The United States
Environmental Protection
Agency
Office of Research and
Development (MD-591)
Cincinnati, OH 45268
EPA/600/F-98/007
May 1998
http://www.epa.gov
vvEPA Research within the
Ecological Exposure
Research Division
Cincinnati, Ohio
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Ecological Exposure Research Division within EPA's
Office of Research and Development
Office of Resources
Management and Administration
Office of the Assistant
Administrator
for Research and Development
Office of Science Policy
Office of Research and
Science Integration
Atmospheric
Modeling
Div. (RTF, NC)
Human
Exposure &
Atmospheric
Sciences Div.
(RTF, NC)
Microbiological
& Chemical
Exposure Assessment
Research Div.
(Cincinnati, OH)
Ecosystems
Research Div.
(Athens, GA)
Environmental
Sciences Div.
(Las Vegas, NV)
--''^&m]:ijM^jsE:
- National Water
Cover: A stream bioassessment team conducting electrofishing in support of Regional Environmental Assessment Program
(REMAP). Map (inset) shows a digital elevation model of Mid-Atlantic Integrated Assessment area where EERD-based field
crews sampled approximately 1,000 streams from 1993-1997.
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NATIONAL EXPOSURE RESEARCH LABORATORY
Ecological Exposure Research Division
Cincinnati, Ohio
The Ecological Exposure Research Division (EERD), within the National Exposure
Research Laboratory, develops suites of biological indicators to describe the condition of
watershed ecosystems. The Division also develops indicators to quantify the extent, magnitude
and sources of exposure of ecosystem components to chemical, biological and physical stres-
sors. Charged to facilitate ecological risk assessments by the Office of Research and
Development's Strategic Plan, the Division conducts laboratory and field studies aimed at
providing research products that enable the Agency to conduct predictive and retrospective
exposure assessments. To achieve these broad goals, the Division follows a sequential pro-
cess:
1) Development of measures of ecological condition and exposure linked to known
chemical, physical or biological stressors;
2) Field validation of indicators, testing their practicality, signal-to-noise ratio, and
fidelity;
3) Diagnostic analyses employing combinations of measurements to reveal patterns
characteristic of stressors;
4) Application of tools in both local and national programs to characterize exposure and
evaluate ecosystem risks.
To conduct this research the Division employs ecologists, toxicologists, and biochemists to
conduct studies on a variety of scales of biological organization ranging from the molecular
and organismal level to the population, community, and ecosystem scale. The Division
provides biological reference materials to the Regions and states and assists with performance
evaluation studies to validate and standardize Agency analytical methods.
M. Kate Smith, Ph.D.
Division Director
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Structure & Functions
Who We Are
The Office of Research and Development
(ORD) of the U.S. Environmental Protection
Agency is organized into five National
Research Laboratories or Centers. The
primary mission of the National Exposure
Research Laboratory (NERL), which is
headquartered in Research Triangle Park,
North Carolina, is to conduct research on the
extent and nature of exposure of humans and
the nation's ecosystems to environmental
pollutants and other stressors. Research within
NERL is conducted in six divisions at four
locations across the U.S., as shown in the
organizational chart on the inside front cover.
The research program of the Ecological
Exposure Research Division (EBRD) is
divided among three Branches: the Molecular
Ecology Research Branch, the Ecosystems
Research Branch, and the National Water
Quality Assurance Programs Branch.
EBRD staff comprise a unique combina-
tion of scientists with both expertise in
laboratory science and strength in field
ecology. They are, for example, unique in
exploring the interface between ecology and
molecular biology, and they are the first to
attempt an ecoregional scale assessment of a
molecular indicator in fish. The EBRD staff
currently consists of 43 EPA employees and 7
employees hired under the Senior Environ-
mental Employment Program. Five research
fellows and 28 onsite contract employees also
support the Division's research program. The
academic qualifications of the EPA technical
staff include 19 doctoral, 9 masters, and 11
bachelors degrees in disciplines of chemistry,
biology, ecology, toxicology, botany, zoology,
mathematics, and statistics. The Andrew W.
Breidenbach Environmental Research Center
(AWBERC) facilities provide laboratory
support for exposure research on a wide range
of experimental animals including mammals,
fish, and terrestrial and aquatic invertebrate
species. The AWBERC animal facility is fully
certified by the American Association for the
Accreditation of Laboratory Animal Care.
Molecular Ecology
Research Branch
The Molecular Ecology Research Branch
develops molecular biological and biochemical
indicators of exposure. Indicator research
spans multiple levels of biological organiza-
tion from the subcellular (tissue concentrations
of biochemicals) to the population (genetic
characteristics of a population). Genetic
indicator methods include molecular finger-
printing techniques for analyzing genetic
diversity of populations, gene expression
systems for studying exposure to high priority
chemical contaminants, cytogenetic and
molecular methods for assessing exposure to
genotoxic agents, and techniques for measur-
ing DNA and protein adducts as molecular
dosimeters in exposure monitoring studies.
Research focuses not only on the development
of innovative indicators using laboratory
methods, but also on the evaluation of these
methods in the field to establish reference
levels and to investigate uncertainties with
their application. The developed and field-
validated indicators can then be used to learn
about the condition and vulnerability of the
Nation's ecosystems.
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Structure & Functions
Ecosystems Research
Branch
The Ecosystem Research Branch conducts
research to develop methods to identify
important environmental stressors and to
diagnose their sources. Measurements of
biotic assemblages (e.g., index of biotic
integrity) and functional attributes (e.g.,
nutrient cycling) of streams are used to assess
the condition of stream ecosystems. Commu-
nity-level structural and functional indicators
are also used to estimate the intensities of non-
chemical environmental stressors (e.g., habitat
loss). A major emphasis of the Branch is on
the analysis of combinations of measurements
to form indicator patterns. These patterns,
termed stressor signatures, help to better
understand cause-effect relationships and can
be used to prioritize likely stressors and their
sources. Research on ecosystem restoration is
developing methods to measure the efficacy of
stream restoration based on population and
community responses to known disturbances.
National Water Quality
Assurance Programs Branch
The National Water Quality Assurance
Programs Branch (NWQAPB) currently is
charged with conducting laboratory perfor-
mance evaluation (PE) studies to support the
various water programs administered by the
states and EPA under the Clean Water Act.
The Branch conducts the Alternative Test
Procedure Program, providing a means to
evaluate and recommend new technology for
use in evaluating environmental exposure
studies. Other quality assurance areas studied
include drinking water certification activities,
such as onsite evaluation of EPA Regional
Laboratories, and conducting laboratory
certification training courses for state and
federal laboratory auditors. Non-commercially
available chemical and microbiological quality
control samples and standards are provided to
the environmental analytical community in
order to meet the needs of the Agency's
human and ecological exposure programs.
The NWQAPB is currently undergoing a
major transition involving its responsibilities
and research activities. First, the quality
assurance activities have entered their last
round and are proceeding on schedule so that
they can be brought to conclusion by the end
of fiscal year 1999. Second, the Branch is
transferring the information necessary to
enable the private sector to provide perfor-
mance evaluation studies, and is transferring
the responsibility for reference materials and
other quality assurance samples, including
some of the materials themselves, to other
governmental entities. Third, the Branch is
embarking on a new research direction
involving the development of chemical
indicators for measuring contaminant levels
and exposure of biota inhabiting stream
ecosystems.
Initial projects
include methods
development and
analyses of
streams for
nutrients and trace
metals, as well as
soils and sedi-
ments and various
tissues for
pesticides and
their metabolites.
EEFtD scientist
performing quality
assurance analyses on
standard materials used
in nationwide perfor-
mance evaluation
studies. Gas
chromatography is used
here to separate and
quantify pesticides (red
peaks on monitor).
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Structure & Functions
Red blood cells from
bluegill sunfish following
electrophoresis in an
agarose gel. The
"comet" appearance of
damaged cells, with red
DNA particles trailing
behind, is indicative of
exposure to a genotoxic
pollutant.
Focus of EBRD's Research
Program
Guided by ORD's Strategic Plan, the
Ecological Exposure Research Program of
NERL has developed a common concept
across all Divisions of measuring and model-
ing ecosystem vulnerability at multiple scales.
The primary benefits to be derived from this
research program will be to:
• provide the techniques, and in some cases
the data, necessary to determine the
relative risk of multiple stressors, at
multiple scales, for the Agency and
environmental managers at all levels of
government;
• develop an improved understanding of
ecological exposure, including better
measurement methods and models; and
• develop exposure assessment methods for
use by local, regional and national
environmental managers.
In accordance with the goals of this
research program, the Ecological Exposure
Research Division has developed a research
strategy with two main parts. In one, indicators
are developed to measure the condition of
valued ecological resources and the exposures
that place them at risk. In the second, the
ability to measure and analyze exposures to
environmental stressors is used to estimate the
vulnerability of ecosystems, monitor and
assess the environment, and recommend and
evaluate the options for ecosystem restoration.
The long term goal of this research will be to
develop suites of indicators that allow for the
diagnosis of existing exposures, and to
develop sampling and analysis strategies that
permit this knowledge to be used at regional
and local scales. Data from this research are
used by the Agency to diagnose ecological
problems or to forecast future ecological
conditions.
The research strategy is depicted in
Figure 1 (at right). The first section of the
diagram depicts indicator development, a
process that leads to a characteristic of the
environment that provides evidence of (1) the
condition of the valued resource, (2) the
magnitude of stress, (3) habitat characteristics,
or (4) degree of exposure to the stressor.
Indicator development is a sequential
process. Measurement Development is
primarily a laboratory process and creates the
method for measuring the indicator. Indicator
Evaluation is the research that defines the
detection limits and uncertainties of the
indicator in a real world situation. Field-
evaluated indicators can then be combined
with other indicators or used directly to assess
whether ecosystems are stressed. Ecological
Criteria Development denotes the ranges of
values that occur in the environment and can
be used to set achievable protection and
restoration goals. Diagnostic Analyses
establish patterns of indicators that reveal
sources and critical stressors.
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Research Highlights
In the second part of the diagram, two
assessment scales are shown, "Source-Biased
Studies" (stressor- or effects-initiated studies
generally at a local scale) and "Regional
Studies" (ecological value-initiated assess-
ments at a large scale). Empirical data on
vulnerable ecosystems is used in conjunction
with other data to complete ecosystem
analyses especially at the watershed or
regional scale. This is shown in the diagram
as an arrow leading to "Ecological Risk
Analysis."
Regional priorities and management goals
serve as the driving force for focusing research
on particular indicators or indicator types.
New indicator methods for measuring condi-
tion and exposures are needed when there is
no existing measurement for important
ecological stressors or sources, or when
existing methods have a high level of uncer-
tainty and/or low sensitivity. Sometimes the
methods and tools are already available, but
modifications for specific applications are
required. In some cases, a stressor or source is
already known to be important and needs to be
quantified. In other cases, the stressors or
sources are not known, but the resource may
be degraded or believed to be at risk. For
these situations, ecological receptors (e.g., fish
with tumors) are studied for clues to under-
stand the causes of their condition. Sometimes
a single line of inquiry can demonstrate cause
and thereby reveal the key stressor. More
often, several indicators need to be analyzed
Ecological Research within the
Ecological Risk Assessment Process
Regional Priorities
*
Ecological
Criteria
Development
Figure 1. Indicator
development and
research on vulnerable
ecosystems.
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Research Highlights
FURTHER READING
Lattier, D. L., Gordon,
D.A., Silbiger, R.N.,
McCormick, F., and
Smith, M.K. (1996)
Extremely RAPD
Fingerprinting: A Versatile
Tool for Population
Genetics, in Techniques in
Aquatic Toxicology (Gary
K. Ostrander, Editor), CRC
Press, Inc., Lewis
Publishers, Boca Raton,
Florida, pp 569-575.
DNA fingerprinting
technology is used in the
development of methods
for the regional scale
evaluation of the genetic
diversity of aquatic
species.
collectively so that likely stressors or sources
can be identified, quantified and ranked. When
combinations of measurements are studied,
patterns or signatures can emerge that are
characteristic of stressors. An example of this
diagnostic approach that distinguishes toxics
from nutrient-type stressors is illustrated in a
conceptual model that uses measurements from
fish assemblages (Figure 2, next page).
EBRD develops indicators that can be
used to describe the characteristics and integrity
of aquatic ecosystems and receptors at several
levels of biological organization and across
multiple geographic scales. The focus is on:
• Indicators of population vulnerability,
• Indicators of community condition, and
• Indicators of habitat and ecosystem
condition.
We also develop new indicators or analyze
existing measurements to characterize the
nature of sources of stressors to aquatic
ecosystems. Emphasis will be on:
• Indicators of chemical stress, and
• Indicators of habitat and ecosystem
degradation.
Some of the current research projects in the
Division that employ these indicators are
described below.
Population Vulnerability: Molecular
Indicators of Genetic Diversity in Fish
Advances in the fields of basic and forensic
molecular biology have enabled EBRD re-
searchers to develop indicators for the measure-
ment of both specific chemical stressors and of
general vulnerability to further stress. A
primary interest has been the development of
"genetic similarity indices" which measure
levels of genetic homogeneity within given
populations of any species of plant or animal.
DNA fingerprints are generated and indices
are calculated for populations of organisms
collected from exposed or potentially exposed
areas. The indices can be compared to those
from reference populations to determine their
vulnerability to continued or new exposure.
This research addresses the question of
whether quantitative measures of genetic
diversity are useful as indicators for targeting
potentially sensitive (genetically homoge-
neous) populations.
Current research is aimed at development
and optimization of various methods for
evaluation of genetic similarity and heterozy-
gosity of particular species to be sampled in
regional vulnerability studies. A computerized,
laser-scanning, fluorometric gel reader and an
automated genetic analyzer enhance the
sensitivity and reliability of the DNA finger-
printing methods used for diversity analyses
and allow the high volume of sample process-
ing required for regional scale genetic diver-
sity studies. Optimization includes maximizing
the conditions for generating and sensitively
measuring genetic polymorphisms as well as
ensuring highly reproducible fingerprints.
Methods are also needed for evaluation of
genetic similarity indices on a regional scale.
The creation of relational databases will allow
the correlation of these indices with landscape
characteristics (e.g., percent land in agricul-
ture), chemical measurements, ecotoxicity
tests, and biomarker measurements to yield
regional exposure characterizations for
comparative risk assessments.
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Research Highlights
UJ
.AGRICULTURE.
RESIDENTIAL DEVELOPMENT _
INDUSTRY,
Livestock
Pasturing &
Feedlots
Lawn and
Garden
Maintenance
L
Decreased:
#sp.
# Intolerant spV
Sensitive sp.
# Sunfish sp.
Decreased:
#sp.
# Individuals
Biomass
Increased:
% Tox. Tolerant sp.
% DELT Anomalies
Fish Assemblage
Community Condition: Biotic
Assemblages as Indicators of Stream
Condition
The Ecosystem Research Branch conducts
stream bioassessment research to comprehen-
sively evaluate the biological components of
aquatic ecosystems. The goal of this research
is to promote an integrated, multidisciplinary
approach to the assessment of the structural
and functional integrity of stream ecosystems.
Research areas include the responsiveness of
fish, macroinvertebrate and periphyton
community structure to natural and anthropo-
genic disturbance, and the development of new
methods for detection of ambient water
toxicity and sediment toxicity. This research
has provided methods development, indicator
research and technical support to the USEPA's
Environmental Monitoring and Assessment
Program (EMAP), Regional EMAP projects
and the Superfund Program.
Habitat and Ecosystem Condition:
Functional Indicators as Measures of
Ecosystem Sustainability
How can risk managers know if their
actions are helping or harming the resources
they aim to protect? One method is to measure
Figure 2. Conceptual
model depicting
relationships among
land uses, sources,
stressors and valued
resources. The gray
lines trace toxic
stressors. Blue lines
trace nutrient stressor
pathways. Diagnostic
signatures appear in
boxes in lower half of
diagram.
FURTHER READING
Silbiger, R.N. etal. (1998)
Preliminary Studies on
Population Genetics of the
Central Stoneroller
(Campostoma anomalum)
from the Great Miami
River Basin, Ohio. Envir.
Monitoring Assessment 51:
481-495.
McCormick, F.H., Hill,
B.H.,Parrish,L.P.,and
Willingham,W.T.(1994)
Mining Impacts on Fish
Assemblages in the Eagle
and Arkansas Rivers,
Colorado. Journal of
Freshwater Ecology 9:
175-179.
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Research Highlights
FURTHER READING
Hill,B.H.,Lazorchak,
J.M.,McCormick,F.H.,
and Willmgham, W.T.
(1997) The Effects of
Elevated Metals on Benthic
Community Metabolism in
a Rocky Mountain Stream.
Environmental Pollution
95:183-190.
Scientists from the
Ecosystem Research
Branch conduct
bioassessment research
in conjunction with the
Arkansas and Eagle
River Superfund
Projects.
the loading of stressors including metals,
sediments, and nutrients. Another is to
measure reductions in the toxicity of the media
being cleaned, such as soil, sediment or water.
Although valuable, these measures fall short of
providing the information really needed to
know if the ecosystem is functioning as it
should. Risk managers need quantifiable
measures of ecosystem function and
sustainability before they can chart the
progress, and ultimately the accrued benefit, of
stream restoration efforts.
Ecosystems are complex, self-regulating,
functional units. Unlike communities and
populations which are structurally defined,
ecosystems are defined by rates and processes.
Functional indicators are those metrics which
directly or indirectly measure energy flow and
material cycling within ecosystems.
The goal of this research is to develop and
evaluate functional indicators of stream
ecosystems. A secondary goal will be to assess
spatial and temporal variability of these
functional indicators in streams among and
between ecoregions. The central hypothesis is
that functional indicators are sensitive to
watershed stressors, such as nutrient enrich-
ment, siltation, and mine drainage, and
changes in function will be indicative of
changes in ecosystem integrity associated with
these stressors.
Riparian Zone Indicators
Using the Little Miami River watershed as
an example (see p. 14), NERL scientists are
conducting research on riparian zones. These
vegetated stream banks serve as the interface
between the water body and the surrounding
terrestrial ecosystem. They serve an important
buffering function by reducing non-point
source pollution, excess light, and sediment.
They also serve as highly productive areas
both for human activities (e.g., agriculture)
and for wildlife conservation. The extent of
riparian zone protection and the features that
contribute most to the protective attributes of
riparian zones are not well characterized.
Indicators to assess the protective potential of
different riparian zones are also untested. This
research will develop criteria to judge riparian
zone condition and relate it to the condition of
the stream it borders. This project aims to
address this need and to develop such criteria
by: 1) developing methods for assessing
riparian resources over a watershed scale; 2)
determining which features of riparian zones
(e.g., width, completeness, and vegetation) are
most important in their various functions; and
3) confirming the validity of these features by
developing a model which relates stream
biological integrity and water quality with
riparian zone features. By providing linkages
between riparian resources and biological
integrity/water quality, the role of the riparian
zone can be documented, and its rational use
in watershed restoration can be suggested.
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Research Highlights
Indicators of Chemical Stress:
Biological/Chemical Indicators of
Contaminants in Streams
Reference values for exposure of fish to
contaminants are needed to be able to quickly
determine whether a site is highly contami-
nated. EERD researchers are developing
guidance to help set such reference values. In
one study, measurement of exposure to oil and
combustion product contamination was
selected to develop these methods for aquatic
organisms. Reference values for naphthalene
(NAPH), associated with oil contamination,
and benzo[a]pyrene (BAP), associated with
combustion by-products, were determined for
fish in Ohio streams by measuring bile
metabolites. Reference concentrations for
these two types of biliary polycyclic hydrocar-
bon (PAH) metabolites were determined for
white suckers. Bile from white suckers was
collected from first- to third-order streams in
the Regional Environmental Monitoring
Assessment (REMAP) study of the Eastern
Corn Belt Plain (ECBP) ecoregion at statisti-
cally selected sites (triangles on map) and
from third or higher order streams in conjunc-
tion with the Ohio EPA (OEPA)
Biomonitoring Program which sampled sites of
concern and selected reference sites (circles on
map). There were significant statistical
differences between fish from the statistically
selected REMAP sites and those from OEPA
reference sites which were selected on profes-
sional judgement. The REMAP study showed
that 7.5% of first- through third-order streams
in the ECBP ecoregion exceeded both the
NAPH and BAP criteria values. More than half
the OEPA non-reference sites exceeded criteria
values for both types of metabolites. The goal
• OEPA Non-reference Sites
0 OEPA Reference Sites
A REMAP Sites
| | Eastern Corn Belt Plains
(ECBP) in Ohio
Ohio
Rivers in ECBP
FURTHER READING
Lin, E., Cormier, S. M.,
and Torsella, J. (1996) Fish
Biliary Polycyclic
Aromatic Hydrocarbon
Metabolites Estimated by
Fixed Wavelength
Fluorescence: Comparison
with HPLC-Fluorescent
Detection. Ecotoxicol.
Environ. Saf. 35: 16-23.
White sucker were
collected by
electro-fishing during the
summers of 1992-95
from streams in Ohio.
Bile samples were
analyzed fluorimetrically
for benzopyrene and
naphthalene metabolites.
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Research Highlights
Molecular techniques
such as reverse
transcription-PCR are
used to measure the
expression of genes that
are associated with
exposure of aquatic
organisms to endocrine
disrupting compounds.
Purple-blue DNA bands
appear under
fluorescent light.
of finding meaningful exposure criteria was
successfully accomplished in this study. The
guidance provided by this study can be used to
develop criteria of exposure for other contami-
nants to other wildlife and other ecosystems.
Indicators of Habitat and
Ecosystem Degradation: Stressor
Signatures of Habitat Degradation
Among Metrics from Fish, Benthic
Macroinvertebrate, and Periphyton
Assemblages
The use of aquatic organisms as indicators
of human disturbance dates back as far as the
mid 1800s. Today, biological monitoring and
assessment programs are in place in 41 states.
Assessment results are used in decision
making on attainment of aquatic life uses
required by the Clean Water Act. They also
aid in non-regulatory decisions related to water
resource management and community-based
ecosystem protection.
Distinctive patterns in fish and inverte-
brate assemblages can differentiate among
different anthropogenic sources. Such evi-
dence can be used to indicate which sources
cause a change in the stream community.
While field observational data cannot be used
alone to prove causality, data with diagnostic
power can provide important support for
causal inferences. A conceptual model has
been developed and is being used to guide
analysis by indicating different response
patterns that may be expected and the particu-
lar measurements which may have discrimina-
tory value.
Analyses will be conducted for the mid-
west using a database constructed in coopera-
tion with the State of Ohio and the Ohio EPA.
This database focuses on sampling sites in the
Eastern Corn Belt Plain ecoregion. In the mid-
Atlantic, data from EMAP sampling in 1993-
1997 and state data from Pennsylvania and
Maryland will be used. While not all mea-
sures are the same or complete for all data sets,
available data types include: fish, benthic
macroinvertebrate and periphyton assemblage
data. Also included are chemistry data on
nutrients and toxics, habitat data, land use
data, point sources and sample locations and
location linkings. Analyses of these data will
yield indicators which may be used to
recognize the relative impact of varied
stressors of human origin. The research will
provide guidance on development of diagnos-
tics for multiple stressors and a practical
example in which those diagnostic methods
are used.
The ability to measure something does not
necessarily make it useful for identifying
vulnerable ecosystems or the exposures that
cause them to be threatened. The process of
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Research Highlights
field evaluation helps to weed out those mea-
surements subject to too much error when used
outside the laboratory. In order to validate the
indicator being developed, questions about the
usefulness and suitability of the indicator need
to be addressed. Among the important
questions are:
• Does the indicator offer a new capability
or offer improved sensitivity, specificity
or robustness over existing methods?
• What are the potential monitoring
applications of the indicator? Can it be
used across various scales of time and
space? Will the indicator be useful for
assessing the vulnerability of ecosystems?
• Have potential customers been identified?
(Who is likely to use the indicator and
who will benefit from its use?)
• Have the limits of detection and variabil-
ity of response of the indicator to known
exposures been adequately defined? Is the
signal-to-noise (S:N) ratio sufficiently
high to be useful in field applications?
Does the S:N vary from site to site? What
factors besides exposure affect the S:N
(e.g., health, age of organisms, seasonal
influences)?
• Has the indicator been observed to be
linked to exposure to a given stressor by a
conceptual model?
• What are the logistical constraints
associated with deployment of the
indicator for field evaluations (i.e., time,
cost, tissue requirements)?
• Have appropriate controls been incorpo-
rated into the study design (e.g., appropri-
ate reference sites)?
• Have data quality assessment issues in
study design been adequately addressed
(e.g., sampling strategies, number of
organisms needed)?
Once an indicator has been validated, it is
ready for field application. Several ongoing
projects in the Division illustrate how indica-
tors are being deployed.
Program Offices
EBRD is interacting with the Office of
Water and the Office of Pesticide Programs
within OPPTS.
• The Office of Water develops "biologic
criteria" (e.g., Biological Criteria -
Technical Guidance for Streams and
Small Rivers - May 1996 - EPA-822-B-
96-001) and has adopted five objectives
for meeting the Clean Waters and Safe
Drinking Water national goals (http://
www .epa.gov/OWOW/indic/
tblconthtml). To measure the progress
toward these objectives, the OW
has encouraged the development
of a series of indicators. EBRD is
developing indicators that will
measure: (1) the exposure to
chemical stressors and habitat
alteration, (2) species at risk due
to reduced genetic diversity, and
(3) biological integrity.
• Ecological risk assessment efforts
within the Office of Pesticide
Programs also benefit from
EBRD research. OPP interests
such as "aggregate exposure
assessment" and the development
of indicators for monitoring
pesticides in ground and surface
waters are being addressed. Fate
Blood cells from fish are
being applied to gel-
coated slides in
preparation for
measurement of DNA
damage.
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Research Highlights
Table 1.
Representative
products from the
Indicator Research
Program of benefit
to EERD clients.
and transport of modern, moderately
persistent pesticides are being studied as
part of the development of indicators of
riparian zone quality.
Further interactions with both Offices will
allow applied scientific contributions from
EBRD's watershed and regional indicator
evaluation studies. Some of the other EERD
products and clients are listed in Table 1.
Regional Vulnerability Assessment
(ReVA)
The ReVA program is a coordinated
research effort involving all the divisions of
NERL. The goal of the program is to develop
the next generation of measurements, models
and tools necessary to assess the vulnerability
of ecoregions to the stresses imposed by the
simultaneous impacts of such things as wide-
spread urbanization and population growth,
industrial pollution, erosion and chemical
contamination from agriculture, and changes
in regional climate. As the most important
stresses within a region are identified, the
emphasis will be on learning how to project
the consequences of the stresses for the future.
Finally, the researchers will be learning how to
communicate the answers to reducing the
stresses - along with a sense of their reliabil-
ity, their costs, and the risks in failing to act -
to the broad audience of citizens and officials
who bear ultimate responsibility for America's
future.
ReVA is being initiated in conjunction
with the Mid-Atlantic Integrated Assessment
(MAIA) project. The study area of approxi-
mately 162,000 mi2 includes all of PA, MD,
WV, VA, and parts of NC, DE, NY, and NJ.
This study area was selected because of the
availability of extensive spatial data archives
and past and ongoing EMAP sampling efforts.
A key product planned for this effort will be
the development of an atlas describing some of
the important stressors in the Mid-Atlantic
Products
State of Streams report for Mid-Atlantic
Genetic diversity methods and profiles
Reproductive indicators and profiles
IBI, SBI, PIBI, and geographic coverages
Indicators of restored ecosystems
Riparian function index
Vulnerability of Streams report for Mid-Atlantic
Big Darby Creek Watershed Ecological Risk Assessment
Comparison of EMAP and state protocols
Large River Methods Manual
Stressor signatures
Mid-Atlantic Streams Fish Tissue Contaminant Report
Client
ORD (ReVA)
ORD (ReVA)
ORD (ReVA)
ORD (ReVA); ORD (EMAP)
ORD (Ecorestoration)
Office of Water; Office of Pesticides
ORD (ReVA)
ORD (Risk Assessment); Office of Water
ORD (EMAP); Region and States
Region and States; Office of Water
Office of Water
ORD (ReVA)
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Research Highlights
Region. These include the impacts of mining,
agricultural chemicals (both pesticides and
fertilizers), ground-level ozone, wide-spread
conversion of land to suburban development,
increases in damaging ultra-violet sunlight
from loss of stratospheric ozone, and soil
erosion and its impact on rivers and streams.
The atlas will show where in the Region
stresses are accumulating as a result of human
demands on the environment. This will help
federal and state officials begin to focus their
efforts on those most-endangered localities
that can benefit most from targeted regulatory
controls and ecological restoration programs.
Ecologists in the Ecological Exposure Re-
search Division are the principal investigators
for the mine drainage stressor and stream
ecosystem receptors.
EMAPand REMAP
The Environmental Monitoring and
Assessment Program (EMAP) is designed to
use environmental monitoring data from
multiple spatial and temporal scales to assess
ecological condition and forecast potential risk
to our natural resources. Since 1992, scientists
in the Ecosystem Research Branch have served
as EMAP-Surface Waters leads for the fish,
macroin vertebrate, and periphyton community
indicators, sediment toxicity and fish tissue
contaminants indicators and benthic commu-
nity metabolism indicator. Under the auspices
of the EMAP, REMAP (see below) and TIME
(Temporally Integrated Monitoring Effort)
projects, approximately 1,000 sites were
sampled in the Mid-Atlantic Region.
The Regional Environmental Monitoring
and Assessment Program (REMAP) represents
a new partnership among EMAP, EPA's
Regional offices, other federal agencies, and
states. REMAP was initiated to test the
applicability of EMAP's statistical design and
indicator concepts to assess ecological
condition at smaller geographic scales and in
shorter time frames than the national EMAP
program. Scientists in the Ecosystem Re-
search Branch have served as Project Officers
or technical liaisons for several REMAP
projects including:
Region I: Fish Tissue Contamination in the
State of Maine;
Region II: Characterizing Sediment Quality
in the New York/New Jersey Harbor
System;
Region III: Surface Water Quality Indicators
in the Central Appalachian Ridges and
Valleys Ecoregion;
Region V: Developing an EMAP Signature
for a Rare and Imperiled Ecosystem,
Assessing Corn Belt Rivers and Streams;
Region VI: The Status of Habitat Degradation
and Biotic Integrity in East Texas Streams;
Region VII: Measuring the Health of Fisher-
ies;
Region VIII: Assessing Water Resources in
the Mineralized Area of the Southern Rocky
Mountains Ecoregion;
Region IX: Assessing Aquatic Ecosystems in
a Highly Modified, Agriculturally Influ-
enced Environment: California's Central
Valley.
Risk Assessment: Big Darby Creek.
Ecological Risk Assessment Case
Study
The Big Darby Creek watershed is a
freshwater, aquatic ecosystem, encompassing
557 square miles in central Ohio. The
watershed is an example of a high-quality
FURTHER READING
Environmental Monitoring
and Assessment Program
Surface Waters: Field
Operations and Methods
for Measuring the
Ecological Condition of
Wadeable Streams. (1997)
Edited by J.M. Lazorchak
and D.J. Klemm. EPA/620/
R-94/004,
Yeardley, R.B., Lazorchak,
J.M.,andPaulsen,S.G.
(1998) Fish Tissue
Contamination in
Northeastern U.S. Lakes:
Evaluation of an Approach
to Regional Assessment.
Environ. Toxicol. Chem.
(In Press).
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Research Highlights
Geographic information
systems (GIS) are used
by EERD researchers to
analyze the sources and
distribution of stressors
at watershed and
regional scales. Maps
generated through GIS
provide easy color-
differentiation of areas of
high exposure to
stressors.
ecosystem which is relatively free of pollu-
tion. This exceptional stream is threatened by
an assortment of stressors originating with the
day-to-day activities of people in the water-
shed. The risk assessment will provide
objective results for understanding how land-
use decisions can affect the integrity of the Big
Darby Creek's complex ecosystem. The risk
assessment relates land use and the associated
stressors to effects on native species and
ecological integrity. The assessment is
designed chiefly for the people who live in the
watershed since their actions will have the
greatest effect on the future of the watershed.
The Federal and Ohio Environmental Protec-
tion Agencies also benefit by learning better
ways to scientifically predict environmental
risks.
Ecorestoration: Predicting stream
ecosystem restoration potential
EERD is developing the scientific
framework for establishing stream ecosystem
restoration potential based on observed stream
ecosystem responses to known disturbance
gradients. The relationship between stream
ecosystem structure and function will be
established by quarterly monitoring of six
stream reaches in the Monday Creek water-
shed of southeastern Ohio and monitoring
every four months of 13 stream reaches in the
Arkansas River in the Southern Rockies of
Colorado. Both stream systems are impacted
by metal contaminated sediments associated
with coal or hardrock mining. Monday Creek
is impacted additionally by acidic mine
drainage.
This research will provide a comparison of
structural and functional responses of stream
ecosystems to metal sediment contamination
associated with mining. During the project,
the effectiveness of mine drainage
remediation for restoring stream ecosystem
structure and function will be assessed. The
overall goal is to develop a means for rapidly
collect in-stream ecological information for
diagnosing existing ecosystem structure and
function and relating it to restoration poten-
tial. This information will help scientists and
managers prioritize their restoration needs
within a watershed.
Near Laboratory Ecological
Research Assessment (NLERA):
The Little Miami River Watershed
Ecological scientists from each of the
four NERL facilities nationwide have selected
a nearby (100 km) watershed (i.e., NLERA) to
serve as a focal point for the respective
division's ecological field research. The
general idea behind the NLERA concept is to
provide, for each facility, a field research area
that is well characterized (e.g., in terms of
geology, physiography, hydrology, and
ecology) and, at the same time, is near enough
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Research Highlights
to facilitate cost-effective logistics for field-
based efforts in ecosystem research. In the
course of conducting their own field research,
the Divisions will seek to participate in
ongoing efforts to find solutions to the
recognized problems in these watersheds by
seeking out other watershed researchers from
all governmental, academic and corporate
organizations. The goal will be to form
collaborative arrangements which will help
leverage resources and provide more compre-
hensive approaches to dealing with watershed
problems on a basin-wide scale.
The EBRD has selected the Little Miami
River watershed as its NLERA site. The Little
Miami River (LMR) is a 105.5-mile long
tributary of the Ohio River located in south-
western Ohio. The Little Miami River
watershed covers 1,757 square miles and
traverses two ecoregions (Eastern Corn Belt
Plains and Interior Plateau). Sections of the
LMR mainstem flow through steep-sloped,
forested gorges containing some of the most
scenic riverine habitat in Ohio. The Ohio
Environmental Protection Agency (OEPA) has
determined that the longest section of Excep-
tional Warmwater Habitat (EWH) in the state
is located on the LMR main stem. Accord-
ingly, the LMR has been designated as a State
and National Scenic River.
The predominant land use in the upper
LMR basin is agricultural. However, in the
lower sections the watershed is experiencing
increasing residential and commercial devel-
opment of land in the riparian corridors.
EBRD's ecological research in the LMRW
will assess the impacts of agriculture, increas-
ing development and population pressures in
Features of the
landscape along the
Little Miami River, such
as the width of the green
vegetated stream bank
(riparian zone), are
being evaluated for their
effectiveness in reducing
levels ofstressors in the
watershed. The
watershed is also
serving as a field
research area for EERD
scientists to test and
evaluate indicator
methods.
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Research Highlights
EERD scientists use
confocal microscopy and
flow cytometry to
develop subcellular
exposure indicators
diagnostic of specific
chemical stressors.
Different colors on the
monitor show varying
intensities of indicator
fluorescence.
the watershed. A key focus will be the
assessment of ecological integrity using
ecological indicators (based on community
and ecosystem measures) and biological
markers (based on biochemical and physi-
ological endpoints) as tools for verifying
stressor loads and for the diagnosis of caus-
ative elements in stressor profiles. Associa-
tions between biotic integrity and abiotic
stressors are dependent on development of
methods for assessment of riparian zone
quality and the application of these assessment
tools over a larger scale via the application of
Geographic Information Systems (GIS). GIS
will be used to overlay stressor loads, ecologi-
cal receptors, and riparian zone quality with
chemical, physical and biological indicators of
stream quality.
SERDP: Ecological Biomarkers:
Monitoring Wild Fauna at Department
of Defense (DoD) Installations
The Strategic Environmental Research
and Development Program (SERDP) has
supported development of technology to assist
DoD in establishing a quantitative data base
for the assessment of ecological risks associ-
ated with nitroaromatic munitions chemicals.
EERD scientists have shown that
nitroaromatic munitions compounds and their
by-products bind to the DNA and protein of
exposed animals in a manner proportional to
the dose received (see Fig. 3). These biologi-
cal markers of exposure can be used to
quantify the actual dose of contaminant that
the animal is receiving from its environment
under field conditions. This research is
innovative in that, typically, biologically
derived information about ecological impacts
has been limited to the utilization of exposure
models in conjunction with measurement of
environmental contamination levels (e.g., soil
or water concentrations). Biological markers,
in contrast, provide direct, predictive and
diagnostic information on the actual exposure.
Thus, biological markers can be used to focus
remedial efforts where they are most beneficial
for the protection of wildlife; e.g., on contami-
nants that are biologically available, which
bioaccumulate, and which are biologically
active (e.g., toxic). This new technology is
being evaluated via a field study at the SERDP
demonstration site at the Volunteer Army
Ammunition Plant, Chattanooga, TN, in
collaboration with scientists from the U.S.
Army Center for Health Promotion and
Preventive Medicine, Aberdeen Proving
Grounds, MD. This approach to exposure
assessment should permit DoD to determine
actual exposures under field conditions and
thus to prove or disprove the cause-effect
relationships associated with nitroaromatic
munitions and also guide cleanup and restora-
tion activities.
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Research Highlights
Other Activities of the Division
Laboratory Certification Program
Purveyors of potable waters are required
under the Safe Drinking Water Act (SDWA)
to monitor their water for regulated contami-
nants at prescribed frequencies. The samples
collected by this monitoring must be analyzed
in a "certified" laboratory which will assure
the production of known quality data. These
data will then be used by the state to judge the
quality of water produced by these drinking
water treatment plants. The approval of
laboratories is the basis of the certification
effort carried out by the USEPA and states.
The design and offering of training
courses to support laboratory certification is
the primary responsibility of the National
Water Quality Assurance Programs Branch.
The Branch offers courses in chemistry and
microbiology laboratory certification annually.
These courses are designed to instruct USEPA
Regional and state persons in the correct
manner of certifying laboratories analyzing
samples to comply with the SDWA. In answer
to special requests, these training courses are
offered at remote sites other than Cincinnati.
The NWQAPB also is responsible for the
onsite evaluation of EPA regional laboratories.
Teams of EPA personnel from Cincinnati
perform onsite evaluations of the regional
laboratories in both the areas of chemistry and
microbiology. All aspects of laboratory
operations are audited, including sample
preparation, chain of custody procedures,
holding times, analyses and reporting. A
formal report is provided to the Regional
Laboratory Director itemizing the results of
the audit and the contaminants for which the
laboratory is certified.
Pesticide Repository
Standards of pesticides are available
generally on a continuous basis to selected
laboratories determined by either the US
EPA's Office of Pesticides Programs or the
Cover
— »»™™- Carnivorous
x . -"^Herbivorous Wildlife
*$; Wildlife
Exposure
Assessment
Soil
Blood
Sample
and
Analysis
Munitions Exposure Level
Figure 3. The
movement of
nitroaromatic munitions
by-products (e.g., TNT)
through the ecosystem
can be monitored by
measuring the chemical
reaction products
(adducts) they form with
blood proteins.
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Research Highlights
US Food and Drag Administration. Normally
20-50 mg amounts are provided with a percent
purity value if available. Material Safety Data
Sheets are also provided. Each eligible
laboratory has a catalog which lists the
compounds by generic name as well as major
trade name or common name. A request form
is also provided with the catalog. Requests are
shipped according to Department of Transpor-
tation regulations by an EPA contractor,
normally within two weeks. Overnight express
service is available for emergency shipments.
Biological Methods and Manual
Development
Environmental measurements are required
to determine the quality of ambient waters and
the character of stressors. NERL-Cincinnati
conducts research to develop and evaluate
analytical methods for the identification,
enumeration and evaluation of aquatic
organisms exposed to environmental stressors
and to correlate the exposure with effects on
chemical and biological indicators. NERL-
Cincinnati scientists serve as the Agency's
scientific lead for the following stream and
source monitoring indicators: fish,
macroinvertebrates, periphyton, zooplankton,
functional ecosystem indicators, water and
sediment toxicity and fish tissue contaminants.
As part of their indicator lead responsibilities
NERL-Cincinnati scientists prepare and update
field and laboratory protocol and methods
manuals for these indicators. They also
provide technical assistance to the EPA
Regions, Program Offices and states on the
implementation and interpretation of these
manuals. The current manuals and protocols
prepared by NERL-Cincinnati scientists are:
Environmental Monitoring and Assessment
Program Surface Waters: Field Operations
and Methods for Measuring the Ecological
Condition ofWadeable Streams. EPA/620/R-
94/004. 1997.
Short-term Methods for Estimating the
Chronic Toxicity of Effluents and Receiving
Waters to West Coast Marine and Estuarine
Organisms. EPA/600/R-95/136. August, 1995.
Short-term Methods for Estimating the
Chronic Toxicity of Effluents and Receiving
Water to Freshwater Organisms. EPA/600/4-
91/002. July, 1994.
Short-term Methods for Estimating the
Chronic Toxicity of Effluents and Receiving
Water to Marine and Estuarine Organisms.
EPA/600/4-91/003. July, 1994.
Methods for Measuring the Acute Toxicity of
Effluents and Receiving Waters to Freshwater
and Marine Organisms. EPA/600/4-90/027F.
August, 1993.
Fish Field and Laboratory Methods for
Evaluating the Biological Integrity of Surface
Water. EPA/600/R-92/111. March, 1993.
Macroinvertebrate Field and Laboratory
Methods for Evaluating the Biological
Integrity of Surf ace Waters. EPA/600/4-90/
030. November, 1990.
For additional information about
U.S. EPA's exposure research,
visit the following internet web
site: http://www.epa.gov/nerl/
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Scientific Expertise
Telephone*
Area of Expertise
Office of the Division Director
M. Kate Smith, Director 7577
Robert Graves, Associate Director 7197
F. Bernard Daniel, Senior Science Advisor 7401
Molecular Ecology
Research Branch 7419
Greg P. Toth, Chief
Jeno P. Bercz
Lina W. Chang
Suzanne A. Christ
Susan Franson
Denise A. Gordon
David Lattier
John R. Meier
Tirumuru V. Reddy
Annette Roth
Richard Silbiger
Paul A. Wernsing
Thomas K. Wessendarp
Ecosystem Research
Branch 7907
Susan M. Cormier, Chief
Gary B. Collins
Florence A. Fulk
Brian H. Hill
Donald J. Klemm
Development of aquatic indicators
Aquatic chemistry methods; aquatic program manager
Wildlife toxicology; riparian zone ecology
Biochemistry; reproductive biology
Clinical biochemistry and pathology; toxicology;
analytical cytochemistry and cytology
Cellular and genetic toxicology; aquatic indicators of
genotoxicity
Reproductive biology; developmental biology
Ecology; environmental statistics
Molecular biology; genetic diversity
Molecular biology; gene regulation; environmentally
induced gene expression
Genetic and environmental toxicology
Biochemistry; DNA and protein adducts
Biochemistry
Molecular biology; gene regulation; population
genetics; genetic diversity
Genotoxicity assessment and methods development
DNA isolation/purification
Cell biology; ecology; exposure criteria; risk
assessment; source and stressor diagnostics
Management of scientific data; biological indicators
and methods
Environmental statistics
Ecosystem ecology; stream and wetland ecology;
aquatic botany
Aquatic macroinvertebrates; aquatic toxicology; field
and laboratory methodology
*(513) 569-xxxx
(continued on next page)
-------
Scientific Expertise
Telephone*
Ecosystem Research
Branch (continued)
James M. Lazorchak
Edith L. C. Lin
Frank H. McCormick
Benjamin H. McFarland
Mary Schubauer-Berigan
Bhagya Subramanian
National Water Quality
Assurance Programs Branch
Raymond J. Wesselman, Acting Chief
Daniel F. Bender
Paul W. Britton
Richard L. Carr
Louis E. Feige
Tamara Goyke
Patricia A. Hurr
Donald L. Klosterman
Raymond L. Loebker
Charles W. Moench
Natalie Murff
James W. O'Dell
7325
Area of Expertise
Ecotoxicology; aquatic ecology; field and laboratory
biological methods; aquatic chemistry
Biochemical indicators; exposure criteria
Stream ecology; ichthyology
Effect of chemical stressors on aquatic plants and
diatoms
Environmental risk assessment; statistics
Exposure criteria; risk analysis and database
management
Analytical chemistry
Methods development; quality assurance; disinfectant
analysis
Statistics, quality assurance procedures, laboratory
performance evaluation criteria
Organic analytical chemistry
GC/MS and pesticide chemistry
Microbiology performance evaluation studies
Laboratory certification
Organic analytical chemistry
Trace metal analysis using ICP-argon emission
spectroscopy
Inorganic analysis by ion chromatography
Coordination of performance evaluation studies
Alternate test procedures; inorganic methodology
*(513)569-xxxx
-------
U.S. EPA Cincinnati
Kentucky HT
o
Greater
Cincinnati
Airport
275^
DIRECTIONS TO EPA
From Airport:
1-275 East to 1-71/75 North.
Stay on 75 N when it splits from 1-71 just past
the bridge.
Exit #3 - Hopple St. (on left).
Left at light at end of exit ramp.
Follow Martin L. King Dr. to EPA.
Printed on Recycled Paper
-------
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