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U.S. Environmental Protection Agency
Region III Information Resource
Center (3PM52)
841 Chestnut Street
Philadelphia, PA 19107
Chesapeake Bay
Ambient Toxicity Assessment
Workshop Report
Workshop held
25-27 July 1989
Annapolis, Maryland
Elizabeth C. Krome
Editor
Prepared by the University of Maryland, the Chesapeake Research Consortium, and
the Chesapeake Bay Research and Monitoring Division
Maryland Department of Natural Resources
May 1990
Printed by the United States Environmental Protection Agency
for the
Chesapeake Bay Program
Printed on recycled paper
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DISCLAIMER
This document has been reviewed by the members of Agency, nor does mention of trade names,
the Ambient Toxicity Assessment Workshop commercial products, commercial or academic
Technical Steering Committee and approved for institutions constitute endorsement or recommendation
publication by the Chesapeake Bay Program, U.S. for use. All responses provided by institutions in
Environmental Protection Agency. Approval does not Appendix C were compiled as presented and are
signify that the contents necessarily reflect the view presumed accurate.
and policies of the U. S. Environmental Protection
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Foreword
The 1983 USEPA report, Chesapeake Bay Program
Technical Studies: A Synthesis, which detailed the
findings of the seven year study of Chesapeake Bay,
found high concentrations of metals and organic com-
pounds in some portions of the Bay, most notably in
highly industrialized areas such as the Patapsco and
Elizabeth Rivers. High levels of metal contamination
were also discovered in sediments in the upper mid-
Bay area, upper Potomac, upper James, and small
sections of the Rappanhannock and York Rivers. In
light of these findings, the link between these toxic
conditions and the impact upon the Bay's living
resources became an important priority in the formu-
lation of the Chesapeake Bay Basinwide Toxics
Reduction Strategy. The authors of the Strategy
recognized that "no critical compendium of scientific
information relating to distribution and effects of
toxics in the Chesapeake Bay has been formulated.
Without such information, developing hypotheses
concerning effects of toxic substances on biota in
Chesapeake Bay is difficult if not impossible". They
also noted that the link between ambient toxicity
assessment and its impact on living resources was not
clear enough to direct appropriate management
actions.
The Ambient Toxicity Assessment Workshop was
convened with the goal of describing the state of the
art in the use of biological indicators. As with any
emerging field, it is clear that dealing with toxics is
highly complex and more research and experience
must be gathered. Creative and innovative methods
need to be tested and existing techniques need further
refinement. Although gaps still remain in our under-
standing of the effects of ambient toxicity on living
resources, enough knowledge has been gained to lay
out many important principles relating whole organ-
ism toxicity testing, sediment toxicity testing, and to
lesser degrees suborganism toxicity testing and
population risk assessment (based on toxicity testing).
This document is a first approach to develop consen-
sus protocols for the use of biological indicators to
monitor the effects of toxic contaminants in Chesa^
peake Bay habitats important to living resources. As
previously stated, the Chesapeake Bay Basinwide
Toxics Reduction Strategy, which was approved and
adopted by the US Environmental Protection Agency,
the Commonwealths of Virginia and Pennsylvania, the
State of Maryland, the District of Columbia, and the
Chesapeake Bay Commission, contains a number of
commitments in the area of research, monitoring, and
toxics management that are necessary to achieve a
comprehensive approach to reduce toxics input to
Chesapeake Bay.
In recognition that research and monitoring programs
will provide new information about the toxics prob-
lem in the Bay, the developers of the Strategy made
the following commitment:
"By July 1989, the signatories agree to con-
vene a scientific workshop to develop con-
sensus protocols for the use of biological
indicators to monitor the effects of toxic
contaminants in Chesapeake Bay habitats
important to living resources."
This workshop was a fulfillment of this commitment.
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ACKNOWLEDGEMENTS
The Ambient Toxicity Assessment Workshop was
conducted by the the University of Maryland and the
Chesapeake Research Consortium, Inc., under
Contract No. CB89-02-023 with the Maryland
Department of Natural Resources as part of the
NOAA Section 309 Interstate Grant project. The
project was conducted under the direction of the 309
Interstate Working Group: Cynthia Stenger, MDNR;
Laura Lower, VA COE; Jerrald Hollowell, SRBC;
David Kaiser, NOAA, OCRM; David Pyoas, Project
Manager, MDNR.
This workshop would not have been possible without
the contributions of a great number of resource
managers, administrators, scientists, and technicians.
Gratitude is also extended to all participants and
supporters, including the workshop's
sponsors, steering and planning committees, speakers,
conveners, workgroup chairs, recorders, attendees, and
those providing financial and logistical support. The
names of contributing individuals appear either below
or in Appendix B.
We would also like to thank Joseph Mihursky and
Karen McDonald of CRC, Inc., who provided
workshop facilitation. Elizabeth Krome provided
technical editing. Many thanks to Stephen Jordan for
his assistance in reviewing the material contained
herein. Layout and graphics were produced through
the patient and careful work of Lamar Platt. The 309
Working Group is particularly grateful to Cindy
Corlett and Pam Owens of CRC Inc., for their tireless
efforts.
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WORKSHOP SPONSORS
Maryland Department of Natural Resources
Virginia Council on the Environment
Susquehanna River Basin Commission
Chesapeake Bay Program Scientific and Technical Advisory Committee
Financial Support
Office of Coastal Resource Management/National
Oceanic and Atmospheric Administration/
U.S. Department of Commerce
Maryland Department of Natural Resources
Logistical Support
Chesapeake Research Consortium
Workshop Steering Committee
Dr. Ray Alden
Applied Marine Research Laboratory
Old Dominion University
Mr. Richard Batiuk
U.S. Environment Protection Agency
Dr. Brian Bradley
University of Maryland
Dr. Katherine Farrell
MD Dept. of the Environment
Dr. Jay Gooch
University of Maryland
Chesapeake Biological Laboratory
Mr. Lenwood Hall
The Johns Hopkins University
Dr. Michael Hirshfield
Maryland Department of Natural Resources
Dr. Karen Hulebak
National Research Council
Dr. Stephen Jordan
Maryland Department of Natural Resources
Dr. Joseph Mihursky
Chesapeake Research Consortium, Inc.
Dr. Jack Plimmer
USDA, Agricultural Research Service
Mr. David Pyoas
Maryland Department of Natural Resources
Mr. Eli Reinharz
Maryland Department of the Environment
Dr. James Sanders
The Academy of Natural Sciences of Philadelphia
Benedict Estuarine Research Laboratory
Dr. Peter Van Veld
College of William and Mary
Virginia Institute of Marine Science
Dr. Dave Wright
University of Maryland
Chesapeake Biological Laboratory
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Workshop Planning Group
Mr. Dan Audet
U.S. Fish and Wildlife Service
Dr. Bette Bauereis
Baltimore Gas & Electric Company
Mr. Richard Batiuk
U. S. Environmental Protection Agency
Mr. Mark Bundy
Maryland Department of Natural Resources
Dr. Arthur Butt
Virginia Water Control Board
Mr. James Hannaham
University of the District of Columbia
Dr. Ian HartweU
Chesapeake Biological Laboratory
Dr. Michael Hirshfield
Maryland Department of Natural Resources
Mr. Jerrald Hollowell
Susquehanna River Basin Commission
Dr. Stephen Jordan
Maryland Department of Natural Resources.
Ms. Laura Lower
Virginia Council on the Environment
Dr. Joseph Mihursky
Chesapeake Research Consortium
Dr. Jack Plimmer
USDA, Agricultural Research Service
Dr. Charles Puffinberger
Maryland Dept. of Agriculture
Mr. David Pyoas
Maryland Department of Natural Resources
Dr. Louis Sage
Academy of Natural Sciences of Philadelphia
Ms. Cynthia Stenger
Maryland Department of Natural Resources
Ms. Debra Trent
Virginia Water Control Board
Others (see Appendix B for full listing of names)
Plenary Speakers
Workgroup Chairs
Conveners
Workgroup Participants
Recorders
Workshop Staff
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TABLE OF CONTENTS
EXECUTIVE SUMMARY iii
INTRODUCTION 1
PLENARY SESSION A:
Population Risk Assessments Based on Toxicity Testing 3
Opening Remarks 3
Assessing Risks to Populations 5
Dr. Donald J. Rodier
Population Risk Assessments Based on Toxicity Testing 13
Dr. Glenn W. Suter
PLENARY SESSION B:
Sediment Toxicity Assays 17
Considerations for Sediment Toxicity Tests 17
John Scott
Contaminated Sediments and Sediment Criteria 23
Dr. Christopher S. Zarba
PLENARY SESSION C
Methodologies for Whole Organism Toxicity Testing 29
Laboratory Testing of Ambient Receiving Waters 29
Steven C. Schimmel
Field Toxicity Testing Procedures 33
Jeffrey Black
PLENARY SESSION D:
Methodologies for Suborganismal (Biochemical and Cellular) Toxicity Testing 41
Rationale for and Relevance of Analysis of Suborganismal Responses
to Contaminant Exposure 41
Dr. D.G. Roesijadi
Techniques for Assessing the Sublethal Effects of Chemical
Contaminants on Aquatic Life 45
Jay W. Gooch
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FINAL PLENARY SESSION
Concluding Remarks and Workgroup Session Reports 53
Opening Comments 53
JosejA Mihursky
Risk Assessment Workgroup Report 53
Ian Hartwell
Whole Organism Workgroup Report > 55
Steve Schimmel
Sediments Workgroup Report 57
Richard Peddicord
General Perspectives on the Role of Biomarkers (Biochemical Measures of Effects)
in the Chesapeake Bay Toxics Workplan 60
Dr. Ken Jenkins, Dr. Brian Bradley, et al.
APPENDIX A: Workgroup Questions and Discussion Notes 63
APPENDIX B: Workshop Participants 85
APPENDIX C: Bioassay Capabilities Survey Results 91
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EXECUTIVE SUMMARY
The Ambient Toxicity Assessment Workshop
provided a forum on how to use biological indica-
tors to monitor the effects of toxic contaminants in
Chesapeake Bay habitats important to living re-
sources.
Why conduct an Ambient Toxicity Assessment
Workshop? Many resource managers must assess
the impacts of toxics on living resources. It is
conceptually attractive to use biological indicators
of actual toxicity, rather than measured levels of
chemical compounds to infer levels of toxicity.
Current bioassessment test protocols, however, often
result in complex and conflicting test results. This
problem prompted the developers of the Chesa-
peake Bay Basinwide Toxics Reduction Strategy to
call for a toxicity assessment workshop. Its pur-
pose was to provide resource managers with the
necessary information to better assess and evaluate
the significance of toxic contaminants as causes of
mortality and impaired growth and reproduction of
Bay organisms.
A three-day workshop of scientists, technicians, and
resource managers from the Chesapeake Bay area
and around the nation met to share knowledge
about the state of the art in toxicity assessment of
aquatic habitats. The workshop consisted of a
series of plenary discussions and task group work-
ing sessions. The first day was a plenary session.
Experts gave presentations in each of four areas of
toxicity assessment:
Population Risk Assessment;
Sediment Toxicity Assessment;
Whole Organism Toxicity Assessment^
Suborganismal Toxicity Assessment.
This initial plenary session involved the presenta-
tion of prepared papers which reviewed the relevant
literature in each major topic and described the
relationship between management needs for infor-
mation and the available methods to fill these
needs.
During days two and three, a selected group of
experts remained to begin synthesizing information
from the initial plenary session. Questions devel-
oped by workshop committees were used to provide
structure, continuity, and commonality among each
of the task groups. Each group, with the exception
of the suborganismal, was charged with developing
its own independent section to include conclusions,
findings, and recommendations on the topic for that
session.
The morning of the third day was used as a wrap-
up session where task group members, within tasks
groups, engaged in a review and a consensus-
building process on the preceding day's products.
The process provided a mechanism whereby discus-
sion and identification of potential problems and
limitations of the research procedures could be re-
solved. The final plenary session was used to
present findings and make recommendations on
how these findings could be applied to a pilot
project.
This workshop resulted in general agreement on
recommendations for a list of suitable methodolo-
iii
-------
gies and species that are appropriate for bioassess-
ment.
Summary of the Opening Plenary Session
The first session was entitled Population Risk Assess-
ment. The presenters were Ian Hartwell (University
of Maryland), Donald Rodier (USEPA, Office of
Toxic Substances), and Glenn Suter (Oak Ridge
National Laboratory). Hartwell provided a historical
perspective on the use of risk assessment. He noted
that risk assessment originated in the insurance and
human health industries, and has only begun to be
applied to ecological and population assessments. As
an environmental aid, risk assessment calculates risk
to populations based on habitat, toxicant chemistry,
and the organism's degree of exposure to a potential
contaminant.
The second session of the opening plenary, Sediment
Toxicity Assessment, worked towards better defining
the complex relationships involving sediments and
contaminants in the aquatic environment. John Scott
(Science Applications International Corporation) and
Chris Zarba (USEPA) presented the current state of
knowledge regarding sediment toxicity testing. Scott
particularly emphasized that sediment toxicity assays
are important because sediments can be long-term
reservoirs of dissolved and particulate-sorbed contami-
nants.
Methodologies for Whole Organism Toxicity Testing
was the title of the third session. Steve Schimmel
(USEPA) and Jeffrey Black (University of Kentucky)
provided an overview of whole organism testing from
both a laboratory and a field testing perspective. This
session evaluated the most accurate, replicable, and
cost-effective methodologies immediately available for
analyzing toxic impacts on biota.
The fourth session, Methodologies for Suborganismal
Toxicity Testing, began with D. G. Roesijadi (Univer-
sity of Maryland) presenting "Rationale for and
Relevance of Analysis of Sub-organismal Responses
to Contaminant Exposure." He suggested that the
effects of toxicity within an aquatic system depend on
both biotic and abiotic characteristics and processes
that undergo complex interactions.
Roesijadi also pointed out that suborganismal respons-
es are generally cellular and subcellular responses.
He stated that one of the difficulties in the study of
suborganismal responses is the problem of relating
information derived at lower levels of biological
organization to more complex levels; in other words,
the ability to extrapolate effects from lower to higher
levels of organization. Jay Gooch (University of
Maryland) emphasized that the purpose of a suborgan-
ismal test is different from one testing the whole
organism. He pointed out that one of the original
justifications for the use of sublethal endpoints in
environmental testing was the desire to have an early
warning system. He went on to describe how suborg-
anismal testing can be used to detect more subtle
effects than traditional endpoints such as death. The
relative merits of these two different approaches were
discussed throughout the workshop.
Summary of Final Plenary Session
The final plenary session addressed the following
concerns:
• identification of acceptable existing methods
for risk assessment, whole organism testing,
and sediment toxicity testing;
• suggestions for modification of existing
protocols; and
• analytical discussion of the species and
protocols identified.
The participants further distinguished between devel-
oped and developing bioassays. The former class is
dominated by acute lethality tests; the latter class is
IV
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dominated by chronic and sublethal effects, some of
which have not yet been field-verified. They also
noted that laboratory studies can establish causal rela-
tionships, but their applicability to the field must be
established. Each recommended bioassay was consid-
ered using the following factors: 1) biological re-
sponses; 2) technology available; 3) cost; 4) sensitivi-
ty to sublethal toxicity; and 5) significant effects.
Population Risk Assessment Group
The risk assessment group recognized the need for a
quantitative approach. However, they indicated that
initially the approach would need to be qualitative
until a sufficient database was developed to allow a
more quantitative assessment. This group recom-
mended a mechanism that would allow researchers to
characterize site-specific results in a ranking scheme
to contrast different sites (Table 1). The ranking
scheme will require additional work to evaluate
precisely how to determine each ranking factor;
however, the group did assign weights to each factor
according to a consensus on its importance in evalu-
ating toxicity.
• Field effects (such as population depressions)
may be due to direct toxicity or indirect
community effects.
This group also made the following recommendations:
• Create databases of the relative sensitivities
of assay species.
• Conduct specific cause-and-effect chemical
studies.
• Estimate chemical exposure either directly or
from historical data if the site is known.
• Define receptors and endpoints after an
assessment of field biomonitoring, sublethal
responses, and/or surrogate species.
• Design research to address a well defined
gradient and a timefactor.
• Develop modeling to link field toxicity
bioassays to population impacts.
T«bU 1. Risk Assessment Ranking System
Ranking Factor Weight
Consistency of results 5.8
Severity of endpoints 8.5
Degree of response 7.9
Number of tests 4.8
Reproducibility of results 6.6
Note: Each member of the group subjectively assigned a score
(1-10) to each factor to reflect its importance. The ranks were
then averaged ID provide an overall indicator of the group's
opinion.
They concluded the following:
• Tests employing more than one species yield
more information but require sensitivity
calibration.
• Address population parameters using end-
points from biomonitoring tests.
Sediment Toxicity Assessment Group
Participants agreed that sediment quality criteria
(SQC) are needed. Because a large database would
be required in order to develop SQC, the group
emphasized the need to collect data supporting
toxicity testing and chemical analysis. Recommen-
dations from the sediment toxicity group were limited
to tests that could identify hot spots and examine the
condition of the sediment as a result of source inputs.
In light of this, they placed strong emphasis on
available methods that would include: (1) the 10-day
amphipod test currently conducted in Puget Sound;
and (2) basic screening tests that have been fairly
cost-effective in Great Lakes (Table 2).
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BIOASSAYS FOR SEDIMENTTOXICITYTESTING IN FRESHWATER, BRACKISH SALTWATER, AND
HIGH-SAUNFTY ENVIRONMENTS IN THE CHESAPEAKE BAY
Organism
Standard bioassays
Freshwater
Hyallela
Chironomus
Hexagenia
Brackish saltwater (0-15 ppt)
Hyallela
Eohaustorius
High salinity
Rhepoxinius
Amoelisca
Bivalve larvae 48-hr
pediveliger oyster
Mercenaria
Polychaete Neanthes (Nereis)
Mysid {grass shrimp)
1
X
X
X
X
X
X
2
X
X
X
?
X
X
X
X
3
X
X
X
X
X
X
X
4
X
X
X
X
X
5
X
X
X
X
X
X
X
6
X
X
X
X
X
X
X
X
7
X
X
X
X
X
X
X
B
X
X
X
X
9
X
X
X
X
X
X
10
So
So
So
So
So
So
SS
So
So
11
a.b
a,b
a,c
a.d
12
L
L
L
L
L
L
L
L
M
M
Notes
Bioassays available based on
enhancement of standard
techniques
Low salinity
Leptocheirus
Eohaustorius
High salinity
Leoidactylus
X
X
X
X
7
Bioassay techniques under develop-
ment or recommended for
development
Chronic tests
Leotocheirus
Lepidactylus
Sago pondweed
X
X
?
?
X
X
X
X
X
X
X
X
X
X
X
X
X
So
So
So
M
M
M
X
So
So
So
H
H
H
(10) So-solid phase; SS-eediment slurry
(11) axacute lethal; b-behavtoral effect; c°abnormal development; d*metMnorphic feJure, etc.
(12) L=tow cost; M.moderats cost; H»hkjh cost
Teble 2. Bioessays for sediment toxicity testing in freshwater, brackish saltwater, and Man-salinity environments in the Chesapeake
Bay.
VI
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Participants suggested the use of biological tests
which use infaunal species to examine benthic com-
munity structure.
The group made the following recommendations:
Use a multifaceted approach to choose the
tests and organisms.
Demonstrate the utility of an amphipod test
for the Chesapeake Bay using existing stan-
dard methods and species indigenous to the
Chesapeake Bay.
Develop and field-validate tests for chronic
and population effects of toxic sediments
using Chesapeake Bay indigenous species.
Whole Organism Toxicity Assessment Group
This group took as its primary charge the question of
what were the most appropriate laboratory and field
toxicity tests for evaluating ambient toxicity in
Chesapeake Bay. To address this issue the group
developed a table of established and accepted meth-
ods, which considered species, duration, and type of
test to be used (Table 3). Several species were rec-
ommended for evaluating ambient toxicity.
The participants emphasized that the choice of test
organisms is contingent upon whether the proposed
study has regulatory endpoints. The group suggested
that species selection should encompass: 1) species
pertinent for regulatory purposes; and 2) species not
pertinent for regulatory purposes. Additionally, they
agreed that no single test was adequate. Instead, they
considered a suite of tests necessary to characterize
toxic effects. Participants strongly recommended
those testing methods that are being used as part of
the Maryland and Virginia NPDES permitting pro-
cess. They also recognized the need to use ecological
indicator species and species amenable to laboratory
testing.
Final Plenary Session Recommendations
Participants in all three sessions of the Ambient
Toxicity Assessment Workshop reached the same
technical consensus on the following issues. These
include:
Toxicity tests which measure growth and
reproduction, as well as survival, are highly
desirable.
Acute lethality tests for Bay species are not
sensitive indicators of ambient toxicity and
should not be used routinely. However, in
areas where high toxicity is suspected, acute
lethality testing may be used as an initial
screen.
Chronic or partially chronic tests with suble-
thal endpoints are the preferred methods.
Test categories were established that includ-
ed: 1) established regulatory methods; 2)
established Chesapeake Bay specific meth-
ods; 3) Chesapeake Bay research methods
still being developed.
Toxicity assessments should recognize the
following major salinity regimes: 1) fresh-
water (0 ppt salinity); 2) estuarine (> 0 to 20
ppt); 3) marine (> 20 ppt).
In summary, information generated by this workshop
will offer resource managers a technical appraisal of
the strengths and weaknesses of methods potentially
available to develop final guidance for a pilot study of
ambient toxicity in Chesapeake Bay. Findings and
conclusions from this pilot study will be used to
develop a Basinwide ambient toxics monitoring
program.
VII
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Table 3- Whole organism tests for ambient toxicity in freshwater, estuarine, and marine
environments of the Chesapeake Bay.
Category
Proven Regulatory
Methods
Established Methods
Pertinent to
Chesapeake Bay
Research Methods
Pertinent to
Chesapeake Bay
Freshwater environment
Ceriodaphnia 7-day chronic
Fathead minnow 7-day chronic and
embryo/larval test
Selenastnim 96-hr
Embryo larval: Muegill, catfish
Striped bass larval toxicity test
Duckweed 96-hr
Estuarine environment (max. 25 pot)
Sheepshead minnow 7-day chronic and
embryo/larval test'
UtnkSa bttySna 7-day chronic
Uysidopsls balUa 7-day chronic
Bivalve larvae:
Crassostreavirginica
Uyaannaria
Mercanaria mtfcenaria
Skefefonema algal test 48 hr?
Grass shrtmp larval acute lethaMy, 96 hr
Striped bass larval toxicity test (tow salinity)
Anchoa mitchl* 96-hr test
Callagtossa algal chronic
Sago pondweed toxicity test
Euiytemora alfinis
Neomysis americana 96-hr acute
Acartiatonsa
Marine environment
Sea urchin fertlizatton test
Utricfa beryiUna 7 -day chronic
Sheepshead minnow 7-day chronic
Uysktopsis bahia 7-day chronic
Bivalve larvae test: Myfflus eduSs
Champia pamila reproductive
Grass shrimp larval acute lethality, 96 hr
Striped bass juvenile 96-hr acute lethality
Attartia tonsa 7-9 days
Anchoa mitchilf 96-hr acute lethality (larval)
Vlll
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INTRODUCTION
This document contains the proceedings from the
Ambient Toxicity Assessment Workshop which was
held in Annapolis, Maryland on July 25-27, 1989.
Funding for the Ambient Toxicity Assessment Work-
shop was provided by the Coastal Zone Management
Act of 1972 (as amended), Section 309 (Interstate
Grants) as administered by the Office of Ocean and
Coastal Resource Management, National Oceanic and
Atmospheric Administration. State logistical and
planning support was provided by the Section 309
Planning Work Group, Maryland Department of
Natural Resources, Virginia Water Control Board,
USEPA Chesapeake Bay Liaison Office, and the
Chesapeake Bay Program Scientific and Technical
Advisory Committee.
The workshop was held in response to two commit-
ments outlined in the Chesapeake Bay Basinwide
Toxics Reduction Strategy (Chesapeake Executive
Council, 1988). One commitment is:
"By July 1989, the signatories agree to convene a
scientific workshop to develop consensus protocols
for the use of biological indicators to monitor the
effects of toxic contaminants in Chesapeake Bay
habitats important to living resources."
The findings and conclusions from this Workshop are
intended to provide guidance for the implementation
of a related commitment:
"By December 1989, the signatories commit to
develop and begin to implement a plan for Baywide
assessment and monitoring of the effects of toxic
substances, within natural habitats, on selected com-
mercially, recreationally and ecologically important
species of living resources."
The report is arranged as follows: first the report of
the opening plenary session, followed by the report of
the final summary session, then by Appendices
containing (A) workgroup session reports, (B) list of
participants, and (C) results of the Bioassay Capabili-
ties Survey.
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Plenary Session A:
Population Risk Assessments Based on Toxicity Testing
Convener: Ian Hartwell
Opening Remarks
I want to put forth a few general comments on what
ecological risk assessment is and how it can be used
before we hear more detailed discussions on numeri-
cal approaches and applications from the plenary
speakers. The basic concepts and approaches to risk
assessment were originally developed in the fields of
insurance and human health for the purposes of
predicting longevity and injury frequency. These
ideas have been expanded and adapted to engineering,
industrial operations, and other fields, and most
recently have been applied to pollution assessments.
Fundamentally, a risk assessment is a calculation of
the probability of some outcome, based upon the
hazard of some input parameter and the degree of
exposure to that parameter. The application of risk
assessment to ecological situations is a relatively new
field. As with any new field, there are a lot of
specialized terms that mean specific things to those
involved in the area of risk assessment, but that are
often used interchangeably by those not working
directly in that field. These inconsistent uses are a
source of confusion. Thus I would like to review
briefly some of the terminology and approaches used
in risk assessment. It is important to realize that
different kinds of assessments yield different products
with specific uses.
"Hazard assessment" is a determination of the inher-
ent danger of a given chemical substance or activity.
It requires a measure of some sort of effects endpoint
and a dose.
It is a quantification of the relationship between dose
and response. It should include information on both
dose concentration and duration of dose. How
inherently dangerous is the material in question,
without regard to potential exposure?
"Exposure assessment" is an estimation of concentra-
tions of a chemical substance to which target organ-
isms are exposed. It includes estimates or measures
of quantity of material, fate and transport modeling,
transformations, and bioconcentration potential. The
estimates are made without regard to the hazard of the
material. Where does it go, and in what concentra-
tion? What are the pathways to the target organisms,
and how long will they be exposed?
"Receptor characterizations" address the entity of
interest in the risk assessment that may be affected by
exposure to chemical substances. In our context, this
may include a species, a population, a community, or
an ecosystem. We need to be able to define what the
receptors are and what other important parameters
affect exposure or response, such as sensitive life
stage, migratory patterns, feeding habits, etc.
"Risk assessment" is the integration of the hazard
assessment, the receptor characterization, and the
exposure assessment. It estimates the magnitude and
probability of harm resulting from exposure of the
target organisms to a hazardous substance or activity.
This term includes all of the other three and is thus
more complex, and it requires that the other three be
compatible.
There are some other items used in combination with
these terms that are relevant here. These are "endan-
germent assessments" and "damage assessments."
"Endangerment assessments" are used in the Super-
fund process for waste site remediation. They are
basically a preliminary, hypothetical risk assessment
for a National Priority List site or a proposed site to
-------
document whether there exists current or potential risk
of significant exposure to toxic chemicals. They
examine the presence of chemicals for dangerous
concentrations, potentially exposed populations, and
the presence of exposure pathways. That is as far as
they go; they do not make predictive conclusions
beyond either "Yes, there is potential risk," or "No,
there is not."
"Damage assessments" are field-based evaluations of
environmental damage after the fact. They were
designed for spill sites. That is, after a release of
toxic chemicals in harmful quantity, the damage
assessment is a means of quantifying the environmen-
tal harm and (in current practice) arriving at a mone-
tary cost for reparations and/or cleanup. That is not
to say that damage assessment procedures could not
be used as the basis for input into a full-blown risk
assessment.
The approaches in generating a risk assessment can be
grouped into qualitative and quantitative methods. In
part, the method of approach is governed by the
proposed uses of the assessment. You can use them
to help set priorities, for example, for disposal op-
tions, for site selection, or for research directions.
You can use them to set standards. You can use
them as input into risk management at the regulatory
level or for resource protection or restoration.
Qualitative methods are obviously useful when you
have little or no data. They rely heavily on profes-
sional judgment and can be used for isetting priorities
as ranking or screening procedures. They are not
useful for standards development or risk management.
They don't yield quantitative estimates of magnitude
or probability of effect.
Quantitative methods can be broadly divided into
quotient methods and exposure-response methods.
Quotient methods basically compare some predicted
effect level with a benchmark standard. This requires
that you have a standard against which to compare
your environmental concern level. The concern level
can be a toxic concentration, an acceptable level of
population reduction, or some other measure of
environmental effect, which may or may not be an
extrapolated value. Quotient methods give you a
yes/no prediction, or in some methods a yes/no/maybe
prediction. They do not give you a probabilistic
result, only an indication whether you are above or
below some benchmark.
Exposure-response methods are modeling approaches
and can be subdivided into top-down or bottom-up
methods. These methods require vastly larger data
sets and knowledge of the relevant environmental
system than other approaches. However, they have
the potential to provide both a prediction of magni-
tude and probability of effect.
Top-down models require ecosystem response data as
input (that is, field data). They evaluate impacts on
the ecosystem directly. They don't provide a mecha-
nistic reason for the changes that are seen or predict-
ed. The models don't need to and cannot determine
the structural or functional changes occurring in the
environment as a result of chemical or physical
effects.
Bottom-up methods require laboratory toxicity data
and a great deal of site-specific and chemical-specific
data. They can model community-level effects, and
through the model development process they can
provide information on how the system works. At
present we still don't have the tools to incorporate all
of the aspects of life history, fate and transport of
chemicals, or stochastic variations in the ecosystem
into unified models, particularly for a system as large
as Chesapeake Bay. However, great progress has
been made over the last several years and new
methods of population compensation modeling are
being developed.
All quantitative methods must also have some means
of estimating uncertainty in parameter estimation. We
-------
need data on confidence intervals for all our input
data and our extrapolations. This can often be one of
the most difficult aspects of risk assessment. How
confident are we that this result will protect the
environment — a factor of two, an order of magni-
tude, three orders of magnitude? This has to be
calculated or estimated. And there are a variety of
approaches such as sensitivity analysis, calibration, or
validation, depending upon what type of risk assess-
ment approach you are using.
Finally I want to say a few words on the difficulty of
applying to the environment a mathematical procedure
whose basic concepts were developed for human
health predictions. We need to be mindful that some
people who will be using the results of our efforts do
not always understand that the databases like those
available for risk assessment in human health simply
do not exist for any other species in such detail. As
Dick Tucker, formerly with the EPA, used to say, we
know how much exposure to benzene will cause
chronic headaches in people, but we will never know
how much it takes to produce the same effect in
ducks. We don't have the databases to achieve the
same precision. There are thousands of species to be
considered, with thousands of interactions and b'fe
history details which we do not know, and there are
myriads of other influences such as habitat integrity,
environmental resilience, and normal ecological
variability which increase our uncertainty. It is
possible to do ecological risk assessments, but the
inputs and the nature of the target populations dictate
that the approach and the results may not be as clean-
cut as health assessments.
This may sound like a pessimistic introduction to the
prospects of ecological risk assessment, but it's im-
portant to remember we are dealing with vastly more
complex systems than risk assessments have ad-
dressed before. Risk assessments are currently being
used successfully in the regulation of pesticides,
effluents, new industrial chemicals, and air pollution,
and they are under development for use at Superfund
sites. Risk assessment provides a scientific basis for
decision-making, for resource allocation, and for
answering questions which concern the regulatory
community. For those reasons, it has great potential
for improving our ability to communicate scientific
concerns that are both politically and legally defensi-
ble to the public, industry, and resource managers.
Assessing Risks to Populations
Donald J. Rodier
Introduction
Good morning and thank you for inviting me to your
workshop. I am a biologist in the Environmental Ef-
fects Branch, which is in the Health and Environ-
mental Review Division of the Office of Toxic
Substances. We are a support branch of the US EPA
and are responsible for providing ecological hazard
and risk assessments of industrial chemicals. Because
these chemicals are commonly discharged to bodies of
water, we have concentrated our efforts on developing
methods for evaluating both the hazard and risk of
these chemicals to aquatic populations.
There are two main categories of industrial chemicals:
1) new chemicals and 2) existing chemicals. Existing
chemicals are those on the Toxic Substances Control
Act (TSCA) inventory (a list of chemicals manufac-
tured before July 1, 1979) and in common use.
Chemicals that are not on the TSCA inventory are
considered new. The two types differ in regard to the
amount of data and the time frames in which we have
to act upon them.
Before proceeding further, it must be emphasized that
while considerable progress has been made in the area
of ecological risk assessment with regard to popula-
tion and ecosystem models, such methods are still in
the developmental stage and have not been used in
regulating industrial chemicals. At this time, we are
not aware of any one fully accepted method that will
translate the data from laboratory assays to precise
-------
predictions of effects on natural aquatic populations.
Conceptual Framework of an Ecological Risk
Assessment (Figure 1)
Ecological risk assessment is a logical process where-
by one integrates the results of the hazard and expo-
sure assessments of a chemical or other stressor into
a statement regarding the probability and consequenc-
es of an adverse ecological effect.
Unlike human health risk assessments, which have
readily understood endpoints such as carcinogenicity,
teratogenicity, etc., ecological risk assessments utilize
endpoints obtained from lexicological assays. These
include an algal EC10, a fish LC50, and Maximum
Acceptable Toxicant Concentration (MATC). Fre-
quently we are asked "What is the real significance if
such endpoints are exceeded in the wild?" That is,
what will happen?
Figure 1.
ECOLOGICAL
HAZARD
ASSESSMENT
ENVIRONMENTAL
EXPOSURE
ASSESSMENT
ECOLOGICAL
RISK
ASSESSMENT
PROBABILITY
OF AN
ADVERSE
EFFECT
QUE
OF AN
ADVERSE
EFFECT
THE ECOLOGICAL
RISK ASSESSMENT PROCESS
of Regulatory Concern." This term was arrived at
after a review of 268 environmental legislative acts.
In summary, all past and present environmental
legislation was enacted to protect natural resources
(both biotic and abiotic) which were valued by soci-
ety, from any reduction, degradation, or loss in any
quality, quantity, or utility (Clements, 1983). After
reviewing nine incidents of how chemicals affected
natural biotic resources, we concluded that chemicals
caused adverse effects on growth and development,
mortality, and reproduction, and that such effects were
manifested at the population level of organization.
Putting all of this together (Figure 2), we see that the
focus of an ecological risk assessment is on natural
populations that are valued by society for various
reasons (aesthetic, commercial, ecological) or are
already protected under different statutes such as the
Endangered Species Protection Act.
Flgur* 2.
POPULATION FAULT TREE
In the Environmental Effects Branch, we relate the
results of ecological risk assessments to "Populations
-------
The three factors that govern a population are growth
and development, mortality, and reproduction. If
individuals fail to grow and develop, the population
may be in danger of not surviving. In addition, if the
population is a commercially valuable one, the utility
of that population is likely to be reduced. If there is
mortality above the normal death rate, the population
may also be in danger. And if individuals within the
population fail to reproduce, the population is obvi-
ously in danger of extinction.
Toxic chemicals can adversely affect mortality,
growth and development, and reproduction in two
ways: direct and indirect. Direct effects are simply
toxic effects with direct impact on growth and devel-
opment, mortality, and reproduction of a specific
species or population. In evaluating the hazard of a
particular substance, we evaluate the direct toxic
effects of a substance to various surrogate species.
Indirect effects are more difficult to define, interpret,
and measure. Indirect effects include disruptions in
the habitat of a particular population and alterations in
the food supply. Natural populations do not exist in
a vacuum. The well-being of one population is
dependent upon other populations which either act as
a food source for that population or play a role in
governing the growth of that population through
predation or grazing. Toxic chemicals can alter these
functions.
In addition to the effects of a toxicant, we know that
unperturbed populations are influenced by their
natural environment, and the term "natural causes" is
a catch-all to include the natural variability associated
with population maintenance. In addressing the risks
posed by toxic chemicals, both the short- and long-
term effects must be considered. Such evaluations are
obtained from the hazard assessment.
I want to emphasize that Figure 2 presents the con-
ceptual framework of an ideal ecological risk assess-
ment -- that is, the ability to quantitatively address the
risks of direct and indirect toxic effects to specific
populations of regulatory concern. We are not there
yet but we are making progress. As of right now, we
can only make inferences to effects on such popula-
tions.
How Ecological Risk Assessments Are Conducted
in EEB (Figure 3)
Existing chemicals. Chemicals are referred to EPA
by an Inter-Agency Testing Committee for testing.
We have one year to respond. Testing proceeds
according to procedures we have set forth (USEPA,
1983). The tests utilize surrogate species (USEPA,
1982) and published guidelines (USEPA, 1985).
Figur* 3.
THE ECOLOGICAL RISK ASSESSMENT PROCESS
L GATHER INFORMATION
A. TOXOCOLOGICAL EFFECTS
B. POTENTIAL EXPOSURE
I. ASSESS THE HAZARD THROUGH TOXKITY
ENOPOWTS
M. ASSESS THE EXPOSURE AND ESTIMATE A
PEC
IV. ASSESS MSK
OUOTCNT METHOD COMMONLY
EMPLOYED
fXPOSUHEfmCTS « OUOTtCNT
AS THE QUOTIENT APPROACHES 1
Oft MORE A HIGH RISK B DEMOTED
AS THE QUOTIENT DECREASES
FROM UNITY LESS OF A RISK B
ASSUMED
V. ASSESS WPACT
VI FORMULATE AND PRESENT THE
CONCLUSIONS
New chemicals. New chemicals, or PMNs (premanu-
facture notice submissions), present certain problems.
The major problems are the large number submitted
(over 1000 annually), the short turn-around time (90
days allowed for evaluation), and the scarcity of data
(USEPA, 1986).
Because there is little data to evaluate the ecological
hazards of a PMN, EEB utilizes structure activity
relationships (SARs) to estimate potential toxicity
(Auer et al. 1989). SAR includes an evaluation of a
chemical by comparison to another chemical for
which there is data (analog) and also through regres-
sion analyses of specific chemical classes.
-------
A large number of these SARs have been compiled
and are available (Clements, 1988). To predict the
concentration of a particular PMN that is likely to
cause some adverse effect in the environment, assess-
ment factors (USEPA, 1984) are used (Figure 4).
Assessment factors are numbers that account for the
uncertainties due to variables such as test species
sensitivity to acute and chronic toxicity, laboratory
test conditions, and age group susceptibility.
Exposure assessments are conducted in the Exposure
Evaluation Division. Models are used to predict the
concentrations of a particular PMN in various stream
reaches (percentiles). In the initial assessment,
estimates are made of concentrations under mean and
low flow conditions. Later, a concern level derived
using assessment factors is used with the probabilistic
dilution model (USEPA, 1988) in order to evaluate
how often (i.e., how many days out of the year) that
concern level will be exceeded.
Flgur«4.
APPLICATION OF ASSESSMENT FACTORS TO
EVALUATE NEED FOR TESTING
DATA AVAILABLE
OSAR-CALCULATED
LC50
SINGLE LC50 FOR
ANALOG
TWO LC50'« FOR PMN
(•.Q., 1 FISH,
1 INVERTEBRATE)
TWO LC50'« FOR SAME
ANALOG («.g. 1
ALGAE, 1 FISH
THREE LC50'« FOR
PMN (FISH, ALGAE
INVERTEBRATE
THREE LCSO's FOR
SAME ANALOG (FISH,
ALGAE, INVERTEBRATE)
FIVE LC50'» FOR PMN
(«.g. 2 FISH, 3
INVERTEBRATES)
FIVE ICSO'm FOR SAME
ANALOG (•.» 3 ALGAE,
2 FISH)
MATC FOR ANALOG
MATC FOR PMN
ASSESSMENT FACTOR TO
BE APPLIED
1000X
1000X
1000X
1000X
100 X
100X
100X
10X
DATA-BASED DECISION ON NEED
FOR FURTHER TESTING OH
REGULATORY DECISION
(ASSESSMENT FACTOR NOT USED]]
The quotient method (Barnthouse et al., 1986) is used
to compare the concern level or lexicological endpoint
with the various exposure concentrations or with the
probabilistic dilution model. This method has been
used successfully both by the Office of Toxic Sub-
stances (Rodier, 1987) and the Office of Pesticide
Programs (Urban and Cook, 1986). However, the
following deficiencies associated with the quotient
method have been noted:
• It does not routinely take into account dose
responses, other than standard lexicological
measurements such as an LC50 or MATC.
• It has no predictive capability.
It does not address taxonomic or life-stage
sensitivities to a toxic chemical.
• Indirect effects of a toxicant are not readily
addressed.
• Implicit in the use of the quotient method is
the assumption that the exposure duration
will be as long as (or longer than ) the
lexicological tesl duralion. Thus pulse or
short-term exposures are not readily address-
ed.
• The uncertainties associaled wilh exlrapola-
lions from laboralory dala lo nalural environ-
ments are nol easily addressed.
-------
Refinements to the Quotient Method
Figure 5.
1 1
•*. OUOTCNT
METHOD
COHMMTM
TOUCOLOOV
«» ANM.TMOF
(XTHAMHATION
UHOM
•on
.*-«
POFUlATOM
•OOCLS
•*. HAH**
tCOSYSTIM
UHCCMTAWTV
AMM.VIW
CURRENTLY AVAILABLE
ECOLOGICAL RISK ASSESSMENT TECHNIQUES
Figure 5 shows some of the extant ecological risk
assessment methodologies. The quotient method has
already been discussed. The author of the Extrapola-
tion of Error method, Dr. Glenn Suter, has previously
discussed this methodology in depth. The Risk
Analysis and Management Alternatives System
(RAMAS) was originally designed for the Electric
Power Research Institute (Rohlf et al., 1986).
RAMAS is a user-friendly Monte Carlo Simulator of
age-structured populations. It is designed to answer
the following: 1) What is the probability that a
population size will fall below a given threshold
within a specified time, and 2) what are the number
of individuals in a certain age class within a specified
time? EEB currently has a rainbow trout version of
RAMAS which is still being evaluated. Inputs to
population models include maximum age of a given
population, fecundity, survivorship among the various
age classes, and estimates of toxicant-induced mortali-
ty among the age classes.
like to briefly explain the concept and then show how
it can be used to address the "Consequences" part of
an ecological risk assessment.The model used in EUA
is the Standard Water Column Model (SWACOM).
SWACOM mimics the aquatic populations that occur
in the water column. There are four groups of
populations represented: (1) phytoplankton or algae
whose biological role is the fixation of energy through
photosynthesis, (2) zooplankton or aquatic inverte-
brates that consume phytoplankton and serve as a
food source for (3) the forage fish, which in turn are
fed upon by (4) the top carnivore or predator fish.
The top predator fish can be thought of as a popula-
tion of regulatory concern.
Flgur* 6.
ECOSYSTEM UNCERTAINTY ANALYSIS
OAK RIDGE NATIONAL LABORATORY
VERSION 3.1 EUA-SWACOM 1987
RISK SUMMARY
CHEMICAL: PHENOL
EXPOSURE (mg/L): 10.000
TROPHIC LEVEL -25% -50% -75% -95% +10%
PHYTOPLANKTON 0.28 0.25 0.11 0.07 0.50
ZOOPLANKTON 0.61 0.51 0.46 0.38 0.29
FORAGE FISH 0.43 0.38 0.33 0.27 0.54
GAMEFISH 0.81 0.77 0.74 0.71 0.16
aoo
!« 200
0 tO 40 «0 «0 100
PCMCCNT REDUCTION M ANNUM. PRODUCTION Of ZOOPLANKTON
The Ecosystem Uncertainty Analysis (EUA) was
developed by O'Neill et al., (1982) at the Oak Ridge
National Laboratory as part of the Office of Research
and Development Synfuel Program. Believing that
the EUA held some promise in addressing indirect
toxic effects, we sponsored work on additional
refinements (Bartell, 1987, 1987a). Time does not
permit discussion of all aspects of EUA, but I would
Figure 6 shows what could happen if a chemical
which was highly toxic to zooplankton entered a lake.
Five simulations with SWACOM were performed
(Bartell et al., 1987). Each simulation decreases the
zooplankton in 20% increments and represents a one-
year cycle in a lake. The decrease in zooplankton
causes a decrease in the forage fish population. The
-------
top carnivore population decreases because of the
declining forage fish population. Because the phyto-
plankton are no longer grazed by the zooplankton,
they begin to increase. This can lead to so-called
algal blooms, which are undesirable not only because
they discolor bodies of water and create noxious
odors but also because they decrease the oxygen
content of the water, causing fish kills. This example
demonstrates the importance of indirect effects, and
the model simulations aid greatly in both evaluating
and explaining such effects.
Since 1986, The Office of Research and Development
has been supporting an ecological risk assessment
initiative for the Office of Pesticides and Toxic
Substances (OPTS). Some of the products developed
thus far are the following:
• Food and Gill Exchange of Toxic Substances
(FGETS) Model for estimating uptake of
neutral hydrophobic chemicals in fish.
• Approaches to assessing effects of neutral
hydrophobic chemicals on aquatic chemicals.
• Aquatic Plant Uptake Model.
Center for Exposure Assessment Modeling
• (CEAM).
Summary
In summary, I offer the following considerations:
• There is a legal and scientific basis for using
populations as the unit of ecological risk as-
sessment.
• The quotient method of ecological risk
assessment is used in the Office of Pesticides
Programs and the Office of Toxic Substanc-
es. In spite of its deficiencies, it has been
used successfully to regulate industrial chem-
icals and pesticides.
Population and ecosystem models are being employed
in our office to evaluate the consequences of exceed-
ing certain lexicological endpoints. As such, they are
currently being used as adjuncts to the quotient
method as opposed to replacing the quotient method.
Questions
Q: I had the impression that proposers of new
chemicals must bring quantities of toxicity
data for new chemicals. Is that incorrect?
A: Yes, that is incorrect.
Q: I had the impression that you go through a
full analysis independent of that.
A: Yes, we do. Because of the paucity of data
we rely on the QSAR's and SAR's, analogs,
and a lot of databases to get those estimates.
I might also add that in order to register a
pesticide (as opposed to a new chemical) a
great deal of test data is necessary.
Q: How much experience have you had with the
verification of these population and ecosys-
tem models; how confident are you in your
predictions?
A: We did an evaluation of the SWACOM
model with outdoor ponds. The models
were picking up effects as we became more
specific with the information we gave it. If
we gave it analog data, it didn't respond. As
we gave more precise data, the model pre-
dicted effects different from those observed
in the ponds. Finally when we modified the
model we got good effects — the model
predictions compared favorably to the actual
pond's effects. Assessment factors worked
10
-------
surprisingly well for the ponds. Note that
this was only one study and we are continu-
ing to compare model predictions with data
obtained from field studies.
Q: What criteria are used to draw the line in the
registration of a chemical for manufacture?
A: The first option is that testing be done by the
manufacturer, both for human health and
ecological effects. About 60% of the time
they elect to do the testing and we take it to
the point where either the concern levels
aren't being exceeded, in the case of ecologi-
cal assessment, or we're getting distinct "no
effect" levels, in the case of human health.
Rodier further explained the last answer at a later
date. He wrote: "Judging from the use of the term
'registration' there may be some confusion about my
office and OPP (Office of Pesticide Programs). OPP
registers pesticides; we [OTS] do not register indus-
trial chemicals. The basic criterion for both OTS and
OPP is that the chemical not cause unreasonable
risks to human health and the environment. In the
case of OTS, if a substance is deemed to pose a risk
to human health or the environment, testing can be
required. These tests proceed in a tiered fashion,
going from short-term acute tests to more complex
chronic tests. The outcome of the first set of tests
determines whether additional testing is necessary. In
addition to evaluating the hazard of a particular
chemical, the potential exposure of that chemical to
humans or the environment is evaluated. If the risk
assessment indicates no unreasonable risks, the
chemical is allowed to be manufactured."
References
Auer, C.M., J.V. Nabholz, and K.P. Baetcke. 1989.
Mode of action and the assessment of chemical
hazards in the presence of limited data: use of struc-
ture activity relationships (SAR) under TSCA Section
5. Environmental Health Perspectives (In Press).
Barnthouse, L.W., G.W. Suter, S.M. Bartell, J.J.
Beauchamp, R.H. Gardner, E. Under, R.V. O'Neill,
and A.E. Rosen. 1986. User's manual for ecological
risk assessment. Oak Ridge, TN: Oak Ridge National
Laboratory Publication No. 2679.
Bartell, S.M. 1987. User manual for ecosystem
uncertainty analysis: demonstration program. Wash-
ington, DC: Environmental Effects Branch, Health
and Environmental Review Division, Office of Toxic
Substances, U.S. Environmental Protection Agency
20460-0001. IAG No. DW89930690-01-0.
Bartell, S.M. 1987a. Technical reference and user
manual for ecosystem uncertainty analysis (EUA): 1)
The pascal demonstration program, 2) The standard
water column model (SWACOM), 3) The comprehen-
sive aquatic system model (CASM). Washington, DC:
Environmental Effects Branch, Health and Environ-
mental Review Division, Office of Toxic Substances,
U.S. Environmental Protection Agency. IAG No.
DW89930690-01-0.
Bartell, S.M., R.H. Gardner, and A.L. Brenkert. 1987.
Alternative models for extrapolating the effects of
phenolic compounds in aquatic systems. Washington,
DC: Environmental Effects Branch, Health and
Environmental Review Division (TS-796), Office of
Toxic Substances, U.S. Environmental Protection
Agency 20460-0001. IAG No. DW89930690-01-0.
Clements, R.G. 1983. Environmental effects of
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es, U.S. Environmental Protection Agency 20460-
0001.
11
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Clements, R.G., ed. 1988. Estimating toxicity of
industrial chemicals to aquatic organisms using
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ton, DC: Office of Toxic Substances, U.S. Environ-
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O'Neill, R.V., R.H. Gardner, L.W. Bamthouse, G.W.
Suter, S.G. Hildebrand and C.W. Gehrs. 1982. Eco-
system risk analysis: a new methodology. Environ-
mental Toxicology and Chemistry 1: 167-177.
Rodier, D.J. 1987. Ecological risk assessment in the
office of toxic substances: Problems and Progress,
1984-1987. Unpublished report dated Sept. 4, 1987.
Washington, DC: Environmental Effects Branch,
Health and Environmental Review Division (TS-796),
Office of Toxic Substances, U.S. Environmental
Protection Agency 20460-0001.
Rohlf, F.J., S. Person, and L. Ginzburg. 1986.
RAMAS 3.1 Risk analysis and management alterna-
tives system: User manual. Setauket, NY: Applied
Biomathematics. Report to Electric and Power Re-
search Institute.
USEPA. 1984. Establishing concern levels for concen-
trations of chemical substances in the aquatic environ-
ment. Washington, DC: Environmental Effects
Branch, Health and Environmental Review Division
(TS-796), Office of Toxic Substances, U.S. Environ-
mental Protection Agency 20460-
0001.
USEPA. 1985. Toxic substances control act test
guidelines final rules. Washington, DC. 40 CFR 797.
Federal Register Vol. 50, No. 188. Dated Friday,
Sept. 27, 1985.
USEPA. 1986. New chemical review process manual.
Washington, DC: Office of Toxic Substances, U.S.
Environmental Protection Agency. EPA-560/3-86-002.
USEPA. 1988. Probabilistic Dilution Model 3 (PDM-
3). Draft report dated May 31,1988. Washington, DC:
Exposure Assessment Branch, Exposure Evaluation
Division (TS-798), Office of Toxic Substances, U.S.
Environmental Protection Agency 20460-0001. Con-
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Urban, D.J., and N. Cook. 1986. Ecological risk
assessment. Washington, DC: Office of Pesticide
Programs. U.S. Environmental Protection Agency.
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Environmental Effects Branch, Health and Environ-
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Substances, U.S. Environmental Protection Agency
20460-0001.
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0001.
12
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Population Risk Assessments Based on
Toxicity Testing
Glenn W. Suter II and L. W. Barnthouse (presented
by Dr. Suter)
Defining risk assessment
Risk assessment estimates the probability of undesired
events. In practice, risk assessors attempt to quantita-
tively define the relationship between the assessment
endpoint and the available information concerning the
particular environment, the environmental chemistry
of the pollutant, and the toxicity of the pollutant.
Examples of relationships that must be inferred
include those between measured environments and the
assessed environments, between the environmental
chemistry of a chemical and its concentration in
various media (i.e., transport and fate models), and
between toxicity test endpoints and assessment
endpoints.
Risk assessments quantify uncertainty in the data and
in modeling assumptions and estimate their contribu-
tion to the uncertainty concerning the effects of the
pollution (i.e., the probability of having effects that
exceed the assessment endpoint). There are two types
of risk assessment. Prospective (or predictive) risk
assessment is useful in finding the potential effects of
new chemicals before they are released into the
environment. Retrospective risk assessment is con-
cerned with the continuing effects of events that
began in the past. Most of the concerns in the
Chesapeake Bay involve retrospective risk assessment.
For a predictive assessment, the first step is to select
endpoints of concern. These must be well-defined,
as, for example, striped bass abundance or oyster
production. Then the source terms, estimates of the
rate at which a chemical or chemical mixture is
released, are developed. The type of environment that
is of concern must be determined. Then an effects
assessment and an exposure assessment must be done
for that source term and environment. Finally, the
exposure assessment and effects assessment are
combined for an estimate of risk.
Use of whole organism toxicity data
Standard organism-level toxicity tests and endpoints
exist, but they are not designed for risk assessment.
These tests are not applicable to the very short
exposures (i.e., spills) or longer chronic exposures
that interest us. Endpoints for these tests are calculat-
ed using hypothesis testing statistics, which are
inappropriate for risk assessment. In addition, sensi-
tive and important responses may not be measured,
temporal dynamics may be neglected, and results may
be inadequately reported.
Test endpoints that reflect population properties can
be calculated. Examples include (1) weight of young
per female, (2) intrinsic rate of increase (r), and (3)
reproductive potential. In addition, tests can be
designed to tell you about the temporal dynamics of
effects. This information can then be used in popula-
tion or ecosystem models to give a good estimate of
effects.
Use of whole-organism toxicity in predictive assess-
ments. Conventional test endpoints can be used in the
quotient method to rank chemicals. In this standard
method, the quotient calculated is the test endpoint/-
estimated environmental concentration.
With use of the Analysis of Extrapolation Error
method, the probability of exceeding lexicological
endpoints can be extrapolated in several ways.
Endpoint values can be extrapolated for untested
species; the more closely related organisms yield
more similar responses. Extrapolations can be made
for different life stages of the same organism.
Temporal extrapolations under different exposure dy-
namics can give information on concentration/dur-
ation/response relationships.
13
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Populations of interest can be modeled and toxic
effects that are parameters of the models (mortality,
fecundity, growth) can be incorporated. The output of
the model is the probability of a particular level of
effect on a population attribute (e.g., probability of a
reduction in numbers of adult fish > 0.1). Four
dimensions can be addressed: concentration, duration,
severity of response, and proportion of the population
responding. The goal is to produce a response
surface incorporating severity of effect, concentration,
and duration; this response surface is the basis for
predictions.
For population models, toxic effects are measured by
parameters such as mortality, growth and fecundity.
Extrapolation analysis is used to estimate parameters
of the population model from the toxicity data.
Extrapolations from toxicity data to model parameters
are a greater source of variance in fish population
models than differences in life history or fishing
pressure.
Examples of extrapolations:
• LCSOs of Perciformes can be used to predict
LCSOs for Salmoniformes.
• An MATC (maximum allowable toxicant
concentration) can be predicted from an
LC50, but there is a lot of scatter because
the MATC is not a consistent endpoint.
• An LC50 can also be used to predict a
specific chronic effect such as an EC25 of
egg hatchability; this is a much better predic-
tion than the MATC.
An estimate of the risk of a parameter such as MATC
being exceeded in the field can be determined by the
overlap of the contaminant concentration in the field
and the MATC distribution predicted by the extrapo-
lation methods.
By looking at the fisheries data models, behavior of
the model can be statistically matched to time series.
Population models can then be used to look at the
effects that differences in life history among fishes
have on the influence of toxicity.
Ideally, we would like to do full life-cycle tests for
organisms of interest. But in lieu of this, the percent-
age reduction in abundance as a function of concen-
tration can be determined by the model. This has
confidence bounds which can be quite tight if the data
are good. But if you only have an LC50 for a
surrogate species, the confidence bounds can be up to
two orders of magnitude.
Use in retrospective assessments — assessments of
past and ongoing pollution. Population and whole-
organism responses in the field can be used to estab-
lish the nature, magnitude, and association with pol-
lutants of the assessment endpoints. Another ap-
proach is to use whole-organism responses in toxicity
tests to establish that the pollutant exposure levels in
the field could cause the effects observed in the field.
If the species, life stages, etc. in the laboratory are
different from those in the field, extrapolation models
must be used. Population and ecosystem models are
also used to match laboratory responses of individual
organisms to population-level effects in the field.
Use of suborganismal toxicity data
No consistent endpoints have been developed for
these assays, and they are not regularly used in
assessment. For use in predictive assessment, sub-
organismal responses are generally more sensitive
than conventional organismal responses, but their use
to predict effects on populations is problematic. One
reason is that their inherent importance is not clear -
nobody cares about a fish's histology or enzyme
levels per se. Also they have not generally been
shown to be correlated with responses that are impor-
tant; in many cases, the correlation has not been
investigated. A third difficulty is that there are no
14
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models to extrapolate suborganismal responses be-
tween taxa, life stages, and exposure durations.
In retrospective assessment, suborganismal toxicity
data can be of use in two ways. First, it can supple-
ment organismal and population responses by serving
as the basis for a diagnostic syndrome. It can provide
evidence of the regularity of association of a pollutant
and an effect -- evidence for which organismal and
population responses are too generic. It may also
provide a link between field observations and toxicity
tests. (If symptoms are the same then they provide
evidence of common causation.)
Suborganismal toxicity data can also be useful if the
population of interest is no longer present. In this
case the suborganismal responses of more resistant
species could provide evidence that the loss was due
to pollutant effects. This use again raises the issue of
correlation of suborganismal with organismal and
higher effects: do the suborganismal responses lead to
organismal responses? It also raises the issue of ex-
trapolation again: is it credible that a more sensitive
species would become extinct at a pollutant exposure
level that caused the observed suborganismal respons-
es in the presumed resistant species? The answers to
these questions must come from toxicity testing. One
cannot expect to observe a population as it is under-
going a pollution-induced crash and measure subor-
ganismal responses.
Use of sediment toxicity data
In theory, organism-level data in sediment would be
treated like organism-level data in water, and subor-
ganismal data would also be treated the same in all
media. In practice, only the quotient method and
similar nonpredictive approaches have been used with
sediment toxicity data. This is partly because the
problem of exposure assessment has not been re-
solved. Likewise, appropriate assessment endpoints
have not been established. For instance, are we con-
cerned about inherently valuable benthic species? Are
we concerned about food species for fish? This
matter of defining endpoints is a regulatory and
political concern as much as a scientific one. We
must remember that what we are working for in risk
assessment is a regulatory tool.
Questions
Q: Have you used additivity models to predict
interactions between multiple components?
A: Yes, we use concentration additivity models
because of Lloyd's evidence that the most
common mode of chemical interaction is
additivity and that synergisms are extremely
rare or nonexistent.
Q: How do you calculate probability when you
extrapolate more than once; for example,
when you go from LCSOs of one species to
some other measure of toxicity?
A: There are two ways to do that. First, one
can directly regress the chronic response, the
MATC for the species you are interested in,
against the LC50 for the test species and
skip all the intermediate stages. Use the
initial X and the final Y. Second, one can
perform the extrapolation in multiple steps
and carry the variance from one step to the
next.
Q: But how do you determine the error you are
losing at each step?
A: Well, you are not losing anything the first
way because you are directly regressing the
endpoint response against the measured
response. For the second method, you can
add the variances of each of the regression
steps.
15
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Q: Are you saying that suborganism level pro-
grams are not useful because you cannot
connect with populations or are you just
saying that these experiments haven't been
done?
A: Identification of endpoints is more a regula-
tory and political process than a scientific
one. The political process will decide this.
A: The latter. I hope they'll be done, in order
to allow us to make these extrapolations.
Q: But don't you need that kind of information
to really understand the population?
A: Yes, but you don't know whether the animal
is actually going downhill, or if it is just
adapting and it will continue to live and
reproduce. This will be valuable if it can be
shown to have effects on the organisms as a
whole.
Q: Are parameters other than reproduction,
growth, and mortality such as those that
affect recruitment being taken into account?
A: Recruitment is a function of reproduction,
growth, and mortality. The example you
mentioned, predation on the larvae of inter-
tidal invertebrates as they pass through kelp
forests, is simply a specific ecosystem-level
mechanism influencing one of the population
parameters. To explicitly incorporate preda-
tion, competition, etc., you would need an
ecosystem model. These processes are
implicit in the population model's backg-
round survivorship.
Comment from audience: The bottom line is really
the number of organisms that enter the population.
Q: What is an acceptable endpoint for risk
assessment in terms of population impact?
Is it a 5% reduction in community structure?
Until that is determined, modeling processes
have very little meaning.
16
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Plenary Session B:
Sediment Toxicity Assays
Convener: Ray Alden
Considerations for Sediment Toxicity
Tests
John Scott
Introduction
The questions and issues pertaining to sediment
toxicity assays that this workshop has been asked to
address are:
8) How do sediment toxicity test results apply
to risk characterization?
Toxicity tests are quantitative measures of contami-
nant impacts on survival, behavior, genetic processes,
physiology, reproduction, and other biological pro-
cesses of higher order. They are used to assess
media-specific toxic effects and can also provide an
integrated response to complex mixtures.
1) What routes of exposure of contaminants
should be assessed?
2) What toxicity test methods are currently
available?
3) How can these methods be applied to the
Chesapeake Bay system and what is the
feasibility of these methods for large-scale
monitoring?
4) What species should be selected for these
assays and what should the selection criteria
be?
5) What biological endpoints should be selected
that would be appropriate?
6) Can one extrapolate among responses, i.e.,
from suborganismal to acute and chronic
responses to resources at risk?
7) Sample collection and experimental design
issues will be addressed at the workgroup
sessions.
The responses most commonly and historically used
have included lethality and generation of LCSOs.
More recently methods have been developed to
examine sub- and supra-organismal responses.
Sediments are important because they can act as long-
term reservoirs of dissolved and particulate-associated
contaminants, and benthic organisms are a major
vector in food chain transfer and biomagnification.
There are many factors controlling contaminant
availability and, hence, toxicity in natural systems.
There is biological mediation of contaminant flux via
biodeposition, bioturbation, and bioaccumulation.
More information is needed to understand sediment
contaminant availability and how it is influenced by
factors such as organic carbon content, grain size,
water content, and oxidation state.
Sediment toxicity tests have had many applications.
The primary impetus for sediment testing has been
through the regulatory process, specifically for
dredge-material permitting, where a recommended
suite of tests were put forth by the EPA and the
Corps of Engineers in the 1977 "Green Book."
These tests commonly used a burrowing clam, a
17
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burrowing polychaete, and epibenthic shrimp or fish
for 10-day exposures, and then quantified survival and
bioaccumulation. Sediment toxicity tests have been
used widely on the West Coast (e.g., Puget Sound,
San Francisco Bay) to monitor spatial and temporal
trends of contaminant effects in sediments. A subset
of this type of monitoring deals specifically with
remedial action studies at Superfund sites, identifying
hot spots and looking at the condition of sediments as
a result of source inputs. Finally, sediment toxicity
tests are used in the research arena to examine
modes of toxicity and contaminant-specific exposures
that result in tissue-specific pathologies or physiologi-
cal responses. They are being used to define extrapo-
lation potential across the biological hierarchy, from
sub- to whole- to supra-organismal responses. These
methods are also being used to evaluate contaminant
bioavailability through tests with spiked sediments
and field sediments.
An example of how sediment data are used to define
spatial gradients is drawn in Figure 1. These are test
results for 10-day exposures to New Bedford Harbor,
MA sediments using the amphipod Ampelisca abdita.
Figure 1.
AMPELISCA ABDITA
NEW BEDFORD HARBOR GRADIENT TEST
CUMULATIVE DAILY MORTALITY
These stations represent a gradient of PCB and heavy
metal contamination increasing up the harbor from the
lower-numbered to the higher-numbered stations (Fig-
ure 2). The mortality pattern clearly mimics the
degree of contamination.
Flfur«2.
AEROVOX
-245000
ACUSHNET
-240000
-235000
NEW
-230000 BEDFORD
-225000
COGGESHALL ST.
RTE 195
FAIRHAVEN
HURRICANE
.,_ BARRIER
CORNELL
UBIUEr,,
10
(DAYS)
Sediment toxicology is a relatively new field. Prior
to 1975, sediment toxicity was generally inferred from
field studies. In 1977, the EPA/COE developed an
implementation manual that provided general guide-
lines for evaluating proposed dredged materials.
Because the limitations of existing methods were
recognized at that time, this manual was the impetus
for sediment toxicity research and the development of
standard methods within EPA as well as the COE's
Dredged Material Research Program (DMRP). The
early 1980s saw broadscale applications of sediment
tests, particularly Swartz and colleagues' development
of the Rhepoxynius abronius amphipod test method
(Swartz et al., 1985). Further research has led to an
18
-------
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expansion of test species and endpoints, and the
examination of modes of toxicity and contaminant
availability. In the late 1980s we see widespread
application of new methods, a growing understanding
of modes of toxicity and the factors influencing
availability in complex mixtures (i.e., synergistic and
antagonistic effects), and the development of sediment
quality criteria.
Exposure assessment and test applications
Several different media have been used in sediment
toxicity tests. Ibcse include whole sediments, sus-
pended sediments, pore waters, elutriates, and ex-
tracts. Methods of obtaining pore waters include
centrifuging, squeezing, and applying pressure.
Extracts may be prepared chemically using organic
solvents. In most cases, whole sediment exposures
19
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are the preferred method because pore water, extract,
and elutriate methods suffer from interpretation
problems. Suspended sediments are typically not an
exposure route of concern.
Hazard assessment
Distinctions commonly used (often interchangeably)
in hazard assessments are: acute vs. chronic (with the
implication of time and an effects component); short-
vs. long-term; lethal vs. sublethal; and screening vs.
interpretative assays (with the latter implying the
occurrence of a response to be interpreted). I suggest
that the workgroup consider these definitions, but in
any case, see Chapman (1989).
The range of biological responses that have been
measured in tests with sediments spans the biological
hierarchy. They include biochemical, cytogenetic,
pathological, immunological,and physiological re-
sponses, as well as survival, growth, behavior, repro-
duction, development, metamorphosis, population
growth, recruitment, and community structure. A
series of screening level tests have been developed
with bacteria (of which Microtox is an example) that
examine genetic damage and bioluminescerice. The
types of biological responses that could be evaluated
are summarized in Figure 3 (after Sheehan 1984). As
one progresses up the biological hierarchy, the time
scale of response increases from hours to years. In
this progression, however, predictive value and
ecological relevance also increase. Ideally tests
should have reasonably short time periods but also
have predictive value.
The EPA/COE Field Verification Program allowed for
examination of a series of endpoints along this hierar-
chy using several different test species. In the most
sensitive species tested, Ampelisca abdita, effects
were seen on pathology, growth, survival, behavior,
reproduction, and intrinsic rate of population growth.
The latter responses were the most sensitive but also
took approximately two months to develop.
Criteria for test organism selection
Test organisms should be selected on the basis of the
following criteria:
• Ready availability either by culture or collec-
tion
• Wide geographic distribution
• Taxonomic relationship to site inhabitants
• Demonstrated sensitivity
• Ease of laboratory maintenance
• Ease of test method
• Ecological and/or economic relevance (most
important)
• Compatibility with exposure/response matrix
• Insensitivity to geophysical factors
• Cost effectiveness
Available methods that may be appropriate to the
Chesapeake Bay include the 10-day amphipod test,
which has been used primarily in West Coast applica-
tions. One could also try several ranges of responses
and tests, such as those proposed by Long and Chap-
man in the sediment quality triad using a suite of tests
where various exposures and endpoints are evaluated.
Application to risk characterization
In order to apply sediment toxicity tests to risk
characterization, we must determine the extrapolation
potential of such tests. Specifically, (1) how do we
extrapolate from a laboratory test to a field situation
and (2) how do we extrapolate across the biological
hierarchy from lower to higher levels of organization?
20
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Both questions pose serious challenges. As far as
data needs, extrapolation from the laboratory to the
field requires good contaminant exposure information,
which is based on local hydrography and circulation,
and contaminant distribution. The biological resourc-
es at risk need to be identified; measured laboratory
effects must be linked to these resources.
One quick example of the application of laboratory
responses to field predictions using A. abdita is field
verification at the Black Rock Harbor disposal site in
Central Long Island Sound.
Flflur* 4.
AMPELISCA ABDITA
100
ACUTE MORTALITY
CHRONIC MORTALITY
GROWTH
A POPULATION GROWTH
Figure 4 shows the ratio of predicted field exposures
of contaminated suspended sediments to the effect
concentrations for various responses. Exposure is
defined and hazard levels are identified; they are
combined using the quotient method (exposure/res-
ponse) to derive risk predictions. This figure shows
the potential for risk to each of the endpoints of
concern. While these estimates have a great deal of
associated uncertainty, the point here is that the
approach and methods are available. To summarize,
the toxicity test has to consider the type of organism,
the response, and the exposure. The toxicity tests
must link to field exposures and ecological resources
in a predictive way.
Issues and research
Research must deal with sediment holding times,
sediment manipulation, assumptions about chemical
equilibrium, low-salinity tests, site specificity, and
chronic test methods. We should also continue to
investigate how to interpret and extrapolate suborgan-
ismal responses.
Questions
Q: Do we agree on cross-species sensitivity?
A: Two amphipods (R. abronius and A. abdita)
have similar sensitivity; not many species
have been tested for relative sensitivity.
Q: What about Microtox uses?
A: The problem is, what are you testing? If
you're not using whole sediments, the inter-
pretive perspective is lost.
Q: It seems that population growth is a more
sensitive response than others.
A: This is true, but, as noted, these responses
require a longer time frame. The combina-
tion of a series of responses together gives
more complete information than any single
response.
Q: How long until we have sediment quality
criteria?
21
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A: That's a good question, and the next speaker
will address it.
References
Chapman, P.M. 1988. Marine sediment toxicity tests.
pp. 391-402. In J. J. Lichtenberg, F. A. Winter, C. I.
Weber and L. Fredkin [eds]. Chemical and biological
characterization of sludges, sediments, dredge spoils
and drilling muds. ASTM STP 976, Philadelphia,
PA.
Chapman, P. M. 1989. A bioassay by any other
name might not smell the same. Env. Toxicol. Chem.
8:551.
Environmental Protection Agency/Corps of Engineers.
1977. Ecological evaluation of proposed discharge of
dredge material into ocean waters. U. S. Army
Engineers' Waterways Experiment Station, Vicksburg,
MS.
Lee, H. L., and R. C. Swartz. 1980. Biological
processes affecting the distribution of pollutants in
marine sediments. Part II. Biodeposition and biotur-
bation. pp. 555-606. In R. A. Baker, [ed.] Contami-
nants and sediments, Vol. 2. Ann Arbor Science
Publishers, Ann Arbor, MI.
Long, E. R., and P.M. Chapman. 1985. The sedi
ment quality triad: measures of sediment contamina-
tion, toxicity, and informal community composition in
Puget Sound. Mar. Poll. Bull. 16: 405-415.
Rhoads, D. C., and L. F. Boyer. 1982. The effects
of marine benthos on physical properties of sedi-
ments, pp. 3-52. In P. L. McCall and M. J. Tevesz
[eds.] Animal-sediment relations. Plenum Press
Geobiology Series, New York.
Scott, K. J., and M. S. Redmond. 1989. The effects
of a contaminated dredged material on laboratory
populations of the tubiculous amphipod Ampelisca
abdita. pp. 289-303. In U. M. CowgiU and L. R.
Williams [eds.] Aquatic Toxicology and Hazard As-
sessment: 12th Vol. ASTM. Philadelphia, PA.
Sheehan, P. J. 1984. Effects on individuals and
populations, pp. 23-50. In P. J. Sheehan, D. R.
Miller, G. C. Butler, and P. Bourdeau [eds.] Effects
of pollutants at the ecosystem level. John Wiley and
Sons, Ltd., New York.
Swartz, R. C. 1987. Toxicological methods for
determining the effects of contaminated sediment on
marine organisms, pp. 183-198. In K. L. Dickson,
A. W. Maki, and W. A. Brungs [eds.] Fate and
effects of sediment bound chemical in aquatic system.
Pergamon Press, New York.
Swartz, R.C., W.A. DeBen, J.K. Jones, J. O. Lamber-
son, and F. A. Cole 1985. Phoxocephalid amphipod
bioassay for marine sediment toxicity. pp. 284-307,
In R. D. Cardwell, R. Purdy, and R. C. Bahner
[eds.]. Aquatic toxicology and hazard assessment,
seventh symposium, ASTM STP 854. ASTM,
Philadelphia, PA.
Swartz, R. C., and H. F. Lee. 1980. Biological
processes affecting the distribution of pollutants in
marine sediments. Part I. Accumulation, trophic
transfer, biodegradation and migration, pp. 533-553.
In R. A. Baker, [ed.] Contaminants and sediments,
Vol. 2. Ann Arbor Science Publishers, Ann Arbor,
MI.
22
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Contaminated Sediments and Sediment
Criteria
Christopher S. Zarba
Introduction
Toxic contaminants in bottom sediments of the United
States' lakes, rivers, and coastal waters create the
potential for continued environmental degradation
even where water column pollutant levels comply
with established water quality criteria. The absence
of defensible numerical sediment quality criteria
makes it difficult to accurately assess the extent of the
contaminated sediment problem. However, existing
data demonstrate that there are many locations where
existing and projected sediment contaminant concen-
trations are causing significant adverse effects to
aquatic life and human health. The application of
sediment criteria will make it possible for the States
and the Environmental Protection Agency (EPA) to
more effectively implement regulatory, enforcement,
and cleanup actions where necessary.
Authority
Under the Clean Water Act (CWA) EPA is responsi-
ble for protecting the chemical, physical, and biologi-
cal integrity of our nation's waters. In keeping with
this responsibility, EPA published ambient water
quality criteria for the 65 priority pollutants and
pollutant categories listed as toxic in the CWA.
While ambient water quality criteria are playing an
important role in assuring a healthy aquatic environ-
ment, they alone have not been sufficient to ensure
appropriate levels of environmental protection.
EPA has authority to pursue the development of
sediment criteria in streams, lakes and other waters of
the United States under sections 104 and 304(a)(l)
and (2) of the CWA as follows:
(1) Section 104 authorizes the Administrator to
establish national programs for the prevention, reduc-
tion and elimination of pollution by conducting and
promoting "the coordination and acceleration of
research, investigation, experiments, training, demon-
strations, surveys, and studies relating to the causes,
effects, and extent, prevention, reduction, and elimina-
tion of pollution and by publishing relevant informa-
tion. Section 104(n)(l) specifically provides for the
study of the effects of pollution, including sedimenta-
tion, in estuaries on aquatic life."
(2) Section 304(a)(l) directs the Administrator to
develop and publish criteria for water quality accu-
rately reflecting the latest scientific knowledge "on the
kind and extent of all identifiable effects on health
and welfare including, but not limited to, plankton,
fish, shellfish, wildlife, plant life, shorelines, beaches,
aesthetics, and recreation which may be expected
from the presence of pollutants in any body of water,
including groundwater ... on the concentration and
dispersal of pollutants, or their by-products ... on the
effects of pollutants on biological community diversi-
ty, productivity and stability, including information on
the factors affecting .. . rates of organic and inorgan-
ic sedimentation for varying types of receiving
waters."
(3) Section 304(a)(2) directs the Administrator to
develop and publish information on, among other
things, "the factors necessary for the protection and
propagation of shellfish, fish, and wildlife for classes
and categories of receiving waters. . ."
(4) To the extent that sediment criteria could be
developed which addressed the concerns of the
section 404(b)(l) Guidelines (for discharges of
dredged or fill material under the Clean Water Act) or
ocean dumping criteria (under the Marine Protection,
Research, and Sanctuaries Act), they may also be
incorporated into those regulations."
23
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Approach to Criteria Development
A workshop was convened in which a wide variety of
experts were given the responsibility of identifying a
preferred method for generating sediment criteria.
When potential approaches were being considered for
their utility in providing an effective regulatory tool,
it was understood that each approach had strengths
and weaknesses and that any one approach would not
be best for all situations. The participants of this
workshop were provided with documents that:
• identified EPA's legal authority to develop
sediment criteria and to regulate through
their use, and
• identified a variety of methods that could be
used in the development of sediment criteria,
and
• identified the findings of a study that looked
at contaminated sediments on a national
basis.
A preferred approach was selected that many believed
would provide EPA with the most effective regulatory
tool. The Equilibrium Partitioning Approach (EP)
was selected as the approach to be pursued to meet
this goal. Over the past several years activities have
been focused on evaluating and developing the EP for
generating sediment criteria for regulatory purposes.
Description of Method
The EP focuses on predicting the chemical interaction
between sediments and contaminants. An understand-
ing of the principal factors that influence the sedi-
ment/contaminant interactions allows predictions to be
made (based on sediment chemistry analysis) about
the concentration of contaminant to which benthic and
other organisms may be exposed. Data have demon-
strated that the concentration of contaminant in the
interstitial water (water between the particles of
sediment) correlates very closely with toxicity and
that the concentration of contaminant on the sediment
does not. Chronic water quality criteria or possibly
other lexicological endpoints, when compared with
the concentration of contaminant in the interstitial
water, could then be a reliable predictor of potential
biological effects. The EP for generating sediment
criteria focuses on predicting the concentration of
contaminant in the interstitial water and compares that
concentration to quality criteria. If the predicted
sediment interstitial water concentration for a given
contaminant exceeds the chronic water quality criteria
for that contaminant then the sediment would be
expected to be causing adverse effects.
The principal factors that influence sediment/contam-
inant interactions vary with the types of contaminants
involved. Non-ionic, ionic, and metal contaminants
interact with sediments in different ways. For non-
ionic organic contaminants, predictions of sediment/-
contaminant interactions are dependent on organic
carbon as the principal factor that influences sediment
contaminant binding. Sulfur compounds and organic
carbon are some of the key factors that are being
investigated that may influence binding between metal
contaminants and sediments. Polar organic contami-
nants are currently being investigated to determine
their chemical interaction with the sediments.
Specific Applications
Specific applications of sediment criteria are under
development. The primary use of EP-based sediment
criteria will be to assess risks associated with contam-
inants. The various offices and programs concerned
with contaminated sediment have different regulatory
mandates and thus have different needs and areas for
potential application of sediment criteria. Because
each regulatory need is different, EP-based sediment
quality criteria which are designed specifically to
meet the needs of one office or program may have to
24
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be implemented in different ways to meet the needs
of another office or program.
A likely mode of application of EP-based numerical
sediment quality criteria would be a tiered approach.
With such an application, the sediments would be
considered to cause unacceptable impacts when
contaminants in sediments exceed the sediment
quality criteria. Further testing may or may not be
required depending on site-specific conditions.
Contaminants in a sediment at concentrations less
than the sediment criteria would not be of concern.
However, in some cases the sediment could not be
considered safe because they may contain other
contaminants above safe levels for which no sediment
criteria exist. In addition, the synergistic, antagonis-
tic, or additive effects of several contaminants in the
sediments may be of concern. Additional testing in
other tiers of the evaluation approach, such as bio-
assays, could be required to determine if the sediment
is safe. It is likely that such testing would incorpo-
rate site-specific considerations. At the present time
standard bioassays for assessing sediment contamina-
tion on a national basis are under development.
Current Use
The specific regulatory uses of EP-based sediment
quality criteria have not been established. A review
of the method for generating sediment criteria for
non-ionic contaminants by the Science Advisory
Board is ongoing. It is intended that this review be
completed prior to the establishment of any formal
framework for the application of sediment criteria.
(The review of the method was held on February 2,
1989. The findings of this review are expected in
September 1989.) The range of potential applications
is quite large since the need for the evaluation of
potentially contaminated sediments arises in many
contexts.
Interim sediment criteria values were developed using
the EP approach for a variety of organic compounds
and were used to assist in the decision-making pro-
cess in a pilot study involving six sites. These sites
were Superfund sites that were involved with site
characterization and evaluation activities. The interim
criteria were used to:
«• identify the extent of contamination
• assess the risks associated with the sediment
contamination
• identify the environmental benefit associated
with a variety of remedial options.
Potential Use
The EP method is likely to be useful in many of the
activities being pursued by EPA. EP-based sediment
quality criteria could play a significant role in the
identification, monitoring, and cleanup of contaminat-
ed sediment sites on a national basis and in ensuring
that uncontaminated sites will remain clean. In some
cases sediment criteria alone would be sufficient to
identify and to establish cleanup levels for contami-
nated sediments. In other cases the sediment criteria
would be supplemented with biological sampling and
testing or other types of analysis before a decision
could be made. Sediment criteria can provide a basis
for determining whether contaminants are accumulat-
ing in sediments to the extent that an unacceptable
contaminant level is being approached or has been
exceeded. By monitoring contaminants in the vicinity
of a discharge, contaminant levels can be compared to
sediment criteria to assess the likelihood of impact.
EP-based sediment criteria will be particularly valu-
able for monitoring sites where sediment contaminant
concentrations are gradually approaching a criterion
over time. Comparison of field measurement to
sediment criteria will be a reliable method for provid-
ing early warning of a potential problem. Such an
early warning would provide an opportunity to take
corrective action before adverse impacts occur.
25
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In many ways sediment criteria developed using EP
are similar to existing water quality criteria. Howev-
er, in their application they are likely to vary signifi-
cantly. Contaminants at levels of concern in the
water column need only be controlled at the source to
eliminate unacceptable adverse impacts in most cases.
Contaminated sediments have often been in place for
quite some time and controlling the source of that
pollution (if the source still exists) will not be suffi-
cient to alleviate the problem. The problem is com-
pounded by the difficulty and expense involved in
safe removal, treatment, or disposal of contaminated
sediments. For this reason sediment criteria are not
anticipated to be used as mandatory cleanup levels,
but as means for predicting or identifying the degree
and spatial extent of contaminated areas so that
regulatory decisions can be made.
Contaminated Sediment Committees
The development of sediment criteria using EP is only
one of many Agency activities that address contami-
nated sediment problems. To ensure consistency and
effectiveness in the development and implementation
of these methods and procedures two committees have
been established. These are the Contaminated Sedi-
ment Steering and Technical Committees. The
principal roles and responsibilities of these commit-
tees are as follows:
explain/resolve inconsistencies in present
sediment program activities
coordinate with other federal agencies
determine the role of economics in contami-
nated sediment strategy
identify changes needed in current statutes
Contaminated Sediment Technical Com-
mittee
coordinate technical activities
prepare guidance documents
prepare options documents
perform policy analysis
provide agency-wide technical support.
Questions and Answers
Q: What are the characteristics that affect re-
lease of contaminants?
Contaminated Sediment Steering Commit-
tee
develop long-term management strategy for
contaminated sediments
facilitate the commitment of resources
establish policy/interim guidelines for man-
agers
develop long-term research programs
A: They vary with different contaminants and
different sediments. Many factors affect
release; several factors dominate the amount
of release. For non-ionic organic contami-
nants, organic carbon is the main factor. For
metal contaminants, sulfur compounds ap-
pear to have the most influence.
Q: How do you define uncertainty?
A: We are in the process of doing that at this
time. We will develop recommendations on
this point by the end of this fiscal year.
26
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Q: Can different types of carbon affect the
availability of some contaminants?
A: Studies that we have done indicate that for
the most part carbon is carbon with the ex-
ception of large particles like coal or other
material. We are conducting additional
investigations in this area.
Q: Have you planned to include a standard
sediment test?
A: No, not for the purposes of developing
criteria. There may be potential for using
this technique when assessing variability in
sediment methods in the future.
Q: Are biological tests run along with the sedi-
ment chemical analysis?
A: Yes, this is routine.
27
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Plenary Session C:
Methodologies for Whole Organism Toxicity Testing
Convener: Lenwood Hall
Laboratory Testing of Ambient Receiving
Waters
Steven C. Schimmel
Receiving water collection and holding techniques
Evaluating toxicity of ambient receiving water in the
laboratory requires different techniques for collecting
water in different settings. The most common tech-
nique is grab sampling. Another is composite collec-
tion, which uses a mechanical compositor to collect a
sample over time. With both methods, the collector
must assure the representativeness of the sample.
Additional questions concern the different methods.
Should I collect a grab sample at the surface or take
multiple-depth collections? Will a collection of
composite samples lose toxicity under the confined
"artificial" conditions?
From our experience, ambient toxicity has been
generally near-field in nature, and is generally not as
widespread as we may expect. This is a result, in
part, of toxicity decaying over time. If an effluent is
the cause of toxicity, and it decays rapidly, then that
toxic effluent component in the water will also decay
rapidly. We therefore need to convey the sample
from the collection point to the testing point as
quickly as possible.
Sampling capabilities and options vary depending on
the environment ~ freshwater vs. saltwater, estuaries
vs. streams. Small riverine situations are amenable to
composite collection. They have accessible river
banks for placing a compositor, and provide precisely
identifiable locations. Flow conditions can also be
helpful, as the collector may take advantage of uni-
directional flow in deriving estimated effluent concen
trations. Sampling on estuaries, large lakes, and large
rivers is more difficult. Few stationary platforms are
readily available, and for estuaries and lakes, hydro-
logic patterns are often uncertain or unknown.
Exposure concentrations of chemicals or effluents in
these systems are often vaguely defined, requiring dye
studies.
These problems help us identify research needs for
collecting ambient composite samples. We need
compositors that maintain sample integrity, and the
devices must be practical. They should be deployable
in situ, function reliably in rough open water, and
should be relatively inexpensive.
Sample collection, shipment, and holding condi-
tions
Large volume samples may be necessary for toxicity
testing.
Regardless of future research needs, all ambient
samples should be held on ice for shipment to the
testing facility. They should be placed in inert,
unbreakable containers and kept in the dark. Tests
should be conducted within 36 hours of collection.
Remember that a receiving water may or may not be
toxic, but whatever toxicity is there, you want to be
able to detect.
Criteria for candidate test methods
One purpose of this workshop is to define a series of
toxicity test methods for ambient waters of the Chesa-
peake Bay watershed. There are several criteria that
should be included in that selection:
29
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Sensitivity: The endpoint must be sensitive to low-
level toxicity. It should include the most sensitive
life-stages, and sublethal endpoints. The widely-
used 96-hr acute lethality tests will not be sensi-
tive enough.
Efficiency: Tests should be quick, require a low
volume of water, and be cost-effective.
Relevance: The species chosen should be relevant
to Chesapeake Bay, if at all possible. Ideally, a
suite of phylogenetic groups could be selected that
represent the biota in the Bay. Species that
occupy important ecological niches would be
particularly desirable.
Recommended species
Freshwater:
Green alga - Selenastrum capricornutum
Duckweed - Lemna minor (number of fronds and
chlorophyll content)
Daphnid - Ceriodaphnia dubia (growth, survival,
and reproduction including "r " and time to onset
of reproduction)
Fathead minnow - Pimephalespromelas (growth,
survival)
Saltwater:
Diatom - Skeletonema costatum
Red macroalga - Champia parvula (sexual repro-
duction)
Sea urchin - Arbaciapunctulata (2-hr test, fertil-
ization)
Bivalve larvae test (48-hr test) - Crassostrea,
Mytilus and Mercenaria
Mysid - Mysidopsis bahia (growth, survival,
fecundity) - 7-day labor-intensive test
Inland silverside - Menidia beryllina - the only
species that can tolerate the salinity range of the
entire Chesapeake Bay (growth and survival)
Sheepshead minnow - Cyprinodon varlegatus
(growth and survival)
Precision testing has been conducted for most of these
species to test reproducibility. Where there are data,
variability was relatively small (30-50%). These
methods have been used to test ambient waters and
effluents in mobile laboratories around the country.
Regulatory history
In addition, all the methods listed above have been
used for regulatory purposes, that is, either in the
NPDES permitting system, for effluent toxicity
limitations, or in pesticide registration. They will
likely be used in some Superfund applications.
Acute lethality tests have the longest track record.
Most of the acute lethality methods have been sub-
jected to inter- and intra-laboratory testing programs
for precision testing. Most methods have received
legal challenges, and these challenges are ongoing.
Ambient in situ salt water methodologies
Evaluating the toxicity of ambient waters directly in
situ provides obvious advantages over laboratory eval-
uations. The blue mussel, Mytilus edulis, has been
used for that purpose for over ten years. The blue
mussel has a range that borders on the Chesapeake
Bay. About 100 organisms are placed in each basket
and suspended in the water column, where they are
kept in situ for at least one month. Measurements of
growth and survival are taken. In addition, laboratory
toxicity tests such as "scope for growth" can be done
immediately following field exposure. Scope-for-
growth measurements obtain an energy budget for the
animal to determine if energy is available for growth
and reproduction, over and above that needed for
basic metabolism.
In an in situ study, you have to make sure that you
match physical conditions such as salinity, tempera-
ture, and food availability in order to make inferences
about toxicity among stations.
-------
Questions
Q:
A:
Does the insensitivity of Cyprinodon
extend across all chemical classes?
No. You can't make any hard and fast
assumptions on sensitivity, which is very
specific to the experiment and the chemi-
cal that the species is being exposed to.
Relatively speaking, Cyprinodon and
Menidia are at the low end of the sensi-
tivity scale. For this reason and because
of the high labor intensity of this experi-
ment, this test may not be worth the
effort, depending on specific program
needs.
Would you object to the use ofNeomysis
amerlcana as opposed to Mysidopsis
bahia, since Neomysis is more relevant to
the Bay?
No, I don't have any objections. Howev-
er, we were not as successful at culturing
that species as we were with Mysidopsis.
There is still much basic work to be done
to better understand optimum culture
techniques. The listing I provided should
simply lay the groundwork, and I would
expect that a list of recommendations for
research initiatives on additional species
will come out of this workshop.
Is there a database on the relative sensi-
tivity of the organisms that you present-
ed?
Yes, there is one for the saltwater species
I discussed, with the exception of the
bivalve larvae.
Q: Have you compared the reproductive
indicator, the 7-day Mysidopsis test, to
the results you get in the 28-day test?
A: Those comparisons have been conducted
for at least 10 different organic and inor-
ganic chemicals independently; a large
majority show close correspondence (i.e.,.
good predictions of life cycle effects).
Q: Have you ever seen effluents or receiving
waters increase in toxicity over time?
A: No, I have never seen it.
Q: Are you suggesting a battery of tests as
opposed to a tiered approach? And what
are the advantages and disadvantages of
each?
A: We are evaluating toxicities in ambient
receiving waters, which do not integrate
toxicity. They may appear to be spurious
or concurrent with episodic events such
as storms and the like. Resuspension of
sediment material may cause some toxic-
ity. You're talking about short-term
exposures and endpoints that relate to life
cycles. Also, using a suite of organisms
that occur naturally is the best way to go.
A tiered approach, as is used for pesti-
cide registration, is good for a single,
conservative chemical. I don't think it is
good for ambient water that is not con-
servative. The group can decide whether
that is the best approach or not.
Q: How many chemicals would we need to
test to get a good idea of the effects?
A: I would rather look to the classes of
chemicals, i.e. organic chemicals with
31
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high vs. low octanohwater partition coef-
ficients, metals, surfactants. Also, I'd
use those chemicals shown to cause
environmental problems.
Q: Wouldn't it also be useful to use a
benchmark toxicity test - for example,
tests that we have a large database on, to
run side by side with the selected meth-
odologies for a few times?
A: Yes, one that's been used in the regulato-
ry process successfully.
This group should be aware of EMAP ~ Environmen-
tal Monitoring and Assessment Program. This
monitoring program will be carried out next summer
in a pilot study at a series of stations from Cape Cod
to Cape Hatteras including several stations in the
Chesapeake Bay. I would propose that there be good
communication between this workshop and the
EPA/NOAA EMAP program. It only makes sense to
do so.
References
American Society for Testing and Materials. 1980.
Standard Practice for Conducting Static Acute Toxici-
ty Tests with Larvae of Four Species of Bivalve
Molluscs. In: 1988 Annual Book of ASTM Standards
Section 11. Water and Environmental Technology.
284-295.
Hughes, Melissa M., M.A. Heber, G.E. Morrison,
S.C. Schimmel, and W.J. Berry. (1989) An Evalua-
tion of a Short-Term Effluent Toxicity Test using
Sheepshead Minnow (Cyprinodon variegatus) larvae.
Environ. Poll. (In Press)
Mount, Donald I. and T. J. Norberg. 1984. A Seven-
Day Life-Cycle Cladoceran Test. Environ. Toxicol.
Chem. 3: 425-434.
Nacci, Diane, Eugene Jackim, and Raymond Walsh.
1986. Comparative Evaluation of Three Rapid Marine
Toxicity Tests: Sea Urchin Early Embryo Growth
Test, Sea Urchin Sperm Cell Toxicity Test and
Microtox. Environ. Toxicol. Chem. 5: 521-525.
Nelson, William G. (Manuscript Submitted to ASTM)
The Use of the Blue Mussel, Mytilus edulis, in Water
Quality Based Toxicity Testing and in situ Marine
Biological Monitoring. ERL-Narragansett Contribu-
tion # 1022. 23pp.
Norberg, Teresa J. and D.I. Mount 1985. A New
Fathead Minnow (Pimephales promelas) Subchronic
Toxicity Test. Environ. Toxicol. Chem. 4:(5) 711-
718.
Schimmel, Steven C., George E. Morrison and
Margarete A. Heber. 1989. Marine Complex Effluent
Toxicity Testing Program: Test sensitivity, Repeat-
ability and Relevance to Receiving Water Toxicity.
Environ. Toxicol. Chem. 8:(8). (In Press)
Steele, Richard L. and Glen B. Thursby. 1986.
Laboratory Culture of Gametophytic Stages of the
Marine Macroalgae Champia parvula (Rhodophyta)
and Laminaria saccharina (Phaeophyta). Environ.
Toxicol. Chem. 7: 997-1002.
Thursby, Glen B. and Richard L. Steele. 1986.
Comparison of Short- and Long-Term Sexual Repro-
duction Tests with the Marine Red Alga, Champia
parvula. Environ. Toxicol. Chem. 5: 1013-1018.
Taraldsen, James and T.J. Norberg-King. (manu-
script). A New Method for Determining Effluent
Toxicity using Duckweed. U.S. EPA Laboratory, Du-
luth, MN.
U.S. Environmental Protection Agency 1988. Short-
Term Methods for Estimating the Chronic Toxicity of
Effluents and Receiving Waters to Marine and Estuar-
ine Organisms. EDS. C.I. Weber, W.B. Horning II,
32
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DJ. Klemm, T.W. Neiheisel, P.A. Lewis, E.L.
Robinson, J. Menkedick, and F. Kessler. EPA/600/4-
87/028. 417p.
U.S. Environmental Protection Agency. 1989. Short-
Term Methods for Estimating the Chronic Toxicity of
Effluents and Receiving Waters to Freshwater Organ-
isms. 2nd Ed. C.I. Weber, W.H. Peltier, T.J. Norberg-
King, W.D. Horning, II, F.A. Kessler, J.R.Menkedick,
T.W. Neiheisel, P.A, Lewis, DJ. Klemm, Q.H.
Pickering, E.L. Robinson, J.M. Lazorchak, L.J.
Wymer and R.W. Freyberg. EPA/600/4-89/001. 249p.
Widdows, J., O.K. Phelps, and W.B. GaUoway. 1981.
Measurement of the Physiological Condition of
Mussels Transplanted Along a Pollution Gradient in
Narragansett Bay. Marine Environ. Res. 4: 181-194.
Woelke, C.E. 1972. Development of a Receiving
Water Quality Criterion Based on the 48-hour Pacific
Oyster (Crassostrea gigas) Embryo. Washington
Dept. Fish. Tech. Rept. WDFTA7, No. 9, pp. 1-36.
Field Toxicity Testing Procedures
Jeffrey Black
I will be discussing some background and techniques
concerning field testing used both on site and in situ.
In the amount of time allotted, I will not be able to
cover all the methodologies used. However, I have
provided a rather extensive bibliography if more detail
is required. I've put some special emphasis on the
embryo-larval survival and teratogenicity test, because
Steve Schimmel asked me to describe that procedure
as it is used in both the laboratory and the field.
For purposes of discussion, the definition of "on site"
is testing performed in the field using mobile facilities
- in other words, bringing the lab to the field site.
In situ tests are those conducted instream.
Toxicity testing vs. chemical monitoring
There are advantages to performing ambient toxicity
testing as opposed to collecting only chemical moni-
toring data. My preference is to combine both
chemical monitoring and biomonitoring when possi-
ble, so that if toxicity is demonstrated the cause may
be discernible. But cause-effect relationships are not
always easily established. To choose between the two
kinds of monitoring, certain aspects of each must be
considered.
First, we must consider that water quality criteria
have been established for only a few chemicals
present in the environment. Second, the chemical
approach alone likely will not detect all the chemicals
in a contaminated system. Although chemical moni-
toring is appropriate when only a few chemicals are
known to be present in a system, it does not suffice
when many chemicals are present, especially consid-
ering possible synergistic or additive toxicant interac-
tions. In these situations we use organisms to "see"
those chemicals and chemical interactions in the
effluent or in the receiving water.
33
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We can make assessments of bioavailability, temporal
variability, and persistence of toxicity by using
biomonitoring. For determining the presence of
bioaccumulative chemicals, both chemical monitoring
and toxicological monitoring are appropriate, especial-
ly if the term "toxicological" can be extended to
encompass bioaccumulation studies. For rapid
assessments, chemical monitoring is probably more
efficient for detecting carcinogens, although some
pathologists might disagree, arguing that we can look
at indigenous aquatic organisms for tumor formation
and other biomarkers. Both chemical and toxicity
assessments are reliable for design of treatment
systems (e.g., chlorine reduction). In selecting
methods, of course expense must be considered; you
have to weigh what you want to do against the
availability of funds.
Purposes of testing
The purposes of ambient testing vary; different tests
are used to accomplish different objectives.
Some can be used for regulatory purposes. Many
times an industry's discharge permit is based on
characteristics of the mixing zone of the stream.
Ambient toxicity testing could be used to deter-
mine compliance in the mixing zone.
We may want to develop a scientific data base so
the findings of toxicity tests can be correlated with
ecological data to validate biomonitoring end-
points.
Ambient tests may be used to determine sources
of impact. For industries with only one or two
discharges, the sources are obvious. For nonpoint
sources such as agricultural runoff, ambient testing
can be useful in tracking important toxic inputs.
We can also assess temporal variability and the
persistence of toxicity from a source (spatial vari-
ability).
We can evaluate the effects of water quality
characteristics (e.g., salinity, hardness, pH) on
toxicity.
Selection of field stations
The selection of field stations includes a number of
important considerations. First of all, choosing a
station using map siting alone is usually unsatisfacto-
ry, as it does not indicate many limitations of access.
The proximity of stations to impact sources is also
very important in attempting to determine if there is
any spatial variability in effect. The establishment of
a control or reference site can be difficult. For in-
stance, an upstream site may be contaminated to the
extent that it is unsuitable. Thus we may be restricted
to using lab control water (reconstituted water). The
sites should be evaluated as to their importance as
spawning areas or recreational areas, and for their
potential for producing human health effects. For
field validation studies using benthic invertebrates,
fish populations, periphyton, etc., sites for ambient
testing should be selected close to the ecological
sampling sites.
The types of tests to be used — acute, chronic, or
"short-term" chronic — must be determined. In many
cases dealing with ambient toxicity, acute tests are not
adequately sensitive. Whether instream chambers or
mobile labs are used, the sampling procedures,
selection of test organisms, cost, and personnel needs
are all important, and these considerations have been
addressed by earlier speakers.
Variations in toxicity between lab and on-site evalua-
tions of effluents often have been observed. General-
ly, lab tests on stored effluents are less sensitive, in
some instances by up to two orders of magnitude
(Birge et al. 1985). In such cases, field testing with
a mobile facility has advantages over lab tests.
Standardized on-site test procedures vary. Acute tests
have been well documented in Methods for Measuring
34
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the Acute Toxicity of Effluents to Freshwater and
Marine Organisms (Peltier and Weber, 1985).
Chronic short-term tests for ambient waters thus far
validated to some extent are the fish embryo-larval
survival and teratogenicity test, fathead minnow larval
survival and growth test, and Ceriodaphnia survival
and reproduction test (Birge et al., 1989; Mount et al.,
1985, 1986).
Endpoints
As new tests are developed, reliable and reproducible
test endpoints must be established. The comparability
of values from short-term tests to results of longer-
term tests must be addressed. We compared LQ
(threshold) values with fish species to MATC values
for similar species in longer chronic tests and found
them to correspond in many cases (Birge et al., 1985).
In embryo-larval tests, teratogenicity is incorporated
with mortalities in calculating NOEC, LOEC, and LC
values, and these tests are applicable to both freshwa-
ter and saltwater species.
Several studies show that data from ambient toxicity
tests correlate to ecological stream parameters. More
work is needed in this area. If we don't know what
the lexicological endpoints mean in terms of the envi-
ronmental impacts, what good are they? So we have
compared embryo-larval test results with other ecolog-
ical parameters. We have found highly toxic situa-
tions in which receiving water killed all the test
organisms. We noted graded responses downstream
to a point where the survival range in the toxicity
tests was comparable to that observed for the control
area water. Studies revealed that the number of
macroinvertebrate taxa also correlated well with
embryo-larval survival frequencies.
Similar tests in mobile labs have incorporated several
marine species (Schimmel et al., 1989). Reproduction
of the red algae was the most sensitive endpoint, and
the survival and growth of sheepshead minnow larvae
were the least sensitive. Observed effects were
attributed to ammonia from a pulp and paper mill.
Instream evaluations include:
Juvenile fathead minnows (-250-350) in chambers
were used to study bioaccumulation of PCBs in
the Hudson River (Jones and Sloane, 1989).
In the upper Chesapeake and the Choptank River,
caged striped bass larvae and yearlings were
deployed from rafts, and composite chemical
samples were analyzed in conjunction with obser-
vations of bass mortality (Hall et al., 1988).
Using shorter-term tests, Borthwick et al. (1985)
and Clark et al. (1987) studied pink shrimp after
the field application of a mosquitocide (fenthion).
Mortality was observed within a 24-hour period,
and field observations confirmed effects seen in
laboratory studies.
A longer-term test was performed by Freeman and
Sangalang (1985) using the Atlantic salmon over
a 3-month period, with sampling conducted every
2-3 weeks for mortality and weight determina-
tions. The fish were collected at sexual matura-
tion and the differences between fish in acidic vs.
non-acidic conditions were determined. Fish
taken from acidic areas gained less weight, pro-
duced smaller eggs, and showed abnormal metab-
olism of steroid hormones.
Community toxicity tests
Community Toxicity Testing (ed. Cairns, 1986) is
a good reference (e.g., see Lewis et al., 1986).
Clements et al. (1988a) used artificial substrates
for studying aquatic insect communities. Sub-
strates were placed instream where ambient
toxicity was expected. Diversity and density were
measured after 30 days.
35
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Lewis et al. (1986) looked at the phytoplankton
and periphyton in stream and lake systems, adding
a toxicant back to the natural system. This proce-
dure is somewhat different from the type of
testing proposed for the Bay, but certain adapta-
tions to this method may be useful.
We used periphyton samplers (microscope slides
placed on a floating tray) in ambient water and
compared impacted areas for differences in density
and diversity. Sometimes we had major problems
when flooding removed many of our samplers, and
data collection was severely hampered. However,
these techniques can be very efficient in ascertaining
ambient water quality.
Problems
Other problems relate to situations when the water
taken from reference or control areas is toxic. Also,
we can change the integrity of samples when collect-
ing for on-site testing. For example, using standard-
ized procedures, we are generally limited to control-
ling temperature. Thus if temperature is a problem in
the ambient system, the standardized tests can't
provide this information. In this case, in situ proce-
dures may be more appropriate.
We need to return to examining the effects of natural
perturbations. Conventional pollutants must be as-
sessed, including suspended solids, chlorine, pH, and
oil and grease. With multiple discharges, il is diffi-
cult to assess cause-effect relationships in ambient
systems where toxicity is low and only occasionally
observed. Furthermore, we must evaluate the ecologi-
cal importance of toxicity endpoints. Statistical
procedures for interpreting results exist in manuals for
traditional types of toxicity testing, which use a
graded concentration range and "typical" controls.
How valid are these types of standardized statistics in
determining the significance of ambient toxicity?
Should toxicity in ambient systems be compared to
reconstituted control water? Does statistical signifi-
cance equal ecological significance?
What happens when you conduct a battery of tests -
two showing toxicity problems, two showing no prob-
lems, and two that don't work? This type of situation
can often occur, when data from test batteries are
conflicting.
I have no argument with looking at more than one
species, but the organisms must be carefully selected
and their importance validated. Research should
focus on more critical comparisons of test endpoints
to actual ecological effects. We also need to reassess
existing data and re-evaluate raw data from published
validation studies. Perhaps incorporating a combina-
tion of standardized and caged-organism testing would
be useful. I think we need not try to protect our own
turf any more, but rather talk with each other and de-
velop and integrate more approaches to ambient
toxicity testing.
Questions
Q:
A:
Have you used a reference toxicant with
a known effect (LC50)?
In response to the reference toxicant, yes,
we routinely use cadmium to assess vari-
ability of the responses of organisms and
the quality (viability) of the organisms.
The reference test with a chemical of
known toxicity is an important aspect of
field work, especially when using stan-
dardized procedures.
Comment: A cautionary note concerning the caged
test: it really depends on the individual species as to
the variance of metal response and uptake. We did
an acid-mine drainage-site study on caged trout. The
trout looked fine on the superficial level, but when we
looked at the subtle things like metal metabolism at
36
-------
the cellular, subcellular, and tissue levels, the caged
organisms, whether at control or impact sites, had
very different responses. The responses varied
particularly in uptake of metals, where the metals
were deposited, and susceptibility to metal toxicity.
Other species of fish were not very sensitive to it.
We must be careful in interpreting in situ tests,
because some species really don't like being caged.
Response: These considerations are important relative
to assessing toxicant-related vs. testing-related re-
sponses.
Q: How far apart are your testing stations?
A: It depends. In one study, we were deal-
ing with more than 40 miles of stream
from the effluent source to the last ambi-
ent station. In other studies, the distance
was much less. Station locations are
specific to the study site.
Birge, W.J., J.A. Black, and A.G. Westerman. 1985.
Short-term fish and amphibian embryo-larval tests for
determining the effects of toxicant stress on early life
stages and estimating chronic values for single com-
pounds and complex effluents. Environ. Toxicol.
Chem. 4: 807-821.
Black, J.A. and W.J. Birge. The fish embryo-larval
procedure: predicting chronic toxicity and ecological
effects. Fish Physiology, Fish Toxicology, and
Fisheries Management, International Symposium
Proceedings, Guangzhou, PRC. (R.C. Ryans, ed.). In
Press.
Borthwick, B.W., J.R. Clark, R.M. Montgomery, J.M.
Patrick, Jr., and E.M. Lores. 1985. Field confirmation
of a laboratory-derived hazard assessment of the acute
toxicity of fenthion to pink shrimp, Penaeus duora-
rum. Aquatic Toxicology and Hazard Assessment:
Eighth Symposium. ASTM STP 891, R.C. Bahner
and D.J. Hansen, Eds., ASTM, Philadelphia, pp. 177-
189.
References
Birge, W.J. and J.A. Black. 1981. In situ
acute/chronic lexicological monitoring of industrial
effluents for the NPDES biomonitoring program using
fish and amphibian embryo-larval stages as test
organisms. OWEP-82-001. U.S. EPA. Washington,
DC.
Birge, W.J. and J.A. Black. In situ lexicological
monitoring: use in quantifying ecological effects of
loxic wasles. In Situ Evalualion of Biological Haz-
ards of Environmental Pollutants (S. Sandhu, ed.).
Plenum Publishing Co., NY. In Press.
Birge, W.J., J.A. Black, T.M. Short, and A.G. Wester-
man. 1989. A comparative ecological and toxicolog-
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Clark, J.R., B.W. Borthwick, L.R. Goodman, J.M.
Patrick, Jr., E.M. Lores, and J.C. Moore. 1987.
Comparison of laboratory toxicity test results with re-
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the field. Environ. Toxicol. Chem. 6: 151-160.
Clements, W.H., D.S. Cherry, and J. Cairns, Jr.
1988a. Structural alterations in aquatic insect com-
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Environ. Toxicol. Chem. 7: 715-722.
Clements, W.H., D.S. Cherry, and J. Cairns, Jr.
1988b. Impact of heavy metals on insect communi-
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Dawson, D.A., C.A. McCormick, and JA. Bantle.
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sis assay - Xenopus (FETAX). J. Appl. Toxicol. 5:
234-244.
DeGraeve, G.M. and J.D. Cooney. 1987. Ceriodaph-
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col. Chem. 6: 331-333.
Freeman, H.C. and G.B. Sangalang. 1985. The
effects of an acidic river, caused by acidic rain, on
weight gain, steroidogenesis, and reproduction in the
Atlantic salmon (Salmo salar). Aquatic Toxicology
and Hazard Assessment: Eighth Symposium. ASTM
STP 891, R.C. Banner and D.J. Hansen, Eds., ASTM,
Philadelphia, pp. 339-349.
Hall, L.W., Jr., SJ. Bushong, M.C. Ziegenfuss, W.S.
Hall, and R.L. Herman. 1988. Concurrent mobile on-
site and in situ striped bass contaminant and water
quality studies in the Choptank River and Upper
Che&2sapeake Bay. Environ. Toxicol. Chem. 7: 815-
830.
Toxicity Testing, ASTM STP 920, John Cairns, Jr.,
Ed., ASTM, Philadelphia, pp. 224-240.
Mount, D.I. and T.J. Norberg. 1984. A seven-day
life-cycle cladoceran toxicity test. Environ. Toxicol.
Chem. 3: 425-434.
Mount, D.I., T.J. Norberg-King, and A.E. Steen.
1986. Validity of effluent and ambient toxicity tests
for predicting biological impact, Naugatuck River,
Waterbury, Connecticut. EPA/600/8-86/005. U.S.
EPA, Duluth, MM.
Mount, D.I., A.E. Steen, and T.J. Norberg-King.
1985. Validity of effluent and ambient toxicity
testing for predicting biological impact on Five Mile
Creek, Birmingham, Alabama. EPA/600/8-85/015.
U.S. EPA. Duluth, MN.
Norberg, T.J. and D.I. Mount. 1985. A new fathead
minnow (Pimephales promelas) subchronic toxicity
test. Environ. Toxicol. Chem. 4: 711-718.
Jones, P.A. and R.J. Sloan. 1989. An in situ river
exposure vessel for bioaccumulation studies with
juvenile fish. Environ. Toxicol. Chem. 8: 151-155.
Knight, J.T. and W.T. Waller. 1987. Incorporating
Daphnia magna into the seven-day Ceriodaphnia
effluent toxicity test method. Environ. Toxicol.
Chem. 6: 635-645.
Lewis, M.A., M.J. Taylor, and R.J. Larson. 1986.
Structural and functional response of natural phyto-
plankton and periphyton communities to a cationic
surfactant with consideration on environmental fate.
Community Toxicity Testing, ASTM STP 920, John
Cairns, Jr., Ed,, ASTM, Philadelphia, pp. 241-268.
McCormick, P.V., J.R. Pratt, and J. Cairns, Jr. 1986.
Effect of 3 trifluoromethyl-4-nitrophenol on the
structure and function of protozoan communities
established on artificial substrates. Community
Peltier, W.H. and C.I. Weber. 1985. Methods for
measuring the acute toxicity of effluents to freshwater
and marine organisms. EPA 600/4-85-013. U.S. EPA.
Cincinnati, OH.
Sayre, P.O., D.M. Spoon, and D.G. Loveland. 1986.
Use of Heliophyra , a sessile suctorian protozoan, as
a biomonitor of urban runoff. Aquatic Toxicology
and Environmental Fate: Ninth Volume, ASTM STP
921, T.M. Boston and R. Purdy, Eds., ASTM, Phila-
delphia, pp. 135-153.
Schimmel, S.C., G.B. Thursby, MA. Heber, and M.J.
Chammas. 1989. Case study of a marine discharge:
comparison of effluent and receiving water toxicity.
Aquatic Toxicology and Environmental Fate: Eleventh
Volume, ASTM STP 1007, G.W. Suter II and M.A.
Lewis, Eds., ASTM, Philadelphia, pp. 159-173.
38
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Tagatz, M.E. 1986. Some methods for measuring dubia toxicity tests for cadmium and sodium penta-
effects of toxicants on laboratory- and field-colonized chlorophenate. Environ. Toxicol. Chem. 7: 153-159.
estuarine benthic communities. Community Toxicity
Testing, ASTM ATP 920, John Cairns, Jr., Ed.,
ASTM, Philadelphia, pp. 18-29.
U.S. EPA. 1985. Technical support document for
water quality based toxics control. Office of Water,
Washington, DC.
Wang, W. 1987. Toxicity of nickel to common
duckweed (Lemna minor). Environ. Toxicol. Chem.
6: 961-967.
Wang, W. and J.M. Williams. 1988. Screening and
biomonitoring of industrial effluents using phytotoxic-
ity tests. Environ. Toxicol. Chem. 7: 645-652.
Warren-Hicks, W, B.R. Parkhurst, S.S. Baker, Jr.
1989. Ecological assessment of hazardous waste sites:
a field and laboratory reference. EPA/600/3-89/013.
U.S. EPA. Corvaffis, OR.
Weber, C.I. 1973. Biological field and laboratory
methods for measuring the quality of surface waters
and effluents. EPA-670/4-73-001, U.S. EPA. Cincin-
nati, OH.
Weber, C.I. et al., 1989. Short-term methods for
estimating the chronic toxicity of effluents and
receiving waters to freshwater organisms. EPA-
600/4-89-001. U.S. EPA. Cincinnati, OH.
Weber, C.I., W.B. Horning II, D.J. Klemm, T.W.
Neiheisel, P.A. Lewis, E.L. Robinson, J. Menkedick,
2nd F. Kessler. 1988. Short-term methods for esti-
mating the chronic toxicity of effluents and receiving
waters to marine and estuarine organisms. EPA-
600/4-87/028. U.S. EPA. Cincinnati, OH.
Winner, R.W. 1988. Evaluation of the relative
sensitivities of 7-d Daphnla magna and Ceriodaphnia
39
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Plenary Session D:
Methodologies for Suborganismal (Biochemical and Cellular)
Toxicity Testing
Conveners: Jay W. Gooch, Peter Van Veld
Gooch: I appreciate Dr. Black's suggestion that we
need to break down some of the barriers in our
thinking between ecological endpoints and subcellular
endpoints. I think that those of us studying the
subcellular level do have a contribution to make to
the overall discussion.
For this afternoon's presentations, Dr. Roesijadi will
begin with more of a philosophical discussion of
biochemical endpoints; where they've come from,
expectations about the information they will provide,
and limitations to linking the endpoints to population
or community level impacts. I will follow up with
more detailed information about some of the potential
measures and demonstrate the kind of information
they can provide.
Rationale for and Relevance of Analysis of
Suborganismal Responses to Contaminant
Exposure
G. Roesijadi
A major question regarding the analysis of suborgan-
ismal responses to contaminant exposure is "Why has
an approach with the power and ability to solve
problems in human health not been readily adopted to
solve problems of environmental health?" If we
understand Suborganismal responses — both the
normal functions and deviations caused by pathologi-
cal conditions - we can use that information to deal
with problems related to the exposure of organisms in
the environment. Understanding the mechanisms of
toxic action can provide tools for detecting and
assessing a toxicant's effects. However, the general
premise of such a clinical approach to assessing
environmental health has not yet been accepted.
The recent history of this field goes back some 15-20
years when biologists began to actively pursue a
greater understanding of the mechanisms underlying
the effects of environmental contaminants in aquatic
organisms. The concept of a diagnostic approach was
adopted and has been variously referred to as suble-
thal effects, biochemical indicator responses, biologi-
cal effects monitoring, and, currently, biomarkers.
These synonyms reflect attempts to establish an
identity for a developing field of environmental
science.
General Framework for Biological Effects
At the present time, we know a good bit about
measuring pollutant inputs and dispersive processes
(Figure 1).
Figure 1.
41
-------
In the realm of analysis of environmental processes,
we are able to deal relatively well with describing the
chemical environment and bioaccumulation by organ-
isms. Conceptual frameworks and analytical proce-
dures associated with measuring these processes are
generally well-accepted. We do not have a good
understanding of the means for assessing biological
effects. If we did we would not have convened a
forum such as this in which the merits of design of
assessment and monitoring programs are being
discussed. We are here because we need to assess
approaches to take and need to come to a consensus
on which available ones will be productive.
By suborganismal responses, we mean responses that
are mainly at the cellular and subcellular levels of
biological organization (Figure 2).
Flgur* 2.
BIOLOGICAL LEVELS
OF ORGANIZATION
SUBCELLULAR
LYSOSOMAL LATENCY
ENZYME ACTIVITY
METABOLIC FLUXES
ENERGY CHARGE
BLOOD CHEMISTRY
DMA AND CHROMOSOMAL
ABNORMALITIES
METALLOTHIONEINS
MIXED FUNCTION
OXYGENASES
ORGANISING
(Physiological)
RESPIRATION
FEEDING
EXCRETION
GROWTH
FECUNDITY
REPRODUCTIVE EFFORT
LARVAL VIABILITY
CELLULAR/TISSUE
QAMETOQENIC CYCLE
NUTRIENT STORAGt CYCLE
DEFORMITY
NEOPLASMS/TUMOURS
POPULATION
BIOMASS
PRODUCTIVITY
AGE/SIZE STRUCTURE
RECRUITMENT
MORTALITY
COMMUNITY
BIOMASS
ABUNDANCE
DIVERSITY
TROPHIC INTERACTIONS
ENERGY FLOW
VARIABILITY
RESPONSE TIME
SENSITIVITY
QUANTITATIVE
SPECIFICITY
SIGNAL:NOISE
MONTHS TO YEARS
'DAYS TO WEEKS
•-»-YEAFIS TO DECADES
HK3H
•LOW
HIGH
--»• LOW
SPECIFIC/GENERAL
HIGH
ECOLOGICAL
RELEVANCE HIGH
-*• GENERAL
——f LOW
If one looks at biological levels of organization from
the subcellular to the community and examines
parameters such as response time, sensitivity, ability
to quantify, specificity, signal-to-noise ratio, and
ecological relevance, the more detailed levels of
organization can provide a higher degree of rapid,
sensitive, and quantifiable information. However, the
ecological significance of such measurements is
considered low. At the more complex levels of
organization where ecological relevance is high, the
picture becomes blurred as signal-to-noise becomes
less evident and the time-scales required to obtain
information are long. One of the difficulties in the
study of suborganismal responses is the problem of
relating information derived at lower levels of biologi-
cal organization to more complex levels; in other
words, the ability to extrapolate effects from lower to
higher levels of organization.
Ecological Relevance of Suborganismal Responses
In the past, we have tended to view all levels of
biological organization in a nested fashion (Figure 3
top).
Rgur* 3. Perceived relationships of various categories of
biological organization (top figure from Molecular Ecology
Institute, California State University, Long Beach).
Strict adherence to this view presents a problem,
because it implies that processes at molecular and
cellular levels can be directly extrapolated to popula-
tions and communities. There is a demand that
measurements of cellular processes be extrapolated to
population- and community-level processes and thus
42
-------
have ecological significance. I think that processes
measured at the suborganismal levels are ultimately
useful in explaining responses at the individual level.
They can reflect on the population in so far as the
measurements being made are representative of
responses of individuals in a given population.
Extrapolation of such measures to population process-
es is debatable. Thus, another representation of the
relationship between the various levels of organization
is presented in Figure 3 (bottom) in which the subor-
ganismal and organismal processes are less directly
coupled to the population and community, although a
relationship is recognized. We have to ask whether
processes such as reproductive output, which is
considered to be an ecologically-relevant measure, are
directly coupled to recruitment, which is a population
parameter. The relationship may not be direct.
Recruitment processes are not well enough understood
for us to make unequivocal statements about the
relationship between reproduction by individual
organisms and population responses. Arguments that
relate to density-dependent and -independent controls
would also be applicable. If we understand processes
at the more detailed levels of biological organization
represented by suborganismal responses and have
enough information on their relationship to organismal
function, we can extrapolate to processes that ulti-
mately relate to organismal functions such as growth
and reproduction. Other factors, referred to as emer-
gent properties by some, may then control how the
individual relates to the population and community.
10"
10"
10"
I
10'
to'
10'
10'
HDEKHOtNT
10 JO
TME (DAYS)
100 tOD MS tOO
Figure 4. A conceptualization of factors that influence the
abundance of fishes during the recruitment process (from E.
Houdc, 1987, Fish e«rty life dynamics and recruitment
variability, Am. Fish. Soc. Symp. 2:17-29).
A figure on recruitment (Figure 4) from a paper by
Dr. Edward Houde shows that many factors can influ-
ence the actual numbers recruited into a fish popula-
tion. He has identified the period of larval develop-
ment as one that is most sensitive to changes in
environmental parameters and other factors such as
predation and food availability.
Figure S.
HYPOTHETICAL RECRUITMENT OF YOUNG FISH UNDER
ONE GOOD AND THREE BAD CONDITIONS (*25% CHANGE
IN MORTALITY OR GROWTH RATES) (HOUDE. 1987)
AGE AT
CONDITION INITIAL n MOFT[A'JTV METAMORPHOSIS RECRUITED n
*""' (DAYS)
QOOO 1x10*
BAD-1 1x10*
BAD-2 1x10*
BAD-3 1x10*
0.100
0.125
0.100
0.125
45.0
45.0
56.2
56.2
11,106
3.BO7
3,625
889
FIGURE 5 TABLE ON HYPOTHETICAL RECRUITMENT OF LARVAL FISH
UNDER VARIOUS CONDITIONS AFFECTING GROWTH AND SURVIVAL (FROM
E. HOUDE, 1887, FISH EARLY LIFE DYNAMICS AND RECRUITMENT
VIABILITY, AM. FISH SOC. SYM. 2:17-29).
In a table of hypothetical recruitment (Figure 5), he
gives four examples that include one with "good"
recruitment conditions and three with "bad." The
three bad conditions were a 25% increase in mortali-
ty, 25% increase in age-at-metamorphosis (a reflection
of growth rate), and both simultaneously. These
"bad" conditions can occur independently of any
pollution-related processes and cause significant re-
ductions in numbers recruited. If the "Initial n" in the
table is altered to simulate changes in reproductive
output, using the same values for mortality and age-
at-metamorphosis as in the good condition, very large
changes in "Initial n" would have to occur to cause
reductions in recruitment similar to the "bad" condi-
tions. Thus, to obtain reductions in recruitment
equivalent to "bad" conditions 1 and 2, a 68% reduc-
tion in "Initial n" is needed. To achieve reductions
equivalent to "bad" condition 3, a 92% reduction in
"Initial n" is needed. The effects of such reductions
as a result of contaminant-related effects may not be
detectable in the context of natural variability in
recruitment. Coupling responses at the individual
43
-------
level with population processes can be extremely
difficult. Demonstrated effects on "ecologically-
meaningful" measures such as growth (of inappropri-
ate life stages) and reproduction in an experimental
context may have little meaning in the real world.
If we can accept the premise that it is very difficult to
couple lower and higher level processes directly as
one goes from the individual to the population, then
suborganismal responses will have a different frame
of reference and take on new meaning. It must be
recognized that the demand for extrapolation to
population processes will most likely not be met when
using suborganismal responses. Therefore, the value
of responses of individuals would have to carry more
weight in environmental assessment than they current-
ly receive. Diagnostic and clinical approaches would
then increase in relevance, having meaning at the
individual level. Proper diagnosis and appropriate
sampling procedures can provide measures of the
presence and extent of adverse change within existing
members of a population. This information in itself
should be of value for those needing to make immedi-
ate decisions relating to environmental contamination
and its ramifications. The limitation of uncertain
ecological extrapolation should not deter us from
using an approach with the greatest potential for early
and sensitive detection of contaminant-related effects.
As in the detection and diagnosis of sick humans by
the medical profession, there should be intrinsic value
in the detection and diagnosis of sick animals in the
natural environment.
Summary
The knowledge that individuals are adversely
affected by exposure has intrinsic value and
should be used in environmental management.
The expectation of extrapolation of measures
based on individuals to ecologically-significant
processes is currently unrealistic.
Diagnostic approaches known to be effective in
epidemiology and medicine will be effective in
environmental toxicology.
However, to effectively utilize such approaches,
we will have to reevaluate the value system that
assumes that effects must have demonstrated
ecological significance for them to be valid mea-
sures of biological effects and suitable for man-
agement decisions.
Questions
Comment: It is difficult to go from organism to
population, but it is important to understand that much
of what we do is based on the individual organism.
Comment: I disagree. It is not true that populations
are uncoupled from individuals. Populations of some
species (r-selected) are driven by environmental
conditions, but many (including some that society is
particularly concerned about, like striped bass) are
not. A 68% reduction of reproduction rates is not
high.
Response: But this is about the same level of reduc-
tion that was produced by changes in mortality or
longevity of larval stage; this was the point. Natural
changes can be too small to detect but make very
large changes in recruitment. The other point is that
matters like coupling are highly controversial in the
ecological field.
Comment: No more controversial than the prediction
of individual responses from biomarkers.
Response: The point was that in dealing with organis-
mal responses, the demand of extrapolating to popula-
tion responses may be unrealistic. "Biomarkers" is
for me a red flag. I think we can accept that cellular
processes do affect fitness of individuals - we accept
it in humans, anyway. The connection may not be
44
-------
well demonstrated in the species we study, but that
doesn't mean it is nonexistent.
Comment: The linkage between individual and
population responses can be variable, depending on
environmental/ecological factors, which may be
stronger in some years than in others. It has to be
viewed in the context of the natural environment.In
doing some preliminary scope-of-growth studies it
occurred to me that the fecundity level shows the
energy level built up over a few weeks, and some of
these tissue level tests can be done over a couple of
days. So we see some integrative function from
subcellular tests, and some things that may be early
warnings. The problem is quantifying what finally
happens to the animals. The quantitative links must
be established by future research.
Techniques for Assessing the Sublethal
Effects of Chemical Contaminants on
Aquatic Life
Jay W. Gooch
The use of biomarkers and their integration into
routine testing schemes has so far met with resistance.
The regulatory and management communities don't
necessarily understand how this approach helps them.
It is the responsibility of the research community to
make the importance of this level of information
apparent and to communicate realistic expectations
about the information provided. This is an emerging
technique, and the question is whether this extra
information gives us anything we need to know.
One of the original justifications for the use of
sublethal endpoints in environmental testing was the
desire to have an early warning system. It was
recognized that by the time resource managers could
detect declines in populations, the damage to the
resource had already occurred.
One of the problems we encounter in the use of
biochemical endpoints in environmental research is
the emphasis on population or community level
responses. These measurements usually come into
use through progress that is made in mammalian
research where we place an intrinsic value on the
individual response, largely because we are trying to
protect the health of individual humans. By contrast,
in environmental research we do not place our empha-
sis on the individual; hence, there is a resistance to
studies focused at this level.
I have listed a number of the justifications that have
been put forth for the use of biochemical endpoints.
These endpoints:
• assess the "health" of animals, serving as diag-
nostic tools.
• provide early warning of habitat degradation.
• integrate multiple exposures and interactions.
• account for modifiers of toxicity.
• can serve as measures of exposure.
• can be useful in tracking environmental recov-
ery.
One of the initial issues to discuss is specific vs.
general measures. In the literature you will find that
there is no general agreement about this issue, and
there is not enough time here to discuss the arguments
in great detail. Rather, I will discuss some of the
advantages and disadvantages of each. Clearly,
depending on the circumstances and the questions
being asked, there may be merit to one approach or
the other, or some combination. In general, I don't
think anyone would suggest that there is any one
magical biomarker that will provide information under
all circumstances.
45
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Specific vs. general measures
Specific tests provide a strong mechanistic interpreta-
tion, i.e., links between cause and effect. Responses
can be grouped by compound class as related to mode
of action. One of the problems, however, is that
information is lacking for some groups of compounds,
or the mode of action leaves unknown changes at
sublethal levels (e.g. cyclodiene insecticides).
Another argument in favor of specific measures is
that there is a large mammalian data base to draw
from. As I stated earlier, many of the measures we
use have evolved out of studies done in mammalian
systems, where the mechanistic approach to toxicolo-
gy prevails and consequently these measures are
generally more developed.
Some examples of specific measures are induction of
cytochrome P450-mediated monooxygenase enzymes
by certain organic pollutants, induction of metallo-
thioneins (metal-binding proteins) by metals;, inhibi-
tion of acetylcholinesterase by organophosphate and
carbamate insecticides, and inhibition of delta amino-
levulinic acid dehydrase by lead. Clearly, this is not
an all-inclusive list.
The greatest strength of general responses is that they
are ideally sensitive to a wider group of stressors.
However, we usually have less background informa-
tion on "normal" ranges for these parameters and
understand less about how they are affected by natural
environmental variables. In addition, there is less
mechanistic interpretation or cause/effect linkage
possible. For a manager this means that the measure
by itself will not be able to suggest any particular
contaminant. More information would be needed in
order to determine where regulation or enforcement
should be enhanced. I am not suggesting that specific
measures provide all of the information that is needed
either. The point is that the information gained is
directly related to the tools that are used.
Below I have listed a number of different general
measures of response. My bias is to lump many of
the measures that others consider more whole-organ-
ism/physiological measures into this category of
general measures of response. I won't discuss any of
these in great detail as time does not permit. Recog-
nize that this is only a partial list. A good summary
of many of the enzymes or measures that have been
used can be found in Neff (1985).
Examples:
RNA/DNA ratios
adenylate energy charge
glycogen levels
scope for growth
amino acid profiles
serum chemistry (important in mammalian sys-
tems)
histopathology
others
Because the preliminary documents that were distrib-
uted contained a list of some of the candidate mea-
sures for this effort, I tended to focus on them in my
preparation. Again, this is not intended to be an all-
inclusive list.
Candidate measures
I would like to preface this part of my talk to reveal
my bias up-front. The cytochrome P450 enzymes are
one of my research areas. I am going to dwell on
them somewhat, not to sell you on them as the only
relevant parameter to measure, but rather to use them
as an example of the kinds of data that are being
46
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generated from field studies using subcellular mea-
sures. Also, I think they represent a useful example
of the kinds of things that need to be considered when
determining the significance of these kinds of mea-
sures.
Toxification and detoxification enzymes. In the
general reaction scheme of the primary and secondary
biotransformation pathways, the first reaction is often
referred to as a phase I transformation. This is the
initial reaction required to transform a lipophilic
molecule to one that is more water soluble, or to alter
the molecule so that it can be further metabolized to
a water-soluble form by secondary or phase II reac-
tions. The first reaction is catalyzed by the cyto-
chrome P 450-mediated monooxygenase system. One
view suggests that this family of enzymes has evolved
in response to the need to protect the body against
potentially harmful foreign chemicals, particularly
those entering via the diet. The attribute of this
system that provides a useful measure in an environ-
mental context is that specific forms of P450 enzymes
(detected as activity or protein) are induced by
chemical exposure. In other words, when animals are
treated with various chemicals, the transcriptional
machinery of the cell, particularly the liver cell, is
activated, and increased amounts of specific forms of
P450 enzymes are synthesized. It is this enzyme
induction, as it is referred to, that provides a specific
measure of exposure.
Without going into too much detail, let me just tell
you that fish respond in a much more limited way
than do mammals. Kleinow et al. (1987) list a
number of the fish species that have been investigated
and the compounds that they have shown a response
to. The important thing to note is that fish P450
enzymes respond consistently only to the planar, and
often balogenated, chemical contaminants. Fortunate-
ly or unfortunately, depending on your point of view,
these compounds are some of the more ubiquitious
and toxic pollutants we have in aquatic systems.
These pollutants are the PCBs, PBBs, PAHs, dioxins,
dibenzofurans and the aromatic fraction of petroleum.
There are several reasons why the functional status of
this system may be important to an animal. For a
number of compounds, metabolism is an activation
process, i.e., the metabolic product is more toxic than
the parent compound. In addition, many metabolites,
particularly those of PAHs, are carcinogenic. Hence,
the status of P450 enzymes may play a determinant
role in the effects of a chemical on an organism. For
example, Dr. John Lech's laboratory (Erickson et al.,
1986), studied rotenone toxicity under various condi-
tions of the P450 system in rainbow trout. Under
normal (i.e., control) conditions, rotenone exhibits a
96-hour LC50 of approximately 3.5 parts per billion.
When the fish were treated with beta-naphthoflavone,
an inducer of P450 activity, rotenone was actually
less toxic, with an LC50 now near 4.5 parts per
billion. In addition, when fish were treated with
piperonyl butoxide, an inhibitor of P450-based metab-
olism, rotenone was considerably more toxic than in
untreated fish. This demonstrates that the status of
the P450 system, as affected by exposure to other
compounds, can affect toxicity in a subsequent
exposure.
This is one justification for looking at this suite of
enzymes in the context of environmental risk assess-
ment. A population that has elevated P450 enzymes
that are responsible for metabolic activation may be
at increased risk to subsequent exposure. This might
interface to uncertainty in a risk assessment.
Dr. John Stegeman and coworkers at the Woods Hole
Oceanographic Institution (Stegeman et al., 1986)
were able to detect significant increases in the major
aromatic hydrocarbon-inducible form of P450 using
an antibody probe, demonstrated in a western blot.
One band, representing the elevated level of this
protein in a deep-sea fish collected off the continental
shelf east of New York, was contrasted with another
band from a fish collected off the continental shelf
47
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east of Nova Scotia. Dr. John Farrington's laboratory
analyzed the livers from these fish and found that the
levels of PCBs were much higher in the fish collected
off New York. While we cannot say conclusively
that these are causally related, the association is
strong circumstantial evidence for a biochemical
impact of pollution even in the deep sea.
There have recently been several field-oriented
workshops on biological effects measurements. I
would like to use some of the data that was generated
during an International Oceanographic Commission
(IOC)-sponsored workshop conducted by its Group of
Experts on the Effects of Pollutants (GEEP) two years
ago in Norway. In this study, investigators examined
animals collected along a pollution gradient moving
up a Norwegian fjord (Bayne et al., 1988). The
practical workshop and the collection of papers that
resulted from it represents a good example of the kind
of study necessary to place subcellular measures in
context with physiological and ecological measures.
A comparison of the changes in cytochrome P450
monoxygenase enzymes in flatfish sampled from
different stations along the pollution gradient showed
a strong relationship between enzyme activity and the
degree of pollution with aromatic hydrocarbons. This
was compared with scope-for-growth measurements
that were made by another group for the mussel
Mytilus edulis. The scope for growth in the mussels
decreased with increasing pollution just as the P450s
in the flatfishes increased. This type of association
gives us confidence that subcellular measures can be
interpreted at higher levels.
An example of the relationship between subcellular
measures and reproduction can be found in work
done in the laboratory of Dr. Robert Spies at the
Lawrence Livermore Laboratory in California (Spies
et aL, 1988a, 1988b). His group has been studying
the starry flounder in San Francisco Bay and examin-
ing the relationship between cytochrome P450 activity
in spawning females and various parameters associat-
ed with the viability of the young. The correlation
between embryological success and P450 activity is
significant, but not particularly predictive. These
kinds of data are also provocative when attempts are
made to put subcellular measures in context with
ecological measures.
A promising measure which has received attention is
the level of metallothioneins. These proteins are
intimately involved in trace metal homeostasis and in
the metal balance of the cell. They can in fact be
protective. Studies have shown that when metal-
binding proteins have been induced in animals,
subsequent exposure often results in a lowered sensi-
tivity. In other words, naive animals have greater
sensitivity than those exposed to metals. Thus, the
presence of high levels of metallothioneins in animals
collected in the wild may be a signal of trace metal
exposure.
Immune alterations. Another intuitively attractive
measure is the functional status of the immune sys-
tem. Measures of the functional status of the immune
system and its compromise by chemical exposure
have been highlighted as one of the important areas of
suborganismal response that is poorly understood but
potentially very important. The presence and spread
of diseases like Dermo and MSX in shellfish stocks
of the Bay have heightened this awareness in this
region.
There are two types of measures that have been
suggested for looking at immune suppression. For a
direct measure, an organism can be challenged with
a pathogen and the clearance of the pathogen fol-
lowed. For an indirect measure, you can observe the
functional aspects of the immune system: antibody
production, phagocytic cell function, macrophage
aggregation, etc.
Stress protein induction. Stress proteins are found in
species from microbe to man and are rapidly synthe-
sized in response to acute stress. Examples of these
kinds of stress are heat (hence the original name heat-
48
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shock proteins), oxidizing agents, heavy metals,
steroid hormones, anoxia, low pH, viral infections,
wounding, antibiotics, etc. Studies conducted by Dr.
Brian Bradley on bivalves and copepods here in the
Chesapeake Bay show how the various molecular
weight proteins that are induced can be detected using
immunological probes. It has been shown that the
profile of the various heat-shock proteins induced may
provide information regarding the particular form of
"stress" incurred.
DNA alterations and genotoxicity. DNA alterations/-
genotoxicity show great promise in revealing underly-
ing factors in environmental carcinogenesis. They
have been used largely in the context of PAH contam-
ination and PAH-related lesions. As I stated earlier,
PAHs are metabolized to reactive intermediates by the
P450 system, a factor which may be related to the
presence of cancerous lesions in areas heavily pollut-
ed with PAH. In general, this approach looks at
changes in genetic structure, either in chromosomal
structure at different phases of the cell cycle or in
adducts that have formed from reactive species (e.g.,
the PAH metabolites).
Oncogene induction is one of the least explored
realms. This is a very active area of mammalian
cancer research. The approach is to look at the
inappropriate or unregulated expression of genes that
control cellular growth processes. It appears as
though these phenomena may play a major role in the
growth and proliferation of certain kinds of tumors.
While investigation of these in the context of tumors
in feral fishes is an important endeavor, it is unclear
whether they will have widespread application in
other forms of environmental testing. Studies on the
molecular expression of other growth factors, such as
growth hormone and thyroid hormones, however, may
prove to have applications.
The disposition of those of us concerned with bio-
chemical endpoints is that we hope to make measure-
ments with integrative and predictive power. We
aren't there yet, but some of these kinds of measure-
ments have promise and utility.
Most of these measures haven't been looked at in
standard toxicity testing protocols, but there is no
particular reason why they couldn't be. Many of
these measures could simply be added to the list of
measurements taken at the end of the test. The
problem is that people trained in one type of testing
aren't usually trained in the other. Some coming
together is needed.
On the subject of coming together, I want to explain
why the decision was made to cancel the workgroup
session on suborganismal reponses. This week in
Keystone, Colorado, SETAC, the Society of Environ-
mental Toxicology and Chemistry, is sponsoring a
workshop on biomarkers. Many of the people we had
hoped to have participate in the session here are going
to be at this meeting. I have the list of participants if
anyone is interested in seeing it. The format of that
meeting is to have seven working groups, six in
various topic areas and one synthesis group. There is
a group titled Physiology/Other being chaired by Dr.
Foster Mayer of the U.S. EPA; a group on Metabo-
lites being chaired by Dr. Mark Melancon, now with
the U.S. Fish and Wildlife Service in Patuxent; a
group on Histopathology being chaired by Dr. Dave
Hinton of the University of California-Davis; a group
addressing Protein, Enzyme Induction/Inhibition being
chaired by Dr. John Stegeman of the Woods Hole
Oceanographic Institution; a group addressing Immu-
nology being chaired by Dr. Beverly Ann Weeks of
VIMS; and a group addressing DNA Alterations being
chaired by Dr. Lee Shugart of the Oak Ridge National
Laboratory. Those of you familiar with this field will
recognize that a number of these people are the
experts and it was felt that the results and recommen-
dations put forth from this workshop would be
directly applicable to the Chesapeake Bay workplan.
I will make every effort to obtain and incorporate the
results of the SETAC workshop in a timely fashion.
49
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Questions
Q: Do you feel that in the regulatory context any
biomarker should be tied in some direct fashion
to growth, survival, or reproduction of an organ-
ism?
A: Not until we can show that link. With some of
these endpoints, if we can show direct links to
growth, we may have some scientific power and
credibility. It will take a brave regulator at
some point to regulate on the basis of these, and
it will get battled out in court -- if it ever hap-
pens.
Q: EPA has been bludgeoned when it tried to use
this kind of endpoint without proper basis. We
need concepts that are usable and understandable
by laymen and judges. The tie must be firm and
pronounced.
A: You're right, it won't be easy.
Comment: Uses for biomarkers are various. They
can be used, for instance, as an indicator of exposure
with preliminary biological results. We have to know
how we're using these and take advantage of them
where we can.
River, we targeted toadfish for cyto-
chrome P450 studies. Toadfish carry-
ing high levels of PAH metabolites in
bile showed no change in cytochrome
P450-mediated enzyme activity, in tests
done at three labs. In spot, responses
were what you'd expect. Choice of
species is crucial.
Q: Has anyone looked at Fundulus in chronically
stressed areas vs. nearby unstressed areas?
A: Certainly. This type of study is traditional.
Also, it seems that certain genotypes appear to
be selected for in these situations.
Q: What about the theory recently expressed in
Nature that extrapolation from rats to humans is
absurd?
A: We can't test humans; we have to have surro-
gates, so even if they aren't perfect these studies
have some value. At biochemical levels there are
problems of extrapolation between species. In
terms of toxicity, i.e., do they live or die, you
also see inter-species differences, and these are
often related to the same biochemical differenc-
es.
Q: Extrapolation across levels of organization has
been discussed. The other problem is extrapo-
lating across species. In a study of San Joaquin
kit foxes, we were doing blood samples for
standard veterinary blood diagnostics. The
response from the veterinary consultant looking
at the samples was that if they were dogs they
were dead. How do you deal with that kind of
problem, when you can't develop for every
species the kind of database we have for hu-
mans?
A: Yes, it's a problem sometimes even in ihe same
phyla. For example, in a cruise on the Elizabeth
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Neff, J.M. 1985. Use of biochemical measurements to
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Roesijadi, G. 1980. The significance of low molecular
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Shugart, L.R. 1988. Quantitation of chemically
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alkaline unwinding assay. Aquat. Toxicol. 13:43-52.
Spies, R.B, D.W. Rice, Jr. and J. Felton 1988a.
Effects of organic contaminants on reproduction of
the starry flounder Platichthys stellatus in San
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function oxidase (MFO) activity during the reproduc-
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Spies, R.B. and D.W. Rice, Jr. 1988b. Effects of
organic contaminants on reproduction of the starry
flounder Platichthys stellatus in San Francisco Bay.
II. Reproductive success of fish captured in San
Francisco Bay and spawned in the laboratory.
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Stegeman, J.J., P. J. Kloepper-Sams and J.W. Farring-
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Science 231:1287-1289.
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Final Plenary Session
Concluding Remarks and Workgroup Session Reports
Moderator: Joseph Mihursky
Opening Comments
Joseph Mihursky
I'd like to remind you that support for this workshop
came from the NOAA Coastal Resources Manage-
ment Division through the Maryland Department of
Natural Resources (DNR). This was a combined
effort of Pennsylvania, Maryland, and Virginia. Jerry
Hollowell (Susquehanna River Basin Commission),
Laura Lower (Virginia Council on the Environment),
and Cynthia Stenger (Maryland DNR) were involved
in the planning effort. Dave Pyoas and Steve Jordan
were key people helping in DNR. Close to two
dozen people have contributed their efforts as chair-
men, speakers, conveners, and staff. And of course
the participation of so many people with so little lead
time has been a gratifying response.
Let us consider the long view. An attempt at Bay
restoration is in place. We have acknowledged
problems and need to try to turn off the faucets, and
explore pretreatment. Some information is needed on
priorities, and we need to monitor what's there over
time to discover what gives us the problems. Man-
agement or regulatory initiatives must be tracked in
biological systems. There are already programs in
place for water quality, phytoplankton, microzoo-
plankton, mesoplankton (not all species), and benthic
organisms (quantitative, population). A pilot effort is
also in place to assess fishery stocks using conven-
tional as well as acoustic techniques. This is the
background against which we will begin to deal with
toxics. It's not necessarily a regulatory, litigative
situation. Over time you want to know if you see a
system response in terms of stressors, species diversi-
ty, etc., resulting from a regulatory track. This work-
shop is a first step toward a framework of recom-
mended best approaches.
A document from the workshop will be available to
agencies. The workplan that will utilize the workshop
document will be sent to the Monitoring Committee
for input. Subsequent RFP's will reflect the funding
levels set by September. The funding levels will
determine the temporal and spatial coverage achiev-
able. That is out of our hands. But we can offer our
best science.
Risk Assessment Workgroup Report
Ian Hartwell
Our group discussed a variety of factors influencing
how you do risk assessment in a diverse, complex
system or in a pilot study, and how to formulate
hypotheses. We asked whether and how we can
apply risk assessment techniques to studies like this to
answer the questions being asked. As a result, we
developed an initial ranking scheme as recommenda-
tions to other groups, including ideas for how to
combine the recommendations in ways that are valid
to address questions. Analyzing the relationships to
various factors responsive to the tests is recommend-
ed. The framework is done; details are lacking. Our
ranking scheme is designed to help future efforts -
long-term monitoring plans. We still need to identify
areas needing work.
A ranking system of five parameters was devised.
The parameters are:
Consistency of results
Severity of endpoint
Degree of response
53
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Number of tests
Reproducibility of results
Consistency refers to the agreement between the
different bioassays the other workgroups are devising.
For example, for a given test endpoint, what propor-
tion of the species tested showed a positive result.
For a given species, what proportion of the tests show
a measurable effect. If the results from all tests
and/or species agree, consistency is high, and confi-
dence in predicting toxic effect (or lack of effect) is
high. If only half the test results are positive, consis-
tency is low. Thus the lower internal value for
consistency is 50%.
Severity refers to the degree of effect which the
bioassay endpoints measure. Mortality is the most se-
vere response. Impaired reproduction is second, and
impaired growth is third. Other endpoints can be in-
cluded in the lists.
Degree of response is a measure of the proportion of
organisms responding in each test. Unlike traditional
methods of reporting toxicity bioassay results (e.g.,
LC50), risk assessment and hazard assessment utilize
all response data. Thus a response level of only 10%
is as important to know as the 50% level.
Both severity and degree of response are straightfor-
ward parameters if only one type of bioassay is run at
each site. However, we anticipate that the other
workgroups will propose a suite of tests to be run at
different physical-chemical environments throughout
the Bay system. Thus some system of factoring in
the relative sensitivity of different tests and different
tests species will be necessary to arrive at a ranking
value for these two parameters after the other work-
groups have made their recommendations.
The number of tests run at each site should be the
same for statistical and experimental reasons. How-
ever, given the uncertainties of field work, this may
not always be possible. The suggestion was made
that if some sites don't have the same number of
tests, the equivalent tests at other site should be
discarded. Consensus could not be reached on the
point but most participants did not recommend
throwing out good data.
Rank of these criteria. (The group voted to weight
the factors: l=low, 10=high.)
Consistency
Severity
5.8
8.5
Degree of response 7.9
Number of tests 4.8
Reproducibility 6.6
Other discussions of the group evolved into the
following list of further recommendations:
Define hypothesis specifically. What do you
hope to learn?
Refine experimental design. Locate where you
know you can show problems, and use one such
spot as a test site to demonstrate that your
methods work and to test the ranking system to
see if it works.
Chemical cause-effect studies. What sorts of
chemical contamination have which effects, i.e.,
which effects are most important to look at it?
Obtain an analytical hierarchy process. There is
a software program for this, and consensus is
that it's excellent for this use.
Sample sediment and water together ~ in time
and space.
54
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Include temporal and spatial variation. Multiple
samples and few sites is better than many sites
and few samples.
Endpoints should address population parameters --
i.e., subtler effects.
Q: Are there no ecological risk assessment
models for terrestrial ecosystems either?
A: The only exception I can think of is
models of effects of gaseous pollutants
on forests.
Do water and sediment analyses at the same
point. To predict in a probabilistic sense you
have to have data on the species.
Develop modeling techniques to link bioassay and
population impacts.
Create a database on relative sensitivity of assay
species and endpoints. This recommendation
bears on the debates about consistency, severity,
and degree of response.
Fill out internal items of ranking factors after
assay methods are chosen.
Questions
This workgroup's primary job would
seem to have concerned carrying out the
recommendation to develop modeling
techniques. Can you give any specific
suggestions for such techniques?
A variety of modeling approaches are
available to link bioassays with pollution
impacts. They are not universally appli-
cable or agreed upon. This is a generic
recommendation - there was not enough
time at the workshop to come up with a
tight model linking these things. This
will have to be a long-term effort. As
you will remember, we don't yet have
the dose-response data that the risk as-
sessment models usually use. This prob-
lem hasn't been approached by risk mod-
elers yet.
Q: Could you substitute percent-dilution data
for dose-response and apply this method
to aquatic systems?
A: Yes, except we aren't going to have
dilution data.
Whole Organism Workgroup Report
Steve Schimmel
Our group tried to answer the questions in the work-
group charge (see questions at beginning of this
workgroup's Discussion Notes in Appendix A). We
considered the first question the primary one. To
answer it, we developed a table of methods - species,
duration, and type of test to be used. We were
conservative in our approach, basing our draft recom-
mendations on tests already established and accepted,
such as NPDES permits and pesticide registration.
Methodologies that have not proved workable were
not included.
You will also notice from the chart that second-level
tests are not as precise, but include species important
to the Bay. Third-level tests basically remain in the
research realm, but may in the future be of use.
By answering question 1, we also essentially an-
swered numbers 2-5.
We gave some attention to suborganismal measures
(question 6). Our consensus position was that if we
can reasonably incorporate these measures, we should.
The major question is on costs. Conceptually and
philosophically we think suborganismal tests should
be included. The managerial reaction is questionable.
55
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But the future of aquatic toxicology will probably rest
with these suborganismal tests. We recognized that
some biomarkers are best used not with toxicity
testing but with field collection of indigenous organ-
isms, in tandem with pathology. The estimated
causes and effects once you see toxicity are where
these can be pertinent.
For the next question (7), we did not work much on
the analytical chemistry associated with these tests.
Toxicity data should drive analysis as much as
possible. That is the economic reality. First you find
an effect, then you look to analysis to explain it.
As far as the risk assessment discussion, we reached
no new conclusions. The top tier of tests gives
population responses that might be needed. There is
not much that gives "r," or time to reproduction -
only Ceriodaphnia.
The tests discussed here apply to laboratory situations
~ either static or mobile. We did discuss in situ
testing, but there's not enough data to be able to
discuss it except as research.
The whole organism group agreed with the sediment
group that there should be a control Chesapeake Bay
site. Questions about controls are not resolved.
Questions
Comment: The striped bass larval test is useful only
in the lower end of the salinity range of the estuarine
cell ~ they can't survive in 25 ppt water.
Comment: Biomarkers people would adopt an addi-
tional strategy. Adding biomarkers into acute and
chronic testing makes sense. At the same time, there
is a need to incorporate this approach into traditional
environmental living resources monitoring. We can
gain a lot of information by looking at indigenous
organisms, either at the biochemical or biological
level. I will try to draft a document on how this fits
into our overall goals, circulate it, and aim for pro-
ducing a position statement.
Comment: Maryland DNR thought about this some
years ago. They did a bit of it, but it was terminated
for lack of funding.
Whole Organism Tests for Ambient Toxicity In Freshwater, Estuarine, and Marine Environments of the Chesapeake Bay
Category
Proven Regulatory
Methods
Established Methods
Pertinent to
Chesapeake Bay
Research Methods
Pertinent to
Chesapeake Bay
Freshwater environment
C0riodap/mta 7-day chronic
Fathead minnow 7-day chronic and
embryo/larval test
Stlenastrum 96-hr
Embryo larval: Muegin, catfish
Striped bass larval toxicity test
Duckweed 96-hr
Estuarine environment (max. 25 ppt)
Sheepshead minnow 7-day chronic and
embryo/larval test
Mentta b«y*na 7-day chronic
MysWopste bahia 7-day chronic
Bivalve larvae:
Cr*ssostr»a virginica
Uyaannaiia
StoMonamt algal test 48 hr?
Grass shrimp larval acute lethaity, 96 hr
Striped bass larval toddty test (tow sairity)
Anchoa mitchm 96-hr test
CaftagtasM algal chronic
Sago pondweed toxicity test
Eurytomx* aflWs
Naomysis a/mrfcan* 96-hr acute
/fcarfatorm
Marine environment
Sea urchin tertiization test
Meridia beryllina 7 -day chronic
Sheepshead minnow 7-day chronic
Mysidopsis bahia 7-day chronic
Bivalve larvae test' Mytilus edulis
Champia parvula reproductive
Grass shrimp larval acute lethaity. 96 hr
Striped bass juvenile 96-hr acute lethaity
Atartiatonsa 7-9 days
Anchoa mflchid 96-hr acute lethality (larval)
56
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Sediments Workgroup Report
Richard Peddicord
Sediment toxicity tests can be very useful for some
purposes. Major points from our discussions are sum-
marized below.
Sediment quality criteria ~ Even though they are
still under development, we should collect the pa-
rameters necessary to calculate compliance,
insofar as we know what the parameters are.
There is a powerful consensus that we need to
pull together all relevant information on toxics in
Chesapeake Bay before we design field opera-
tions. Then we can determine what's appropriate
to do, where, whether emphasis should be on the
water column or sediments. All this should
precede design of field sampling.
Most of the off-the-shelf sediment toxicity tests
are adaptations from water tests. We have a need
for tests aimed specifically at sediments and
population endpoints. Short-term mortality of
adults or juveniles is the most common endpoint
among presently available sediment tests. We
need to see more tests addressing the next genera-
tion.
Sampling for toxicity testing should include some
sites expected to give very strong response. We
can work across a gradient to see how well toxici-
ty tests correspond to our conception of what's
there, and see what kind of sensitivity is present.
Does a zero result indicate low contamination, low
bioavailability, or low sensitivity?
In surveying the Bay for degree of degradation, it
is important to look not just at sediment toxicity,
but also at benthic community information and
sediment chemistry information.
There are two types of exposure methods: those
using whole sediments and those using extracts of
sediments. Tests using pore water and organic
extracts of sediments are viewed skeptically. If
we apply tests in the near-term, we must use tests
and species with well-established databases. We
need to anticipate issues of lab maintenance or test
organisms and variability in response.
We need to think in terms of a suite of different kinds
of toxicity tests ~ different species, different tests.
As a rule of thumb, amphipods are among the more
sensitive species, and we have concentrated on them.
However, we still need other species in which there
is an indication of relative sensitivity.
In developing tests, we should emphasize some
indigenous Chesapeake Bay species, with at least
some endpoints that address the next generation's
success.
It is extremely important to develop protocols related
to the specific objectives of this program, standard
operating procedures, and quality control/quality
assurance measures before getting very far into the
studies.
Studies of bioaccumulation could consume great
quantities of time and money, and should not be part
of an initial survey. Where problems seem likely, we
could look at potential linkages between sediment
and tissue contaminants. This would provide a
context for interpretation. These techniques should be
used in the framework of a hypothesized problem
(such as contamination in fish for human consump-
tion), not in a survey situation.
Work on biomarkers should be encouraged, but they
are not very useful at present. They will become
more useful when links have been developed between
biomarkers and whole organism responses.
57
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The final point in risk assessment is relevant to the
entire effort in ambient toxitity assessment: At the
program level and below, we need carefully defined
objectives in the form of testable hypotheses. We
must clearly state exactly what we are trying to
determine.
Questions
Q: Has any attention been given to resus-
pended sediments as opposed to those in
place?
A: No - our group didn't work thai, broadly.
Resuspension of sediments is important
in terms of contaminant bioavailability.
Q: Was fluidized mud addressed?
A: No.
A closing observation concerns appropriate controls.
We talked about controls and reference sediments.
The relative importance of that question may be
dependent on the objectives. If you're doing a survey
of intensity of response, it may not be very important.
For hypothesis testing, it is crucial. Thus selection of
appropriate reference points involves managerial as
well as technical issues: do you want a comparison
with pre-John Smith sediments? or with the cleanest
you can find now?
58
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BIOASSAYS FOR SEDIMENTTOXICITYTESTING IN FRESHWATER, BRACKISH SALTWATER, AND
HIGH-SALINITY ENVIRONMENTS IN THE CHESAPEAKE BAY
Organism
1
2
3
4
5
6
7
8
9
10
11
12
Notes
Standard bioassays
Freshwater
Hvallela
Chironomus
Hexagenia
Brackish saltwater (0-1 5 ppt)
Hyallela
Eohaustorius
High salinity
Rhepoxinius
Amoelisca
Bivalve larvae 48-hr
pediveliger oyster
Mercenaria
Polychaete Neanthes (Nereis}^
Mysidjgrass shrimp)
X
X
X
X
X
X
X
X
X
?
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
So
So
So
So
So
So
ss
So
So
a,b
a,b
a,c
a,d
L
L
L
L
L
L
L
L
M
M
Bioassays available based on
enhancement of standard
. techniques
Low salinity
Leptocheirus
Eohaustorius
High salinity
Lepidactylus
X
X
X
X
?
?
?
X
X
X
X
X
X
X
X
Bioassay techniques under develop-
ment or recommended for
development
Chronic tests
Leoiocheirus
Lepidactylus
Sago pondweed
X
X
X
X
X
X
X
So
So
So
M
M
M
X
So
So
$0
H
H
H
(10) So-»<*d ptMM; 80-««dlm«nt •Jorry
(11) cxacute Mhafc b«b»h«vter«l •ftoct; c-aboorm«l d«v«lopm«*; d>4iMtMnorphic Wkira, «te.
(12) L-tow cott; M>modwate coat; H*high co«t
59
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General Perspectives on the Role of Bio-
markers (Biochemical Measures of Ef-
fects) in the Chesapeake Bay Toxics
Workplan
(Comments compiled from Dr. Ken Jenkins, Dr. Brian
Bradley, Dr. Gun Roesijadi, Dr. Margaret James, Dr.
John Pritchard, Dr. Wolfgang Vogelbein, and Dr.
Dave Wright, and submitted by Dr. Jay W. Gooch,
suborganismal plenary session speaker and convener)
Although the suborganismal responses workgroup
session was not formally convened at this workshop,
a number of investigators with research experience in
this field were present. This document has been
prepared in collaboration with those present and is
intended to reflect other important comments and/or
suggestions related to the use of biochemical effects
measurements for the assessment of ambient toxicity
in Chesapeake Bay waters.
The general consensus of the workgroups was that
biomarkers were clearly the tools of the future, but
concern was expressed that they may not yet be in a
form where they can be used on a routine basis. We
propose, however, that subcellular biomarkers (bio-
chemical effects measurements) can be used effective-
ly in conjunction with the conventional bioassay
studies that were the major topic of the workshop.
Carrying out a range of biomarker tests in conjunction
with the proposed bioassay program would provide a
number of important advantages to the Chesapeake
Bay Toxics Program:
Subcellular biomarkers are substantially more
sensitive to toxins than conventional bioassay end-
points, and their implementation in this program
would provide a more accurate picture of the
distributions of low-level toxins in the Bay.
Unlike conventional bioassay endpoints, subcellu-
lar biomarkers can provide information on the
types of pollutants responsible for any observed
toxicity.
Carrying out subcellular biomarker tests in con-
junction with well-characterized whole-organism
bioassays will allow these new methods to be
calibrated and validated in a well-defined and
experimental framework. These tests will provide
a basis for rigorously defining the
relationship between subcellular endpoints and
parameters such as growth and reproduction.
This provides a cost-effective approach to opti-
mize the information obtained from the toxics
program.
We propose that subcellular biomarkers can also be
used effectively with ongoing field survey programs.
In these studies, biomarker assays can be performed
on native organisms and the results compared with
both chemical and biological data, which are normal
components of these programs. We recommend that
indicators of contaminant-induced changes be mea-
sured in conjunction with ongoing sampling programs
being conducted in the Bay. The numerous sampling
efforts aimed at the monitoring or evaluation of
populations of Bay species could accommodate these
studies at more modest cost than initiation of a new
program. As significant resources are already devoted
to the task of sampling resident species, it appears
economically wise to extract as much information as
possible from the effort. Again, this approach would
provide information on sensitive sublethal effects in
native organisms and provide insights into the caus-
ative agents when toxicity is observed. It would also
allow biomarker endpoints to be correlated with both
chemical and biological field data to further calibrate
and validate these procedures.
In his plenary presentation, Mr. Steve Schimmel of
the EPA's Narragansett Laboratory suggested that
60
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acute toxicity bioassays conducted on ambient waters
of the Bay would almost certainly be negative, as has
been the case for most of the ambient toxicity surveys
conducted by the EPA. That is, experience suggests
that acute toxicity problems remote from known
sources of pollutants are very rare. In fact, chronic
effects, as defined by bioassay protocols, are also
somewhat rare. Despite this, many coastal waters like
those of the Chesapeake Bay are experiencing popula-
tion declines of ecologically or recreationally impor-
tant species. It is widely perceived that at least some
of this decline may be due to pollutants. Biomarkers
may provide the mechanism to detect consistent,
repeatable measures of chemical stress in indigenous
biota.
Bay Programs which may be able to incorporate
pollution studies:
• State finfish and shellfish contaminant monitoring
programs
• Benthic infaunal surveys
• Fish and shellfish population surveys (stock
assessment)
Habitat surveys
• Field-oriented research programs (Sea Grant
projects, etc.)
Used in the appropriate context and with the appropri-
ate questions in mind, incorporation of biochemical
effects measurements into ongoing sampling programs
in the Bay should provide important additional
information regarding the effects of toxic chemical
contaminants.
References
Vernberg, W.B., A. Calabrese, P.P. Thurberg and F.J.
Vernberg (eds.) 1987. Pollution physiology of estuar-
ine organisms. University of South Carolina Press,
Columbia, 458 p.
Versteeg, D.J., R.L. Graney and J.P. Giesy 1988.
Field utilization of clinical measures for the assess-
ment of xenobiotic stress in aquatic organisms. In
W.J. Adams, G.A. Chapman and W.G. Landis, eds.,
Aquatic Toxicology and Hazard Assessment: 10th
Volume, ASTM STP 971. American Society for
Testing and Materials, Philadelphia, pp 289-306.
White, H.H. (ed.) 1984. Concepts in marine pollution
measurements. Maryland Sea Grant Publication
number UM-SG-TS-84-03. 743 p.
61
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Appendix A
Workgroup Questions and Discussion Notes
The workgroup sessions were instructed to address specific questions on the topics. These questions are attached
to each workgroup's discussion notes. The highlights and points of consensus for each workgroup were synthesized
and presented in the final plenary workgroup reports.
Questions Related to Population Risk Assessment
The objective of the Risk Assessment Workgroup is
to develop a risk assessment scheme (preferably
quantitative) which can be applied to the whole
Chesapeake Bay or its subsystems, for the purpose of
risk management at the state and
federal government level. Secondary uses of the
scheme are to guide monitoring strategies and provide
damage assessments. In arriving at such a scheme,
the following questions and subsidiary considerations
should be addressed.
I. Technical questions
A. What is the mathematical approach?
1. qualitative
2. quantitative
3. other
B. How to compartmentalize the model to
accommodate input?
1. whole organism toxicity
2. sub-organismal toxi-
cology
3. sediment toxicity
4. physiographic regions
5. specific sites
D.
How to estimate chemical exposure?
1. water
2. sediment
3. atmospheric
4. discharges
5. fate and transport
6. other
How to define receptors and endpoints?
1. field biomonitoring
2. lab studies
3. surrogate/indicator spe-
cies
acute toxicity
sublethal responses
population/community
responses
other
4.
5.
6.
7.
E.
F.
G.
H.
How to incorporate temporal variations?
How to calculate uncertainty?
How to field verify?
What are the model outputs?
1. risk predictions
2. monitoring guidance
3. research guidance
63
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II. Practical questions
What do the states want?
A. Has the approach been used elsewhere
(does it work)?
B. What are the operational requirements?
1. level of personnel
expertise
2. time commitments
3. computer facilities
4. cost
C. What are the data requirements?
1. what kind of data
2. how much data
Workgroup Sessions on Population Risk
Assessment
Convener and Chair: Ian Hartwell
Most of the research of the Chesapeake Bay Program
has been focused on nutrients, not toxics. The 1987
Chesapeake Bay Agreement has various objectives for
Bay restoration, including a Toxics Reduction Strate-
gy, among other items. Our objective at this meeting
is to provide scientific comments on the strategy and
the pilot biomonitoring study.
One of the goals of the strategy is to produce quanti-
tative risk assessment on commercially, recreationally,
and ecologically important species, and on the system
itself.
The Basinwide Toxics Reduction Strategy states that
information generated from the pilot phase assessment
will be used to estimate a risk. No assumptions will
be made about chemical exposure; rather, biological
effects will be used to assess the need for further
study.
The workgroup did not agree with this view. Risk
assessments cannot be created in the absence of
chemical exposure information.
Steve Jordan (Maryland Department of Natural
Resources): The Bay Agreement of 1987 has as its
primary goal to protect the Bay's living resources.
We have nutrient goals that we think will help and
other ways of improving water quality. The toxics
area is the weakest. The Basinwide Toxics Reduc-
tion Strategy is more specific. Commitments include
a toxics loading inventory. We want to use bio-
assays to estimate risks to populations of living
resources. The key question is how to extrapolate to
real populations. It is the populations that managers
care about, not individuals. Managers need to know
whether an important population is at some risk, and
they need to know the magnitude of risk and the
confidence level of our prediction.
The Virginia participants agree with Jordan.
Question: Do we want to develop risk assessment or
damage assessment?
These items were discussed as distinct entities. The
first is a predictive assessment while the latter is
retrospective in nature. The effects of toxics as
opposed to confounding parameters such as acid rain,
anoxia, turbidity, overfishing, etc., were discussed.
These items must be taken into consideration when
interpreting the field bioassay results. It was decided,
in light of the proposed workplan, to adopt a tiered
approach.
Comment: We must be careful when demonstrating
cause and effect. The mechanism may not be known,
so we should be cautious in pursuing these mecha-
nisms, being wary of confounding factors.
We must also address techniques that can incorporate
mixtures, since we're really looking at a toxic soup.
That is the purpose of subletbal biological assays.
They allow us to examine whether inputs like acid
64
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rain and pesticide runoff are causing problems in
addition to those caused by effluents.
Identifying these problems should be the first step
addressed in the field. Correlating the issues of cause
and effect is the next problem. Where you have
contaminants you're likely to have high organic
loading. We will need to differentiate the toxic
effects from the effects of organic loading. But first
we will demonstrate that there is a problem, and then
demonstrate the cause/effect relationship. Then the
third step is to look at what is in the mixture so we
can concentrate on that particular chemical or series
of chemicals. This is analogous to the NPDES
permitting process. The permittee analyzes the
effluent and then must figure out what part of the
effluent needs to be cleaned up. They reconstruct the
effluent or fractionate it in order to find the culprit.
We will need to do the same thing.
Point: Those are management questions. We need to
focus on an assessment system that will allow the risk
managers to find the culprit and the organisms at risk.
Risk assessment also needs to address other factors
like anoxia.
Response: We need a tiered system that asks, "Do
we or don't we see problems?" We need a tool to
look at stresses; and if we find them, then look
deeper. The next question is how to manage that.
We have to ask whether it's toxics-related, or an
effect of habitat or another impact like overfishing.
Then we ask, "Is it a pollutant? What type of pollut-
ant is it?" Fractionation doesn't solve the problem
because it may change the overall toxicity.
Comment: We will be working in the context of
information that is already fairly well described.
With a tiered approach, we've done the first tier; we
know that there is a problem and that it's not totally
pollution-related. There are other problems such as
overfishing. To address the toxics, we should focus
on mortality, growth and development, and reproduc-
tion of organisms. These are the critical factors for
maintaining a population. These can be used as
categories, and other factors can be related to them.
Assays should reflect these categories. Otherwise
there will be problems.
Question: Will we collect chemical data?
According to the proposed workplan, sediment and
water chemistry data will not initially be available.
The group strongly urged that at least routine water
quality data (i.e., O^ salinity, pH, turbidity, etc.) be
gathered at each test site. Abo, any biological tests
performed should be selected such that the data can
address or be capable of being extrapolated to
community effects. In the absence of chemical data,
loading estimates at a known contamination she could
be used to exposure estimates in a risk assessment.
The option of creating a risk assessment vs. a damage
assessment was revisited.
Response: That type of data collection will be driven
by the results of the first biological analysis.
Response: The draft workplan suggests that we
begin with field assays and apply chemical analysis
only where we get positive results, and preferably for
implicated chemicals. There is already a lot of body
burden information but it does not tell us anything
about population level responses.
Comment: You should definitely collect general
water quality parameters at the same time, to elimi-
nate other causes such as DO.
Response: These systems are already well character-
ized for this.
Point: We should come up with a rational system to
take field bioassay data and find out if it's telling us
that toxicity is a problem. Risk assessment involves
exposure assessment, which involves field observa-
tions — biomonitoring, etc. Pre-supposing that you
65
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know the toxicity, you have the exposure component
and the toxicity.
Response: Yes, but we don't have input on exposure
assessment, so should we do risk assessment or
damage assessment?
Question: We will have information on growth,
mortality, and reproduction; what will we do with
this?
Response: Damage assessment is taking stock of
what has occurred. It is retrospective, as opposed to
risk assessment, which is predictive. Also, there is
hazard assessment, which is the inherent clanger of
chemicals,
Question: The state wants a method that has been
used and is known to work. Is there such a model or
approach already developed that could use this
information?
Question: Should we be looking at risk to popula-
tions, or are we interested in what will be expected to
happen to the community?
Response: It could be both. For both populations
and communities the information acquired must
encompass the representative trophic levels.
The workgroup discussed the types of data required.
For assessment purposes dose response data or re-
sponse surface calculations are more useful than
LC50 or EC50 data. Also sublethal endpoints are
better than lethality tests.
Point: We should divide the Bay into geographical
areas and then look at the hydrological, biological,
and ecological effects. Also, we must assess the cost
and prioritize what we really want to do.
Question: What can we do with the pilot scale, and
then what do we want from it in order to interpret the
Bay in the long run?
Response: We need to proceed with a qualitative
system, ranking information from pilot studies, and
we need to be able to expand it.
Response: No, I think we will have to get qualitative
and quantitative data depending on the problem we
are addressing. For example, if a chemical is a
carcinogen, we don't even have to subject it to other
tests.
Comment: This won't happen. We will not have
chemical data up front.
Response: But it can, and other tests must appropri-
ately assess hydrological, geological, biological, and
ecological factors. We have to figure out what our
strategy is.
Comment: It is important to let the other workgroups
here know whether we want hypothesis testing or
continuous data.
Point: A comprehensive toxics database for the Bay
does not exist. The primary need of the states is to
determine how to handle monitoring data; that is,
sublethal toxicity data. Are there toxics problems
outside of hot spots? And how can we best answer
those questions? What is the best way to handle the
data?
Response: We need to stop presenting results of
tests in terms of statistical differences. Results may
vary greatly. So we should tell the lexicologists to
express results in terms of magnitude. We need data
more like dose-response; not pair-wise comparison.
How will we infer causation from those kinds of
data? There are potential mistakes, like inferring
toxicity when you're really seeing differences caused
66
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by physical factors. Also, to catch episodic events
their strategy must be designed appropriately.
The workgroup discussed in more detail the impor-
tance of specific endpoints in light of the proposed
workplan limitations and the tiered assessment
approach. The biological tests should be related to
population effects. Field studies are more relevant
but may be difficult to interpret relative to toxics
effects. Toxicological studies are more straightfor-
ward but results must be extrapolated to the field.
Validation studies are required
Question: OK, but what do we tell the state? I
suggest we do it in terms of a tiered approach.
Response: I'll design a proposal to tell the state
where we are, which is essentially a statement of
need. It can provide a workplan with various tasks,
with a number of scenarios, a schedule, and an
estimate of how much it will cost. We should write
a proposal and let them fill it in.
Comment: A lot of that exists. We should decide if
it is good, and if not, modify it. This strategy is a
draft and we do have input. We should decide if the
tasks are adequate or not.
Point: Other groups are proposing that in addition to
the Potomac, we should consider adding Baltimore
Harbor and the Elizabeth River. Also, the Statement
of Purpose calls for monitoring of effects on living
resources. This is to learn effects, not to establish
regulations or anything else.
Question: Take a scenario. If we see a strong
gradient of toxic effects down the Potomac in the
pilot study, how do we use that data to tell the state
what they should do next?
Although this question was not answered, most
participants felt that the study design was flawed.
Some participants feel that we need to define the
endpoints so they are sure to be relevant to biological
toxicity and effects on populations.
Question: Why don't they just do field studies?
Response: That can be very difficult. For example,
Hall did a study of larval striped bass in the Nanti-
coke River, and mortality occurred in river spawning
grounds, but not in the control (Vienna town water).
This indicated that something in the water was the
cause of the problem. This has since been thought to
be low pH and high Al levels.
Point: We should address the package that the state
has given us to find its limitations and a way to
optimize this structure. Then we can tell them what
they can get, and also what they can't get from the
proposed structure.
Question: Let's assume the state has given us
toxicity information for a fish, an invertebrate, and a
phytoplankton species. Now what do we do?
Response: Give the state a system to rank the sites
by degree of response. Produce a qualitative ranking
system.
Comment: It's hard to differentiate the noise level
from the effect of toxics. Theoretically, if mortality
in striped bass from larvae to juvenile is normally
95%, an increase to 96% (which is probably not
detectable) will halve the number of recruits.
Question: So what is the significance of the ranking
system?
Response: What endpoint would we be most inter-
ested in, and what would bother you the most?
Remember that direct toxicity to some organism
implies indirect toxicity to other trophic levels.
67
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Comment: They may have to develop a more sophisti-
cated community level study like the MERL meso-
cosms.
Comment: From the Nanticoke example we can see
that they didn't find the problem. If we knew why,
it would be helpful to us.
Point: You can't rely on toxicity tests alone. You
have to incorporate field studies. Beware of false-
positive results, or false-negative results.
Comment: The SAV decline, thought to be due to
herbicides, was decided to be due to shading, and
now they say it may be herbicides again. In some
areas the most sensitive lifestage --germination - was
not studied due to lack of ability to handle the seeds.
Comment: We need suborganismal tests for this
reason.
There was some discussion of what we learned from
our experience with the SAV decline and how to apply
it to the ranking scheme.
The workgroup discussed how to characterize site-
specific results in a ranking scheme to contrast
different sites. Tests employing more than one
species yield more information but require sensitivity
calibration. Field effects may be due to direct toxicity
or indirect community effects. The group strongly
urged that sampling be done for both sediment and
water at the same place and time, and that the draft
workplan be modified to increase spatial and tempo-
ral replication. It was suggested that sites known to
be contaminated be included in the effort. Also, the
hypothesis to be tested in the monitoring study needs
to be clearly defined and test endpoints should be
selected accordingly.
Question: But what's going to be in the ranking
system? We should provide the states with a ranking
scheme so that they can do site characterizations.
Comment: We have done site characterizations, but
they have been waste sites.
Question: Given a suite of sites, can we develop a
preliminary methodology to determine whether there
is a risk? That is, with some group of test species,
what parameters would be good to look at?
Comment: It will be important to know the severity
of the endpoints and to have consistent results. Also,
we want the severity of effects to be measured in a
graded fashion, as opposed to "yes or no" answers.
And we should also use multiple types of species in
consistent studies.
Question: Should a species be chosen that can
tolerate a wide salinity range?
Point: We must consider the sensitivity of the test
species.
Point: Inter-species variation can be handled statisti-
cally. However, we should decide whether we'll use
the same suite of test species or whether that
can be variable. For ranking, relative sensitivity will
vary with the substances and other factors that are
involved.
Point: There is a problem, though, because it may
affect a parameter like competition which, in turn,
may affect the endpoint.
Response: That's all right since the endpoint is still
being affected by the toxicant, even though the effect
may be indirect.
Point: They should do parallel testing of the sedi-
ments as well as the water.
Point: Many variations such as temporal variation
need to be considered.
68
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Point: You may want to add less severe endpoints so
you can see low impact areas as well.
Question: But what if negative effects are a result of
our ignorance, not of the lack of toxicity?
Response: We should have a strategy to choose
stations.
Response: They do have a strategy as explained in
the document.
Point: If they expect to draw a relationship between
the effects and toxicants as a cause, they should
choose sites where they have the maximum likelihood
of that occurring.
Point: Don't overemphasize the number of locations
tested at the expense of replication and observation
over time. There may be variation due to time of
year, weather, species present, and the like.
Point: Keep in mind that in an estuary, it's hard to
establish a pollutant gradient system because you're
moving in and out of many physico-chemical sub-
gradients, which have different impacts on pollutant
effects.
Point: All we can recommend is a meaningful way
to form the sampling strategy, because we don't have
a budget.
Point: Field tests should be carried out in an area
where you know effects exist.
Comment: Information exists that the tests do work.
It's the temporal and spatial effects and the like that
would cause variability.
Comment: A previously-derived report may serve as
an example here. This was an NRC panel specifically
addressed to monitor the Chesapeake Bay, to deter-
mine how we could design a monitoring program.
They had similar concerns on temporal and spatial
processes.
Question: Are these types of sampling sites appro-
priate? necessary?
Response: If the hypothesis is that there is no
measurable toxicity in the ambient waters or sedi-
ments in the Chesapeake Bay, you still need to get
into severity and degree.
Point: We need water and sediment chemistry, but
that can't be done right now.
A ranking system of five parameters was devised.
The parameters are:
• Consistency of results
• Severity ofendpoint
• Degree of response
• Number of tests
• Reproducibility of results
Consistency refers to the agreement between the
different bioassays the other workgroups are devising.
For example, for a given test endpoint, what propor-
tion of the species tested showed a positive result.
For a given species, what proportion of the tests show
a measurable effect. If the results from all tests
and/or species agree, consistency is high, and confi-
dence in predicting toxic effect (or lack of effect) is
high. If only half the test results are positive, consis-
tency is low. Thus the lowest internal value for
consistency is 50%.
Severity refers to the degree of effect which the
bioaassay endpoints measure. Mortality is the most
severe response. Impaired reporduction is second, and
impaired growth is third. Other endpoints can be in-
cluded in the lists.
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Degree of response is a measure of the proportion of
organisms responding in each test. Unlike traditional
methods of reporting toxicity bioassay results
(e.g., LCM), risk assessment and hazard assessment
utilize all response data. Thus a response level of
only 10% is as important to know as the 50% level.
Both severity and degree of response are straightfor-
ward parameters if only one type of bioassay is run
at each site. However, we anticipate that the other
workgroups will propose a suite of tests to be run at
different physical-chemical environments throughout
the Bay system. Thus some system of factoring in the
relative sensitivity of different tests and different tests
species will be necessary to arrive at a ranking value
for these two parameters after the other workgroups
have made their recommendations.
The number of tests run at each site should be the
same for statistical and experimental reasons.
However, given the uncertainties of field work, this
may not always be possible. The suggestion was
made that if some sites don't have the same number
of tests, the equivalent tests at other site should be
discarded. Consensus could not be reached on the
point but most participants did not recommend
throwing out good data.
Reproducibility is the statistical measure of variability
within a tests at a given site.
Candidate System
Comment: There is a software package by Thomas
Saaty at Pitt. The name, AHP, stands for Analytical
Hierarchy Process. It is not predictive, il simply
takes qualitative evaluations and develops a ranking
system. It can compare many factors and is relatively
inexpensive.
Many of the committee members are familiar with
this, and there seems to be general agreement that
such a package would be helpful.
Question: Where are you measuring the uncertainty
of effects?
Response: Under "degree of response."
Point: Anything that we come up with is also useful
in other locations outside the Bay where similar envi-
ronmental situations exist. This is important because
of the lack of actual estuarine bioassays.
Question: Areas are chosen largely due to their
importance to commercially important species.
Should this be of concern?
Question: How do we utilize the endpoints from the
other workgroups? Is transferability among sites and
organisms important?
Response: If the information transfers to other areas,
good. But that is not our task. However, if it doesn't
transfer, we have to ask whether it is because the
system we are working in is unique (due to size for
example), or because the scheme we have proposed is
not good. Transferability is the sign of a good
scheme or theory.
Point: We should use a Chesapeake Bay organism.
But we should be able to transfer to different systems.
Can we address specificity to a given chemical?
There is agreement that the protocol will not address
specific chemicals, or even types of stressors, i.e.,
anoxia and nutrients.
Comment: We should assign weight to each factor.
Response: No, just use analytical hierarchy process,
AHP. A group of experts should determine the
ranking scheme for the AHP. This should be a
suggestion of this committee.
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Which of the ranking criteria are most important?
As an initial evaluation of the ranking factors, each
member of the workgroup assigned a score to each
factor to reflect its relative importance.
Ranks were tallied on a scale of 1-10, and results
were:
Consistency 5.8
Severity 8.5
Degree of response 7.9
Number of tests 4.8
Reproducibility 6.6
These values could be used as weighting factors, but
would require validation to use. The values here
represent only the quick polling of a small group.
Questions to workgroup
I. Technical questions:
A. The Mathematical approach will be qualitative;
however, input data is quantitative. The approach
may become quantitative as the monitoring process
develops.
Comment: You will have chemical data if you get a
positive from the preliminary screening. We should
do chemistry if the biology dictates it. If sample
fractionation is used, you do have chemical informa-
tion.
Comment: You should also do chemical analysis on
samples that are not positive to see what is in the
samples that are not toxic, and compare these to the
toxic samples.
Comment: The chemistry should be done to validate
the endpoints and species chosen, but it will be hard,
due to cost.
Comment: Include chemicals that you think may be
a problem. For example, look at the pesticide di-
mib'n. We could find areas where it has not been
sprayed to use as reference sites.
B. Compartmentalization of both sediment and whole
organism toxicity will be measured by sublethal
parameters, and if appropriate, so will any proposed
suborganismal tests.
C. Chemical exposure estimates cannot be done
directly as planned without chemical sampling. It
could potentially be done later or with historical data
if the site is known.
D. Receptors and endpoints can be defined after
looking at field biomonitoring, sublethal responses,
and surrogate indicator species. This will lead to the
generation of population/community responses.
E. Temporal variations: The experimental design
should address a gradient and a time factor. If you're
not doing a time-course study, at least note the time
of sampling for future reference.
F. Uncertainty is associated with spatial and temporal
scales, and with sensitivity of species and test sys-
tems. The effective question is how to estimate
probability. Information theory can be used in this
context. Possibilities are the Analysis of Extrapola-
tion Error approach and the Cluster Hypothesis.
G. Field verification: Use the ranking system to
determine a set of sites to go back to and then see
whether this area is hazardous by using a complete
battery of tests. This will verify the method.
H. Model outputs: We don't provide a model; it's
more like a system. It will guide monitoring and
research. If the chosen site has a gradient, then the
basis for choosing the gradient will be historical data.
You can take samples and freeze them, but this may
change the partitioning of chemicals between particu-
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lates and water. You could definitely freeze assayed
organisms for later suborganismal studies.
n. Practical questions
A. Prior use. Ranking approaches have been used
and are accepted, but this particular set of factors has
not been used, since it is specific to this monitoring
program.
B. Operational requirements. The modeling links
may require specialized people, and creating databases
may take a lot of time. We may also need trained
people for this. The group generally agrees that two
or more person-years would be needed.
C. Data requirements. Data should be population-
related. We need practical data, the more the better.
Question: Is there a firm toxics database? It seems
that there may be some data, but it may be concen-
trated in hot spots such as Hart-Miller Island, Balti-
more Harbor, and Elizabeth River.
Comment: PAH contamination comes from combus-
tion products in the atmosphere.
Recommendations for the pilot study
.• Define the hypothesis for the pilot and long-
term monitoring study.
• The experimental design needs a great deal of
refinement with particular emphasis on quantity
and quality of test sensitivity.
• Do specific cause-and-effect chemical studies.
• Look into the Analytical Hierarchy Process as a
ranking system integrator.
• Fill out internal items in the ranking factors, etc.,
after methods are chosen.
• Do sediment and water column tests at the same
time and place.
• Coordinate sampling to consider temporal and
spatial variation.
• Address population parameters using endpoints
from biomonitoring tests.
• Do water and sediment chemical analysis at some
point.
• Develop modeling to link field toxicity bioassays
to population impacts.
• Create databases of the relative sensitivities of
assay species.
Questions Related to Methodologies for Whole
Organism Toxicity Testing
The whole organism toxicity testing session will have
two plenary speakers. Each speaker will separately
address "state of the art" techniques for either labora-
tory or field toxicity tests. The following questions
should be addressed by the workgroup after these
presentations.
1. What are the most appropriate laboratory and field
toxicity tests for evaluating ambient toxicity in the
water column of the Chesapeake Bay watershed?
2. Which laboratory and field tests are appropriate
for the various types of habitats that need to be
evaluated (i.e., streams, rivers, open Bay)?
3. What are the relative costs for each type of test?
4. What are the most appropriate test organisms that
should be used for these tests?
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5. What biological end-points should be used?
6. How can sub-organismal measures be incorporated
into traditional acute or chronic toxicity tests?
How can results be interpreted in terms of [risk
to/impact on] living resources and their habitats?
7. What type of water quality and contaminants data
should be collected during ambient laboratory and
field toxicity tests?
8. Is additional research needed to develop laboratory
and field methods for assessing ambient toxicity in
the Chesapeake Bay watershed?
9. How can data generated from laboratory and field
toxicity tests be used in "risk assessment"?
Workgroup Sessions on Whole Organism
Methodologies
Convener: Lenwood Hall
Chair: Steve Schimmel
Hall: Workgroup participants should refer to the
questions distributed previously and use them as an
outline. The Maryland Department of Natural Re-
sources is planning a 1-2 year pilot study of ambient
toxicity, probably on the Potomac River, to determine
the extent of ambient toxicity problems within the
Chesapeake Bay basin. The results of this workshop
are to be applied to that study.
Schimmel (US EPA): National methodologies exist
for coastal environments and have been applied from
Maine down to Florida and around the coast to Texas,
with separate methodologies for the West Coast. My
presentation yesterday described effluent and ambient
tests representative of the Narragansett, Rhode Island
Lab. These may or may not have direct pertinence
for the Chesapeake Bay system. Perhaps the end-
points are pertinent, even if the organisms used are
not, but our purpose here is to develop a list of
methodologies for the Chesapeake Bay. I am not
partial to any one group of tests; we are working
together to assemble a suite of tests that may be used
here.
Purpose of the pilot project: to develop a suite of
tests that are feasible, pertinent, and useful.
Locations of sites. Maryland is leaning toward using
the Potomac River because it encompasses a variety
of problems encountered in the Bay, including heavily
polluted sources and freshwater areas. It was felt that
exclusion of Baltimore Harbor and Elizabeth River
was unwise, because if ambient toxicity testing will
work at all, it will work at these sites.
Review of points 1-5 in the charge to the workplan,
p.4:
1. biological responses to be measured
2. technology available
3. cost
4. sensitivity to sublethal toxicity
5. significant effects.
In considering the selection of freshwater/saltwater
species for laboratory tests used in the pilot study, the
lab tests should encompass: species pertinent for
regulatory purposes, species not pertinent for regulato-
ry purposes, and any additional alternatives suggested
by the workgroup here.
A good starting point will be the NPDES permit
experiences of Virginia and Maryland. What are the
species they use and what are their appraisals?
Maryland NPDES species:
Acute
Freshwater: Daphnia
Fathead minnow
Brackish: Sheepshead
Silversides
Grass shrimp
Mysid shrimp
Neomysis has also been used.
Chronic
Ceriodaphnia
Fathead minnow
Sheepshead
Mysid shrimp
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Virginia NPDES species:
Acute
Freshwater: Daphnia pulex
Fathead minnow
Brackish: Sheepshead
Mysid shrimp
Chronic
Ceriodaphnia
Fathead minnow
Sheepshead
Mysid shrimp
Champia
Have Fundulus and grass shrimp also been used?
Other considerations. We should also use ecological
indicator (keystone) species, as well as species
amenable to laboratory testing. There is some over-
lap: some of these amenable species are also ecologi-
cally significant species (for example, grass shrimp,
Neomysis, Mysidopsis). Some good test species are
only available on a seasonal basis, for example striped
bass larvae, which is an important species and sensi-
tive stage. In using lab species, we must maintain
"ferality" to assure that we're not testing genetically
different organisms from feral species. We should
also use some species that are common to both lab
and effluent testing. Data comparing the sensitivity
of the silversides/sheepshead minnow to some of the
indigenous species of the Bay are incomplete, but
mysids appear to be preferable for chronic testing.
For an acute test for finfish, Menidia and striped bass
should be considered.
methodologies will be designed to meet the goals of
the Toxics Reduction Strategy. In our considerations,
we will combine elements of both more-standardized
tests in the regulatory sense and broader-based tests
and methodologies. We will address the general
water quality considerations of Chesapeake Bay,
recognizing that specific sites indicating toxic point
source problems will be addressed directly by other
regulatory testing tools (e.g. NPDES).
Point: Acute lethality tests for Bay species are not
sensitive indicators of ambient Bay toxicity and
should not be used routinely as such. Chronic tests or
partial chronic tests with sublethal indicators are
preferred. In areas where you expect high toxicity,
you may want to use acute lethality testing as an
initial screen.
Agreement was made to establish three categories of
tests:
1.
Established regulatory methods
2. Methods for less commonly used j
nous species
3. Methods for indigenous species, still
within research mode.
One could argue for conducting chemical testing
when no ambient toxicity is shown, but to keep living
species as good indicators of toxicity.
Question: What comprises the list of chemicals
present in the Bay, and do these species reflect levels
of these chemicals?
Response: The EPA's National Water Quality
Database would be the source for that information.
Purposes intended for these tests. The suite of
organisms will be vastly different depending on
whether the study has regulatory endpoints. Our test
We also have three environments to consider:
1. Freshwater (0 ppt salinity)
2. Estuarine ( > 0 to 20 ppt)
3. Marine (> 20 ppt)
We need to have (1) benchmark species and (2)
controls/references of water with matched salinity,
both of which can run through testing procedures.
Choice of control sites. The lower Bay on Mary-
land's Eastern Shore is cleaner (based on demogra-
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phy); thus it is a candidate for control sites/reference
area. Discussion on use of reconstituted waters
included the caution that you
may add back higher levels of metals from artificial
sea salts than the levels you would be testing for.
Further options were suggested for sources of control
water.
• Use water from Punch Island Creek, a relatively
unspoiled refuge on lower Eastern Shore, with
high oyster seed population.
• Filter Bay water through a well site to be estab-
lished on a beach.
• Consider multiple sites of varying salinities,
especially critical habitat requirement areas.
* Filter ocean water. Caution: diluting natural
seawater with distilled water gives ionic variance
that is not representative of ambient low-salinity
samples.
• Control for each site or range of area? In this
case there must be general guidelines.
Question: Where will the tests be conducted?
Response: Tests will be done on a multi-institutional
basis and in mobile labs.
Slide: Bay water flow observations made at one point
showed six circulation patterns, with varying percent-
ages of frequency of occurrence. Water samples over
ten days yield an average of these patterns, which is
the dominant circulation. This variability should be
considered during ambient water toxicity testing.
Response: We try to get at this integrative function
for water patterns by looking at the animals — that's
why we do in situ testing.
Point: The longevity of a monitoring program is
inversely related to the individual cost per sample;
high costs ensure short-lived programs.
Response: In initial studies it may be necessary to
oversample so that your long-term program does not
overlook significant factors. It is important to make
the best use of this pilot study.
Costs of composite samples may be worrisome for
ambient testing procedures. There are basic questions
on how to collect these samples. Two options are
seen: grab sampling at various depths, and composite
sampling. For both methods, both cost and design
must be considered. EPA's Technical Support
Document outlines the pro's and con's of these
sampling methods.
Stationary vs. mobile laboratories. It was agreed
that there are inherent differences in the use of
species in stationary vs. mobile laboratories. Mobile
toxicity labs, on-site, have two advantages: (1) avoid-
ance of long-term sampling storage and (2) immediate
testing. All lab species identified previously are
suitable for mobile labs except: (1) some fish, de-
pending on age and species (and vulnerability during
transport) and (2) spawning stock.
In situ studies. Definition: In situ refers to an
organism immersed directly in the ambient water
enclosed in a screened or other type of chamber.
Problems with research-oriented species may be
magnified by in situ complications. Expected prob-
lems include general problems with egg and larval
stages (most cannot be used) and problems with
Xenopsis adult stage (air-breathing) animals.
Appropriate freshwater species include :
• Bluegill
• Striped bass
• Catfish
• Fathead minnows
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• ?White perch
Appropriate estuarine species include:
• Sheepshead
• Menldia
• Mysid shrimp
In observing adult stages in situ we won't necessarily
observe chronic toxicity; therefore we must be sure
that the effects observed are from contaminants and
not from the physical setting. Three factors can cause
problems: salinity, temperature, and food supply.
Suborganismal testing has potential for use in
ambient toxicity testing. Some tests may function as
early warnings, some as indicators of a specific
stressor, and some as indicators of classes of com-
pounds inducing reactions. However, not al] methods
are yet well-established. These methods must be
properly validated and documented before they are
applied to field conditions.
Types of Suborganismal tests:
• Immunological suppression
• Cytochrome P450 (a fairly good data base is
associated with this)
• Metallothioneins (these are good indicators of
metal regulation, especially Cu and Zn)
• RNA/DNA ratios in larval fish in feeding experi-
ments
Stress proteins (these offer a universal response to
stress, regardless of species; often single or a few
organisms are sufficient for testing).
Question: Can we use biomarkers in trend analysis?
Response: If we do so, we should use a suite of
responses rather than a single one.
Question: Is a reference toxicant test applicable to
biomarkers; are they used as such?
Response: We need to establish comparable databases
for all biomarkers. Future goals will include gene
studies on the source of protein synthesis and the
initiation of transcription.
Question: Should we use biomarker methods in the
pilot study when they are still in the research phase?
Response: We shouldn't discount them, because they
can indicate whether the system is getting better or
worse. They can act as a forensic approach to toxics
analysis. If we include them in the pilot study, then
both the specific and general information obtained
will be available for linkage to specific toxics.
Costs. We need to further address question #3
dealing with the costs. Maryland and Virginia will
provide us with the cost data associated with their
programs for regulatory methods (NPDES). No
definite estimates are possible with in situ testing;
they are very site-specific. If we are given the
parameters, then we can estimate.
Clarifications of category distinctions:
• Proven regulatory methods
• Chesapeake Bay-specific established methods
• Chesapeake Bay research methods
Water quality and contaminants data. We must
include the general water quality considerations, i.e.,
DO, salinity, temperature, etc. The in situ testing
allows for continuous measurements over long periods
of time. Testing considerations will be driven by/re-
stricted by the relative costs. Accurate testing will
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require background data review to narrow down the
number of toxicants to consider.
Comment: Toxitity should drive the types of analy-
ses done. We should be careful not to prejudge the
presence/absence of toxicants; we should allow the
testing to prove this.
TCP (inductively coupled plasma emission spectrosco-
py) analysis could be coupled with CBP (Chesapeake
Bay Program) water quality analysis approaches,
assuming that detection limits are appropriate for
ambient analysis.
We should emphasize the importance of the overall
coordination of investigators to avoid duplication of
testing efforts. We recommend panel review of
sample collection, perhaps through an RFP process:
• conducting sediment contaminant analyses with
water column
• contaminant analyses
• using composite sampling and grab sampling.
Comment on the recommendation in the Statement of
Purpose (p. 6), "Samples should be preserved and
analyzed only when biological test results are positive
for toxicity:" If levels of toxicants are found in the
system and organisms still do well, this is an impor-
tant aspect of the results and should be included.
The contaminants measured that show no toxicity are
important.
Question: Will a decision on the pilot study consider
the science equally with the socio-political consider-
ations?
Response: I don't think we're locked into the
proposed study areas. Steve Jordan mentioned
yesterday that the area suggested for the pilot study
was basically to promote discussion.
Water sampling
We need:
• Consistency in the in situ testing procedure to be
able to compare the responses of organisms.
• Lab analyses of grab samples are standard meth-
ods, but we will emphasize the use of composite
samplings.
Funding considerations
Point: We need to come up with realistic methodolo-
gies based on budget restrictions.
Response: Our primary concern is to determine the
best science to apply to the problem. Budgetary
considerations come after that. We should not focus
on the amount of money available but rather the
proper methodologies to be used.
Response: The number of sites has not been decided
and will depend on the funding available. Continual
efforts are being made to increase the monies avail-
able. The letter to EPA Administrator William Reilly
from 25 Bay-wide researchers made the point that the
presently authorized monies are not sufficient to
adequately answer the toxics questions at hand. We
may not know until September what money will be
available in October.
Suggestions:
Match the pilot study stations to already-estab-
lished Bay-wide monitoring program stations.
Exploit in situ techniques (for instance, simple
oyster "trees" some years ago yielded a lot of
information about vertical variations in both
concentrations and uptake of metals).
Incorporate histopathology, which can indicate
effects on health of Bay organisms. Sample
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indigenous fish and invertebrates within the
pollution gradient of the pilot study.
Presentation of risk assessment group's conclusions
was made by Ian Hartwell (see Risk Assessment
Summary).
Discussion
Comment: It seems that the community was empha-
sized in the risk assessment discussion rather than the
organism/species emphasis that was evident in the
water column discussion.
Response: Yes, because tests done on the organis-
mal/species level often are not done in a manner that
allows the data to be used for risk assessment. We
must be able to extrapolate this data to estimate the
integrity of the community.
Question: Did you address in your group the opera-
tional requirements of a pilot study, i.e., personnel,
expertise, and time?
Answer: We tended to focus more on a long-range
program. We decided it would take a number of
years and work-hours to establish program databases.
Further information to consider would include model-
ing links, which may require specialized people.
Question: Do you have an existing model available?
Answer: With the present pilot study restraints, there
are no good estimates of chemical exposure. If we
had exposure data or could generate data (which
would require a substantial amount of money), or if
we could collate for Chesapeake Bay data from other
data bases, then we could apply these to the estima-
tions of chemical exposures. We can't do risk
assessment without exposure data.
Question: What types of chemical exposure data do
you need?
Answer: We'd be looking for cause/effect-type
studies, including subcellular, suborganismal, and
whole organism information. It would be valuable to
be able to target areas of toxic concern (hot spots)
and be able to apply this data to a long-term ap-
proach.
Question: What are your abilities in predicting a risk
associated with a defined hot spot in the Bay system?
Answer: If we can measure the level of harm in a
species or group of species, then through cause and
effect modeling or extrapolation modeling we can
create a probability function that we can apply to the
community via extrapolation.
Comment: A number of existing programs already
have data from monitoring activities, and we should
draw from these for our considerations here.
Comment: EPA has asked CRC to collect all the
historical data available on Bay toxics, and the results
of this collection will be made available next year.
Notes on in situ selection
Livingston et al. (1986) used benthic "microcosms"
(colonization plates, sediment traps) to transfer
communities between clean and contaminated areas.
These experiments could be useful in in situ tests for
toxicity. Such experiments imply relatively long
exposures (probably 14-30 days).
Pratt et al. have transplanted artificial substrates in
streams, moving communities from reference sites to
"impacted" sites, and have measured loss of species,
decreased biomass, changes in nutrient content, and
nutrient transporting enzymes. This work is in review
(in part) with ASTM Aq. Tox. 13th Symposium.
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Questions Related to Sediment Toxicity Assays
Considering that the primary management need of the
Chesapeake Bay Program related to this topic area is
to identify sediments that are contaminated to the
point where living resources are impaired (either by
direct contact, ingestion, or fluxes of toxicants into
the water column), the following questions should be
addressed:
1. What routes of exposure should be assessed? (e.g.
ingestion by deposit-feeding fauna; direct contact
by epibenthic fauna; ingestion of suspended solids
by filter feeders; direct/indirect contact by fish;
bioaccumulation and food chain accumulation,
etc.)
2. What types of assays are currently available to
assess the effects of these routes of exposure?
3. What is the relative feasibility of these assays in
terms of costs and technical ease (i.e. time, train-
ing, and specialized facilities requirements) in a
large scale monitoring program?
4. What species should be selected for the assays?
(Consideration should be given to practicality,
cost, and sensitivity, as well as importance to and
representativeness of living resources in the Bay.)
5. What biological endpoints should be selected?
(They should be relatively rapid, sensitive, inter-
pretable in terms of risk to living resources, and,
hopefully, display continuity with "whole organ-
ism" and "suborganismal" assays being discussed
by other work groups for assessing ambient water
and effluents.)
6. How can sub-organismal measures be incorporated
into traditional acute or chronic toxicity tests?
How can results be interpreted in terms of impact
on living resources and their habitats?
7. What sort of experimental design(s) should be
adopted in a "full scale" assessment of the Bay?
(i.e. sites selected randomly, uniformly, or on a
stratified random basis; the number and definition
of "reference" or "control" sites; areas that should
be targeted for special effort; the role of a tiered
approach, etc.)
8. How can-the results of the assays be.integrated
with other monitoring data to "feed" into a Chesa-
peake Bay Risk Assessment Strategy?
a. What sort of chemical monitoring would be
needed to support this effort?
b. What sort of biological monitoring of in situ
living resources would be needed to support this
effort?
c. What other information would be required?
Workgroup Sessions on Sediment Toxicity
Assays
Convener: Ray Alden
Chair: Richard Peddicord
There was a general discussion about how to design
a strategy of sediment toxicity testing in the Potomac
Pilot study, the feasibility of such a study
given the budgetary constraints, and whether the
Potomac was a suitable place to develop testing
strategies applicable to the whole Chesapeake Bay.
Question: How do we use, set up, and apply these
tests to the Bay? (Question 7 was chosen as the
beginning point in discussion.)
The pilot program can be used to field-test both
standard and modified-standard techniques and to de-
velop a basis for recommending specific, more
chronic, estuarine-based techniques. Cost factors
must be assessed at the same time.
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Sediment quality criteria. It is agreed that sediment
quality criteria (SQC) are needed, but consensus is
lacking on bow they should be used and implemented.
Database development Development of SQC will
require a large database. A database should be
developed which will also support future application
of SQC and/or other sediment quality evaluation
techniques. Parameters for such a
database are presently being collected and observed,
but coordination is needed in the gathering of this in-
formation. A database can be developed that will aid
in developing SQC. Some work in the Elizabeth
River is being done to relate toxicity tests to SQC.
Consensus: We need to collect supporting data for
SQC when doing toxicity tests and chemical analysis,
to the extent that we know what these data are. W
e need to compile not only the relevant sediment
chemistry, benthic data, and toxicity data, but also
data on grain size, outfall locations, sources of
contamination, etc., and use this information to guide
the Program.
Pilot study siting
Question: How did the Potomac River get chosen
for the pilot study?
Response: The research was already done on it, it
spanned political jurisdictions, and it was an integrat-
ed estuarine system (encompassing tidal freshwater
through mesohaline water).
The question arises whether sediment toxicity is
causing problems for living resources. If not, then
what is the use of studying sediment toxicity? Living
resources are the focus of public attention. For
instance, the striped bass is still in trouble in the
Potomac, but there is public pressure to open
the fishing rights again, because the fish larval index
has improved slightly.
Question: If the Potomac is relatively clean then why
look for sediment toxicity there? Not much sediment
data exists. It would make sense, if the Potomac is
studied, to include a comparison of the Elizabeth
River to the Potomac: worst case compared to rela-
tively clean. This would also provide verification that
the methods used in the Potomac do work in a site
where we already know there is toxicity.
Consensus: A positive control should be established,
perhaps outside the Potomac system, to demonstrate
that the tests being run do work.
Comment: The budget for the entire pilot program is
$250,000, for all aspects of the study.
Question: What is the relation of sediment toxicity to
the striped bass fishery?
Response: Answering this question probably involves
looking at other resources, i.e., benthic species.
Question: Do we know what tests and present body
of knowledge exist for the quality of Potomac sedi-
ments?
Response: The D.C. Department of Environmental
Control has recently conducted benthic community
structure analyses for several sites in the upper tidal
Potomac and Anacostia Rivers. These studies showed
most of the Potomac sites to have fair health indices,
with some in the moderately good range. All sites in
the Anacostia reflected poor indices. A continuing
study of Anacostia sediment toxicity has found
considerable sediment toxicity, particularly at the
mouth of the Anacostia. The recently improved
health of the upper Potomac has been partly attributed
to the water filtering capacity of the clam, Corbicula,
and the return of SAV, which has encouraged the re-
turn of several fish species.
The Great Lakes Program used a suite of tests to
determine areas of concern as part of the ARCS
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program, and then employed bioassays to find sedi-
ments which were better or worse in quality. These
tests included:
• sediment grain size fractions
* organic carbon
• solvent extractables
• organically-bound chlorine, bromine, and iodine
• inductively coupled plasma (ICP) analysis of
selected metals
• Microtox bacterial luminescence assays (perhaps
not appropriate to the Bay given the narrow
salinity range restrictions).
Perhaps we should be looking for new and innovative
measures that will give us more complete information.
Many of the standard tests applied to sediments have
historically been adapted from old water quality tests.
There is a real need to develop new tests aimed
specifically at sediment quality testing needs.
Identification of sites in the Potomac. We need to
use existing information to identify areas where
possible sediment toxic effects might be. We need
to do toxicity testing on both the very dirty and the
clean sites, as well as the gray areas in between. We
need both positive and negative reference sites so that
we can establish the outermost ranges of both natural
survival and the anticipated biological endpoints.
Establishing such a gradient, encompassing the range
of toxicity, would also allow us to establish or mea-
sure the relative sensitivity of the toxicity tests.
Comment: The charge is to determine the effects of
toxics on living resources of the Bay.
Comment: We should start with benthic community
structures to identify what areas are of concern.
Since this is a demonstration project, we will have to
show responses across a contaminant gradient.
Question: What are we going to set as a biological
impact level? What level of reduction in survival is
statistically significant?
If the sediment bioassays (acute tests) tell us that the
benthic area is disturbed, we still can't make large
leaps from sediment bioassays to living resource risk
assessment. But we can make the connection from
sediment tests to benthic community structure effects.
Variability. Geologists admit that there is large
variability in benthic community data. Many factors
can cause a disturbance; it is hard to attribute
a change to one or two factors. The best approach is
to look at all three components, i.e. sediment chemis-
try, toxicity, and benthic community, to get a
handle on sediment quality.
It is important to identify what we will consider as a
statistically significant impact. The protocols should
specify that design and sampling schemes be set to
get results that show xx% response that signifies a
significant difference.
There are two types of variability: (1) natural variabil-
ity in sediment quality that can be partially overcome
by compositing sediment samples from a site, and (2)
variability due to the precision level of the test
method and variability of species. Amphipod tests
show that differences of 15% are usually significant
We need a more qualitative examination of benthic
community structure before we can assess the impor-
tance of the variability that is observed.
Although funding constraints may limit how much
sampling can be done, the problem with using one
sample per site is that you don't learn anything about
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within-site variability. We need to determine the
variability in endpoints and ecological conditions; this
would require a suite of tests in one area.
We should use a fairly quick, cheap screening tool,
then use the toxicity tests. The reproducibility and
power of the test should be kept in mind when
interpreting results. Data bases are available for
making estimates on the level of replication that will
give differences.
Question: Should within-site variability be our focus?
The pilot parameters will determine the site, species,
and variability. The power of the test must be re-
corded and verified during study.
Once we have done sampling then we can get a
handle on the variability. We should use species that
have demonstrated sensitivity; we should test whole
sediments and then determine which tests can be
applied to the Bay.
Question 2: "What types of assays are currently
available to assess the effects of these routes of
' xposure?" Basic recommendations are to use whole
sediment methods and to test species that have some
existing database.
Possible tests.
Standard acute
10-day amphipod test for screening (there are
amphipods, Lepidactylus and Leptocheirus, for
example, that should be used and developed).
The basic screening tests that have been fairly cost
effective for the Great Lakes.
Suggestions:
• Use biological tests (benthic community structure)
first to screen, then chemistry testing and sedi-
ment bioassays.
• Use a suite of test species and methods.
• Use infaunal species rather than epibenthic ones.
• Demonstrate the utility of an amphipod test for the
Bay using existing standard methods and species
indigenous to the Chesapeake Bay.
• Develop and field-validate tests for chronic and
population effects of toxic sediments using Chesa-
peake Bay indigenous species.
The primary goal of the Chesapeake Bay Agreement
is to protect living resources. However, our knowl-
edge of the impacts of toxics in the Bay is weak.
This workshop should provide the guidelines for
toxics assessment. The drafters of the Agreement did
not know a great deal about sediment toxicity.
A question we haven't addressed is freshwater toxici-
ty testing. The relative sensitivity of species across
salinity gradients is important. We should include
this in our recommendations.
Selection of species. Discussion on standard tests and
tests in development that are appropriate for the
various salinity levels of the Chesapeake Bay is
summarized in a table (see Final Plenary).
A test showing promise, but still in development, is
one using Sago pondweed (Potomageton pectinatus).
It was at one time the primary food for many migra-
tory birds, but after being historically present it has
disappeared. Using tissue culture techniques, it can
be produced year-round. The test takes about 4-6
weeks, as it measures mortality over time; several
endpoints can be used. It is of interest because it
represents a problem group (SAV), and because it
represents the primary producer level.
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Standard protocols must be set forth for sediment
toxicity testing for the Chesapeake Bay and should be
required for use by all pilot programs and final
program investigators.
Recommendations. Before testing begins we must
have a quality assurance/quality control plan and
develop standard operating procedures for the Chesa-
peake Bay. These should address the following:
• sediment collection and storage
• extraction procedures
• test procedures.
Bioaccumulation
Question: How should bioaccumulation be considered
and evaluated?
If we use fish monitoring, we should try to select a
species that has been shown to accumulate through a
sediment source. However, routine monitoring of
bioaccumulation may not be pertinent to the charge of
the program as a threat to living resources. There
should be a demonstrated problem due to bioaccumul-
ation in an area before we say it's necessary to study
it. Widespread studies would not be cost-effective.
Both Maryland and Virginia will continue to do
bioaccumulation/tissue residue monitoring focused on
edible fish and shellfish since it's such a huge eco-
nomic issue. If these programs find a problem, then
we will have a justification for looking further into it.
We can examine the living resources and try to
identify whether the toxics are coming from the
sediments or the water.
We should define an area of sediment sampling and
limit it to the 10-year flood plain. Some tidal plains
and marshes have some of the worst problems with
toxic sediment.
Sub-aquatic, water-saturated sediment must be consid-
ered separately from soils.
Protocols for sediment depth sample. Based on the
specific hypotheses/questions being stated/asked, the
depth of sediment collected for toxicity testing must
be clearly defined before implementation of the pilot
program. Factors to be considered include focus on
recently deposited sediments, average depth for
benthic infauna (10-25 cm), depth of dredging (depen-
dent on site), and a historical review of sediment
toxicity (core sectioned by depth according to local
depositional patterns).
The population risk assessment group has tried to
come up with a ranking of sites, with sediment
toxicity as one factor.
Question 6 — How can suborganismal measures be
incorporated into sediment toxicity work?. The
workshop participants working in this field will
incorporate some material from the SETAC meeting
into this report.
We could use pore water tests in conjunction with
cross-media testing. We should give priority to cross-
media methods. Sub-organismal methods are best
tied in with chronic tests. Metabolic enzymes will
help in the "fingerprinting" of sources of toxicants.
Some genetic work has been done in Michigan with
mollusks, and although problems remain in extrapolat-
ing suborganismal effects all the way to population
risk assessment, the work has potential. It could be
useful to poll the medical community for ideas on
extrapolation from sub-organismal to environmental
risk assessment.
Sublethal effects may be good tools to assess caus-
ative factors of toxicants, although there are prob-
lems of covariance. Sediment mixtures of contami-
nants that are separated do not always behave the
same as the original mixtures, due to additive effects.
The attribution of toxicity to a specific toxicant by
means of these tests may prove easier in the water
column than in the sediments.
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Sublethal effects can also be used to track subtle
changes in organisms. A sublethal effect is an early
warning signal that can be connected to growth and
reproduction effects.
There are two classes of responses: (1) a stress-
protein response to general stress and (2) other
responses that can be used for specifically identifying
a responsible contaminant. These two classes of
response may be used in a tiered system. An assay at
tier 1, e.g., a stress-protein assay, will determine an
effect, and an assay at tier 2, e.g., metallothionein or
cytochrome P450, will identify what is causing it.
We are reaching the point where the answers to
specific toxicity can be teased out, but it is somewhat
tricky.
The economics of this kind of tiered approach are
good, as these assays are much cheaper than routine
sediment chemical analysis. Some labs can run these
tests on a routine basis.
There is need for an entire suite of tests, not just the
standard amphipod assays. However, polychaetes
seem to be less sensitive, and whole-sediment expo-
sure tests are better developed for amphipods. We
are hampered by the fact that appropriate tests for
sediment in other phyla haven't been thoroughly
developed and tested, and we are working on a short
time budget.
For risk assessment, endpoints of mortality, fecundity,
and growth and bioenergetic measures are most help-
ful. The relative sensitivities of species and endpoints
are important for statistically generated models. From
a managerial standpoint, looking at species representa-
tive of the Bay is important. A manager would want
to relate tests to Callinectes, Crassostrea, and impor-
tant fish species.
The exposure of animals is difficult to assess in
species you aren't familiar with. Also, the animals
chosen have to be maintained in labs. This necessity
will restrict the species selected to those that are
robust enough for this handling, yet also meet all the
other criteria of sensitivities needed for sediment
testing. Since the relative sensitivities of benthic
invertebrates aren't known, then toxicity tests should
be coupled with benthic community analysis.
There is need for a gradient of contamination in the
pilot testing, and many have doubts whether the Poto-
mac offers this gradient. The problem of finding a
true uncontaminated or no-effects reference site(s)
will be difficult but also critical.
A standard reference sediment may be needed so that
all toxicity tests are run with the same reference. The
selection of reference sites will depend on the objec-
tives of the Program. If the objectives are to assess
relative contamination within the Bay, then this
information will evolve from data collected at all sites
tested. Strict hypothesis testing would require more
stringent reference definition.
Recommendations
Suborganismal endpoints should be developed.
However, they are not ready for immediate use in
this Program; first they must be directly related to
whole-organism effects.
The Program needs specific, focused objectives
and testable hypotheses before the question of
reference sediment and sites can be adequately
addressed.
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Appendix B
Workshop Participants
Methodologies for Whole Organism Toxic-
ity Testing
Plenary Speakers:
Mr. Steve Schimmel
Environmental Research Laboratory
U. S. Environmental Protection Agency
Dr. Jeffrey Black
Graduate Center for Toxicology
University of Kentucky
Workgroup Chair:
Mr. Steve Schimmel
Environmental Research Laboratory
U. S. Environmental Protection Agency
Convener:
Mr. Lenwood Hall
Applied Physics Laboratory
The Johns Hopkins University
Workgroup Participants:
Dr. Brian Bradley
University of Maryland
Dept. of Biological Science
Dr. Arthur Butt
Virginia Water Control Board
Dr. John Cooney
Battelle Memorial Institute
Dr. David W. Engel
NOAA/NMFS
SE Fisheries Center
Beaufort Lab
Dr. Jay Gooch
Chesapeake Biological Laboratory
University of Maryland
Dr. David Gruber
Biological Monitoring Inc.
Mr. George Kennedy
Hampton Roads Sanitation District
Dr. Arthur Ott
Susquehanna River Basin Commission
Mr. William Pfeifle
Virginia Water Control Board
Dr. John Pritchard
Laboratory of Pharmacology-NIEHS
Dr. G. Roesijadi
University of Maryland
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Chesapeake Biological Laboratory
Mr. Keith Sappington
Maryland Dept. of the Environment
Mr. John Veil
Maryland Dept. of the Environment
Dr. Dave Wright
Chesapeake Biological Laboratory
University of Maryland
Recorder:
National Oceanic and Atmospheric Administration
Washington, DC
Ms. Sherri Clark
Virginia Water Control Board
Dr. Jim Hemming
US Fish & Wildlife Service
North Carolina State University
Dr. Michael I. Mac
Great Lakes Fishery Lab
U. S. Fish & Wildlife Service
Ms. Robin Laird
Chesapeake Bay Liaison Office
Sediment Toxicity Assays
Plenary Speakers:
Dr. K. John Scott
Environmental Research Laboratory
U. S. Environmental Protection Agency
Dr. Chris Zarba
U.S. Environmental Protection Agency
Workgroup Chair:
Dr. Richard Peddicord
E A Engineering Science and Technology
Convener:
Dr. Ray Alden
Appb'ed Marine Research Laboratory
Old Dominion University
Workgroup Participants:
Dr. H. Suzanne Bolton
Mr. Eh' Reinharz
Maryland Department of the Environment
Ms. Debra Trent
Virginia Water Control Board
Recorder:
Ms. Kim Warner
Chesapeake Biological Laboratory
University of Maryland
Population Risk Assessments Based on Toxicity
Testing
Plenary Speakers:
Mr. Donald Rodier
U. S. EPA, Office of Toxic Substances
Dr. Glenn Suter II
Oak Ridge National Laboratory
Workgroup Chair:
and Convener:
Dr. Ian Hartwell
Horn Point Environmental Laboratories
University of Maryland
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Workgroup Participants:
Mr. Charley Banks
"Virginia Water Control Board
Dr. Katherine Farrell
Environmental Science and Health
Maryland Department of the Environment
Dr. Yacov Haimes
Department of System Engineering
University of Virginia
Dr. Kenneth Jenkins
CBR, Inc.
Ms. Gail MacKiernan
Sea Grant Program
University of Maryland
Dr. Robert Otto
Otto & Associates
Dr. Kenneth Perez
EPA, ERL, Narragansett
Mr. Mark Richards
Virginia Water Control Board
Dr. Robert Ulanowicz
Chesapeake Biological Laboratory
University of Maryland
Recorder:
Ms. Jackie Savitz
Chesapeake Biological Laboratory
University of Maryland
Methodologies for Sub-organismal Toxici-
ty Testing
Plenary Speakers:
Dr. G. Roesijadi
Chesapeake Biological Laboratory
University of Maryland
Dr. Jay Gooch
Chesapeake Biological Laboratory
University of Maryland
Conveners:
Dr. Jay Gooch
Chesapeake Biological Laboratory
University of Maryland
Dr. Peter Van Veld
Virginia Institute of Marine Science
Additional Workgroup Participants
(Day 1 Plenary and Workgroup "floaters")
Mr. Dan Audet
Chesapeake Bay Estuary Program
U. S. F. W. S.
Mr. Richard Batiuk
U. S. Environmental Protection Agency
Chesapeake Bay Liaison Office
Mr. Mark Bundy
Maryland Dept. of Natural Resources
Dr. Dennis Burton
The Johns Hopkins University
Dr. William Busey
Experimental Pathology Laboratories
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Ms. Liz Conner
University of Maryland
Dr. Eugene Cronin
Private Consultant
Dr. Rex D'Agostino
University Micro Reference Lab
Dr. Tom Dillon
Waterways Experiment Station
U. S. Army Corps of Engineers
Ms. Phyllis Frere
PEPCO-Hallowing Point Lab
Ms. Elaine Friebele
Interstate Comm. on the Potomac River Basin
Ms. Mary Jo Garreis
Maryland Dept. of the Environment
Ms. Bess Gillelan
Estuarine Programs Office
Ms. Eileen Hamilton
Chesapeake Biological Laboratory
University of Maryland
Mr. George Harmon
Maryland Dept. of the Environment
Mr. Carlton Haywood
Interstate Comm. on the Potomac River Basin
Ms. Paula F. P. Henry
USFWS-Patuxent Wildlife Research Center
Mr. Lance Himmelberger
Penna. Dept. of Environmental Resources
Mr. Jerrald HolloweU
Susquehanna River Basin Commission
Mr. M. Paul Jackson
Martel Laboratory Services
Dr. Margaret O. James
Department of Medicinal Chemistry
University of Florida
Ms. Betsy Johnson
Water Hygiene Branch
District of Columbia Government
Mr. David Jordahl
Maryland Dept. of Natural Resources
Dr. Stephen Jordan
Maryland Dept. of Natural Resources
Dr. Andy Kane
School of Medicine
University of Maryland
Mr. Stuart Lehman
Chesapeake Bay Foundation
Ms. Laura Lower
Virginia Council on the Environment
Ms. Patmarie Maher
Estuarine Studies Office
Dr. Joseph Mihursky
Chesapeake Research Consortium
University of Maryland
Dr. Jack Plimmer
USDA, Agricultural Research Service
Mr. Alan E. Pollock
Virginia Water Control Board
Dr. Richard Pratt
School of Forest Resources
Pennsylvania State University
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Mr. David Pyoas
Maryland Dept. of Natural Resources
Dr. Andrew Robertson, Chief
Ocean Assessments Division
NOAA/NOS, N/OMA3
Mr. John Slowikowski
Industrial Discharge System
Maryland Dept. of the Environment
Dr. Roland C. Steiner
Interstate Comm. on the Potomac River Basin
Ms. Cynthia Stenger
Maryland Dept. of Natural Resources
Mr. James T. Ulanoski
Penna. Dept. of Environmental Resources
Dr. Wolfgang K. Vogelbein
Virginia Institute of Marine Science
Dr. Clarence Wade
University of the District of Columbia
Dr. Lloyd Wolfinbarger, Jr.
Center for Biotechnology
Old Dominion University
Dr. Chris Zarba
U. S. Environmental Protection Agency
Workshop Staff
(All staff were employed by the Chesapeake Research
Consortium.)
Ms. Karen McDonald
CRC Project Manager
Ms. Dana Flanders
Ms. Pam Owens
Ms. Cindy Corlett
Ms. Elizabeth Krome
Ms. Deb Young
Ms. Elizabeth Egeli
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Appendix C
Bioassay Capabilities Survey Results
The information for this appendix did not come directly from the workshop, but was collected
by workshop staff and reflects the recommendations made by the workshop steering committee.
An introduction follows which explains the purpose of this addition and the process by which
it was developed. A separate page of acknowledgements is included as well.
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ACKNOWLEDGEMENTS
Thanks are expressed to Cindy Corlett, Pam Owens, and Jackie Savitz for their coordinated
efforts to develop the questionnaire, contact labs, enter response data, develop a compilation
format, and edit the final product. Gratitude is also extended to the representatives from the labs
listed below who gave their time to complete the questionnaire.
Participating Laboratories
Academy of Natural Sciences
Applied Marine Research Laboratory
Aqua Survey, Inc.
Biological Monitoring, Inc.
Bionetics Corporation, Analytical Laboratories
Division
Center for Environmental Studies
Chesapeake Biological Laboratory
Coastal Bioanalysts, Inc.
Commonwealth Laboratory, Inc.
EA Engineering, Science, and Technology, Inc.
Environmental Laboratories, Inc.
Environmental Resources Management, Inc.
Environmental Systems Service, Ltd.
Experimental Pathology Laboratories, Inc.
Free-Col Laboratories, Inc.
Hampton Roads Sanitation District
Horn Point Environmental Laboratory
James R. Reed and Associates
Johns Hopkins University, Applied Physics Lab
Malcolm Pimie, Inc.
Olver Incorporated
Patuxent Wildlife Research Center
Riverside Laboratories
Technical Testing Laboratories
University of Maryland, Agricultural Experiment
Station
University of Maryland at Baltimore, Aquatic Toxi-
cology Facility
University of Maryland, Baltimore County
Versar, Inc. ESM Operations
Virginia Institute of Marine Science
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INTRODUCTION
About the survey
This appendix is the product of a survey conducted by
the Chesapeake Research Consortium, Inc. (CRC)
September through December of 1989. This survey
was conducted to determine the capabilities available
for conducting bioassays within the Chesapeake Bay
watershed. A questionnaire was developed and sent
to all laboratories in the region who were identified as
possible candidates. Fifty-one laboratories in Virgin-
ia, Maryland and Pennsylvania were sent question-
naires and explanatory cover letters. Of these, 29
laboratories participated in the survey: the responses
from their completed survey forms comprise this
appendix.
The remaining laboratories did not respond for a
variety of reasons. Some questionnaires were re-
turned to CRC by the post office. Some lab manag-
ers reported that they did not conduct applicable tests.
Others simply did not respond. A listing of these
laboratories and their response status is available from
the Chesapeake Research Consortium upon request.
About this compilation
This booklet is divided into two sections. The first is
a series of one- to two- page entries which are ar-
ranged alphabetically by lab and begin on page 47.
These entries contain general information about the
laboratories such as lab address, a contact person,
types of bioassays conducted, field sampling capabili-
ties, accessible analytical chemistry equipment,
computers and software commonly used,
staff available, and data on respective quality assur-
ance/quality control programs. The format of these
pages follows that of the original questionnaire
closely. Likewise, care has been taken to enter the
labs' responses as closely as possible, notwithstanding
necessary editorial changes.
The second section is arranged by type of bioassay,
giving specific information about the bioassays con-
ducted at each lab; it begins on page 205. There is a
table for each of the following types of bioassays:
acute fish, acute invertebrate, bacterial, biochemical
("biomarkers"), chronic fish, chronic invertebrate,
plant (algal), and sediment. Entries within each table
are listed alphabetically by lab and include informa-
tion on species, test conditions, organism's life stage,
test length, salinity, lab's holding capacity, test turn-
around time, and the number of bioassays each lab
can run simultaneously.
Future assessment
It is our hope that these results will be utilized both
as a catalog of the services available for contracting
and as a data compilation from which planners can
determine areas to target for future development.
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Academy of Natural Sciences
James Sanders Testing site: Benedict Estuarine Research Lab.
Benedict Estuarine Research Laboratory Benedict, Maryland 20612
Benedict, Maryland 20612 Hours: 8:00 AM - 4:00 PM
301-274-3134
FAX 215-299-1199
Bioassays Conducted
Types of bioassays: The Academy of Natural Sciences does not do conventional bioassays. However, much of their
research is devoted to assessing the impact of low levels of toxics on natural estuarine communities. Many of these
studies result in techniques useable for toxicity assessment. For example, they maintain a microcosm facility that
enables them to investigate toxic impact to phytoplankton communities. They also routinely maintain 30 or so algal
species in culture, which could be utilized for assays. (See tables for description of exact test capabilities.)
Future plans: The Academy will continue with the same areas of research.
Field sampling capabilities: sampling gear for water column, organisms and sediments; variety of vessels, 20-45
feet
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty:
- trace elements; particularly organ metalloids
- organics; analyzed by sister lab in Philadelphia
Major equipment available:
electron microscopy: none reported
spectrophotometry: 2 AA's for elements in Benedict
MS
GC/LC MS in Philadelphia
chromatography: GC in Benedict
GC, LC's in Philadelphia
Additional Resources
Computerized information retrieval services available: DIALOG
Computers and software available:
- usual mix of personal computers and software
- VAX 11/730 running VMS and SAS
Staff available: 6 Ph.D.- level senior scientists
20 B.S./M.S. support staff
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Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? Yes
(Only in compliance at Philadelphia Lab.)
Follow study protocols? N/A Have a complete set of Standard Operating Procedures? N/A
Archive facility for the data generated? N/A
Computer-generated data? N/A Are these computer systems validated? N/A
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Applied Marine Research Laboratory
Old Dominion University Testing site: AMRL
Dr. Raymond W. Alden, III Research East Building
4401 Powhatan Avenue Center for Biotechnology
Norfolk, Virginia 23529-0456 Dept. of Biological Sciences
804-683-4195 Mills Godwin Bldg.
FAX 804-683-5293 Old Dominion University
Norfolk, Virginia 23529
Hours: 24 hrs./day
Bioassays Conducted
Types of bioassays: The Applied Marine Research Lab (AMRL) conducts acute and chronic bioassays on fish and
invertebrates as well as a suite of sediment-organism bioassays. (See tables for descriptions of exact test
capabilities.)
Other bioassay work: Toxicological and biotechnological assays of various sorts are now being developed at ODU.
Cell culture and microbial cultures are being developed for extensive screening activities; histopathological,
behavioral, genetic and teratogenic assessment are being developed for testing subtle chronic effects of toxicants on
aquatic organisms.
Future plans: AMRL and the Center for Biotechnology plan to continue development of a toxicology emphasis,
developing sensitive, cost-effective toxicity tests capable of high capacity sample through-put. Flow-through and
mobile/on-site capabilities are also being developed.
Field sampling capabilities:
- full range of collection protocols and gear for obtaining organisms, water, sediment, and wastewater samples
- fleet of research vessels from small (10-20 ft.) and mid-size (20-40 ft.) boats to a 65 ft. vessel, the R/V Holton
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: Laboratory sections that specialize in aquatic toxicity; organic contaminant
analysis; metal analysis; water quality analysis; field environmental assessment; biological monitoring (fish, benthos,
phytoplankton, zooplankton) and research; biotechnology applied to environmental toxicity; and fisheries modeling
emphasizing survival and growth studies of juvenile fishes.
Major equipment available:
electron microscopy: JE01, 100CXII SEM/TEM with Kevex Delta-1 x-ray
microanalysis system
Cambridge S100 SEM
spectrophotometry: 2 Finnegan quadrapole GC/MS
2 Perkin-Elmer 5100 AA spec.'s with graphite furnaces
1 Thermo-Jarrell Ash video 12 AA spec's with graphite furnace
1 Appb'ed Research Laboratories 3410 ICAP
1 Perkin-Elmer Model 552 dual beam UV/VIS spectrophotometer
1 Perkin-Elmer Lambda I UV/VIS single beam spectrophotometer
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2-channel SCI Autoanalyzer system
chromatography: 4 automated GC systems with detectors (3 FID, 3 RCD, 1 FPP, 1 PID)
1 Waters automated HPLC system (UV and fluorescence)
1 GPC system
1 Carlo-Erba NA 1500 carbon/nitrogen analyzer
1 OI carbon analyzer
Additional Resources
Computerized information retrieval services available:
- Aquatic Biology and Fisheries Abstracts on CD Applied
- Agricultural & Biological Abstracts on CD Bitnet TOXNET
Computers and software available:
- IBM 3090: SAS on CMS, Pascal, FORTRAN (w/IMSL)
- IBM/PC's and PC clones: PCSAS, Turbo Pascal, Harvard Graphics, Statgraphics
Staff available:
- approximately 50 faculty members (Ph.D. level) working in various areas of marine research at ODU
- 8 senior AMRL staff (M.S.-level supervisor/managerial)
- approximately 30 full-time technicians with degress in oceanography, biology, chemistry, geology and
environmental health
- numerous graduate research assistants and undergraduate research assistants (20-30)
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989?
GLP requirements are being reviewed; full compliance is expected within 6-12 months; QA/QC protocols are ap-
proved by USEPA Bay Program VWCB Safe Drinking Water Program; and independent review (EA) is conducted.
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems validated? Yes
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Aqua Survey, Inc.
Ken Hayes Testing site: Aqua Survey, Inc.
499 Point Breeze Road 499 Point Breeze Road
Flemington, New Jersey 08822 Fletnington, New Jersey 08822
201-788-8700 Hours: 8:00 AM - 5:00 PM
FAX 201-788-9165
Bioassays Conducted
Types of bioassays: Aqua Survey, Inc. (ASI) conducts acute and chronic bioassays using freshwater, estuarine, and
marine species of fish, invertebrates, and algae. They also conduct toxicity reduction bioassays and ocean disposal,
oil dispersant, and Microtox tests. Some tests may be run in the mobile lab or in situ.
Other bioassay work: ASI is equipped with three mobile labs, 14 diluter systems, and has extensive acute, chronic,
bioaccumulation (sediments) and TRE
experience.
Future plans: ASI is in the process of patenting a procedure that can be utilized to determine rate and level of
toxicity to mixed microorganism populations.
Field sampling capabilities: grab samplers, composite samplers, piston corer, Ekman dredge, Alpha bottles,
Kemmerer sampler, zooplankton nets, epibenthic sled, seines
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: wet chemistry
Major equipment available:
electron microscopy: none reported
spectrophotometry: none reported
chromatography: none reported
Additional Resources
Computerized information retrieval services available: none reported
Computers and software available: several PC's
Staff available: 1 Ph.D.; 1 M.S.; 7 B.S.'s; 1 B.A.; 1 A.S.; 2 with degrees in progress
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? Yes
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Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems validated? Yes
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Biological Monitoring, Inc.
Mark Collins, Laboratory Manager Testing site: Biological Monitoring, Inc.
P.O. Box 184 Route 460 South
Blacksburg, Virginia 24063 Blacksburg, Virginia 24060
703-953-2821 Hours: 8:30 AM - 5:30 PM
FAX 703-382-6090 7 days/week
Bioassays Conducted
Types of bioassays: Biological Monitoring, Inc. (BMI) conducts acute and chronic bioassays on fish, invertebrates,
and some algae. (See tables for description of exact test capabilities.)
Other bioassay work: BMI also works with the following species under most test and flow conditions: small mouth
bass, striped bass, channel catfish, mummichog, mayflies, stoneflies, caddisflies, amphipods, water penny, blue
mussell. Most of BMTs experience with sediments has been testing of elutriates (freshwater and marine). Solid
phase studies have been performed in-house but not for contract.
Future plans: BMI plans to expand marine and estuarine culturing and testing capabilities and to increase solid phase
sediment testing capabilities.
Field sampling capabilities: 16" outboard motorboat, Kemmerer bottle, Ponar dredge, Surber sampler, kick-nets,
plankton nets, field water quality meters, phytometers; experience with freshwater and estuarine biological and
chemical surveys
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: standard physiochemical parameters including DO, pH, conductivity, salinity,
temperature, alkalinity, hardness, chlorine
Major equipment available:
electron microscopy: none in house; handled through sub-contract
spectrophotometry: none in house; handled through sub-contract
chromatography: none reported
Additional Resources
Computerized information retrieval services available: Chemical Abstracts, Biological Abstracts, Toxline, Toxnet,
AQUIRE
Computers and software available: IBM PC/AT's; IBM PS/2 Model 30/286; Toxstat and data system; EPA toxicity
test data analysis programs; WordPerfect; Harvard Graphics; Lotus 1-2-3; dBase III; IBM mainframe at Virginia
Tech;SAS
Staff available: full-time: 3 Ph.D.'s; 7 B.S.'s; 2 A.S.'s
part-time: B.S.; 5 students; 1 electrical engineer
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Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and F1FRA of August 17, 1989? Yes
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems validated? Yes
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Bionetics Corporation, Analytical Laboratories Division
Peter T. Pohorence Testing site: Bionetics Analytical Laboratories
20-A Research Drive 20-A Research Drive
Hampton, Virginia 23666 Hampton, Virginia 23666
1-800-476-5548 Hours: 8:00 AM - 5:00 PM
804-865-0880 FAX 804-865-7597 Monday - Friday
Bioassays Conducted
Types of bioassays: Through a joint association with Coastal Bioanalysts of Gloucester Point, VA, Bionetics is able
to conduct acute and chronic bioassays of both fish and invertebrates. They also conduct algal growth tests, Microtox
tests, Ames mutagenicity tests and others. Note that bioassay testing is not done by Bionetics staff; they only
conduct chemical testing. (See tables for complete descriptions of testing capabilities.)
Other bioassay work: Through a joint association with Coastal Bioanalysts, Bionetics offers in-stream impact studies:
freshwater, marine, benthic, nekton, phytoplankton, zooplankton, ambient toxicity. They also conduct municipal and
industrial Toxicity Reduction Evaluations and develop site-specific water quality criteria/standards.
Future plans: Through a joint association with Coastal Bioanalysts, Bionetics expects to increase the size of its
physical plant within the next two years - modifying to permit freshwater and marine flow-through/bioconcentration
studies. Increased capacity for static/static renewal testing is also expected within a year. Another possibility is
expansion to mobilize lab capabilities.
Field sampling capabilities: 4 24-hour discrete/composite 1500 field samplers; assorted grab sample apperatus; seine
nets. Core samplers, Ponar, Smith/Mac, Peterson, Ekman, Surber Samplers and plankton nets are available by lease.
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: inorganic, metals, and organic <
organic chemistry
Major equipment available:
electron microscopy: none
spectrophotometry: 1 VG Trio GC/MS
1 Perkin-Elmer 3939 with graphite and auto sampler
1 IL S-12 with graphite
1 J-Y 24 Sequential ICP with auto sampler
chromatography: 3 Varian GC's: (2) Model 3300, (1) Model 3400 with auto sampler
1 Waters GC with auto sampler
3 Tekmar Purge and Traps
Detectors available: 2 FID, 4 BCD, 2 PIO, 1 HALL, 1 FPD
Additional Resources
Computerized information retrieval services available: GC/MS database and library search for 48,000 compounds;
GC data retrieval system; ICP data retrieval system; DIALOG availability possible
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Computers and software available: CLP program data packages for GC/MS, GC, ICP. Novell Data System with
12 workstations. Programs: dBase, Lotus, WordPerfect. (See also Coastal Bioanalysts.)
Staff available: full-time: 1 Ph.D.; 2 M.B.A.'s; 8 B.S.'s; 1 A.A.; 10 non-degree
part-time: 11 non-degree
(See also Coastal Bioanalysts.)
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? Yes
Follow study protocols? Yes* Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems validated? Yes
* Coastal Bioanalysts — Protocol for effluent biomonitoring has been submitted and approved by Virginia Water
Control Board. SOP'S submitted to Maryland to date have been approved.
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Center for Environmental Studies
Virginia Polytechnic and State University
David R. Orvos Testing site: VPI&SU
Derring 1020 Derring labs 1027,1027A,1014A,
(VPI&SU) 1020A, 2006
Blacksburg, Virginia 24061 Blacksburg, Virginia 24061
703-231-5538 Hours: 8:00 AM - 5:00 PM
FAX 703-231-9307 (Ecosystem Simulation Labs have
no regular hours.)
Bioassays Conducted
Types of bioassays: The Center for Environmental Studies (CES) is capable of almost any bioassay as prescribed
by the contractor.
Future plans: They plan to continue as at present.
Field sampling capabilities: They routinely collect from lakes, streams, wetlands, and rivers.
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: routine water chemistry; soil and wetlands analyses on site
Major equipment available:
electron microscopy: Both TEM and SEM at 2 sites on campus; includes EDAX.
spectrophotometry: AA, UV/VIS on site (CES labs)
MS/GC, ICP on campus
chromatography: LC in CES lab
GC, MS/GC on campus
Additional Resources
Computerized information retrieval services available: All commonly used ones.
Computers and software available: IBM and compatibles; 80288-based; Lotus, Word, etc.;
IBM 3090 mainframe with SAS, Script, etc.
Staff available: 5 full-time Ph.D.'s; 8 graduate students; 1 research specialist
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? not reported
Follow study protocols? Yes
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Have a complete set of Standard Operating Procedures? Presently completing
Archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems validated? Mainframe, yes
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Chesapeake Biological Laboratory
University of Maryland, Center for
Environmental and Estuarine Studies Testing site: Chesapeake Biological Lab
Jay W. Gooch UMCEES
P.O. Box 38 Solomons, Maryland 20688
Solomons, Maryland 20688 Hours: all
301-326^281
FAX 301-326-6342
Bioassays Conducted
Types of bioassays: Chesapeake Biological Laboratory has three investigators who do various kinds of cellular
biochemical toxicology using aquatic organisms. They are equipped and capable of doing a variety of sublethal
biomarker-type assays on Chesapeake Bay organisms. They are capable of doing many others as needs or
opportunity allow. (See tables for description of exact testing capabilities.)
Future plans: CBL plans to investigate solid phase sediment bioassays (also pore water, etc.); factors related to
equilibrium partitioning; and adaptation to larval fish.
Field sampling capabilities: full range, including research vessels
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: some work in electron microscopy, spectrometry, and chromatography
Major equipment available:
electron microscopy: SEM
spectrometry: 2 GC/MS
alpha, beta, gamma counting
2AA's
UV-VIS
fluorescence
chromatography: GC
HPLC
TLC
CC
1C, etc.
Additional Resources
Computerized information retrieval services available: full range of PC's; Macintoshes; DEC-VAX; library with full
search capabilities
v Computers.and.software available: full complement of contemporary softwareachromatography data systems, etc.
Quality Assurance/Quality Control Procedures and Capabilities
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QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FTFRA of August 17, 1989?
Not necessarily; CBL is a typical academic research laboratory doing research which is not used for regulatory
purposes.
Follow study protocols? generally
Have a complete set of Standard Operating Procedures? some do; some do not
Archive facility for the data generated? Yes
Computer-generated data? Yes
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Coastal Bioanalysts, Inc.
Peter F. De Lisle or Ruth L. Williams Testing site: Coastal Bioanalysts, Inc.
P.O. Box 626 Gloucester Point Office Plaza,
Gloucester Point, Virginia 23062 Unit C
804-642-0168 Route 17
Gloucester Point, Virginia 23062
Hours: 8:00 AM - 4:30 PM
daily
Bioassays Conducted
Types of bioassays: Coastal Bioanalysts, Inc. (CBI) conducts acute and chronic bioassays with both fish and
invertebrates. (See tables for descriptions of exact test capabilities.)
Other bioassay work: CBI also performs field impact studies in both freshwater and marine environments for
benthos, plankton, and nekton. Ambient toxicity is addressed. Toxicity Reduction Evaluations, both municipal and
industrial, are performed. The lab also develops site-specific water quality criteria/standards.
Future plans: CBI expects to increase the size of its physical plant within the next two years. Modifications
necessary to permit flow-through/bioconcentration studies (freshwater & marine) are planned at that time. In
addition, increased capacity for static/static renewal tests will be available within the next year. If the need arises,
addition of a mobile lab for on-site testing is possible.
Field sampling capabilities:
Water: 4 ISCO composite samplers, 2 equipped for Priority Pollutants (at Bionetics)
Sediment: coring devices, samplers: Ponar, Smith-Mac, Peterson, Ekman (leased)
Organisms: Surber sampler, seine net, plankton nets (leased)
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: Through a joint association with the Bionetics Corporation, CBI can perform
analyses for organics, inorganics and metals in tissue, water, wastewater, sediment and soil samples. Full GC/MS,
ICP, GC, AA, and TOX capability is provided.
Major equipment available:
electron microscopy: none
spectrophotometry: 1 VG Trio GC/MS with full library search & retrieval capability
1 Perkin-Elmer Model 3939 AA with graphite furnace
1 IL S12 AA with graphite furnace
1 JY 24 Sequential ICP with autosampler
CLP interface program for GC/MS and ICP
VG 4 station network data system for above
chromatography: 2 Model 3300 Varian GC
1 Model 3400 Varian GC with autosampler
1 Water GC with autosampler
3 Tekmar Purge and Trap devices
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Detectors: 2 FID, 4 BCD, 2 PID, 1 Hall, 1 FPD
CLP program interface for GC's
Additional Resources
Computerized information retrieval services available: Acquisition of public-access online data bases (e.g.
DIALOG) is possible.
Computers and software available: PC's, modem; EPA software for calculation of ECSO's, probit analysis, analysis
of variance and other statistical software; word processing, data management packages.
Staff available: full-time: 2 Ph.D.'s; 1 B.S.; 1 BA.
part-time: 1 non-degree
(See also Bionetics Corp.)
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FTFRA of August 17, 1989? Yes
In addition, this lab meets the requirements of NPDES biomonitoring program; e.g., reference toxicity tests are
performed on routine basis. Through a cooperative agreement with the Bionetics Corporation, CBI can also provide
state certified (MD, VA, NC) chemical analyses and GLP chemical support of TSCA/FIFRA work.
Follow study protocols? Yes* Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes
Computer-generated data? No**
*Protocols for effluent biomonitoring are approved by Virginia Water Control Board. All protocols submitted to date
in Maryland have also been approved.
**Computer systems of Bionetics Corp. (providing chemical support) are validated.
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Commonwealth Laboratory, Inc.
Edwin Cox III Testing site: Commonwealth Laboratory, Inc.
2209 E. Broad Street 305, 307 N. 26th Street
Richmond, Virginia 23223 Richmond, Virginia
804-648-8358 Hours: 8:30 AM - 5:00 PM
FAX 804-644-5820 7 days/week
Bioassays Conducted
Types of bioassays: Commonwealth Laboratory, Inc. (CLI) conducts acute and toxic bioassays of dapnia and fathead
minnows. (See tables for description of exact test capabilities.)
Future plans: CLI hopes to conduct in situ toxicity testing in 1990.
Field sampling capabilities: flow and time proportional samplers; depth samplers; bottom samplers
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: water, wastewater, solids, air
Major equipment available:
electron microscopy: none
spectrophotometry: Extrel GC/MS
2 EL AA *s with furnace
chromatography: Tracer GC
also: TOC, TOX, IR, UV and all analyses required by USEPA except
radiological
NTTS-certified for bulk asbestos identification
Additional Resources
Computerized information retrieval services available: some
Computers and software available: 1 Epson; 3 KAYPRO 286i
Staff available: full-time: 1 M.Ch.E.; 7 B.S.'s
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989?
No; the lab is in accordance with USEPA Waste Water and Drinking Water, but is not specifically inspected for
TSCA and FIFRA.
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
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Do you have an archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems validated? No
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EA Engineering, Science, and Technology, Inc.
Wayne L. McCulloch Testing site: EA Engineering, Science, & Tech.
EA Mid-Atlantic Regional Operations 15 Loveton Circle
Hunt Valley/Loveton Center Sparks, Maryland 21152
15 Loveton Circle Hours: business: 8:30 AM - 5:00 PM
Sparks, Maryland 21152 Monday - Friday
301-771^950 operating: 8:00 AM - 7:00 PM
FAX 301-771-4204 daily
Bioassays Conducted
Types of bioassays: EA Engineering conducts acute and chronic bioassays on fish, invertebrates and plants. (See
tables for description of exact testing
capabilities.)
Other bioassay work: EA Engineering can perform Toxicity Identification Evaluations (TIE's) to identify the sources
and types of toxicants. They also use bioassays to assist in the development of design-treatability studies for
wastewater treatment plants (WWTP's), and can provide consulting services, regulatory interactions and permit
review.
Future plans: EA Engineering plans to increase capabilities to perform more TIE/TRES studies, to develop better
GLP facilities to provide such services to clients requiring these tests, and to increase consulting facilities.
Field sampling capabilities:
Water: McNeils, Mannings & Iscus composite samplers; metering pumps; Van Dorns or Nishen bottles (for samples
at various depths); trash pumps (for large volume water sampling)
Sediment and organisms: Ponar, Ekman; Hess samplers; various nets, seines, and trawls; electroshocking; boats and
prams
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: standard priority pollutants; hazardous waste; GC/MS; HPLC's; CLP analyses
Major equipment available:
electron microscopy: none
spectrophotometry: 6 GC/MS
4 Tekmar Purge and Trap devices
1 ICP/AA spectrophotometer
1 AA spec.
2 UV/VIS spec's
chromatography: 5 GLC
various detectors
1 HPLC
1 GPC
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Additional Resources
Computerized information retrieval services; available: DIALOG, National Library of Medicine, Dialcom, Chemical
Information Systems, Storit
Computers and software available: mainframe VAX and mini-VAX; 5 IBM PC's; 8 laser printers; Perkin-Elmer
hardware (LIMS, CLAS); Word II; Word Perfect; SAS; Lotus 1-2-3; Symphony; dBase; Toxstat; Toxcalc; EPA acute
and chronic calculation programs, etc.
Staff available: 4 PhJD.'s; 5 M.S.'s; 1 M.E.M.; 2 BA.'s
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and F1FRA of August 17, 1989? Yes
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems validated? Yes
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Environmental Laboratories, Inc.
Steven Pond Testing site: Environmental Laboratories, Inc.
9211 Burge Ave. 9211 Burge Ave.
Richmond, Virginia 23237 Richmond, Virginia 23237
804-271-3440 Hours 24 hrs./day
FAX 804-271-1313
Bioassays Conducted
Types of bioassays: Environmental Laboratories, Inc. does not conduct bioassays; they provide chemical analysis
of environmental samples only.
Future plans: analytical chemistry only
Field sampling capabilities: two full time experienced field services personnel; equipment and personnel for
sampling water wastewater, soils, sediments
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: chemical analysis of soils, sediments, water and wastewater for inorganics,
organics, trace metals, etc.
Major equipment available:
electron microscopy: none; but do have polarized light microscopy and plain light microscopy with phase
contrast
spectrometry: 1 Perkin-Elmer 2830 AA with furnace
1 Perkin-Elmer 5000 AA with furnace
2 each Hewlett Packard 5890 GC's with 5970 MS
1 Perkin-Elmer 8500 with HALC and PID detectors
chromatography: 2 Hewlett Packard GC's: 5830, 5710
1 Waters HPLC
Additional Resources
Computerized information retrieval services available: Laboratory Information Management System (LIMS); radian
SAM program which tracks samples, accumulates QC and sample data
Computers and software available: 15 IBM PC XT and AT computers; Compaq 286; and Zenith portable with 20
meg hard drive
Staff available: - 10 chemists
- 6 biologists
- 1 microbiologist
- 5 support
Quality Assurance/Quality Control Procedures and Capabilities
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QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? Yes
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? not reported
Archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems validated? Yes
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Environmental Resources Management, Inc.
Robert L. Dwyer, Ph.D. Testing site: ERM, Inc.
116 Defense Highway 855 Springdale Drive
Suite 300 Exton, Pennsylvania 19341
Annapolis, Maryland 21401 Hours: 8:00 AM - 5:00 PM
301-266-0006 Monday - Friday
FAX 301-266-8912 additionally by appointment
Bioassays Conducted
Types of bioassays: Environmental Resources Management, Inc. (ERM) conducts acute and chronic bioassays with
both fish and invertebrates. They also conduct algal growth studies and Microtox assays.
Other bioassay work: ERM can conduct on-site bioassays (setting up labs at clients' facilities) and ambient toxicity
assays (caging sutdies).
Future plans: ERM will be gearing up to do bioassay testing as it becomes necessary for NPDES DMR monitoring.
They will also focus on updates of TIE/TRE protocols released by EPA.
Field sampling capabilities:
Watensmall boat sampling capabilities; stream sampling capabilities
Benthos and sediment: Ponar grab, diver-operated cores
Fish: seines, gill nets and electroshock equipment
Plankton: bongo net
(All Chesapeake Bay sampling is coordinated and staged from Annapolis office; biology lab in Exton is equipped
and staffed for all taxonomic identification.)
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: toxics in the aqueous and solid media; support of activities under CERCLA,
RCRA, WQA of 1987, as well as state regulations; industrial hygiene and occupational health analyses
Major equipment available:
electron microscopy: none at present; planning to add TEM/SEM in 1990
spectrophotometry: flame and furnace AA's with ICP
total organic carbon analysis
visible wavelength spectrophotometer
Fourier transformed infrared analyzer
chromatography: 3 GC's
2GC/MS's
total halogen analyzer
Additional Resources
Computerized information retrieval services available: complete library retrieval services available (e.g. DIALOG,
BIOSIS); access to special databases (CHEMFATE) and government databases (USGS, WATERNET, NAWDEX,
EPA-REACH, GEMS)
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Computers and software available: Macintosh SB's and IPs linked using Applenet. All have Excel, word pro-
cessors, and MacDraw. 286/386 PC's for statistics, large DBMS. Modeling capabilities include most EPA-approved
packages (ERL Athens Center for Exposure Assessment Modeling, International Ground Water Modeling Center,
UNAMAP air models). Remote access to mainframe/mini's for SAS, large databases, etc.
Staff available:
Annapolis: 2 Ph.D.'s; 1 M.S.; 2 B.S.'s
Exton, PA: 2 M.S.'s; 1 BA.; 1 B.S. Also approximately 60 field professionals in these two locations.
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FTFRA of August 17, 1989?
No, neither lab has had an independent GLP audit. Managers believe themselves to be generally in compliance with
GLP and could set up easily to do tests for TSCA PMN's and FIFRA product
registrations.
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems validated? Yes
Additional explanation: Computer data storage/retrieval usually customized to meet needs of client. Systems can
be designed by subsidiary (ERM Computer Service, Inc.) to comply with CERCLA CLP requirements or TSCA
CBI data security protocols as needed.
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Environmental Systems Service, Ltd.
Dennis T. Brown, Lab Manager Testing site: Environmental Systems Service, Ltd.
218 N. Main St. 218 N. Main St.
Culpeper, Virginia 22701 Culpeper, Virginia 22701
703-825-6660 Hours: 8:00 AM - 5:00 PM
FAX 703-825-4961 Monday - Friday
(analysts Saturday and Sunday)
Bioassays Conducted
Types of bioassays: Environmental Systems Service, Ltd. conducts acute and chronic bioassays on fish, invertebrates
and algae. (See tables for description of exact testing capabilities.)
Field sampling capabilities: auto samplers, sediment grab samplers, macro-invertebrate bottom samplers
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: chemical analysis of water and wastewater; microbiological analysis of same;
food/dairy analysis. Certified in VA and MD.
Major equipment available:
electron microscopy: none
spectrophotometry: furnace and flame AA spectrophotometer
GC/MS
chromatography: GC
HPLC (1C)
TLC
Additional Resources
Computerized information retrieval services available: none reported
Computers and software available: IBM PC's; Wang system; in-house lab-tracking system; Lotus 1-2-3; dBase,
Paradox
Staff available: 4 M.S.; 5 B.S.; 1 A.S.; 1 high school
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? Yes
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes
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Computer-generated data? Yes Are these computer systems validated? Yes
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Experimental Pathology Laboratories, Inc.
Dr. Marilyn J. Wolfe Testing site: Experimental Pathology Lab., Inc.
P.O. Box 474 22866 Shaw Road
Herndon, Virginia 22070 Sterling, Virginia 22170
703-471-7060 Hours: 8:00 AM - 5:00 PM
FAX 703-471-8447 Monday - Friday
Bioassays Conducted
Types of bioassays: Experimental Pathology Laboratories, Inc. provides pathology services for invertebrate species
(e.g. various crustaceans and insects) and for finfish.
Futher description of bioassay work: EPL's services include necropsy of animals, preparation of tissues for
histologic sections, preparation of glass slides for light microscopy, preparation of tissues for electron microscopy
and pathologic evaluation of tissues. EPL has processed fish of various ages
(larvae through adults) and a variety of species including fathead minnows, bluegill sunfish, green sunfish, striped
bass, channel catfish, sheepshead minnows, shad, largemouth bass, Japanese medaka, guppies, goldfish and northern
pike. Invertebrate species that have been processed include grass shrimp and mayfly nymphs. Turnaround time from
receipt of tissues to a complete report is variable depending upon the size and number of specimens. Services
include consultation for the planning of toxicologic studies with aquatic species under controlled laboratory conditions
of surveys or aquatic species in natural waterways.
Future plans: EPL is expanding its expertise in invertebrate pathology to include various species of mollusks and
other crustaceans.
Field sampling capabilities: EPL will provide pathologists and/or technical staff to perform necropsies on animals
in the field and to preserve tissues for histopathology.
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: not reported
Major equipment available:
electron microscopy: Hitachi 12A transmission electron microscope is available for examination of animal
tissues.
spectrophotometry: none
chromatography: none
Additional Resources
Computerized information retrieval services available: none
Computers and software available: EPL has a computer system in which histopathology data generated by the
pathologist is stored and processed for reporting purposes. The computers are IBM-compatible.
Staff available: 1 Ph.D.; 1 M.S.
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Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
Is it in compliance with the Good Lab Practices requirements under the Toxic Substances Control Act and the
Federal Insecticide, Fungicide, and Rodenticide Act of August 17, 1989? Yes
Follow study protocols? Yes Have a complete set of. Standard Operating Procedures? Yes < v-*
Archive facility for the data generated? Yes (EPL also has a residual materials archive facility.)
Computer-generated data? Yes; data is generated by a pathologist and entered into a computer.*
Are these computer systems validated? Yes
"Occasionally a study that is done by EPL requires morphometry. Measurement data is computer-generated in that
the computer may calculate an area, e.g.,
from a perimeter outlined by a technician on a microscope slide or a photomicrograph.
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Free-Col Laboratories, Inc.
William R. Osman Testing site: Free-Col Laboratories, Inc.
P.O. Box 557 Cotton Road
Cotton Road Meadville, Pennsylvania 16335
Meadville, Pennsylvania 16335 Hours: 7:00 AM - 9:30 PM
814-724-6242 Monday - Friday
FAX 814-333-1466 (7 days/wk for some bioassay work)
Bioassays Conducted
Types of bioassays: Free-Col Laboratories conducts acute and chronic bioassays with both fish and invertebrates.
They also conduct Microtox bacterial toxicity tests. (See tables for more complete descriptions of bioassay
conditions.)
Future plans: EPA Test Method 1003.0 algal(Selenastmm capricornutum ) growth test
Field sampling capabilities: 3 ISCO autosamplers
3 American Sigma autosampler
3 Wildco Kemmerer samplers
1 Wildco bottom sampler
10 artificial substrates
2 Ekman dredges
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: Free-Col Laboratories, Inc. is a full service independent testing facility specializing
in analytical services for environmental protection, occupational health, industrial hygiene, food processing and
process quality control. Areas of focus are RCRA and Superfund analyses, technical services, industrial hygiene,
hazardous waste and groundwater monitoring, wastewater and drinking water analyses and toxicity (bioassay) testing
for clients nationwide and overseas.
Major equipment available:
electron microscopy: none reported
spectrophotometry: GC/MS: Finnigan Model 4000, Finnigan Model 5100, Hewlett-
Packard Model 597 MSB
AA: Perkin-Elmer Model 360, Perkin-Elmer Model 3030, Perkin-Elmer Model 5500,
Perkin-Elmer Model 5100/Zeeman
chromatography: GC: Perkin-Elmer Model Sigma 3B, Perkin-Elmer Model 8700, 2 Hewlett-Packard
Models 5880A
HPLC: Waters Model 510 with 430 heater
Additional Resources
Computerized information retrieval services available: none reported
Computers and software available: Perkin-Elmer LJMS/2000 System 3205 CPU; IBM PC's; EPA Dunnetts and
Probit; Toxstat from University of Wyoming; Symphony,
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WordPerfect
Staff available: not reported
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? Yes
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems validated? Yes
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Hampton Roads Sanitation District
George D. Kennedy Testing site: HRSD Special Projects Lab
P.O. Box 5000 1436 Air Rail Avenue
"Virginia Beach, Virginia 23455 Virginia Beach, Virginia 23455
804-460-2261
FAX 804-460-2372
Bioassays Conducted
Types of bioassays: Hampton Roads Sanitation District (HRSD) conducts acute and chronic bioassays on estuarine
fish and invertebrates. (See tables for exact testing capabilities.)
Other bioassay work: HRSD currently conducts NPDES required tests for 3 POTW's. Quarterly testing require-
ments on each include at least 2 acute and 1 chronic test.
Future plans: HRSD would like to conduct NPDES required tests for 9 POTW's.
Field sampling capabilities: 25' fully-equipped oceanographic research vessel
Loran navigational recording fathometer
submersible pump assembly or NIO bottles
otter trawls, hand seines, and traps
Ponar and Van Veen sediment grabs
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: wastewater treatment process control; industrial waste characterization; metals;
nutrients
Major equipment available:
electron microscopy: none
spectrophotometry: AA spectrophotometer
UV/VES spectroscopy
auto analyzer colorimetric system
Additional Resources
Computers and software available: IBM-compatibles: HP ES12, Compaq 286's; Macintosh SB's;
Lotus 1-2-3; WordPerfect 5.0; Revelation; Statgraphics; SAS; Harvard Graphics; Excel
Staff available: toxicity: 2 Ph.D's; 3 B.S.'s; 3 A.A.'s
chemistry: 1 Ph.D.; 6 M.S.'s; 2 B.S. or equivalent
statistics/computer: 2 M.S.'s
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
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In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? Yes
Representatives from HRSD have not read FIFRA and TSCA GLP of August 17,1989. Their QA/QC program was
developed based on the
EPA acute and chronic manual and previous GLP's.
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes
Computer-generated data? *
*HRSD has EPA programs to calculate LC's and NOEC's. They do not store data or transfer original lab work
sheets onto the computer.
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Horn Point Environmental Laboratory
University of Maryland, Center for
Environmental and Estuarine Studies Testing site: Horn Point Environmental Lab
Ian HartwelV Charles Hocutt UMCEES
P.O. Box 775 P.O. Box 775
Cambridge, Maryland 21613 Cambridge, Maryland 21613
301-228-8200 Hours: not reported
FAX 301-476-5490
Bioassays Conducted
Types of bioassays: Horn Point Environmental Lab conducts extensive acute and chronic bioassays on fish,
invertebrates, and various algae, for all salinity ranges. (See tables for exact test capabilities.)
Future plans: none beyond those described
Field sampling capabilities: seines, trawls, trap nets, dip nets, etc.; ISCO water samplers; bottom dredge
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: effluents — basic water chemistry and autoanalyser
Major equipment available:
electron microscopy: none
spectrophotometry: flame AA spec.
4MS's
chromatography: 3 GC's
1C
2HPLC's
detectors: TCD, 2 FID, EC
(Above equipment is primarily dedicated to nutrient analysis.)
Additional Resources
Computerized information retrieval services available: DIALOG
Computers and software available: VAX 780; VAX 750; Micro VAX; IBM PC's; access to supercomputing
Staff available: 2 Ph.D.'s; 2 M.S. students
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? No
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? No
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Archive facility for the data generated? No
Computer-generated data? No
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James R. Reed and Associates
Elaine Glover or Liz Christoff Testing site: James R. Reed and Associates
813 Forrest Drive 813 Forrest Drive
Newport News, "Virginia 23606 Newport News, Virginia 23606
804-599-6750 Hours: 8:00 AM - 5:00 PM
FAX 804-591-7680 Monday - Friday
Bioassays Conducted
Types of bioassays: James R. Reed and Associates conducts acute and chronic bioassays with both fish and
invertebrates. They also perform algal growth tests. (See tables for exact test capabilities.)
Other bioassay work: In addition to routine bioassay work, this facility is experienced in conducting site-specific
studies and developing protocols for
indigenous (non-standard) test organisms.
Future plans: James R. Reed and Associates plans to begin conducting bioassays in association with Toxicity
Identification Evaluations and Toxicity Reduction Evaluations for NPDES dischargers within the next year. Within
the next three years, the lab plans to develop lexicological capabilities for sediment analysis and begin operation of
a mobile laboratory unit.
Field sampling capabilities:
Water: grab; 24-hr, composite and/or 7-day composite water sampling with Isco pump; groundwater monitoring using
bailers or dedicated tubing; subsurface water collection using Isco pump and BETA plus horizontal water sample
bottles
Sediment: Ponar grab; core sampling; dredging; extraction of water from soil using a lysimeter
Macrobenthic sampling: D-frame nets
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty:
wastewater/NPDES dischargers (priority/non-priority pollutants)
groundwater monitoring
drinking water scans
soils analysis
landfill leachates
Major equipment available:
electron microscopy: none
spectrophotometry: Finnigan GC/MS
Varian AA spec. (flame and graphite furnace)
Beckman spectrophotometer
chromatography: Varian GC
BCD, FID detectors
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Additional Resources
Computerized information retrieval services available: lexicological Network (TOXNET) available 24-hr.
Computers and software available: 6 IBM PC-compatible computers with Multimate word processor, Quatro
spreadsheet and graphics and Toxistat/EPA statistical programs
Staff available: 1 Ph.D; 2 M.S.'s; 12 B.S.'s
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989?
This lab is in compliance with GLP standards under the Toxics Substances Control Act of November 29,1983 with
the following exception: due to the small size of the laboratory, it has no in-h
ouse chain of custody documentation other than a labsheet which has all required information.
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems validated? No
Additional explanation: Computer printout is validated for computer-generated data, and hand calculations are
available in SOP's.
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Johns Hopkins University
Applied Physics Lab Testing site: Johns Hopkins University
Dennis T. Burton Applied Physics Laboratory
4800 Atwell Road 4800 Atwell Road
Shady Side, Maryland 20764 Shady Side, Maryland 20764
301-867-7000 Hours: 8:30 AM - 5:00 PM
FAX 301-867-0839 Monday - Friday
Variable hours Saturday and Sunday
Bioassays Conducted
Types of bioassays: The JHU/APL Toxicology and Bioassay Facility can conduct the following types of aquatic
bioassays: acute, short-term chronic, partial life stage, early life stage (ELS) and full life cycle tests. (See the
Applied Physics Lab row in the attached tables for more specific information about these bioassays.)
Other bioassay work: Several estuarine and freshwater fish, invertebrates, and algae are maintained in culture at
the facility. The laboratory can perform toxicity tests with contaminants in water, sediments, and groundwater, as
well as media with toxic volatile compounds. JHU/APL performs all of the acute and short-term chronic tests for
the Maryland Department of the Environment NPDES permits compliance division, and has produced two manuals:
"Standard Operating Procedures for Acute Effluent Toxicity Tests with Freshwater and Saltwater Organisms" and
"Standard Operating Procedures for Short-Term Chronic Effluent Toxicity Tests with Freshwater and Saltwater
Organisms."
Future plans: The JHU/APL Toxicological and Bioassay Facility located in Shady Side, Maryland, will be moving
during the summer of 1990 from JHU/APL to the University of Maryland Agricultural Experiment Station Wye
Research and Education Center, Queenstown, Maryland. The move will increase our present toxicity testing space
from 2,000 to 3,000* sq. ft. Chemistry laboratory space will be increased from 900 sq. ft. to -1,600 sq. ft. at the
Wye Center.
field sampling capabilities:
Water: Niskin bottles (single or Rosette), Kemmerer samplers, pumping, etc.; rotating disk and flat plate samplers
are used to sample surface microlayers.
Organisms: boats, seines, push nets, plankton nets, traps, artificial substrates, Hess samplers, Surber samplers and
other devices
Sediments: single and multiple core samplers, Ekman dredges, Kellen grab, Ponar grab, etc.
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: heavy metals, several classes of organics, nutrients
Major equipment available:
electron microscopy: none
spectrophotometry: Perkin-Elmer AA Model 2380 equipped with an HGA 400 graphite furnace and
MHZ-200 mercury-hydride system
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chromatography: Waters Associates HPLC Model 680 controller equipped with a Model 481 variable
wavelength UV spectrophotometer
Model 740 data integrator, and Model 712 WISP automatic sampler
Hewlett Packard GC Model 5890 equipped with flame ionization and flame
photometric detectors and an HP 3393 A integrator and HP 7673 automatic sampler
(An ffiM-AT PC with Interactive Microware software is used for HPLC and GC
control and sample analysis.)
Additional Resources
Computerized information retrieval services available:
- Access via PC modem to the DIALOG Inc., the Orbit System, BRS, and DOE-RECON Literature Search.
- DIALOG and Orbit Systems provide access to over 350 other data bases which include the NTIS and Dissertation
Abstracts.
- Other data bases that can be accessed are BIOS Previews from Biological Abstracts Inc., Chemical Abstracts
Service, Oceanic Abstracts, Pollution Abstracts, Environline, EMBASE, Water Resources Abstracts, Aquatic Science
and Fisheries Abstracts, etc.
Computers and software available:
- several IBM AT PC's
- WordPerfect 5.0, Symphony 2.0, Lotus 1-2-3, Sigmaplot 3.1, Norton's Utility Microsoft Chart 3.0, PC SAS, Toxstat
2.0, EPA Bioassay Statistical Program, etc.
- access via modems to the IBM mainframe computers at the Applied Physics Laboratory which contain many
software programs, programming languages, and graphic art capabilities
Staff available: 3 Ph.D.'s; 1 M.A.; 2 M.S.'s; 3 BA.'s; 3 B.A-'s
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and F1FRA of August 17, 1989? Yes
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems validated? Yes
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Malcolm Pirnie, Inc.
Jane Hughes or Meryl Alexander Testing site: 100 Grassland Road
100 Grasslands Road Elmsford, New York 10523
Elmsford, New York 10523 (also) 301 Hiden Blvd.
914.347.2974 P.O. Box 6129
FAX 914-347-2984 Newport News, Virginia 23606
Hours: Monday - Friday
8:00 AM - 5:00 PM
Bioassays Conducted
Types of bioassays: Malcolm Pirnie conducts acute and chronic bioassays on fish and invertebrates. (See tables for
exact test capabilities.)
Future plans: Malcolm Pirnie may begin microcosm studies.
Field sampling capabilities: Ponar grab sampler, sediment corers, water samplers (Van Dorn, etc.), electroshocker,
swines, plankton nets, periphyton samplers
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: analyses of water, wastewater, hazardous waste, sediments, sludges
Major equipment available:
electron microscopy: none
spectrophotometry: 1 MS
1 AA spec.
2 spectrophotometers
chromatography: 1 GC/FID
1 HPLC/UV detector
1 GC/ECD
1 GC/NPD
1 GC/MS
1 Purge and Trap device
Additional Resources
Computerized information retrieval services available: DIALOG - includes Aquatic Science and Fisheries Abstracts,
BIOSIS, Enviroline, etc.; CIS - includes OHM/TADS, AQUIRE, ENVTROFATE, CHRIS, CESARS, PHYTOTOX,
etc.
Computers and software available: LIMS is being implemented for lab; Prime minicomputer with assorted
engineering software, including CAD; > 200 personal computers (IBM-compatible) with assorted software for word
processing (WordPerfect, PC Write) for spreadsheets (1-2-3) and statistics (SAS, Statgraphics); desktop publishing
and graphics
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Staff available: full-time: (chemistry) 2 M.S.'s; 3 B.S.'s; 2 A.S.'s
part-time: (toxicology) 1 M.S.; 1 M.S.P.H
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? Yes
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems validated? *
*Computer programs are run with standard data sets; programs and outputs arc then part of SOP's.
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Olver Incorporated
Robert M. Roberts, RE. Testing site: Olver Incorporated
1116 South Main Street 1116 South Main Street
Blacksburg, Virginia 24060 Blacksburg, Virginia 24060
703-552-5548 Hours: 8:00 AM - 5:00 PM
FAX 703-552-5577 Monday - Friday
Bioassays Conducted
Types of bioassays: Olver Incorporated conducts acute and chronic bioassays using both fish and invertebrates.
They also perform chronic algal growth tests. (See tables for descriptions of exact test capabilities. Note that the
"number of tests our lab can run simultaneously" is based on running one test type only.)
Other bioassay work: Olver Incorporated has the capability and capacity to provide screening and definitive tests
using many different species, such as Lepomis machrochirus, Salmo gairdneri, Ictalurus punctatus, and others, simply
by obtaining these species from commercial suppliers. Chronic survival and reproduction testing is also possible for
D. pulex and D. magna, although methods are not presented for these organisms in the EPA manual EPA/600/4-
89/001, March 1989.
Future plans: Facilities are being expanded in order to have more capacity for the same types of testing currently
conducted. Within six months they hope to double the number of tests they can perform simultaneously. Olver also
plans to expand its capabilities to accommodate Toxicity Reduction Evaluations as well as site-specific studies. They
will continue trial seawater tests, with the intent of offering these commercially at some time.
Field sampling capabilities: Olver Incorporated has a full range of field sampling capabilities, including qualified
technicians and equipment. Routine collections of water, wastewater, sediment, sludge, soil and hazardous
materials/waste samples are made. The laboratory staff also perform macroinvertebrate stream surveys and collect
native test organisms.
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: inorganic chemistry; wet chemistry; microbiological tests; treatability studies;
Toxicity Reduction Evaluations; EP Toxicity and TCLP extractions
Major equipment available:
electron microscopy: none
spectrophotometry: 1 Perkin-Elmer 1100 AA spectrophotometer
1 Perkin-Elmer 305B AA spectrophotometer
chromatography: 1 HP 5730A GC
1 HP 5780A GC
Additional Resources
Computerized information retrieval services available: none
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Computers and software available: IBM System 3600; IBM PC's; Compaq 386; EPA statistical programs for the
analysis of lexicological data (LC 50 determination); Toxstat statistical programs for the analysis of lexicological
data
Staff available: full-time: 1 M.S.; 2 B.S.'s
part-time: 1 B.S.
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of Augusl 17, 1989? Yes
(This lab is in the process of complying with the Good Lab Practices requirements of TSCA and FIFRA.)
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes
Computer-generated dala? Yes Are ihese computer systems validated? Yes
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Patuxent Wildlife Research Center
U.S. Fish and Wildlife Service Testing site: Patuxent Wildlife Research Center
Dr. Harry N. Coulombe U.S. Fish and Wildlife Service
Laurel, Maryland 20708 Laurel, Maryland 20708
301-498-0279 Hours: 8:00 AM - 4:30 PM
FAX 301-497-0515 Monday - Friday
Bioassays Conducted
Types of bioassays: The Patuxent Wildlife Research Center conducts autotrophic and microcosm system tests and
photosynthesis tests on the sago pondweed, Potamogeton pectinatus.
Further description of bioassay work: The above test systems are in various stages of development. The Wildlife
Research Center does not run standard analyses, except for several herbicides used to develop the systems. The three
laboratory procedures need to be verified by in situ tests (i.e., floating pods which are currently being tested). The
lab systems use synthetic or natural medias and substrates and can be used to test the effects on plant growth and
morphology of single chemicals, combinations of chemicals, and other environmental stressors (e.g., light intensity).
Large numbers of explants can be propagated (cloned) using axenic techniques.
Future plans: The Patuxent Wildlife Research Center plans to verify tests for the three laboratory procedures as
described above and to utilize procedures
in large scale testing.
Field sampling capabilities: two boats: 17* whaler and 21' Roballo; box core sampler, benthic air-lift device;
Solomat water quality analyzer
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: analysis of tissue, soil, water, and plant samples for pesticides, metals, and PAH's
Major equipment available:
electron microscopy: none
spectrophotometry: MS: 2 routine analysis mass specific detector GC/MS systems (HP); 1 research-grade
GC-MS/MS system (Finnigan)
AA: 3 Perkin-Elmer graphite furnace systems; 2 flame AA systems and 2 cold vapor
mercury analyzers
ICP: 1 Perkin-Elmer ICP Arc. system for analyses of inorganic chemicals
chromatography: GC's: several GC's; packed systems (3) and capillary systems (10) with computer
aided data capture (Nelson) and various detectors (PID, FPD, NPD, BCD, and FID)
LG: multiple HPLC capabilities with various detectors, UV, VIS, both variable and
fixed wavelength, and continuously variable fluorometric conductivity detectors
Maxima automated data handling station
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Additional Resources
Computerized information retrieval services available: Library has on-line access to two literature data bases —
DIALOG and OCLC.
Computers and software available: IBM-compatible and Apple PC's; Hewlett Packard 3000 minicomputer; Prime
9955-11 minicomputer, access via terminals and phone lines to NIH computing facility; variety of statistical,
spreadsheet, graphics and word processing packages; programmers also available.
Staff available: 19 Ph.D.'s; 12 B.S.'s; 6 M.S.'s
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
Is it in compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? No
Patuxent's work does not usually involve FIFRA. Consequently, they have not adopted all of the GLP procedures.
They feel that the quality of the data they generate is as good or better than that of any facility in full compliance.
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? No*
Archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems validated? No
*SOP's are being developed.
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Riverside Laboratories
Gloria Gibson Testing site: Riverside Laboratories
1300 Old Denbigh Boulevard 1300 Old Denbigh Boulevard
Newport News, Virginia 23602 Newport News, Virginia 23602
804-886-3900 Hours: 24 hours a day
FAX 804-886-3988
Bioassays Conducted
Types of bioassays: Riverside Laboratories does not report capabilities for standard bioassay work. They do conduct
histological and microbiological studies.
Other bioassay work: Riverside Laboratories conducts work on a full range of organisms pathogenic to man.
Future plans: DNA probes for pathogenic organisms.
Field sampling capabilities: none
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: complete analytical biochemistry testing on tissue, blood, urine
Major equipment available:
electron microscopy: SEM
spectrophotometry: GC/MS
Bio Chemical Analyzer
AA with graphite furnace and cold
Radio Isotopes Gamma and Beta counter
ICP
chromatography: GC
Additional Resources
Computerized information retrieval services available: Yes
Computers and software available: Reporting and statistical work with Cerner system.
Staff available: not reported
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? Yes
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
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Archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems vah'dated? Yes
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Technical Testing Laboratories
Mr. Dilip V. Kalyani, P.E. Testing site: Technical Testing Laboratories
4643 Benson Avenue 4643 Benson Avenue
Baltimore, Maryland 21227 Baltimore, Maryland 21227
301-247-7400 Hours: 8 hrs./day
FAX 301-247-7402 7 days/week
Bioassays Conducted
Types of bioassays: Technical Testing Laboratories conducts acute and chronic bioassays with both fish and
invertebrates. (See tables for descriptions of
exact test capabilities.)
Other bioassay work: Technical Testing Laboratories is also equipped to perform Microtox toxicity tests.
Future plans: Extensive Toxicity Reduction Evaluations (TRE) and Toxicity Identification Evaluations (TIE).
Field sampling capabilities: grab; 24-hour time proportional; 24-hour flow proportional; sediment sampling
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: inorganic non-metals; inorganic metals with AA, ICP, etc.; air; soil
Major equipment available:
electron microscopy: none
spectrophotometry: GC/MS, HPLC, TCLP, AA, ICP, TOX, TOC, FTIR, IR, etc.
chromatography: GC
Additional Resources
Computerized information retrieval services available: LIMS
Computers and software available: IBM and IBM-compatibles
Staff available (degree status, number, and time): full-time: 2 Ph.D.'s; 2 M.S.'s; 14 B.S.'s; 9 technicians
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? Yes
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems validated? Yes
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University of Maryland
Agricultural Experiment Station Testing site: University of Maryland
Lenwood Hall Agricultural Experiment Station
Wye Research and Education Center Wye Research and Education Center
P.O. Box 169 Queenstown, Maryland 21658
Queenstown, Maryland 21658
301-827-6202
FAX 301-827-9039
Note: This facility was undergoing reorganization at press time. For most of calendar year 1990, two stations will
be maintained - the previously established Johns Hopkins University Applied Physics lab (see entry by that title)
and a new facility developed at the Wye Research and Education Center named above. The capabilities for each
lab are listed as identical: contact either laboratory for more information.
Bioassays Conducted
Types of bioassays: The UM Agricultural Experiment Station can conduct the following types of aquatic bioassays:
acute, short-term chronic, partial life stage, early life stage (ELS) and full life cycle tests. (See the Applied Physics
Lab row of the tables in back for more specific bioassay descriptions.)
Other bioassay work: Several estuarine and freshwater fish, invertebrates, and algae are maintained in culture at
the facility. The laboratory can perform toxicity tests with contaminants in water, sediments, and groundwater, as
well as media with toxic volatile compounds.
Future plans: The JHU/APL Toxicological and Bioassay Facility located in Shady Side, Maryland, will be moving
during the summer of 1990 from JHU/APL to the University of Maryland Agricultural Experiment Station Wye
Research and Education Center, Queenstown, Maryland. The move will increase our present toxicity testing space
from 2,000 to 3,000* sq. ft. Chemistry laboratory space will be increased from 900 sq. ft. to -1,600 sq. ft. at the
Wye Center.
Field sampling capabilities:
Water: Niskin bottles (single or Rosette), Kemmerer samplers, pumping, etc.; rotating disk and flat plate samplers
are used to sample surface microlayers.
Organisms: boats, seines, push nets, plankton nets, traps, artificial substrates, Hess samplers, Surber samplers and
other devices
Sediments: single and multiple core samplers, Ekman dredges, Kellen grab, Ponar grab, etc.
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: heavy metals, several classes of organics, nutrients
Major equipment available:
electron microscopy: none
spectrophotometry: Perkin-Elmer AA Model 2380 equipped with an HGA 400 graphite furnace and MHZ
200 mercury-hydride system
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chromatography: Waters Associates HPLC Model 680 controller equipped with a Model 481 variable
wavelength UV spectrophotometer
Model 740 data integrator, and Model 712 WISP automatic sampler
Hewlett Packard GC Model 5890 equipped with flame ionization and flame
photometric detectors and an HP 3393 A integrator and HP 7673 automatic sampler
(An IBM-AT PC with Interactive Microware software is used for HPLC and GC
control and sample analysis.)
Additional Resources
Computerized information retrieval services available:
- Access via PC modem to the DIALOG Inc., the Orbit System, BRS, and DOE-RECON Literature Search.
- DIALOG and Orbit Systems provide access to over 350 other data bases which include the NTIS and Dissertation
Abstracts.
- Other data bases that can be accessed are BIOS Previews from Biological Abstracts Inc., Chemical Abstracts
Service, Oceanic Abstracts, Pollution Abstracts, Environline, EMBASE, Water Resources Abstracts, Aquatic Science
and Fisheries Abstracts, etc.
Computers and software available:
- several IBM AT PC's
- WordPerfect 5.0, Symphony 2.0, Lotus 1-2-3, Sigmaplot 3.1, Norton's Utility Microsoft Chart 3.0, PC SAS, Toxstat
2.0, EPA Bioassay Statistical Program, etc.
- access via modems to the IBM mainframe computers at the Applied Physics Laboratory which contain many
software programs, programming languages, and graphic art capabilities
Staff available: 3 Ph.D.'s; 1 M.A.; 2 M.S.'s; 3 BA.'s; 3 B.A.'s
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
-InwMnpliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? Yes
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes;
Computer-generated data? Yes Are these computer systems validated? Yes
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University of Maryland at Baltimore
Aquatic Toxicology Facility
Andrew S. Kane and R. Reimschuessel Testing site: UM School of Medicine
UM School of Medicine Department of Pathology --
Department of Pathology Aquatic Toxicology Facility
10 South Pine Street 610 West Lombard Street
Baltimore, Maryland 21201 . Baltimore, Maryland 21201
(301) 328-7230/7276 Hours: 24 hours per day
FAX (301) 328-8414 7 days per week
Bioassays Conducted
Types of bioassays: The Aquatic Toxicology Facility conducts both acute and chronic bioassays on fish. (See tables
for descriptions of exact test capabilities.)
Other bioassay work: Several flow-through systems are available for bioassay testing and are set up on a demand
basis. These include three solenoid-operated EnvironTox dilutors, three all-glass flow-through dosing systems with
pH and temperature control, and a 600-gallon polypropylene flow-through dilutor system. There is also an isolated
testing room for static or flow through testing. Additional on-site support facilities include histology and electron
microscopy laboratories, tissue and organ culture facilities, a diagnostic microbiology laboratory, office space and
a library.
Future plans: This facility is planning for expanded necropsy and tissue culture space; two additional isolated testing
rooms; and additional office space and a conference room.
Field sampling capabilities: Sampling and collection services are available and are arranged on a per-contract basis.
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: descriptive toxicology; diagnostic and comparative pathology; carcinogenesis assays
- in vivo and in vitro
Major equipment available:
electron microscopy: 2 JEOL 100B TEM's
1 AMR 1000 SEM
1 JEOL100CX TEM CAN with an ASID attachment used for transmission, scanning
transmission and secondary scanning electron microscopy. (All necessary equipment
for specimen preparation is present, including a polaron critical point drying apparatus
and a Techniques Hummer sputtering device for coating.)
spectrophotometry: Gilford "Response" automated spectrophotometer
Farrand ratio fluorometers
Perkin-Elmer MPF spectrofluorometer
Abbot TDX Polarized Fluorescence Scanner
chromatography: Waters HPLC system with two pumps and variable wavelength detector
Varian AA 575 spec.
145
-------
Varian 2700 GC
other: Corning Model 175 blood-gas analyzer
Beckman Astra 8, Kodak Ektachem 400 and Ektachrome 700 analyzers (for
electrolytes, urea nitrogen, protein, creatinine, enzymes, etc.)
complete facilities for enzyme and immuno-histochemistry
Additional Resources
Computerized information retrieval services available: Toxline, Medline,.MaryMed, BioAbstracts,. Zoological
Abstracts, Chemical Abstracts, Current Contents
Computers and software available: IBM ATs, Macintosh IPs, UMAB Mainframe; variety of word processing and
graphics packages, SAS, Lotus, Minitab, Montage
Staff available:
full-time: 1 V.M.D./Ph.D.; 1 Ph.D.; and 1 M.S.
part-time: 3 Ph.D.'s;! B.A./V.T; and 3 B.A.'s (some with M.S.)
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? Yes
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes;
Computer-generated data? Yes Are these computer systems validated? Yes, SAS
146
-------
University of Maryland, Baltimore County
Brian Bradley Testing site: UMBC
Department of Biological Sciences Dept. of Biological Sciences
Baltimore, Maryland 21228 Baltimore, Maryland 21228
301-455-2244 Hours: not reported
FAX 301-455-3875
Bioassays Conducted
Types of bioassays: UMBC does not conduct traditional bioassays.
Future plans: UMBC is aiming to have a rapid field test (based on stress-related protein synthesis) available in the
next 1 to 3 years. They also expect to have immuno diagnostic capability to identify specific contaminants
(or classes) in mixtures.
Field sampling capabilities: not reported
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: molecular/cellular biology
Major equipment available:
electron microscopy: several TEM
several SEM
spectrophotometry: full range: national center for MS
chromatography: full range
Additional Resources
Computerized information retrieval services available: BIOSIS, Medline, Environline, etc.
Computers and software available: MAC; VAX mainframe; IBM PC's
Staff available: 26 Ph.D.'s in department; one lab with 2 Ph.D.'s and 6 graduate students doing bioassays
(biochemical)
Quality Assurance/Quality Control Procedures and Capabilities
QA/QC program? No
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989?
No, but Radiation Safety Committee and Human Hazard Committee oversee all labs.
Follow study protocol? Yes Have a complete set of Standard Operating Procedures? not reported
Archive facility for the data generated? No
147
-------
Computer-generated data? No
148
-------
Versar, Inc., ESM Operations
Dr. Steve Weisberg Testing site: Versar, Inc., ESM Operations
9200 Rumsey Road 9200 Rumsey Road
Columbia, Maryland 21045 Columbia, Maryland 21045
301-964-9200 Hours: 8:00 AM - 4:30 PM
FAX 301-964-5156
Bioassays Conducted
Types of bioassays: Versar conducts acute and chronic bioassays using both fish and invertebrates. They also
conduct sediment bioassays andin situ bioassays with juvenile fish. (See tables for descriptions of exact test
capabilites.)
Future plans: Versar is currently renovating facility to enlarge holding and testing capabilities.
Field sampling capabilities: Five sampling boats equipped with a complete array of nets, trawls, sleds, corers, grabs,
and other sampling equipment. Back-pack and boat-mounted electroshockers, eel pots, minnow traps, crab pots, and
seines are also on-hand.
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: Versar's Springfield laboratory is a full service facility with capability to perform
organic and inorganic analyses on
priority pollutants in water, sediment, and biota.
Major equipment available:
electron microscopy: none
spectrophotometry: GC/MS
AA
ICP
UV-VIS scanning spectrophotometer
chromatography: GC
Ion chromatographs
HPLC
Additional Resources
Computerized information retrieval services available: DIALOG, MEDLARS, CIS systems for retrieval of ecological
and lexicological data and literature
Computers and software available: VAX/VMS operating system; PC's - Lotus, WordPerfect, Graphwriter, SAS,
EPA statistical programs for aquatic toxicology; Hewlett Packard plotters
Staff available: 7 Ph.D.'s; 10 M.S.'s; 2 B.S.'s
Quality Assurance/Quality Control Procedures and Capabilities
149
-------
QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? Yes
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? No
Archive facility for the data generated? Yes
Computer-generated data? Yes Are these computer systems vabdated? not reported
150
-------
Virginia Institute of Marine Science
R. Huggett/M. Roberts Testing site: VIMS, Division of Chemistry
VIMS, Division of Chemistry and Toxicology
and Toxicology Gloucester Point, VA 23062
Gloucester Point, VA 23062 Hours: not reported
804-642-7236
Bioassays Conducted
Types of bioassays: VIMS conducts acute and chronic bioassays on fish and chronic bioassays on invertebrate
species. (See tables for description of exact test capabilities.)
Future plans: none beyond those described
Field sampling capabilities: vessels of various sizes; otter trawl nets; Smith Maclntyre grabs
Analytical Chemistry Facilities in Support of Bioassay Procedures
Major area of work or specialty: organic chemicals/varies
Major equipment available:
electron microscopy: 1 TEM
1 SEM
spectrophotometry: 2 MS
chromatography: numerous GC
HPLC
Additional Resources
Computerized information retrieval services available: Hewlett Packard Data System for analytical data reduction
Computers and software available: PC's; Prime 9955
Staff available: 1 Marine Scientist A; 1 Laboratory Specialist A
Quality Assurance/Quality Control Procedures and Capabilities QA/QC program? Yes
In compliance with Good Lab Practices requirements of TSCA and FIFRA of August 17, 1989? Yes
Follow study protocols? Yes Have a complete set of Standard Operating Procedures? Yes
Archive facility for the data generated? Yes
Computer-generated data? partly Are these computer systems validated? Yes
151
-------
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