Culture Protocol: Leptocheirus/Ampelisca
  Sensitivity of Cultured vs Field-collected Amphipods
* Shipping/Handling/Acclimation
  Nutrition
  Comparative Toxicology
* Reference Toxicant Control
* Reference Sediment QA/QC
  Statistical Power: Compare variability among species
  Tolerance Limits
*      Ammonia
*      Grain.Size, except Rhepoxynius
       Organic Enrichment
       Salinity, except Rhepoxynius      •
       Seasonality
       Light: Intensity/photoperiod
*  Field Validation       '
*  Interlaboratory Comparison
*  Sediment Collection, Processing, Storage
                     292

-------
              Chronic Test Methods
Test Species
      Leptocheiriis plumulosus. Ampelisca ' abdita
Response Criteria         •
      Mortality, Growth, Reproduction, Population
Dynamics
Key Issues   .
      Nutrition
   '   Narrow Tolerance Limits         .          •
'n     Chronic Control QA/QC              •   "   . ,  •    "
      Relative  Sensitivity of Acute  and  Chronic Tests
                     293

-------

-------
Bioaccumulation of Sediment-Associated Contaminants:
Significance, Current Status, and Future
Peter Landrum, NOAA Great Lakes Environmental Research Laboratory - Ann Arbor, MI


I.    Significance of Bioaccumulation

      A.     Role in toxicity assessment
             1.     Aquatic species
             2.     Human health                     .


n.    Picture of the Problem

      A.     Complexity of the exposure,environment
      B.     Current mechanistic model


El.   Criteria for Bioaccumulation Organisms

      A.     Examples of organisms

         *                -                           . •   • •              '  •
IV.   Factors Affecting Bioaccumulation
                                     -'.".,               •
      A.     External factors                                              ;
      B.     Physiological factors
             1.      Behavior
                    a.    Importance of feeding
                    b.    Feeding mechanism
                    c.    Feeding selectivity


 V.    Status                                                         ,

       A.    Field data


 VI.   Future -  Kinetics

       A.    Models                                    -
       B.    Field validation
                                        295

-------

-------
              BIOACCUMULATION

The accumulation of contaminants from all sources,
food and water.

         BIOACCUMULATION FACTOR

The ratio of the steady state concentration in the
organism resulting from multiple source accumulation
to one of the source concentrations
                                297

-------
CO
O-
••^
2L
O
h-
^
-j
Z>
^>
=>
O
O
O
CD
LL
O
UJ
O
•^
^f
O
LL
O
C/D
UJ
h-
CO
CO
CD
"D
">
O
a.

g
%— •
05
13
E
o
O
05
O
CD

CD
CO
O
a_
X
CD
ฐE
"CD ^
cง o
_co CD
M— .





c
g
j&
1
13
O
O
05
_g
-ฐ
o
CO

ฃ-
CD
Q.
CD
CD
*co
cc
"co
 o  ^
"X  13
 O  CO
•-  s
^  E
          O
          O
          O
         LL

         cJ
                              298

-------
O
                           0
                           c
o
o
-a
CD
cT
CQ
s
1
o_
^
CQ
i
















O
. r:
o"
•^
o
o
CO
co"
"01
o
^o
O '
o
i*1^
=:

•
ho

o
•
3
Z3
o

7?
(Q
~*





CD
^P
r;
Q.
^
Q.
Q.
CD"
Q.
5"
\J*
CO

0
CQ
1




"





0)
CD
O.
o"
3
CD
O
03
^^
CO*
CO
h
2_
o"

'oT
o
1— ป•
CD
3
o
"^
03
- =n
*<"
'
A
.0
.en


3
CD
/C
•— ^
— ^

Narcotic
CO
CO
CD
CD
,_+.
O
a
fS>
CO

Q.
Q.
CD
. '^^
O
X.
13"
CO






ซ— *
CD
ง
o
ฃฃ.
'oi
o
1 *^O
CT^
3
o
' ^*
ts
ro
i
o>
• MM "
3
•3

CQ
I










H
OD
O)
fn
m
O)
a
(Z
m
~n
0
zi

•H
O
X
o
H

^
:CO
0)
m
O)
0)
1 " *^
m

— i
               (Q
    299

-------
o
o
LL
O
co
z
o
LLJ
LL
O
Z
O
g
LL
H

UJ
Q
 O

LO
 D)
.E

"5

 CD
 Q_
 Q.
 13
 O)
 O
 O
 CO
 c
      CO &
 co  o
 c  E
.2  CD
 r— •+—•
-p-.  ^3
 C  O
 CD  CO
      to
      o

      0.
      o E
      E E
                O
                CO
                D)
                =L
 CO
T3  Q.
•F  Q-
^^^  "^
 03  D3
 E  a,

~o •ฃ
                  CO

               O) O
               o •
               E-!E

               T- CO
         o o
         ^^ CO
         C CD

   5    ฐ c
   W    "^ • —
               :
               ^N W
               O co
                        oS
                        LO
                        0
                           Q.
            o
            o
            Q.
                           Q
                         . c
                        c Q
                        If
                        0 —
 co ฃ
 c 2
 co "Jo
 W-R
 ^ g-
 CO CD
 c3 5-
,CO CO
         c co
         CD >,^^
         "S ^ 'O3
         e'Sf
         o S o
         2 Q. E
         O CL p
         re co fc
         C-D-
e
n
9
                        Q_ CO O
                                  300.

-------
Ingested Particles
1

\
— \
D
••?
u
D
^M*
/)
3
	 j
mi^^
[
^
•FJ
I 1
D •
•>
j
D
^^ป
u
D
•*ซ'•
•••
•)
D
0




-

























1
C
C
OM




y-n>:
s —5 s <
s CD x^
% VU x
S\NS\\\ '\\SN\X
ffSS/SSf*S/SS
Y Dissolved 1
ff-ffSSfffftSi
s UsX-
: o :;.
xO<'
\ — — s ซ
XXXV
fl
1 T
303O
5iฐ
' C X-N
-3 "
D.




























-+
'>•
JD
CD
o
C/5
~D
^"
JD
03
CD















.. -
-
Reversible
Particle Pool
1
1
TI3C
n> <*v
as
0?
o> 2
-ฐi
O CD
O






J3
co
T3
CL
*<
f
T
3 CO
O
^
*<
•
• -
m













                             o
                             0
                             03
                             0
          301

-------
MULATION ORGANISMS
•MM
Z)
o
o
<
o
m
a:
O
LL
ฃ
LU
ฃ
o



T5
CC
CD
T3
CC
"co
CO
CD
CD
&
O
•ป— *
j^
D)
CD
CD
E3
.23
25
cc
*ง
a.
i
CD
N
CO


CO
CO
co
E
	
•4— •
CO
*^
O)
"co
"5.
CL
ZJ
CO


CO
Z3
"S3
"c
CD
E
T3
CD
CO
CO
CD
.N
"•JS
E
CO
• cc
E?
o
co"

CD >>
-Q"o.
— Q.
d CO
ZJ
^cc "g
•— ฃ
1 tl
CO H~
CO
o
ollected
o
"1
o
•D
CD
ZJ
•*—>
ZJ
o
^,
CO
CD

CD
2
"5
O
De tolerant of variation in

35
ZJ
CO
CO
E
organis
•
CD
o
CO
JIJ


CO
CD
O
O
0
"c.
cc
•+— •
0
•D
C
,g
"co
o
CL
E
o
o
"c
CD
E
CD
CO

-
preferable for accumulation
CO
•4— •
c
32
"c
o
0
75
O)
i
g
1ฑ
o
o
T5
"D.

•H2
cc
c
'E
cc
•4— ป
• C
o
o
CO
o
s
CD
*4—




_Q
cc
Q.
CC
*%ป
o
g
cS
ansform
.2
JD
"cc
	 .
]c
^E
302

-------




room temperature, sensitive
sediment composition
o"








field collected, tube dweller but
laboratory conditions, lipid conter
tolerate a wide range of temperai
c
x-
CD
X
03
Q
CD
03"
CT
B
03
i
O
03
or
CD
O
C_
i— t-
CD
;=ซ• ^ -P-


i?" '

chemical concentrations





can work at room temperature, 1
sediment composition, wet weigh

^
D-
o"
C
c ,.
CO
<
03
5"
Q
03_
C
CO
CD
03
CO
ซ<"
O
C_^
f— t-
— - • 	 r~ •
-* o =^




*" -^
= '0
^ 0
CD ^
ฃ3
ง8.
3.5
SB
0 CD
0 Q.
*< Si
- S"
<
CD O~
^ 03
^ CO
CO CD
CD
CD" ^
s:ฃ
< ^r
CD V




sediment composition, size 6 m
benthos, must be field collected, t
. ,O_CQ






:
o"
o ^
O CD
if
3-TJ
^p
ro .
en '
, ^
1 CT
OlC
o 5
>,p Q.
o^ OD
0.3
-T -
*< -i
- CD
^ JD
CD C
^Q"
•— *• — — .
^^ 0
~ ฃ
o
^•^••1
er\
m
X

e*
IPLES OF FRESHWATER BIOAC
o
O 03 v-n $2- 3"
O 13  i   of   •

a ฐ- -^ CO 9:
03 O CJ1 CD =ฃ
   303 vฐ o_ ^
   ~^ o^ t^l CD
          3 C
* i
CO
JD
    3—^  fc^ซ*
    JO —

CQ  ?n  =='
CD
CO
                      CD
ฐ S

O* CD
                                                CD
                                                CD  CD
                             03
                      JD CD  =-
   S i.i  S
          Q.O.
             ^:       (Q
                          CD
                                                (O
                                                CD

                                    H,
                                    O


                                    O

                                    O
                                                          O>

                                                          CO
                         303

-------
O

<
O
O
o
CQ

O
z:


o
LJJ
LL
LL.
OD
o:
O

o
CO


ฃ


j

CD
Q.
E
CD
       CO
co    .g
0)

CO
c:
g behavio
UL     "m

CO
o
•*— ป
o
05
LL
"co
•*— ป
CD
E
c
o

c
uu
CD
ntaminant Prop
o
o


CD
diment Charact
CD
0)


^^
r-
vironmental Coi
.c
LU



to
o
o
CO
LL
"co
0
"o>
o
o
"co
_^
0.

itransformation
^
CD


v_
i5
CD
ปฃ
d)
0)
i_
o
i
CD
CD


h
ro
G
Reproductive State
                                                                    CO
                                                                    CD
                                     304

-------
Tl
CD
CD
jlD Q.
co 5-
•o 
CD <
-* o
0 Ql
CT 03
O 13
'""^ Cj
O ,
0 '
'*"*" ^

3 2-'
g. Q,
Z3 — •
03 (Q
Q. co
•-K CD
0 Q.
o 5-

•ง3
Q- _
sง
f^ ^^^
vw ^^^
55 g
*^"tป • .
S-"
^^^ •
fi^^ ~J
^ ฐ-
to
.?8
   Tl
   CD
   CD
O Q.
3. DO
CO CD

1 ^
—^ —•
CD O
Q. •"*
C  '
O rzj-
8s
8  to
    ซ
    CD
    g_

   CQ
    CO
   -O
    O
    Z5
    CO
    CD
            Tl
            O
            O
         co
            .
         0 0.
         3
         CO CD
         CD ฃ•.
         Q. O

         CD 3
         X O
          co  O
          ^•^

          1ซ
          CD-0
          03
CD

CO
CD
Q.


CD

3.

O
03
                  CO CD
                  CD X
            CD
            CD
      3-CO-
      CD  c:


     'ง>  &
      co  co

      BO
      03 CQ
      ^  CO  ?i
      8  3  i

      ^•05-
            03
         u*  m"
         —_. {\j
      en'3  ฐ-
      ^  CO  _^

      co  2. ง
            Q_
a
ami
O  —. Q
3  5-ป
— 03
CQ -3 3
                   CO 03
                         CO
                   ป-+• -3 —
                   S'
                      O CD
            DO
            m
                               <
                               O
      =r    o
            O
            m
            C/D
            >
            Tl
            Tl
            m
            o
                               O
                               CD
                               o
                               o
             I
             o
305

-------
O
z;
Q
LU
LLJ
LL
LL
O
UJ
O


I
CL
O
CL
          O)
CD
CD
                          c\j
+1
in
CD
C\J
      CM

      +1

      CD
      CO
      CD
00
CO

C\f

+1


CD
CM
                            O
                            CM
c\
+1
CO
         LO   O>    CO    CO




o>^
c -^
~5 ^E
CD CL
CD o
ii j^i.
CO
CO
^M.
CM
+1
00
CD
^t
LT>

0)
CO
CM
co"
+1
O)
00
O)
c\f

00
o
CD^
crT
HH
00

T—
T-T
CM
^
T^-
C^
CD"
+1
T_
CM
CM
oT
CM
          o
          "
JZ


CM
                          CM
                      O
                      CM
                   00
                   CD
                                  306

-------
  Accumulation Factors
       For PCB Congeners
 52 101 151 118 153 138 128 180 195 194

            lUPACNo.
N. incisa
Glycera
M. mercenana
               307

-------
 0)
 CD
CC


 CO

 U)
 o

CL

CC

               O
/•!••:
vL
                E
                O)
                Q
                *

               rT
C/D     'ฑ:  ^
        x" eg
        0 CS
       T3  SP'CD
       • ฃ -c  -i

       "^  w  cu
        ^  ง  "E
        x  S^  o
       LU  C  o
                    Q
                    cu
 0  0
 O  0
 CC *""""
J=  0


ll  ^
"SS- fp -^
 Cu  ^  o^
"T  0  '*ป'

1;'ฐฐ  S.

Q *ฃ. ฃ
                                308

-------


^
o
c:
CD
-13
i— h
3
CD
.ป— h.
ZT
O
Q.
O5
O
03
0
I3_
^
o
o
o
r-*-
6*
— t
Q3
>**
C/5
-j"
CQ
CD
C/5
O
O
CD


'


GO
O
13"
CQ
CD
05
o
CQ
03
o3"
3
O5
o'
0
CQ
^
=•
O
^^
03
13
CQ
CD
CD
O5
c_
oT






ro
0
03_
03
O5"
O
O
g.
6"
o
.3
CD
X
CD
1*
CD
13
•— ^
05
O
o
1 M^
Q.
r-+-"
o"
13
C/5









•7*
O
c .
CD
Z5
•— *•
3
CD
5
o
Q.
05
CD
' "D
O
i— t-
CD
O5^
I"
CD
O5












D
o
m
O
c
33
m
"Z.
^
m
T
IODOLO
Q
"U
O

6
m

"Z.
m
m
D
m
D

•T|
O
33
H
O
309

-------
            CO



f^_
o-
CO
O
h-
UJ
-z.
^
X
^











S2
ง
to
^
CD
-t— ป
CO
c
05
•*— •
CD
"o
•*— ป
O5

0
5
1
=3
O
O
05
t •
.2
CD
ol
•*— ป
c
o
E
"*— - r"
D)
_O
CO
CD
^M,
"5
CD
E
CO
"c
05
D)
L_
O


^ •+—'
, +z to
i-^
>s^
O5 to
to >
^ CD
*•!
E co
a.
ss of contamina
_0
T3
05
0
78
. 13
E
13
0
O
05
D)
"o
CD
ซT
CO
E
"c
05
O
CD
E
CD

05
X
UJ
E
CO
Wrf
o
Q.
X
CD
o
CO
^1^
^^J
o
_CD
•"5
E
E
:B
**~
c .
"•g
"5
E
13
O
O
05
 ^a^
*^ป ^uv
•— J
CD E
C3) Q.
ฃZ n^
Z? vM
O5 ป^-
o xf
ft
^^ CJ)
^3 ปL
0 D)
g CD
<;


o
05
O
cL
a.
O5
CD
Z3
T3
"co
E
CD
^3
CO
CO
"•*=
CD
•i--
D)
"co
13
+2
1
. .
0
T3
E
a.
c\
CO
in
CJD
         310

-------
      "D
03
O
CD
CD
(O
CD
ป— K
o"
cn
03
03
C/)
CD
Q.

~D
^^>ป
C/5
6"
o
(Q
o"
.SL-
03
C/5
CD
Q.
ฃ2.
o
o
(Q
o"

CD
C/)
CD
Q.



O
CD
Q3
03
^^
O
CD
O
' C
CD



• T1
CO
03
0
ปF->
^





                           Zl
O
o
3
                           CD     ^>
                           O
                           o
                           o
                           CD"
                           '13
CD
OD
03
05
CD
Q.
                                             O
                                             a
                                             m
                                             03
03
O
O
CD
r—t-

O
o
Q.
CD_
O>
              311

-------
E!
o 2
S— .=
           312

-------
00
                                313

-------

-------
Bioaccumulation of Sediment-Associated Contaminants:
Present Status, Laboratory Methods,  and Related Research Needs
Henry Lee II, U.S. EPA Environmental Research Laboratory - Pacific Division
I.    Important of Bioaccumulation of Sediment-Associated Compounds


n.    Methods to Measure/Predict Bioaccumulation of Sediment-Associated Compounds

      A.     Criteria to choose among the methods


HI.   Equilibrium Participating Bioaccumulation Model

      A.     Use as screening tool
      B.     Limitations/uncertainties with equilibrium predictions
      C.     Lab/kinetic alternatives to partitioning paradigm


IV.   28-Day Bedded Sediment Bioaccumulation Test

      A.     Status of "standard 28-day" sediment bioaccumulation test
      B.     Test duration                             ,
        .  "  1.     Why need a 28-day test vs. 10-day test
             2.     Adequacy of 28-day tests for slowly accumulated compounds
      C.     Organism selection
             1.     Selection criteria     .'.,",
             2.     Why need sediment ingesting organism
             3.     Recommended bioaccumulation species
      D.     Laboratory methods
             1."   No feeding of test organisms
      E.     Experimental design
             1.     Number of replicates and statistical power
             2.     Pseudoreplication           >
      F.     How proposed methods differ from those in the "green book"


 V.    Toxicokinetic Bioaccumulation Models/Tests

       A.    Equilibrium vs. non-equilibrium exposures
  •'•.   B.     Modifications of laboratory procedures                         .
                                        315

-------
VI.    Evaluation of Bioaccumulation Results

       A.   . Criteria .for "reference" sites


VH.   Research Needs

       A.    Related to 28-day bioaccumulation test
       B.    Field validation
       C.    Round robin
       D.    Lipid methods
       E.    Research needs related to other methods of predicting bioaccumulation
       F.    Resuspended sediment tests
                                        316

-------
317

-------
  PREDICTING BIO ACCUMULATION OF SEDIMENT -
ASSOCIATED POLLUTANTS BY INFAUNAL ORGANISMS

  Field Approach:
       measure  tissue   residues   in  species  collected
       at surrogate site

  Bioaccumulation Test:
       measure   tissue   residues   in   species   exposed  to
       sediment collected at a surrogate site

  Bioaccumulation Factors (BAF):
       ratio of concentrations in tissue to sediment         .

  Thermodynamic Partitioning Model:
       based   on   chemical   partitioning   normalizing
       for  lipids   and   TOC   but  only   for   neutral
       organics

  1st Order Kinetic Model:
       tissue   residues   modeled   as   balance   between
       rate of uptake and rate of loss

  Toxicokinetic Model:
       tissue  residues   modeled  as   function  of
       bioenergetics  of the organism
                        ' 318

-------
Figure 2.1: Summary of questionnaire choices. Numbers above decision boxes refer to
questionnaire choices.
I'.'''1" ' '"



Predict or model tissue residues

1 Use EqPB Model 1
Section 4 1

Compound is
polar organic?
12
I UseBAFmc
1 Section 3
' • •"•"••""^
Use a standard model to
residues.

s metal or Yes

__^__ Go to Next Page for Field or Lab •
questions (15) • ' ' , '
Yes . " '
predict


Model used as quick screening tool
for bioaccumulation? •
Yes "' ' . 5'

xiel, 1
Model more man one u]
(as opposed to modeling
from bedded sediment?
6
No
stake route
uptake only
1 Collect field s:
regression moc
implcs to derive a 1 •
lei. Section 11. 1
. .
Iljsc bioenerge
toxicokinetic u
No 7
Will sediment pollute cone, change?
8
No
Predict time to specific tissue
residue value (time to steady-state
or elimination time)?
Kre site-speeificYAAFs
/

11

No
Are specific k2 and ks Yes
values available?

'Yes
9
No
Will organism grow substantially
during test or modeling?
10
Yes Changes in test
are likely due t
characteristics.


Yes 1 Use Ist-order
1 Section 6
No
Is compound metal or polar
organic?


,'..
14
Is measured AF available for
compound and sediment of concern?
1 Use Ist-order kinetic 1 '
model 1
Section 6 |

ics-based ' 1
nodel. Section 5. 1
'Yes '-..-.'
organism behavior
o changes in sediment
No .
kinetic model 1
Yes
No Specific ks and k2 values are both
available
Yes .
[Use EqPB Model 1 •
Section 4 |
Accurate est
(box 4) or n

No
imate not possible. 1
screening prediction 1
lake site-specific 1
t fRnr 1 SI I
319

-------
I *
ง
                                V
ns
ปPN
ft
.5
• PN

05-
         A  1
         T  4-
            fi
            o
?• -8
JU  H
U
O
H
^ tซ i-j
y, ฐ^
               to
               U
            II
                   U
            cc
                         bฃ  S
                      y\
                      DD

                      "Sb
                      s
                      ^
                      0>
                      ซM
                      C8
                      S
                      * *•*
                             a  ^
                             g  ^
                             S  a
                             5
• fl   *


 S^'T
pg ^o ง
 S M) tj
                 2 -^ CQ S • *—'
                 urn
                        ,.
                        "
                         U
                       8
                      JS  O w.fa
                      U J HU<
by partition
                                   1
                                      t
55
ฃ;
S
o
|
*>
^S
•s
>J
tf-
K
^ป
5
1
ซa
. *?

•**

                                   •i
               320

-------
. Table 4.1 Continued ,

COMPOUND
HCBP
HCBP
HCBP •
HCBP
HCBP
, HCBP
HCBP
HCBP
HCBP
HCBP
HCBP
HCBP
HCBP
HCBP
HCBP
'HCBP
HCBP
HCBP
HCBP
HCBP
•'' , HCBP
HCBP
HCBP
HCBP '
HCBP



PESTICIDES
a-chlor
a-chlor
, ; g-chlor
g-chlor

DDD
DDD
DDD
DDD
DDD
DDD
DDD
DDD
DDD
DDD



DDE
DDE
DDE
DDE
DDE
1 DDE
- - DDE '
DDE
DDE




ORGANISM
Yoldia limarula
Yoldia limatuia
Yoldia limatuia
Yoldia limatuia •
Nephrys incisa
Nephtys incisa
Nephrys incisa
Nephrys incisa
Nephrys incisa
Nephrys incisa
Gtycera sp.
Mercenaria mercenaria
Nereis virens
Nereis virens
Nereis virens
Nereis virens
Macoma nasuta
Macoma nasuta
Macoma nasuta
Macoma nasuta
Macoma nasuta
Macoma nasuta
Diporeia sp.
Oligochaetes
Palaemonetes pugio
Bivalve Mean
Polychaete Mean
' OveraU Mean

Yoldia limatuia
Nephrys incisa
Yoldia litnatula
Nephtys incisa
OveraU Mean
Macoma nasuta
Macoma nasuta
Macoma nasuta
Macoma nasuta ' .
Macoma nasuta
Macoma nasuta
Yoldia limatuia • ^"
Yoldia limatuia
Nephrys incisa
Nephtys incisa
Bivalve Mean
Polychaete Mean
Overall Mean
Macoma nasuta
Macoma nasuta
Macoma nasuta
Macoma nasuta
Macoma nasuta
Macoma nasuta
Oligochaetes
Oligochaetes
Oligochaetes
Bivalve Mean
Overall Mean
• •'

AF
1.7
0.9
S.26
5.1'
0.4-
0.2
16.6'
7.1'
7.1'
3.81
3.9
1.5
4.8.
1.4 ',
4.8
1.4
4.8
0.5
1.6
2.7
, 4.0
1.8
0.7s
1.2"
2.1
• 4.4
4.7; '
4.4

4.0
4.2
4.5
5.9
4.7
1.0
0.4
0.7
0.7
0.5
0.5
4.0 ,
4.2
4.2
4.8
1.5
4.5
,2.1
2.8 •
0.7
1.3
1-1
0.7.
1.1
0.9'ฐ
1.0'ฐ
1.3™
1.3
1.2


TOC% .
1.9
4.0


1.9
4.0






1.1 '


5.7-
1.1

0.8 ,
1.3
2.5
5.7
•0.7
" 3.6
5.7









0.9 .
0.8
3.7
4.0
5-l
7.4







0.9
0.8
3.7
4.0
5.1
7.4
3.6
4.6
6.0


321

LIPID%















7.6


4.6
4.6
4.6
1.3

6.6
1.6









5.5
5.5
5,5
5.5
5.5
5.5







5.5
5.5
5.5
5.5
5.5
5.5
6.6
6.6
6.6



• • ' .i
REFERENCE ,
McElroy and Means, 1988 ' :
McElroy and Means,' 1988 . .
Lake et al.. 1990
Lakeetal.. 1990
McElroy and Means. 19S8 '
McElroy and Means. 1988
Lake et al.. 1990
Lake et al., 1990
Lakeetal.. 1990 .
Lake et al.. 1990
Lake et al:. 1990
Lake et al., 1990
' Brannonet al., (manuscript) '
Brannonetal., (manuscript)
Brannon et al.. (manuscript)
Pruell et al.. (manuscript)
Pruell et al., (manuscript)
Pruell et al., (manuscript)
Boese et al.. (manuscript)
Boese et al., (manuscript)
Boese et al., (manuscript)
Pruell et al., (manuscript)
Landrum (pers. comzn.) .
. Olivier, 1984
Pruell et al., (manuscript)- • <




• Lake et al.. 1987 ' '. ,'
. Lakeetal.. 1987 '
Lakeetal.. 1987
Lake et al., 1987

Ferraro et al., 1990
Ferraroetal., 1990
Ferraroetal.. 1990
Ferraroetal., 1990
Ferraro et al.. 1990
Ferraro et al.. 1990 ,
Lake et al.. 1987
Lake et al., 1987 ' .
Lakeetal., 1987
Lakeetal., 1987



Ferraro et al., 1990
Ferraro et al., 1990
Ferraro et al., 1990
Ferraroetal.. 1990
Ferraro et al., 1990 '
Ferraro et al.. 1990
Olivier. 1984
Olivier, 1984
Olivier, 1984


t

-------
~m—*   •—-


A"!
 0)
    06
O  ซ

*  a
    s
                          322

-------
                                       — •  y
          CO
          CD
          co
          CD
         GE-
          CD
         -D
          to
          CO
              Idealized Uptake. - Elimination  Curve

              '  Cf(f).= Cs *  Ks/|<2  •  (  l-e~k2' ')

               Uptake Phase              'Elimination Phase
               .•-Cs =  k 2* CI ;

                 Sleody-slale  \  Fosf component
         Linear uptake phase
                         Cs ป k.
                      CI
                                  Slow component
                                          I
                  (Contaminated Sediment)  \ (Clean Sediment)
Ci = tissue—concentration (ug/g)
Ks = sediment uptake rote  coetd'cienl  (
Cs = sediment concentration (ug/g)

k2 = elimination constant (I/time)
f  = time
                                                     edimcn>   .

                                                         * ''mฐ
                 Figure, 2.   Idealized uptake — elimination curve.
            BIOENERGET1C-BASED  TOXICOKINET1C MODEL
                 MAJOR UPTAKE  ROUTES FOR DEPOSIT-FEEDERS
                                  Sorplion/Dermol  Uptoke

        Tissue Residue  —  Ow  + fw 4-  Si .  •*• Ss + SprpJiori — Eliminalion
                      /   Flux
Uptake Sediment  = t Sedimenl
                            \   ( Pbllulanl
                            ; ' V   Cone.
                                                        .V   ( ExIrocJion  \
                                                        / "\ CffJciency ./
                               /
	 Flux Sgdtment  = /fWgtqhl. Activiiy. TQCJ
         Ss — Susp-ended Solids Up-lako Roule
         Qw =  Ovgrlyin-q  Wgter Up-ltrtte Rrrule
         Si = •lh'g*S'Jซ''d Se-dim-e-rH Upซcrtce R:ovte
         (w =
                                                                           _J
                                323

-------

fฃ> -
fl)
imr
Z5
"O

	 ^i^. . ,, •
CD
. .Qi a
H— •
ni ^
2 w
-3 D
I// .X""
c/i
*~ E
"D ฐ
0 .0
> ,9?
CD
C/) ฃ-
JD *""
X1— N j_^
— o
CO ^"
> m
cป^
(^ O
0)

'
T3
CD
L.
CL
•
•















_Q
Q.
Q.

o
c
o

c
CD
a>
c
o
o








'.. . • • • / ' . '




.;

- • ' - . • .

* 'X
•


.'•'''



*^





"" .
1 1 II
o o o o c
o o o o
o in o in
CM ^— t—





o>
^~ • .


CM
O)

CO
e-\ >*ป
-g ฃ
CD
ft^N *••
<0 .K-
0)
3
CO
CM O
*


324

-------
                                          03
                                          O
                                          X
                                                      X
                                                      CD
                                                      i
                                                      o
                                                      ง

                                                      3D
                                                      O
                                                      n

                                                      I
                                                      CD
                                                      •o
                                                      •I
325
                                                      c
                                                      CD

-------
 EVALUATION OF DREDGE MATERIAL BEDDED -
      SEDIMENT BIOACCUMULATION TEST

OBJECTIVE:  Measure   tissue   residues  in  infaunal
organisms   resulting   from   exposure • to   dredge
material

PREVIOUS  APPROACH:   10-day  bedded-sediment
bioaccumulation  test  to   estimate
"bioaccumulation potential"
                                            "i
PRESENT   APPROACH:   28-day   bedded-sediment
bioaccumulation  test  to   estimate  "steady-state"
tissue  residues   (10   day   -  test   if  only  metals
present).  Used in Tier HI

STATUS:   EPA   Guidance   Document  produced   and
referenced   in   Implementation   Manual.     Guidance
document in ASTM review
                      326

-------
O

C/2
s
MJ
z
w
o
3
Cd
W
13
ง
H
3
n
H
^^
[VE OF THE MAJORITY
O:
T]

.00
13
8-
i~<
W
oo


















G
z
r
w
00
00
E
B
00 •
G
r
11
ARE SPECIFIC TO
> .

oo
HH
ง '
5
oo
a-
n
S
\ป

H
Cd

H
oo ,
H

oo
8
O
G
r
0


td
W
d o
55 J#
oo Q
G :>•
MSMS, ALL T]
E RESIDUES.
w
W
REASONS REQUIRE
>
h_^
•g
ASONABLE

M
oo
3
|
•'. 3

o


oo
W
J

i
oo
[~j
^

W
W
X
o
tfl
I
a
N
|
IDENTIFICATION /
0 .


EXPOSURE

H "'
_

1
W^
7
r .
<
tn . .
D

td
W
ง
H
a

n
0



SCIENTIFIC UNDE
90
00
H
ENDING OF SEDIMENT
s
o

^
rAILABILIT^
•^><
O'
-
o
oo
..6 :













0



TEST OR DERIVE
1
h^
MENT QUALITY CRITE
.2





















0



ASSESS EXPOSUR]
CO
H
O
DEMERSAL FISHES, M
1

w "
r
>*
>
Q
s

*"












0 ~~



ASSESS SEDIMEN1
^T
^^
DR HUMAN HEALTH EF
1
\ j
H



















0' • ,



ASSESS EXPOSUR]
en
3
BENTHIC ORGANISMS
s
xw
r ^
m
iCOLOGICAl

. • s .
oo
W
oo
1
oo

I
H
oo








0



ASSESS DREDGE 1
".ง
gj
oo






















0 •



HAZARD IDENTIF
r-*
i
ION (BIOACCUMULATII
o

0
I
1






















."a.
o
3
0
d
CT SEDIMENT
w
^,
^^^
o
^
Jj2
r

H
0
' ^^
•^
H

cc
H
• -^
^r





327

-------
                                    ERLN-N111
r
\
     UssyEPA
     GUIDANCE MANUAL:
     BEDDED SEDIMENT
     BIQACCUMIILAHOJNL TESTS
     U:S. EnvironmentarPr otec tioh "Agency
     ERL-N
     Pacific Ecosystems Branch
     Bioaccumulatlon Team
     Newport, OR/

                   328

-------
      Sediment Bioaccumulation Test
            Key Procedures
1.     28-DAY EXPOSURE DURATION.
                      '      *      '   •
2.     SEDIMENT-INGESTING   ORGANISM
      REQUIRED.

3.     NO SUPPLEMENTAL FOOD USED.

4.     S P E  C I E S    EXPOS  ED
      INDEPENDENTLY.

5.     80%   OF   STEADY-STATE  TISSUE
      RESIDUES   RECOMMENDED
      ACCURACY.

6.     L Q.N.-G -TERM  T E S T S   OR
      TOXICOKINETIC   APPROACHES   USED
      FOR  > 80% ACCURACY  OR  SLOWLY
      ACCUMULATED COMPOUNDS.
                    329

-------
330

-------
oo
CO
d
HH
H
W
S
Tl
0
^ป
MECHA
•2:
HH
&3.
1— H
O
ฃ
z •
^-— • <
3
h™^
O
00
H
S
ฃ^
w
00

.tl
W
W
a
^
o
w
tn
ffi
^^
HH
0
d..
0
w

o
o
0








K-
I-H
O
ffi
W
H- (
0
o
0
rl
fMULATl
0
.2!
O
. w
H
1— H
r







CO
d
HH
Q
W
H
dd

o
00
00
0
O
ffi
w
ง
n
: r
^
^
>
r
^
00
^^^
^^^
oo
>
00
t— 1
r

n
0
.•F
w
o
H
w-
a
o
o
d
d
a
.- -







                               HH

                               HM
                                      H


                                      d
                                      1J

                                      W
                                      •2;
                                      o
COLOUS
CO

a

S
w.

H
O
W
oo

3
*J
      O

      S
      H
      H.
      ^
      '3.
                                                   r ~

                                                   O
                                                   c^
                                                   H
                                                   S
                                                   n
                                                   H

                                                   I
                         331

-------








to
C
y
M
to
CO
H
I
O
CO
U
[~<
1
U
2
1
b
'ERTINEN1
(s<












.2|
*"
.1 -&
S =
i —
SCOT
5
0 >
2 ."2
"a -2
U ฎ
w p^

Pollution
Tolerance


_s
02
ซ
R
.S

ex
.ฃ *












H
+ + i



' + +



. , ,



+ + ,


>n ปn o
A A A


-i- + +

i ง S
CL< to wa
" ** CO












Abarenicola spp.
Arenicola spp.
Callianassa spp.
+ + + + +'"+'+ + + + '
+ H- + + + + +


+'••+++ i +" •


, ' ' ' J
+ '• • • t . • i i + • g.
"S ' c <-> '.
ง -

ooovnmoo oซn
AAAAAAA^'AA
"S
^j ^5 •
~r" C*~* ™r~ ~t~ C^** ^^ ^5 ^^
' "T" ~i — 1— I, i J_ ^^ *^ ^D
. fS u .-f
= o"ง 1 '
^fe^QfeQQQfcงfcOOH ^
@@eง@ggงg-ง . |
1 - 1
s ฐ
_ S^ cC

•O „_. ti S
eu .ti o _
CO so *Q C!
rt to ฎ l> O
I . '. ' 'I |"^ .1

o _ ' 8 u <3 •ง •
*2 fcซ O eu o ^- ' S
% - S 5 "g>- O eg to |
. -J 1 - 8 . 5 - & S -8 1 -a 1
eL * S .2 S ซซ S ซs • P .s .S s >-H
S3 i J B 1 ง 8:1 1 P| ii
If i BlTllll Tjf 
-------
o
ฃ
Si
                          '*
                                       11
                                       II
                                       S.
                                        •f
                                             4
                                             ซ-• ST.
                                             a- S
                                               I
*o
S
o
              a
               t
               o


               o.
          rt>
          o

          on
"O
o
o
                          T3 O.  co,

                           i.1  i
                             3- 
-------
                                        (ฃ>
"D
 (D
 2
     CD
 N
 C
 O
 ^  c
    *""
D  eo
X  D
CD  C

     O
 -  ฃ
o  o
     o
CO  D
0
0
D
a.
                                                                    •o
                                                                    a>
                                                                    Q)
                                                                    D>
                                                                    ฃ
                                   334

-------
p_
s\
^4*.
? _
H
"•ป
P
o
CD
CD
rf-
P_
-
i—
IS
>o























II
P
OO

(TO
•
O
•Pi
^
" S-' ' '
sr
to

5s
,2-
•pi

-~ป.
2
p
ฃ '
(TQ
#
O
, -pi .-.











w hd >^
gn CD O
CD cซ J^J
E,:C?- S?-
ff. , .- - - ^^ _
^ a
CD





o - w . o
^ Cri O







O H- <|
Ni ^ tO
•— OS
00





to i— bo
tTl "^ ^^
^ to
OO \O,


^^ I— ' , -ปJ
to ^ oo

1— tO H-
OO J^ ^^









Q CD E^ -^
G G. CD . ^
v-, •"* ' ง

2.
P


*- ^TJ o iz;
^~^ >P CD CD
t^ o ^^ CD
P w ' w 55*
-03 ' *~^




tO ITJ M 21
OO CD CD CD
Ho. "CD
^ 2ป 2. So"

CB S"' "
to Tj M :g
00 CD CD g1
nil
O5 P
III
O O
11.^













x

d
S*
S9
1 ff
a o.
85 -. *
5s
is> H 5'
C/5 *^
ei *0
jd "^

-------
is:
ad.
                                                   JO
                                              336

-------
r=r=r~(%f& WT.}
'(Thousands)

-------



o
o
II
fc

*O
0
U


CO
H
CO
>

Reference
test.
o
?-i
I-H
"S
W
CO .
/I \
1
0
CO
CO
s

O
IcL
• 1
wi
o
a

-------
ZD
ซ
—.14
3**
ฃ *
^ e
4
3
A
Sediment
I -

-
b . ซ

Tta ~ -" nrtMMiB
• ^

k

Rซf*r*noซ
8*dlm*nt

to



^








is
i*
i*
4
a
A
TซSt
Sediment
r
^
- ^
^ ," -ซ
r
H
8*dlmซnt

s
•
N
H

h

                                                             10
                    NfertabMy
             Un
C
 ipatabli
         Test
         aedime
                               10
              I    I    I
                                                   MMU
 Appendix Figure  1:  Acceptable and Unacceptable Reference
 Sediments.  (A) Acceptable  reference  sediment with test confidence
 intervals not bracketing the  tissue  criterion.   (B)  Acceptable
 with test confidence  intervals bracketing the tissue criterion.
 (C) Unacceptable because reference confidence intervals overlap
 the confidence intervals of a mean test residue exceeding the
 criterion.   CD)  Unacceptable  because reference confidence
 intervals exceed the  tissue criterion (test confidence intervals
—ฑnBnarerฑarrr~~Th'e  points "represenfth'e tissue residues "that "would
 result  in organisms exposed to a particular sediment
 concentration.   A BAF of 2 was used  for illustrative purposes
 only.
                                 339

-------
                                       vs..
       "Guidance Manual" and "Synthesis of Methods"

                                   GUIDANCE/SYNTHESIS DOCUMENTS
DREDGE MANUAL

EqP BIOACCUMULATION MODEL SCREEN
FOR NEUTRAL ORGANICS

"THEORETICAL BIOACCUMULATION
POTENTIAL"
GOAL OF TEST =  BIOACCUMULATION
POTENTIAL"

10 DAYS FOR METALS
28 DAYS FOR ORGANICS

N = 5
                                  . EqP SCREEN FOR NEUTRAL ORGANICS
                                   BAF'S SCREEN FOR METALS/POLAR
EQUIVALENT TO USING EqP
BIOACCUMULATION MODEL WITH AF = 4

GOAL OF TEST = TISSUE RESIDUES
> = 80% OF STEADY STATE RESIDUES

28  DAYS  FOR ALL COMPOUNDS  AS
"STANDARD" SINGLE POINT ESTIMATE

REPLICATION (n~ =8) BASED ON:
TYPE I ERROR = TYPE II ERROR
DETECT 2-FOLD DIFFERENCE
INCLUDES FILTER-FEEDING ORGANISMS    ONLY SEDIMENT-INGESTING ORGANISMS
TO CONTROL SAMPLES  ANALYZED  IF
"DISCREPANCIES"

ALWAYS PURGE FOR 24 HOURS '
CONDUCT TIER IV  KINETIC OR FIELD
EVALUATION ONLY IF FAIL TIER III
KINETIC TEST OR FIELD EVALUATION IF
FAIL TIER IE
KINETIC  APPROACH WITH  kl  AND  k2
ESTIMATED FROM UPTAKE CURVE ONLY

NO  QUANTITATIVE  CRITERIA  FOR
CONTROL SEDIMENTS

NO  QUANTITATIVE  CRITERIA  FOR
REFERENCE SITES
                                  TO  CONTROL  SAMPLES  ALWAYS
                                  ANALYZED

                                  PURGE FOR  24  HOURS  EXCEPT FOR
                                  TROPHIC  TRANSPORT  STUDIES  AND
                                  RAPIDLY METABOLIZED COMPOUNDS

                                  KINETIC,  LONG-TERM EXPOSURES, OR
                                  FIELD  STUDY  DEPEND  ON  GOALS,
                                  COMPOUNDS,  RESOURCES, AND
                                  ACCURACY

                                  (FOCUS ON EFFECTS OF TISSUE RESIDUES
                                  RATHER MORE ACCURATELY ESTIMATING
                                  TISSUE RESIDUES)

                                  KINETIC APPROACH WITH kS (=kl) AND k2
                                  ESTIMATED INDEPENDENTLY

                                  GUIDELINES FOR ACCEPTABLE SEDIMENT
                                  POLLUTANT CONC. IN CONTROLS

                                  STATISTICAL  CRITERIA   BASED  ON
                                  OVERLAP WITH  TISSUE  RESIDUE
                                  "CRITERIA"
                                  340

-------

-------
342

-------
          BEDDED SEDIMENT BIOACCUMULATION TEST
                     RESEARCH NEEDS
I.   INTERLABORATORY ROUND ROBIN

H.   FIELD VALIDATION

IE.  ADEQUACY OF 28 DAY TESTS

IV.  REFINEMENT  OF  CULTURING  AND  EXPOSURE METHODS  FOR
    STANDARDIZED SPECIES
         1.    BIOLOGICAL: TEMPERATURE, SALINITY, GRAIN SIZE,
              TOC
         2.    EXPOSURE: SEDIMENT/ORGANISM MASS, GUT PURGING

V.   EFFECTS OF SEDIMENT COLLECTION, STORAGE, AND SPIKING ON
    BIOAVAILABILITY

VI.   "STANDARDIZATION" OF LIPID METHOD

VH. TESTING/GUIDANCE FOR NEED FOR 2 BIOACCUMULATION TEST SPECIES
                                          '
VIE.      DEVELOPMENT OF REFERENCE BIOACCUMULATION SEDIMENT

K.   REFINEMENT OF STATISTICAL DESIGNS
                •••/.•     •    •      '     '     -      .
X.   DEVELOPMENT /VALID ATIONOF KINETIC ALTERNATIVES TO 28-DAY
     TEST

XI.   TEST SPECIES FOR SUBTROPICAL SUBARCTIC, AND OLIGOHALINE
     HABITATS          ,               .
                              343

-------
        BIOAVMLABELITY OF SEDIMENT CONTAMINANTS
                     RESEARCH NEEDS
I.   QUANTIFY  UNCERTAINTY/ASSUMPTIONS  OF  EQUILIBRIUM
    PARTITIONING BIO ACCUMULATION MODEL

H.   DEVELOP SCREENING MODEL/APPROACH FOR METALS

m.  DEVELOP METHODS TO  QUANTIFY  EXPOSURE,  INCLUDING
    INGESTED DOSE, OF BENTHIC SPECIES

IV.  DEVELOP/VALIDATE TOXICOKINETIC APPROACHES FOR BENTHIC
    SPECIES

V.   COUPLE TOXICOKINETIC AND TOXICODYNAMIC APPROACHES TO
   • PREDICT TISSUE RESIDUE EFFECTS ON BENTHOS

VI.  PREDICT TISSUE RESIDUE EFFECTS FROM "WEIGHT OF EVIDENCE"
    OR USE AS INDEPENDENT VARIABLE IN AET'S
                             344

-------
(log) RISK

-------

-------
 Discussion of the Use of Lumbriculus variegatus
 in Freshwater Sediments
 Gary Ankley,  U.S. EPA Environmental Research Laboratory - Duluth, MN
 I.     Desirable Attributes in Selection of Species for Sediment Bioaccumulation Testing

       A.     Readily available
       B.     Known exposure history
       C.     Adequate tissue mass for trace analyses
       D.     Easily handled
;       E.     Amenable to long-term exposures    -,.-••"••               .
       F.     Reflect concentrations of contaminants in  field  organisms  (i.e., exposure is
              realistic)
       G.     Tolerant of a wide range of physico-chemical conditions in sediments (e.g.,
              particle size)             ,
 n.    Freshwater Species Uses for Sediment Bioaccumulation Testing

       A.    Chironomids
       B.    Amphipods        •••".,                            .''.••
       C.    Mayflies
       D.    Clams
       E.    Fishes
       F.    Oligochaetes"            .                             .

 ffl.   Oligochaetes as Bioaccumulation Test Species

       A.    Certain  species easily cultured and, therefore, readily available with known
              exposure history
       B.    Provide adequate tissue mass for trace residue analysis
       C.    Can be used in long-term exposures
       D.    Easily handled and tolerant of a wide range of physico-chemical conditions
       E.    Realistic exposure to sediment-associated contaminants

 TV.   Attributes of Lumbriculus variegatus

       A.    Relatively large (-5-10 mg/organism)             V
       B.    Easily .handled    ',.                                     ,
       C.    Tolerant of wide range of physico-chemical conditions (e.g., DO, particle size)
       D.    Tolerant of many contaminants-
       E.    Can be used in long-term tests
       F.    Standard culture and test methods have been developed  /
       G.    Some field validation.                           •'>..-.
                                          347

-------
V.     Lumbriculus variegatus in Aquatic Toxicological Studies

       A.    Single chemical toxicity testing  (Bailey and Liu, 1980;  Ewell et al., 1986;
             Nebekeretal., 1989)
       B.    Sediment toxicity testing (Ankley et al., 1991a; 1991b; Gall et al., 1991; Carlson
             et al., 1991; Phipps et al., 1992; West et al., 1993; Dermott and Munawar, 1993)
       C.    Metal bioaccumulation studies (Ankley et al., 199Ib; 1993; Carlson et al., 1991)
       D.    Nonionic organic bioaccumulation studies (Ankley et al., 1992; Call et al., 1991;
             Nebeker et al., 1989; Schuytema et al., 1990)
                                        348

-------
,,  Identification of Long Term Needs for Assessing Sediments
  Norm Rubinstein, U.S. EPA Environmental Research Laboratory - Narragansett, RI
  I.     Major Goals

         A.    Identify "problem" sediments
                                                   -      •  f
         B.    Assess potential impact of contaminated sediments on aquatic habitats,
               wildlife, and human health

         C.    Remediate  contaminated  sediment  sites  in  a  cost-effective  and
               environmentally consistent manner                        :.'.-'•


  n.     Scientific Questions and Research Needs

         A.    What are the  most technically valid and  cost-effective approaches for
               deriving sediment quality criteria?

         B.    How can we best identify and quantify contaminated sediment exposure
               regimes?

         C.    What are the key physico-chemical factors controlling the biological
               availability of sediment-^associated contaminants?

         D.    What is the ecological and human health significance of sediment mediated
               tissue residues in aquatic food chains?

         E.    What kinds of assessment methods are needed to best identify ecologically
               relevant endpoints and how can these techniques fit within a tiered testing '
               strategy for eco-risk assessment?

         F.    Which specific fractions or individual constituents of sediment-associated
               pollutants are of significant lexicological concern?

         G.    What are the best approaches for identifying and cleaning up contaminated
               sediment sites?

         H.    To what extent is natural recovery sufficient to remediate contaminated
               sites?
                                           349

-------
       Research Areas

       A.    Sediment quality criteria

       B.    Contaminated sediment assessment methods
                                                             *•     '
       C.    Remediation technology

       D.    Monitoring


IV.    Sediment Quality Criteria (SQC)

       A.    Field validation of EqP approach

       B.    SQC  development for ionizable  organic  and metallics (e.g.,  AVS
             approach)

       C.    Development of  SQC tissue residue approach to  address wildlife and
             human health concerns


V.     Exposure Assessment

       A.    Exposure assessment modeling for aquatic disposal  of dredged materials

       B.    Wasteload allocation  models to evaluate contaminated sediments and
             source control options

       C. .   Chemical analytical methods development


VI.    Effects Assessment

       A.    Development and validation of acute and chronic  testing protocols for
             contaminated sediments in freshwater and marine systems

       B.     Development and validation of contaminated sediment bioaccumulation test
             methods

       C.     Development of tissue residue thresholds        .

       D.    Development of trophic transfer models for sediment mediated tissue
             residues
                                        350

-------
VH.  Remediation Methods

      A.    Development and validate methods for in situ containment and treatment
             of contaminated sediments
         i.'       '                        * .                  ''         .
      B.    Develop methods  to  examine rates of natural  recovery  for  benthic
             communities                                      .     ,
Vin.  Ecological Risk Assessment

       A.    Develop methods to integrate stress response relationships for ecologically
             relevant endpoints
                                         351

-------

-------

• • '"• 1
o o -g zr
O O CD C
3 '-a- 5 3
fr\ *-•*• •"{ — j
til's
ฃ CD" 5' ir
2- Q. s? CD
CT. CD m
3-ง o E*
ฃ} vw o --^
Z3 03 ^
0 Q.ง
.-• CD i..
3 ^
<. 03.
— t m
0 CL
1 ; .. ' ID -• • '
ID 0>
la
•-*• 15
m Z5
= CD
OX
CD
CO
^^^^B
.' : '
'I 1 ^
C/) ^^> J^ J1D
CD 0) ง• ^o'
QL c/) ^ X
3 CD ;=i .r-s
^ en 5; Q
CD C/) *<. O
5- "0 _s 5L
C/) O ^ O)
0 CD 0 "
'3 -5 51
.ง ป"i
C — =
Si 3 en
S"'T3 CD
P 03 Q-
CT O^ CD
S R c/T
J/T ง
< QD
|l

CD Bi
•" CD
03 Q.
13
O
O
•• .'• 3- '•
ST
3
- ' - ,5" "V
s.
CD
Q-
^B ^Hk ^^^ ^I^B

C/) ^M
CO-
•••••
CD 5"
, .' ;,3 '-, . .
CD
3
33
CD
0)
CD
0)
                                             o
.  353

-------
1 Planning System
^3
CD
CO
CO
CD
CD
CO
JO
Q
^•^••^•1
DC
O


CZ
CD
- G
O
Q.
E
o
O
o
"of



c.
co
CL.
O
"CD
CD
to
CO
Q
DC
O
1

CO
CD
"o
o>
i_
00
CD
13
CO
p
CD
CO
CD
DC
I

•
CO
CL
JZ
p
CO
CD
CO
CD
DC
CD
.13
CO
JO
1




co
Q.
CO
CL



•
CD
c:
CO
CL
CD
CO
Jฃ
I

•

CO
Q.
; 13
O
O
"c
CO
CL
1

"o
o
o
^•••^
O)
CD
2
0)
p
CO
CD
CO
CD
DC
1

CO
0

4.1
•4— ป
ฃ
E
o
o
o
CO
CD
CO
CD
DC
1

354  .

-------

1
C/)
3"
"2.
CD
.
CO
CD

a
T3
' ^^
Z3
2.
13"
CQ
"O
3
O
CD
CO
CO
';



1
promote g
communfr
ซ
CD
Si
CD

CD
o5
2-
6"
Z3
fj.
zr
^i
CD
CO
0
CD
—5
^mf
0


1
integration
process
^••^
5
^^••M
CD
CQ
CD
O

'••"ง"
Z5
3-
13
CQ






manager™
CD








• . •







1
o"
Q.
O
13
j*^
Q.
CD
2-

13
^
CD
^•^
13
o"
m
^0
^






1
|
"T"
3
Q.
PO"
CO.
•D
~~\
O
o
zr








•


i -
-L o
O .^
o CD"
c o^
ง s
IT

co"
•^•••B , , .
CD

-------

-------
Break-Out Workgroup for Freshwater Sediment Issues:
Overview of the Day
Gary Ankley, U.S. EPA Environmental Research Laboratory - Duluth, MN
I.      General Discussion Format

       A.    Brief presentation of survey results on culturing and testing
             1.     Hyalella azteca
           •  2.     Chironomus tentans
             3.     Lumbriculus variegatus

       B.    Discussion of major culturing and testing issues
             1.     List of proposed issues (by organism)
             2.     Resolution (if possible) of issues

ri.     Selection of Test Species

       A.    Current and historical technical acceptance of test organisms
       B.    Logistical concerns (e.g., organism availability)
       C.    Availability  of some test methods                      .       "

HI.    Survey Techniques

       A.    Questionnaires on culturing/testing distributed to workshop participants and others
             testing target species
       B.    Focused on IL  azteca.  C.. tentans/riparius.  L^ variegatus. but other species
             identified as well
       C.    Results summarized by issue

IV.    General Culturing Issues

       A.    Substrate
       B.    Genetic drift/stream differences                                        '
       C.    Density
       D.    Known age systems
       E.    Water         ,          ,                       •
       F.    Nuisance organisms
       G.    Flow-through vs. static             -     . .
       H.    Light/photoperiod
       I.     Feeds/feeding
       J.     Temperature
       K.    QA/QC (e.g., reproduction, reference toxicants)
                                         •357

-------
       General Testing Issues

       A.    Test lengths/endpoints
       B.    Organism age
       C.    Water renewal (volumes, frequency,'method)
       D.    Physical test system (sediment volume, etc.)
       E.    Test condition and design (chambers, lighting, etc.)
       F.    Interpretation of sediment variables (e.g., organic carbon particle size) on test
             results
       G.    Feeds/feeding regimes
       H.    QA/QC (for acceptable test)
NOTE;  For discussion purposes, the proposed issues initially were ranked based upon:

       • Immediacy of issue                      ,
       • Generality across tests
                                          358

-------
SURVEY RESPONDENTS
Institution
Columbia FWS
Athens FWS
Duluth EPA
University of WI-
Superior
NOAA
(Ann Arbor)
Wright State
ABC Laboratories
Environment
Canada
EVS Consultants
Region 8 EPA
Old Dominion
Cincinnati EPA
Region 1 EPA
University of
Mississippi
Michigan State
University
Maryland DE
Canada Oceans &
Fishes
Miami University
Washington State
DE
H. azteca
X
X
X
x

X
X
X
X
X
X
x
X
X
X
X
X
X
X
C. tentans/riparius
X
X
X
x
. • '
X
X
x
X
•>

x
x
x
x




L. variegatus
X

x
x
x

X
1


-
' • ' 1






.
          359

-------

-------
Development of a Standard Protocol for Testing Hvalella azteca
Teresa Norberg-King, U.S. EPA Environmental Research Laboratory - Duluth, MN
I.     Summary of Culture Methods Used as Reported in Questionnaires


ri.    Summary of Test Methods Used as Reported hi Questionnaires


HI.   Proposed Key Issues for Discussion
                                      361

-------

-------
RANKED ISSUES FOR CULTURING H. AZTECA

       •  Known Age Systems
       •  Feeds/Feeding Regimes
       •  Water (Reconstituted vs. Surface Waters)
       •  Flow-through vs. Static
       •  QA/QC (e.g., Reproduction, Reference
         Toxicants)
       •  Genetic Drift/Strain Differences
       •  Substrate
       •  Density
       •  Nuisance Organisms
      ' •  Light/Photoperiod
       •• Temperature
       •  Other	——
                       363

-------
RANKED  ISSUES FOR TESTING H. AZTECA



     • Test Lengths/Endpoints
                                   [••

     • Organism Age


     • Water Renewal
         (Volumes, Frequency, Method)


     • Interpreting Effects of Sediment
         Variables on Test Results
           (e.g., organic carbon, particle size)


     • Feeding in Tests

     • QA/QC (Criteria for Acceptable Test)


     • Test System
         (e.g., Chamber Size,
         Construction, Replicates,
         Temperature, Light/Photoperiod,
         et cetera)                     ,

      • Water and Sediment-Quality Monitoring


      • Sediment Volumes (Physical test system)


      • Other	—_
                      364

-------
Frequency of Culture Re-starts

  1 month           1    '' ,
  2 month           6
  3 month           1
  4 month           1
  continuous        1
  quarterly          2
  2x/year
  variable
              365

-------
  Variety of Foods Tried (listed singly)

                yeast
              Cerophyllฎ
             wheat grass
               Chlorella
           diatom (Spirulina)
             alfalfa grass
              Tetraminฎ
              Nutrafinฎ
                YCT
             paper towels
           S. capricornutum
           Ankistrodesmus
             maple leaves
paper towels innoculated with Tetramin
             rabbit food
             brine shrimp
               astroturf
            aquatic plants
              sediment
                  366

-------
        Feeding Frequency in Cultures
Flow-through cultures
Static cultures
                  3x/wk       1
                  1x/d        1
                  1x/wk       1
                  Ix/mpnth   1
                  3x/wk       3
                  2x/wk       8
                  1x/wk       2
                  1x/day      1
                  2x/day      1
                  as nee      1
                       367

-------
Waters Used for Culturinq
 tap            7
 well            4
 surface        3
 reconstituted   3
 mix of sources 1
           368

-------
                  Foods fed Cultures of:
Laboratory
Food(sl> Using
                            How Long
ABC Lab.

Dept Fish/Oceans
Environ. Canada
EPA-Duluth
EPA Region 1
EPA Region 8

EPA Newtown

EVS Consultants
MD Dept. Env.
Miami Univ.

Mich. State
NFCRC-Athens
NFCRC-CoIumbia
Old Dominion
State of Wash.
Univ. of Miss.
Univ. of WI-Sup.
Wright State
Yeast, Cerophyllฎ, Chlorella,
wheat grass, diatom, alfalfa
     Tetraminฎ flakes
      Nutrafinฎ flakes
      YCT & diatoms
       rabbit pellets
      paper towels &
       flake fish food
       Cerophyllฎ &
     S. capricornutum
 YCT and S. capricornutum
     Tetraminฎ & leaves
   digested paper towels  '
  inoculated with Tetramin
         Tetraminฎ
          leaves
     leaves & Tetraminฎ
    leaves & rabbit chow
     rabbit food & leaves  .
    leaves & rabbit chow
   YCT & Ankistrodesmus
   rabbit pellets, Tetraminฎ
                          5-6 y
                           ~1 y
                           ~1 y
                           ~1 y
                           ~1 y

                           3 mo

                            2y
                           ~3 y
                            2y

                           1.5 y
                            5y
                            5y
                            2y
                            5y
                            3y
                           6 mo
                           4 mo
                             369

-------
Reconstituted Water
 • 7 labs have used it
  4 with good success
         370

-------
Characteristics of Culture Water (n = 18)
        very soft             1
        soft                  5
        moderately hard      7
        hard                 4
        very hard            1
                    371

-------
Types of Water Renewal in Cultures
      flow-through               3
      static, static renewal        14
                372

-------
Culture Records Desirable/Maintained

   Parental survival            56%
   Age brood animals started   50%
   Routine chemistries         94%
   Quality of food              69%
   Freq. of culture initiations   100%
                  373

-------
Reference Toxicants Used (n = 14)
   cadmium        6
   copper          3
   KCI             5
   NaCI            1
   zinc             1
   chromium        1
                 374

-------
Types of Substrates Currently in Use

  plastic mesh                   2
  gauze                         4
  nitex                          1
  sediment/towels                1
  towels                        2
  sand/towels/nitex               1
  plastic/leaves                  1
  leaves                        4
  mesh/towel                    1
  none                          1
                  375

-------
                     Substrates Used
Laboratory	Choice           Others Tried
ABC Lab.             nylon mesh         maple leaves
Dept Fish/Oceans     sterile gauze     aquatic plants, none,
                                    nitex, leaves, astroturf
Environ. Canada         gauze      .      sediment
EPA-Duluth              gauze         leaves, sediment
EPA Region 1    plastic mesh pad, leaves   leaves only
EPA Region 8          sediment
EPA Newtown       kraft paper towel          leaves
EVS Consult, silica sand, leaves, nitex cones   gauze
MD Dept. Env.           leaves
Miami Univ.     paper towels (unbleached)       -
Mich. State   gauze, unsterilized towel strips
NFCRC-Athens           leaves
NFCRC-Columb  leaves & 3M plastic web        -,
Old Dominion            leaves
State of Wash.           leaves
Univ. of Miss.            leaves
Univ. of WI-Sup.          gauze                -
Wright State      leaves, paper towels
                  polyethylene mesh
                            376

-------
   Culture Temperature (ฐCV

15  •' .  •'.'••'   .•.;•;•".'  '••',' ..;;.
20                              3
23                              8
25                              4
19-23                           1
                377

-------
     Culture Chambers Used
Aquaria Sizes      1 L - 39 L
Water Volume    0.75 L - 38 L
               378

-------
   Test Lengths

96 h             3
7d              4
10 d             8
10-14 d           1
14 d             4
20 d             1
28 d             4
        379

-------
        Test Age

known age             7
sieve for size/age       8
mixed age             2
unknown              1
            380

-------
 Water Renewal and Frequency


Static
      no water replacement        10
      top off                       2


Renewal

      4-6 h               1
      24 h                2
      72 h                1
      1.5ad.dn/d          1
      4addn/d            2
      2x/wk              1
      not specified        1
                381

-------
FEEDING IN SEDIMENT TESTS

  7x/wk                    5
  3x/wk                    5
  2x/wk                    2
  1x/wk                    1
  every 48 h                1
  initiation only             1
  none                    1
             382

-------
Test Acceptability Criteria


        Survival


    60%               1
  i     "         '
    70%               2

    80%              13


    90%               1
           383

-------
What is Reasonable to Evaluate
      Test Acceptability
 Survival
                   yes  18
                   no    0
 Minimum growth
                   yes   3
                   no    8
 Reference Toxicity Test
                   yes   7
                   no    7
                   may be 1
              384

-------
Test Temperatures (ฐC) (n = 17)

  20                     6
  25                     5
  23                     4
  20                     1
  20-25                  1
               385

-------
?
cu
g
LL
O

CO
UJ
O
CC
D
O
CO

CO

O
E
      2
      co
      •o

      I
      CO

      00


      CO
      D
O
   0)
   13

   GO

   CO
         u
   in ฃ,
   ฃซ
c

I
CO
o
CD

      CO
   CO  C

   •=  1
   co  E
O-'>
O c
Q UJ
 C
"5
CO

1
cc
 (0
"5
 o

         _ o
         s>  •
CO
c
o
o
c
V)
CO

J>

o
03
tc
.ฃ to
CO
CO
0)
22 {^
00
CRC-Columbia 4
u_
Z

O
ฃ
W
CO
ฃn
3
(0
C
o
o

CO
LU



CO
CO
C Laboratories 4
CO

















o
en
Dept Environ. 1
•o
c
CO
















r*.
00
\IFCRC-Athens <
bซ

















o
G)
Old Dominion 1


















0)
00
of Mississippi 1
^
•^
                                    o
                                    ฃ
                                    c

                                    o

                                    0)

                                    1
                                    UJ
    o
—   ฃ
ilis
^oo 22-^
     en 0>

ssii
'5> '5 ^ co
0) 0> —* ^j
cc cc •= ฃ
rf rf S .S*
^i ^ CO ซc
Q. Q. ^ >
UJ UJ 2 >
                             CO
CO
         UJ
                                 386

-------
Development of a Standard Protocol for Chironomus tentans
Robert Hoke, AScI - Duluth, MN
I.     Summary of Culture Methods Used as Reported in Questionnaires


n.    Summary of Test Methods Used as Reported in Questionnaires


ffl.   Proposed Key Issues for Discussion
                                     387

-------

-------
                            SURVEY RESULTS

                Ckironomus tentans (riparius) Culture Conditions

                           No. of Responses - 8 (2)
CULTURE TYPE

TEMPERATURE

LIGHT QUAL./INT.

PHOTOPERIOD

CULTURE CHAMBER SIZE

CHAMBER WATER VOLUME

CHAMBER WATER RENEWAL RATE

NO. OF CHAMBERS

NO  OF ORGANISMS/CHAMBER

CHAMBER RESTART INTERVAL

AGE OF RESTART ORGANISMS

ORGANISM REMOVAL

FEEDING REGIME


CHAMBER CLEANING

AERATION

 CULTURE WATER
FT-2.SWR-8

10-25ฐ c

ambient lab fluor./50-120 ft-c

16L/8D

1-40L

1 - 30 L

once/day - evaporative loss only

4-40

,50-800

2X weekly - every 6 months '

 egg cases - <24 h old larvae

 daily - as needed

 bleached-l/unbleached-7/sand-2
 (paper towels)

 none - weekly

 yes-10    "       .•  / ;  .

 soft/moderately hard/very hard,
 lake-l/tap-3/well-5/recon-l
                                      389

-------
                            SURVEY RESULTS
                  Chironomus tentans (riparius) Test Conditions

                            No. of Responses - 8

TEST TYPE      .                      S(6); R(3)-(4/d, 2X or 3x/wk)

TEST DURATION

TEMPERATURE

LIGHT QUAL./INT.

PHOTOPERIOD

TEST CHAMBER SIZE

CHAMBER SEDIMENT VOLUME

CHAMBER   OVERLYING   WATER
VOLUME

NO. OF CHAMBERS (REPS/SAMPLE)

NO. OF ORGANISMS/CHAMBER (REP)

AGE OF TEST ORGANISMS

SIZE OF TEST ORGANISMS

FEEDING REGIME

TEST CHAMBER CLEANING

AERATION

TEST WATER
TEST ACCEPTABHITY CRITERION
 MINIMUM SURVIVAL - 7
 MINIMUM LENGTH - I/WEIGHT - 7
20(2), 22(2), 23(2), 25(2), ซC

ambient lab fliior./25-120 ft-c

16L/8D

50 ml(2), 250(2), 300, 1000(2), 2000

10-200 ml

40-1800 ml (1:4, S:W, 6 of 8)


2-15; 3-4 (6), 15 (2)

15-80

0-16d; 10-14d (7)



daily (5), 2X-3X weekly (3)

none

yes(6), no(2)~ERLD,UM

soft/moderately hard/very hard,
lake-l/tap-3/well-5/iecon-l

>40% saturation (2), temp. (1)
70% (4), 75-80% (1), 80% (2)
                                    390

-------
Chironomus tentans (riparius) CULTURE CONDITION ISSUES
 4-   SUBSTRATE





 4   CULTURE WATER .





 *   FOOD/FEEDING REGIME





 4-   QUALITY ASSURANCE/QUALITY CONTROL
                        391

-------
   Chironomus tentans (riparius) TEST CONDITION ISSUES
4   ORGANISM AGE





4   TEST DESIGN (TYPE, RENEWAL, FEEDING)





+   EFFECTS OF ABIOTIC FACTORS





4*   TEST LENGTH AND ENDPOINTS





4   QUALITY ASSURANCE/QUALITY CONTROL
                        392

-------
ism** fnr Lumhriculus varie^atus
Peter Landrum, NOAA Great Lakes Environmental Research Lab - ^n^or- MI
Gary Ankley, U.S. EPA Environmental Research Laboratory - Duluth, MN
I.    Culturing
      A..    Substrate
      B.     Density
      C:     Water
      D.     Feeding
      E.     Temperature
      F.     Light
      G.     QA/QC
 H.    Testing

       A.    Age
       B.    Loading rates
       C.    Test lengths
             1.    Example
       D.    Water renewal
   -    E.    Sediment volume
     ' , F.  '  Interpreting effects of sediment variables
 HI.   To Feed or Not To Feed?


 IV.   Sediment Avoidance - Effect on Exposure


 V.   Gut Purging for Bioaccumulation
                                        393

-------

-------
m
CD
CD
erg,
CD -3
0CQ
C^CQ
Q-3
CD
I3_
"S
CD
"^ .
o
0
CO
|L
'• 31
o
c
o
o
^^*

_1
CD
O.

c3
JD
13

CO*
v 3-
CO
o
03
"n <
O 03
c ^ CD"
CO *^*" ^
CD 5" • •' i
^l 1
I' z
CO 03
B CD"
o ^
^ CD"
B ^
CD
-Q
E.
ซ$"
co"
' 1— 1-
CD
3
^.

co"
c
CD
O"
o
t— ^ ,
•' • .
•ปJ
03

CD
cr
CD
CD
o
CD
3
C0_
i i
3-
. c-
CO
I-*
cr

o"
^
CD
O
CQ
(_t.
o
3
ง•
55"
13
^
i— t-
CD
— .
C
=
ซ— ป•"
^









O)
c:
cr
CO
1—4-
•^
B
CD
i
CT
O
13
T3
"ง
CD
I— 4-
O
CD
CO
CO
O
<ฃ"
Q.
4^,
OO
IT
























O

^
-JQ
^
Q
[—
<
13
-'Q
B
c
CO













395

-------
ty


Q
LLJ
H>
z:
h-
2
O
o
o
•'•JP
2_
o:
~~^
b
ID
O














o-
•
CO
CD
O
CD
CD
tฃ
C
~05

1
.9
"CD
CD



5




"m
% \*
o
ts
^^
m -
ป w
Q.
•*— >
O
C
CO
E
CD
•*— •
CO

CO
CD
O)
05
0


aboratory
^^^m
^— >
C
CD
IQ
OS
•o
CD
E
^•M
o
CD
CL
CO
,CD
g
"co
o
c:
T3
g

CD
CL
O
*—>
O
.0.
O)
LJ













O
o
CM
C\J
CM
i
CD
1
CD
CL
CD


'ate; reference toxic
mn^
D)
_C
5
1*
0
•o
c:
,g
"cS
CL
0
CL
^^
"o3
O"
0
i—
S

o
E
O
O
Q
a ,
396

-------
CO      S*     :.'•'-' *H        o

CD       JD           CO        CD   CQ

ง=•   -u CD    co 3-3 ^  • 3 g.Q:    ง
3    CD ^    g CD g r-   CDCQ^    ง
CD    ZL      9r. >< ซ<; CD   =r 0> ^    w
ง    -a-n    ^S^ 3   5"^       ^
 •    ^  -U    3Q_Q-(Q   g-g-"    =
     ^^    CD  3; CD  -—.    -

&•   ti    3t.8-r-lsi"-s
3        CD    CD  O~ n?      2 pr ,      '
     o_—   o__=-—- rn    CD x      ^
 ?    8  '    R. 5 Q. 5
 1   ,  ^3      ~J —^ "~" f^

 y>    < • S?   ^ s 5 e
ฐ   rn"      C  — CD  O"    S'3 g    Q.
            , si. CD    *"
     O      "3  -T
        ^^N   •"••   '
     <^ K   CQ ซ<

c   CD CD    ^-H -^ ^. ฑ=    _=;•>->         _j
—*•   Q- ^   3"" ' ^J 3    rT 3 S         "~1

5-   o ฐ    -2-Qs-c    x CD ฐ         rn
y-   ^      -ซ/^5 Q  ^r    ^- ^ CQ         C/)
3' T' vt S   ._*$ฃ,   ••??•
.7; .-^ — x    ^T CD  '    ฃT
 S   ^ป    CD^-S   Q.^        H
 3   (Q 55   S g 05-        .,
      CD      T75 —• ^N -^       CD ^ J        ^-
 g    =3o g -g-^.   CD  o-a-       . o
 3   -'CD   23sc-m~   w.9:^         _
 o    =: CD   c en w S.   5T 5 5-        Q
 S29-   3 g 0-8    .-,2^         Q
 5    S CD    S"i:.^e   ro

                ฃD
                —• i\ ***    l>^
                =•ฃป Q.   O ^3 ^         :e=

                             — ^i-         O
                             —ri __         	i

                             CD  ^         H

                             CD  O         <^k
      rr^ป         *^^ ^  •"*"    *~r" O  ^         v^

      ^ 3       CD 3  CD    05-3         "Z.
       •  —      • XX ซN   .    	_J  J-ป         ^
         ^J.      CD 3  "*    CO ^3

 5"   ^-5T     3'1-g    S- sJ

 
-------
Q
LU

O
O
CO
z:
o
H
Q
z
O
o
o
CO
LU
       12
       CO
       CD
      o:
       CO
       CD
 CO
_CD

JD
       cc
       C
       CD
       E
        CD
       CO
       co
       •*—*
       o
       CD
       M—

       LU

       'o

       "•P^
       CD

       Q_

       fc
       "c
              5 03

              Si
       CO    —
                 CO
                 o
              Q. CD
              CD -^
              vl •ป-•

             T5 "S

              "5g
              P  i
              .I'g
              s  w
              CO  r-
              ^O
              0€
                .  CO
              t-  O
                       T5
                       CD
                        CD

                        O

                        c
                        CD
       >•ป CO
       CD E
       > _CO

       •- co

       1 E
              CD O    —
              9 O     CD
                        O
                 Q.
                 CD


                 "5


                 O
                 CD
                 H-<
                 CO
                 E
                 CO
                 "c
                 co
                 E?
                 6

                 c\i


                              CO
                              T3  CO
                              D)
                                     0)
                              •<-ป CO  CD
                              p .E „_
                              b= c  O
                              __) U-  /HS
                              co co .2
                               ^ .

                               0  >*ง
                              JZ JZ ^
                                 .0) CO
 co
 CD
 O
T5
                              O
                              Z3
                              T3  CD
                              CD  ฑ
•ganisms wi
sediments
                              CO
.l-s
•S  +=;
 CO  jo
    Zi
    E
    o
 o  2
 O  CO
en
                                               Q.
                                               ZJ
         JD
         _CO

          O


          E
          CO
          "c
          o
          E

          CO
          o
          *-•
          CD
sens
CD


CO

CO
arieaa
                                                   CD
                                                   CO

                                                   CD
                                                   CO
 F  E
                                     398

-------


CD
X
posure.















Q.
C
CD
o"
— 4i
0_
en
o
03
Z5
—5
^D

'c
o
CD

5
(Q
CD
f— ฅ
0


•
ciT
CD
g.
CQ
CO
03
Q.
ฃ~
3
5"
03
^^
ซ-*•
O
, c
ซ— h
. CD
O
ZT

•CQ


03
O
O
1
C
•a'
o
-13















O3
Feeding Select
•^
o
03.
13
O

Q3
>**
Z3
CQ
CD
"T*
CD
CD

C?
0


O
O
ntaminated mat
CD














po
Enhanced elim
*•••
03
5"
Z3
^^
<
^^^
V^r
Z3
CD"
Q.
•
'13
O
0
Z3_
Q?


-g
rtitioned materh










.c
13
contaminated 1
•MIII •
O
Q.
03
13
Q.
O"

',


•
Reduce exi
13
CO
c.
CD


^y
w^
0
m
X
CD
O
CD
Q_
3
o
CD
Q.
CQ
• •

-



i:cE
.C' 3"





O"

O

-g
CD
— h
CD
-^




CD
~t __.
QD — ^
S. 3
CD o
03 CQ
3 03
Q- 13
ft
Hฎ
O JD

CD — y
CD

Q.
O
W
CD
03
Q.
CD
^•^
oT
ฃ+:
<"
CD
zz*
^M*
' • a
CD
Q_
3"
o
Q,
CD
CO
c .

CD Eฑ

O-
9^ cF
5. CD
•*•"•ซ *i^
o Q.
-3"
O CQ
03
\j
0 0
13 13
                                                ~n
                                                m
                                                m
                                                Q
            399

-------
CO


i
a
 03
O

2

o
DC
DL

H-


O
the intestional tract and

is
CD
•t— ป
O3
r*"*
M—
O
CO
1—
~*
OD

"5
CO
CD
CO
CO
o

tissues.
>^
"D
o
1
••w
le compounc


'1
o
•
,g
"S
c
"E
"o5
tional content of the
xachlorobiphenyl and
)pyrene body burden
CO
o>
_"c
CD
-ฃZ
•*-*
en
eterminationi
T3

O
"•*— ป
CD
.E
E
0
LL
CD
ฃ
CD
.0
03
•*— *
O
CD
•4-1
CD
T5
0
ฃZ
CO
03
<:

E
CO
03
E
o

O3
'o'
N
CD
CD
•Jk*M
15
o
CM
O
H —

^^
B
c**
>1
03

% difference was not
*^
o
CM
CD
i-
•
fh
Vi/
CO
O
Q.
X
CD
15
CO

Cu
"O

ฃ
03






significant.
_>.

"03
O
to
"ง•
"co

CD
vP '
0s-
0) .
0
ซM^BB
1
O
LO "
CD
^_^
O .
>.
T3
Z3
H— ป
CO

assimilation
CD
4— '
03
O3
vw
Q.
CD
CO
E
o
LL
O
*—
CO
t— ป
CD
4— •
O
o
03
C
g
1JI3
CO
o
[c
E
o
MM*
'1
CO
03
CD
^
^^^
JD

>,
O
.Q






CD
C
CD
/^
CL
'oT
^ **
, N
C
CD
JD

CD
•4—
0
0s-
CO
T3
OS

C
CD
Q.
O
^MM
O
03
X
CD
CD
ฃJ

^
O
O^'
00
CD
CM
O)
C
Z3
Q.
O)
^-
CM
"co
^^^
CD
JD
1
CD
CD
>^
LซLป
'oT
>^_^-
N
C
CD
JD

                                     400

-------
                                       TJ
                                       <

                                       CD
                                        O
                                        O
                                        o
                                        Z3:

                                        CT
                                        P
                                        . -^
                                        CD'
                                        (Q
10
O5
CO

•o
•o
3
O
O)

TJ
T3
 CD
 K)

 T3
 T3

.3
C5
O3
     401

-------

-------
Development of a Standard Acute Amphipod Protocol
Richard C. Swartz,  U.S. EPA Environmental Research Laboratory -Pacific Division
I.     Generic .Protocol Design


n.    Generic Technical Issues


HI.   Species Specific Mo
-------

-------
                 M. Redmond


_0.
o

w
*


.u.
ro



CO
fฐ


ro

ro


CO
0




00
ฃ
o
o
(/>



CO
CD
CO
1

en
ro
o.. ;


ro
CO
CO
i

CO
en


ro
3
O

*""
U9
3

ro
o
D

•ซ*
CO
o
o
D
!-*•










ro z
w ?
0 *—
0 S
03
m
33
m
o
O
^c
m
ro 33
ฐ0 m
0 o



^ 0
ง' ฐ
D) O
't. g
1 ซ
Q. w
3"
3.
^
. 5
?-*
nr
CO
o
fi)
Q.
i:
•o
o
o
"2.

o'
of


S"




.' H
m
ฃป
ซ3
m
33

H
33
m
>
0
u>
n™
s
• ^^™
H
X



            Q

            (D
405
  DRAFT
t>0 IOOT  CITE

-------
                                       M. Redmond
            LIFE CYCLE AT 25ฐC
                 METHODS
•  Newly-released juveniles isolated from
   brooding females in seawater, then held 8-
   10 days
•  10 juveniles/replicate
•  25ฐC, SOppt, 16 hrs light
•  Fed the flagelfate Pseudoisochrysis
   paradoxa
•  Flow-through system, 1 VR/day (seawater +
   food)
•  3 replicates sampled/week for 7 weeks
                                     DRAFT
                                   X>0 WOT CITฃ

-------
                                                 M. Redmond
             Percent mortality, mean +/- sd
           -i  ro  to  A  oi  o  ->4  co  co  o
           o  o  o  o  o  o  o  o  o  o
     ro
     o
CQ


-------
                                               M. Redmond
             Length in mm, mean +/- sd
                              -CO
CQ
 0

 O
 —h
 0)


"O
 =r

•5*
 o
 Q.
 to

 5*

 a
ro
o
    cn
    o
        CO


        I
        I
        3
        m
        CO
            m
            a
            6
            I
                        408
                                            D!
                                              fOOT C|Tฃ

-------
                                                                 M. Eedmond
      o


      to
2
 ฃr   ง,
     *<^

 
            TI
            m
                                         o-  • -s     ^
                                         .=*   o     v^
                                                         Q

                                                         V)
30
     *>•

     O


•<••••


•ซ•••
<ซ•ฃ.
1
3t
o"
(0


•••••I


Ovigerous fe
stage brood)
Bi.


•s?
3
i
o
o
3-
w
o"
a
0
.S
t>
-0 ฎ

(5*
(D

0
(0
s*
CD
Q"
I
(0
                                              ^^



                                              u)





                                              ฃ
                                              (Q

                                              (Q
                                              ง
                                              a
      o


      o
      o
      o
o
r
m

O
rjo
0)
m
30


i

O
z
0)
                                    sr
                                    (Q
                                    Q

                                    cr

                                    5
                                    o
                                    Q.
                                 .409
                                                               MfiT

-------
                              M.  Redmond














f
1".
ฃ
*^


CO
CO

W
o
ฃ
r7

s
CD









O
(Q
5T
3
Q.



CO
O>
•
^1
O
o^

of
'
TJ
CD
^ V
a



03
E.
5'
CO
G3
*<ป


"si
CD

ฐ
O
'(/)
Vr
•^
5'
o^
-K
Q'
Q.





O
.3
(Q
2L
Q)



O>


O


5? .
at
r-
O
C
30
O
m





c/>
m
2
TV
H

^o
lo
™
™n

5'
g
TI
Tl
m
33
m
H
^
9
"O
z:
•o
0
a
0)
0
33
O
m
CO
-z.
D
0)
m
2
5*
_a
m
^L
H
•
0)




^^^5
^j^^
C
33
<
<
T"
^
2]
m
33
o
D
-<
CO
ro
0
o
O





410

-------
                                               M. Reditond
            Length in mm, mean +/- sd
o

o
-+i

ft

13
=r

•5'
o
o.
                       411

-------
                                                 M. Redmond
             Length in mm, mean +/- sd
c> CD
 -A      o  to  ro  to
                                      co co  co
ro
               CD  co  co  ro
                                             co
    O)
 (D

 O
 — *i

 05
 O
 Q.
 to
 Q,
 CD
v-
 to
          1
0

Q.
              CO
II
  ee


  f

    O
              o
  ป


I?

                                         23
                                         O
                                         >
                                         z


                                         o
                                         33
                                         O
                                 DC
                                 D
                                 T1
                                 TI
                                 m
                                 73
                                 m
                                 z
                                 o
                                 m
                                 o

                                 q
                                 m

                                 ^
                                 m
                                 o
                          412

-------
                                M.  Redmond
w
6666


















photoperiod?

















- •—
CD
CD
a
-*
o
— t*
o
?
*-K
0
(Q

V
_J.
<
•^
a
09
-O













shipping/handii
. 3
CQ
•ฐ





,










rt
)w reproduction
09
Q.

o
.
/ft
c^
T3
CD
\mf
I
o

to





Survival curves
O
09
3"
2.
•o
Q.
CD
termine
03
O
O
CD
•o
oT
cr
CD
o
o
3
o
to


a
09
—f
2.
3
CD
™^
O
1"
CD
CD
09
I
g
o
Q.
09
0
Q.

-






7v
3 '
O
3
&
(Q
CD
*E'
O-
CD"
CO
o
3
cr
o
o
s.
S"
Q.
=?
3
o

CC2
O
o
c
" -*
CD
09
O
^^
O
o
o"
•o
0
o
o
o,
(D
CD
CD
3
Q.



„








O
r-ฅ -
&
SL
?
0
c
-•t ' -
o
09
MV
Q.
O
0
3.
^
?5
09
Q.
(/>
O
1
CD
3

(Q
•^ ".
0
r^1

CD
a
0)





^
>
-<












413

-------

-------
                    J. Laitiberson
EOHAUSTORIUS
         415

-------
                                                            J. Larriberson
               EOHAUSTORIUS ESTUARIUS   (HAUSTORIDAE)
Geographic range:



Habitat:


Nutrition: .

Life cycle:

Source of amphipods:


Life stage tested:
Central B.C. to central California
  (other haustorids along Atlantic,
   Pacific and Gulf coasts)

Free-burrowing sand dweller,
upper intertidal to shallow subtidal

Probable deposit feeder

Probably annual, not cultured

Field collected, sandy sediment
  (shovel, sieve, bucket)

Large immature to mature,"' both sexes
                                   416

-------
                                                         3. Larribefson
  ACUTE SEDIMENT TOXICITY TEST CONDITIONS FOR EOHAUSTORIUS  ESTUARIUS
Temperature:

Salinity:

Photoperiod:

Light quality:

Light intensity:


Test chamber sediment depth:

Test chamber water volume:

Number of organisms per
    test chamber:
15ฐC  (5 to 25ฐC)

28 ppt  (2-35 ppt)

Continuous light

Fluorescent lights           •

Normal room lights
   (subdued light  for water only  tests)

2  cm

Fill to 950 ml
20
                                   417

-------
                                                           J. Larrfoerson
  ACUTE SEDIMENT TOXICITY TEST CONDITIONS  FOR EOHAUSTORIUS  ESTUARIUS
                             (continued)
Feeding regime:
Aeration:

Test endpoints:
Control sediment:
Grain size:
Reference toxicant  test:
No food added during acute test
Air bubbled through a 1-ml diposable
  glass pipette
Emergence, mortality, reburial
Collection site sediment, 0.5-mm sieved
92% survival in 80% silt-clay
97% survival in sandy sediments
cadmium chloride, 4-day water only
Mean LC50 = 13.05  (4.38-21.72) **  .
Mean EC50 = 7.07  (1.81-12.33) **
   ** Numbers  in- parentheses are reference toxicant warning limits
         (95% confidence limits ='mean +/.- 2 standard deviations)
                                  418

-------
                                                             J. Laraberson
EOHAUSTORIUS ESTUARIUS. ACUTE SEDIMENT TOXICITY TEST


    ADVANTAGES              ,                          *
        Ease of handling and collection from field
        Salinity tolerance over a broad range
        Grain size tolerance
        Year round availability, can be shipped

   'LIMITATIONS   '          ,                              ,'
        Annual life cycle
             Cannot culture            ;
     •  -•     Cannot use for chronic tests
        Variable response to reference toxicant  (cadmium) _•

    RESEARCH NEEDED
        Factors affecting sensitivity to reference toxicant
      •  Interlaboratbry comparison of test method
        Tolerance limits -  salinity, temperature, grain size
        Field validation                           ,
                                     419

-------
                                                •      Ji- *i
2. Fluoranthene Tolerance:  Eohaustorius estuarius was siigr
 " more sensitive to fluoranthene than Hyalella azteca at 2 ppt
  salinity, and slightly less sensitive than Rhepoxynius
  abronius at 28 ppt.  There was.no significant interaction
  between salinity and fluoranthene tolerance for E. estuarius.
arson
          Eohaustorius-Salinity-Fluoranthene
               10'


Spec
Hya
Eoh
Eon
Eoh
Eoh
Eoh
Rhe
1
a
a
a
a
a
P



i es
el
us
us
us
us
us
1
t
t
t
t
t
a
or i us
or i us
o r i u s
or i us
or i us
oxynius;
20 ฃ-•
FLUOR (PPM)
Sal ini tv
2 ppt
2
5
10
15
28
28

i f2
LC50
21
13
14
15
13
17
^6
.2
-8
.0
.1
.9
.5
.6
•ป•


957. C.L
Upper Lowe
18.
12.
12.
13.
12.
14.
. 5.
1
3
5
2
2
9
9
24.
16.
15.
17.
15.
20.
7.

•
r
8
0
8
0
9
5
4

-------
                                                           B. McGee
        SUMMARY OF 10 D SEDIMENT TOXICITY TEST CONDITIONS
                  USING Leptocheirus plumulosus
1.  Temperature
2.  Salinity

3.  Photoperiod
4.  Volume of test chamber
5.  Water:sediment ratio
6.  Size/age of test organism
7.  No. of organisms/chamber
8.  Overlying water

9.  Negative control

10.  Positive control

11.  Source
 20;  25  ฐC
 Dependent  on objectives of
 the  study
 16:8 light:dark
 1 L
 ซ 4:1 (v:v)
 ,3 -  5 mm
 20
 Synthetic  sea salts;
 natural seawater
 Fine sediment (>85% silt
 clay); salinity ?
 96h aqueous CdCl2
 @ 20 ฐC and 20 %
. Field collected; cultured
                                421

-------
                                                     B. McGee
Research  topics
      •  Effects of salinity on test results
           - Is acclimation necessary?
           - Will the acclimation salinity influence
            test sensitivity?

      •  Sensitivity to common sediment contaminants
           - Expand the chemical database
           - Field validation
           - Interspecies comparisons

      •  Sensitivity differences among sources of amphipods
           - Laboratory reared versus field collected
                               422

-------
                                             B. McGee
Effect of acclimation salinity on survival of
  juvenile Leptocheirus in 10 d exposures
    100
               Acclimation salinity
                 7ppt •i20ppt
                   Test salinity (ppt)
                         423

-------
                                                             B.
             Laboratory Reared Versus  Field Collected
               Amphipods in Sediment Toxicity Tests  .
                       ADVANTAGES
Laboratory reared

Year round availability

Geographic availability

Reared under known,
controlled conditions
Field collected

Subsample of a "natural"
population

Easy to obtain large numbers
with minimal effort and cost
                       DISADVANTAGES
$ Genetic effects
(e.g., inbreeding, selection)

$ Influence of culture
condition dn test sensitivity

Cost  (time & money)
Limited availability (seasonal,
geographic)

$ Seasonal &/or geographic
variation in sensitivity
$ Potential research topics
                                424

-------
                                            R. Alden
Lepidactylus dytiscus Distribution and Ecology:

    Lepidactylus dytiscus is broadly distributed
throughout the upper Chesapeake Bay and its
tributaries to Florida. It is an intertidal species
found 1-2 meters above the low tide mark in moist
coarse sand in summer to approximately 1-3
meters below low tide in winter months.   L.
dytiscus burrows freely in coarse sand throughout
the year generally burrowing to 4 cm in summer
and 6-7 cm in winter. Densities can be 1200-1500
individuals/M2.  Average densities are
approximately 150-200/M2 where L. dytiscus
occurs.

     Bousfield (1970) reports feeding in L. dytiscus
is by suspension, although it may supplementarily
deposit feed. Lab held animals  are fed small
amounts of Anemia salina and algae.

     Reproduction generally occurs spring through
 fall with large females  overwintering. Bimodal
 reproduction appears to be the rule with early
 spring and late  summer recruitment appearing to
 be greatest.
                         425

-------
                                       R. Alden
                            CN
                            oo
                             CO
                             03
                             I
                             O
                             N
                             CJ
                             cd
                             4—1
                             c
                             03
                             t-
                            O

                             E
                             o
426

-------
CULTURE CONDITIONS FOR LEPIDACTYLUS DYTISCUS AT AMRL
                                                                          ,R. Alden.
CULTURE CONDITION
CONDITION USED BY LABORATORY
1. Test type

2. Temperature

3. Light quality

4. Light intensity

5. Photoperiod

6. Culture chamber size

7. Culture water volume

8. Restart duration

9. Renewal of culture water

 10. Removal of offspring
     (frequency)
 11. Age of restart organisms
 12. No. of organisms/culture
   chamber

 13. No. of culture tanks

 14. Feeding regime
 15. Substrate used
 16. Chamber cleaning
 17. Aeration
 18. Culture water
Static non-renewal
20 + 2ฐC
ambient laboratory
ambient laboratory
16:8 L:D
20 gallons
 18 gallons
 I vr
50% weeklv
2/vr
 Various
 1200
 arterriia (twice/week)
 native coarse sand
 occasional stirring and siphoning
 moderate w/filtration
 DI w/ artificial sea salts
                                       '427

-------
0)
LLJ
1^"    —m -
 -  a.
     a.
 cc
 O
 O
LLJ
     OCL
^  'X
Q.  LU

X
CL
 R. Alden
 CO
 5
 O
 CO
 c
 s
 •Q
 (0
*co
 (
 3
 to
 0)

 U4
                                                                   o>
                                                                   00
                                                                   O)
                                                                   0)
                                    428
      0)
     O

      E
      O

     uZ
     •Je

-------
REFERENCE TOXICANT DATA:
                                                R. Alden
  r ADMTIJM CHLORIDE TEST:

  At 28 ppt the measured LC50 was 6.33 mg/L Cd (95% CI
  11.22-4.35) with a control survival of 97% (EPA Probit
  Analysis, Version 1.4).

  At 20 ppt the measured LC50 was 6.13 mg/L Cd (95% CI
  7.25 and 5.3), with a control survival of 97% (EPA Probit
  Analysis, Version 1.4).

  Results of reference toxicant tests conducted at intervals
  throughout the year showed sensitivities in the range of 6.3
  to 2.5 mg/L Cd, with a trend toward increasing sensitivity
  in the early fall population.

CADMIUM CHLORIDE REFERENCE TOXICANT TEST
                 (96-Hour, Water Only)


       LCSO's as mg/L Cadmium
12

10
8

6


4


n

















































i
!
:
i
. {
;;




li:

1 1
.'. :
?
i i
I 1
h'


















































i
L










. i

i




















1
!!!
-1 .
; i
I .








; ' ;
Ml
! !
iii


• ..-••-




_ _ —

. . i i ' . . -
' Si •
i I
        Dec. '89  Dec. '89 May '90  Sep. '90  Oct. '90
                     Date of Exposure
                          429
                                         20 ppt D 28 ppt

-------
                                               R. Alden
                TOXICANT DATA:
                 NE TEST:
   An  average survival of 94%  was found in both the
   sediment and the acetone controls.
                                       % •
   Calculations using the Moving Average  Angle Method
   indicated a nominal LC50 of 1.44 ing/kg (95% confidence
   intervals 1.33  and 1.55 ing/kg) and a measured LC50
   value of 0.793 mg/kg (95% confidence intervals 0.698 and
   0.887 mg/kg) fluoranthene.


COMPARISON OF AMPHIPOD SENSITIVITIES
      (10-Day Fluoranthene Exposure, 28ppt)
       mg/kg
    14
           R. abronius* El E. estuarius* • L. dytiscus
*From DeWitt et ai., 1989
                             430

-------
                                             R. Alden
     IALINITY TEST:

    Statistical analysis using anova and regressions showed no
    significant difference in survival of amphipods tested in
    salinities from 5 to 40 ppt.

    Average survival in the controls was 93.3%.

    Test survival averaged over all test groups was 93.3% over
    the 14 day test period, with no less than 90% survival in
    any group.
       SALINITY TOLERANCE
              (10-Day Exposure)
       Percent Survival
    120
    100
     80
     60
     40
     20
      0
ii
WA
>//////
                  10      20      30
                     Salinity (ppt)
                          431

                              40
                                     D Control salinity
NOTE: 95% Confidence Limits Shown

-------
                                                       R. Alden
U
 i
 p
 O
               oo
 CC


U
                               00
Cs
                               oo
                               ON

                                                 O\
                                                 oo
                      U


                      H
                                                  CO
                               432

-------
                         P. Chapman
                             CO
                              CO
433

-------
                                                                       P. Chapman
                                                             cvj
I
 CO
 0)



I
 O

1
                                                                      0) •*

                                                                      as
                                                                      03 i-
                                                                      U-O
= 5.

Si
c .c
r
  0
ca C
ป- o
** *i
= T>
V fl)

om
  o
ซ o
ซ c
i- 0)
ซ >-
o ซJ
O o
  oc
                                                                         0)
                                                                         o
                                                                       0)
                                                                         0)
                                                                        cr
                                                                       o
                                                                       o
                                         434

-------
                       P. Chapman
                         o
                         o

                         1
                          I


                          I
                          sr
                          3"
435

-------
                                                              P.  Chapman
ง
c:
o
CO

ง
                                                           eo
                                     436

-------
                      p. Chapman

                           '
                        CD
                        •S'.
437

-------
                                                                      P. Chapman
 o
.s.
                                                                     a*
                                                                     m ซ
                                                                     •z v
                                                                     CO w
                                                                     u-0
                                                                     o
                                                                      .
                                                                     C j=
                                                                     ซco
                                                                    • •*- >
OJ 
0 O
09 C.
i- 0)
ซ S
O ซ
O ซ
  tr

  0)
  o

  c
                                                                     u. ฃ
                                                                       0)
                                                                       QC
                                                                     c
                                                                     o
                                                                     o
                                    438

-------
Development of a Standard Chronic Amphipod Protocol
John Scott, SAIC - Narragansett, RI
       •with contributions from:
             Ted DeWitt, AScI - Newport,  OR
             Michelle Redmond, AScI - Newport, OR
             Chris Schlekat, Maryland Department of the Environment r Baltimore, MD
I.     Introduction                                        (Scott)

       A.    Historieal background
       B.    Applications of chronic procedures
       C.    Minimum requirements of the methods   ,
       Chronic Procedures Using Leptocheirus plumulosus   (Dewitt)

      1 A.    Preliminary Leptocheirus plumulosus chronic method

             1.     Species selection
                    a.     Culturability
                    b.     Contaminant sensitivity
                    c.     Handling ,

             2.     Culture methods
        '            a.     Physical requirements        !
                -         i.      Tubs
       ';...•       ,     ii.     Salinity
                          iii.    Temperature
                          iv.    Light
                          v.     Sediment

             3.     Test animals                             ,
                    a.     Life stage
                    b.     Number per replicate
                    c.     Feeding
                    d.   , Handling and recovery
                    e.     Responses
                          i.      Mortality *
                          -.ii..    Growth
                          iii.    Reproduction
                          iv.    Others
        ,            f.     Tolerance limits
                          i.      Grain  size
                          ii.     Salinity
                          iii.    Organic enrichment
                          iv.    Seasonality
                    g.     Special requirements
                          i.      Quarantine laws and practices
                                         439

-------
       4.     Exposure logistics
             a.     Chamber characteristics
             b.     Exposure duration                                       •  s
             c.     Sediment depth and volume
             d.     Water change
             e.     Aeration
             f.     Temperature
             g.     Salinity
             h.     Dissolved oxygen
             i.     Light
                    i.      Photoperipd
                    ii.     Intensity and quality
             j.     Other environmental variables

       5.     Experimental design
             a.     Treatments
             b.     Replication
             c.     Statistics
                    i.      Analytical methods
                    ii.     Variability and power
             d.     Controls
                    i.      QA/QA
                           - Performance (health)
                           - Reference toxicant
                    ii.     Experimental
                           - Carrier (spiked sediment)
                           - Site (dilution  series)
                           - Environmental (grain size, TOC, salinity, temperature)
             e.     QA/QC
                    i.      Environmental  conditions
                    ii.     Response criteria (mean, variability)
                    iii.     Controls                              •      .  .    '.

B.     Research findings

       1.     Sensitivity to sediment variables
             a.     Mortality and growth

       2.     Sensitivity to phenanthrene-spiked sediment
             a.     Relative sensitivity of  mortality, growth,  and fertility

       3.     Sensitivity to contaminated sediment dilution series
             a.     Relative sensitivity of  mortality, growth,  and fertility
       4.     Nutrition affects growth
C.     Comparison of EPA and Maryland Department of Environment methods

                                   440

-------
      D.    Research needs
                                                i                 .         ,    i
                                                          /          •               "
            1.     Influence of nutrition on lexicological sensitivity

            2.     Influence of other variables on toxicological sensitivity
                   a.      Temperature
                   b.      Salinity
                   c.      Sediment grain size
                   d.      Other variables

            3.     QA/QC issues
                   a.      Development of reference toxicant method for growth and fertility
                   b.      Environmental conditions during exposure
                      \           -                                  ,
            4.     Relative sensitivity of cultured and field-collected animals

            5.     Simplification of culture and feeding methods

            6.     Relative sensitivity to other species (acute and chronic)

            7.     Experimental design optimization

             8.     Development of toxicological database (chemical diversity)

             9.     Inter-laboratory comparison

             10.   Field validation


n.     Chronic Procedures Using Leptocheirus plumulosus   (Schlekat)

       A.    Characteristics of the test organism
       B.    Selection of the test endpoints                   ._.-.''
       C.    Elements of the procedure                      '
       D.    Representative results
       E.    Research needs


HI.    Chronic Procedures Using Ampelisca abdita          (Redmond and Scott)

       A.   . Characteristics of the test organism
       B.    Selection of the test endpoints
       C.    Elements of the procedure
       D.    Representative results
       E.    Research needs
                                          441

-------
IV.   Discussion                                        (Workgroup)

      A.    Input from workgroup these procedures
      B.    Experience with other species
V.    Summary and Recommendations                    (Scott)
                                       442

-------
                                                            J. Scott
                   CHRONIC TEST METHODS
LEPTOCHEERUS
STATUS:
ISSUES:
GOAL:
•    PRELIMINARY METHOD
          28-DAY SURVIVAL, GROWTH
               REPRO
•    MULTIPLE LABORATORY USE
          MINOR DIFFERENCES
•    CULTURE/FIELD COLLECTED

NUTRITION-DIET QUANT/QUAL
INTER-INTRA LAB VARIABILITY
FIELD VALIDATION
ROLE OF SEDIMENT NUTRITION QUAL
REF TOX FOR CHRONIC ENDPOINTS
DRAFT STANDARD 9/93
     GUIDE?
FALLASTM
 ATVrPRT.TSr.A        ,

 STATUS:   PREUMINARY/SUBLETHAL METHOD
               20-DAY - GROWTH
          LIFE CYCLE 30-40 DAY-DEVELOPING
          CULTURE METHODS - DEVELOPING

 ISSUES:    TEST DESIGN FOR REPRO ENDPTS
          OTHER SAME AS LX,

 GOAL:    DRAFT STANDARD SUBLETHAL -
               GUIDE?  CHRONIC?
                                                  9/93 -> FALL ASTM
                                443

-------
                                    J. Scott
TOXICITY TEST INTERPRETIVE GUIDANCE
      STATISTICAL SIGNIFICANCE
      DETECTABLE SIGNIFICANCE
      BIOLOGICAL SIGNIFICANCE
                444

-------
                                        i Scott
  INTERPRETATIVE GUIDANCE




  DETECTABLE SIGNIFICANCE








Statistical significance incorporates within




    test variability among replicates.






  	but what about test performance




           variability???
                 445

-------
                                     J. Scott
     INTERPRETATIVE GUIDANCE




     DETECTABLE SIGNIFICANCE








     CALCULATION OF THE LEAST




    SIGNIFICANT DIFFERENCE (LSD)






    o Conduct one-way Mest



    o Assume unequal variances




    o Generate a t value.



    o Conduct ANOVA to generate MSE
LSD = ^.0.05 SqRt ((1/NC + 1/N,) * MSE)
                  446

-------
                                                  J. Scott
                               Frequency
o  a.
   3
   JZ


   a
   CD
   i—ซ
   CD

o
O
   cr
   CD

   O
   =ti
   CD


   •1
   O
   CD
            0-5
         5.1-10
        10.1-15
        15.1-20
S 2-  20.1-25
        25.1-30
        30.1-35
        35.1-40
        40.1-45
                              447

-------
 cc
 Ci.
 cc
g
      0
      O

      0
      ir:
                                                      J. Scott
      o  8
      0
      •S
      0
                                            gaSsgsgsfeg^?*

                                                   Is
      c  E o
      co  o E
     O  ฃ -I
   cvj  r^
   m"  CD



ฐls,
'r"  CO  c
 ป  o  co
c ^  a:
                 o
                 CO
                                                         o
                                                                   Q -=-

                                                                   0  E
                                                          -1-'OS
                                                         S 8
                                                         0)
                                                                   Q

                                                                   |


                                                                   1
                                                       9I.-I.-OI.
                                                       OI--I.-9
                                            9-0
o
OJ
                                          o
                                    448

-------
' , . : J. Scott
100-

on .
oU
f\f\
60-
40-
20-
0-
V

EMAP-1990






•-...- "^gg^
, . ฃ*?•ฃ:.'' ~*ฃ?j '•-

u? p

_.j^vซr-


:^S

:'^rSP
to
n=157
range: 2.3 - 32.4%

r^ptpr'tr'ri *i ooo/ -jpttp..-—-- x..—.—

the control 95% of the time.
- Mi^tfes. .- ' • ' - • • . , ;
' 1 : 	 1 	 -t- 	 1 	 1 	 1
S • ซfl • O m oin
o V V 51 • -V 9 9 t T
ui d ui o iri 5 . ' 5 d
• •ซ- •ซ- C\J CM TO CO TT .
100-

c
D '•
05 '40-
i_
u.
20-
0"
~
5

NOAA
Detected a 28% difference from ranae-69 44 oฐ/
the control 95% of the time. 9 * •y"44-u/0






IO O
'6 ; ,.*7 "
.^fa]w2|gป;
" '""P* ' 'J^^ปff^V>
-,-zg?'*'*?'


10
T-
..gl^pgj- . '
''-^vSBJ^^ss ?!* '
SJ^^^^- ' :
.

S !P, 2 "* o 10
CM CM CO COf V
to d 10' o 10 d 10' o"
^ - - •ป- CM CM CO CO V
150-
120-
90-

60 -
30-
~

.-> • , -





••gQ<ฃ^
'
up o
o T

;gf|pi]g

-••J^:Ae^^

'^^&งfe'-
• • ^y/"-ป**?'""
10
1
Both
n=281
range: 2.3 - 44.0%
x^j--- ~-i:i
Detected a 24% difference from
?I8II1I the control 95% of the time.

?- 9 ซ •?' f ?'
^" • T" T* ^— . ป— ' w.
w ? ซ5 - S - ซ o' u? o
                          CM
                                CM
                                      CO
                                            CO
Least Significant Difference (% of Control Survival)
                       449

-------
                                              J. Scott
       INTERPRETATIVE GUIDANCE




        BIOLOGICAL SIGNIFICANCE
ESTABLISH BASIC POPULATION PARAMETERS




      o Age-specific survival and fertility




      o Model-based population estimation



      o Leslie Projection Matrix
               TEST DESIGN




      o Test duration of 70 days




      o Regular non-destructive sampling




      o Examine mortality effects at 10-day intervals
                      450

-------
                                                    J. Scott
                 FINITE GROWTH RATE
       CO
       o>
         CO
         00
o
ro
o>
o
00
m
•n
m
i
CO
H
>
O
m
CO
c
O
;o
CO
o
to
    O
    •
    O)
    o
    • -
    00
                                                         O)
                                                         CD
                                                     ป ซ^
                                                     O

                                                     rn

                                                     fi)
                                                    CQ
                                                     CD
                                                     C/>
                                                     c
                                                     2

                                                     o
                                                     CO
                           451

-------
                                           J. Scott
AMPfflPOD CHRONIC TESTS








  WHY DO CHRONIC TESTS?






    More sensitive than acute




More relevant ecological processes




  Understand population biology



  Determine sensitive life stages



      EPA/COE guidance




  Sediment Management Strategy
               452

-------
                                         J. Scott.
 AMPHIPOD CHRONIC TESTS








       RESEARCH GOAL






o Understand population biology




o Optimize design for:




         information content




         cost efficiency
                453

-------
                                            J. Scott
   AMPfflPOD CHRONIC TESTS
EARLY DESIGNS WITH AMPELISCA

 o Population sampling approach:

      initiate with ovigerous females

      two generations

 o Suspended sediments

 o What we learned:

      amphipods will reproduce in the laboratory
                         r    ' '
      growth is  a sensitive endpoint

      developed crude  population models

 o Limitations:

      high variability

      no  age standardization

      not amenable to model-based approaches
                  454

-------
                                        J. Scott
   MINIMUM REQUIREMENTS
      FOR TEST PROTOCOL
 Responsive to chemical contaminants

         Relatively sensitive

Intra- and interlaboratory variability low

         Organism available

          Well documented
                 455

-------

-------
                                                                    T. "DeWitt
n>
ฅ
 f
 r>
 s:
 w>
                                             ^


                                             i
                                             5?"
                                             3
                                             a
                                             s.
                                              o
                                              3
                                              en



                                              I
                                              s.
                                 459

-------
                                                                      T. DeWitt
       CD

      to

       I
       CD
      CO
b    o
^•^
CO
"ZL
LLJ
CO
LLJ
^>
ง
LLJ
DC
CO
CD
•5
CO
o
CL
X
LLJ
JZ
CD
O)


Q.
Q.
SS
CD
st
3-5
0,
i~o

"c '~o>
15 ^^
CO


CO
CD
"o
CD
Q.
CO


O
CD

CD
t—

in

CO
CM



II
Ampelisca abdita


r>- co
Tt CM
CD CD

CD CM
CO CO
CO CD
1
m if)

CO O
CM CM



Lepidactvius dvtiscus


O) h~ O) CD
i- CD o in
CD CD CD CD

CO O CD fe
r- co o ฐ?
CM' oJ CM $2

o in o in

CO CO O O
CM CM CM CM

CO
3
Leptocheirus plumules
(sub-adults)


CM
CD

a'
CD

in

o
CM



Monoculodes edwards


00 h-
r>- CM
CD CD

5 5!
^ CD

in m

00 O
CM (M



Eohaustorius estuarius


CO
0
0

O)
CD

in

CO
CM



1 Rhepoxvhius abronius
                                        460

-------
                         T. DeWitt
jS60
|40
<^ 20 i
i
0 i
?8'
ฃ6
55 2
0
1
^s'
I 4
2
n

Control
D ,
10-d
28-d 2
0
o A
h 0 0
0
I O O O
l .*0 .A A ,

f 8ง o i o e ง @
- o
1 1 . 'I :
\ •
L o . o -
o o
O . : . . . • '
--,' ฐ ฐ 8 ง -.0.
O ' ..
	 , 	 __i 	 ' 	 • 	 —J 	 ; 	 • 	 i 	
Phenanthrene (mg/dry kg)
           461

-------
                                         T. DeWitt
.-I-80
"560
tr
Sฐ40
^o [
^20 i
(
ni

• Control
D
'. 10-d
28-d
p
O
A
1 A
A A
J- A 0 0
ป * 0
i A O A O
I O O
i i A I 	 1 	 1 	 ' 	 1 	
O
b
A
A
A
1 . 1



1=
0)
25

(
c
6 !
4
1
2
n

> ง
I i 8
e
!•ซ ป
1 1 1



O
8
*
1.1.
•JO ฃL
'^ [1


10



 8
 2


 0
            o
            o
            o
  0
20     40     60     80     100

    % Curtis Cr. Sediment
120
                       462

-------
                                                               T. DeWitt
  15
•=10
CO
tr
o
                                                   Newborn
                                          Newborn
                        SubAdult  SubAdult
                SubAdult
        SubAdult
                                Newborn
          No       No      No       Yes      Yes      Yes      Yes
           No      No
No      Yes      Yes'     Yes

   Food Added
                                                               Yes
                                     463

-------

-------
              c.
               Schlekat
 c
 9  , -
 a 9V
 s.si.

it
 •*1  o
 P  2,
 O  OTQ
 ^  n

 ซi
 o  ^
 --9-"
 & p
 ^ S
;a-a
 ป 3
 W5 ^
   O

   9
       P
       a
    c  ^
             w-
             s
         ป-.  P.
             p
             p,
             -9
             a
       -S
       c
       — sr  ฐ
       s 1  -
       ^ S' " 5'
       oป  ง
             fD
             ซ

             P
H".^
^  s^
   o
   5
   P
   ffi
465
   O


   'W5
                    QTQ

-------
                                                          C. Schlekat
      SUMMARY OF TEST CONDITIONS FOR PARTIAL LIFE-CYCLE TEST

              CONDUCTED WITH Leptocheirus plumulosus
1) ORIGIN OF TEST ANIMALS:
2) NO. AMPHIPODS/CHAMBER:

3) VOLUME OF TEST CHAMBER:

4 ^WATER: SEDIMENT RATIO:

5) PHOTOPERIOD:

6)fWATER SOURCE:



7)"*TEST WATER TEMPERATURE:

8)tTEST WATER SALINITY:



9) TEST DURATION:

10)fTEST ENDPOINTS:
Obtained by isolating gravid 9s
in  control sediment for  <  1
week.   Offspring collected on
250  Jim  sieve are  utilized in
test.  Size has  ranged from 1.2
to 1.5 mm.

20

1 L

4:1  (v:v)

16:8  (light:dark)

Artificial seawater diluted with
distilled  and spring water.    :

20 ฑ 2ฐC

Ambient interstitial salinity
of test sediment

28 d; Static-renewal 2 X week

Survival,  length,  measures of
reproduction
11) POSITIVE CONTROL:
12)fFEEDIN6 REGIME:
i3)tcoNTROL SEDIMENT:
Aqueous cadmium chloride @ 20ฐC
and 20 %o
6-12   mg  TetraMin
Conditioning 3 X week
Tetra
Amphipod    collection    site
sediment  (Corsica River, Queen
Anne's County, MD;
93% silt-clay)
   Different from EPA-Newport
                                466

-------
                             C. Schlekat
S.cg
    ft  &
    S- B
    B: P
       O
       S
P
^ 5  ^
_^. *S  ^
5e  ฎ
ii ซ*ซ  M
3 S"q
'3
'S3
 3
 P
 9
 a
       P
 ft
 Q-
 S*
ere

 3
CTCr
es

     •• C
       P

    ซ  &
    ft  **
    O- <
    n^ซ

    3
    ft
    53
    ซ•*>
    f5
    O
    S
    ซ^
    P

    &

   "S*

    &
    o*
    S

    CfQ

    P
             ?g-ฃ
             ft  u.
                o
                0
                i-*

                P
      ง  J'
      ^*"^  ^C3
      en  "1
      H* P
      ^v  P^^
      c/i  ^
      ^  ^
      a ^
ft2 .   ป—<•
ฃ    s
^^*    ^^^
ijjg^    I"''
      ft' 2
     •5 a
      ซ"5

      งt
      sr ^
      P C/i

      tป
      •5 5
      P Ci-
      ^^ A
      5ซ* M-
      O g-.-
      -B ^
      CTQ &
      p S
               %  O
               ^C ' ^^
               ft  3
               53  ^53
               B: P

               *V P
               P  a
               B  5
               CL ft
               p  ?/r
•<3 rj    vra  --^    >^
   >*•        ป•-•   ^2
ปซ•• C   . .*•• 62     ^
u
                   a
                   ^* **•
                   ง  P
                   s  S
                  .^5  55
                   p  B
                   .ป-<  ซ^
f. sed
                   2..B
                   ป..g.
                       ^*-
                       ซ
                       O

                   ^S
                       P
                       S
                           B
                           P
                           O
                           B
                            off
                                     ft)
                            &  sr
&  5
•OQ  5
n   &
CLCTQ
N^ป
 n   N!
 &

 C/3
        467

-------
                                                                     C. Schlekat
T3
-o -o

O O
W '-i
   Q)  
ป-3
CO
p
CO
o

3
6
j3
3,

to
3
{_


3
"-0
\
cd
o
CO
-4-2
N
fO

(0
ฃ3
CD
   a,  p,
o
                                                                             0)
                                                                            OQ


                                                                            ^
                                                                             CD
                                                                             
-------
                                          C. Schlekat
            to
                   % SURVIVAL
                              cn
O
.o
• GJ
-o
• O
                                                 CO
                                                  O
                                                '  O
H- as co >- TO
?ฐ CO J- C7I 2,
                                         o o n p
                                         c c e o
  oln n
0111
*•( fD >P to
Ct) CD (^ ft)
CD .X" PT 55*
     ^^>^^^^
     <^/^%^^
     ss^^sssss^
                                                  co
                                                  O
                                                  00

                                                  S
                                                  CO
           3

           O

           CD

           (—'•
           CD
                                                  CO
                                                  CD
                                                  a
                                                  t—'•
                                                  3
                                                  CD
                                                  O
                                                  O
                                                  -0
                                                  CP
                                                  ซ-l-
                                                  I—'-
                                                  o
             469

-------
                                                          C. Schlekat
rt
o
•ป— <
-*j
&
6
cti
o
o
-d
0
to

tM
O
 cd
 $-4

O
 0
 J-.
u
 o
•-O

1
 ft,
 0
 o
 so
                   *•
                                                  _ o
              ^^S^S^^^
       
-------
                                              C. Schlekat
                         % SURVIVAL
CO
P3
O
"0
t?d
                     471

-------

-------
   APPENDIX A:
WORKSHOP AGENDA
        473

-------

-------
                                  Tiered Testing
                    Issues for Freshwater and Marine Sediments
                          Wednesday, September 16, 1992


8:30         Introduction and Description of EPA Sediment Strategy

             Elizabeth Southerland, Workshop Moderator, U.S. EPA Office of Science and
             Technology

9:00         Discussion of EPA Program Office Sediment Evaluation Needs

             Tom Armitage, U.S. EPA Office of Science and Technology

9:30         Discussion of EPA Regional Sediment Evaluation Needs

             William Peltier, Environmental Services Division, U.S. EPA Region 4

10:00        Break
                                            i   -
10:15        Tiered Sediment Testing Conceptual Overview

             Elizabeth Southerland, U.S. EPA Office of Science and Technology
                                                 '                                i
             Tom Armitage, U.S. EPA Office of Science and Technology

10:45   ,     Summary of ASTM  Activities on Freshwater and Marine  Sediment Test
             Methods

             Chris Ingersoll, U.S. Fish and Wildlife Service National Fisheries Contaminant
             Research Center

11:15        Discussion of Approaches for Test Standardization: Historical Perspective and
             Present Guidance

             Jim Lazorchak, U.S. EPA  Environmental Monitoring and Systems Laboratory,
             Cincinnati

             Bill Telliard, U.S. EPA Office of Science and Technology
                                       475

-------
12:15        Lunch


1:15         Discussion of Desirable and Necessary Attributes for Freshwater Sediment
             Toxicity Tests, and the Use of Hvalella  azteca and Chironomns teritans in
             Freshwater Sediments

             Allen Burton, Wright State University

             John Giesy, Michigan State University, Department of Fisheries

             Chris Ingersoll, U.S. Fish & Wildlife Service National Fisheries Contaminant
             Research Center

2:00         Discussion of Desirable and Necessary Attributes for Marine and Estuarine
             Sediment Toxicity Tests, and the Use of Ampelisca abdita. Rhepoxvnius
             abronius. Leptocheirus plumulosus. Eohaustorius  estuarius in Marine and
             Estuarine Sediments

             Rick Swartz, U.S. EPA Environmental Research Laboratory, Pacific Division

3:15         Break

3:30         Discussion of Desirable and Necessary Attributes for Freshwater, Marine,
             and Estuarine Bioaccumulation Test, and the Use of Neries and Macoma in
             Marine and Estuarine Sediments                                 .

             Peter Landrum, NOAA  Great Lakes Environmental Research Laboratory

             Henry Lee, U.S. EPA Environmental Research Laboratory, Pacific Division

4:45         Discussion of the Use of Lumbriculus variegatus hi Freshwater Sediments

             Gary Ankley, U.S. EPA Environmental Research Laboratory, Duluth

5:00         Identification of Long Term Needs for Assessing Sediments

             Gary Ankley, U.S. EPA Environmental Research Laboratory, Duluth

             Norm Rubinstein, U.S. EPA Environmental Research Laboratory, Narragansett

5:30         Adjourn
                                       476

-------
               Break-Out Workgroup for Freshwater Sediment Issues

                             Thursday, September 17




8:00 am     Introduction and Overview of Day

                                            Moderator/Discussion Leader: Gary Ankley
                   General description of survey/summary of methods
                   Number of labs responding, who responded on which organisms, et
                   cetera.

                   Discussion of issues for each method
8:30 am      Development of a Standard Protocol for Testing Hvalella azteca

                                            Discussion Leader: Teresa Norberg-King
                                          ,  Rapporteur:  Chris Ingersoll


             •     Summary of culture methods used as reported in questionnaires
                                                                     (-10 mm)

             •     Summary of test methods used as reported in questionnaires
                                                                     (~ 10 min)

             •     Proposed key issues for discussion



10:15 am    Break


10:30 am    Continuation of Discussion of Development of a Standard Protocol for Testing
             Hvalella azteca
                                       477

-------
11:30 am    Development of a Standard Protocol for Chironomus tentans

                                                 Discussion Leader:  Bob Hoke
                                                 Rapporteur: Jody Kubitz
             •      Summary of culture methods used as reported in questionnaires
                                                                    (-lOmin)

             •      Summary of test methods used as reported in questionnaires
                                                                    (~10min)

             *      Proposed key issues for discussion


12:15 pm     Lunch


1:30 pm      Continuation of Chironomus tentans


2:30 pm      Development of a Standard Protocol for Lumbriculus variegatus
                                                  t
                                                 Discussion Leader: Peter Lahdrum
                                                 Rapporteur: Allen Burton


             •      Summary of culture methods used as reported La questionnaires
                                                                    (~10min)

             •      Summary of test methods used as reported in questionnaires
                                                                    (~10min)

             •      Proposed key issues for discussion


3:30 pm      Break


3:45 pm      Continuation of Lumbriculus variegatus


5:00 pm      General Adjournment
5:30 pm     Meeting of Discussion Leaders,  Moderators, and Rapporteurs  to  Write
            Conclusions
                                      478

-------
          Break-Out Workgroup for Marine and Estuarine Sediment Issues
                            Thursday, September 17
8:00 am     Overview of Day
            •     Discussion of Issues for Each Method
            Norm Rubinstein, U.S. EPA Environmental Research Laboratory, Narragansett
8:30 am     Development of a Standard Acute Amphipod Protocol
            •     Summary of methods used
            •     Applications of the test
            •     Minimum requirements for the method
            •     Technical issues
            Rick Swartz, U.S. EPA Environmental Research Laboratory, 'Pacific Division
10:15 am    Break                                                    .
10:30 am    Development of a Standard Chronic Amphipod Protocol
            •     Summary of methods used
            •     Applications of the test
            •     Minimum requirements for the method
            John Scott, SAIC
            Ted DeWitJ, AScI
12:15 pm    Lunch
1:30 pm    Continuation of Chronic Protocol Discussion
                                      479

-------
2:30 pm     Development of a Standard Bioaccumulation Protocol



            •     Summary of methods used



            •     Applications of the test



            •     Minimum requirements for the method




            •     Technical issues



            Henry Lee, U.S. Environmental Research Laboratory, Pacific Division




3:30 pm     Break



3:45 pm     Continuation of Bioaccumulation Protocol Discussion




5:00 pm     Development of Other Test Methods and Standard Protocols




5:30 pm     Adjourn
                                      480

-------
                                  Tiered Testing
                    Issues for Freshwater and Marine Sediments
                              Friday, September 18
9:00 am     Overview of Day
                                           "                      > •
            Elizabeth Southerland, Workshop Moderator, U.S. EPA Office of Science and
            Technology

9:15 am     Report of Issues Covered in Marine Methods Session and  Discussion of
            Conclusions/Next Steps
                         ;                        ' •
            Rick Swartz, U.S. EPA Environmental Research Laboratory, Pacific Division

10:15 am    Report of Issues Covered in Freshwater Methods Session and Discussion of
            Conclusions/Next Steps

            Gary Ankley, U.S. EPA Environmental Research Laboratory, Duluth
                                                                [
11:15 am    Workshop Summary and Wrap-up

            Elizabeth Southerland, Workshop  Moderator, U.S. EPA Office  of Science and
            Technology

11:45 am    Adjourn
                                        481

-------

-------
      APPENDIX B:

SEDIMENT TOXICITY TESTS
 UNDER DEVELOPMENT BY
 ENVIRONMENT CANADA
           483

-------

-------
DEVELOPMENT OF A 10-DAY MARINE/ESTUARINE AMPHIPOD ASSAY
FOR SEDIMENT TOXICITY IN SUPPORT OF THE CANADIAN OCEAN
PUMPING PROGRAM (CEPA, PART VI).
D.J. McLeay, S.C.  Yee, K.G. Doe and L.M. Porebski, McLeay Associates Limited, West
Vancouver, British Columbia, Environment Canada, and Aquatic Toxicity Laboratory, North
Vancouver, British Columbia, and Environment Canada, Laboratory Division, Dartmouth, Nova
Scotia, and Environment Canada, Office of Waste Management, Environment Canada, Ottawa,
Ontario.

                                   ABSTRACT

       Beginning in  1988, Environment Canada commenced the development of a test method
for measuring the acute toxicity of sediment samples, using a number of marine or estuarine
sediment-burrowing  amphipods common to Canada's coastal waters.   The evolution of this
toxicity test included five series of inter-laboratory assessments using  various candidate test
organisms, sediment samples, and a reference toxicant.  These studies are reviewed briefly.
Additionally, the test method and its applications are summarized.             ,


                                 INTRODUCTION

       The Ocean Dumping Control Act has been consolidated into Part VI of the Canadian
Environmental Protection Act (CEPA). The Act requires valid ocean dumping permits before
dumping of any substance at sea is allowed. Biological testing and assessment can be an integral
component of this permit process (Sergy, 1988; Anthony, 1991; Porebski, 1991).  In order for
Environment Canada to perform its regulatory responsibilities associated with this Act, biological
screening tests using marine or estuarine organisms may be required to determine if material is
suitable for unconfmed open-water disposal, and to perform environmental-effects monitoring
at dump  sites.   Interim contaminant testing guidelines for ocean disposal,  associated with
Environment Canada's Ocean Dumping Control Program, specify several toxicity tests for use
in screening materials.  The 10-day assay  for sediment toxicity, using one  or more species of
estuarine or marine amphipods common to Canada's coastal waters, is included on the lists of
biological screening tests (Environment Canada, 1990a, 1990b).

       In consideration of the above, Environment  Canada's Atlantic and Pacific & Yukon
regional laboratories commenced inter-laboratory studies in late 1988, using a number of marine
or estuarine sediment-burrowing amphipods and various samples of control, reference or test
 (contaminated) sediments.  The objectives of this testing program were twofold: (1) to study
 several candidate species of amphipods, selecting those suitable for use in  acute lethality tests
 with samples of sediment or other test material; and (2) to evaluate conditions, procedures and
 biological endpoints appropriate for use in a standard biological test method to be developed to
 meet Environment Canada's testing requirements in this respect.  The past status of this and
 other biological test methods under development on behalf of Environment Canada has been
 reported (McLeay etal., 1991a)..   ,

                                        485 -•-.'•'

-------
       Following is a brief summary of the inter-laboratory studies with marine or estuarine
 amphipods, performed to date by Environment Canada's Pacific & Yukon and Atlantic regional
 laboratories.1   Also presented  is a  list  of those species of infaunal amphipods  presently
 recommended for use in Environment Canada's draft biological test method "Acute Test for
 Sediment Toxicity Using Marine or  Estuarine Amphipods"  (Environment Canada,  1991).
 Tentative checklists of recommended conditions and procedures for holding and acclimating
 amphipods, and for testing them in 10-day static assays, are provided.   Finally, some of the
 applications of this biological test method are indicated.                      •
      LABORATORY EVALUATION AND DEVELOPMENT OF TEST METHOD

       In  1988, Environment Canada's  Atlantic and  Pacific-& Yukon regional laboratories
undertook a  preliminary (Phase-I)  evaluation  of the  10-day  sediment assay,  using only
Rhepoxynius  abronius  (McLeay et al., 1989).  The effects of holding amphipods in control
sediment for periods of up to 81 days, on their 10-day survival and subsequent reburial rates
under test  conditions (control sediment only), were investigated.  Their acute tolerance (96-h
LC50/EC50)  to the reference  toxicant  cadmium chloride (seawater-only  exposures) was
monitored  during the prolonged  holding period.  Although 109-day survival and reburial rates
were acceptable (> 90%) in all trials, the reference toxicant tests indicated a declined tolerance
of these organisms with extended > 13 days) periods of holding in the laboratory.

       The second  (Phase-IE) inter-laboratory study  (McLeay  et al., 199Ib)  measured and
compared  the 10-day survival, emergence and reburial rates for a population of Rhepoxynius
abronius exposed to control,2 reference,3 or test* sediment.  At each laboratory, survival and
reburial  rates in control sediment were high (>90%).   No consistent differences in these
endpoints were caused by exposure to reference or test sediments, indicating that neither were
highly toxic.  However, increased rates of emergence from the test sediment were  observed.
   'Basic test conditions and procedures used for the 10-day static assays were according to Swartz et al. (1985)
and ASTM (1990).

   2Control sediment is clean sediment taken from the site where the test organisms were collected, and intended
for use in the 10-day test*with amphipods. This sediment must contain 'no test material.  It is used to determine the
absence of measurable toxicity due to basic test conditions (e.g., temperature, health or handling of test organisms).

   3Reference sediment is a field-collected sample of sediment, taken from a site thought to be relatively free of
contaminants (i.e., "clean" sediment), and intended for use in the 10-day test with amphipods.  It is often collected
from a site within the general  vicinity of a test sediment, and is frequently selected for biological testing because
of its geochemical similarity (e.g., particle size, compactness, total organic content) to the test sediment(s).

   ^Test sediment is a field-collected sample of solid-phase sediment, taken from a site thought to be contaminated
with one or more chemicals, and intended for use in the 10-day test with amphipods. In this study, the sample of
test sediment was collected from the vicinity of a B.C. coastal pulp mill discharging bleached kraft mill effluent.

                                           486

-------
       A Phase-m study was performed by each laboratory using both Rhepoxynius abronius
and Corophium volutator as test organisms (McLeay et al., 1991b).  For each species,  ten-day
survival and subsequent reburial rates were determined for three clean5 (control or reference)
sediments and three test sediments.  The clean sediments varied appreciably in grain size, with
silt/clay contents of 1%, 82% or 99%.   Tests with R, abronius showed highest survival rates
(96% and 100%) for control sediment, and lowest survival rates (43% and 78% for the reference
sediment containing 99% fines  (silt and clay).  R, abronius  survival rates  for the three test
sediments ranged from 72 to 93%.   Unlike these findings, survival rates for Corophium
volutator were high (93 % and 97%) in the extremely fine-grained reference sediment. For both
species,  reburial and/or survival data6 showed no consistent  response for any of  the test
sediments examined, although emergence rates indicated an avoidance response to one of the test
sediments.

       A fourth inter-laboratory assessment,7 using six species of marine or  estuarine  mfaunal
amphipods8 common to Canada's coastal waters, was undertaken during late 1990 and early
1991 (Paine and McPherson, 1991a).  The objectives of this (Phase-IV) study  were to determine
and compare the relative sensitivity of  each of the six test species to four test sediments, two
reference sediments (fine-grained and  coarse-grained),  and control sediments (one for each
species)  For the four test sediments examined, each of the six species of amphipods used in
these assays distinguished  the same two sediments as clearly toxic, and the remaining two as
marginally or not toxic. Percentage survival at 10 days was the most useful biological endpoint;
little if any additional information was  obtained using the other two endpoints (% emergence,
 %  of survivors that did not rebury in control sediment within 1 h following test completion).
R  washinetonianus and K abronius were most sensitive to the test sediments; C volutator and
E  estuarius were least sensitive.  Two of the six species studied (R  washingtomanus and A,
vh-giniana)  showed unacceptably low (< 90%)  10-day survival rates  in  control sediment;
Depending  on  species,  grain-size effects  were  minimal or not evident.   This  study also
demonstrated that, if care is taken, amphipods can be shipped across  the country without
influencing the test results.       .                      ..  ...  '

        Additional studies were performed by Environment Canada's Atlantic regional laboratory
 in  early 1991 to assess the laboratory hardiness and worth of Amphiporeia virginiana as a test
 organism (Doe, 1991).  Two populations of field-collected specimens were tested  for 10-day
 survival rates in control sediment only.  Animals were acclimated and tested at temperatures of
 10 or 15 C (Series 1); and at 5, 10 or 15 C (Series 2). Results from these studies showed_that
 acceptable  (>90%)  10-day survival rates  could be obtained at 5 or 10 C? but not  at 15 C.
 Seasonally-cold (2 to 4 C) seawater temperatures at collection  sites likely contributed to these
 findings.                                                        ,
    *Clean sediment is sediment (e.g., control or reference sediment) that does not contain concentrations of
 contaminants which cause discernible distress to the test organisms or reduce their survival in 10-day assays.

    'Since a volutator do not rebury readily in control sediment within 1 hour, this biological endpoint cannot be
 determined for this species.                               ~                                .

    Participating laboratories included Environment Canada's Atlantic and Pacific & Yukon regional laboratories,
 and EVS Consultants Ltd.
     Test organisms were Rhepoxvnius abronius.  Fo^EbaM xiximeus,  Eohaustorius estuarius, Eohaustorius
  washingtonianus. Corophium volutator. and Ampfiiporeia virginiana.
                                            487

-------
        A fifth inter-laboratory9 appraisal of the 10-day test for sediment toxicity, using multiple
 species of  marine or  estuarine  amphipods,  was  conducted during  June 1991 (Paine and
 McPherson 199Ib).  For this study, seven species of infaunal amphipods10 were collected and
 examined for their laboratory adaptability and sensitivity to each of the three test sediments.
 Biological endpoints  (%  survival and %  emergence at  10 days; % reburial of survivors in
 control sediment at test end) were determined  for each species in these sediments as well as in
 a fine-grained (79% silt-clay) reference sediment and respective control sediments. Assays with
 Amphiporeia virginiana were performed at both  10 and 15 C; all other species were tested at
 15  C only.  Unlike  the previous (Phase-IV) inter-laboratory study, none of the three test
 sediments used in this study were highly toxic to any of the species of amphipods examined.
 Once again, ten-day  survival rates  in  control sediment were unacceptably low (,90%)  for
 Amphiporeia virginiana (both temperatures) and Eohaustorius washingtonianus.  but acceptable
 for all other species  studied including Leptocheirus pinguis.  As in the Phase-IV study,  %
 survival  at  10 days  was the most  useful biological endpoint,  with  little if any additional
 information provided  by the  secondary endpoints (% emergence, % of survivors not reburying
 in control sediment at test end).


                           RECOMMENDED TEST SPECIES

        Recent attempts  have been made to identify suitable Canadian collection sites," and to
 collect the following species  of amphipods from Canadian coastal waters, in order to evaluate
 their worth as candidate test  organisms in  10-day sediment assays:


 Pacific Coast                                Atlantic Coast

 Monoculodes spinipes                        Rhepoxynius hudsoni
 Grandifoxus grandis                          Phoxocephalus holbolli
                                              Ampelisca abdita                        .
                                              Ampelisca vadorum
                                              Amphiporeia lawrenciana
                                              Pontoporeia femorata

The absence or relative scarcity of these species at the Canadian  collection sites investigated,
prevented their inclusion in the present test program.
   'Environment Canada's Atlantic and pacific & Yukon regional laboratories.

   10Test organisms included Rhepoxvnius abronius. Foxiphalus xiximeus. Eohaustorius estuarius. Eohaustorius
washingtonianus. Corophium volutator. Amphiporeia virginiana. and Leptocheirus pinguis.

   "Specimen collections held by the National Museum of Natural Sciences (Ottawa, Ontario) were examined by
Dr. E.L. Bousfield, together with historical records available to him (e.g., Bousfield, 1990a, b; Bousfield, 1991;
Bousfield and  Laubitz, 1972).  Based on these and other distributional and life-history information,  initial
recommendations were made for prospective test specimens.
                                                                   J

                                           488

-------
       Based on the results  of the laboratory studies mentioned  previously (see section
 "Laboratory Evaluation and Development of Test Method"), as well as those for other 10-day
 assays performed with the candidate species of irifaunal amphipods under consideration by
.Environment Canada, the following species of marine or estuarine amphipods are presently
 recommended for use in 10-day static sediment assays (Environment Canada, 1991):
  Recommended Pacific Species

  Rhepoxynius abronius
  Foxiphalus xiximeus
  Eohaustorius estuarius  ,
Recommended Atlantic Species

Corophium volutator
Leptocheirus pinguis
       Additional studies, with Ajnphiporda virgihiana and Eohaustorius washingtonianus are
 required to demonstrate that satisfactory survival rates in control sediment can be achieved for
 these species, before they can be recommended by Environment Canada as standard  test
 organisms.                                          ,


                RECOMMENDED CONDITIONS AND PROCEDURES
                 FOR ACCLIMATING AND TESTING AMPHIPODS
 \       _     • .                                 •                ',
       Environment Canada's draft biological test method (Environment Canada, 1991) provides
 details regarding conditions and procedures for holding  and acclimating amphipods to be used
 in 10-day assays, as well as those necessary to perform the test in a standardized manner. The
 test methods of Swartz et al. (1985)  and  ASTM (1990) formed the basis for Environment
 Canada's (1991) acute test for sediment toxicity using marine or estuarine amphipods common
 to Canadian coastal waters.  Acclimation and test procedures  recommended in Environment
 Canada (1991) are  reproduced here (see Tables 1 and 2). Since this biological test method is
 not yet finalized and  approved by Environment Canada, these procedures and conditions are
 subject to change.
                                          489

-------
Table 1      Checklist  of Recommended Conditions  and  Procedures for  Holding  and
             Acclimating Amphipods12
 Source of amphipods:

 Life stage:

 Sorting:
 Holding sediment:
 Holding seawater:
 Acclimation conditions:
 Lighting:



 Feeding:

 Duration of acclimation:

 Health criteria:
 Collected subtidally or intertidally
 from clean sediment

juveniles or young adults, 3-10 mm
 length (depending on species)

 Sieve through 1.0-mm screen to
 confirm species and select
 appropriate size;  use seawater within
 2 C and 2 ppt salinity of the^
 seawater in transport container

 Control sediment, 2-4 cm in depth,
previously sieved through 0.5-mm
 mesh

Reconstituted or clean natural
 seawater

Salinity of seawater same as that for
overlying seawater in test;,
temperature normally 15 +  2 C;
dissolved  oxygen > 90% of air
.saturation; temperature, salinity, and
dissolved  oxygen measured and
recorded daily

Constant overhead illumination, >
 100 lux at surface of sediment in
holding/acclimation container(s)

None

3 to 10 days

Select amphipods able to bury
quickly in control sediment; remove
inactive amphipods  that have
emerged from sediment or do not
bury; discard population if  ^5 %
dead or emerged and inactive during
48-h period preceding test
   12From Environment Canada's (1991) draft biological test method - subject to change.

                                 "490

-------
    APPENDIX C:



FRESHWATER SURVEYS
         491

-------

-------
                               Culturing Questionnaire

             Species:    ,  	—	—	:——
             Laboratory:   —	'•	:	——
             Contact:      	•	

1.      Is the culture intermittent or continuous? If continuous, how long have animals been in
       culture at your facility?

2.      What was original source and approximate date you started the culture?

3.      Have the animals been taxonomically identified?  If so, when and by whom?

4.      what records on culture animals are maintained?

                                                circle one        Frequency
       parental survival                          ves    no
       age of brood animals                     yes  t  no	.	—
       temperature                             ves    no	—
       dissolved oxygen          ......      yes    no   	.	:—
       pH                                     yes    no	
       quality/age of foods                      yes    no   ,	—.	—
       frequency of new culture chambers        yes    no	:	=—
 5.     What is source of the water used for culturing?  List all used.

 6.     What are the characteristics of each water?

       water         hardness      alkalinity           BS "  .   -   conductivity

 7.     Have you used any reconstituted waters for culturing? If so, are they successful? If no,
       what problems did you experience?

 8     What  foods have you tried to culture the animals?  What is your choice of food for
       routine culture?               '

 9.     How long has this regime been used?

 10.   Do  you culture under  controlled  temperature  (ฑ2ฐC) and lighting?  If so, specify
       conditions.

 11.   What substrates have you used for culturing? Please list types/quantities evaluated and
       current choice.
                                            493

-------
12.    Do you conduct any reference toxicant tests with this organism?

       A.     If so, with what toxicant? What duration and was test conducted with or without
              sediment? If with sediment, specify source/type.

       B.     Have results been reproducible?  Have any control charts been established?

13.    Do  you   feel  reference  toxicant   tests  are   relevant   for   monitoring   the
       adequacy/performance of the culture organisms?

14.    Provide any additional information or comments that are pertinent to this species below:
                                        494

-------
                                Testing Questionnaire                         .
        1                      -,                      >            '                         • •
             Species:      —.—	—	!	
             Laboratory:  ^_—	—-———          ';        ' ';
             Contact:	—•	;—

1.     Describe test (toxicity, bioaccumulation).

2      Are the test organisms  cultured at your laboratory?  If yes, provide description in
   '    Attachment 1.  If no, please provide details of how test animals are obtained.

3.     What is the "routine" test performed with this organisms at your laboratory?  Select the
       appropriate time frame.                             •   •

       .	 4-d    	7-d   •   •'•  10-d	 14-d	 21-d
       	other, specify		—-      •

4     Do you use known age or size or unknown age or size of organisms to initiate tests?
       What is known age or size?   Specify age or size.  How do you obtain  these known
       age/size organisms? .

5.     Is it important that animals are a minimum age (or size) for sediment:water exposures
       to ensure recovery (of organisms or sediments)?

6.     Have you specified a certain test design of sediment:water?   If so, what and why?    -

7.     What is the renewal frequency of the overlying water for the  duration of the  test?  What
       procedure is used to renew the water?

 8.     Do you feed during the test?  Is  it the same rate and frequency as in the culture?  If not
       specify what and why.                                     :

 9.     What statistical analyses are performed on the data?  Cite specific procedures and types
       of statistical analysis.

  10.   What are the test endpoints?  How do you express the effect?

  11    Have  you conducted any reference toxicant tests in your  laboratory sedimentrwater
       exposures? If yes, please identify chemicals and  explain general trend of  test results.
       Manuscripts can be attached.
                               " •                           V             •
  12.   What water quality characteristics of the overlying water are measured during the test?
       How often are these parameters measured?
                                            495

-------
13.    Provide any additional information or comments that are pertinent to this species below:
                        PLEASE PROVIDE DATA IN SPACE BELOW
                     CULTURE CONDITIONS FOR HYALELLA AZTECA
CULTURE CONDITION
  1.   Culture type (static or renewal)
      (specify rate)
  2.   Temperature:
  3.   Light quality:
  4.   Light intensity:
  5.   Photoperiod:
  6.   Culture chamber size:
  7.   Culture water volume:
  8.   Frequency of starting new culture
  9.   Renewal  of culture water:
10.   Removal of offspring (frequency):
11.   Age of restart organisms:
12.  No. organisms/culture chamber:
13.  No. of culture tanks:
14.  Feeding regime:
15.  Substrate used:
16.  Chamber cleaning:
17.  Aeration:
18.  Culture water:
CONDITION USED BY LABORATORY
                                      496

-------
                      PLEASE PROVIDE DATA IN SPACE BELOW
                  CULTURE CONDITIONS FOR CHIRONOMUS TENTANS
        CONDITION                         CONDITION USED BY LABORATORY

 1.   Culture type (static or renewal)       .'•".'
     (specify rate)                          '_	.	         •   -
 2.   Temperature:                      , '.  ——•.	—	
 3. •  Light quality:                          	—	—	
 4.   Light intensity:                        ——	:	-^	—
 5.   Photoperiod:                                     /	-	
 6.   Culture chamber size:                  	——	
 7.   Culture water volume:                 ——	—	
 8.   Frequency of  starting new culture	—	;	
 9.   Renewal of culture water:              	   /       	
10.   Removal of offspring (frequency):       	.	——-—..	,	
11.   Age of restart organisms:               ;	—	;—.
12.   No. organisms/culture chamber:         	——:	——	
13.   No. of culture tanks:  .                 	—	-—-—r^	
14.   Feeding regime:                       —	;—•	——	—
15.   Substrate used:                        	——	:	—	
16.   Chamber cleaning:                     —-,	—	•	—-
17.   Aeration:                             -—:—;	:	
18.   Culture water:	—=	~^-
                                     497

-------
                       PLEASE PROVIDE DATA IN SPACE BELOW
                 CULTURE CONDITIONS FOR LUMBRICULUS VARIEGATUS
CULTURE CONDITION
  1.  Culture type (static or renewal)
     (specify rate)
  2.  Temperature:
  3.  Light quality:
  4.  Light intensity:
  5.  Photoperiod:
  6.  Culture chamber size:
  7.  Culture water volume:
  8.  Frequency of starting new culture
  9.  Renewal of culture water:
10.  Removal of offspring (frequency):
11.  Age of restart organisms:
12.  No. organisms/culture chamber:
13.  No. of culture tanks:
14.  Feeding regime:
15.  Substrate used:
16.  Chamber cleaning:
17.  Aeration:
18.  Culture water:
CONDITION USED BY LABORATORY
                                      498

-------
            PLEASE PROVIDE INFORMATION IN SPACES BELOW IF APPROPRIATE
             CONDITIONS FOR HYALELLA AZTECA SEDIMENT TOXICITY .TESTS
TEST CONDITION             '                 CONDITION USED BY LABORATORY
  1.  Test type (static or renewal)
     (specify rate)                           	;	
  2.  Test duration:                          -—	—;—	—
  3.  Temperature:	•	
  4.  Light quality:	———	•-
  5.  Light intensity:                 .                   ,.————	——-
  6.  Photoperiod:	:	:	-^——
  7.  Test chamber size:                     ——.	:	
  8.  Test sediment volume:                  .	—;	:	•	•	
  9.  Overlying water volume: chamber:       	•.	:	—-
 10.  Age of test organisms:                  .	.	——
 11.  Size of test organisms:          .*    _   —	=	—=	:—
 12.  No. organisms/test chamber:            —.	;	•	
 13.   No. replicate test chambers/sediment:    _	——
 14.   No. organisms/sediment:	—;	—	
 15.   Feeding regime:  .                     	;	-.	:—:	
 16.   Test chamber cleaning:     .            	—	.  •   .  	
 17.   Aeration:                 .             ——	;—
 18.   Overlying water quality characteristics:   _—;	——;	
 19.   Test acceptability criterion:,             	—	
      (1) minimum control survival           ——^	———	
      (2) length or weight minimum criteria:   .	-—,	—
 20.   Eridpoint(s)                        .   —	—	—
NOTE: If an item listed does not seem pertinent, please explain why.
                                       499

-------
              PLEASE PROVIDE INFORMATION IN SPACES BELOW IF APPROPRIATE
             CONDITIONS FOR CHIRONOMUS TENTANS SEDIMENT TOXICITY TESTS
  TEST CONDITION                             CONDITION USED BY LABORATORY
   1.  Test type (static or renewal)
       (specify rate)
   2.  Test duration:
   3.  Temperature:
   4.  Light quality:
   5.  Light intensity:
   6.  Photoperiod:
   7.  Test chamber size:
   8.  Test sediment volume:
   9.  Overlying water volume: chamber:
  10.  Age of test organisms:
  11.  Size of test organisms:
  12.  No. organisms/test chamber:
  13.  No. replicate test chambers/sediment:
  14.  No. organisms/sediment:
  15.  Feeding regime:    '
  16.  Test chamber cleaning:
  17.  Aeration:
  18.  Overlying water quality characteristics:
  19.  Test acceptability criterion:
       (1) minimum control survival
       (2) length or weight minimum criteria:
  20.  Endpoint(s)
NOTE: If an item listed does'not seem pertinent, please explain why.
                                        500

-------
            PLEASE PROVIDE INFORMATION IN SPACES BELOW IF APPROPRIATE
         CONDITIONS FOR LUMBRICULUS VARIEGATUS SEDIMENT TOXIC1TY TESTS
TEST CONDITION                              CONDITION USED BY LABORATORY
 •1.  Test type (static or renewal)
     (specify rate)                   ,        ;	_	
  2.  Test duration:                          ——	—	
  3.  Temperature:                           	:—	;	——
  4.  Light quality:   ,                       —	———:	;—
  5.  Light intensity:                         ——	—
  6.  Photoperiod:                           	•	—	
  7.  Test chamber size:                     ——	—	
  8.  Test sediment volume:	—	—
  9.   Overlying water volume: chamber:       _	:	—	
 10.   Age of test organisms:                   '    	—-———	
 11.   Size of test organisms:                  	'•	—	—
 12.   No. organisms/test chamber:             	——;	••	:—
 13.   No. replicate test chambers/sediment:,    _	:	—	——
 14.   No. organisms/sediment:                	;	:—	—-.	
 15.   Feeding regime:	———
 16.   Test chamber cleaning:                 	;	:	—	
 17.,  Aeration:                              —	:	:	:	•
 18.   Overlying water quality characteristics:	———:
 19.   Test acceptability  criterion:              	_—	:	•—
      (1) minimum control survival            	:—_	;	
      (2) length or weight minimum criteria:   ,—	=:—.	.	—-—
 20.   Endpoint(s)                           —	*	
NOTE: If,an item listed does not seem pertinent, please explain why.
                                        501

-------
                            General Culturing Issues

                                   Substrate
                                    Density
                                     Water
                             Flow-through vs. Static
                                 Feeds/Feeding
                         Genetic drift/stream differences
                               Known age systems
                               Nuisance organisms
                              . Light/photpperiod
                                  Temperature
                 QA/QC (e.g., reproduction, reference toxicants)
                             General Testing Issues

                             Test lengths/endpoints
                                 Organism age
             Water Renewal (volumes, frequency, method of renewal)
                   Physical Test System (sediment volume, etc.)
                Test condition and design (chambers, lighting, etc.)
Interpretation of sediment variables (e.g., organic carbon, particle size) on test results
                             Feeds/feeding regimes
                           QA/QC for acceptable tests
                                   502

-------
Table 1. Survey respondents for sediment test organisms.
Laboratory 	 ' 	 	
Department of Fisheries & Oceans
Environment Canada
EPA-Duluth
EPA Region 1
EPA Region 8
EPA Newtown
EVS Consultants
Maryland Department of Environment
Miami University
Michigan State •
NFCRC-Athens ,
NFCRC-Columbia
NO A A- Ann Arbor
Old Dominion
State of Washington
University, of Mississippi
University of Wisconsin-Superior
Wright State
H. azteca
X
X
X
X
X
X
X
X
' ' x
X
X
X
X
X
X

X
X
C. tentans/
C. riparius L. variegatus*
' '
X
X X
X

X
X~


X X
X
x x
X X


x
X X
X
  a     Two responses were on two different species.
  Note: EPA region 6, Dallas, Texas, and the FDA-Center for Veterinary Medicine,
        Maryland, responded to survey but did not have organisms in culture.
                                         503

-------

-------
      APPENDIX P:

     BIBLIOGRAPHIES
          FOR
FRESHWATER TEST SPECIES
            505

-------

-------
Group: C:\REF\CfflRSCHl.GRP
Temporary group for searching
 Sorted by: Authors, Year, Title
 Using Format LMNOL OCEANOGR
 Current Search: term=chir
 Last Search run on 9 Sep 1992, at 11:18
 Last modified on 9 Sep 1992, at'12:53

Listed with Format LIMNOL OCEANOGR

Listing Created 9 Sep 1992, at 12:55
ADAMS W.  J., R.  A.  K3MERLE, AND R.  G.  MOSHER.  1985.  Aquatic Safety  Assessment  of
        Chemicals Sorbed to Sediments, p. 429-453. In R. D. Cardweli, R. Purdy, and R. C. Bahner [eds.],
        Aquatic Toxicology and Hazard Assessment, ASTM STP 854, American Society of Testing and
        Materials.

ADAMS  W.   J.,   P.   S.  ZEGENFUSS,  W.  J.  RENAUDETTE,  AND  R.  G.  MOSHER.  1986.
        Comparison of laboratory and field methods for testing 'the'toxicity  of chemicals  sorbed to
        sediments,  p.  494-513.  In  T. M. Poston,  and R. Purdy  [eds.],  Aquatic Toxicology and
        Environmental Fate. ASTM STP 921, American Society for Testing and Materials.

ANDERSON  R.  L.,  AND  P.   SHUBAT.   1983.  Insecticide  effects   on  normal,  development  and
        hatch  of embryos  of Paratanytarsus  parthenogeneticus (DipteraChironomidae). Great Lakes
        Entomol. 16: 177-181.                               ,                                 -

ANDERSON  R. L.,  C.  T.  WALBRIDGE,  AND  J. 1 T.  FIANDT.  1980.  Survival  and  growth  of
        Tanvtarsus dissimilis exposed to copper, cadmium, zinc and lead. Arch. Environ. Contain. Toxicol.
      '. 9: 329-335.                                        "                   '

AROKLASAMY F.  J.,  AND  C.  J.  DODGE.  1990.  Anaerobic  microbial   remobilization of  toxic
        metals coprecipitated with iron oxide. Environ. ScL Technol. 24: 373-378.

BARKER   D.  M.,  AND  J.  WILHM.  1982.   Oxygen   consumption  rates  in  sediments  and  of
        Chironomus riparius (Meigen) and Cbaoborus punctipennis (Say) in an artificially destratified lake.
        1.1:607-613.                                                         .

 BARTON   D.  R.  1989a.  Some  problems  affecting  the   assessment  of Great   Lakes water  quality
        using benthic invertebrates. J. Great Lakes Res. 15: 611-622.
                                                                      s"
 —.  1989b.  Some  problems  affecting  the  assessment  of  great  lakes  water quality using  benthic
        invertebrates. J. Great Lakes Res. 15: 611-622.                      .

'BERG M.   B.,  AND  R.  A.   HELLENTHAL.  Data  variability  and   the  use  of  chironomids  in
        environmental studies:  The standard error of the midge, pp.1-8 in W.S. Davis (ed). Proceedings
        of the 1990 Midwest Pollution Control Biologists Meeting. U.S. Environmental Protection Agency,
        Region V, Environmental Sciences Division, Chicago, IL.  EPA-905-9-90/005.

 BIEVER  K;  D.  1965,  A  rearing  technique   for  the  colonization  of chironomid midges. Ann.
        Entomol. Soc. Amer. 58: 135-136.                           '     -
                                                    ft              '                           •-
                                                                '                                '
                                                   567

-------
BRENNAN  A.,  AND  A.  L  MCLACHLAN.  1979. Tubes  and  tube-building in  a lou'c  chironomid
        (Diptera) community. Hydrobiologia 67: 173-178.

BRINKHURST  R.   O.   1989.  A   phylogenetic  analysis   of   the   Lumbriculidae   (Annelida,
        Oligochaeta). Can. J. Zool. 67: 2731-2739.

BUHL  K.  L,  AND  N. L.  FAERBER. 1989.  Acute  toxicity  of  selected  herbicides  and surfactants
        to larvae of the midge Chironomus riparius. Arch. Environ. Contain. ToxicoL 18: 530-536.

BURTON  JR.  G.   A.  1991.   Assessing  the   toxicity   of   freshwater  sediments.  Environ.  Toxicol.
        Chem. 10: 1585-1627.

BUTLER  M.  G.,  AND D. H. ANDERSON.  1990.  Cohort , structure,  biomass,  and  production  of
        a merovoltine Chironomus population in a Wisconsin bog lake. J. N. Am. Benthol. Soc. 9: 180-
        192.

CAIRNS M.  A., A. V.  NEBEKER,  J. H.  GAKSTATTER, AND  W.  L. GRIFFIS.  1984.  Toxicity
        of copper-spiked sediments to freshwater invertebrates. Environ. Toxicol. Chem. 3: 435-445.

CARR  R.  S., J.  W.  WILLIAMS,   AND  C.  T.  B.  ERAGATA.  1989. Development  and  evaluation
        of a novel marine sediment pore'water toxicity test with the polychaete Dinophilus gyrociliatus.
        Environ. Toxicol. Chem. 8: 533-543. "

CURRY   L.   L.   A  Survey   of   the   Environmental  Requirements   for  the   Midge
        (DipterarTendipedidae). Biological Problems in Water Pollution-3rd Seminar.

DANKS  H.  V.  1971.  Overwintering  of  some  north  temperate  and   arctic  Chironomidae   n.
        Chironomid biology. Can. Entomol. 103: 1875-1921.

—.   1978.  Some effects of  photoperiod,  temperature,  and  food  on emergence  in  three  species  of
        Chironomidae (Diptera). Can. Entomol. 110: 289-300.

DAVIES B.  R. 1976. The  dispersal of Chironomidae  larvae:  a  review.  J.  Ent  Soc.  Sth.  Afr.  39:
        39-62.

DERMOTT  R-  M.,  AND  C.   G.  PATERSON.  1974.  Determining  dry  weight  and  percentage  dry
        matter of chironomid larvae. Can. J. Zool. 52: 1243-1250.

DERR  S.  K.,  AND  M.   J. . ZABIK.   1972.   Biologically  active   compounds  in   the   aquatic
        environment: The uptake and distribution of [l,l-dichloro-2,2-bis(p-chlorophenyl) ethylehe], DDE
        by Chironomus tentans Fabricius (Diptera: Chironomidae). Trans. Amer. Fish. Soc. 2: 323-329.

—.   1974.  Bioactive  compounds hi  the  aquatic  environment:  studies  on  the  mode  of uptake  of
        DDE by the aquatic midge, Cfaironomus tentans (Diptera: Chironomidae). Arch. Environ. Contain.
        Toxicol. 2:  152-164.

DONALD  G.  L.,  AND C.  G. PATERSON.  1977.  Effect  of preservation  on  wet  weight  biomass
        of chironomid larvae. Hydrobiologia S3: 75-80.

EDWARDS  R.  W.,  H.  EGAN, M.  A.  LEARNER,  AND  P.  J.  MARIS.  1964.  The  control  of
        chironomid larvae in ponds, using TDE (ODD). J. Appl. Ecol. 1: 97-117.
                                                 508

-------
FAIRCHILD  W.,  D.  MUIR,  R/ CURRE,  AND  A.  YARECHEWSKI.   1992.  Emerging  Insects
        as a BiotiC Pathway for Movement of 2,3,7,8-Tetrachlorodibenzofuran from Lake Sediments.
        Environ. Toxicol. Chem. Ill 867-872.
HSHER  J.  B,,  AND  G.  MATISOFF.  1981.  High  resolution  vertical  profiles  of pH  in  recent
       sediments, Hydrobiologia 79: 277-284.

FISHER  S   W. 1991.  Changes  in the  Toxicity of three pesticides  as a function  of  environmental
       pH and temperature. Bull. Environ. Contain. Toxicol. 46: 197-202.

FISHER  S.  W.,  AND  R. W. WADLEIGH.  1986. Effects of pH on  the acute tozicity  and  uptake
        of 14*C pentachlorophenol in the midge; Cfaironomus riEanus. Ecotox. Environ. Safety 11: 1-8.

FOX  G   A  D   V  WESELOH,  T.  J.  KUBIAK,   AND - T.  C. ERDMAN.   1991.  Reproductive
      ' outcomes in colonial fish-eating birds:  a biomarker for developmental toxicants in Great Lakes
        food chains. I. Historical and ecotoxicological perspectives. J. Great Lakes Res. 17: (153-157.

FRANCO  P  J   K.  L  DANIELS,  •&  M.  CUSHMAN,   AND  G.  A.  KAZLOW.   1984.  Acute
        toxicity of a synthetic oil, aniline and phenol to laboratory and natural populations of chironomid
        (Diptera) larvae. Environ. PolluL (A)(i4: 321-331,

FRANK  C    1982.  Ecology,  production;  and  anaerobic metabolism  of  Chironomus  plumosus  L.
        larvae in a shallow lake LEcology and Production. Arch. Hydrobiol. 94: 460-491.

 FRY  D   M  AND  S  W  FISHER.  1990.  Effect of  sediment contact  and uptake mechanisms  on
        accumulation of three chlorinated hydrocarbons in the midge, CMronomus riEanus. Bull. Environ.
        Contam. Toxicol. 44: 790-797.   ;                                                        .

 FUKUHARA   H.,   AND  ,M.   SAKAMOTO.   1987.   Enhancement   of:  inorganic•   ซtaya   and
        phosphate release from lake sediment by tubificid worms and chironomid larvae. Oikos 48:312-
        320. .                   :"  '   '             '                                .

 FUKUHARA   H.,   AND   K.   YASUDA.   1989.   Ammonium   excretion   by   some   freshwater
        zoobenthos from a eutrophic lake. Hydrobiologia 173: 1-8.

 FULLER  R.  L.,  AND P. S.  RAND.  1990.  Influence  of substrate  type  on  vulnerability  of  prey to
         predacious aquatic insects. J. N. Am. BenthoLSoc. 9: 1-8.

 GARDNER  W  S.,  T.  F.  NALEPA,  D.  R.  SLAVENS, AND  G.  A.  LAIRD.  1983  Patterns  and
         rates of nitrogen release by benthic chironomidae  and oUgochaeta. Can.  J. Fish AquaL So. 40:
         259-266.                            ••.'•'"•.'.'

  GAUSS   J   D  P  E  WOODS,  R.  W.  WINNER,  AND J.  H. SHILLINGS.  1985.  Acute  toxicity
         of 'copper to .three life stages of efajronomus tengns as affected by water hardness-alkalinity.
         Environ. Pollut (A) 37: 149-157.

  GEROULD   S.,  -P.  LANDRUM,  AND   J.  P.  GffiSY.   1983,  Anthracene  bioconcentration  and
         bidtransformation in chironomids: effects of temperature and concentration. Environ. Pollut 30:
          175-188.                ;                        :

  GESY  J.   P., AND  R. A.  HOKE.  1989.  Freshwater  sediment  toxicity   bioassessment:   rationale
          for species selection and test design. J. Great Lakes Res. 15:  539-569.
                                                      509

-------
GffiSY  J.  P.,  S.  M.  BARTELL, P.  F.  LANDRUM,  G.  J.  LEVERSEE,  AND J.  W. BOWLING.
        1983. Fates and biological effects of polycyclic aromatic hydrocarbons in aquatic systems. In EPA
        Project Summary,, EPA-600/S3-83-053 Nov.,1983.
                                     " 111,                   '         "         .

GffiSY  J.  P., R.  L. GRANEY,  J.  L, NEWSTED,  C.  J.  ROSIU, A.  BENDA,  R.  G.  KREIS  JR.,
        AND F. J. HORVATH. 1988. Comparison of three sediment bioassay methods using Detroit River
        sediments. Environ. Toxicol. Chem. 7: 483-498.

GIESY J. P., C. J. ROSIU, R. L. GRANEY, AND M. G. HENRY. 1990. Benthic invertebrate bioassays with toxic
        sediment and pore water. Environ. Toxicol. Chem. 9: 233-248.

GRANELJ  W.  1979. The  influence  of Chironomus plumosus  larvae on  the  exchange of  dissolved
        substances between sediment and water. Hydrobiologia 66: 149-159.  .

HAND  V. C,  R.  A.  RAPAPORT,  AND  C. A.  PHTINGER.  1990.  First validation of  a model
        for the adsorption of linear alkylbenzenesulfonale (LAS) to sediment and comparison to chronic
        effects data. Chemosphere 21: 741-750.

HARE  L.,   E.   SAOUTER,  P.   G.  C.   CAMPBELL,  A.   TESSIER,  F.  RffiEYRE,  AND  A.
        BOUDOU.  1991. Dynamics of cadmium, lead, and zinc exchange between  nymphs of the
        burrowing mayfly Hexagenia rigida (Ephemeroptera) and the environment Can. J. Fish Aquat Sci.
        48:39-47.

HARREL  R.  C.,  AND  M.  A.  HALL  IE.  1991.  Macrobenthic community   structure before  and
        after pollution abatement in the Neches River estuary (Texas). Hydrobiologia 211: 241-252.

HATAKEYAMA  S.  1987.  Chronic  effects  of  Cd  on  reproduction   of   Polypedilum   nubifer
        (Chironomidae) through water and food. Environ. Pollut 48: 249-261.

HATAKEYAMA S.,  AND  M.  YASUNO.  1987.  Chronic effects  of Cd on  the  reproduction  of  the
        guppy (Poecilia reticulata) through Cd-accumulated midge larvae (Chironomus  yoshimatsui).
        Ecotox. Environ. Safety 14:  191-207.

HEINZ  G. H.,  S.  D. HASELTINE, W. L. REICHEL,  AND  G.  L. HENSLER.  1983. Relationships
        of environmental contaminants to reproductive success in Red-Breasted Mergansers Mergus senator
        from Lake Michigan. Environ. Pollut. (A) 32: 211-232.
HERSHEY  A.  E.  1987.  Tubes  and  foraging  behavior  in  larval  Chironomidae:
       predator avoidance. Oecologia 73: 236-241.
implications  for
HESSLEIN R.  H.,  M.  J.  CAPEL, D.  E.  FOX, AND K.  A. HALLARD.  1991.  Stable  isotopes  of
       sulfur, carbon, and nitrogen as  indicators of trophic level and fish migration in the lower
       Mackenzie River basin, Canada. Can. J. Fish Aquat Sci. 48: 2258-2265.

HOFFMAN  D.  L,  B.  A.  RATTNER,   L.  SILEO,  D.  DOCHERTY,  AND  T. J.  KUBIAK.  1987.
       Embryotoxicity, teratogenicity, and aryl hydrocarbon hydroxylase activity in Fors.ter's Terns on
       Green Bay, Lake Michigan. Environ. Res. 42: 176-184.

HOKE R. A., J.  P.  GIESY,  AND  J. R. ADAMS.  1990a. Use  of linear  orthogonal contrasts  in
       analysis of environmental data. Environ. Toxicol. Chem. 9: 815-819.
                                                  510

-------
INGERSOLL   C.   G.,  AND  M.   K.  NELSON.   Testing  sediment  toxicity  with   Hvalella  azteca
       (Amphipoda) and Cfaironomus rioarius  (Diptera).  Aquatic Toxicology and Risk Assessment
       Thirteenth Volume, ASTM STP  1096, W.G. Landis and W.H. van der Schalie, Eds., Amencan
       Society for Testing and Materials, Philadelphia, 1990, pp. 93-109.

INGERSOLL   C.  G.,  M.  K. NELSON,  AND  F.  J. DWYER.  Proposed  guidelines  for  conducting
       sediment bioassays with midge fChironomus. Chironomidae, Diptera). ASTM, E 47.03, Draft No.
        1.                               '            .                   ,

INGERSOLL   C  G.,  F.  J. DWYER, AND T. W. MAY, 1990.  Toxicity  of inorganic and organic
        selenium to Daohnia magna (Cladocera) and Chironomus rioarius (Diptera). Environ, Toxicol.
        Chem. 9:  1171-1181.                          '              ,

EVEKOUA E. I  1971. On the feeding habits of chironomid larvae. JJmnologica 8: 201-202.

JOHNSON  M.   G.,  O.   C. MCNEIL,  AND  S.  E.   GEORGE   1987   Benthic   m^mvertebrate
        associations in relation to environmental factors in Georgian Bay. J. GreatLakes Res. 13: 310-327.

JOHNSON  R.   K.,  B.   BOSTROM,  AND  W.   VAN  DE   BUND.  1989.  Interactions  between
        Chironomus olumosus (L.) and the microbial community in surficial sediments of a shallow,
        eutropmc lake. LimnoL Oceanogr. 34: 992-1003.

 KAJAK  Z.,  AND  J.   WARDA.   1968.  Feeding  of benthic  non-predatory  Chironomidae  in  lakes.
       • Annales Zoologici Fennici 5:  57-64.

 KAWATSKI  J   A   AND  A.  E.  ZTTTEL.  Accumulation,  elimination  and  biotransformation  of  the
        Uimpricidc r. 5 rtf-M-r^-™*™"^1-"™1^- hv Chironomus tentans. Bureau of Sport Fisheries
        and Wildlife, Investigations in Fish i Control No  79, United States Fish and Wildlife Service, Fish
        Control Laboratory, LaCrosse, WI. ;8 p.

 KHANGAROT  B. S.,  AND P. K.  RAY.  1989.  Sensitivity  Of  midge  larvae of  Chironomus teams
        Fabricius CDiptemChironbmidae) to heavy metals. BuU. Environ. Contain. Toxicol. 42: 325-33U.

 KNEZOVICH J.  P.,   AND'  F.   L.! HARRISON.  1988.  The bioavailabffity  of  setoent-sorbed
                      tn io^^ nf th^. tni.ipft. Chironomus decorus. Ecotox. Environ. Safety 15; 226-241.
  KNEZOVICH  J.  P.,  F.  L.  HARRISON,  AND  R.  G. WJLHELM.   1987.  The  bioavailabmty  of
         sediment-sorbed organic chemicals: a review. Water, Air, Soil PolluL 32: 233-245.

  KOKKINN  M. J.  199k Is the rate of embryonic  development  a  predictor  of  overall development
         rate in Tanvtarsus barbitarsis (Diptera: Chironomidae)? Aust J. Mar. Freshwater Res. 44; 575-579.

  KOSALWAT  P.,  AND  A.  W.  KNIGHT.   1987a.  Acute  toxicity  of  aqueous  and  substrate-bound
         copper to the midge, Chironomus decorus. Arch. Environ. Contam. Toxicol. 16: 275-282.

  _-.  1987b.  Chronic  toxioity  of copper to  a partial  life cycle  of the midge,  Chironomus decorus.
         Arch. Environ. Contam. Toxicol. 16: 283-290.

  KRANTZBERG  G.   1989.   Accumulation   of  essential and  nonessential   metals  by   chironomid
         larvae in relation to physical and chemical properties of the elements. Can. J. Fish Aquat So. 46:
          1755-1761.

  KRANTZBERG G., AND  P. M. STOKES. 1988a. The importance of surface adsorption and  pH in metal
          accumulation by chironomids. Environ. Toxicol. Chem. 7: 653-670.
                                                       511

-------
—.  1990.  Metal  concentrations and  tissue distribution  in larvae  of Chironomus  with  reference  to
       X-ray microprobe analysis. Arch. Environ. Contain. Toxicol. 19: 84-93.

KRAUS M. L. 1989. Bioaccumulation of heavy metals in pre-fledgling tree, swallows, Tachycineta bicolor. Bull.
       Envkon. Contam. Toxicol. 43: 407-414.

KRIEGER  K.  A.   1984.  Benthic  macroinvertebrates  as  indicators  of  environmental  degradation
       in the southern nearshore zone of the central basin of Lake Erie. J. Great Lakes Res. 10: 197-209.

KRUMGALZ B.  S.,  G. FAINSHTE1N,  M.  SAHLER,  AND  L. GORFUNKEL. 1989. 'Field  Error'
       related to marine sediment contamination studies. Mar. PolluL Bull 20: 64-69.

LANDRUM P. R,  AND  R. POORE.  1988.  Toxicokinetics  of  selected  xenobiotics  in  Hexagenia
       limbata. J.  Great Lakes Res. 14: 427-437. •
LARSON  R.  A.,  AND  M.  R. BERENBAUM.  1988. Environmental  phototoxicity:  solar  ultraviolet
       radiation affects the toxicity of natural and man-made chemicals. Envkon. Sci. Technol. 22: 354-
       360.

LARSSON  P.,  L.  OKLA, S.-O.  RYDING,  AND   B. WEST65.  1990.  Contaminated  sediment  as
       a source of PCB's in a river system. Can. J. Rsh Aquat Sci. 47: 746-754.

LENAT D.  R. 1983. Chironomid taxa richness:   natural variation  and  use  in  pollution  assessment
       FreshwaL Invertebr. Biol. 2: 192-198.

LEVERSEE  G.  J., J. P.  GffiSY,  P.,  F.  LANDRUM,  S.  GEROULD, J.  W.  BOWLING,  T.  E.
       FANNIN, J. D. HADDOCK, AND S. M. BARTELL. 1982. Kinetics and biotransformation of
       benzo(a)pyrene in Chironomus riparius. Arch. Environ. Contam. Toxicol. 11: 25-31.  ,

LEVERSEE  G.  S.,  J.  P.  GIESY,  P.  F.  LANDRUM,  S.  GEROULD, J.  W.  BOWLING,  T.  E.
       FANNIN, J. D. HADDOCK, AND S. M. BARFELL. 1982. Kinetics and biotransformation of
       Benzo(a)pyrene in chkonomus riparius. Arch. Envkon. Contam. Toxicol. 11: 25-31.   >

LOBEL P.  B.,  S.  P.  BELKHODE,  S.  E.  JACKSON,  AND  H. P. LONGERJCH.  1991.  Sediment
       in the intestinal tract  a potentially serious source of error in aquatic biological monitoring
       programs. Mar. Environ. Res. 31:  163-174.

LODEN  M.  S.   1974.  Predation   by  chironomid   (Diptera)  larvae  on   oligochaetes.  Limnol.
       Oceanogr. 19: 156-159.

LOHNER  T.  W., AND  W.   J. COLLINS.  1987.   Determination of uptake rate constants for  six
       organochlorines in midge larvae. Environ. Toxicol. Chem. 6: 137-146.            '

LOHNER  T. W., AND  S. W. FISHER.  1990. Effects of  pH  and  temperature  on the acute toxicity
       and uptake of carbaryl in the midge, Cfakonomus riparius. Aquat ToxicoL 16: 335-354.

LONG  E.  R.,  M.  F.  BUCHMAN,   S.  M.  BAY,  R.  J.  BRETELER,  R.   S.  CARR,  P.  M.
       CHAPMAN, J. E. HOSE, A. L. IISSNER, J. SCOTT, AND D. A. WOLFE. 1990. Comparative
       evaluation of five toxicity tests 'with sediments  from San Francisco Bay and Tomales Bay,
       California Environ. Toxicol. Chem. 9: 1193-1214.

LOZANO  S. J.,  S.  L.  O'HALLORAN,  K.  W.  SARGENT,  AND  J.  C.  BRAZNER.  1992. Effects
       of esfenvalerate on aquatic organisms in littoral enclosures.  Environ. ToxicoL Chem. 11: 35-48.
                                                 512

-------
LUGTHART   G.   J.,  B.  WALLACE,  AND  A.  D.   HURYN.  1990.  Secondary  production   of
       chironomid communities in insecticide-treated and untreated headwater streams. Freshwater Biol.
       24:417-427.                                                            ,     •     •

LYDY M.  J., J. T.  ORJS,  P.  C.  BAUMANN, AND  S.  W. FISHER.  1992.  Effects  of sediment
       organic carbon content on the elimination rates  of neutral lipophilic compounds in the midge
       (Chironomus riparius). Environ. Toxicol. Chem. 11; 347-356.                                   ^
                     '*.    ~  ,i      '•'...'
MACEK   K.  J.   1968.   Reproduction   in  brook   trout   (Salvelinus   fontinalis)  fed   sublethal
       concentrations of DDT. J. Fish. Res. Bd. Can. 25: 1787-1796.
       ....                        , r .            •        •'  .      ..       \    •   ,   .  • -
MACKEY A. P. 1977. Growth and development of larval Chironomidae. Oikos 28: 270-275.

MAIER   K.   L,  P.   KOSALWAT,   AND   A.   W.   KNIGHT.   1990.  Culture  of   Chironomus
        decorus (Diptera:Cfaironomidae) and the effect of temperature on its life history. Environ. Entomol.
        19: 1681-1688.                        .                                                '

MASON  W,  T. Cruronomidae  (Diptera)  as  Biological  Indicators  of Water  Quality.   .Symposium
        on Organisms and Biological Communities as Indicatbrs of Environmnetal Quality.                 •

MATISOFF  G.,  J.  B.  FISHER,  AND S.  MATIS.  1985.  Effects  of bentbic macroinvertebrates  on
        the exchange of solutes between sediments and freshwater. Hydrobiologia 122: 19-33.

MCCAHON   C.  P.,   AND  D.   PASCOE.  1988.   Culture   techniques   for   three  freshwater
        macroinvertebrate species and their use in toxicity tests. Chemosphere 17: 2471-2480.

 —   1991.  Brief-exposure  of first  and  fourth   instar  Chironomus  riparius  larvae  to  equivalent
        assumed  doses of cadmium:  effects on adult emergence. Water, Air, Soil PolluL 60: 395-403.

 MCCAIN  B.  B.,  D.  W.  BROWN, M.  M.  KRAHN,  M.   S.  MYERS,  R.  C. CLARK  JR., S. L.
        CHAN, AND D. C. MATINS.'1988. Marine pollution problems, North American West Coast
        Aquat-Toxicol. 11: 143-162.

 MCCAULEY  V. J. E.  1974. Instar  differentiation in  larval chironomidae (Diptera). Can.  Entomol.
      .   106: 179-200.                               .           ,

 MCELRAVY  E. P.,  G.  A.  LAMBERTI,  AND  V.  H.  RESH.  1989. Year-to-year  variation  in  the
         aquatic macromvertebrate fauna of a northern California stream. J. N. Am. Benthol. Soc. 8: 51-63.

 MCELROY  A.   E   1990.   Polycyclic  aromatic hydrocarbon  metabolism  in   the polychaete Nereis
         virens. Aquat ToxicoL 18:  35-50.

 MCLACHLAN   A.   J.   1988.  Refugia   and   habitat   partitioning   among   midges  (Diptera:
         Chironomidae) in rain-pools. Ecol. Entomol. 13: 185-193.      ,       •.

 MELZIAN  B.  D.,  AND  N.  JAWORSKI. 1991. Toxicity of  chlorine  and ammonia  to  aquatic life:
         chemistry, water quality criteria, recent research, and recommended future research. Water Quality
         Standards for the 21st Century: 127^138.

  MOORE  J.  W.  1980.  Factors  influencing   the  composition,' structure and density  of  a population
         of benthic invertebrates. Arch. HydrobioL 88: 202-218.
                                                   513

-------
MUIR  D. C.  G.,  N. P. GRIFT, B.  E.  TOWNSEND, D.  A. METNER, AND W. L. LOCKHART.
       1982. Comparison of the uptake and bioconcentration of fluridone and terbutryn by rainbow trout
       and Chironomus tentans in sediment and water systems. Arch. Environ. Contain. Toxicol. 11: 595-
       602.

MUIR  D.  C.  G.,  B.  E.   TOWNSEND,   AND  W.  L.  'LOCKHART.  1983.  Bioavailability  of  six
       organic chemicals to Chironomus tentans larvae in sediment and water. Environ. Toxicol. Chem.
       2:269-281.                    ^

NEBEKER   A.   V.,   M.   A.  CAIRNS,   J.   H.   GAKSTATTER,  K.   W.   MALUEG,   G.  S.
       SCHUYTEMA, AND D. F. KRAWCZYK. 1984a. Biological methods for determining toxicity of
       contaminated freshwater sediments to invertebrates. Environ. Toxicol. Chem. 3: 617-630.

NEBEKER  A.  V., M.  A.  CAIRNS,  AND- C.  M.  WISE.  1984b.  Relative  sensitivity  of  Chironomus
       tentans life stages to copper. Environ. ToxicbL Chem. 3: 151-158. •

NEBEKER   A.  V.,   S.  T.  ONJUKKA,   AND   M.   A.   CAIRNS.   1988a.   Chronic   effects  of
       contaminated sediment on  Daphnia magna and Cfaironomus tentans. BulL Environ. Contain.
       Toxicol. 41: 574-581.

NEFF  J. M.,  B. W.  CORNABY,  R. M.  VAGA,  T. C.  GULBRANSEN, J. A.  SCANLON,  AND
       D. J. BEAN. 1988a. An Evaluation of the Screening Level Concentration Approach for Validation
       of Sediment Quality Criteria for Freshwater and Saltwater Ecosystems, p. 115-127. In W. J.
       Adams, G. A. Chapman, and W. E. Landis [edsj, Aquatic Toxicology and Hazard Assessment,
       ASTM STP 971, American Society of Testing and Materials.

—.  1988b.  An  evaluation  of the  screening  level  concentration  approach  for  validation of  sediment
       quality criteria for freshwater and saltwater ecosystems. Aquat Toxicol. 10: 115-127.

NELSON  M.   K.,  C.   G.   INGERSOLL,  AND F.   J.  DWYER.  Standard  guide  for  conducting
       sediment toxicity tests with freshwater invertebrates, ASTM subcommittee E-47.03, Draft #6.

NOLTE  U.   1990.  Chironomid  biomass   determination  from  larval  shape.  Freshwater  BioL   24:
       443-451.                                                                             :

NOVAK   M.   A.,  A.   A.   REILLY,  AND  S. J.  TACKLING.  1988.   Long-term   monitoring  of
       polychlorinated biphenyls in  the Hudson River  (New York) using caddisfly larvae and other
       macroinvertebrates. Arch. Environ. Contain. Toxicol. 17: 699-710.

•NOVAK  M.  A., A.  A.  REILLY,  B. BUSH/AND  L. SHANE.  1990.  In  Situ  determination of PCB
       congener-specific first order absorption/desorption rate constants using Chironomus tentans larvae
       (Insecta: diptera: chironomidae). Water Res. 24: 321-327.

OLIVER D. R. 1971. Life history of the Chironomidae. Ann. Rev. EntomoL 16: 211-225.

PALAWSKI D. U.,  J.  B.  HUNN, D. N. CHESTER, AND R.  H. WIEDMEYER  1900.  Interactive
       effects of acidity and aluminum exposure on the life  cycle of the midge Chironomus riparius
       CDiptera). J. 5: 155-162.

PASCOE  D.,  K. A. WILLIAMS,   AND  D.  W.  J.  GREEN.  1989.  Chronic  toxicity  of  cadmium to
       Chironomus riparius Meigen-effects upon larval development and adult emergence. Hydrobiologia
       175: 109-115.
                                                514

-------
PASCOE D   A  F.  BROWN,  B. M. J.  EVANS,  AND  C.  MCKAVANAGH.  1990. Effects  and
       fotr n'f nflmiiTT ^""™ • tป^ซซy *** with r.hironomus rioarius-the influence of food and artificial
       sediment Arch. Environ. Contam. ToxicoL 19: 872-877.

PAYNE  B.   S.,  AND  A. C.  MILLER.  1991.  The  structure  of dominant  invertebrate  assemblages
       in a small southeastern stream. J. 6: 257-266.

PINDER L. C. V. 1986. Biology of freshwater Chironomidae. Ann. Rev. Entomol. 31: 1-23.

prrnNGER  c   A.,  D.  M.   WOLTERING,  AND  J.   A.   MASTERS.  1989.  Bioavaiiabiiity   of
        sediment-sorbed and aqueous surfactants to Chironomus ri^rius (midge). Environ. ToxicoL Chem.
        8: 1023-1033.

POWLESLAND  C.,  AND J. GEORGE.  1986.  Acute .and  Chronic  toxicity  of nickel  to  larvae  of
        Chironomus rioarius (Meigen). Environ. PolluL (A) 42; 47-64.

RAE J. G.  1987.  The  effects, of  flooding and sediments  on  the  structure  of a stream midge
        assemblage. Hydrobiologia 144:  3-10.

ROBACK   S   S    1981   The   immature   chironomids   of  the  eastern  United  States.   V.
        Pentaneurini-Thienemannimvia group. Proceedings of the Academy ,of Natural Sciences  of
        Philadelphia. 133:'73-128.

 ROSIU  C  J  J  P  GIESY, AND  R.  G. KREIS JR.  1989a,  Toxicity  of vertical  sediments in  the
        Trenton Channel, Detroit River, Michigan, to  Chironomus tentans (fosecta:  Chironomidae). J.
        Great Lakes Res. 15: 570-580.

 SAMANT  H S  K. G. DOE, AND, O.  C. VAIDYA. 1990.  An  integrated  chemical and biological
     •    study of the bioavailability  of  metals in sediments from two contaminated harbours m New
         Brunswick, Canada. The Science of the Total Environment 96: 253-268.

 SANDERS   H  O   Sublethal effects  of toxaphene on Daphnids,  Scuds, and  Midges.   Final report
         No. EPA-IAG-141 (D), U.S. EPA, Environmental Research Laboratory, Duluth, MN, p.16.

 SOSTER F. M.,  AND  P.  L.  MCCALL.  1990.  Benthos  response  to  disturbance  in western  Lake
         Erie: field experiments. Can. J. Fish AquaL Sci. 47: 1970-1985.

 STEWARD  A.  R.,  C.  KANDASWAMI,  S.  CHIDAMBARAM,, C.  ZIPER,  J.  P.  RUTKOWSKI,
         AND H. C. SIKKA. 1990. Disposition andmetabolic fate of benzo[a]pyrene m the brown bullhead.
         Environ. Toxicol. Chem. 9: 1503-1512.

  SWINDOLL C.  M.,  AND F.  M.  APPLEHANS.   1987.  Factors  influencing  to  afcumdation  of
         sediment-sorbed hexachlorobiphenyl by midge larvae. Bull. Environ. Contam. ToxicoL 39:1055-
         1062.      .                                               ;

  THURSTON R.  V.,  T.  A. GILFOIL, E.  L.  MEYN, R.  K. ZAJDEL,  T. L  AOKI,  AND G. D.
         VEITH. 1985. Comparative toxicity often organic chemicals to ten common aquatic species. Water
         Res. 19: 1145-1155.                        '

  TIMMERMANS  K.  R., E.  SPDKERMAN,  M.  TONKES,   AND  H.  COVERS.   1992.   Cadmium
         and zinc uptake by two species of aquatic invertebrate predators from dietary and aqueous sources.
         Can. J. Fish Aquat Sci. 49: 655-662.                 .                        •
                                                 515

-------
 VAN  DE GUCHTE  C,  AND  J.  L. MAAS-DIEPEVEEN.  1987.  Screening  sediments  for  toxicity:
        a water-concentration related problem. Proc. Ann. AquaL Tox. Woikshop 14.

 VAN  DE  GUCHTE  C.,  AND  G.  NIEBEEK.  Bioaccumulation  of chlorobenzenes  by  chironomids
        in water and in  sediment-water systems.  Poster presentation  at the 14th Annual Toxicity
        Workshop, November 1-4, 1987, Toronto, Ontario, Canada.

 VAN  DE GUCHTE  C.,  AND  G.  VAN URK.  Discrepancies  in the effects  of  field and  artificially
        heavy metal contaminated aquatic sediments upon midge larvae. Proceedings of Heavy Metals in
        the Environment, Seventh International Conference, September 12-15,1989, Geneva, Switzerland.

 VAN  DE  GUCHTE  C.,  J.  L.  MAAS-DIEPEVEEN,   AND  L.  GROOTELAAR.  Midge  larvae  in
        sediment ecotoxicology. Proceedings of the First European Conference on Ecotoxicology, October
        17-19, 1988, Copenhagen, Denmark.
VAN   DE  GUCHTE  C.,  G.   NIEBEEK,   AND  J.  BOTTERWEG.  Chironomids'   uptake
        chlorobenzenes  from contaminated  sediments.   Proceedings of  the  Second International
        TNO/BMFT Conference on Contaminated Soil, April 11-15, 1988, Hamburg, Germany.
             of
VAN  HATTUM  B.,  K.  R.  TIMMERMANS,  AND  H.  A.  COVERS.  1991.  Abiotic  and  biotic
        factors influencing in  situ  trace metal levels in macroinvertebrates in freshwater ecosystems.
        Environ. Toxicol. Chem. 10: 275-292.

VOGT  N.  B.  1989.  Polynomial   principal  component  regression:    an  approach  to  analysis and
        interpretation of complex mixture relationships in multivariate environmental data. Chemometrics
        and Intelligent Laboratory Systems. 7: 119-130.

WARWICK   W.  F.   1985.  Morphological   abnormalities   in  Chironomidae   (Diptera)   larvae   as
        measures of toxic stress in freshwater ecosystems:  indexing antenna! deformities in Chironomus
        Meigen. Can. J. Fish AquaL ScL 42: 1881-1914.

—.  1990a. Morphological  deformities  in  Chironomidae  (Diptera)  larvae  from  the  Lac  St.  Louis
        and Laprairie Basins of the St. Lawrence River. J. Great Lakes Res. 16: 185-208.

—.  The  use  of  morphological  deformities  in  chironomid  larvae  for biological  effects  monitoring.
        National Hydrology Research Institute,  Saskatoon, Saskatchewan.  NHRI paper no.43.  IWD
        Scientific Series no. 173.

—.   1991.   Indexing   deformities   in  ligulae   and  antennae  of   Procladius     larvae   (Diptera:
        Chironomidae): application to contaminant-stressed environments. Can. J. Fish Aquat ScL 48:
        1151-1166.

WARWICK  W.  F.,  AND   N.   A.   TISDALE.   1988.  Morphological   deformities   in  Chironomus.
        Cryptochironomus. and Procladius larvae (DipteraiChironomidae) from two differentially stressed
        sites in Tobin Lake, Saskatchewan. Can. J. Fish AquaL Sci. 45: 1123-1144.

WEBBER  E.  C.,  D.  R.  BAYNE, .AND W.  C.   SEESOCK.  1989a.  DDT  contamination  of benthic
        macromvertebrates and sediments from tributaries of Wheeler Reservoir, Alabama. Arch. Environ.
        Contam. ToxicoL 18: 728-733.
      1989b.   Macroinvertebrate   communities   in   Wheeler   Reservoir   (Alabama)
        prolonged exposure to DDT contamination. Hydrobiologia 183: 141-155.
tributaries   after
                                                     516

-------
WELCH  H.   E.,  J.  K.   JORGENSON,  AND  M.  F.  CURTIS.  1988.   Measuring  abundance  of
       emerging Chironomidae (Diptera):  Experiments on trap size and design, set duration, and
       transparency. Can. J. Fish Aquat Sci. 45: 738-741.                                              ป

WELTON J.  S., M.  LADLE,  J.  A.  B. BASS, AND  R.  T. CLARKE.  1991.  Grazing of epffithic
       chirpnomid larvae at two different water velocities in recirculating streams. Arch. Hydrobiol. 121:
       405-418.                                                   '                  ,

WHITMAN  R.  L.  1989.   A new sampler for  collection of  interstitial  water  from sandy  sediments.
       Hydrobiologia 176/177: 531-533.

WILEY  M.  J.  1981.   Interacting  influences  of  density and  preference on the  emigration  rates  of
        some lotic chironomid larvae (DipteraChironomidae). Ecology 62: 426-438.

WILLIAMS   K.  A.  1987.  Effect of   cadmium  on  opposition  and  egg  viability  in Chironomus
        rioarius (DipteraiChironomidae). Bull. Environ. Contam. ToxicoL 38: 86-90. (needs to have
        additional authors added to reference)     .

WILLIAMS   K.  A.,  D.  W.  J.  GREEN,  D.  PASCOE,  AND  D.  E.  GOWER.  ,1986.  The acute
        torticity "f ซ"<"'ซ"" *" HifRBrsnt larval stages of Cfaironomus riparius (DipteraChironomidae) and
        its ecological significance for pollution regulation. Oecologia 70: 362-366.

WINNELL  M.  H.,   AND D.  J.   JUDE.   1984.  Associations  among   Chironomidae  and  sandy
        substrates in nearshore Lake Michigan. Can. J. Fish Aquat.  Sci. 41: 174-179.

 WINNELL  M.  H.,  AND  D.  S.   WHITE.   ????  Ecology  of  some  Chironomidae (Diptera)  from
        southeastern Lake Michigan, U.S.A. Trans. Amer. Entomol. Soc. Ill: 279-359.

 WOOD  L.  W.,  G.' Y. RHEE, B.  BUSH, AND E.  BARNARD.  1987. Sediment desorption of PCB
        congeners and their bio-uptake by dipteran larvae. Water Res. 21: 875-884.

 WOODS  P.  E., J. D. PAULAUSKIS,  L. A. WEIGHT, M.  A. ROMANO,  AND  S.  I. GUTTMAN.
         1989. Genetic variation in laboratory and field populations of me midge, Chironomus tentans Fab.:
        implications for toxicology. Environ. Toxicol. Chem. 8: 1067-1074.

 YOUNG L  B.,  AND H.  H.  HARVEY.  1991. Metal concentrations  in  chironomids  in  relation  to
         the geochemical characteristics  of surficial sediments. Arch. Environ. Contam. Toxicol. 21: 202-
         211.                                                     ,

 YOUNT J.  L.  1966.  A  method for  rearing  large  numbers of  pond midge larvae,  with estimates
         of productivity and standing crop. Amer. Midi. Nat -76: 230-238.

 ZEGENFUSS  P.  S.,  W.   J.  RENAUDETTE,  AND  W. J.  ADAMS.  1986.  Methodology  for
         Assessing  the Acute Toxicity of Chemicals Sorbed to Sediments:  Testing the Equilibrium
         Partitioning Theory, p. 479-493. In Aquatic Toxicology and Hazard Assessment, ASTM STP 921,
         American Society of Testing and Materials.

  ZULLO  S.   J,   AND  J.  B.  STAHL.  ????  Abundance,  distribution and  life  cycles   of  midges
         (ChironomidaeiDiptera) in an acid strip-mine lake in Southern Illinois. Amer. Midi. Nat 119:353-
         365.
                                                 517

-------

-------
Group: C:\REF\AMPPSCH1.GRP
Temporary group for searching
 Sorted by: Authors, Year, Title
 Using Format: LMNOL OCEANOGR
 Current Search: term=ampp
 Last Search run on 9 Sep 1992, at 11:57
 Last modified on 9 Sep 1992, at 12:26
Listed with Format LMNOL OCEANOGR

Listing Created 9 Sep 1992, at 12:27
ABEL T., AND F. BARLOCHER. 1988. Uptake of cadmium by Gammarus fossanmi (Amphipbda) from food and
       ' water. J. AppL Ecol. 25: 223-231.

ADAMS J., P. J. WATT, C. J. NAYLOR, AND P. J. GREENWOOD. 1989. Loading constraints, body size and
       mating preference in Gammarus species. Hydrobiologia 183; 157-164.

AHSANULLAH M., AND T.  FLORENCE. 1984. Toxicity of copper to the marine amphipod Allorchestes
       coinpressa in the presence of water-and lipid-soluble ligands. Mar. Biol. 84: 41-45.

ANDERSON  R. L., AND P. SHUBAT.  1984. Toxicity of Flucythrinate to  Gamrharus lacustris, Pteronarcys
       dorsata, and Bracfavcentrus americanus:  importance of exposure duration. Environ. Pollut (A) 35:
       353-365.   .   .'  '  .                                                           •    '     '

ARTHUR J. W., AND E. N. LEONARD.  1970. Effects of copper on Gammarus pseudolimnaeus, Phvsa integra,
       and Campeloma decisum in soft water. J. Fish. Res. Bd. Can. 27: 1277-1283.

ARTHUR J. W-, C. W. WEST,  K. N. ALLEN, AND S. F. HEDTKE. 1987. Seasonal toxicity of ammonia to five
       fish and nine invertebrate species. Bull. Environ. Contain. ToxicoL 38: 324-331.

BARLOCHER F.,  AND C.  W. PORTER.  1986. Digestive enzymes and feeding  strategies of three stream
       invertebrates.  J.  N, Am. Benthol. Soc. 5:  58-66.

BARTON D. R. 1989.  Some problems  affecting the assessment of Great Lakes water quality using  benthic
        invertebrates.  J.  Great Lakes Res. 15: 611-622.

BECKER D.  S., G. R.  BILYARD, AND T. C. GINN. 1990.  Comparisons between sediment bioassays and
        alterations of benthic macroinvertebrate assemblages at a marine superfund site: Commencement
        Bay,  Washington. Environ. Toxicol. Chem. 9: 669-685.                              ,

 BONSDORFF E.,  T.  BAKKE, AND A. PEDERSEN. 1990. Colonization of amphipods  and polychaetes to
        sediments experimentally exposed to oil hydrocarbons. Mar. Pollut. Bull. 21: 355-358.

 BORGMANN U.,  AND M. MUNAWAR,  1989. A new  standardized sediment bioassay protocol using the
        amphipod Hyalella azteca (Saussure). Hydrobiologia 188/189; 425-431.

 BORGMANN U., K.  M. RALPH, AND W. P. NORWOOD. 1989. Toxicity test procedures for Hvalella azteca,
        and chronic toxicity of cadmium and pentachlorophenol to H. azteca, Gammarus faciatus, and  ,
        Dapnnia magna. Arch. Environ. Contam. ToxicoL 18: 756-764.                     .
                                                  519

-------
BORGMANN U., W. P. NORWOOD,  AND K. M. RALPH.  1990. Chronic toxicity and bioaccumulation of
       2^,2',5'- and 3,4,3',4'- Tetrachlorobiphenyl and Aroclor 1242 in the amphipod Hyalella azteca.
       Arch. Envkon. Contam. Toxicol. 19: 558-564.

BRANNON J. M., C. B. PRICE, F. J. REBLLY, J. C. PENNINGTON, AND V. A. MCFARLAND. Effects of
       sediment organic matter composition on bioaccumulation of sediment organic contaminants; Interim
       results.  Miscellaneous paper D-91-4, US-Army Engineer Waterways Experiment Station,
       Vicksburg, MS.

BURTON  G. A. Jr., B.  L. STEMMER, K.  L.  WINKS, P. E. ROSS,  AND  L.  C. BURNETT. 1989. A
       multitrophic  level evaluation of sediment toxicity  in Waukegan and Indiana harbors. Environ.
       Toxicol. Chem. 8: 1057-1066.                                                            .

BURTON JR. G. A. 1991. Assessing the toxicity of freshwater sediments. Environ. Toxicol. Chem. 10:1585-1627.

CAIRNS M. A., A. V. NEBEKER, J. H. GAKSTATTER, AND W. L. GRIFFIS. 1984. Toxicity of copper-spiked
       sediments to  freshwater invertebrates. Envkon. Toxicol. Chem. 3: 435-445.        ;

CARIGNAN R., AND D. R. S. LEAN. 1991. Regeneration of dissolved substances in  a seasonally anoxic lake:
       The relative importance of processes occurring in the water column and in the sediments. Limnol.
       Oceanogr. 36: 683-707.

CHAPMAN P. M. 1986. Sediment quality criteria from the sediment quality triad:  an example. Envkon. Toxicol.
       Chem. S: 957-964.

CHAPMAN P. M., R. N. DEXTER, AND E. R. LONG.  1987. Synoptic measures of sediment contamination,
       toxicity and infaunal community composition (the sediment quality triad)  in San Francisco Bay.
       Mar. Ecol. Prog. Ser. 37: 75-96.

CHAPMAN P. M., E. A. POWER, R. N. DEXTER,  AND H. B. ANDERSEN. 1991. Evaluation of effects
       associated with an oil platform, using the sediment quality triad. Envkon. Toxicol. Chem. 10:407-
       424.                                                                                -

CLARK K. E., F. A.  P. C. GOBAS, AND D. MACKAY. 1990. Model of organic chemical uptake and clearance
       by fish from food and water. Envkon. Sci. Technol. 24: 1203-1213.

COOPER W. 1965. Dynamics and production of a natural population of a fresh-water amphipod, Hvalella azteca.
       Ecol Monogr. 35: 377-394.

CROSSLAND N. O., G. C. MITCHELL, AND P. B. DORN. 1992. Use of outdoor artificial streams to determine
       threshold toxicity concentrations for a petrochemical effluent Envkon. Toxicol. Chem. 11:49-60.

DANIELS S. A., M. MUNAWAR, AND C. I. MAYEEELD. 1989. An improved emulation technique for the
       bioassessment of sediment contaminants. Hydrobiologia 188/189: 619-631.

DE MARCH B. G. E, 1977. The effects  of photoperiod and temperature on the induction and termination of
       reproductive resting stage in the freshwater amphipod Hyallela azteca (Saussure). Can. J. Zool. 55:
       1595-1600.

—. 1978. The effects  of constant and variable temperatures on the size, growth, and reproduction of me freshwater
       amphipod Hvallela azteca (Saussure). Can. J. Zool. 56:  1801-1806.
                                                   520

-------
~.  1988. Acute toxicity of binary mixtures of five cations (Cu2*, Cd2*,Zn2+, Mg2*, and K* to the freshwater
       amphipod Gammarus lacustris (Sars): alternative descriptive models. Can. J. Fish Aquat ScL 45:
       625-633.                             .

DEWITT T. H., G. R. DITSWORTH,  AND R. C. SWARTZ.  1988. Effects of natural sediment features on
               of the, phmtncephalid amnhipod. Rheooxynius abronius. Mar. Environ. Res. 25: 99-124
DEWITT T. H, R. C. SWARTZ, AND J. O. LAMBERSON.  1989. Measuring the acute toxicity of estuarine
        sediments. Environ. Toxicol. Chem. 8: 1035-1048.

PEWTTT T.  H.,  R.  J. OZRETICH,  R.  C, SWARTZ, J.  O. LAMBERSON, D.  W.  SCHULTS, G.  R.
        DITSWORTH, J. K. P. JONES, L. HOSELTON, AND L. M. SMITH. 1992. The influence of
        organic matter quality on the toxicity and partitioning of sediment-associated fluoranthene. Environ.
        Toxicol. Chem. 11: 197-208.                                                   ,

DI TORO D. M., J. D. MAHONY, D. J.  HANSEN, K. J. SCOTT, M. B. HICKS, S. M. MAYR, AND M. S.
        REDMOND.  1990. Toxicity of cadmium in sediments:  the role of acid volatile sulfide. Environ.
        Toxicol. Chem. 9: 1487-1502.

DORN P. B., AND J. H.  RODGERS JR.  1989. Variability associated with identification of toxics in national
        pollutant discharge elimination systems (NPDES) effluent toxicity tests. Environ. Toxicol. Chem.
        8: 893-902.                                                          ,   ,

DORN  P. B., R. VAN COMPERNOLLE, C. L: MEYER, AND N. O.  CROSSLAND. 1991. Aquatic hazard
        assessment of the toxic fraction from the effluent of a petrochemical plant Environ. Toxicol.
        Chem. 10: 691-703.

EADIE B. J., P. F. LANDRUM, AND W. FAUST. 1982a. Polycyclic aromatic hydrocarbons in sediments, pore
      ,  water, and the amphipod Pontoporeia hoyi from Lake Michigan. Cbemosphere 11: 847-858.

EATON J.  G., V. R. MATTSON, L. H. MUELLER, AND D. K. TANNER. 1983. Effects of suspended clay on
        bioconcenttation of Kelthane in fathead minnows. Arch. Environ. Contain, Toxicol. 12: 439-445.

EVANS M. S., G. E. NOGUCffl, AND C. P..RICE.ป1991. The biomagnification of polychlorinated biphenyls,
        toxaphene, and DDT compounds in a Lake Michigan offshore food web. Arch. Environ. Contain.
        Toxicol. 20:  87-93.

 FERRARO S. P., H. LEE H, R. J. OZRETICH, AND D. T. SPECHT. 1990. Predicting bioaccumulation potential:
        a test of a fugatity-based model Arch. Environ. Contain. Toxicol. 19: 386-394.

 FRANCE R. L.   1992. Biogeographical variation in size-specific fecundity of the amphipod  Hvalella azteca.
        Crustaceana  62(3):240-248  -

 FRANCE R. L.,  AND B. D. LAZERTE. 1987. Empirical hypothesis to explain the restricted distribution of
        Hvalella azteca in anthropogenically acidified lakes. Can. J. Fish Aquat Sci. 44;. 1112-1121.

 FRANCE R. L., AND P..M. STOKES. 1987a. Influence, of manganese, calcium, and aluminum on hydrogen ion
         tnxicitv to the amphipod Hvalella azteca. Can. J, Zool. 
-------
GANNON J. E., AND A M. BEETON. 1971. Procedures for determining the effects of dredged sediments on
       biota-benthos viability and sediment selectivity tests. JWPCF 43: 392-398.

GARDNER W. S., P. F. LANDRUM, AND J. F. CAVALETTO. 1990. Lipid-partitioning and disposition of
       benzo[a]pyrene and hexachlorobiphenyl in Lake Michigan Pontoporeia hoyi and Mysis relicta.
       Environ. Toxicol. Chem. 9: 1269-1278.

HARGRAVE B. T. 1970a. Distribution, growth, and seasonal abundance ofHyalella azteca (Amphipoda) in relation
       to sediment microflora. J. Fish. Res. Bd. Can. 27: 685-699.

—. 1970b. The utilisation of benthic microflora by Hyalella azteca.        '

HARGRAVE B. T., G. C. HARDING, W. P. VASS, P. E. ERICKSON, B. R. FOWLER, AND V. SCOTT.
       1992. Organochlorine pesticides and polychlorinated biphenyls in the Arctic Ocean food web. Arch.
       Environ. Contain. Toxicol. 22:41-54.                         •           ,            ,

HELL C., AND R. ELMGREN.  1987. Vertical distribution in the sediment in the co-occurring benthic amphipods
      ' Pontoporeia affmis and P. femorata. Oikos 49: 221-229.

INGERSOLL C. G., AND M.  K. NELSON. Testing  sediment toxicity with Hvalella azteca  (amphipoda) and
       Chironomus riparius  (Diptera).  Aquatic Toxicology and Risk Assessment   Thirteenth  Volume,
       ASTM STP 1096, W.G. Landis and W.H. van der Schalie, Eds., American Society for Testing and
       Materials,  Philadelphia, 1990, pp. 93-109.

INGERSOLL C. G., F. J. DWYER, S. A. BURCH,  M. K. NELSON, D..R. BUCKLER, AND J. B. HUNN.
       1992. The use of freshwater and saltwater animals to distinguish between  the toxic effects of
       salinity and contaminants in irrigation drain water. Environ. Toxicol. Chem.  ll: 503-511.

KEMP P. F., AND R. C. SWARTZ. 1988. Acute Toxicity of interstitial and particle-bound.cadmium to a marine
       infaunal amphipod. Mar. Environ. Res. 26: 135-153.

KNEZOVICH J. P., F. L. HARRISON, AND R.  G. WILHELM. 1987. The bioavailability of sediment-sorbed
       organic chemicals: a review. Water, Air, Soil PolluL 32: 233-245.

KRAFT K, J. 1979. Pontoporeia distribution along the Keweenaw shore of Lake Superior affected by copper
       tailings. J. Great Lakes Res. 5: 28-35.

KUDJO DZANTOR E., AND A. S. FELSOT. 1991. Microbial responses to large concentrations of herbicides in
       soil Environ. Toxicol. Chem. 10: 649-655.

LAMBERSON J. O., AND R. C. SWARTZ. Proposed new standard guide for conducting solid phase 10-day static
       sediment toxicity tests  with marine and estuarine amphipods.' USEPA Marine Science Center.
       ASTM Draft. DraftNo. 5.

—. "Standard guide for conducting 10-day  static sediment toxicity tests with marine and estuarine amphipods."
       American  Society for Testing and Materials Designation E 1367-90.

LANDRUM P. F. 1987. Reduction in bioavailability of organic contaminants to the amphipod Pontoporeia hovi by
       dissolved organic matter of sediment interstitial waters. Environ. Toxicol. Chem. 6: 11-20. (needs
       to have additional authors added to reference)

—. 1989. Bioavailability and toxicokinetics of polycyclic  aromatic hydrocarbons sorbed  to sediments for the
       amphipod  Pontoporeia hoyi. Environ. Sci. Technol. 23: 588-595.,
                                                 522

-------
LANDRUM P. R, AND R. POORE. 1988. Toxicokinetics of selected xenobiotics in Hexagenia limbata. J. Great
       Lakes Res. 14:427-437.             .   ;     :  :

LANDRUM P. R, AND W. R. FAUST. Effea of variation in sediment composition on the uptake rate coefficient
       for selected PCB and PAH congeners by the amphipod Diporeia sp. Aquatic Toxicology and Risk
       Assessment Fourteenth Volume, ASTM STP1124, M.A. Mayes and M.G. Barton, Eds., American
       Society for Testing and Materials, Philadelphia, 1991, pp. 263-279.
LANDRUM P. R,  AND C. R.  STUBBLEFIELD. 1991. Role of respiration in the accumulation of organic
        xenobiotics by the amphipod DJEoraa sp. Environ. ToxicoL Chem. 10: 1019-1028.
LANDRUM P. R, M. D. REINHOLD, S. R. NfflART, AND B. J. EADIE. 1985. Predicting the bioavailability
        of organic xenobiotics to Pontoporeia hovi in the presence of humic and fulvic materials and
        natural dissolved organic matter. Environ. ToxicoL Chem. 4: 459-467.

LANDRUM P  R, B. J. EADIE, AND W. R.  FAUST.  1991. Toxicokinetics and toxicity  of a mixture of
        sediment-associated polycyclic aromatic hydrocarbons to the amphipod Diporeia sp. Environ.
        ToxicoL Chem. 10: 35-46.

LANDRUM P., B. EADIE; AND W. FAUST.  1992. Variation in the bioavailability of polycyclic aromatic
        hydrocarbons to the amphipod Diporeia (spp.) with sediment aging. Environ. ToxicoL Chem. 11:
        1197-1208.

 LIEN G.  J., K. E. BIESINGER, L. E. ANDERSON, E. N. LEONARD, AND M. A. GIBBONS. A  toxicity
        evaluation to lower Fox River water and sediments. EPA/600/3-86/008. March 1986.

 LONG E  R.  M R BUCHMAN, S. M- BAY, R. J. BRETELER, R. S. CARR, P. M. CHAPMAN, J. E. HOSE,
        A L LISSNER, J. SCOTT, AND D. A. WOLFE. 1990. Comparative evaluation of five toxicity
        tests with sediments from San Francisco Bay and Tomales .Bay, California. Environ. ToxicoL
        Chem. 9: 1193-1214.

 LOPEZ G., AND R. ELMGREN. 1989. Feeding depths and organic absorption for the deposit-feeding benthic
        amphipods Pontoporeia affins and Pontoporeia femorata. LimnoL Oceanogr. 34: 982-991.

 LYMAN  W J  A E GLAZER, J. H. ONG, AND S. F. COONS. An overview of sediment quality in the United
        States.' Final Report. Contract No. 68-01-6951, task 20. U.S. Environmental Protection Agency-
        Region V. EPA-905/9-88-002.  ChicagoJL.

 LYNCH T. R., AND H. E. JOHNSON.  1982. Availability of a hexachlorobiphenyl isomer to benthic amphipods
        from experimentally contaminated natural sediments, p. 273-287. In J. G. Pearson, R. B. Foster,
         and W. E. Bishop [eds.], Aquatic Toxicology and Hazard Assessment: Fifth Conference, ASTM
         STP 766, American Society for  Testing and Materials.

 MAC M. J., G. E. NOGUCffi, R. L HESSELBERG,  C. C. EDSALL,  J. A. SHOESMTTH, AND J. D.
         BOWKER. 1990. A bioaccumulation bioassay for rreshwater sediments. Environ. ToxicoL Chem.
         9: 1405-1414.           ,

  MACKIE G. L. 1989. Tolerances of five benthic invertebrates to hydrogen ions and metals (Cd,Pb,Al). Arch.
         Environ. Contam. ToxicoL 18: 215-223.
                     )
  MALTBY L. 1992. The use of the physiological energetics of Gammarus pulex to assess toxicity:  a study using
          artificial streams. Environ. ToxicoL Chem. 11: 79-86.
                                                523

-------
 MARCUS W. A. 1989. Regulating contaminated sediments in aquatic environments:  A hydrologic perspective.
        Environ. Management 13: 703-713.

 MAURER D., T. M. CHURCH, C. LORD, AND C. WETHE. 1985. Marine benthos in relation to pore water
        chemistry and sediment geochemistry of simulated dredged material. Int Revue ges. Hydrobiol.
        70:369-377.

 MCCAHON C.  P., AND D. PASCOE. 1988a. Cadmium toxicity to the freshwater amphipod Gammarus pulex
        during the moult cycle. Freshwater Biol. 19: 197-203.

 —. 1988b. Culture techniques  for three freshwater macroinvertebrate species and  their use in toxicity tests.
        Chemosphere 17: 2471-2480.

 —. 1988c. Increased sensitivity to cadmium  of the freshwater amphipod Gammarus pulex (L.) during the
        reproductive period. AquaL Toxicol. 13: 183-194.

 —. 1988d. Use of Gammarus pulex in safety evaluation tests: culture and selection of a sensitive life stage. Ecotox.
        Environ. Safety 15: 245-252.

 MCCARTHY J. R, B. D. JIMENEZ, G. R. SOUTHWORTH, D. M. DITORO, AND M. C. "BLACK. 1986.
        Anomalous binding of organic contaminants may be artifactual due to radiochemical impurities.
        Water Res. 20: 1251-1254.

 MCMURTRY M. J. 1984. Avoidance of sublethal doses of copper and zinc by tubificid oligochaetes. J. Great
        Lakes Res. 10: 267-272.

 MTLSTEAD B.,  AND S. T. THRELKELD.  1986. An experimental analysis of darter predation on Hyalella azteca
        using semipermeable enclosures. J. N. Am. Benthol. Soc. 5: 311-318.

 MUNAWAR M., W. P. NORWOOD, L. H. MCCARTHY, AND C. I. MAYFIELD.  1989. In situ bibassessment
        of dredging and disposal activities in a contaminated ecosystem: Toronto Harbour. Hydrobiologia
        188/189:601-618.

 MUNKITTRICK K. R. 1991.  Calcium-associated reproductive problems  of fish in acidified  environments:
        evolution from hypothesis to scientific fact Environ. ToxicoL Chem. 10: 977-979.

 MUSKO I. B., W. MEINEL, R. KRAUSE,  AND M. BARLAS. 1990. The impact of Cd and different pH on the
        amphipod Gammarus fossarum Koch (Crustacea:  Amphipoda). Comp. Biochem. Physiol. 96a 11- -
        16.

'NEBEKER A. V., AND C. E. MILLER. 1988. Use of the amphipod crustacean Hvalella azteca in freshwater and
        estuarine sediment toxicity tests. Environ. ToxicoL Chem. 7: 1027-1033.

 NEBEKER A. V., M. A. CAIRNS, J. H. GAKSTATTER, K.. W. MALUEG, G, S. SCHUYTEMA, AND D. F.
        KRAWCZYK. 1984. Biological methods for determining toxicity of contaminated freshwater
        sediments to invertebrates. Environ. Toxicol. Chem. 3: 617-630.

 NEBEKER A. V., W. L. GRIFFIS, C. M. WISE,  E.  HOPKINS,  AND L A.  BARBITTA.  1989a. Survival,
        reproduction and bioconcentration in invertebrates and fish exposed to hexachlprobenzene. Environ.
        Toxicol. Chem. 8: 601-611.

 NEBEKER A. V., G. S. SCHUYTEMA, W. L. GRIFFIS, J. A. BARBITTA, AND L. A. CAREY. 1989b. Effect
        of sediment organic carbon on survival of Hvalella azteca exposed to DDT and endrin. Environ.
        Toxicol. Chem. 8: 705-718.
                                                524

-------
NEBEKER A. V., S. T. ONJUKKA, D. G. STEVENS, G. A. CHAPMAN, AND S, E. DOM1NGUEZ. 1992.
       Effects of low dissolved oxygen on survival, growth and reproduction of Dapbniง> Hyalella and
       Gammarus. Environ. Toxicol. Chem. 11: 373-380.

NEFF J. M., B. W.  CORNABY, R. M. VAGA, T. C. GULBRANSEN, J. A.'SCANLON, AND D. J. BEAN.
       1988. An evaluation  of the screening level concentration approach for validation of sediment
       quality criteria for freshwater and saltwater ecosystems: Aquat Toxicol. 10: 115-127.

NELSON M. K., G. G. INGERSOLL, AND F. J. DWYER. Proposed guidelines for conducting sediment bioassays
       with Hvalella azteca (Saussure), Amphipoda.  ASTM Draft No. 1.

—. Standard guide for conducting sediment toxicity tests with freshwater invertebrates, ASTM subcommittee E-
       47.03, Draft #6.

NEWMAN R. M. 1987. Comparison of encounter model predictions with observed size-selectivity by stream trout
       L N. Am. BenthoL Soc. 6: 56-64.

NIPPER M. G., D. J. GREENSTEIN, AND  S. M. BAY. 1989a. Short- and long-term sediment toxicity test
       methods with the amphipod Grandidierella iaponica. Environ. Toxicol. Chem. 8:  1191-1200.

PANTANI C., P. F. GHETTI, A. CAVACINI, AND P. MUCCICONI. 1990. Acute toxicity.of equitoxic binary
        mixtures of some metals, surfactants and pesticides to the freshwater amphipod Gammarus italicus
        Environ. TechnoL 11: 1143-1146.

 PETERSON C. H. 1991. Intertidal zonation of marine invertebrates in sand and mud. American Scientist.79: 236-
        249.             •

 PLESHA P. D., J.  E. STEIN, M. H.  SCHJEWE, B. B. MCCAIN, AND U. VARANASL 1988a.  Toxicity of
        marine sediments supplemented with mixtures of selected chlorinated and aromatic hydrocarbons
        to the infaunal amphipod Rhepoxvnius abronius. Mar. Environ. Res. 25: 85-97.

 QUIGLEY M. A.,  AND G. A. LANG. 1989. Measurement of amphipod'body length using  a digitizer.
        Hydrobiologia 171: 255-258.                                                          ,  ,

 QUIGLEY M A, AND H. A. VANDERPLOEG. 1991. Ingestion of live filamentous diatoms by the  Great Lakes
        amphipod, Diporeiasp.: a case study of me limited value of gut contents analysis. Hydrobiologia
        223:141-148.

 QUIGLEY M. A., J. F. CAVALETTO, AND W. S. GARDNER. 1989. Lipid composition related to size and
        'maturity of the amphipod Pontoporeia hoyi. J. Great Lakes Res. IS: 601-610.

 REICHERT W. L., B. L. EBERHART, AND U. VARANASL 1985. Exposure of two species of deposit-feeding
         amphipods to sediment-associated [3H]Benzo-[a]pyrene: Uptake, metabolism and covalent binding
         to tissue macromolecules. Aquat Toxicol. 6: 45-56.      '

 ROBERTS JR. M. H., W. J. HARGIS JR., C. J. STROBEL, AND P. F. DE LISLE. 1989. Acute toxicity of PAH
         contaminated sediments, to the estuarine fish Leiostomus xanthurus. BuU. Environ.  Contam.
         Toxicol. 42: 142-149.                                                         .
           '                 v   ,       V   '     •
 ROBINSON A- M. 1988. Effects of culture conditions on the sensitivity of a phoxocephalid amphipod, Rhepoxynius
         abronius, to cadmium in sediment Environ. Toxicol. Chem. 7: 953-959.
                                                 .525

-------
ROBINSON A. M., J. O. LAMBERSON, F. A. COLE, AND R. C. SWARTZ. 1988. Effects of culture conditions
       on the sensitivity of a phoxocephalid amphipod, Rhepoxynius abronius. to cadmium in sediment
       Environ. Toxicol. Chem. 7: 953-959.

SANDERS H. O. Sublethal effects of toxaphene on Dapfanids, Scuds, and Midges. Final report no. EPA-IAG-141
       (D), U.S. EPA, Environmental Research Laboratory, Duluth, MN. p.16.

SCHLEKAT C. E., B. L. MCGEE, AND E. REINHARZ. 1992. Testing sediment toxicity in Chesapeake Bay with
       the amphipod Leptocheirus plumulosus: an evaluation. Environ. Toxicol. Chem. 11: 225-236.

SCHUYTEMA G. S.,  D. F. KRAWCZYK, W. L. GRIFHS,  A. V. NEBEKER, M. L. ROBIDEAUX, B. J.
       BROWNAWELL, AND J. C. WESTALL. 1988, Comparative uptake of hexachlorobenzene by
       fathead minnows, amphipods and oligochaete worms from water and sediment Environ. Toxicol.
       Chem. 7: 1035-1045.

SCHUYTEMA G. S., A. V. NEBEKER,  W. L. GREFFIS, AND C. E.  MILLER.  1989. Effects of freezing on
       toxicity of sediments contaminated with DDT and endrin. Environ. ToxicoL Chem. 8: 883-891.

SCHUYTEMA G. S., D. F. KRAWCZYK, W. L. GRIFHS, A. V. NEBEKER, AND M. L. ROBIDEAUX. 1990.
       Hexachlorobenzene  uptake by  fathead minnows  and macroinvertebrates  in recirculaling
       sediment/water systems. Arch. Environ. Contain. Toxicol. 19: 1-9.

SERVOS M. R., AND D. C. G. MUIR. 1989. Effect of dissolved organic matter from Canadian shield lakes on
       thebioavailability of 1,3,6,8-tetrachlorodibenzo-p-dioxin to the amphipod Crangonyx laurentianus.
       Environ. Toxicol. Chem. 8: 141-150.                                        •'•''.

SHAW G. R., AND D. W. CONNELL. 1984. Physicochemical properties controlling polychlorinated biphenyl
       (PCS) concentrations in aquatic organisms. Environ; ScL Technol. 18: 18-23.

SPEHAR R. L., H. P. NELSON, M. J. SWANSON, AND J. W. RENOOS. 1985.  Pentachlorophenol toxicity to
       amphipods and fathead minnows at different test pH values. Environ. Toxicol. Chem. 4:  389-397.

STEPHENSON M.,  AND G. L. MACKIE. 1989. A laboratory study of the  effects of waterborne cadmium,
       calcium,  and  carbonate  concentrations on  cadmium concentrations  in  Hyalella  azteca
       (CrustaceazAmphipoda). AquaL Toxicol. 15: 53-62.

STRONG D. R. Jr. 1972. Life history variation among populations of an amphipod (Hyalella azteca). Ecology 53:
       1103-1111.

SWARTZ R. C.,'G. R. DITSWORTH, D. W. SCHULTS, AND J.  O. LAMBERSON. 1985.  Sediment  toxicity
       to a marine infaunal amphipod: c^minm and its interaction with sewage sludge. Mar. Environ.
       Res. 18: 133-153.

SWARTZ R.  C., P. F. KEMP, D. W. SCHULTS, AND J. O. LAMBERSON.  1988.  Effects of mixtures of
       sediment contaminants on the marine infaunal amphipod, Rhepoxynius abronius. Environ. Toxicol.
       Chem. 7: 1013-1020.

SWARTZ R. C, P. F.  KEMP, D. W.  SCHULTS, G. R DITSWORTH, AND R. J. OZRETICH. 1989. Acute
       toxicity of sediment from Eagle  Harbor, Washington, to the infaunal amphipod Rhepoxvnius
       abronius. Environ. ToxicoL Chem. 8: 215-222.

SWARTZ R.  C., D. W. SCHULTS, T. H. DEWTTT, G. R. DITSWORTH, AND J. O; LAMBERSON. 1990.
       Toxicity of fluoranthene in sediment to marine amphipods: a test of the equilibrium partitioning
       approach to sediment quality criteria. Environ. ToxicoL Chem..9: 1071-1080.
                                              526

-------
SWARTZ R. c, D w. SCHULTS, j. o; LAMBERSON, R. L OZRETICH, AND j. K. STULL.' \m. vertical
       profiles of toxicity, organic carbon, and chemical contaminants in sediment cores from the Palos
       Verdes shelf and Santa Monica Bay, California. Mar. Environ. Res. 31:; 215-225.

THOMANN R  V., J. P. CONNOLLY, AND T. F. PARKERTON. 1992. An equilibrium model of organic
       chemical Accumulation in aquatic food webs with sediment interaction. Environ. ToxicoL Chem.
       11:615-629.

VAN DER KOOY L. A., D. VAN DE MEENT..C J. VAN LEEUWEN, AND W. A. BRUGGEMAN. 1991.
       Deriving quality criteria for water and sediment from the results of aquatic toxicity tests^and
       product standards: application of the equilibrium partitioning method. Water Res. 25: 697-705.

VAN DER OOST R.  H. HEIDA, AND A. OPPERHUEEN.  1988. Polychlorinated biphenyl congeners  in
       sediments plankton, molluscs, crustaceans, and eel in a freshwater lake:  implications of usmg
       reference chemicals and indicator organisms in bioaccumulation studies. Arch. Environ. Contam.
       ToxicoL 17:  721-729.

VARANASIU. 1985. Bioavailability and biotransformation of aromatic hydrocarbons in benthic organisms exposed
        to sediment from an urban estuary. Environ. Sci. Technol. 19: 836-841.

WILLIAMS L  G P M. CHAPMAN, TAND T. C. GINN.  1986a. A comparative evaluation of marine sediment
        toxicity using bacterial luminescence, oyster embryo and amphipod sediment bioassays. Mar.
        Environ. Res. 19: 225-249.

 WINGER P. V., AND P. J. LASER: 1991. A vacuum-operated pore-water extractor for estuarine and freshwater
        sediments. Arch. Environ. Contam. Toxicol. 21: 321-324.
                                                527

-------

-------
Group: C:\REF\OLCHSCH1.GRP                                                 -
Temporary group for searching
  Sorted by: Authors, Year, Tide
  Using Format: LIMNOL OCEANOGR
  Current Search: term=olch or term=olig
  Last Search run on 9 Sep 1992, at 12:07
  Last modified on 9 Sep 1992, at 13:0

Listed with Format LIMNOL OCEANOGR          "        ,

Listing Created 9 Sep  1992, at 13:06                     .


                                                   ••              %   .              '      ' '
ALLER R. C., AND J. Y. YINGST. 1978. Biogeochemistry of tube-dwellings: A study of the sedentary polychaete
        Amphitrite ornata (Leidy). J. Mar. Res. 36; 201-254.

BACK H. 1990. Epidermal uptake of Pb, Cd, and Zn in tubificid worms. Oecologia 85: 226-232.

BAILEY H C  AND D H. W. LIU. "Lumbriculus variegatus, a benthic oligochaete, as a bioassay organism."
         Aquatic Toxicology,  ASTM  STP 707,  J.C. Eaton, PJL Parrish, and A.C. Hendricks, Eds.,
         American Society for Testing  and Materials, 1980, pp. 205-215.

 BARTON D. R.  1989. Some problems affecting  the assessment of Great Lakes water quality using  benthic
         invertebrates. J. Great Lakes Res. 15:  611-622.

 BAUER fflLTY A R. DALLINGER, AND B.  BERGER. 1989. Isolation and partial characterization of a cadmium-
         binding protein from Lumbriculus variegatus (Oligochaeta, Annelida). Comp. Biochem. Physiol.
         94C: 373-379.

 BENSON W H  AND S. F. LONG. 1991. Evaluation of humic-pesticide interactions on the acute toxicity of
         selectedorganophosphate and carbamate insecticides. Ecotox. Environ. Safety 21: 301-307.

  BERGHOUT A G R V., E. WENZEL, J. BULD, AND K. J. NETTER. 1991. Isolation, partial purification, and
         characterization of the cytochrome P-450-dependent monooxygenase system from the midgut of
         the earthworm Lumbricus terrestris. Comp, Biochem. Physiol. lOOc: 389-396.

  BONSDORFFE.,T.BAKKE, AND A.PEDERSEN. 1990. Colonization of amphipods andpolychaetes to sediments
         experimentally exposed to oil hydrocarbons. Mar. Pollut BuU. 21: 355-358.

 - BOON J P  AND J  C DUINKER. 1985. Kinetics of polychlorinated biphenyl (PCB) components in juvenile sole
          (Solea solea) in relation to concentrations in water and to lipid metabolism under conditions of
         .starvation. AquaL Toxicol. 7: 119-134.

  BOUGUENEC V. 1992. Oligochaetes (Tubificidae and Enchytraeidae) as  food in fish rearing:  a review-and
          tffeliminary tests. Aquapulture 102: 201-217.

  BRANNON J M., C. B. PRICE, F. J. REILLY,  J. C. PENNINGTON, AND V. A. MCFARLAND. Effects of
          sediment organic matter composition on bioaccumulation of sediment organiccontammants; Intenm
          results.   Miscellaneous  paper D-9M, US Army Engineer Waterways  Experiment Station,
          Vicksburg,  MS.

  BRINKHURST  R. O. 1989. A phylogenetic analysis of the Lumbriculidae (Annelida, Oligochaeta). Can. J. Zool.
          67: 2731-2739.
                                                   529

-------
 BRINKHURST R. O., AND K. E. CHUA. 1969. Preliminary investigation of the exploitation of some potential
        nutritional resources by three sympatric tubificid oligochaetes. J. Fish. Res. Bd. Can. 26: 2659-
        2668.

 BURTON JR. G. A. 1991. Assessing the toxicity of freshwater sediments. Environ. Toxicol. Chem. 10:1585-1627.

 CARLSON A. R., G. L. PfflPPS, V. R, MATTSON, P. A. KOSIAN, AND A. M. COTTER. 1991. The role of acid-
        volatile sulfide in determining cadmium bioavailability and toxicity in freshwater sediments.
        Environ. Toxicol. Chem. 10:  1309-1319.

 CARR R. S., J. W. WILLIAMS, AND C. T. B. ERAGATA. 1989. Development and evaluation of a novel marine
        sediment pore water toxicity test with the polychaete Dinophilus gvrociliatus. Environ. Toxicol.
        Chem. 8: 533-543.

 CHADWICK G. G., AND R,  W. BROCKSEN. 1969. Accumulation of dieldrin by fish and selected fish-food
        organisms. J.'Wildl. Manage. 33: 693-700.

 CHAPMAN P. M. 1986. Sediment quality criteria from the sediment quality triad:  an example. Environ. Toxicol.
        Chem. 5: 957-964.                                                                       •

 CHAPMAN P. M., AND R, O. BRINKHURST. 1984. Lethal and sublethal tolerances of aquatic oligochaetes with
        reference to their use as a biotic index of pollution. Hydrobiologia 115: 139-144.

 CHAPMAN P. M., AND R. FINK. 1984. Effects of Puget Sound sediments and their elutriates on the life cycle of
        Capitella 'capitata. BulL Environ. Contain. Toxicol. 33: 451-459.

 CHAPMAN P. M., L. M. CHURCHLAND, P. A. THOMSON, AND E. MICHNOWSKY. 1980a. Heavy metal
        studies with oligochaetes, p. 477-502. In R. O. Brinkhurst, and D. G. Cook [eds.], Aquatic
        Oligochaete Biology, Plenum Publishing Corp.

 CHAPMAN P. M., M. A. FARRELL, AND R. O. BRINKHURST. 1982a. Relative tolerance of selected  aquatic
        oligochaetes to combinations of pollutants and environmental factors. Aquat. Toxicol. 2: 69-78.

 —. 1982b. Relative tolerance of selected aquatic oligochaetes to individual pollutants and environmental factors.
 Aquat. Toxicol. 2: 47-67.

 CHAPMAN P. M., E. A. POWER, R. N. DEXTER, AND H. B. ANDERSEN. 1991. Evaluation of effects associated
        with an oil platform, using the sediment quality triad. Environ. Toxicol. Chem. 10: 407-424.

CHRISTENSEN B. 1984. Asexual propagation and reproductive strategies in aquatic oligochaeta. Hydrobiologia 115:
        91-95.                                                        •?

 CONNELL D. W., AND R. D. MARKWELL. 1990. Bioaccumulation in the soil to earthworm system. Chemosphere
        20:91-100.                                       .

 CONNELL D. W., M. BOWMAN, AND D. W. HAWKER. 1988a. Bioconcentration of chlorinated hydrocarbons
        from sediment by oligochaetes. Ecotox. Environ. Safety 16: 293-302.

 DREWES C. D., AND R. O. BRINKHURST. 1990. Giant nerve fibers and rapid escape reflexes in newly hatched
        aquatic oligochaetes, Lumbriculus  variegatus (family Lumbriculidae). Invertebrate Reproduction
        and Development 17: 91-95.
                                               530

-------
EISENREICH S. J.,  P. D. CAPEL, J. A. ROBBINS, AND R. BOURBONNIERE. 1989. Accumulation .and
       diagenesis of chlorinated hydrocarbons in lacustrine sediments. Environ. Sci. TechnoL 23: 1116-
       1126, ••  '    .    •           .   .   s:\  ..  •  '  '-.:.-                 i    .        .,   '.       . .

FERRARO S. P.* AND F. A. COLE. 1990. Taxonomic level and sample size sufficient for assessing pollution
       impacts on the Southern California Bight macrobenthos. Mar. Ecol. Prog. Ser. 67: 251-262.

FISHER J. B., AND Gf. MATISOFF. 198 1. High resolution vertical profiles of pH in recent sediments. Hydrobiologia
       79:277-284.

FISHER J. B., W. J. LICK, P. L. MCCALL, AND J. A. ROBBINS, 1980. Vertical mixing of, lake sediments by
       tubificid oligochaetes. Journal of Geophysical Research 85: 3997-4006.

FLEMING T.  P., M. K. PRATTEN, AND K. S. RICHARDS.  1982a.  Subcellular localization of zinc hi
       experimentally polluted Tubifex tubifex. Comp. Biochem. PhysioL 73C; 187-193.

FRIES C. R., AND R. F. LEE. 1984. Pollutant effects on the mixed function oxygenase (MFO) and reproductive
               nf the marine polvchaete Nereis virens. Mar. Biol. 79; 187-193.
 FUKUHARA H., AND M. SAKAMOTO. 1987. Enhancement of inorganic nitrogen and phosphate release from lake
        sediment by tubificid worms and chironomid larvae. Oikos 48: 312-320.

 FUKUHARA H., AND K. YASUDA. 1989. Ammonium excretion by some freshwater zoobenthos from a eutrbphic
        lake. Hydrobiologia 173: 1-8.

 GABRIC A. J., D. W. CONNELL, AND P. R. F. BELL. 1990. A kinetic model for bioconcentration of lipophilic
        compounds by oligocbaetes. Water Res. 24: 1225-1231.    -

 GARDNER W. S., T. F. NALEPA, D. R. SLAVENS, AND G. A. LAIRD. 1983. Patterns and rates of nitrogen
        release by benthic chironomidae and oligochaeta. Can. J. Fish AquaL ScL 40: 259-266.

 GIESY J. P., AND R. A. HOKE. 1989. Freshwater sediment toxicity bioassessmenc  rationale for species selection
        and test design. J. Great Lakes Res. 15: 539-569.

 GOERKE H.,  AND K. WEBER. 1990. Population-dependent elimination of various polychlorinated biphenyls in,
        Nereis diversicolor (Polychaeta). Mar. Environ. Res. 29: 205-226.

 GRANEY R.  L., T. J. KE3LTY, AND J. P. GIESY. 1986. Free.amino acid pools of five species .of freshwater
        oligochaetes. Can. J. Fish Aquat. ScL 43: 600-607.

 GRANT A., J. G. HATELEY, AND N. V. JONES. 1989. Mapping the ecological impact of heavy metals on the
        estuarine polychaete Nereis diversicolor using inherited metal tolerance. Mar. Pollut Bull. 20: 235-
        238.             .                          .     .

 GUNN A. M., D. T. E. HUNT, AND D. A. WINNARD. 1989. The effect of heavy metal speciation in sediment
         on bioavailability to tubificid worms. Hydrobiologia 188/189: 487-496.                '        ,

 •HORNIG C. E. 1980. Use of aquatic oligochaete, Lumbriculus variegatus, for effluent biompnitoring, p. 1-9. In
         Industrial Environmental Research Brief,, US EPA-600/D-80-006.

 JAFVERT C. T.  1990. Sorption of organic acid compounds to sediments: „ mitial model development Environ.
         Toxicol. Chem. 9: 1259-1268.
                                                  531

-------
JOHNSON M. G., O. C. MCNEIL, AND S. E. GEORGE. 1987. Benthic macroinvertefeate associations in relation
        to environmental factors in Georgian Bay. J. Great Lakes Res. 13: 310-327.

KAISER M., U. IRMER, AND K. WEILER. 1989. Monitoring of water quality: seasonal variations of heavy metals
        in sediment, suspended particulate matter and tubificids of the Elbe River. Environ. Technol.
        Letters 10: 845-854.

KARICKHOEF S. W., AND K. R. MORRIS. 1985. Impact of tubificid oligochaetes on pollutant transport in bottom
        sediments. Environ. ScL TechnoL 19: 51-56.

KEILTY T. J., AND P. F. LANDRUM. 1990. Population-specific toxicity responses by the freshwater oligochaete,
        Stvlodrilus heringianus. in natural Lake Michigan sediments. Environ. ToxicoL Chem. 9: 1147-
        1154.

KEILTY T. J., AND G. R. STEHLY. 1989. Preliminary investigation of protein utilization by an aquatic earthworm
        in response to sublethal stress. Bull. Environ. Contain. ToxicoL 43: 350-354.

KEILTY T. J., D. S. WHITE, AND P. F. LANDRUM. 1988a. Short-term lethality and sediment avoidance assays
        with  endrin-contaminated sediment and two oligochaetes from Lake Michigan. Arch. Environ.
        Contain. ToxicoL 17: 95-101.

—.' 1988b. Sublethal responses to endrin in sediment by Limnodrilus faoffmeisteri (Tubificidae), and in mixed culture
        with Stylodrilus heringianus (Lumbriculidae). Aquat. ToxicoL 13: 227-250.

KLERKS P. L., AND P. R. BARTHOLOMEW. 1991. Cadmium accumulation and detoxification in a Cd-resistant
        population of the oligochaete Limnodrilus hoffmeisteri. AquaL ToxicoL 19: 97-112.

KNEZOVICH J. P., F. L. HARRISON, AND R. G. WILHELM. 1987. The bioavailability of sediment-sorbed organic
        chemicals:  a review. Water, Air, Soil Pollut 32: 233-245.

KOERTING-WALKER C., AND J. D. BUCK. 1989. The effect of bacteria and bioturbation by Clymenella torquata
        on oil removal from sediment Water, Air, Soil Pollut. 43: 413-424.

KREZOSKI J. R., AND J. A. ROBBINS. 1985. Vertical distribution of feeding and particle-selective transport of
        137Cs in lake sediments by Lumbriculid Oligochaetes. Journal of Geophysical Research 90:11,999-
    .    12,006.                                              .

KRIEGER K. A. 1984.  Benthic macroinvertebrates as indicators of environmental degradation in the southern
        nearshore zone of the central basin of Lake Erie. J. Great Lakes Res. 10: 197-209.

LAKE J. L., N. I. RUBINSTEIN, H. LEE IL C. A. LAKE, J. HELTSHE, AND S. PAVIGNANO. 1990. Equilibrium
        partitioning and bioaccumulation of sediment-associated contaminants by infaimai  organisms.
        Environ. ToxicoL Chem. 9: 1095-1106.

LARSON R. A., AND M. R. BERENBAUM. 1988. Environmental phototoxicity: solar ultraviolet radiation affects
        the toxicity of natural and man-made chemicals. Environ. Sci. TechnoL 22: 354-360.

LAURTTSEN D. D., S. C. MOZLEY, AND D. S. WHITE. 1985. Distribution of oligochaetes in Lake Michigan and
        comments on their use as indices of pollution. J. Great Lakes Res. 11: 67-76.

LAWRENCE T. M., AND T. L. HARRIS. 1979. A quantitative method for ranking the water quality tolerances of
        benthic species. Hydrobiologia 64: 193-196.
                                                532

-------
LEE R. R, S. C. SINGER, AND D. S. PAGE. 1981. Responses of cytochrome P-450 systems in marine crab and
       polychaetes to organic pollutants. Aquat. Toxicol. 1: 355^365.

LODEN M. S. 1974. Predation by chironomid (Diptera) larvae on oligochaetes. Limnol. Oceanpgr. 19:156-159.

LONG E. R., M. F. BUCHMAN, S. M. BAY, R. J. BRETELER, R. S. CARR, P. M. CHAPMAN, J. E.;HOSE, A.
       L. LISSNER, J. SCOTT, AND D. A. WOLFE. 1990. Comparative evaluation of five toxicity tests
       with sediments fromSan Francisco Bay and Tomales Bay, California. Environ. Toxicol. Chem. 9:
       1193-1214.                                                        [ .'.-.'.

MAC M. J., G. E. NOGUCHI, R. J. HESSELBERG, C. C. EDSALL, J. A. SHOESMTTH, AND J. D. BOWKER.
        1990a. A bioaccumulation bioassay for freshwater sediments. Environ. Toxicol. Chem. 9: 1405-
      .  1414. . - '     .'..-:

MARKWELL R. D., D. W. CONNELL, AND A. J. GABRIC. 1989. Bioaccumulation of lipophilic compounds from
       sediments by oligochaetes. Water Res. 23: 1443-1450.  .

MATISOFF G., J. B. FISHER, AND S.  MATIS. 1985. Effects of benthic macroinvertebrates on the exchange of
        solutes between sediments and freshwater. Hydrobiologia 122:  19-33.

MCELROY A. E.  1990. Polycyclic aromatic hydrocarbon metabolism in the polycbaete Nereis virens. AquaL
        Toxicol. 18:35-50.            ,                               ;

MCMURTRY M. J. 1984a. Avoidance of sublethal doses of cooper and zinc by tubificid oligocbaetes. J. Great Lakes
        Res. 10: 267-272.

MOORE D. W., T. M. DILLON, AND B. C. SUEDEL. 1991. Chronic toxicity of tributyltm to the marine polychaete
        worm. Neanthes arenaceodentata. Aquat Toxicol. 21; 181-198.

NEBEKER A. V.,  W L. GRIFHS, C. M. WISE, E.  HOPKINS, AND J. A. BARBTITA. 1989a. Survival,
        reproduction and bioconcentration in invertebrates andfish exposed to hexachlorobenzene. Environ.
        Toxicol. Chem. 8: 601*611.

 NEBEKER A. V., G. S. SCHUYTEMA, W. L. GRIFFIS, J. A. BARBITTA, AND L. A. CAREY. 1989b. Effect
        of sediment organic carbon on survival of Hvalella azteca exposed to .DDT and Endrin. Environ.
      • Toxicol. Chem. 8: 705-718.

 NEFF J. M., B. W.  CORNABY, R. M. VAGA, T. C. GULBRANSEN, J. A. SCANLON, AND D. J. BEAN. 1988a.
        An Evaluation of the Screening  Level Concentration Approach for Validation of Sediment Quality
        Criteria for Freshwater and Saltwater Ecosystems, p. 115-127. In W. J. Adams, G. A. Chapman,
         and W. E.  Landis [eds.], Aquatic Toxicology and Hazard Assessment, ASTM STP 971, American
         Society of Testing and Materials.

 —. 1988b. An evaluation of the screening level concentration approach for validation of sediment quality criteria
         for freshwater and saltwater ecosystems. Aquat ToxicpL 10: 115-127.

 OLIVER B. G. 1984. Uptake of chlorinated organics from anthropogenically contaminated sediments by oligochaete
       -  worms. Can. J. Fish Aquat Sci. 41: 878-883.   •

 --. 1987. Bipuptake of chlorinated hydrocarbons from laboratory-spiked and field sediments by oligochaete worms.
         Environ. Sci. Technol. 21: 785-790.                                                        ;
                                                      533

-------
OLLA B. L., V. B. ESTELLE, R. C. SWARTZ, G. BRAUN, AND A. L. STUDHOLME. 1988. Responses of
       polychaetes to cadmium-contaminated sediment: comparison of uptake and behavior. Environ.
       Toxicol. Chem. 7: 587-592.

PAYNE B. S., AND A. C. MILLER,  1991. The  structure  of dominant invertebrate assemblages in a small
       southeastern stream. J. 6: 257-266.

PESCH C. E., AND P. S. SCHAUER. 1988. Flow-through culture techniques  for Neanthes arenaceodentata
       (Annelida:Polychaeta), including influence of diet on growth and survival. Environ. Toxicol. Chem.
       7: 961-968.

PESCH C. E., W. R. MUNNS JR., AND R. GUTJAHR-GOBELL. 1991. Effects of a contaminated sediment on life
       history traits and population growth rate of Neanthes arenaceodentata (PolychaetatNarftidap.) in the
       laboratory. Environ. ToxicoL Chem. 10:  805-815.

PESCH O. E. 1979. Influence of three sediment types on copper toxicity to the polvchaete Neanthes arenaceodentata.
       Mar. Biol. 52: 237-245.

PRJDMORE R-, S. THRUSH, V. CUMMINGS,  AND J. HEWITT. 1992. Effect of the organochlorine pesticide
       technical chlordane on intertidal macrofauna. Mar. PolluL Bull. 24: 98-102.

REYNOLDSON T. B., D. W. SCHLOESSER, AND B. A. MANNY. 1989. Development of a benthic invertebrate
       objective for mesotropbic Great Lakes waters. J. Great Lakes Res. 15: 669-686.

REYNOLDSON T. B., S. P. THOMPSON, AND J. L. BAMSEY.  1990. A sediment bioassay using the tubificid
       oligochaete worm Tubifex tubifex. Environ. Toxicol. Chem. (submitted).

ROBBINS J. A. 1986. A model for particle-selective transport of tracers hi sediments with conveyor belt deposit
       feeders. Journal of Geophysical  Research 91: 8542-8558.

ROBBINS J. A., T. KEILTY,  D. S. WHITE, AND D. N. EDGINGTON.  1989a. Relationship among tubificid
       abundance, sediment composition, and accumulation rates in Lake Erie. Can. J. Fish AquaL Sci.
       46:223-231.

SAGER M., AND R. PUCSKO. 1991. Trace element concentrations of oligochaetes and relations to sediment
       characteristics in the reservoir at Altenworth/Austria. Hydrobiologia 226: 39-49.

SAMANT H. S., K. G. DOE, AND O. C. VAJDYA. 1990. An integrated chemical and biological study of the
       bioavailability of metals hi sediments from two contaminated harbours in New Brunswick, Canada.
       The Science of the Total Environment 96: 253-268.

SCHUYTEMA G. S. D. F. KRAWCZYK,  W. L. GRIFFIS, A. V. NEBEKER, M. L. ROBIDEAUX, B. J.
       BROWNAWELL, AND J. C. WESTALL. 1988. Comparative uptake of hexachlorobenzene by
       fathead minnows, amphipods and oligochaete worms from water and sediment Environ. Toxicol.
       Chem. 7: 1035-1045.

SCHUYTEMA G. S., D. F. KRAWCZYK, W. L. GRIFFIS, A. V. NEBEKER, AND M. L. ROBIDEAUX. 1990.
       Hexachlorobenzene uptake  by  fathead minnows   and  macroinvertebrates in recirculating
       sediment/water systems. Arch. Environ. Contain. Toxicol. 19: 1-9.

SEDLMEIER U. A., AND K. H. HOFFMANN. 1989. Integumentary uptake of short-chain carboxylic acids by two
       freshwater oligochaetes, Tubifex tubifex and Lumbriculus variegatus:  Specificity of uptake and
       characterization of transport carrier. J. Exp. ZooL 250: 128-134.
                                              • 534

-------
SHAW G. R., AND D. W. CONNELL. 1984. Physicochemical properties controlling polycMorinated biphenyl (PCB)
       concentrations in aquatic organisms. Environ. Sci. Technol. 18: 18-23.

SMITH D. P., J. H. KENNEDY, AND K. L. DICKSON. 1991. An evaluation of a naidid oligocbaete as a toxicity
       test organism. Environ. Toxicol. Chem. 10: 1459-1465.

SOSTER F.  M., AND P. L., MCCALL.  1990. Benthos  response to disturbance  in western Lake Erie:  field
       experiments. Can. J. Fish AquaL Sci. 47: 1970-1985.

STAFFORD  E. A., AND S. P. MCGRATH. 1986. The use of acid insoluble residue to correct for the presence of
        soil-derived metals in the gut of earthworms used as bio-indicator organisms. Environ. PolluL (A)
    ,    42: 233-246.

VAN HATTUM B., K. R. TIMMERMANS, AND H. A. COVERS. 1991. Abiotic arid biotic factors influencing in
        situ  trace metal levels in macroinvertebrates in freshwater ecosystems. Environ. ToxicoL Chem.
        10:275-292.                                     '   ."                                 '

VOGT N. B. 1989. Polynomial  principal component regression: an approach to  analysis and interpretation of
        complex mixture relationships in multivariate environmental data. Chemometrics and intelligent
        laboratory systems. 7: 119-130.

WEBBER E C, D. R. BAYNE, AND W. C. SEESOCK. 1989. Macroinvertebrate communities in Wheeler Reservoir
        (Alabama) tributaries after prolonged exposure to DDT contamination. Hydrobiologia 183: 141-

     •   155-   '        •- •       •                       '.                        *>  *  •
WEDERHOLM T.,.AND G. DAVE. 1989a. Toxicity of metal polluted sediments to Daphnia magna and Tubifex
        tubifex. Hydrobiologia 176/177; 411-417.

 WEDERHOLM T., A. WEDERHOLM, AND G. MILBRINK. 1987a. Bulk sediment bioassays with five species
        of fresh-water oligochaetes. Water, Air,  Soil PolluL 36: 131-154.
                                                   535

-------

-------
      APPENDIX E:
WORKSHOP PARTICIPANTS
           537

-------

-------
Name:       Ray W. Alden
Address:      Old Dominion University
             Applied Marine Research Lab.
             1034 W.45th Street
          •   Norfolk, VA 23529-0456
Phone:       804-683-4195
Fax:         804-683-5293
Name:       Gary Ankley
Address:      ERL-US EPA
             6201 Congdon Blvd.
             Duluth, MN 55804
Phone:       218-720-5603
Fax:         218-720-5539
 Name:       Tom Armitage
 Address:     US EPA/OST
             401 M Street, SW
             Washington, D.C. 20460
 Phone:      202-260-5388
 Fax:        202-260-9830
 Name:       Ray Arnold
 Address:     Exxon Biomedical Sciences
             Mettlers Road
             East Millstone, NJ  08875-2350
 Phone:      908-873-6305
 Fax:        908-873-6009
 Name:       Tom Bailey
 Address:     US EPA
              401  M Street, SW
              Washington, D.C.  20460
 Phone:       703-305-6666
 Fax:         703-305-6309
 Name:
 Address:
 Phone:
 Fax:
Bev Baker
US EPA
401 M Street, SW
Washington, D.C.
202-260-7037
202-260-9830
                               20460
  Name:       Celeste Philbrick Ban-
  Address:     US EPA Region I Laboratory
         '.     60 Westview St.
              Lexington, MA 02173
  Phone:      617-860-4612
  Fax:        617-860-4397
  Name:
  Address:
  Phone:
  Fax:
 Walter Berry
 Sciences Applications International Cor
 c/o  US EPA    ,
 27 Tarzwell Dr.
 Narragansett, RI  02882
 401-782-3101
 401-782-3030
  Name:       G. Allen Burton
  Address:     Biological Sciences Dept.
               Wright State University
               Dayton, OH 45435
  Phone:       513-873-2201
  Fax:         513-873-3301
  Name:
  Address:
   Phone:
   Fax:
 Carrie Buswell
 Viar&Co
 Environmental Services Division
 300 N. Lee Street, Suite 200
 Alexandria, VA  22314-2695
                                                   539

-------
 Name:       Peter Chapman
 Address:     E.V.S. Consultants
             195 Pemberton Avenue
             North Vancouver, EC
             CANADA V7P2R4
 Phone:       604-986-4331
 Fax:        604-662-8548
                                       Name:       Thomas Chase
                                       Address:     EPA/OWOW
                                                   401 M Street, SW
                                                   Washington, D.C. 20460
                                       Phone:       202-260-1909
                                       Fax:         202-260-6294
 Name:
 Address:
 Phone:
 Fax:
Ted Coopwood
EPA/OWEC
401 M Street, SW
Washington, D.C. 20460
202-260-8327
Name:     .  Pat Cotter
Address:     US EPA Region K
            75 Hawthorne Street
            San Francisco, CA  94105
Phone:       415-744-1163
Fax:        415-744-1072
Name:       Philip Crocker
Address:     US EPA-Region VI
             1445 Ross Avenue, 12th fir., Suite 1200
             Dallas, TX  75202-2733
Phone:       214-655-7145
Fax:         214-655-7145
                                      Name:      Joe Cummins
                                      Address:     EPA Region X
                                                  Manchester Laboratory
                                          ,        7411 Beach Drive East
                                                  Port Orchard, WA 98366
                                      Phone:      206-871-8708
                                      Fax:        206-895-4357
Name:       Kristin Day                             Name:
Address:     National Water Research Institute          Address:
             Rivers Research Branch
             867 Lakeshore Rd., PO Box 5050
             Burlington, Ontario, Canada L7R4A6      Phone:
Phone:       416-336-4659                           Fax:
Fax:         416-336-4972
                                                  Ted Dewitt
                                                  AScI
                                                  2111 S. Marine Science Drive
                                                  Newport, OR 97365
                                                  503-867-5030
                                                  503-867-4049
Name:      Robert Donaghy
Address:     ESD/Region m-Wheelirig
            303 Methodist Bldg.
            Wheeling, WV 26003
Phone:      304-234-0241
Fax:        304-234-0260
                                      Name:       Steve Ells
                                      Address:     US EPA/Technical Oversight Section
                                                  401 M Street, SW
                                                  Washington, D.C. 20460
                                      Phone:       202-260-9803
                                      Fax:        202-260-3106
                                                  540

-------
Name:       Dorothy England
Address:     ABC Laboratories, Inc.
            7200 East ABC Lane
            Columbia, MO  65205
Phone:      314-474-8579
Fax:        314-443-9089
Name:       Gary Evereklian
Address:     Microbics Co.
            11908 CanfieldRd,
            Potomac, MP  20854
Phone:      301-762-3793
Fax:        301-762-2534
Name:       Jim Ferretti
Address:     ESD/AMS-Region E
             2890 Woodbridge Ave.
             Edison, NJ  08837
Phone:       908-321-6728'
Fax:         908-321-6616
Name:
Address:
 Phone:
 Fax:
Catherine Fox
US EPA
401 M Street, SW
Washington, D.C.
202-260-1327
202-260-9830
                              20460
 Name:       Howard Fribush
 Address:     US EPA/Analytical Operations Branch
             401 M Street, SW
             Washington, D.C.  20460
 Phone:      202-260-2239
 Fax:        202-260-0524
 Name:       John Giesy
 Address:     Fisheries and Wildlife
             Michigan State University
             East Lansing, MI  48824-1222
 Phone:      517-353-2000
 Fax:        517-336-1699
 Name:       David. Gorodetzky
 Address:     Abt Associates Inc.
              Hampden Square-Suite 600
              4800 Montgomery Lane
              Bethesda, MD  20814
 Phone:       301-913-0500
 Fax:         301-652-3618
 Name:       Carol Haley
 Address:     FDA/CVM
              7500 Standish Place
              Rockvffle, MD  20855
 Phone:       301-295-8695
 Fax:         301-295-8297
  Name:       Dave Hansen
  Address:     ERL-ORD
              27 Tarzwell Drive
              Narrangansett, RI 02882-1154
  Phone:      401-782-3027
  Fax:        401-782-3030
  Name:       MikeHarrass
  Address:     AMOCO Corporation
              200 E. Randolph Drive
              Chicago, IL 60601-7125
  Phone:   .   312-856-5116
  Fax:        312-856-7584
                                                   541

-------
 Name:
 Address:
 Phone:
 Fax:
 Robert Hoke
 SAIC
 411 Hackensack Ave.
 Hackensack, NJ 07061
 Name:      Katherine Hollar
 Address: .    US EPA-Region VI
             1445 Ross Ave., 12th Fir., Suite 1200
             Dallas, TX 75202-2733
 Phone:      214-655-6680
 Fax:        214-655-6490
 Name:       Chris Ingersoll
 Address:      US Fish & Wildlife Service
              4200 New Haven Rd.
              Columbia, MO  65201
 Phone:       314-875-5399
 Fax:         314-876-1896
                                        Name:       Peter A. Jones
                                        Address:     Bureau of Water Quality Mgt.
                                                    50 Wolf Rd.
                                                    Albany, NY  12233  .
                                        Phone:    .   518-457-3651
                                        Fax:      •   518-485-7786     '
 Name:       Darryl Keith
 Address:     US FJPA/ERL-Narragansett
             27 TarzweU Rd.
             Narragansett, RI  02882-1154
 Phone:       401-782-3135
 Fax:         401-782-3030
                                       Name:       Mike Kravitz
                                       Address:     US EPA/OST
                                                    401 M Street, SW
                                                    Washington, D.C.  20460
                                       Phone:       202-260-8085
                                       Fax:         202-260-9830
Name:
Address:
Phone:
Fax:
Jody Kubitz
Michigan State University
Institute of Water Research
334 Natural Resources Bldg.
East Lansing, ME 48824-1222
517-353-9174
513-336-1699
Name:       Janet Lamberson
Address:     ERL-Pacific
             Hatfield Marine Science Center
             2111 S.E. Marine Science Drive
             Newport, OR  97365-5260
Phone:       503-867-4043
Fax:         503-867-4049
Name:       Peter Landrum                          Name:
Address:      Great Lakes Environmental Research Lab   Address:
             2205 Commonwealth Blvd.
             Ann Arbor, MI 48105
Phone:       313-668-2276                           Phone-
Fax:         313-668-2055                           Fax-
                                                   Jim Lazorchak
                                                   EMSL/Newtown Facility
                                                   3411 Church St.
                                                   Cincinnati, OH 45244
                                                   513-533-8114
                                                   513-533-8181
                                             542

-------
Name:       Alex Lechich
Address:     US EPA - Region H
            26 Federal Plaza   '
            New York, NY 10278
Phone:     '212-264-1302
Fax:        212-264-4690
                                     Name:       Henry Lee
                                     Address:      ERL-Pacific
                                                  Hatfield Marine Science Center
                                                  2111 S.E. Marine Science Drive
                                                  Newport,  OR  97365-5260
                                     Phone:       503-867-4042
                                     Fax:         503-867-4049
Name:       Charlie MacPherson  ,
Address:     Tetra Tech Inc.
             10306 Eaton Place, Suite 340
             Fairfax, VA 22030
Phone:       703-385-6000
Fax:         703-385-6007
                                      Name:
                                      Address:
                                      Phone:
                                      Fax:
            John Malek
            US EPA-Region X
            1200 Sixth Ave.
            Seattle, WA 98101
            206-442-1286
 Name:       Victor McFarland
 Address:     US Army Corps of Engineers
             Waterways Experiment, Station
    ,         3909 Halls Ferry Road
             Vicksburg, MS  39180
 Phone:       601-637-3721
 Fax:         601-634-3120
                                      Name:
                                      Address:
                                      Phone:
                                      Fax:
            BethMcGee
            US EPA
            401 M Street, SW
            Washington,  D.C. 20460
 Name:
 Address:
 Phone:
 Fax:
Suzy McKinney
US EPA-Region VI
1445 Ross Ave., 12th fl., Suite 1200
Dallas, TX -75202-2733
Name:
Address:
                                       Phone:
                                       Fax:
Brian Melzian
US EPA-Region DC
c/o ERL-Narragansett
27 Tarzwell Drive
Narragansett, RI 02882
 Name:       Ossi Meyn
 Address:     US EPA/OPPTS
              401 M Street, SW
              Washington, D.C.  20460
 Phone:       202-260-1264
 Fax:         202-260-1283
                                       Name:       William Nelson        .
                                       Address:     US EPA/ERL-Narragansett
                                                   27 Tarzwell Drive
                                                   Narragansett, RI  02882-1154
                                       Phone:       401-782-3053
                                       Fax:        401-782-3030
                                                  543

-------
 Name:       Teresa Norberg-King
 Address:     ORD-ERL-Duluth
              6201 Congdon Blvd.
              DuIuth.MN  55804
 Phone:    '   218-720-5529
 Fax:         218-720-5539
                                       Name:       Warren Norwood
                                       Address:     Dept of Fisheries and Oceans
                                                    867 Lakeshore Rd., PO Box 5050
                                                    Burlington, Ontario, Canada L7R4A6
                                       Phone:       416-336-4694
                                       Fax:         416-336-6437
 Name:       Joe Olha
 Address:     US Army Coips of Engineers
             26 Federal. Plaza
             New York, NY 10278
 Phone:       212-264-5621
 Fax:        212-264-4260
                                       Name:      Jim Oris
                                       Address:     Dept. of Zoology
                                                   Miami University
                                                   Oxford, OH 45056
                                       Phone:      513-529-3194
                                       Fax:        513-529-6900
 Name:
 Address:
Phone:
Fax:
Bill Peltier
ESD
College Station Road
Athens, GA  30613
706-546-2296
Name:      Harriet L. Phelps
Address:     University of the District of Columbia
            Biology Department
            4200 Connecticut Ave., NW
            Washington, D.C.  20008         :
Phone:      202-282-7364
Fax:        301-345-6017
Name:       Michele Redmond
Address:     AScI Corporation
             2111 S.E. Marine Science Dr.
             Newport, OR 97365-5260
Phone:       503-867-5031
Fax:         503-867-4049
                                       Name:
                                       Address:
                                       Phone:
                                       Fax:
            Mary Reiley
            US EPA
            401 M Street, SW
            Washington, D.C. 20460
            202-260-9456
Name:       John H. Rodgers, Jr.                     Name:
Address:     Biological Field Station, Dept. of Biology    Address:
             University of Mississippi
             Oxford, MS  38677
Phone:       601-232-5479                            Phone-
Fax:         601-232-5144                            Fax-
                                                   Lisa Rosman
                                                   US Army Corps of Engineers
                                                   26 Federal Plaza
                                                   New York, NY  10278
                                                   212-264-5621
                                                   212-264-4260
                                                     544

-------
Name:        Norm Rubinstein
Address:      ERL-US EPA
             27 Tarzwell Drive
             Narragansett, RI 02882-1154
Phone:       401-782-3002
Fax:         401-782-3030
                                      Name:       Keith Sappington
                                      Address:     Abt Associates Inc.
                                                   Hamden Square-Suite 600
                                                   4800 Montgomery Lane
                                                   Bethesda, MD 20814
                                      Phone:       301-913-0500
                                      Fax:         301-652-3618
Name:
Address:
Phone:
Fax:
Chris Schlekat
SAIC
165 Dean Knauss Dr.
Narragansett, RI  02882
(401)782-1900
Name:
Address:
Phone:
Fax:
Richard Schwer
Dupont Company
P.O. Box 6090
Newark, DE 19714-6090
Name:       John Scott                              Name:
Address:     Science Applications International Corp.     Address:
             165 Dean Knauss Drive
             Narragansett, RI  02882
Phone:       ,401-782-1900                           Phone:
Fax:         401-782-2330                           Fax:
                                                   Richard Scroggin
                                                   Asticou Centre
                                                   241 Cite Des Jeunes Blvd.
                                                   Hull, Quebec, Canada  K1AQH3
                                                   819-953-5907
                                                   819-997-8427        ...
 Name:
 Address:
 Phone:
 Fax:
Carol Ann Siciliano
US EPA, Office of General Counsel
401 M Street, SW
Washington, D.C. 20460
202-260-8653
Name:
Address:
                                       Phone:
                                       Fax:
Cindy Simbanin
VIAR & Co.
Environmental Services Division
300 N. Lee Street, Suite 200
Alexandria, VA  22314-2695
 Name:       Jerry Smrchek
 Address:     US EPA/OPPT
             401 M Street, SW
             Washington, D.C,  20460
 Phone:       202-260-1268
 Fax:        202-260-1283
                                       Name:       Betsy Southerland
                                       Address:     US EPA  ,
                                                    401 M Street, SW
                                                   .Washington, D.C.  20460
                                       Phone:       202-260-3966
                                       Fax:         202-260-9830
                                                      545

-------
Name:       Mark D. Sprenger
Address:     US EPA/ERT
            2890 Woodbridge Ave., Bldg. 18
            Edison, NJ 08837-3679
Phone:       908-906-6826
Fax:        908-321-6724
                                      Name:       Alexis Steen
                                      Address:     American Petroleum Institute
                                                  1220 L St., NW   .
                                                  Washington, D.C. 20005
                                      Phone:      202-682-8339
                                      Fax:        202-682-8270
Name:
Address:
Phone:
Fax:
William Stelz    •
EPA/ORD
401 M Street,  SW
Washington, DC  20640
202-260-5798
Name:       Eric A. Stem
Address:     US EPA-Region n
            2-WMD, 26 Federal Plaza
            New York, NY  10278
Phone:      212-264-5283
Fax:        212-264-4690
Name:      Margaret Stinson
Address:     Washington Dept. of Ecology
            7411 Beach Dr. East
            Port Orchard, WA  98366
Phone:      206-871-8708
Fax:        206-895-4357
                                      Name:       Elaine Suriano
                                      Address:     US EPA
                                                   401 M Street, SW
                                                   Washington, D.C.  20460
                                      Phone:       202-260-7739
                                      Fax:         202-260-3106
Name:       Irene Suzukida                          Name:
Address:     US EPA                               Address:
             Office of Wastewater Enforcement and
             Compliance
             401 M Street, SW                       Phone:
             Washington, D.C.  20460                Fax:
Phone:       202-260-9536
Fax:         202-260-1460
                                                   Rick Swartz
                                                   ERL-US EPA
                                                   2111 S.E. Marine Science Drive
                                                   Newport, OR 97365-5260
                                                   503-867-4031
 Name:
 Address:
 Phone:
 Fax:
 Bill Telliard
 US EPA/OST
 401 M Street, SW
 Washington, D.C.  20460
 202-260-5131
 Name:       Sharon Thorns
 Address:     Tetra Tech Inc.
             10306 Eaton Place, Suite 340
             Fairfax, VA 22030
 Phone:       703-385-6000
 Fax:        703-385-6007
                                                 546

-------
Name;       Dave A. Tomey
Address:     USEPA-Regiori I
            JFK Federal Bldg.
            Boston, MA 02203
Phone:       617-565-4425
Fax:        617-565-4940
Name:
Address:
Phone:
Fax:
Brian Walls  .
Army Corps of Engineers
Baltimore District
P.O. Box 1715
Baltimore, MD 21203
Name:      Bill Wilber
Address:     USGS/413 National Center
            12201 Sunrise Valley Drive
            Reston,  VA  22092
Phone:      703-648-6878
Fax:        703-648-5295
Name:      Joseph G. Winfield
Address:     Old Dominion University
            Applied Marine Research J-ab.
            1034 W. 45th Street
            Norfolk, VA  23529-0456
Phone:      804-683-4195
Fax::       804-683-5293
Name:      Parley Winger                          Name:
Address::    US FWS/School of Forest Resources       Address:
            University of Georgia
            Athens, GA 30602                      Phone:
Phone:      404-546-2146                           Fax:
Fax:        404-546-2186   "
            BobWischer
            13454 Cotley Lane
            Richmond, VA  23233
            804-780-5202
            804-649-9661
Name:       Douglas A. Wolfe
Address:     NOAA/ORCA
             6001 Executive Blvd.
             Rockville,MD 20852
Phone:       301-443-8933
Fax:         301-231-5764
Name:       Robert Wood
Address:     US EPA
             Office of Wastewater Enforcement
             and Compliance
,             401 M Street, SW
             Washington, D.C.  20460
Phone:       202-260-9536
Fax:         202-260-1460
 Name:       Thomas D. Wright
 Address:     US Army Corps of Engineers
             Waterways Experiment Station
             3909 Halls Ferry Road
             Vicksburg, MS 39180
 Phone:       601-634-3708
 Fax:    .     601-634-3120
Name:
Address:
 Phone:
 Fax:
 Maurice Zeeman
 USEPA/OPPT
 401 M Street, NW
 Washington, D.C.  20460
 202-260-1237
                                                  547

-------

-------

-------

-------