MUL 77- YEAR PLAN FOR

SAFE PESTICIDES/SAFE PRODUCTS
            2007-2015
  Office of Research and Development

 U.S. Environmental Protection Agency
          December 2006

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                                Administrative Note

The  Office of Research and Development's (ORD) Multi-Year Plans (MYPs) describe what
research ORD proposes to conduct over the next 5-10 years in specific high priority areas. The
MYPs serve  four principal purposes, to:  1) describe the future directions of the research
programs, 2) present the  anticipated significant  outputs  of the research, identifying which
laboratory/center  will be responsible and the timeframe  in which  it will be completed,  3)
communicate the  research plans within ORD and with stakeholders and clients, and 4) identify
the significant accomplishments  and outcomes of previously conducted research.   Multi-year
planning permits ORD to consider the strategic directions of the Agency and how research can
evolve to best contribute to providing the scientific underpinnings for the Agency's mission of
protecting human  health and the environment.

MYPs are intended to be "living documents." ORD updates MYPs on a periodic basis to reflect
the current state of the science,  resource availability,  and  Agency priorities.   This MYP was
reviewed by ORD's Science Council in October, 2006, and approved in November, 2006. The
research directions and past accomplishments described in the MYP will serve as the foundation
for an external peer review by the Safe Pesticides/Safe Products Subcommittee of ORD's Board
of Scientific Counselors (BOSC) to be held February 7-9, 2007.

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               Safe Pesticides/Safe Products Research Program
                     Contributors to the Multi-Year Plan
Research Planning Team:
ORD Members:
Alva Daniels (NRMRL)
Jack Fowle (NHEERL)
Elaine Francis (IOAA)
Ross Highsmith (NERL)
Susan Laessig (NCER)
Sue Makris (NCEA)
Jacqueline McQueen (OSP)
Sonya Robinson (ORMA)
Bill Russo (NHEERL)
Greg Sayles (NRMRL)

Program and Regional Office Members:
Cathy Fehrenbacher (OPPT/OPPTS)
George Hess (Region 7)
Randy Perfetti (OSCP/OPPTS)
Kathleen Rafaelle (OPP/OPPTS)
Phil Sayre (OPPT/OPPTS)
Jennifer Seed (OPPT/OPPTS)
Ingrid Sunzenauer (OPP/OPPTS)
Royan Teter (Region 7)

Key ORD Investigators:
Appendix V

Science Council Reviewers:
Jeff Morris, Associate Director for Science (OSP)
Bob Olexsey, Acting Associate Director for Ecology (NRMRL)
Chris Saint, Acting Associate Director for Health (NCER)
Michael Slimak, Associate Director for Ecology (NCEA)

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                                TABLE OF CONTENTS




I. Introduction                                                                  5




II. Background                                                                  7




III. Relationship of EPA's Research to that of Other Organizations                     10




IV. Progress to Date/Changes from Previous Version                                 13




V. Long Term Goals                                                            14




VI. Description of the Flow Diagrams and SP2 Research Program                     18




VII. Relationship to Other Multi-Year Plans                                        42




VII. Relationship to the Research and Development (R&D) Investment Criteria          44




IX. Communications                                                            44




X. References                                                                  45




Appendix I. Potential Additional Research if Resources Increased 10-20 Percent        46




Appendix II. Flow Diagrams Figures 3-5                                           48




Appendix III. Annual Performance Goals/Annual Performance Measures Tables 1-3      52




Appendix IV. Details on Research Themes                                         62




Appendix V. Key ORD Investigators                                               93




Appendix VI. Accomplishments                                                  95




Appendix VII. Acronyms                                                       100

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I. INTRODUCTION

The US Environmental Protection Agency's (EPA) Office of Research and Development's (ORD)
Multi-Year Plan (MYP) for Safe Pesticides/Safe Products (SP2) describes the research program that
is specifically designed to address the problem-driven science needs of the Office of Prevention,
Pesticides and Toxic Substances (OPPTS).  The purpose of the SP2 Research Program is to provide
OPPTS with the scientific information it needs to reduce or prevent unreasonable risks to humans,
wildlife, and non-target plants  from exposures to pesticides, toxic chemicals,  and  products  of
biotechnology.  The SP2 Research Program specifically addresses OPPTS' high priority research
needs that are not addressed by any  of ORD's other research programs. The research program is
focused on:   1)  providing  OPPTS  with  predictive tools  for prioritization of regulatory  data
requirements; enhance the interpretation of data submitted as part of the regulatory process in order
to  improve  human  health and ecological risk assessments; provide  targeted,  multidisciplinary
research in response to OPPTS requests on filling critical data gaps for specific individual or classes
of pesticides  and  toxic  substances  that are of high  priority;  2)   developing the scientific
underpinnings necessary  to transform ecological risk assessments to a more realistic, spatially-
explicit probabilistic basis where effects on wildlife and non-target plants can be evaluated as to
their impacts at the wildlife  population and plant communities levels;  and 3) providing  the tools
necessary for OPPTS  to update its requirements for  submissions of registrations for products of
biotechnology and the scientific foundation to help OPPTS interpret data submitted.

The scope of the SP2 research program has been developed in partnership with OPPTS. OPPTS has
the responsibility of carrying out the mandates of numerous laws including the  Toxic Substances
Control Act (TSCA), Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA), and Food
Quality Protection Act (FQPA)  As such, OPPTS plays a leading role in regulatory risk assessment
in EPA and, consequently, a number  of its research needs are similar to those of other EPA Offices.
Therefore, to a certain extent, research  conducted  in direct support of other EPA Offices (e.g.,
Office of Water) as well as research conducted under ORD's core human health, human health risk
assessment,  and ecological protection research programs  are  providing many  of the scientific
methods and models needed by OPPTS.

In order to avoid duplication of research, ORD asked the senior OPPTS leadership to articulate the
major scientific uncertainties in implementing their regulatory programs. ORD did a cross walk of
OPPTS' strategic needs with the research directions of all of its research programs articulated in
MYPs.  Figure 1 illustrates the alignment of OPPTS needs with  ORD research.  This led to the
identification of major areas that were not addressed by other research programs and where SP2
resources for research should be focused. In instances where research is being conducted through
several research programs on a particular need, the research is coordinated and complementary, and
not duplicative. Descriptions of the research that complements the SP2 program can be found in the
MYPs    for    the    following     research    programs:         Endocrine    Disrupters
(www.epa.gov/osp/myp/edc.pdf), Drinking Water (www.epa.gov/osp/myp/dw.pdf), Human Health
(www.epa.gov/osp/myp/eHH%20MYP%20Final.pdf),    Human   Health    Risk   Assessment
(www.epa.gov/osp/myp/HHRA.pdf),  and Ecological  Research (www.epa.gov/osp/myp/eco.pdf).
(See Section VII for more details on relationship of SP2 research with that in other ORD programs).

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          OPPTS PROGRAM
               NEEDS
OPPTS SCIENCE
    NEEDS
ORD MULTI-YEAR
     PLAN
 ORD
 RESEARCH
AREAS
              FQPA - OSCP •

        FQPA, TSCA, FIFRA  •
         OSCP, OPPT&OPP
                 FQPA-OPP-

             HPV/PMN-OPPT_
        Pesticides/lnerts-OPP'
              EDCs-OSCP
                 SAP-OPP-

        OPP, OPPTS & OSCP •

               OPPT&OPP-

                FQPA-OPP-

                FIFRA-OPP-
EDC - Screening
& Testing

Testing Protocols/Data
Interpretation

Agg/Cumul Risk:
Suscept Subpops

Prioritization & Ranking

Ecological Risk

Biotech

Perfluorinated
Chemicals

Pesticides in DW

Spray Drift




EDCs

EDCs & Safe
Pesticides/Products

EDCs
Human Health

Computational Tox,
SP2 & EDCs

Eco & Safe
Pesticides/Products

Eco & Safe
Pesticides/Products

Safe
Pesticides/Products

EDCs
Drinking Water

Safe Pesticides/Products
(previous MYP)
                                            • S & T Methods Dev.

                                            _ New Testing
                                             Paradigm
                                             Dev. Impacts;
                                            •Children's Health;
                                             Agg/Cumul Risk
                                            • Computational Tox
                                             Wildlife/Plant Commun.
                                            • Effects Assm'ts;
                                             Exposure Assm'ts
                                            _ Allergenic Response/
                                             Eco Fate/Risk
                                             Toxicity, PBPK,
                                            • Degradation, Fate &
                                             Transport
                                            _ Characterization &
                                             Treatment
                                            | Exposure &
                                            ' Treatment
Figure 1. Alignment of OPPTS Needs with ORD Research

The  research under the  SP2 program is consistent with the recommendation of the National
Research Council (NRC,  1997) report Building a Foundation for Sound Environmental Decisions,
that the Agency should maintain a balanced program of "core and problem-driven research." The
NRC indicated that problem-driven research is targeted at understanding and  solving particular,
identified environmental problems.  The SP2 program consists mainly of problem-driven research
and provides the balance to,  and is coordinated  with, the "core" research in the complementary
programs.
ORD's SP2 research falls under EPA's Strategic Plan (2006-2011)  Goal 4 Objective 4. Goal  4,
Healthy  Communities and Ecosystems, commits the Agency to protect, sustain,  or restore the
health of people, communities, and ecosystems using integrated and comprehensive approaches and
partnerships.  Objective 4 commits the Agency to Enhance Science and Research, by pledging the
following: through 2011, identify and synthesize the best available scientific information, models,
methods, and analyses to support Agency guidance and policy decisions related to the health  of
people, communities, and ecosystems.  EPA's Strategic Plan further commits the Agency to focus
research  on pesticides and chemical toxicology;  global change; and  comprehensive, cross-cutting
studies of human, community and ecosystem health.	

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As  noted in the Agency's  Strategic Plan  (www.epa.gov/ocfo/plan/2006/entire_report.pdf), a key
component of  protecting the health  of  people, communities,  and ecosystems  is  identifying,
characterizing, and reducing any unreasonable risks presented by the thousands of chemicals on
which  the US population depends. For example, chemical  and biological pesticides help meet
national and global demands for food; provide effective pest control for homes, schools, gardens;
and control animal vectors of disease.  Every day the general public in the US comes into contact
with industrial and  commercial chemicals that are in  products throughout our  homes  and
workplaces.  The  SP2 research program  is providing OPPTS with the tools  it needs  to make
decisions about these chemicals.

The SP2 program is  one of a few ORD research programs that include diverse multi-disciplinary
efforts in the areas of human health, wildlife, and plants and cuts across  the risk assessment/risk
management paradigm.   The complexity  of the  research in  this program  is reflected in the key
science questions  it is  addressing.   In  several  specific  areas  (e.g.,  perfluorinated chemicals,
biotechnology), researchers are working in partnerships across disciplines to address the complex
critical science needs.   Such  concerted multi-disciplinary efforts will  enable  us  to  achieve the
Agency's Goal 4.4 Objective, as it relates to SP2, to "conduct research  that  contributes to the
overall health of people,  communities, and ecosystems."

The SP2 MYP arrays ORD's research program for the period 2007-2015 and revises  and updates
the  previous MYP prepared in 2003 (US EPA, 2003).  The SP2 MYP provides a focused research
framework  and direction that reflects available ORD scientific capabilities and capacity.  The
research  described  in  this  MYP  assumes  annual  intramural  and  extramural  resources  of
approximately 126 FTEs and $24.8 million, including payroll, travel and operating expenses.

Decisions regarding  the conduct of specific research efforts  under SP2 are based on the ORD's
strategic and annual  planning processes, which involve input and prioritization  of research by the
ORD  and Program and Regional Office  members of research planning  teams as well as other
Agency (e.g., OPPTS, ORD)  senior managers,  risk assessors, and stakeholders.   The methods,
models, and data developed through the SP2 research program are externally peer-reviewed and
widely disseminated. The SP2 research program will undergo periodic external review by the Safe
Pesticides/Safe Products Subcommittee of ORD's Board of Scientific Counselors (see Section VIII
for additional details).

II. BACKGROUND

OPPTS' priorities and  regulatory programs

The authorities and responsibilities of OPPTS are mandated primarily by TSCA (15 USC (C. 53)
2601-2692.  1976 http://www.access.gpo.gov/uscode/titlel5/chapter53_.html), FIFRA  (7 U.S.C. s/s
136    et   seq.,    1996,    http://www.access.gpo.gov/uscode/title7/chapter6_.html),   FQPA
(http://www.epa.gov/pesticides/regulating/laws/fqpa/), and the Pollution Prevention Act (42 USC
13101,  1990,  http://www.epa.gov/opptintr/p2home/p2policy/actl990.htm).  These  statutes direct
OPPTS to perform a wide  variety of activities with the goal of protecting human health and the
environment.  OPPTS is authorized by TSCA, FIFRA, and FQPA, for example, to request data in
order  to:  1) evaluate the  potential effects of  industrial chemicals, pesticides, and  products of
biotechnology on human health and the environment, 2) discern when additional data are needed, 3)
                                             7

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set allowable levels of exposure or releases to the environmental that are protective of human health
an the environment, and 4) determine whether risk management approaches are needed, and if so,
which ones.

It follows, from their regulatory mandates, that there are several generic  key areas of scientific
needs that OPPTS has had historically:   1) the development of scientifically  sound methods and
models according to which OPPTS would in turn require the chemical and pesticide industries to
submit data  in compliance with TSCA  and FIFRA, 2) the necessary data and tools  needed by
OPPTS to interpret the data submitted by industry and from other sources to evaluate the risks of
chemicals, pesticides,  and products of biotechnology, 3) frameworks under which assessments for
human and ecological risks would be conducted, and 4) targeted data gaps on a single chemical or
class of chemicals needed to complete an upcoming risk assessment and/or inform risk management
decisions.

The first three areas are being met by multiple research programs through ongoing longer-term
research efforts that are focused on OPPTS' greatest needs that  evolve over time, as priorities shift
and science  advances.  Some of the research to address these areas, especially  if it has broad
applicability to other Program and Regional Offices, is conducted under other research programs
mentioned previously (also see Section VII).  On the  other hand, research that is of specific value to
mainly OPPTS, such as developing methods and models that it would implement under TSCA and
FIFRA, targeted research to support their specific risk assessment/risk management decisionmaking
are conducted under the SP2 research program.

The fourth area, by the nature of the more immediate need to meet a deadline for a particular
assessment, is  met through shorter-term research efforts, and is largely conducted only through the
SP2 research program. This requires the Research Program to be sufficiently flexible to adapt and
accommodate  the  periodic urgent specific requests while maintaining the ongoing longer-term
efforts.
For the last three decades, ORD research, under the
SP2 Research Program and its predecessors,  has
provided OPPTS with continually improved testing
methods  that  OPPTS,  in turn,  uses to  produce
guidelines for  the industrial chemical  and pesticide
industries to  follow in  generating  toxicity  and
exposure data.   Therefore, for the relatively small
investment that  ORD has made  in developing  test
methods,  the  Agency has received  hundreds  of
millions of dollars' worth of data.  Also historically,
ORD research has been providing OPPTS with the
needed tools to  assist them in interpreting  the data
once they are  submitted and incorporating  the data
into their risk  assessments.  The  research described
in this MYP continues to  provide the  same type of
commitment and support.
Long Term Goals.  Briefly, research is
used in:
1 - prioritization of testing requirements,
enhanced interpretation of data to improve
human   health   and   ecological   risk
assessments, and decisionmaking regarding
specific individual or classes of pesticides
and  toxic  substances that  are  of  high
priority.
2 - probabilistic risk assessments to protect
natural  populations  of  birds, fish,  other
wildlife, and non-target plants.
3 - decisionmaking related to products of
biotechnology.

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The three identified overarching long-term science needs have been  structured  as the long-term
research goals for ORD's SP2 Research Program. The first goal is to build on the decades of test
method development for assessing the risks of chemicals, to develop genomic and computational
methods  for prioritization of  regulatory data requirements,  to facilitate the  interpretation of
submitted data in risk assessments, and  conduct short-term research to address targeted needs for
upcoming specific  risk assessment/management decisions.  The second  goal is to develop the
scientific underpinnings  necessary to transform  ecological risk  assessments  to  a more realistic,
probabilistic basis where effects can be  judged by their impacts  at the population level and  plant
community level. The third goal is to provide the underlying science needed to evaluate products of
biotechnology.  See Section V for a more detailed description of the SP2 Long Term Goals (LTGs).

Figure 2 provides a conceptual framework for the SP2 research program, which links the resources
to the outputs and programmatic outcomes to reduce or prevent risks to humans, wildlife, and non-
target plants from pesticides, toxic  substances,  and products of biotechnology.  The principal client
for the SP2 research program is OPPTS.  However, the research from this program is also of value
to scientists in  other  ORD  research  programs,  ORD's National  Center  for Environmental
Assessment, and risk assessors in  other Agency Program and Regional Offices, the states,  other
federal  agencies,  international organizations, the  regulated  community,  and  the academic
community. These stakeholders use the products of this research, in line with the LTGs, as the
scientific foundation for:  1) Prioritization of testing needs, enhanced  interpretation of data, and
decisions on targeted chemicals or classes of chemicals; 2)  Probabilistic risk assessments for better
informed decisionmaking in protecting  natural populations of birds, fish  and other wildlife, and
plant  communities; and  3) Decisionmaking related to  products  of biotechnology.   Progress is
measured by the extent to which methods, models and/or data from the SP2 research program are
actually used in peer-reviewed risk  assessments and other decisionmaking. The use of SP2 research
products by OPPTS and others will contribute to decisionmaking related to reduction or prevention
of exposures or releases of potentially harmful pesticides, toxic substances,  and products of
biotechnology into the environment.

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                  Safe Pesticides/Safe Products
                      Research Program  Logic





Inputs &
Re-
sources

•Strategic
guidance
•Research
Coordination
Teams: client
needs, research
questions & ,
priorities,
outcomes, multi-
year plan
•Partnerships
•ORD
Intramural &
Extramural
(STAR)
Research
Resources
•Program
level funding
&FTE









»








Effective Outreach and Transfer to
Intended Clients
Key research outputs are peer-reviewed & transferred to
clients as evidence for rulemaking and through publicat ons,
briefings, technical consultatons, scientific workshops,
conferences, public n^etings, interagenc^tworking groups
T 1

Research
Activities
Conduct effects,
exposure, & risk
management
•esearch for:
•Prioritization of
esting
equirements,
enhanced
nterp relation of
data&
decisonmaking for
ligh priority
argeted
chemicals/classes
•Probabilistic risk
assessments to
)rotect natural
)opulations of birds,
ish and other
m Id life
•Dec sionmaking
elated to products
if b otechnology

4











»








Research
Outputs
Research
orovides efficient
& effective
methods,
models & data
:or:
•Prioritizing testing
•equirements
•Interpreting data
For use in human
and ecological risk
assessments
•Decisionmaking
•egarding specific
ligh priority
chemicals/classes
•Spatially explicit
srobabilistic
ecological risk
assessments
•Assessing
sroducts of
si otechnology









>








Intended
Clients
EPA
(OPPTS,
Regions,
OW)
Regulated
agrichemical
and industrial
chemical
industries
Scientific
Academic
Community
Federal
Agencies
International
Organizations
States
Tribes













>







sn




art-Term
Program Mission
To provide timely, leading-edge research
products supporting sound scientific
decisions by the Office of Prevention,
Pesticides, and Toxic Substances


Outcomes
OPPTS and/or
other


organizations
use ORD's
research as the
scientfic
foundation for:
•Prioritization of
testinc
needs,

enhanced
interpretation of
data & decisions
on targeted
chemical/class-
specific
•Probabilistic risk
assessments to
protect natural
populat ons of
birds, fish , other
wildlife and plant
communities
•Decisonmaking
related to products
of biotechnology

444

4










»








Inter-
mediate
Outcomes

Pesticides, toxic
substances,
and products of
biotechnology
that pose an
unreasonable
risk to humans
and/or wildlife
are identified
and their
release to the
environment is
prevented or
reduced














»








Environ-
mental
Outcomes

Reduction or
prevention of
exposures or
releases of
potentially
harmful
pesticides,
toxic
substances,
and products
of
biotechnology
into the
environment














»








Long -Term
Outcomes


Risk to humans,
wildlife, and
plant
communities
from pesticides,
toxic
substances, anc
products of
biotechnology is
reduced or
prevented






444
External Influences that may Impact the SP2 Research Program
Congressiona appropriations and Administration budget decisions, changes in EPA science priorities, changes in EPA regulatory requirements,
availability of investment capital, consent agreements
Figure 2.  Logic Model for SP2 Research Program

III. RELATIONSHIP OF EPA'S RESEARCH TO THAT OF OTHER ORGANIZATIONS

Research outside of EPA

While several US Federal Agencies either conduct or support research in the area of test methods'
development, their scope or approach differs from EPA's (see Long Term Goal 1).  For example,
the National  Toxicology Program (NTP)  coordinates  toxicology  testing within  the  federal
government, conducts an interagency validation process for alternatives to whole animal tests, and
convenes panels to  develop risk assessments on targeted chemicals/classes of chemicals.  The
National Institute of Environmental Health Sciences (NIEHS)  conducts and supports laboratory-
based research to reduce the burden of human illness from environmental causes and has a Small
Business Innovation Research (SBIR) program that supports the development of new toxicological
testing approaches and promotes the technology transfer of new knowledge about mechanism for
toxicity    into   applied    testing    methodologies   (www.niehs.nih.gov/external/resinits/ri-
lL_htm;www.niehs.nig.gov/oc/factsheets/analt.htm).    There  is  an  interest both within NTP,
specifically its Interagency Coordinating Committee  on  the Validation of Alternative Methods
(ICCVAM) and NIEHS, as there is at EPA, for  the development of test methods that use fewer
animals. ICCVAM evaluates and validates new test methods for implementation.  NIEHS is using
their intramural and extramural programs (e.g., National Center for Toxicogenomics) to develop
                                         10

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alternative methods and models including computerized modeling and predictive systems, tissue
cultures, transgenic cells and animals, invertebrate species, and non-mammalian vertebrate species
such as fish. Scientists from ORD's Comp Tox and SP2 Research Programs are collaborating with
NIEHS on some of these efforts.

Various Agencies and scientific organizations conduct research to develop probability-based
exposure models for ecological risk assessments complementing ORD's research (see Long Term
Goal 2). For example, researchers at the Department of Energy's Argonne National Laboratory and
Oak Ridge Laboratory have developed a number of probabilistic fate and transport, food-web, and
aquatic models for evaluating potential risks to wildlife and aquatic receptors, for a range of
environmental contaminants, at or near contaminated sites. The US Army sponsored the
development of two wildlife exposure models (Spatially Explicit Exposure Model and FISHRAND-
Migration) for improving the realism of terrestrial wildlife exposure models and predicting organic
chemical uptake based on prey consumption and food web dynamics, respectively. The National
Oceanographic and Atmospheric Agency is conducting research into tools for expressing ecological
risk probabilistically. The United Kingdom's Department for Environment, Food, and Rural Affairs
sponsors research to develop probabilistic approaches to reduce uncertainty in pesticide fate
modeling.  German researchers are collaborating on a research project (XPROB) to evaluate the use
of human exposure factors in exposure modeling and provide guidance on probabilistic modeling.
Arysta LifeSciences Corporation sponsored the development of the Probabilistic Exposure and Risk
model for FUMigants (PERFUM) to address bystander exposures to fumigants following
agricultural applications. CANTOX Environmental and Bayer Crop Science developed the
probabilistic exposure model Granular Pesticide Avian Risk Assessment Model (GranPARAM) to
estimate bird exposure to granular pesticides.

ORD participates in several interagency  activities relevant to SP2  research.   ORD co-chairs the
Committee on the Environment and Natural Resources and its Toxics and Risk Subcommittee under
the  auspices of the Office  of  Science and Technology Policy (OSTP).  ORD is a member of the
Biotechnology Steering Committee under OSTP's Committee  on Science.  ORD is also a member
of the Biotechnology Research Working Group (BRWG) and the Agricultural Biotechnology Risk
Analysis (AGRA) Task Group, both working groups under the Steering Committee.  Through these
panels, research  activities  are shared across the agencies to facilitate collaborations and avoid
redundancy.  AGRA has developed a framework  for federal research on agricultural biotechnology
risk assessment and an inventory of ongoing  federal research (the public document is in the process
of being cleared for release).   As a result  of overlaying these two efforts, high priority research
gaps have been identified.  ORD's  SP2 research program addresses several of these areas (see Long
Term Goal 3).  In addition, ORD participates on  the United States-European Union Biotechnology
Task Force. All of these efforts help ensure that ORD's SP2 research program is not duplicative of
those conducted elsewhere.

The  Organization for  Economic Cooperation  and Development (OECD)  consists  of 30 member
countries and plays a  prominent role in fostering "good governance in the public service and in
corporate  activity"  (www.oecd.org).    Under the  OECD,   EPA  (mainly   OPPTS  and  ORD)
participates on  many of their working  groups  aimed at  promoting  the  development  and
harmonization of chemical testing  guidelines and risk assessment approaches.  For example, ORD
has taken the  lead within the OECD in  revising the testing  guidelines  on  developmental
neurotxocity (TG 426).  While the OECD itself does not  conduct research it promotes  scientific
                                           11

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innovation and encourages cross-member collaboration on research leading to the development of
new testing guidelines and paradigms; and improved risk assessment and risk management tools.
For example, ORD is participating in collaborative efforts through OPPTS with the OECD and the
International Programme on Chemical Safety (IPCS) of the World Health Organization (WHO) on
protocol  development  and the application  of toxicogenomics  in chemical  assessments.   The
OECD's and IPCS'  main areas of specific interest in the application of toxicogenomics include: 1)
development of effective, increasingly efficient, and rapid approaches for the hazard screening of
large number  of chemicals,  2)  improving understanding  of cross-species sensitivity to  facilitate
cross-species  extrapolation,  3)  development  of biomarkers, 4)  harmonization  of  acceptance
requirements among across member countries.  These efforts reflect ORD's current interests and
activities in the SP2 research program as well.  ORD's participation in these efforts demonstrates
the international recognition accorded to its scientists.

The  chemical  and pesticide industries develop the chemical-specific  data  that are submitted to
OPPTS in accordance with regulatory  requirements.  In addition, the industry  conducts other
research.  The American Chemistry Council (ACC) is a trade association of more than 190 member
companies that represents the majority of the manufacturers of industrial chemicals in the US. ACC
coordinates the chemical industry's research and testing programs. In 1999,  the ACC initiated the
Long-Range Research Initiative (LRI) to sponsor research aligned with health and environmental
issues.   Their latest five year  strategy (2005) identifies three focus areas:  improved methods,
susceptible populations, chemicals in the environment   Commonalities exist  among these topics
with the general  objectives of the research in SP2, the  Human Health,  Human Health Risk
Assessment, Endocrine Disrupters, and  Ecological  Protection Research Programs.  In 2005-2006
the LRI is supporting 55 projects (www.uslri.org).  They have ongoing research activities in many
areas that complement EPA's intra- and extramural programs, e.g., studies  on testing  and testing
methodology,  susceptible populations,  mechanisms of action, epidemiology, animal toxicology,
wildlife studies, aquatic toxicology, environmental exposure, and environmental chemistry.

CropLife America (CLA) is the national trade association representing the plant science  industry.
One   aspect   of   their   activities   is   the   Crop   Protection   Research   Institute
(www.croplifefoundation.org).   They provide  funding for the economic  analysis of agricultural
pests, pest management, and pesticide use and regulation.  In particular they support the  National
Pesticides Use database.

Research conducted in EPA

No other programs have similar goals, in terms of scope and mission, as the  SP2 research program
that  provides OPPTS with the tools it  needs to carry out  its regulatory  mandates.  EPA's SP2
research is multi-disciplinary, including:  1) research across all aspects of the risk  assessment/risk
management paradigm, i.e., in effects, exposure, risk assessment,  and risk  management; and, 2) as
related to humans, wildlife, and plants. No other single organization has such an extensive portfolio
of ongoing research in  providing methods,  models,  and  data for reducing  scientific uncertainty
regarding pesticides, toxic substances,  and products of biotechnology. Comparison of  potential
benefits is conducted  from a  scientific perspective  through coordinating with  other  research
programs, participation at national and international  scientific fora, and keeping abreast of state of
the science.  EPA's  SP2 program includes many areas that are of unique importance in helping the
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OPPTS meet its legislative mandates, such as requiring industry to submit data on pesticides, toxic
substances, and products of biotechnology. The SP2 program also includes other research areas that
serve to improve the basic scientific understanding regarding these agents that OPPTS and other
parts  of the Agency need to evaluate data  submissions, conduct risk assessments,  and to make
informed management decisions.  Furthermore, ORD's intramural program is complemented by an
extramural program implemented through the Science to Achieve Results (STAR) program.  ORD
participation on interagency  and international fora provides an opportunity  for scientists to stay
aware of research ongoing at other agencies/countries, help to ensure that work is not duplicated,
and help to find potential collaborators.

Focus of EPA's contribution

Priorities for the SP2 research program were assigned based upon an assessment of the importance
of the research to OPPTS, and to a much lesser extent any other Agency Offices, on the magnitude
of the uncertainties in the knowledge base, the sequence of research needed to obtain the final
answer, the possibility  that the research  would result in  a significant  product  for  hazard
identification, risk  characterization  or risk management, the scientific and technical  feasibility of
conducting a successful project, and, finally  either legislatively-mandated or Agency-set time
frames.

ORD has significant expertise in the areas  of toxicology, model development, engineering, and
environmental  exposures,  relating  to both  humans  and  ecological systems, and  in providing
solutions to environmental problems.   ORD  scientists are  respected members of  the  scientific
community and leaders  in their fields of concentration.  Therefore, ORD can make/is making a
significant contribution in the areas of development of more efficient and effective methodologies,
models, and frameworks to  evaluate chemicals, pesticides, and products of biotechnology and
development of approaches to manage any unreasonable risks they pose.  In  addition, through the
STAR program, ORD  is engaging academic institutions to conduct research  in areas that
complement our intramural capability and capacity.

IV. Progress to Date/Changes from Previous Version

Progress to Date
Major accomplishments  of the  SP2   research  program  are  described  in  Appendix VI.
Accomplishments have been  aligned by Long Term Goals (LTGs) which are defined  in Section V.
Where available, a website, where more detailed information can be found, has been provided.

Changes from Previous Version

The SP2  MYP has undergone some changes from the previous (2003) version.  Most of the major
changes that have been made to the SP2 MYP deal with the how the research program is organized
for  presentation.  In addition, the content of the research program has evolved, as it has built upon
its  previous accomplishments and,  through closer collaborations  with  OPPTS, has  refined the
specific research needed. Other changes reflect shifts of resources from extramural  to intramural
research support for the  principle investigators, as well as significant decreases to the overall SP2
budget that have taken place  since 2003.  A list of significant changes in the current version of the
SP2 MYP is as follows:	
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•  The LTG structure has been modified and redefined to make it outcome-oriented; that is, it now
   describes how and by whom the results of the research will be used.
•  LTG 4 has been eliminated due to budget decreases.  Some of the remaining research critical to
   OPPTS has been moved to other LTGs.
•  LTG 1 has been expanded to include the flexibility to conduct multi-disciplinary shorter term
   research  on high priority chemicals or classes of chemicals for OPPTS. This new area includes
   the remaining  research from LTG  4  on  the toxicity  of perfluorinated  chemicals.   The
   perfluorinated chemicals research portfolio has been expanded to take on a multi-disciplinary
   approach and now includes exposure and risk management research because of an increased
   need to understand exposure pathways and fate of this emerging class of chemicals.  Priority
   research  conducted previously under LTG 3, e.g., pesticides in drinking water, has been shifted
   to LTG  1.   LTG 1 has  been expanded  to include computational toxicology  research on
   prioritization, testing, and enhancement of risk assessments that is being  done through the
   National  Center for Computational Toxicology  and the complementary extramural  STAR
   program.
•  LTG 2 has been expanded to include high priority research on the effects of herbicides on plant
   communities, especially as those effects affect the  suitability of the communities as  wildlife
   habitat.
•  The visibility of ORD's research  on biotechnology guided the decision that it should comprise
   its own LTG (3).   Furthermore, as the original research projects from the 2002 initiative come
   to a completion,  the research shifts from  laboratory/center-specific projects toward a  cross-
   laboratory/center integrated project.  Severe budget  reductions to this research, however, have
   led to a much more limited program than described in the previous MYP.
•  Across the board, there are changes to Annual Performance Goals (APGs)  and milestones or
   Annual Performance Measures (APMs).  Some have been added, some have been deleted, and
   some have been delayed.  Deletions include those that have  been met in previous years as well
   as those that will not be done because of budget decreases. Delays in timing are also the result
   of impacts to resources.
•  This version  includes an  Appendix that  highlights the  Accomplishments  of the SP2 research
   program.
•  This version includes an Appendix that provides greater detail to the research theme areas.
•  This version provides improved examples of cross-linkages to other MYPs.

V. LONG TERM GOALS

Long Term  Goals

Long Term  Goal 1:  OPPTS and/or other  organizations  use the results of ORD's research on
methods,  models,  and  data as  the scientific foundation for:   A)  prioritization  of testing
requirements, B) enhanced interpretation of data to improve human health and  ecological
risk assessments, and C) decisionmaking regarding specific individual or classes of pesticides
and toxic substances that are of high priority.
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 The ultimate outcomes are the development of improved methods, models, and data for OPPTS' use
 in  requiring  testing,  evaluating  data,  completing risk  assessments,  and  determining  risk
 management approaches.   More specifically the outcomes are  the  development by ORD and
 implementation by OPPTS of more efficient and effective testing paradigms that will be better
 informed by predictive tools (chemical identification,  improved targeting cost less, less time, and
fewer animals); improved methods by which data from the more efficient and effective testing
paradigms can be integrated into  risk assessments;  and that OPPTS uses the result of ORD's
 multidisciplinary research approaches, that it specifically requests, for near term  decisionmaking
 on high priority individual or classes of pesticides and toxic substances.

 Long Term Goal 2: OPPTS and/or  other organizations use the results of ORD's research  as
 the scientific foundation  for probabilistic risk assessments to protect natural populations  of
 birds, fish, other wildlife, and non-target plants.

 Results of this research will help the Agency meet the long term goal of developing scientifically
 valid approaches to extrapolate across species, biological endpoints and exposure scenarios of
 concern, and to assess spatially-explicit, population-level risks to wildlife populations and non-
 target plants and plant communities from pesticides, toxic chemicals and multiple stressors while
 advancing the development of probabilistic risk assessment.

 Long  Term  Goal  3:   OPPTS  and/or other organizations  use the results of  ORD's
 biotechnology  research as the scientific foundation for decisionmaking related to products  of
 biotechnology.

 OPPTS will use the results from this research program to update its requirements of registrants of
products of biotechnology and to help evaluate data submitted to them.

 Science Questions

 Addressing the following key science questions through the SP2 research program will give OPPTS
 the tools it needs to meet its mandates. Under the LTGs or subparts of the LTGs, the key science
 questions are as follows:

 LTG 1 -Subpart A - Providing OPPTS with  predictive tools for prioritization of regulatory
 data requirements:

    >  What methods are needed for priority setting and screening?
    >  How can existing in silico  and in vitro techniques be harnessed to develop effective and
       efficient screening and prioritization tools?
    >  How  can  existing quantitative   structure  activity  relationship   (QSAR) databases be
       improved?
    >  What endpoints are amenable for the development of in vitro screens?
    >  How can improved understanding of pathways of toxicity lead to improved predictive tools?

 LTG 1 - Subpart B - Enhancing the interpretation of data submitted as part of the regulatory
 process in order to improve human health and ecological risk assessments:

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    >  What methods  are  needed that could enhance the interpretation  of data  from current
       guidelines?
    >  How can current guidelines be revised to  enhance sensitivity and improve data quality and
       interpretation?
    >  Can hypothesis-driven approaches for testing chemicals for multiple toxicity pathways be
       developed?
    >  How can current databases be enhanced and applied to improve access to data for hypothesis
       formulation and test evaluation?

LTG 1 - Subpart C - Providing targeted, multidisciplinary research in response to OPPTS
requests on filling critical data gaps for specific individual or classes of pesticides and toxic
substances that are of high priority:

    >  What methods and  tools  are  needed  for characterizing  the following for perfluorinated
       chemicals?
          o  Effects - toxicity and pharmacokinetics
          o  Exposures - pathways of exposure, environmental degradation, fate and transport
          o  Risk management options
    >  What protocols are needed  for information  on the impact of drinking water treatment
       processes on pesticides?
    >  To what extent, if any, do deck coatings  and sealants reduce dislodgeable residues on the
       surfaces of CCA-treated wood?
    >  How can exposure methods be improved for use in large scale human  studies?
    >  What factors affect the releasability of asbestos?
    >  What chiral  pesticides are good candidates for production of safer, single  enantiomer
       products?
    >  Can fast, simple, inexpensive lead paint test kits be developed quickly?

LTG 2  - Developing the  scientific  underpinnings  necessary to transform  ecological risk
assessments to a more realistic, spatially-explicit probabilistic basis where effects on wildlife
and plants can be evaluated as to their  impacts  at  the  wildlife  population  and  plant
community levels:

    >  What methods are needed for extrapolating toxicological data across wildlife species, media,
       and individual-level  response endpoints?
    >  What methods are  needed for characterizing population-level risks  of toxic chemicals to
       aquatic life and wildlife?
    >  What approaches  are needed  for  evaluating  the  relative  risks from  chemical  and
       nonchemical  stressors on spatially  structured wildlife populations  across large areas or
       regions?
    >  How can methods to assess direct and indirect risks to non-target plant species and plant
       communities from the use of chemical herbicides be improved?
    >  What probabilistic  tools can be used  to  characterize  or  predict the fate and transport of
       pesticides and other environmental contaminants?
    >  How  do environmental  contaminants move through environmental   compartments  and
       become  available for human, aquatic, and wildlife exposures?
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LTG 3 - Providing the tools necessary for OPPTS to update its requirements for submissions
of registrations for products of biotechnology and  the scientific foundation to help OPPTS
interpret data submitted:

    > What are the potential risks of allergenicity to biotechnology products and how can they be
      evaluated?
    > What are the risks to natural ecosystems of gene transfer from engineered organisms to
      natural species in the wild?
    > What methods are needed to mitigate the development of resistance and of gene transfer?

The degree of emphasis for each LTG is based on the following criteria:  1) an assessment of the
importance  of  the  research  to  OPPTS  and other Agency  Offices,  2) the magnitude of the
uncertainties in  the knowledge base, 3) the sequence of research needed to obtain the final answer,
4) the degree of impact the research product(s) are likely to have, 5) their degree of criticality for an
OPPTS  hazard  assessment, risk characterization, or risk management decisions, 6) the intramural
and/or extramural capability  and capacity to ascertain the scientific and technical feasibility of
conducting a successful project, and 7) legislatively  mandated  or other regulatory time frames. The
following Table summarizes the relative emphasis of each LTG over the period of FY2007 through
2015.
LTG
1
2
3
Emphasis from 2007 through 2015
Level and then increasing, when resources are
freed from LTG 2
Level until probabilistic tools are developed
and implemented, then decreasing
Level and then increasing, when resources are
freed from LTG 2
The rationale behind the projected level of emphasis over time is as follows.  Within the next eight
years, it is expected that ORD will have developed the needed tools under LTG 2 for OPPTS to use
in their probabilistic ecological risk assessments. Once those tools are  completed, delivered, and
implemented, the level of effort in that LTG need no longer remain at its current level.  At that time
resources from LTG 2 will be shifted to LTGs 1 and 3 to address projected increasing needs in
those areas.  For example, under LTG 1 there  will  be an increased need for more sophisticated
predictive tools for prioritizing and screening chemicals. Under LTG 3, additional resources will be
needed to restore resources back to their original FY03 level to address the intended goals of the
research program related to agricultural biotechnology needs and to expand to address the growing
need to provide OPPT with tools needed to review products of biotechnology associated with TSCA
mandates.

Successfully addressing these LTGs will require a highly coordinated effort.   The efforts  will be
overseen by the research planning teams under the direction of the National Program Director for
Pesticides and Toxics.  The SP2 MYP, the NHEERL Implementation Plan, the Wildlife Research
Strategy   (www.epa.gov/nheerl/publications/files/wildlife_research_strategy _2_2_05.pdf),   the
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Biotechnology                              Research                               Strategy
(www.epa.gov/nheerl/publications/files/biotechnology _research_program_4_8_05.pdf)     ,    the
Computational                            Toxicology                            Framework
(www.epa.gov/comptox/publications/comptoxframework06_02_04.pdf) and any other strategies or
plans that may be developed around the current topics should be considered collectively when
trying to understand the overall SP2 research program, in order to determine what specific research
the Laboratories and  Centers  will be  carrying out,  and when.   The research  planning teams
recognize  the  dynamic nature  of the SP2 research and,  therefore, will assess the priorities and
sequencing in the  MYP and other related documents periodically, so that  the overall  research
program can be modified as the knowledge base increases, new technologies become available, and
resources shift.

VI. DESCRIPTION OF THE FLOW DIAGRAMS AND SP2 RESEARCH PROGRAM

The flow diagrams (Figures 1-3 in Appendix II) depict the Annual Performance Goals (APGs) that
will support the LTGs, the time frame for their completion, and their interrelatedness. The research
that ORD  is committed to conducting under each of the LTGs is reflected in the APGs and their
respective Annual Performance Measures (APMs) and is  described in  greater detail below and in
attached Tables 1-3 in Appendix III.

The research planning team took the following into  consideration when determining the APGs: 1)
the key questions based on OPPTS' programmatic needs, 2) the research themes that were being
identified  within  NHEERL's  SP2 Implementation Plan  the  Wildlife  Research  Strategy,
Biotechnology Research Strategy, and the Computational Toxicology Implementation Plan, and 3)
an assessment of other ongoing and anticipated ORD research efforts.  As a result, eleven discrete
research areas (APGs) were identified in which ORD, between the intramural and extramural STAR
grants programs, can make a significant  impact in advancing the state of the science on SP2 within
the next 8 years.  Schedules for these research areas were estimated based upon knowledge of: 1)
existing resources, 2) intramural capacity and capability, 3) projected timelines for awarded grants,
4) the complexity of the area, and 5) in some cases, regulatory-driven deadlines.

Each APG has a number of APMs that represent discrete  segments of research to be completed
within the  defined schedule.  The APMs (Tables  1-3 in  Appendix III) will help to determine
progress made towards completing the  APG.    The APMs in the  Tables represent the expected
product from a given research area.  Therefore,  the Tables, for the most part, do  not necessarily
show continual  progress  of a research area  from  start  to  finish,  but rather just  the major
"milestones."   Most of the APMs  are  attributed  to one of ORD's Laboratories  or  Centers.
However,  in an effort to  improve  collaboration  across  laboratories and  centers on  a given
environmental issue, some APMs may represent the products of multiple organizational efforts. It
should be  noted that, for the most part, those that are attributed to NCER are products of STAR
grants.  Some APMs appear in multiple  MYPs (or are leveraged with resources from other MYPs).
It  is  important to note that the Tables  capture  the  APGs and APMs  currently anticipated.   The
research planning team recognizes the need to update the matrix periodically, as new milestones are
anticipated and as emphases shift.
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Please note that the APMs described in this MYP should not be confused with or ever designated as
ORD or Agency APMs used  in  cross-Agency planning and accountability  activities.   It is
anticipated, that APMs from the  MYP  will be aggregated and integrated  to derive  "Annual
Performance Measures" for Agency accountability activities, when required.

The  eleven APGs for the SP2  MYP  are described below.  For each APG, the objective,  the
significance/impact,  and the schedule of the research are described.  It is assumed that the various
APMs, when aggregated, will lead to the achievement of the APG.

A summary of the  science issues  and the research program objectives makes up most  of  the
remaining body of this MYP.  A compendium of what research  would be done if there were more
resources, the timeline  for delivery of  research products by lab oratory/center, more detailed
descriptions of the  research  themes (an aggregation  of multiple research  projects addressing
common key  science questions around a specific topic), a listing  of significant accomplishments,
and a list of acronyms can be found  as Appendices.
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  Long Term Goal 1:  OPPTS and/or other organizations use the results of ORD's
  research on methods, models, and data as the scientific foundation for:  A)
  prioritization of testing requirements, B) enhanced interpretation of data to improve
  their human health and ecological risk assessments, and C) decisionmaking regarding
  specific individual or classes of pesticides and toxic substances that are of high priority.
Ultimate Outcomes: Research will provide improved methods, models, and data for OPPTS' use in
requiring testing, evaluating data, completing risk assessments, and determining risk management
approaches.  Specifically the research will result in the development by ORD and implementation
by  OPPTS  of more efficient  and effective testing paradigms that  will be better informed  by
predictive tools  (what chemicals, improved targeting cost less, less time, and fewer animals);
improved methods  by which  data  from  the  more efficient and  effective  and existing  testing
paradigms can be integrated  into risk assessments; and that OPPTS uses  the result of  ORD's
multidisciplinary research approaches, that it specifically requests, for near  term decisionmaking
on high priority individual or classes of pesticides and toxic substances.

Synopsis: ORD intramural and extramural research is:
•   developing and applying the latest molecular and computational approaches to produce the next
    series of chemical prioritization tools and toxicity testing approaches;
•   enhancing data interpretation by evaluating the diagnostic value of data obtained from  current
    toxicity testing guidelines in order to develop improved targeted test methods for major  classes
    of pesticides based on defined  modes-of-action  and identification  and characterization  of
    genomic and proteomic biomarkers;
•   characterizing toxicity profiles of perfluoroalkyl chemicals, examining the  potential for selected
    perfluorinated telomers to degrade to perfluoroctanoic acid (PFOA) or its precursors,
•   developing methods and models to forecast the fate of pesticides  and byproducts from  source
    waters through drinking water treatment systems and ultimately to the US population,
•   providing exposure methods for large-scale human studies, and
•   addressing specifically identified research needs by studying chromated copper arsenate-treated
    wood, asbestos, chiral pesticides, lead-based kits.

Protecting human health and the  environment from harmful agents carries  the  challenge  of
developing the capability for assessing hundreds of possible hazardous effects for tens-of-thousands
of important commercial chemicals.  Establishing strategic priorities to focus available laboratory
testing resources on those chemicals posing the greatest potential risks is  essential  to EPA  in
minimizing  environmental risks from  harmful agents.   Over the  last three decades, ORD has
developed for EPA an extensive arsenal  of test methods needed in all aspects of regulatory risk
assessment.   ORD  will  continue,  through the SP2 research program, to refine  many of these
methods and to reduce the uncertainty with respect  to  interpreting  the  results of tests in EPA
decisions.   However,  through the  SP2  research program,  ORD  will also address  the  greater
challenge of developing the science  necessary for EPA to know when and how to apply those test
methods to gain greatest insight into the potential risks of a specific chemical.

Over the years, collaborations  between ORD and OPPTS have resulted in substantial advances in
human health and ecological testing paradigms and improved risk assessment approaches. In order
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to address future challenges to providing credible scientific information, timely and efficiently, to
support risk assessment and risk management decisions for industrial chemicals and pesticides, in
2004, OPPTS and ORD developed a  strategic plan for research in support of OPPTS program
activities. This plan provided a general framework to guide the development and implementation of
new scientific programs and specific advancements that will further effective regulatory decision-
making.  Thus, the long-term solution to meeting this challenge will not be the generation of more
data faster,  but rather determining what  specific  effects data, for which chemicals,  and which
exposures is essential to assess and manage risks appropriately. In this context, the Agency requires
sufficient, targeted, credible information from which to make decisions. Consistent with this view is
the consideration of time and cost efficiencies associated with the generation and interpretation of
toxicity data and the sound and responsible use of animals.

OPPTS is responsible for regulating certain chemicals for which there are little or no toxicological
or exposure data (e.g., Pre-Manufacture Notification (PMN) and High Production Volume (HPV)
chemicals,  inert pesticide  ingredients, antimicrobial  pesticides). Therefore, there is a need for
creating  ways  to accurately predict  the  toxicity  and  levels of exposure  for  these  chemicals.
Predicting the potency, activity, and exposure to these chemicals will enable OPPTS to make better
informed decisions  as to whether or not empirical studies are required  to further refine a risk
assessment  for regulatory decisionmaking.  Current approaches  for  testing chemicals  require
extensive resources.  Therefore, priority setting approaches must be developed  to determine the
sequencing  of  chemicals  or classes  of chemicals to assess for a specified toxicity endpoint.
Additionally, while extensive data sets are generated for many toxicity endpoints currently used in
risk assessment, efficiency can be gained in using targeted testing to reduce critical uncertainty
while minimizing resource utilization. The current inability to estimate endpoints sufficiently to set
hypothesis-driven risk-based priorities is  the result of a lack of understanding of pathways  of
toxicity and how they can be initiated by chemicals, as well as by a lack of methods to model the
complex behavior of chemicals.  By  having an understanding of the initiating events of critical
toxicity pathways OPPTS and ORD will be able to use credible ex vivo techniques to estimate the
toxic potential  of chemicals and  allow them to be ranked/prioritized for their potential to elicit
adverse outcomes.  With the development and application of new computational and molecular
tools, it  is anticipated that in silico and  in  vitro  techniques for prioritization and  screening  of
chemicals for  toxic effects  resulting from  exposure to PMNs,  HPV/inerts  and antimicrobial
chemicals is highly feasible over the next seven years. The  determination of possible levels  of
exposure to these  chemicals will also need to be included  into any  screening or prioritization
program.  Thus, of the issues facing OPPTS, the need to develop more efficient ways to screen and
prioritize chemicals for testing to acquire sufficient, targeted, credible information for decision
making is of high priority.

To overcome these gaps,  and to move toward a  more sustainable risk assessment paradigm  to
support TSCA,  FIFRA, and FQPA decisions, the SP2 research conducted under LTG 1 will provide
EPA with predictive tools for hypothesis-driven prioritization of testing requirements and enhanced
interpretation of exposure, hazard identification,  and dose-response information.   The  research is
complementary to and is coordinated with ORD's Computational Toxicology  (Comp Tox) Research
Program.

In   2002,   EPA   began   a   new   research   program   in   Computational   Toxicology
(www.epa.gov/comptox) to better understand the relationships between sources of environmental
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chemical exposures and adverse outcomes.  Computational toxicology is defined as the integration
of modern computing and information technology with the technology of molecular biology and
chemistry to improve EPA's  prioritization of data requirements and risk assessment  s for toxic
chemicals.  Three strategic objectives of the initiative are to:  1) improve understanding of the
linkages  in the continuum between the source of a chemical in the environment and  adverse
outcomes, 2) provide predictive models for screening and testing, and 3) improve quantitative risk
assessment.  While ORD has a separate Comp Tox program, many other ORD core and problem-
driven research programs are also developing and  applying  comp  tox tools to address their
objectives.  These research activities are coordinated with those of the Comp Tox Research Program
and       are        linked       under        the       Comp       Tox       Framework
(www.epa.gov/comptox/comptox framework.html).  This  is the case  with the  comp  tox related
activities in SP2, where as apparent, the objectives of both Research Programs have commonalities,
especially in the  area of developing/applying methods for the  prioritization  and  screening of
chemicals.  LTG 1 of the SP2 Program not only includes intramural comp  tox research but  also the
extramural  STAR component  of the Comp  Tox  Research Program.   Through issuing targeted
requests for application, ORD is engaging non-profit organizations, to conduct research in areas that
complement our intramural activities.  The STAR Comp Tox program, as noted below, is providing
a unique opportunity for academic  scientists to  work  cooperatively with EPA  scientists on
advancing the development and application of computational and molecular methods.

The research in SP2 LTG 1 also builds upon the screening and  testing  efforts underway in the
Endocrine     Disrupters     (www.epa.gov/osp/myp/edc.pdf)     and      Human     Health
(www.epa.gov/osp/myp/HH%20MYP%20Final.pdf) MYPs by applying  tools, techniques and
knowledge to problem-driven research in support of the major OPPTS needs.

The development of research strategies and MYPs in ORD  serves  to  keep research activities
focused  on  the  frontiers that cause  the greatest uncertainties  in  risk  assessment and risk
management.  In designing the strategic framework for the SP2 research program, it became clear
that the broad spectrum of regulatory responsibilities in OPPTS  presented  special problems for
research  planning. While there was general agreement that ORD should focus more resources on
fewer and more central research needs of the many OPPTS programs, there was also the  concern
over having all available resources committed to longer-term research and not having the flexibility
or resources to respond to special scientific needs that arise. For this reason it, part of LTG 1  is
devoted to addressing these shorter term targeted needs. It is anticipated that as these needs are met,
that they will be replaced with other emerging needs of priority at that future time.  The compilation
of work  described  further under LTG 1 represents the  high priority shorter-term research  being
conducted to address the currently identified targeted needs by  OPPTS. The  shorter-term research
falls under  two separate APGs. Research on the perfluorinated chemicals (PFCs) began several
years  ago focused only on characterizing their potential adverse health effects. More recently ORD
has been asked  to  conduct research on characterizing their  environmental  releases, fate and
transport, and degradation. Therefore, the body of research on the PFCs is large enough to merit a
separate APG.

APG  -  Develop and  validate virtual chemical and  alternative  methods for  risk-based
prioritization and screening of chemicals - FY 2015

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High quality data are essential to the development of predictive models, such as QSARs, needed to
prioritize  chemicals for hypothesis-driven regulatory testing. Where large numbers of chemicals
exist that need to be assessed based on little or no measured data, QSAR predictions can be used to
prioritize which should be tested, when testing should occur, and which endpoints should be tested.

Such approaches are needed to form  the  basis for targeted regulatory  testing to increase  risk
assessment efficiency, but are only possible where toxicity pathways are  well  defined and where
assays are  available for sufficient strategic testing  to build applicable models.  Strategies to
systematically  test  within large chemical  inventories  are included in LTG 1 to minimize the
collection of redundant information and to maximize the understanding of the attributes of chemical
structural that initiate specific biological interactions. Targeted testing approaches in LTG 1 seek to
incorporate new in  vivo protocols that provide data with less uncertainty and/or to provide in vitro
assays that have the potential to develop into high-throughput (HTP) systems when they are linked
to the in vivo endpoint.

Tools needed for screening and prioritization follow a logical  progression from early method/assay
development, through the application of an assay or suites of assays in  a diagnostic manner to
elucidate toxicity pathways, to the systematic testing of multiple chemicals to identify  groups that
initiate toxicity in a common way. Once testing can be done within a pathway, chemical testing can
be  focused to determine  features  of chemical structural  that  facilitate chemical-biological
interactions. Quantifying the structural  requirements through  which a chemical initiates a toxicity
pathway allows the  prediction of probable toxic interactions for a given chemical. As the knowledge
base grows, it  can be used  as  a basis to prioritize testing requirements based on likelihood of
causing effects.

Short and intermediate-term research in this area focuses on providing better  access to existing
models, developing models where toxicity pathways are well understood,  and providing tools and
approaches that can be applied in the longer term as understanding of additional pathways allow.
Long-term research in this area will attempt to integrate data generated from intermediate-term
projects including  in vitro screens and genomic and proteomic approaches with new knowledge of
toxicity pathways to develop predictive models that  specifically  address data gaps  in  the  risk
assessment process. Systematically collected assay  data  in short- and intermediate-term projects
will, using  tools and approaches currently under development, serve as the  basis for toxicity
pathway-based  QSAR prioritization protocols in the future for the  major endpoints of regulatory
concern. The gradual development of a library of pathway-specific models will eventually allow the
prioritization of testing endpoints.

ORD research is:
•   Determining the  chemical structural  requirements for initiation of distinct  toxicity pathways by
    incorporating QSAR-based hypothesis generation, strategic chemical selection for hypothesis testing, in
    vitro assay optimization and targeted testing, and QSAR evaluation and improvement for mechanistic
    classifications for OPP pesticidal inerts and antimicrobials, chemicals for which data  are lacking and
    predictions needed. Products include guidance on development  of QSAR prioritization models for toxic
    effects endpoint in the context  of well-defined toxicity  pathways and demonstrated application of the
    methods for  OPP chemical lists for which testing priorities are  requested. (Project  1.1.1 - refer to more
    detailed description of each project in Appendix IV).
•   Upgrading and expanding the capabilities of ASTER (Assessment Tools for the Evaluation of Risk) to
    rank large lists of chemicals  based on data available in ASTER  (e.g., ecotoxicity, environmental
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    partitioning,  environmental persistence,  and chemical bioconcentration in tissues),  and searching the
    ECOTOX database for structural analogs. This will facilitate the identification of structural analogs and
    associated toxicity information to estimate potential  hazard of untested chemicals  or chemicals with
    limited toxic effects information. 1.1.1
•   Refining an existing metabolism simulator to focus on metabolic transformations most likely to increase
    toxic potential for estrogenicity.  The research helps in better understanding toxicity pathways from
    initiating  event to response for metabolically activated chemicals. A computational tool is provided that
    allows prediction and prioritization of chemicals for which measured data are lacking.  1.1.1
•   Developing  and applying Nuclear Magnetic  Resonance-based metabolomics for improving chemical
    exposure  and risk assessments. ORD  will produce:  1) validated markers of biologically relevant
    chemical  exposure to important classes of pesticides and toxic chemicals, 2) information on the temporal
    and compensatory aspects of chemicals  exposures, 3) information on similarities  and  differences
    xenobiotic metabolism and the impact of exposures across key species (small fish, rats, etc.), and 4)
    information on the linkage of exposure events to whole organism adverse outcomes. 1.1.2 (part of this
    project is  linked to second grant described under 1.2.4 and 2.1.3)
•   Developing  in vitro cell culture models of the key events in brain development and methods to measure
    behavioral, morphological and  neurochemical outcomes in a limited number of non-mammalian species.
    The database that results from  data generated by testing the set of known developmental neurotoxicants
    using HTP methods will  provide OPPTS with information on the utility and limits of the  screening
    battery, and  provide guidance  for  the interpretation  and  potential use  of data  from these  alternative
    methods in a risk assessment context. 1.1.3
•   Utilizing  Sertoli cell cultures challenged  with a panel  of known reproductive toxicants to identify insult
    induced effects in vitro. Emphasis will be on identifying markers with the potential to be used in current
    one generation tier testing and research could result  in the development of a HTP  cell culture  based
    screen for assessing testicular and epididymal insults.   1.1.3
•   Examining protein expression  profiling  as a means  to screen chemicals for  their mode of action for
    pathway  specific toxicity.  Protein  profiling  is also  being investigated to develop  an "omics"-based
    approach  to understanding differences in  species sensitivity to chemical categories, and incorporated into
    existing in vitro and short-term in vivo assays needed to support hypothesis-based risk assessment and
    regulatory decisions. 1.1.3

In 2002, a request for applications (RFA) of proposals from non-profit organizations was issued
through the  Comp Tox STAR Research  Program on new approaches to the development of HTP
screening systems for identifying chemicals with  estrogen, androgen, or thyroid hormone activities.
In its initial  phase, the Comp  Tox  Research Program used  the endocrine system as a 'proof of
concept.'  The  research awards  made through this solicitation  are expected to contribute to the
development of HTP screening systems to assist in prioritization of chemicals for  further screening
and testing of their potential as endocrine disrupters.  The methods generated from these studies will
provide for multiple platforms for the HTP screening of potential endocrine disrupting chemicals, in
some cases even  allowing for remote, near real-time monitoring of potential  endocrine disrupting
chemicals in the environment.  Under this program, ORD is supporting extramural research that is:
•   Developing  and applying a bioluminescent yeast-reporter system for screening chemicals for estrogenic
    and androgenic effects.  1.1.4
•   Developing  a mechanism-based, high-throughput screening assay for evaluating estrogen, androgen, and
    thyroid (EAT)-like activities in an  invertebrate species that also  can  be used to evaluate  interactive
    effects of endocrine-active compounds through receptor cross-talk.  1.1.4
•   Investigating the proposition that perturbations in the  normal amount or timing of a hormone-regulated
    gene product can be taken as evidence of chemical exposure and used as  an endpoint in a screening assay
    using the  zebrafish to detect potential endocrine disrupting activity.  1.1.4

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•  Developing a rapid, sensitive, biologically-integrated screening assay to identify endocrine-disrupting
   chemicals (EDCs) using a medaka fish model. 1.1.4

APG - Evaluate  and  provide guidance regarding the sensitivity and predictive value of
current  test methods  and  those under development  for  the improved  identification  and
characterization of the potential of environmental chemicals to cause human health  and
ecological risks - FY 2013

SP2 research is developing hypothesis-driven risk assessment paradigm that moves in a logical and
transparent  manner  from  a  process that requires  extensive toxicity testing followed  by the
elimination  of information not relevant to the  assessment for all  chemicals (e.g., food-use
pesticides)  to a paradigm where far fewer tests are needed but the  experiments are designed to
elucidate the toxicity pathways that are triggered by a given chemical/class of chemicals to identify
what additional specific in vivo information will be most relevant to the assessment. OPPTS could
apply  such  a  paradigm to both new and  existing chemicals  including  conventional food-use
pesticides. The goal is to be able to continually make scientifically sound regulatory decisions but
with significantly less required data  generation. This process could be applied to chemicals from
across OPPTS programs because the  amount of data required to understand toxicity pathways from
initiating events to adverse outcomes will be limited only to endpoints of interest which will greatly
limit the number of in vivo tests needed to determine a chemical's  toxic potency.

SP2 research on database development projects are making the most  efficient use of existing OPP
registrant data submitted under a wide variety of testing guidelines.  The large quantity of guideline
study data has the potential to advance the understanding of toxicity pathways by  providing in vivo
outcome  information collected under standardized guidelines.  However, to reach this potential the
data  need to be widely  accessible and searchable. LTG 1  research is  addressing these needs for
toxicity endpoints, and  chemical metabolism and  degradation pathways. Outputs of the database
efforts will contribute both to enhancing data interpretation and to prioritization  and screening.  The
first  step of the approach is  to identify where  data already exist and  to make them more easily
accessible for evaluation by both risk assessors and researchers. Thus, building and populating well-
designed databases is  a near-term emphasis because it  is the key to  making  progress in the
intermediate and longer-term. Searchable databases contribute in multiple ways to LTG 1 projects
by:
•  Greatly  assisting data  interpretation  by  allowing  efficient  access to  information otherwise
   unevaluated due to inaccessibility or simply not knowing it exists
•  Allowing records to be grouped by user-defined descriptors  to facilitate evaluation of data in
   new ways and to discover associations previously not examined
•  Providing better  access to program  office data to allow  identification of  critical linkages in
   toxicity  pathways thus enhancing interpretability  of  in vitro and biomarker data linked to
   adverse outcomes
•  Assessing where knowledge gaps exist, and
•  Allow examination of where correlations can and cannot be made across  endpoint measures,
   across chemicals, and across species

ORD is working with OPPTS risk assessors and  others to design databases that will:
•  Allow better understanding of existing data
•  Allow systematic evaluations of some of the data resulting from newer guidelines and protocols
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   that have as yet not been evaluated
•  Facilitate comparisons of information available in current tests with newly developed tests to
   guide the development of new testing approaches in a hypothesis-driven manner to target where
   efficiencies or enhanced interpretation can be gained.

Where  a toxicity pathway is  sufficiently understood, and where  assays are available for that
pathway, systematically harnessing the existing knowledge-base needed  may  provide  all  that is
needed for the development of more realistic scientific basis to assess risk. In that vein, toxicity
pathway-specific QSARs for prioritization and screening are under development for currently well-
defined pathways building on the OPP historical database.  In the mid- and longer- term research is
underway to demonstrate the approach and build the tools needed which can be applied in the future
as more key pathways are elucidated.

Efficient access to existing data to identify similar acting chemicals based on toxicological outcome
can be achieved by developing searchable databases. Collaborative efforts  between ORD and OPP
risk  assessors  under LTG  1  will focus on outcomes  that  are  of high priority to enhance
interpretation of data and that optimally leverage existing ORD  research approaches and expertise,
so that both risk assessors and researchers maximally  benefit from these efforts.  LTG 1 database
projects  also support the  need to  enhance  our current  ability to  interpret many  types of data
submitted under existing guidelines. In some instances sufficient data have only recently become
available as relatively  new  guidelines were introduced  over  the last several years.   Areas are
identified where extensive evaluations of newer datasets and examination of test protocols by ORD
and OPPTS collaborators will be undertaken to help inform current risk evaluations and to direct the
development  of  hypothesis-driven  predictive,  diagnostic  markers   (biochemical,  proteomic,
genomic) for given risk  endpoints  (e.g., immunotoxicity, neurotoxicity, reproductive effects).
Hypothesis-driven predictive, diagnostic markers will help us learn if we use the results to form the
basis for further targeted testing. The goal is an increased understanding  of toxicity pathways to
guide testing and to generate high-quality datasets.

Where toxicity pathways are not sufficiently understood  (e.g.,  where there are critical knowledge
gaps in the continuum from chemical initiation of the toxicological process to its manifestation as a
whole organism adverse outcome) research will be undertaken in concert with ORD's core research
programs in Human Health, Endocrine Disruption and Computational Toxicology to assess current
understanding of the events leading to adverse effects, to identify critical  knowledge gaps,  to
understand commonalities in toxic pathways to  develop integrative new methods that provide
information on multiple effects, and to develop in vitro assay approaches for rapid screening.

Under database development, ORD  researchers are collaborating with OPPTS scientists to:
•  Design a database to provide access to currently non-accessible metabolic map information that exists in
   OPP files. Access to metabolism data in a searchable format is key to understanding the role of metabolic
   activation in toxicity pathways, and to being able to generating targeted hypothesis that will address the
   highest priority uncertainties for the types of chemicals of most concern to the program offices. 1.2.1
•  Design a chemical degradate pathway database to provide ready access to pathways of degradation
   associated with identified reaction types, for specified bioassay  conditions across all study types and
   chemicals of concern. The development of searchable databases is  key to efficient use of existing
   information and moving toward a  new paradigm based on hypothesis-driven testing and prioritization.
   1.2.1
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•   Build the template to incorporate developmental neurotoxicity data from the literature and Office of
    Pesticide Programs (OPP) datasets into an inclusive searchable database.  The database  developed
    could  serve  as  a model  for  similar databases  for other endpoints (e.g.,  reproductive toxicity,
    immunotoxicity). 1.1.3

In conducting research to enhance interpretation of existing guideline data and those from new
methods, ORD researchers are:
•   Evaluating and  providing guidance on  major issues regarding the  methods used  in the current
    Developmental Neurotoxicity  (DNT)  Testing Guidelines.  Project outputs will include guidance
    documents for data requirements, standardized protocols and data evaluation, for use both within and
    outside the Agency which should be  of immediate use to OPPTS in terms of data interpretation,
    requirements for future testing, and guidance for test method development and refinement. 1.2.2
•   Identifying sensitive in  vivo and in vitro approaches for recognizing and screening alterations of
    rodent immune function through the application of genomics.  A comparison of the sensitivity of in
    vivo and in vitro immune function results versus genomics should lead to the identification of the
    most sensitive metric(s) for immunotoxicity testing that could be incorporated into a  single assay to
    assess  for potential neurotoxic and reproductive effects and could lead to a more efficient use of
    animals in testing. 1.2.3
•   Using  in vitro  and in  vivo approaches to understand and  discriminate  the  compensatory and
    toxicological responses  of  the highly  regulated  hypothalamic-pituitary-thyroid (HPT)  system.
    Changes in gene expression in the pituitary, thyroid, and peripheral tissues under normal  conditions
    and following exposure to chemicals known to interfere with  TH synthesis will be linked with
    functional measurements of key hormones  and enzymes that are part of the HPT pathway,  all of
    which will be interpreted in the context of organismal-level effects. 1.2.3

In 2003, an RFA was released on the use of systems biology approach in hazard  identification
and risk  assessment.   Applicants  were asked to develop, with  or without cross  species
extrapolation, integrative, quantitative  models of the function  of the HPG or HPT axes with
emphasis  on  the  descriptions  of the normal  physiological  processes and mechanisms  of
perturbation following exposure to endocrine disrupting chemicals  in  routinely used  animal
models. Through this RFA, ORD is supporting research that is:
•   Developing a screening method to use molecular techniques to screen for effects of chemicals on the
    hypothalamic-pituitary-gonadal (HPG) axis with a special emphasis on steroidogenic pathways and
    hormonal control mechanisms along the HPG-axis in the Japanese medaka.  1.2.4
•   Developing a computational model to evaluate  molecular and protein biomarkers  in relation to
    reproductive  dysfunction in  fathead minnows exposed  to environmental estrogens.  The model is
    incorporating a number of biochemical  endpoints along  the entire HPG  axis,  directly  evaluating
    physiological changes and reproductive endpoints and the pharmacodynamics and kinetic distribution
    of the contaminants. Awardees and ORD scientists are working collaboratively on this project (see
    complementary research under 2.1.3) through cooperative agreements.  1.2.4
•   Developing a computational model  that will identify and  predict critical estrogenic  endocrine
    disrupter elicited changes  in gene  expression  which play  a central  role  in  the observed
    physiological/toxic effects based on systematic and quantitative data  obtained from comparative in
    silico, genomic, molecular and histopathological approaches using the rat uterus.  1.2.4

In 2004, an RFA was issued to establish an environmental bioinformatics research center.  Two
awards were made as cooperative agreements to enable close collaborations with scientists from


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ORD and EPA program offices.  The research is expected to contribute to development and
application  of dose-response information  analysis and enhance current quantitative  risk
assessment practices and reduce uncertainties.  The computational methods developed will link
to data from molecular  toxicology and other studies in order to  move risk assessment from a
hypothesis-driven science toward a predictive science. ORD is supporting research that is:
•  Developing novel analytic  and  computational methods,  creating efficient user-friendly tools to
   disseminate the methods to the wider community, and applying the computational methods to data
   from  molecular  toxicology and  other  studies by establishing  the  Carolina  Environmental
   Bioinformatics Research Center.  1.2.5
•  Addressing, in a  systematic and integrative manner,  multiple elements of the toxicant Source-to-
   Outcome sequence as well  as developing cheminformatics tools  for toxicant characterization by
   establishing the New Jersey Research Center for Environmental Bioinformatics and Computational
   Toxicology.  1.2.5

In late 2006, an RFA will be issued to solicit proposals with the goal of developing predictive
environmental and biomedical computer-based simulations and models that address data gaps in
environmental and human health  risk assessment and will strengthen the ability of predictive
scientific data to guide sound future scientific policy and decisions. 1.2.6

APG -  Develop the scientific underpinnings  related  to the  effects, exposures, and risk
management of perfluorinated chemicals to inform Agency risk assessment/management
decisions - FY 2013

A number of PFCs are increasingly being  used for  a  variety of household  and industrial
applications.  These include the surfactant coatings for fabrics and paper products, fire-fighting
foams,  non-stick cookware,  electronic  etching baths,  and  insecticides.    EPA  began  an
investigation of some of these chemicals because perfluorooctanoic acid (PFOA) is found at low
levels both in the environment and in the blood of the general US population, is very persistent in
the environment, and causes developmental and other  adverse health effects  in  laboratory
animals.  EPA has  summarized its concerns and identified data gaps and uncertainties about
PFOA in  a notice  published  in  the Federal  Register (http://www.epa.gov/fedrgstr/EPA-
TOX/2003/April/Dav-16/19418.htm. OPPT-2003-0012).

EPA has negotiated with multiple members  of the regulated community to  develop data on
PFOA through enforceable consent agreements, memoranda  of  understanding,  and voluntary
commitments.  Information on these activities can be obtained through EPA's PFOA website
(www.epa.gov/oppt/pfoa). One of these compounds, perfluorooctyl sulfonate (PFOS) has been
withdrawn  from the commercial market by the 3M company in 2000, but is still readily available
from other  overseas manufacturers. At present, little information is available concerning sources
of environmental exposures,  environmental fate and transport,  and potential adverse health
effects of these PFCs. Completing PFOA risk assessment actions is one of the Program Priorities
of       the        OPPTS        Assistant       Administrator       in        FY06-07
(www.epa.gov/oppts/pubs/programpriorities.htm).  While some data are being developed by the
regulated community, there are other scientific needs that have  been identified by OPPTS as
critical to their being able to complete their risk assessments for the PFCs.


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ORD research is, therefore, focusing on these unmet needs and is:
•   Characterizing the developmental toxicity of PFOS and PFOA, determining to what extent data on
    these chemicals can be predictive of effects to other environmentally relevant PFCs, and identifying
    an alternative animal model to  the rat.   This research addresses OPPTS's immediate need to
    characterize the hazards of these chemicals for human populations  and will help form the basis of
    other projects that focus on modes of action (MOAs) for PFCs toxicity.  1.3.1
•   Elaborating on the already known pharmacokinetic profiles of PFOS  and PFOA in laboratory rats by:
    1) studying profiles in the mouse; 2)  discerning differences between immature and adult rodents; and
    3) evaluating the  patterns tissue accumulation.  The  studies will be conducted in concert  with  a
    comprehensive investigation coordinated by NTP, OPPTS  and ORD  of 18 PFCs identified by OPPT.
    1.3.2
•   Evaluating the immunotoxicity, hepatotoxicity and the endocrine disruptive (thyroid and estrogen)
    potentials of PFCs and determining  to what extent potential effects seen in rodent animals  can be
    extrapolated to humans.  Studies will also be conducted to assess the MOA of any adverse effects and
    whether developing animals are more sensitive to some of the effects than adults.  1.3.3
•   Characterizing the underlying MOA responsible for the adverse effects  caused by PFCs, including
    determining the involvement of peroxisome proliferator activated receptor (PPAR) signaling pathway
    in PFC toxicity. Results will provide information regarding the affinity of PFCs for PPAR isoforms,
    and comparisons of the affinities of the compounds in rodent versus human receptors. 1.3.4
•   Developing  sampling and analytical  methods to characterize the distributions of PFOS, PFOA, and
    other  PFCs  in  key  environmental  and biological matrices that are important for assessing
    environmental transport and fate and human exposure.  1.3.5
•   Developing data on the stability and distribution of selected perfluorinated organic chemicals in real-
    world environments.  Research is being performed to assess the potential for fluorotelomer-based
    polymer products  (FBPPs) to degrade in a variety of  soil  and sediments  and characterize the
    environmental distribution of perfluorinated compounds in  soils. 1.3.6
•   Determining  fluorotelomer alcohol  (FTOH) polymer product stability during wastewater  (WW)
    treatment by:  1)  developing  analytical  and  experimental  methods  to characterize fluorinated
    surfactants in various environmental matrices, 2) describing  the composition  of FTOH polymer
    formulations released  to the  environment via  down-the-drain  disposal,  3) determining  the
    environmental loadings of FTOHs and PFCs from WW treatment, and 4) determining  the potential
    for FTOH polymer products to transform or degrade during WW treatment.  1.3.7
•   Characterizing the source, transport,  and fate of PFCs in the indoor environment and the factors that
    may affect PFCs release from articles of commerce (AOC) by  determining the PFCs content  in new
    AOC, characterizing the PFC emissions from aged AOC containing fluoropolymers or fluorotelomers
    by chamber studies, and identifying the major indoor exposure routes for the general US population.
    The findings will suggest potential risk management solutions. 1.3.8

APG -  Develop  the scientific underpinning  related  to  the effects, exposures, and  risk
management of specific individual or classes  of pesticides  and  toxic substances  that  are of
high priority to the Agency to inform Agency risk assessment/management decisions - FY
2010

An area  of critical concern to OPPTS and the Office of Water is the ultimate product that results
and is available for exposure to the environment and public when  a pesticide or toxic chemical
enters a  drinking water  distribution system.  Drinking water (DW) treatment often has a large


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effect on pesticides and toxic chemicals that occur in source waters; and both OPP and OPPT
have  articulated  their high  priority need to incorporate these effects  into  chemical  risk
assessments. FQPA requires OPP to consider all anticipated dietary exposures when conducting
risk assessments for pesticides.  DW is considered a potential route of exposure because many
DW sources contain detectable  levels of pesticides  and toxics.   The  majority  of the US
population consumes treated DW.  It is anticipated that DW treatment will: a) partially remove
some chemicals, b) transform some chemicals (perhaps to more-toxic byproducts), and c) have
virtually no effect on some chemicals.  Unfortunately, very little data has been collected, and
very little mechanistic information is available, on the effects of DW treatment processes on
pesticides or on any other chemicals.

OPP has announced a policy  to systematically consider DW treatment effects on pesticides for
FQPA risk assessments.  For this policy to make risk assessments significantly more certain,
pesticide-specific information on  DW treatment  effects is  needed.   Also, reliable tools to
extrapolate effects  across  chemical  classes are  required.   These  needs,  which  have  been
articulated by OPP, are driving this  research.  Without the type of information and tools that
ORD will provide, OPP will  have to assume that DW treatment has no effect on a pesticide or
toxic chemical. This may be overly- or under-conservative, depending on the pesticide or toxic
chemical, and the treatment conditions. ORD has been partnering extensively with OPP and OW
on this research.  Addressing DW and pesticides interface issues is one of the Program Priorities
for       the       OPPTS        Assistant        Administrator        in        FY06-07
(www.epa.gov/oppts/pubs/programpriorities.htm).  To address these needs, ORD research is:
•  Producing tools to  address OPP's DW treatment research needs by:  1) developing/evaluating a
   protocol, 2) performing DW treatment studies, 3) providing chemical-specific information on the
   effects of water  treatment  on pesticide transformation pathways,  4) providing  physicochemical
   parameters for transformation products, and 5) developing predictive models for forecasting treatment
   effects. This research will allow OPP to provide a protocol to pesticide manufacturers for their use in
   submitting data in a consistent manner and will provide OPP with DW treatment data and guidance
   on their interpretation for inclusion in their risk assessments.  1.4.1

Chromated copper arsenate (CCA) is a chemical wood preservative injected under high pressures
to protect wood from decay and insect damage.  In February  2002, the manufacturers of CCA-
treated  wood asked EPA  to  remove the  registration of  CCA for residential  use  including
playground equipment, decks, and landscape timbers.   The CCA-treated  wood manufacturers
phased out production and use of CCA in favor of alternative  chemicals effective December 31,
2003 (www.epa.gov/oppad001/reregistration/cca).  Of current remaining concern is the possible
dermal exposure of children to the arsenic contained within the CCA-treated wood that has been
used for  playground  equipment,  decks,  and outdoor  furniture.  Since there  was  little  data
available  on  efficacy of sealants  and coatings to minimize dermal  exposure,  ORD and the
Consumer Product  Safety  Commission  (CPSC)  conducted  studies to identify  and evaluate
commercial coatings that can reduce or prevent exposure to arsenic from CCA treated wood.
 To address these  needs, ORD research is:
•  Evaluating the  ability of coatings available to consumers to reduce  dislodgeable CCA residues on the
   surfaces of CCA-treated wood and demonstrating a methodology that industry can use to further
   develop, improve, and demonstrate product performance.  1.4.2


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For the last 10 years,  ORD has been  a part of a  large multifaceted interagency study, the
Agriculture Health Study (AHS), to evaluate pesticide exposures and their potential to cause
adverse health effects in pesticides applicators and their families. The ORD-led AHS Pesticide
Exposure Study was an exposure-measurement field  study for a subset of agricultural pesticide
applicators and  participating family members in  the larger  AHS cohort.   It was conducted to
provide information to assess and refine the exposure-classification procedures developed from
the AHS  questionnaire data and for the National Cancer Institute (NCI)  and NIEHS to better
understand factors affecting pesticide  exposures for  agricultural pesticide applicators and their
families.  ORD's contribution to this study is:
•  Generating high quality exposure data for classifying pesticide exposures for agricultural applicators
   and assessing and refining the NCI/AHS exposure algorithms. Identifying  and understanding key
   exposure factors can guide development of improved exposure reduction strategies and guidance
   developed by OPP and other organizations and increase the value of epidemiological study results in
   pesticide exposure and health assessments. 1.4.3

Through its existing chemicals program, OPPTS  oversees  the implementation of its asbestos
program (www.epa.gov/asbestos).   Asbestos is commonly used as an acoustic  insulator, in
thermal insulation, fire proofing and in other building materials. While much is known about the
effects  of and  exposures to asbestos, there  are  still some remaining  research needs.   ORD
research is:
•  Providing scientifically sound sampling and analytical approaches for measuring asbestos fibers in
   various media and applying these measurements to  assessments of human  exposures to asbestos
   fibers. The research focuses on two key areas:  1) determining which types of measurements are
   needed to support asbestos exposure assessments in likely real-world scenarios, and 2) comparing the
   efficiencies and  effectiveness of available  asbestos  sampling and analytical techniques used in
   exposure assessments.  1.4.4
•   1)  Evaluating aerosolization of asbestos  and  related fibers from  bulk materials  to develop  a
   framework for modeling asbestos breathing zone concentrations generated by activities of varying
   intensity on outdoor and indoor surfaces. 2) Obtaining asbestos fiber releasability data from soil and
   carpet for calculation of emission factors. These data and model(s) will allow EPA Regional  Offices
   and others to make rapid decisions about whether a soil or other bulk material is contaminated with
   asbestos. 1.4.5

On January  10, 2006, EPA proposed  new  requirements  in the Federal  Register to  reduce
exposure to lead hazards created by renovation, repair, and painting activities that disturb lead-
based  paint (www.epa.gov/lead).  The  proposal  supports the  attainment  of  the Federal
government's goal  of eliminating childhood  lead  poisoning by 2010.  Implementing targeted
actions to meet the 2010 lead  goals is one of the Program  Priorities for the  OPPTS Assistant
Administrator in FY06-07  (www.epa.gov/oppts/pubs/programpriorities.htm).   The  research to
date has shown that commercially-available paint lead  test  kits are not an effective means of
identifying  homes  that  do  not contain regulated lead-based paint. To  meet the new rule
requirements, it is necessary that new efficient, cost  effective technologies meeting  the desired
sensitivity within a specified range of false positive and false negatives rates be developed and
made available commercially. ORD research is:

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•  Investigating available state-of-the-art analytical technology for its potential to be modified to meet
   OPPT's needs for the new lead rule. 1.4.6

The  fact  that over  25% of modern pesticides  are chiral  molecules provides an  excellent
opportunity for production of safer products and reduction of the load of unnecessary chemicals
entering the environment.  ORD research is:
•  Addressing the importance of enantiomer selectivity in the fate, persistence, exposure, effects and risk
   assessment of chiral pesticides and other pollutants.  This research provides data for OPP to use to
   better inform risk managers on the  certainty/uncertainty and need for additional  information  for
   pesticides that are either single, enriched or racemic compounds. 1.4.7
   Long Term Goal 2:  OPPTS and/or other organizations use the results of ORD's
   research as the scientific foundation for probabilistic risk assessments to protect natural
   populations of birds, fish, other wildlife, and non-target plants.
Ultimate Outcomes: Results of this research will help the Agency meet the long term goal of
developing scientifically valid approaches to extrapolate across species, biological endpoints
and exposure scenarios of concern, and to assess spatially-explicit, population-level risks to
wildlife populations and non-target plants and plant communities from pesticides, toxic
chemicals and multiple stressors while advancing the development of probabilistic risk
assessment.

Synopsis:  Intramural research is:
•   creating the scientific foundation for conducting probabilistic risk assessments for fish and
    wildlife populations and plant communities by developing: methods for extrapolation among
    species  and  exposure   scenarios   of concern;  models for  characterizing  environmental
    exposures and population biology in spatially-explicit habitats; models to assess relative risk
    of stressors; and tools to define geographical regions/ spatial scales for risk assessment.

OPP is leading the way in expanding ecological risk assessments (ERAs) to provide probabilistic
expressions of risk to aquatic  and terrestrial wildlife populations and  plant communities,
including reducing uncertainties in all  tiers of the risk assessment process as uncertainties that
are extrapolated from limited data sets  are better defined and put into context. For this purpose,
methods are required to support population-level ERAs of increasing degrees of specificity,
detail and realism; to determine the absolute /or relative (incremental) risk of chemical and non-
chemical stressors; and at varying geographical regions/ or other areas of regulatory concern.

The research conducted under LTG 2 of the SP2 MYP is developing efficient methods, including
models, for OPP  to  review, register, and regulate thousands of chemicals in a timely fashion.
OPP's  strategic  direction is  moving  toward  probabilistic  assessments  in response to
recommendations  from  their  Scientific  Advisory  Panel.  ORD has  developed the Wildlife
Research                                  Strategy                                 (WRS,
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www.epa.gov/nheerl/publications/files/wildlife_research_strategy_2_2_05.pdf)  which  describes
a tiered approach using a series of wildlife risk assessments.  A similar tiered  approach is used
with plant risk assessments.  In addition, because neither stressors nor wildlife populations or
plant communities  are distributed  uniformly within the environment, the interplay between
spatial and temporal heterogeneity in wildlife population and plant community structure and
spatial and temporal patterns of stressors is a major factor controlling the severity of effects on
wildlife populations and plant communities.   Thus, a critical feature of  this research is the
development of probabilistic models that deal explicitly with the spatial distribution of wildlife
populations, plant communities and stressors over time. While steps provide general guidelines
for wildlife population and plant community-level risk assessment, the level  of accuracy and
realism appropriate for each step  varies with assessment needs and  management goals.  For
example when applied in  the  context of site-specific risk assessment,  these models  can be
applied to real landscapes, by  interfacing with geographical information systems (GIS).  For
more generalized regional or  national-level  assessments  (e.g.,   for pesticide registration),
simulated (or constructed) landscapes can be used that mimic the general characteristics of the
ecosystems of concern.

Data needs  for each assessment differ depending upon the goals of the  assessment and the
desired level  of  confidence in the  outcome.  For example, screening  level  or "lower tier"
assessments may  involve a relatively simple  evaluation of chemical fate and effects,  including
qualitative judgments about the likelihood of exposure and potential for bioaccumulation.  In the
case of compounds regulated under the PMN process, this  may involve expert opinion about
which class the chemical belongs to and selection of an appropriate analog or QSAR.  A similar
approach may be  applicable to some "inert" ingredients of pesticide formulations, antimicrobials
and HPV chemicals. More involved assessments are required for "active"  compounds covered
by FIFRA as well as some TSCA chemicals of special concern.  These "higher tier" assessments
are more likely to include quantitative evaluations  of effects  and exposure information and may
involve model-based efforts to extrapolate data from surrogate species and chemicals. Currently,
data available for these needs also differ considerably. Registrants of chemicals with  pesticidal
activity are required to provide a core set of data regarding  toxicity to wildlife and vegetation.
In contrast,  under the  TSCA  Section 5 PMN review process, importers or manufacturers of
chemicals are not required to submit any effects data; that is, the EPA can  request toxicity test
results,  e.g., using mammalian (usually in support of human health questions), avian or aquatic
species only if there is reason to believe that a specific adverse effect can occur.

To address these  needs, LTG 2 research is directed toward improving current risk assessment
processes used by OPP and OPPT for new and existing pesticide and chemical risk assessments,
with specific focus  on their needs to:  1) make better use of the current  data that they  receive
during product registration or for existing substance review, and 2) move  the science forward
through the inclusion of more sophisticated data and analyses.

To address the needs of OPP, OPPT and other program offices, ORD's WRS  describes critical
research that is providing the scientific foundation for probabilistic risk assessments to inform
decisions  related to  protection  of natural  populations of  birds, fish and other  wildlife.
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Specifically,  this research focuses on the development of those approaches and tools whose
advancement is most  needed to conduct  spatially-explicit,  population-level risk assessments.
Similar research is necessary to protect native plant populations.  The research conducted under
LTG 2 focuses on three of the four critical steps identified  in the WRS, which must integrate
with the first step (exposure characterization) to complete this process. These three steps include
the development of methods to improve the characterization of effects of chemical and non-
chemical stressors on the fitness of individuals of various species, on the viability of populations
of species with varying  life histories,  and  on  the  dynamics  of spatially-structured wildlife
populations and plant communities inhabiting heterogeneous landscapes.

The  goal of the LTG 2 research is to develop scientifically  valid approaches to assess risks to
wildlife populations and plant communities from multiple chemical and non-chemical stressors.
This requires a  means of  mathematically  integrating dose-response and habitat  suitability
relationships as  well   as  computer platform for  site-specific,  spatially-explicit  population
modeling.  To address these needs, the research is arrayed under the following four APGs.

APG - Provide methods for extrapolating toxicological data  across wildlife  species, media,
and  individual-level response endpoints - FY 2013

Research is emphasizing approaches for extrapolating toxicity data to a broader array  of species,
environmental media, and response endpoints - in particular, the endpoints required  as input to
population response models. Research is continuing to refine the ECOTOX database, Acute to
Chronic Estimation  (ACE),  and Interspecies  Correlation Estimations (ICE) programs. Several
areas within the model have been identified for further development including:  1) identification
and  expansion  of appropriate surrogate species;  2)  increased  representation  of non-pesticide
industrial   organic   and inorganic  chemicals;  3)  increased  representation  of metals;  4)
improvement in taxonomic classification schemes; and 5) model  expansion and modification
integrating novel (example: genomic/proteomic) endpoints.

ORD research is:
•  Reducing uncertainties associated with the current use of (eco) toxicity  data and existing  empirically
   based interspecies extrapolation models to better estimate the toxic effects of chemical exposures on
   wildlife and aquatic species.  This  is being done using two general approaches:  (1) improving
   accessibility and usefulness of available  toxicity information through the  ECOTOX  database, a
   comprehensive Web-based system maintained by ORD; and (2) refining empirical models that  use
   available information to predict toxicity across species (i.e., ICE) and across endpoints (ACE). 2.1.1
•  Furthering the development of mechanistically based models to extrapolate toxicity  information
   among  chemicals, species and  lifestages.  This  is being done using  two general approaches:
   development and testing of a physiologically based toxicokinetic (PBTK) model for a fish  species  and
   an  improved experimental method to parameterize such models;  and, development and  testing of a
   MOA model to predict inter-species differences from in vitro data.  2.1.2
•  Developing  techniques  for extrapolation of toxicological effects  across  endpoints, species  and
   chemicals by utilizing three small fish species, the lapanese medaka, zebrafish and fathead minnow
   and a systems-based approach to define toxicity pathways for model  chemicals with well-defined
   MOA within the HPG axis. The studies employ a combination of state-of-the-art molecular biology,
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    bioinformatic and modeling approaches, in conjunction with whole animal testing protocols.  2.1.3
    (linked to second grant described in 1.2.4)

APG - Provide methods for characterizing population-level risks of toxic chemicals to aquatic life
and wildlife - FY 2015

OPP is redesigning their ecological risk assessment tools to make their risk assessments more
ecologically relevant. In support of this need, OPP and ORD scientists are working to implement
the approaches identified in the Wildlife Research Strategy. This specific effort is aimed at
incorporating population models into the OPP risk assessment process and, thereby, taking a
significant step forward in terms of making their risk assessment process more ecologically
relevant.

ORD research is:
•   Refining population models that have been developed for OPP and integrating these models with the
    exposure and effects models currently used by OPP. Some of the factors for consideration include the
    software code, and how to make a seamless  connection between our population models and OPP's
    existing risk assessment  models; translating OPP effects endpoints into stressor response models that
    can be used in the population models; data quality for population model parameter estimates; model
    interpretation, including  decisions on which population model endpoints will are most appropriate for
    assessing risks; and some level of training and continuing technical support. This effort moves from
    conceptual models of addressing population  response to actual implementation of population level
    analyses. Furthermore, this research advances our understanding of the  usefulness of population
    models for accurately characterizing risks in ecological risk assessments, and determining if the
    quality of the population model is sufficient to answer the risk management question. 2.2.1
•   Providing methods to support probabilistic ecological risk assessments and more explicitly addressing
    higher tier ecological risk assessment needs to take into account greater realism and complexity  in
    projecting population responses to stressors. This is being done using a combination of theoretically
    and empirically based  approaches and field-collected,  laboratory-derived,  and simulation-based
    information. Selected species will include those used frequently in toxicity testing, which historically
    have  provided important information to regulatory process, including the  registration of pesticides.
    Research is addressing  four specific areas:   (1) Probabilistic models, (2) density dependence, (3)
    genetic, and (4) spatial effects. This research will provide approaches and guidance on the need for
    and uses of more complex population modeling approaches as tools for integrating  and  projecting
    more realistic effects of stressors on wildlife populations. 2.2.2

APG - Provide approaches for evaluating the relative risks from chemical and  nonchemical
stressors on spatially  structured wildlife  populations  across large  areas or  regions, and
provide  methods for characterizing population-level risks  of toxic chemical to aquatic life
and wildlife - FY 2009

This research area introduces issues associated with the spatial and temporal heterogeneity  of
populations and stressors, and  extends the analyses under the previous  areas  to applications  in
real landscapes.  Because different stressors tend to be distributed differently in the landscape,
this approach  will address  the interactive  effects  of contaminants,  habitat alteration,  and
introduced  species on wildlife populations.   Models and analyses are being designed both  to
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assess risks from multiple stressors and  to  evaluate the  relative  effectiveness of alternative
management strategies.  The basic GIS and population modeling platform is in development
under the Ecosystem Protection Research Program so as to serve the needs of all EPA Program
and Regional Offices.  Under the SP2 research program, efforts are focused on the relative risk
assessments for bird populations exposed to toxic chemicals and landscape disturbances within
the natural range of the  species. Birds are the class of vertebrate wildlife of principal concern
due to their higher susceptibility as compared to mammals to past and current generations of
pesticide  chemicals, their high visibility,  and high public  concern.   Programs  such  as the
Breeding Bird Survey conducted by the USGS and the multisector!al program Partners in Flight
provide a high level of public input and scrutiny to monitoring bird populations and biodiversity.
ORD is also developing spatially explicit fish population models to assess large-scale pesticide
exposure and risk in coastal areas, significantly enhancing the understanding of OPP staff about
how coastal fish are impacted by pesticides and other stressors.

ORD research is:
•   Enhancing the PATCH model (Program to Assist in Tracking Critical Habitat), is a spatially explicit,
    individual-based life history simulator that incorporates GIS representations of real or hypothetical
    landscapes, and tying it specifically to pesticide issues. The PATCH model is being used to simulate
    wildlife population responses  to pesticide application within  agricultural landscapes and ORD is
    exploring the incorporation of multiple landscape configurations, wildlife life histories, and  stressor
    regimes. 2.3.1
•   Designing and implementing  a probabilistic exposure analysis  system,  undergirded by a suite of
    sophisticated process-based models of pesticide environmental chemistry and biology,  for direct
    assistance to EPA's regulatory  programs in their mandated pesticide risk assessments.  One objective
    is to provide OPP with improved tools for assessing offsite drift of pesticides, and  expanding the
    capabilities of AgDPJFT and AGDISP to assess near-field pesticide drift  from aerial applications by
    including source term algorithms for ground sprayers, orchard airblast sprayers, and re volatilization.
    2.3.2

APG - Provide  improved methods to assess direct and indirect risks to non-target plant
species and plant communities from pesticide use - FY 2012

There is evidence for a wide variation in response of various plant species to chemical herbicides
(about three orders of magnitude), making it difficult to extrapolate potential risk to non-target,
uncultivated plants from the current suite of 10 plant test species (all agronomic species).  Given
the increasing use of herbicides, the FIFRA Science Advisory Panel recommended restructuring
the Tier II risk assessment process to expand the number of species  tested and the  endpoints
evaluated.  Current testing is  limited to a very  small portion of a plant's life cycle that does not
include reproduction and lacks  information relating to  field tests,   especially in terms  of
determining survival of threatened and/or endangered plant species, preservation of native plant
communities, and for maintenance of productive habitat for wildlife.

The objective of the research is  to develop methods to determine the short and long term effects
of pesticides on non-target plants and plant communities, including impacts on wildlife habitat.
This effort includes:  1) Development of spatial analysis tools, i.e., a web-page based, geographic


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information system (GIS)  platform, to identify geographic areas and types of plants with the
greatest risk for non-target herbicide effects.  This spatially explicit framework also will provide
access to  databases  to  support  OPP's  ecological  risk  assessments.  2) Determination  of
reproductive/developmental responses of plants to chemical herbicides for proposed life-cycle
tests.   3) Development  of methodology for greenhouse  and field-plot  studies to determine
herbicide effects  on native plants and plant communities. Both constructed communities with
mixtures of plant species seeded/planted in various relative densities and proportions, and in situ
native plant communities will be studied in the field. Endpoints will include % cover, biomass,
reproduction to provide information on changes in  species composition and system dynamics.
This information will be used to develop guidelines for revised field tests and for assessments of
impacts of pesticides  on quality of plant communities for wildlife. 4) Development of molecular
indicators  (gene expression  and subsequent protein production)  of whether a plant has  been
affected  by  specific herbicides.   This  information will be used  to  predict the  potential
susceptibility of different plant species to herbicides (especially native  plants and threatened and
endangered plant species).

ORD research is:
•   Determining effects  from  off-target movement of chemical herbicides  on  plants and  plant
    communities through: 1) providing a spatially explicit, geographically  based framework to access
    databases  to select plant species  for testing, and for ecological risk assessments; 2) improving
    phytotoxicity testing guidelines,  focusing  on  terrestrial  plant effects,  with  an emphasis  on
    reproductive effects and native plants;  3) improving ecological testing guidelines to determine plant
    community responses; and  4) evaluating molecular indicators can be used to assess whether a plant
    has been affected by specific herbicides, or whether a species may be susceptible to a  specific
    herbicide.
 Long Term Goal 3:  OPPTS and/or other organizations use the results of ORD's
 biotechnology research as the scientific foundation for decisionmaking related to
 products of biotechnology.
Ultimate Outcomes:   OPPTS will use the results from this research program  to  update its
requirements of registrants of products of biotechnology and to help evaluate data submitted to
them.

Synopsis: Intramural and extramural research is:
•   improving  the evaluation  of potential  ecological  effects  of  biotechnology  products,
    specifically plant incorporated protectants (PIPs), on non-target species; the impact resulting
    from the escape of altered plants to the natural environment and the likelihood  and effects of
    gene transfer; the development of pesticide resistance in the target insect species;  the
    development of risk management approaches; and development of methods to  assess for the
    potential allergenicity of genetically engineered plants.
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As  noted previously,  OPPTS,  in carrying out  its Congressional  mandates, evaluates the
environmental risks posed by pesticides and chemicals to safeguard all Americans, including
children and other vulnerable members of the population, as well as our most threatened species
and ecosystems.  OPP regulates the use of all pesticides in the US  and establishes maximum
levels for pesticide residues, including genetically engineered pesticides.  OPPT regulates the use
of industrial chemicals and certain biotechnology products, such as microorganisms used in the
manufacture of specialty  chemicals and bioremediation  agents.  OPPT  also  implements the
Pollution Prevention Act and, hence, has an interest in biotechnology product stewardship that
would lead to "green" chemicals.  OPPT has an emerging interest in certain transgenic plants for
uses such  as  phytoremediation  and  enhanced wood  production although  OPPT does not
implement regulatory oversight in this area at this time.   These new products are  often on the
cutting edge of science and regulatory policy and research is needed to ensure that their  safety
can be appropriately evaluated and that any potential unreasonable risks can be managed.

In order to carry out its mandates, OPPTS needs the scientific information  to assess and manage
the  potential human health  and ecological  risks  of the various products of biotechnology.
Because of limited resources in the area of biotechnology research, the SP2 program is currently
focused on OPPTS' highest priority of providing the tools and scientific knowledge needed to
understand the  nature  and magnitude of potential  risks and benefits resulting from the use of
genetically engineered  (GE) pesticide products in commerce and the means to prevent or control
any such risks.

The use of biotechnology has led to new pesticide products that control a variety of pests.  These
biologically produced pesticides, which use the inherent pest-fighting abilities of many existing
plants and microbes, have properties that distinguish them from those of conventional chemical
pesticides.  When these products have unique biological properties they may also  pose unique
regulatory challenges.  To address these challenges, the  EPA, USDA, and FDA  have shared
responsibility for regulating agricultural  biotechnology  in the US  (US regulatory agencies
unified  biotechnology  website -  http://usbiotechreg.nbii.gov).   In  particular, EPA regulates
pesticides created through biotechnology as a part of its regulatory jurisdiction over all pesticides
marketed and used in the US. As such, EPA has tailored its basic regulatory framework to fit the
distinctive characteristics of these GE biological pesticides.

In assessing safety, the basic framework for pesticide regulation provides guidance as  to the
nature of any new risks.  Many of the traditional approaches used to assess chemical pesticides
are  applicable to assessing risks from genetically  engineered plants which produce their own
pesticides, also known as  plant-incorporated protectants (PIPs).   PIPs are created when through
the  use of biotechnology, specific genetic material from a bacterium are transferred to a plant to
create plants that produce  pesticidal proteins that the  plant could not previously produce. Under
FIFRA, EPA has the authority to  regulate  the  new protein and its genetic material when it is
pesticidal in nature.  Before making a regulatory decision about a pesticide,  EPA requires data on
a range of subjects to  ensure that the product meets federal safely standards.  For  all pesticide
products, including GE pesticides,  EPA requires testing of product composition and chemical
properties, human health effects, environmental effects  on nontarget pests, and the fate  of the
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pesticides   in   the   environment   (www.epa.gov/pesticides/biopesticides/pips/index.htm).
Therefore,  the research conducted under other SP2 LTGs will be of value  in assessing these
products, to some extent, as well.  However, OPP also recognizes that PIPs may pose uniquely
different risks from traditional, chemical pesticides.    Therefore,  OPP requires  additional
scientific information and tools in order to adequately assess and manage potential risks.  For
example, while there is very low worker exposure and no chemical pesticide spray drift, there are
issues regarding gene flow from PIPs to wild relatives and pollen movement spreading the new
pesticides to non-altered crops. Cross-pollination  of wild relatives can disrupt a local ecosystem
by changing the  makeup of local plants, crowding out related  species and changing the local
habitat. In addition, while the level of protein produced by the newly engineered plant is very
small, because proteins  can be allergens, special emphasis on assessing potential allergenicity is
needed of these products.

From a human health perspective, a major area of uncertainty is on the potential toxicity and
allergenicity associated with biotechnology derived foods.  Potential adverse effects can be from
intended modifications (i.e., from the pesticidal substance)  or from unintended effects resulting
from production of an unexpected substance from  the insertion of the new genetic material into
the host genome.  To date, the  products approved by EPA for use in human food have all been
proteins that degrade rapidly, so no chronic effects would be anticipated. This approach has
been accepted by the FIFRA Scientific Advisory Panel. However, some members of the public
have raised issues about the potential long term exposure to eating foods containing these newly
created proteins.  It is well accepted that the genetic material itself will not cause an acute or
chronic toxic effects and has been exempted from  tolerance.

With  respect to environmental  risk,  OPP needs  effective tools and methods to  evaluate and
subsequently,  if  needed, minimize the likelihood  of negative  ecological effects  such  as  the
following:

Ecosystems
• harm to non-target species, such as soil organisms, non-pest insects, birds, and other animals;
• disruptive effects on specific biotic communities;
• irreparable loss of changes in species diversity and genetic diversity within species.
Agri-Systems
• creating new or more vigorous pests and pathogens;
• exacerbating the effects of existing pests through hybridization with related transgenic plants
   or microorganisms.
Both
• pleiotropic or  epistastic effects on  plant physiology due to emerging metabolic engineering
   approaches.   [These manipulations,  found in  current commercialized transgenic organisms,
   may result in unintended effects in host plants or non-target plants  that may  inadvertently
   receive the transgene.];
• rapid development of resistance to the engineered crop by target pests that may result in greater
   use of more harmful pesticide products over the long term.
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From an ecological perspective, the regulation of biotechnology products is focused  on cases
where there is little prior experience with the new trait and host combination;  a transformed
organism  may persist and perhaps replicate in the environment without  human intervention;
genetic exchange is possible between a transformed organism and unaltered organisms; or the
trait confers  an advantage  to the transformed  organism  over native  species  in  a  given
environment.  Additional concerns about rapid evolution of resistance in targeted pest species;
potential risk to nontarget invertebrates; and alterations in terrestrial or aquatic food chains also
have been raised.

The goal  of the SP2 biotechnology research  program is  to  provide  the scientific information
needed to assess and manage the potential human health and ecological  risks of products of
biotechnology.  The research  will  provide  the  tools needed to generate information  about
biotechnology products and the knowledge needed to understand the nature and magnitude of
potential risks and benefits resulting from the use of biotechnology products in commerce  and
the means to prevent or control such risks.

The biotechnology research  that  ORD is  conducting intramurally or  supporting through  the
extramural STAR program is not only addressing specific  high priority needs identified by OPP
but also is in concordance with needs identified by the National Research Council (2000, 2002).
Furthermore, it is address several science gaps that are noted in the recent  document developed
by the interagency AGRA committee (not publicly released yet).

APG - Provide improved  capability to assess the risks  of allergenicity  of genetically
engineered crops - FY 2011

There  is a concern that as a result of the introduction of novel proteins into the food supply,
biotechnology may unwittingly introduce a potent food allergen that  could seriously affect the
health of susceptible  individuals. OPP is currently unable to adequately evaluate the  potential
allergenicity of proteins introduced into the food supply by gene transfer because valid animal
models to test proteins for potential allergenicity following oral exposure, and other methods to
readily identify proteins that may be potent allergens have not been  adequately  developed.
Without reliable assessment methods, the mechanisms underlying  the development of food
allergy and the factors that contribute to individual susceptibility remain poorly understood.

Research conducted both within ORD's laboratories and through the extramural STAR program
is:
>  Developing an improved understanding of the  basis for human sensitization to dietary allergens and,
    which  in turn, will lead to the development of methods to assess dietary allergenicity, which once
    validated, could become part of the battery of assays that industry must conduct in order to register a
    genetically engineered pesticidal  product.  The projected outcome will be an improved  ability to
    assess the potential risks to human health from genetically engineered foods in the diet and an overall
    improvement in the knowledge of food allergens. 3.1.1
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APG - Provide improved science based risk assessment tools and data support that ensure
improved capability for the comprehensive evaluation of ecological risks and long term safe
use of genetically engineered crops with plant incorporated protectants (PIPs) - FY 2011

The risk of unintended and unexpected adverse impacts on non-target organisms and ecosystems
is  a key issue in environmental risk assessment  of PIP  crop plants.  Research is needed to
examine potential impacts of the effects of Bacillus thuringiensis (Bt) at the field level.  Field
censuses  documenting species  diversity  and  abundance  are  important,  but  they  require
appropriate baseline studies against which  to compare results from agro- and other-ecosystems
containing PIP crop plants. To address these science needs, ORD's research is:

•   Developing standardized and streamlined  methodologies to:  1) conduct base-line assessments of
    agricultural and near-field ecosystems non-target species diversity and abundance to measure direct
    impacts and secondary trophic level effects on non-target organisms, and 2) characterize assessment
    endpoint(s) and the use of predictive strategies to evaluate potential ecosystem level effects.  3.2.1
•   Developing field methodologies to assess and monitor the impacts of the high-dose/structured refugia
    integrated risk management (IRM) strategy on the long-term susceptibility of target  pests to Bt
    endotoxins. The result of this research will be the development of tools capable of identifying the
    evolution of Bt resistance at  sufficiently early stages to allow corrective action to prevent loss of Bt
    crops as effective and least toxic alternatives to conventional pesticides. 3.2.2

APG - Provide guidelines and tools to mitigate gene-transfer and non target effects and the
development  of resistance  in targeted pest  populations  to aid  the  management  of
environmental risks associated with PIP crops to help  maintain the biological integrity of
the environment while minimizing the use of chemical pesticides in agriculture - FY 2015

Laboratory and small scale field testing have been the basis for evaluating the likely safety of
biotechnology  products,  but  long term,  extensive  monitoring has  not  been  conducted  to
determine whether  the effects predicted in such assessments  actually occur in  the field. The
research objectives are to:

       • develop  models to estimate likelihood of insect resistance development that incorporate
       detailed biological information for pest species, including gene flow and mating patterns
       in  the  wild,  geographic  and chromosomal distribution of resistance  alleles, and their
       additive and nonadditive effects on resistance under selective pressures in the field
       • perform  monitoring studies of gene transfer, the development of resistance to PIPs and
       impacts on   non-target  species  to allow  field validation  of  conclusions regarding
       transgenic plants  with new pesticide traits,  including  recommendations to  prevent
       development of insect resistance to support or to refine assessments.
       • develop strategies  for  identifying the  key risks  of  concern  and   effective  risk
       management technologies to  mitigate these  key risks when the monitoring studies
       indicate unintended adverse consequences.

The effectiveness of  strategies for identifying the key  risks  of concern and effective risk
management technologies to  mitigate  these key  risks when  the monitoring studies indicate

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unintended adverse consequences has not been adequately evaluated.  Extensive work is needed
to improve strategies for identifying the key risks of concern and effective risk  management
technologies to mitigate these key risks when the monitoring studies indicate unintended adverse
consequences; and to evaluate whether the genetic alterations produce new  organisms that are
not equivalent to currently existing ones. ORD research is:
•   Developing methods to adequately address the potential for gene flow and introgression to occur and
    the ecological fitness changes  that might result.  Studies are being  conducted to  demonstrate: 1)
    methods for monitoring  pollen dispersal and gene exchange between  crops  and  co-located wild
    relatives and 2) the feasibility of developing effects response information on genes controlling plant
    reproduction, yield, through standard backcrossing and plant community competition experiments.
    3.3.1
•   Gathering population genetic data that will improve the current models used to delay resistance, in
    order to help prevent development of resistance to Bt-corn in Western Corn Rootworm (WCR). ORD
    is using genetic crosses and artificial  selection to identify resistance genes in wild populations of
    WCR across North America.  These approaches will be evaluated for their utility to improve  current
    insect resistance  monitoring  (IRM) plans and for their  utility in risk  assessment of future  PIP
    varieties.  3.3.2
•   Preparing improvements to PIP crop monitoring by:  1) developing methods to identify PIP  corn in
    the field, 2) identifying and assessing the severity of infestation to corn from insect pest populations,
    exploring the use of remotely sensed hyperspectral imagery and less expensive techniques of spatially
    and spectrally  resampling existing hyperspectral imagery  to  simulate  satellite imagery;  and 3)
    applying results from the first two goals and developing an IRM program. 3.3.3
•   Examining the strengths and limitations  of the diagnostic  dose and F2 screen to develop better,
    standardized protocols for use across  the entire Corn  Belt.  Standardized protocols will assist the
    collection of monitoring data for PIP  crops that can be compared  across seasons. The information
    quality improvement undertaken by this research is designed to assist OPP to meet the requirements
    of the Data Quality Act. 3.3.4
•   Determining to what extent simulation models can be used to reliably predict the  onset  of pest
    resistance to PIP  crop controls.  This research has  been designed to provide substantive information
    about the  operation and capabilities of different resistance  management models  to  assist the
    regulatory expert in its proper use and interpretation of results. 3.3.5
•   Starting in 2007, ORD will begin to scope out conducting a cross-laboratory/center effort to develop a
    cost-effective agro-ecosystem monitoring program designed to assess changes in pesticide exposure
    and effect accompanying transgenic crop adoptions. It should be  applicable to a variety of crops,
    transgenic constructs, and spatial/geographical orientations. ORD's initial research goal  will be to
    develop indicators that can be selectively chosen to efficiently  establish causality  relationships
    between transgenic cropping systems, off-farm exposures and ecological responses, thereby providing
    OPP and the industry with an ecological accountability tool. 3.3.6

VII. Relationship to Other Multi-Year Plans

As noted earlier, there are a number of high priority science needs in OPPTS programs which
represent such fundamental and complex scientific challenges that ORD has committed core
research efforts to the problems as follows:
    •  Research in support of pollution prevention issues  can be found in the Sustainability
       Research Program  and  its  MYP,  and therefore,  are  not addressed through  the SP2
       Research Program other than the work that is done related to products of biotechnology.

                                              42

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   •   The scientific gaps in our capability to assess cumulative and aggregate risks, susceptible
       sub-populations amongst human  or  other vulnerable  species,  and stochastic exposure
       scenarios as well as to screen chemicals for their potential to disrupt the endocrine
       systems all comprise additional core  ORD research efforts under the Human Health and
       Endocrine Disrupters MYPs, both of which are providing underlying science and tools to
       OPPTS to meet its mandates, especially those under FQPA.
   •   The need for prioritization tools is  also being addressed by  ORD's core Comp Tox
       Research Program, and to a lesser extent the Human Health Research Program.
   •   Risk assessment frameworks  and methodologies are  developed through the Human
       Health, Human Health Risk Assessment and Ecological Protection MYPs.

Similarly, some of OPPTS'  science needs are being addressed by other problem-driven research
programs as follows:

   •   The scientific gaps pertaining to the toxic effects of respirable dusts are being addressed
       by a comprehensive  Particulate Matter (www.epa.gov/osp/myp/pm.pdf) MYP in support
       of the Office of Air and Radiation.
   •   Most recently, the initiative to understand the implications of nanotechnology on human
       health and the environment, as well as to explore their applications on improving the
       environment are leading to the  development of a new research strategy on the subject.

In addition, the SP2 research program  while specifically developed to support OPPTS needs may
also provide indirect benefits to other Agency Goals and ORD Research Programs.  Examples of
this research include the development of probabilistic spatially-explicit ecological assessment
framework and characterizing the  occurrence of pesticides following drinking water treatment.
Additional examples are included in the table below.
Examples of SP2 Research - Goal 4.4
Prioritization and screening tools
Enhanced interpretation of data
Occurrence of pesticides in drinking water
Releasability of asbestos
Effects, exposure, degradation data on
perfluorinated chemicals
Goals and MYPs that benefit indirectly
Goal 4; Human Health MYP, Comp Tox
Framework
Goals 1-4; Ecosystem Protection, Human
Health, Human Health Risk Assessment,
Endocrine Disrupters MYPs, Comp Tox
Framework
Goal 2; Drinking Water MYP
Goals 1 and 3; Air, Land MYPs
Goals 1-4; Air, Drinking Water, Water
Quality, Land, Human Health, Endocrine
Disrupters MYPs
                                          43

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 Ecological probabilistic risk assessment tools
Goals  2 and  4;  Water  Quality, Ecosystem
Protection MYPs
It should be recognized that, as other related ongoing research has been noted previously, that
there is a need to coordinate the SP2 research with that conducted through other ORD research
programs, in  other federal agencies, and other non-governmental science organizations, and
with our international  counterparts. The mechanisms for collaboration  with  outside-ORD
organizations are highlighted in Section III. In order to improve coordination across the MYPs
within ORD, the NPD for the Pesticides and Toxics Research Program meets periodically with
the NPDs for each of the relevant MYPs as well as the leaders for other programmatic areas
(e.g.,  computational toxicology, nanotechnology, homeland security) who  oversee research
that is ongoing in support of OPPTS.  These discussions are important not only to ensure that
are programs  are not conducting duplicative efforts but also so that we ensure that the products
of the research are disseminated to those who may find them of indirect benefit.

VIII.  Relationship to the Research & Development (R&D) Investment Criteria

  As        part        of       the        President's       Management       Agenda
(http://www.whitehouse.gov/omb/budintegration/pma_index.html),   explicit  criteria  were
developed for managers to use for assessing R&D programs.  The R&D Investment Criteria
consist of three categories, including:

   •   Relevance- R&D programs must  have  clear plans and demonstrate  relevance  to
       national priorities, agency missions, and "customer" needs
   •   Quality-programs should  maximize the quality  of the research through the  use  of
       clearly stated, defensible methods for awarding a majority of their funding
   •   Performance- programs should maintain a set of high priority, multi-year R&D
       objectives with annual  performance outputs and milestones that show how one or more
   outcomes  will be reached

 The  R&D Investment  Criteria will  be  used by two  separate groups to evaluate the SP2
research  program in FY07.   On  February  7-9,  2007 a subcommittee of ORD's Board  of
Scientific Counselors will meet to address a number of charge questions,  including being
asked comment specifically on how the program meets the R&D investment Criteria. OMB
will also use  the Criteria in the evaluation of the SP2 research  program using the Program
Assessment Rating Tool (PART) during FY07.

IX. Communication
    Because  of the breadth of the research program on SP2, effective implementation of this
MYP  requires extensive coordination and communication at multiple levels and multiple
stages. Dedicated coordination and communication is crucial to effective research planning,
allocation of resources, and implementing a cohesive intramural and extramural research
program that is targeted to achieve both the breadth and depth of balance needed to address the
multiple environmental problems that are addressed by the SP2 program.
                                         44

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During planning of the research:
   •   Coordinate identifying the highest priorities for research through a research planning
       committee that includes ORD representatives from the Laboratories/Centers/Offices
       and OPPTS and Regional scientists; ORD, OPPTS, and Regions are partners in
       planning the program.
   •   Communicate priorities to other ORD, OPPTS, and Regional senior managers.

During conduct of research:
   •   Coordinate and communicate across branches and divisions within a particular ORD
       Laboratory/Center where research is addressing a common issue.
   •   Coordinate and communicate across ORD National Laboratories/Centers where
       research is addressing a common issue; this includes working with STAR grantees,
       where appropriate.
   •   Keep the client offices and stakeholders aware of the progress of the research through
       meetings and seminars.  Seminars will be scheduled through the ORD-OPPTS seminar
       series that has been ongoing since 2000.
   •   Hold periodic progress reviews or workshops where the intramural and extramural
       researchers will meet with Agency scientists and managers and other clients and
       stakeholders to share study results and build collaborations.

Upon completion of research:
   •   When researchers complete a body of work, e.g., resulting in a publication, meeting a
       milestone or APM, they are responsible for informing the relevant ORD managers and
       transferring the information in an appropriate format to the appropriate stakeholders.
   •   When an APG is  completed, consideration will be given to preparing a synthesis
       document, where appropriate, that integrates findings of all of the research to
       demonstrate how the multiple studies have contributed to meeting the APG.

To facilitate communication within the Agency and with the public, an SP2 Research Program
website will be developed and maintained.


X. References
National Research Council (NRC). Building a Foundation for Sound Environmental
Decisions. Committee on Research Opportunities and Priorities for EPA. National Academy of
Sciences, Washington, DC, 1997.

National Research Council (NRC). Environmental Effects ofTransgenic Plants: The Scope
and Adequacy of Regulation. National Academy of Sciences, Washington,  DC, 2002.

National Research Council (NRC).  Genetically Modified Pest-Protected Plants: Science and
Regulation. National Academy of Sciences, Washington, DC, 2000.
                                         45

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                                     APPENDIX I
               POTENTIAL ADDITIONAL RESEARCH IF RESOURCES
                             INCREASED 10-20 PERCENT

Should additional resources become available for the SP2 Research Program, the planners will
work collaboratively to identify current high priority scientific uncertainties to address.  In FY06,
OMB introduced a pilot program within the Agency, whereby an additional $4.5 M for research
was recommended to address specific Program Office needs for the Air, Water, Solid Waste, and
Pesticides and Toxics offices. In the area of SP2, teams of managers and scientists from across
ORD's laboratories and centers, OPP,  OPPT, OSCP, and the  lead Region for  pesticides and
toxics held a series of meetings to determine  how these additional resources should be used.
Within a  short period of time,  the  multiple parties reached a consensus  on identifying the
research needed and allocating the resources accordingly. The planners used the previous SP2
MYP as the overall framework to guide their decisions.  In a number of instances, the additional
resources  went to accelerate projects already planned.  In other cases, new research that was
complementary to ongoing efforts was identified. Many of these efforts are described within this
MYP.  Others that are more closely related to the Endocrine Disrupters Research Program will
be described in the updated version of that MYP.   This approach  and  partnership resulted in a
portfolio of research that is already having an impact on Agency decisions even just less than a
year after implementation.   Therefore, the  same approach will be  used should additional
resources become available in the future.

For example, there have been initial discussions between ORD and OPPT on using additional
resources  to address a new emerging biotechnology issue on the assessment of the  safety of
fluorescent proteins to human health and the environment.  A draft background paper has been
developed by OPPT that highlights the scientific questions that need to be addressed and how
research would impact their  decisionmaking  process.   A  brief excerpt  from that document
follows as an example of new research needs that could be addressed.

Problem  Statement:  OPPT has  received submissions over the past  several  years  using
microorganisms marked with fluorescent proteins, both green fluorescent proteins  (gfp) and a red
fluorescent protein,  DsRed originally isolated from the jellyfish Aqueorea victoria and from a
reef coral, Discosoma  sp., respectively.   Given the  potential widespread dissemination  of
fluorescent proteins in OPPT-  and OPP-oriented microbial and plant applications, especially with
the DsRed where there is the potential for it to enter the food supply (an application is in  house
which uses a genetically marked bacterium carried by an insect vector that feeds on numerous
crop species), there is concern regarding unknown effects of fluorescent  proteins to human health
and the environment.  Approach:   OPPT proposes that ORD initially evaluate the potential
human health  effects  of a variety of these fluorescent  proteins if they were to  enter the food
supply in  terms of their stability in the human digestive  system and their potential toxicity,
including  allergenicity.   It would also be desirable to evaluate potential  ecological effects of
these fluorescent proteins,   including toxicity to honey  bees and/or other nontarget  species and
the effects of protein acquisition in photosynthetic microorganisms.
                                           46

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Other areas where OPPTS and ORD  have had discussions regarding conducting research to
address unmet science needs include:  1) detection, analytical, and risk management approaches
for dealing with prions and 2) nanotechnology.   For the latter, there is a separate research
strategy under development by a team of ORD and cross-Agency scientists.  OPPTS will be a
major client of that research program and it is quite conceivable that in the future that some, if
not most,  aspects of that research may  become incorporated into the SP2 research program.
Additional resources in SP2 could then be used to expand the efforts beyond what they currently
are.  Regional Offices  have expressed the continued  need for research regarding clean-up
methods for common household items contaminated from pesticide applications
                                          47

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       APPENDIX II




FLOW DIAGRAMS
          48

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       Figure 3 - Linkage and Timeline for APGs to Meet SP2 Long-Term Goal 1
2009



2010



2011



2012



2013



2014



2015

         2010-Develop the
         scientific underpinning
         related to the effects,
         exposures, and risk
         management of specific
         individual or classes of
         pesticides and toxic
         substances that are of high
         priority to the Agency to
         inform Agency risk
         assessment / management
         decisions
2013 - Evaluate and provide
guidance regarding the
sensitivity and predictive value
of current test methods and
those under development for
the improved identification and
characterization of the potential
of environmental chemicals to
cause human health and
ecological risks
                                                           2013 - Develop the scientific
                                                           underpinnings related to the
                                                           effects, exposures, and risk
                                                           management of perfluorinated
                                                           chemicals to inform Agency
                                                           risk assessment / management
                                                           decisions
2015-Developand
validate virtual
chemical and
alternative methods for
risk-based
prioritization and
screening of chemicals
Long Term Goal 1.  OPPTS and/or other organizations use the results of ORD's research on methods, models, and
data as the scientific foundation for:  1) prioritization of testing requirements, 2) enhanced interpretation of data to
improve their human health and ecological risk assessments, and 3) decisionmaking regarding specific individual  or
classes of pesticides and toxic substances that are of high priority.

                                                    49

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           Figure 4 - Linkage and Timeline for APGs to Meet SP2 Long-Term Goal 2
2009



2010



2011



2012



2013



2014



2015

2009 - Provide approaches
for evaluating the relative
risks from chemical and
nonchemical stressors on
spatially structured wildlife
populations across large
areas or regions, and
provide methods for
characterizing population-
level risks of toxic
chemical to aquatic life and
wildlife








2012 - Provide
improved methods
to assess direct
and indirect risks
to non-target plant
species and plant
communities from
pesticide use

i









2013 -Provide
methods for
extrapolating
toxicological data
across wildlife
species, media,
and individual-
level response
endpoints
i
r
L
1

20 15 -Provide
methods for
characterizing
population-level risks
of toxic chemicals to
aquatic life and
wildlife


A
r i

r
     Long Term Goal 2:  OPPTS and/or other organizations use the results of ORD's research as the scientific foundation
     for probabilistic risk assessments to protect natural populations of birds, fish, other wildlife, and non-target plants.
                                                     50

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      Figure 5 - Linkage and Timeline for APGs to Meet SP2 Long-Term Goal 3
2009



2010



2011



2012



2013



2014



2015

Provide improved
capability to
assess the risks of
allergenicity from
genetically
engineered food

Provide improved
science based risk
assessment tools
and data support
that ensure
improved
capability for the
comprehensive
evaluation of
ecological risks
and long term safe
use of genetically
engineered crops
with plant
incorporated
protectants (PIPs)


T
                                                                                              Provide guidelines and
                                                                                              tools to mitigate gene-
                                                                                              transfer and non target
                                                                                              effects and the
                                                                                              development of
                                                                                              resistance in targeted
                                                                                              pest populations to aid
                                                                                              the management of
                                                                                              environmental risks
                                                                                              associated with PIP
                                                                                              crops to help maintain
                                                                                              the biological integrity
                                                                                              of the environment
                                                                                              while minimizing the
                                                                                              use of chemical
                                                                                              pesticides in
                                                                                              agriculture
Long Term Goal 3:  OPPTS and/or other organizations use the results of ORD's biotechnology research as the
scientific foundation for decisionmaking related to products of biotechnology.
                                                51

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                           APPENDIX III

              Annual Performance  Goals/

            Annual Performance Measures

                          TABLE 1.

Long Term Goal 1. OPPTS and/or other organizations use the results of ORD's research
on methods, models, and data as the scientific foundation for:  1) prioritization of testing
requirements, 2) enhanced interpretation of data to improve their human health and
ecological risk assessments, and 3) decisionmaking regarding specific individual or classes
of pesticides and toxic substances that are of high priority.

APG
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
Annual Performance Goals and Measures
Develop and validate virtual chemical and alternative methods
for risk-based prioritization and screening of chemicals
Report on the development of in vitro models for determining the
effects of chemicals on T4 synthesis
Release the ASTER system via the EPA intranet
Report on approach to evaluate and enhance metabolism simulator
performance through incorporation of metabolic maps for reaction
types and chemicals on EPA priority lists
Report on inventory ranking for OPP lists
Release the ASTER ranking module
Report on cross-species protein expression recognition models in
fish in which are predictive of chemical mode of action affecting
reproductive processes
Provide ORD and the Program Offices with evaluated techniques
for characterizing temporal and compensatory aspects of exposure
using NMR-based metabolomics
Report on using a sensitive Japanese medaka (Oryzias latipes) fish
model for endocrine disrupter screening
Report on a high throughput zebrafish embryo gene expression
system for screening endocrine disrupting chemicals
Report on development and application of a bioluminescent yeast-
reporter systems for screening chemicals for estrogenic and
androgenic effects
Report on Sertoli cell based screening assay
Report on mechanistic approach to screening chemicals and
mixtures for endocrine activity using an invertebrate model
Report on prioritization approaches for multiple inventories and
endpoints
Update ASTERtoxicity pathway identification software with
substructures associated with new pesticide active ingredients
Year
2015
2007
2007
2007
2008
2008
2008
2008
2008
2008
2008
2009
2009
2010
2010
Lab/Center

NHEERL
NHEERL
NHEERL
NHEERL
NHEERL
NHEERL
NERL
NCER
NCER
NCER
NHEERL
NCER
NHEERL
NHEERL
                                52

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APM
APM
APM
APM
APM
APM
APM
APM
APM
APG
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
Report on quantitative, high-throughput assays of
neurodevelopmental processes for cell-based and non-mammalian
models
Report on Biomarker identification, characterization and potential
applicability to high throughput reproductive toxicity testing
Provide ORD and OPPTS with evaluated techniques for screening
large chemical lists using NMR-based metabolomics
Report on the development of a systems model of the thyroid axis
ofXenopus
Identify genomic, proteomic and or other throughput approaches
that improve the efficiency and effectiveness of immunotoxicity
testing
Report on linking proteomic biomarkers to traditional reproductive
endpoints used in chronic fish bioassays
Report on enhancement of metabolic simulations and use in
hypothesis formulation and testing
Provide OPPTS with documented markers of chemical exposure
for selected high-priority classes of pesticides and toxic chemicals
Report on predictive ability of a test battery for developmental
neurotoxicity
Evaluate and provide guidance regarding the sensitivity and
predictive value of current test methods and those under
development for the improved identification and
characterization of the potential of environmental chemicals to
cause human health and ecological risks
Deliver prototype software, input templates, and demonstration
database for capture of pesticide metabolism pathway data.
Report on an evaluation of the use and relevance of the individual
components of DNT, with recommendations for revisions
Report on progress toward building metabolism and degradate
data systems data.
Report on the application of genomics and proteomics for
characterizing the mode of action for conazole-induced mouse
liver tumors and their relevance for human health risk
Report on estrogen elicited gene expression network elucidation in
the rat uterus
Report on systems biology modeling of fathead minnow response
to endocrine disrupters
Report on chemical induced changes in gene expression patterns
along the HPG-axis at different organizational levels using a small
animal model (Japanese medaka)
Report on proteomic approaches using small fish models to predict
chemical mode of action in chemical mixtures
Report on assay fidelity in predicting reproductive toxicity
Report on New Jersey Research Center for Environmental
Bioinformatics and Computational Toxicology
2010
2010
2010
2011
2012
2012
2012
2012
2014
2013
2007
2008
2009
2009
2009
2009
2009
2010
2012
2012
NHEERL
NHEERL
NERL
NHEERL
NHEERL
NHEERL
NHEERL
NERL
NHEERL

NHEERL
NHEERL
NHEERL
NHEERL
NCER
NCER
NCER
NHEERL
NHEERL
NCER
53

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APM
APG
APM
APM
APM
APM
APM
APM
APM
APM-
R
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
Report on Carolina Environmental Bioinformatics Research
Center
Develop the scientific underpinnings related to the effects,
exposures, and risk management of perfluorinated chemicals
to inform Agency risk assessment/management decisions
Develop a database for the PFAA content in new AOC by
combining the data from this project and those published in peer-
reviewed journals
Deliver to OPPT a summary of the concentrations of
perfluorinated compounds (PFCs) in soil samples collected from
around the United States and globally to be used in a risk
assessment regarding PFCs in the environment
Provide data to OPPT on the major PFAA sources in the indoor
environment and the PFAA emission rates from AOC based on
accelerated aging tests and other migration tests
Comparative profiles for developmental toxicity of various PFAA
pertinent to risk assessment
Define the PFAA doses required for different adverse effect
endpoints for determination of BMD
Relative sensitivity of developing liver to PFAA
Reporting on the effects of PFAA on developmental expression
and activation of PPAR-signaling pathways
Deliver to OPPT a series of methods for describing the distribution
of PFCs in key environmental and biological media and for
characterizing potential human exposures to these compounds
Experimental Methods to Evaluate the Stability of Fluorotelomer-
based Polymer Products
Evaluation of fluorochemical degradation during aerobic
wastewater treatment (OECD 303a)
Characterize the immunotoxic potentials of PFAA and the role of
PPAR signaling pathway in the PFAA-induced immunotoxicity
Pharmacokinetic models for selected PFAA for interspecies
extrapolation
Deliver to OPPT a summary evaluating degradation of
fluorotelomer-based polymer products (FBPPs) in soil materials to
be used in a risk assessment regarding FBPPs
Provide data to OPPT on PFAA precursors (e.g., fluorotelomer
alcohols) in the indoor environment and their implications to
PFAA exposure
Comparative pharmacokinetic profiles for selected PFAA
pertinent to risk assessment
Description of hepatotoxicity of selected PFAA and the biological
pathways involved
Determining the effects of PFAA on PPAR-signaling functions
that impact prenatal and postnatal growth regulation
MO As for tissue dysplasia/tumor formation following early life
exposure to PFAA
2012
2013
2007
2008
2008
2009
2009
2009
2009
2009
2009
2009
2010
2010
2010
2010
2011
2011
2011
2011
NCER

NRMRL
NERL
NRMRL
NHEERL
NHEERL
NHEERL
NHEERL
NERL
NRMRL
NRMRL
NHEERL
NHEERL
NERL
NRMRL
NHEERL
NHEERL
NHEERL
NHEERL
54

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APM
APM
APM
APM
APM
APG
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
Extend the evaluation of developmental toxicity of other PFAA
deemed relevant by OPPT
Wastewater screening study to evaluate FC loadings to the
environment
Profiles of hormonal disruption induced by PFAA and their
implication for human health hazards
Evaluation of fluorochemical degradation during anaerobic
wastewater treatment (OECD 311)
Common modes of action for PFAA developmental toxicity
Develop the scientific underpinning related to the effects,
exposures, and risk management of specific individual or
classes of pesticides and toxic substances that are of high
priority to the Agency to inform Agency risk
assessment/management decisions
Provide the Program Office and Regional Asbestos Risk Assessors
with enhanced exposure tools for assessing human exposures to
asbestos in soils and air.
Provide OPPT with a report incorporating the science needs
outlined in the issue paper and workshop discussions on the
development of Pb test kits
Asbestos releasability data and model development completed for
use by Regional Offices and others for greater understanding of
expected asbestos airborne concentrations levels from various
sources under certain environmental and typical
disturbance/activity conditions
Provide OPP with information that will inform guidance to the
public regarding the ability of coating products to reduce
dislodgeable CCA residues on the surfaces of CCA-treated wood
Provide OPP with data that may inform the CCA risk assessment
Evaluated protocols submitted to OPP
Treatment study results submitted to peer journals and to OPP on
acetochlor, molinate and terbufos, on carbamate pesticides, and on
triazole degradates
Provide OPP with data summarizing the enantioselectivity,
exposure, transformation and effects of conazoles and other
modern chiral pesticides for use in risk assessment and regulatory
activities
Provide OPPT with a summary report on the laboratory research
conducted to modify the performance of Pb test kits with the
recommendation(s) for a modified Pb paint test kits(s) that can be
readily commercialized to support the Pb Renovation,
Remodeling, and Painting rule
Provide OPPTS with evaluated tools for forecasting the fate of
pesticides and toxic chemicals in drinking water treatment systems
Treatment study results submitted to peer journals and to OPP on
acetochlor, molinate and terbufos, on carbamate pesticides, and on
triazole degradates
2011
2011
2012
2012
2013
2010
2007
2007
2007
2007
2007
2007
2007
2008
2008
2008
2008
NHEERL
NRMRL
NHEERL
NRMRL
NHEERL
APG
NERL
NERL
NRMRL
NRMRL
NRMRL
NRMRL
NRMRL
NERL
NERL
NERL
NRMRL
55

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APM
APM
Provide OPPTS with a report highlighting the fate of
perfluorinated chemicals under simulated drinking water treatment
conditions
Provide a compendium of AHS Pesticide Exposure Study results
to OPPTS, NCI, and NIEHS for use in assessing and refining
exposure classification in an important agricultural
epidemiological cohort and to provide information about
exposures and related factors.
2009
2009
NERL
NERL
56

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                              TABLE 2.
Long Term Goal 2: OPPTS and/or other organizations use the results of ORD's research
as the scientific foundation for probabilistic risk assessments to protect natural populations
of birds, fish, other wildlife, and non-target plants.

APG
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APG
APM
APM
Annual Performance Goals and Measures
Provide methods for extrapolating toxicological data across
wildlife species, media, and individual-level response
endpoints
Report on quantitative model to estimate in vivo metabolic rates
for fish from microdialysis sampling data
Guidance document for performing histopathology on gonadal
tissues from small fish exposed to endocrine disrupters
Report on protocol for assessing transgenerational effects of
reproductive toxicants on the HPG axis in medaka
Demonstrate a systems-based approach to utilizing toxicogenomic
data to extrapolate across species and biological levels of
organization
Report on and deliver web-ICE and validated ACE to OPPTS to
support pesticide risk assessments.
Evaluate species, endpoint, and associated hazard for ECOTOX
data used in the Pesticide Re-registration process
Demonstration of an approach for predicting population-level
impacts in fish based on molecular responses to chemicals with
different toxic mechanisms of action
Report on the utility of in vitro metabolic assays to predict in vivo
metabolism in fish
Genomic characterization of toxicity pathways in fish as a basis
for extrapolation across species
Report on and deliver updated web-ICE and ACE to OPPTS to
support pesticide risk assessments.
Evaluate species, endpoint, and hazard for ECOTOX data
associated with listed USFWS threatened and endangered species
Systematic, global analysis of biological networks to facilitate
extrapolation across species and chemical exposures in support of
population-level risk assessments for fish
Assessment of the utility of short-term reproductive and
developmental assays with fish for predicting adverse population-
level effects
Provide methods for characterizing population-level risks of
toxic chemicals to aquatic life and wildlife
Methods for assessing the quality of published demographic
parameters for use in population-level risk assessments
Review of published methods and models to incorporate genetics
into population viability models
Year
2013
2007
2007
2007
2007
2008
2008
2008
2009
2009
2010
2010
2011
2012
2015
2007
2007
Lab/Center

NHEERL
NHEERL
NHEERL
NHEERL
NHEERL
NHEERL
NHEERL
NHEERL
NHEERL
NHEERL
NHEERL
NHEERL
NHEERL

NHEERL
NHEERL
                                     57

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APM
APM
APM
APM
APM
APM
APM
APG
APM
APM
APG
APM
APM
APM
APM
Methods for translating existing avian toxicity data into estimates
of change in demographic parameters for use in population models
Provide guidance to program office on the development,
application, and interpretation of simple modeling approaches in a
regulatory context for risk assessment.
Development of stochastic population modeling approaches for
risk assessments of aquatic and avian populations
Report on and provide spatial population models for coastal fish to
OPPTS to support pesticide risk assessments.
Evaluate methods to incorporate complex ecological processes
into population models
Report on and provide improved spatial aquatic population
models, including expanding to additional species, stressors,
locations, and fish communities
Provide guidance on the development, application, and
interpretation of population models to tiered assessment process
Provide approaches for evaluating the relative risks from
chemical and nonchemical stressors on spatially structured
wildlife populations across large areas or regions, and provide
methods for characterizing population-level risks of toxic
chemical to aquatic life and wildlife
Web-accessible, GIS with information on cropping practices,
pesticide use, and avian demographics
PATCH II framework version (Windows) with a generalized life
history module, a general stressor module, and GIS accessibility
Provide improved methods to assess direct and indirect risks
to non-target plant species and plant communities from
pesticide use
Input for protocol for measuring reproductive and developmental
endpoints with annual species
Development of molecular methods for tracking exposure and
assessing effects of plants exposed to low-dose, high-potency
herbicides
Refined regional assessment tools for probabilistic assessments of
risks to plants from herbicides based on GIS framework
Methodologies to determine how chemical stressors and natural
factors control native plant populations
2008
2009
2010
2010
2012
2014
2014
2009
2007
2008
2012
2008
2008
2009
2011
NHEERL
NHEERL
NHEERL
NHEERL
NHEERL
NHEERL
NHEERL

NHEERL
NHEERL

NHEERL
NHEERL
NHEERL
NHEERL
58

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                            TABLES.
Long Term Goal 3: OPPTS and/or other organizations use the results of ORD's
biotechnology research as the scientific foundation for decisionmaking related to products
of biotechnology.

APG
APM
APM
APM
APM
APM
APG
APM
APM
APM
APG
Annual Performance Goals and Measures
Provide improved capability to assess the risks of allergenicity
from genetically engineered food
Develop and use animal models to assess potential risks of
allergenicity associated with plant incorporated pesticides
(PIPs)
Report on improved animal model for assessment of
allergenic potential of foods through selective deletion of T
cells and global gene expression analysis
Report on risk assessment of food allergenicity by a data base
approach
Report on delineation of appropriate specific and targeted IgE
serum testing to assess the potential allergenicity of proteins
introduced by genetic engineering
Report on safety assessment of dietary proteins for
allergenicity using an adjuvant-free mouse model
Provide improved science based risk assessment tools and data
support that ensure improved capability for the comprehensive
evaluation of ecological risks and long term safe use of
genetically engineered crops with plant incorporated
protectants (PIPs)
A scoping meeting regarding the integrated project with
appropriate EPA and outside experts
External Review Draft report outline appropriate tools for
monitoring resistance development to GM crops in the field and
the sue of target pest ecology to refine insect resistance
management strategies to support OPPTS ecology risk assessments
of risk management techniques
External review draft of report on a conceptual framework for
assessing the ecosystem scale impacts of genetically modified
crops to support OPPTS risk assessments
Provide guidelines and tools to mitigate gene-transfer and non
target effects and the development of resistance in targeted
pest populations to aid the management of environmental risks
associated with PIP crops to help maintain the biological
integrity of the environment while minimizing the use of
Year
2013
2010
2012
2012
2012
2012
2011
2007
2007
2008
2015
Lab/Center

NHEERL
NCER
NCER
NCER
NCER

NCEA,
NERL,
NHEERL,
NRMRL
NCEA
NCEA

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APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
APM
chemical pesticides in agriculture
Assessment of pest genetic architecture (population structure and
genetic trait analysis) in order to inform optimized resistance
management plans
Acquire hyperspectral imagery over production fields in Illinois,
Iowa, Nebraska and Minnesota
Generate prototype PIP status and infestation extent maps and
distribute to field personnel for assessment
Detailed field data collection on PIP status, insect infestation levels
and general agronomic conditions of close to a hundred fields
Map assessment for accuracy and refinement based on 2006 results
Acquisition over production fields repeated over a larger area using
more automated map generation methods
SOP for microarray methods for plant samples
Provide report to OPPTS on the development and evaluation of a
gene expression assay of plant incorporated protectant exposure to
non-target insects
Provide report to OPPTS on the genetic characterization of WCR
resistance to CrySBb based on artificial selection and quantitative
molecular genetic approaches
Methods to measure ecological effects of gene flow on plant
communities: herbicide resistant model crops
Provide expert panel recommendations to ORD for designing a
genetic monitoring program to assess long-term effects of PIPs on
non-target organisms
Office of Pesticide Programs (OPP) project involvement
Generate maps over large cross sections of the corn producing
areas of the US and distributed to OPP personnel for their use and
assessment
Molecular indicators of PIP (insect resistance and fungal disease
resistance) gene expression, introgression and population level
markers in GM crop and compatible non-target plants.
Nearly automated production of map development and distribution
for use by OPP, seed registrants and growers for a precise
assessment of PIP status and insect infestations
Incorporate the variation in crop phenology as observed from the
aerial detection including phenological differences between and
within fields.
Methods to measure ecological effects of gene flow on plant
communities: insect resistant model crop
Provide a report to OPPTS on the results of artificial selection for
resistance in WCR- implications for resistance management
programs
Development and assessment of a multi-species molecular assay
for use by Agency scientists to assess exposures to Bt toxins by
target and non-target beetles
Examine landscape configurations involving within field refuge
areas and compare results to whole field evaluations reported

2006
2006
2006
2006
2006
2006
2007
2007
2007
2008
2008
2008
2008
2009
2009
2009
2010
2010
2010
2010

NERL
NRMRL
NRMRL
NRMRL
NRMRL
NRMRL
NHEERL
NERL
NERL
NHEERL
NERL
NRMRL
NRMRL
NHEERL
NRMRL
NRMRL
NHEERL
NERL
NERL
NRMRL
60

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APM
APM
APM
APM
APM
APM
APM
earlier
Each model will be transparently documented for an audience
ranging from professional to public citizenry
Provide OPPTS with an assessment of the results from an ORD
monitoring program to evaluate ecosystem-scale changes
associated with transition to biotechnology -based agriculture
Modeling ecological effects of gene flow from herbicide resistant
and insect resistant GM crops on plant communities.
Methods to measure ecological effects of gene flow from GM
fungal disease resistant crops on plant communities.
Apply the developed technology for resistance monitoring of pests
in cotton production
Methods to measure the ecological effects of GM plants with
stacked PIPs in the presence/absence of selective pressures and
environmental stressors
Refine the methodology that combines analytical and simulation
techniques using initial allele frequencies reported from field
experiments

2010
2012
2012
2013
2013
2014
TBD

NRMRL
TBD
NHEERL
NHEERL
NRMRL
NHEERL
NRMRL
61

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                                     APPENDIX IV
                          DETAILS ON RESEARCH THEMES

                                  Long Term Goal 1:
    OPPTS and/or other organizations use the results of ORD's research on
            methods, models, and data as the scientific foundation for:
                      1) prioritization of testing requirements,
     2) enhanced interpretation of data  to improve their human health and
                           ecological risk assessments, and
  3) decisionmaking regarding specific  individual or classes of pesticides and
                     toxic substances that are of high priority
APG - Develop and validate virtual chemical  and  alternative methods for risk-based
prioritization and screening of chemicals - FY 2015

1.1.1 Screening and Prioritization Models

Research Goals  and Approaches:  Research is: 1) Developing toxicity pathway-based QSARs for
prioritization within large chemical lists, 2) Providing access to peer-reviewed literature and MOA-based
QSAR models  (Assessment Tools for Evaluation of Risk), and 3) Simulating metabolism to enhance
effects modeling.

1) The QSAR pilot  project will use a multifaceted approach to determine the  chemical structural
requirements for initiation of distinct  toxicity  pathways.  It  incorporates  QSAR-based hypothesis
generation, strategic chemical selection for hypothesis testing, in vitro assay optimization and targeted
testing, and QSAR evaluation and improvement for mechanistic  classifications for OPP pesticidal inerts
and antimicrobials, chemicals for which data are lacking and predictions welcomed. The approach is
grounded in seeking  mechanistic understanding of underlying chemical-biological interactions and
defining chemical similarity in terms  of biological activity. The objective is  to present a  process
applicable to recurring issues surrounding determining structural attributes associated with toxicity and
leading to adverse biological consequence. Determinations must be made with enough specificity to result
in reliable predictions but have broad applicability to numerous diverse chemicals. The process used
strives for  mechanistic  interpretability  and transparency, allowing  evaluations of coverage  within
inventories to which models are applied.

2) ORD is collaborating with OPP on the release of ASTER, with its existing battery of MOA-based
QSARs, to EPA's intranet.  Models within ASTER are  upgraded  (e.g., ecotoxicity, environmental
partitioning, environmental persistence, chemical bioconcentration in tissues) and the system capabilities
is expanded  (e.g., ability to search the ECOTOX database for  structural analogs).  This effort puts a
revised version of a tool currently used by the program office directly on their desktops, providing the
program office the capability to rank large lists of chemicals based on  measured data (in ECOTOX) and
predicted values where measured data is absent.
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3) An existing metabolism simulator will be refined to focus on metabolic transformations most likely to
increase toxic potential for an endpoint under study in a related project (Developing toxicity pathway-
based QSARs for prioritization within large  chemical lists). Metabolic transformation types  shown to
enhance estrogenicity, but that currently are underrepresented in an existing simulator, will be studied
using chemicals selected from priority OPP lists. For selected  chemicals, metabolic pathways will be
determined using rat (or fish) in vitro metabolism systems. Analytical methods will be  developed and
used to verify bioactivated metabolites formed in each system under study. Chemicals additionally will be
tested in metabolically competent liver slices  from male fish. Chemical binding to fish ER also will be
verified for chemicals and their  putative bioactivated metabolites  (when available)  in an  associated
project.  Newly generated metabolic maps will be used to retrain the metabolism simulator and,  thus,
increase reliability and  predictivity.  The ultimate goal is to demonstrate an  approach for  predicting
chemical potential for  metabolic  activation.   As  building of metabolism and degradate  databases
progresses (see 1.2.1 - metabolism/degradate databases) they will provide a rich source of metabolic map
information specific  to  OPP chemicals thus  serving as the foundation for developing a metabolism
simulator relevant to reaction types and chemicals of OPP concern.

Impact and Outcomes:  1) The toxicity pathway-based QSAR research will  develop and apply in vitro
and in silico techniques  for prioritization and  ranking within a regulatory context for a defined toxicity
pathway. Model  development and  applicability  is provided with  guidance on use  to enhance data
interpretation. 2) The updating and improvements to the ASTER system will facilitate the identification
of structural analogs and  associated toxicity information to  estimate potential  hazard of untested
chemicals or chemicals  with limited toxic effects information.   3) The metabolism simulator research
helps in better understanding toxicity pathways  from initiating events to  response for metabolically
activated chemicals.  A   computational  tool is provided that  allows prediction and prioritization of
chemicals for which measured data are lacking.

1.1.2 Nuclear Magnetic Resonance (NMR)-BasedMetabolomics

Research Goals and Approaches. ORD will be using NRM-based metabolomics to:  1) understand and
link the exposure and effects  of EDCs within  the HPG axis of small fish models. This includes linking
responses from genomics, proteomics and metabolomics. Information will be integrated in a systems and
population modeling context.; 2) define markers of exposure, and to better understand the cumulative risk
from  triazole  pesticides.  Research  will differentiate responses from triazole pesticides that exhibit
different MO A, assess the impact of metabolism of the parent triazole pesticide on the observed outcome
and determine the extent of conservation of triazole metabolism behavior and metabolomic profiles across
various  species (e.g., fish and rats); 3) identify markers of exposure for important PFCs  such as PFOA
and PFOS  to gain a better understanding of toxic MOA and investigate the occurrence of markers of
exposure from fish collected in contaminated  field sites; 4) develop a HTP metabolomics approach that
involves exposing cell cultures to potentially toxic  chemicals in order to screen large inventories of
chemicals,  such as the HPV chemicals, rapidly for  potential adverse outcomes.  Chemicals found to
greatly alter the metabolite profile of cell cultures (relative to the control case) would be candidates for
more extensive testing; 5) investigate the feasibility of metabolomics to describe the impacts of exposure
of small fish models to nano-materials.  Changes in endogenous  metabolites  would be used  as an
indicator and descriptor of detrimental exposure; and 6) identify endogenous metabolite-based markers of
exposure to toxicants in  human epidemiology  studies in collaboration with the National Cancer Institute
(NCI).  This includes unique sample sets that NCI has collected on occupational benzene exposure in
China and benzidine  exposure in India. (Note: Part of this project is linked with the  second grant under
1.2.4 and 2.1.3)
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Impacts and Outcomes:  The research will result in:   1) validated markers of biologically relevant
chemical exposure to important classes of pesticides and toxic chemicals; 2) information on the temporal
and  compensatory  aspects  of chemicals exposures;  3) information on  similarities and  differences
xenobiotic  metabolism  and the impact  of  exposures  across  key species  (small fish, rats, etc.); 4)
information on the linkage of exposure events to whole organism adverse outcomes. These new methods
of prioritization will be  used by the Program Office to:  1) winnow down the list of chemicals to those
most likely to be of concern to humans and ecosystems; 2) extrapolate impacts  across species where
direct measurements are not available. Furthermore, the markers of exposure that ORD will provide will
be used by Program Offices to determine when effective exposure has occurred and to aid in decisions
about regulating chemicals according to a common MOA.

1.1.3 Screening and Prioritization Methods

Research Goals and Approaches:  ORD research is developing: 1)  Alternative methods for screening
and  prioritization  of developmental  neurotoxicants,  2) cell  culture and biomarker-based screening
methods for nonendocrine reproductive toxicology, and  3) toxicity-pathway-specific protein  expression
models for chemical screening and prioritization

1) ORD will develop in vitro  cell culture models of the  key events in brain development and develop
methods to measure behavioral, morphological and neurochemical outcomes in a limited number of non-
mammalian species. Assays of developmental endpoints will be optimized for quantitative analysis using
HTP technologies. The  predictive capability of the in vitro and non-mammalian  test batteries will be
assessed using a training set of known developmental neurotoxicants, from different chemical classes.
Data will be gathered from the literature and OPP data  sets and will be incorporated into a  searchable
DNT database.

2) ORD will utilize Sertoli cell cultures challenged with a panel of known  reproductive toxicants to
identify insult induced effects in vitro. Sertoli cell markers will be examined and their utility as endpoints
for HTP screening determined. Cell cultures  will also be evaluated using genomic and  proteomic
methodology to identify novel biomarkers suitable for inclusion in the screening model. This approach
subsequently will be extended  to additional reproductive cell lines (i.e., epididymal cells), seminiferous
tubules, and intact animals to  identify additional biomarkers.  Emphasis  will  be on identifying markers
with the potential to be used in  current one generation tier testing.

3) Surface  Enhanced Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (SELDI-TOF MS)
is used to examine protein expression profiling  as a means to screen  chemicals for their MOA. In vitro
exposure of fish hepatocytes  and short-term in vivo minnow exposures  are used to link  diagnostic
expression  profiles  between tissue level  and whole organism assays, and across multiple  fish species.
Culture  media (in vitro) or plasma (in vivo) samples from control and exposed treatments are applied to
Protein  Chip  arrays  to produce protein expression profiles unique  to  each  treatment.  A binary
classification model is constructed from control and treatment protein profiles to identify differentially
expressed proteins predictive of the  MOA of interest. As proof of concept, research efforts are initially
focused on developing diagnostic models with established estrogenic and androgenic chemicals.

Impact and  Outcomes:   1)  A screening approach using cell-based  models and  alternative species
addresses the need to delineate  a strong linkage between responses observed at lower and higher levels of
biological organization.  This research addresses several questions associated with developing a first tier


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approach for screening for developmental toxicity including: 1) are there predictive biomarkers of key
events in the developing nervous system that can be assessed in vitro; 2) how do we apply technological
advances in HTP testing, genomics, and/or  proteomics to develop rapid in vitro screening assays;  3) is
there  a homologous, non-mammalian model of neurodevelopment that can be used as a rapid screen for
developmental neurotoxicity. 2) Screening for reproductive effects using  biomarkers and in vitro cell
cultures will aid in prioritizing the testing of chemicals under  review and characterization  of new
biomarkers will enhance interpretation of existing information on the effects of reproductive toxicants.
This  research will  specifically  approach the needs to utilize  emerging technology to  develop  rapid
screening  and prioritization  tools and to  identify new markers of  effect that would increase our
understanding of current information on reproductive toxicity.  3) Protein expression profiling addresses
the need for targeted in vivo tests based on MOA and targeted omics-based in vitro screens. Profiling the
differential expression of proteins associated with established toxicological pathways, and linking these
profiles across multiple levels of biological organization provides OPPTS with a powerful  predictive tool
for screening  and prioritization of  chemicals. This research  addresses  OPPTS needs  for increased
efficiency and effectiveness of testing programs while providing information necessary to estimate the
toxicological potential of a chemical or chemical class to elicit an adverse outcome

1.1.4 STAR: Development of HTP Screens

Research Goals and Approaches: In 2002 an RFA was released through the Computational Toxicology
STAR program  to  solicit research  proposals that  would lead to the  development of HTP screening
systems for identifying chemicals with estrogen, androgen, or thyroid hormone activities.  The goal was
to develop an extramural portfolio of research on the development of HTP screening systems to assist in
prioritization of chemicals for further screening and testing of their potential as endocrine  disrupters that
would complement ORD's internal efforts. Four STAR awards were made in 2003:

Development and Application of a Bioluminescent Yeast-Reporter System for Screening  Chemicals for
Estrogenic andAndrogenic Effects

Objective:  Researchers have re-engineered the Saccharomyces  cerevisiae YES colorimetric  estrogen
reporter system  to  produce bioluminescence in response to estrogen or  environmental estrogens (S.
cerevisiae  BLYES). Bioluminescence is a reagentless  system  eliminating  the need  for expensive
chromophores.  Light-detection is  more  sensitive than absorbance  detection thus shortening the
development time of the assay.  The colorimetric-based YES has been widely used and is a very useful
tool for assessing estrogenicity  of a  compound or environmental sample. Development of the  BLYES
system has the potential to enhance  its utility. This research is:  1) validating the BLYES system and
developing a standard operating procedure for routine chemical analysis; and 2) developing an androgen
bioluminescent reporter system analogous to the BLYES system.

Mechanistic Approach to Screening Chemicals  and Mixtures for Endocrine  Activity  Using an
Invertebrate Model

Objective: Research is developing a mechanism-based, high-throughput  screening assay  for evaluating
estrogen, androgen, and thyroid (EAT)-like  activities in an invertebrate species that also can be used to
evaluate interactive effects  of endocrine-active compounds through receptor cross-talk.  The  research is
addressing two deficiencies in the Agency's current Tier 1 screening battery for  detecting endocrine
activity of  chemicals:  1)  no invertebrate  screen is  included  despite the  proposed use of a Tier 2
                                               65

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multigenerational test with a crustacean; and 2) the battery is not equipped to assess combined effects of
diverse endocrine toxicants.

A High Throughput Zebrqftsh Embryo Gene Expression System for Screening Endocrine Disrupting
Chemicals
Objective: Researchers are investigating the proposition that perturbations  in the normal  amount or
timing of a hormone-regulated gene product can be taken as evidence of chemical exposure and used as
an end-point in a screening assay to detect potential endocrine disrupting activity using the zebrafish.

Using a Sensitive Japanese Medaka (Oryzias latipes) Fish Model for Endocrine Disruptors Screening

Objective: Research is developing a rapid,  sensitive, biologically-integrated screening assay to identify
EDCs using a sensitive medaka (Oryzias latipes) fish model. Specifically, the research is focusing on two
identified areas  of interest:   identification and  evaluation of EDCs,  and their  categorization  into
(anti)estrogenic, (anti)androgenic, and (anti)thyroidogenic activity.

Impact and  Outcomes:  The research is expected to contribute to the development of HTP screening
systems to assist in  prioritization of chemicals for further  screening and testing of their potential as
endocrine disrupters. The methods generated from these studies will provide  for multiple platforms for
the HTP screening of potential endocrine disrupting chemicals, in  some cases even allowing for the
remote, near real-time monitoring of potential endocrine disrupting chemicals in the environment.

APG - Evaluate and provide  guidance regarding the  sensitivity and predictive value of
current test methods and those under development  for the improved identification  and
characterization of the  potential of environmental chemicals to cause human health  and
ecological risks - FY 2013

1.2.1 Databases for Hypothesis Formulation and Testing

Research Goals  and Approaches:  In  collaboration with OPP, ORD is  developing databases for
structural depiction of metabolic maps and  degradation products.  1) Metabolism. ORD researchers are
developing software  which will allow easy  access to currently  poorly accessible and  unsearchable
metabolic map information that exists  in OPP files. Software  for the rapid  and efficient depiction of
metabolic maps is enhanced to: allow depiction of hierarchical connection sequences of parent chemical
and all listed  metabolites; track radiolabel within a pathway; combine (or separate) maps from associated
radiolabel studies; identify all maps that contain a specific metabolite (or substructure) of toxicological
concern; compare complete maps  across chemicals to find commonalities/differences; compare maps
across species to  detect commonalities, differences, etc. Associated chemical  identifiers,  as well as
bioassay and  analytical chemistry data, and OPP tracking codes are included. Early outputs of the project
demonstrate capabilities of the system by coding metabolic maps for a sample set of pesticides selected
by OPP. Progress is also being made toward population of the database with  OPP data. This is perhaps
the largest collection of metabolic map data collected under the same guidelines, and as such  serves as a
valuable resource to ORD for hypothesis generation and testing to better understand and predict chemical
potential for  metabolic activation (see 1.1.1, part  3 -  Metabolism  Simulator).  2) Degradates. ORD
researchers are optimizing  the metabolic map software to capture chemical  degradate  data. This  will
facilitate the  identification of potentially toxic degradates for the same chemicals assessed for metabolic
activation by OPP, and will enhance the efficiency of these comparisons across the OPP divisions doing
these assessments. As for metabolism data, the system would allow ready access to degradate structures

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associated with parent chemicals, and, where sufficient data permits, to identify pathways of degradation
associated with certain reaction types. Based on name or structure (sub-structure), degradates of particular
concern can be identified; other documented occurrences of the same degradate  (or the substructure of
interest) can be determined; and, the association between degradate and parent chemical structures can be
assessed. This also allows a better understanding of enzymatic reaction types. Initial outputs of the project
will demonstrate capabilities of the system by coding degradate maps for a sample set of guideline study
types selected in partnership with OPP. Because degradates are observed in various study types the
challenge is to determine where sufficient depth of data (i.e., number of chemicals) is available within a
given type of guideline study. This is required to better understanding and eventually predicting reaction
types leading to degradate formation which can be generalized from those which may be assay specific.
Due to these factors, success with metabolism database efforts is prerequisite to determining the detailed
approach and resource  commitment needed to develop a system capable of  predicting degradation
pathways for OPP chemicals (see 1.1.1, part 3 - Metabolism Simulator).

Impact and Outcomes:   1) Access  to metabolism/degradate  data in a searchable format is  key to:
understanding the role of metabolic activation in toxicity pathways; to generating targeted hypotheses that
will address the highest priority uncertainties; and to doing so for the types of chemicals of most concern
to the Program Offices. 2) Electronically stored structures of metabolites/degradates that include pathway
connectivity are necessary first steps to developing and applying genetic algorithms and related artificial
intelligence systems to predict environmental metabolism and degradation pathways. The development of
searchable databases is key to efficient use of existing information and moving toward a new paradigm
based on hypothesis-driven testing and prioritization.

1.2.2 Enhanced Interpretation of Existing Guideline Data

Research Goals and Approaches: Research is being conducted to evaluate and provide guidance  on
major issues regarding the methods used in the current DNT guidelines. Study reports will be reviewed
and summarized in a manner that will allow evaluation of the power and limitations of the data collected
under current guidelines. Parameters  such as variability, reliability, and  sensitivity will  be  evaluated
across studies and laboratories  conducting the tests.  There  are collaborative efforts currently  underway
among multiple organizations in ORD and OPPTS to address these issues.

Impact and Outcomes:  This project will  assist the Agency by enhancing interpretation of data from
DNT studies. The outputs from the project will assist the Agency to revise the DNT guidelines to  provide
better and more accurate data and increase the ability to interpret data. Evaluation of current data also will
identify areas where hazard identification or data interpretation is problematic and, therefore, assist in
prioritization of research to develop new predictive diagnostic markers of DNT.

1.2.3 Identifying Predictive Functional and Molecular Endpoints

Research Goals and Approaches:  ORD is exploring the development of screening assays in the areas
of immunotoxicity and thyroid toxicity and the incorporation of multiple endpoint assessments into a
single integrated testing  protocol.   1)  Research will be conducted to identify sensitive in vivo (using
rodents)  and in vitro approaches  for  recognizing and  screening  for alterations in  immune function,
including immune suppression and chemical allergy. The feasibility of applying genomics as a screen for
alterations of immune function will be pursued. The  potential increased susceptibility of the developing
immune  system, as well as the reproductive and central nervous systems, to environmental chemical
exposure is a major concern  given that these "sensitive" populations would be at greater  risk.   An


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integrated testing approach for developmental neurotoxicity, reproductive toxicity and immunotoxicity
will be  pursued.  2) Research will use in vitro and in vivo exposure systems coupled with genomic
analysis to identify key  events in cancer pathways.  These  new technologies, in combination with
traditional toxicology  experimental systems,  will provide a common accepted set of principles  for
informing extrapolation across test systems, species,  and dose.  These will enable the  design of
targeted and relevant studies to more rapidly predict the  potential for induction of cancer.   3)
Research will use in vitro and in vivo approaches to understand and discriminate the compensatory and
toxicological responses of the highly regulated HPT system. Development of an initial systems model
will be based on the current understanding of the amphibian HPT axis and the compensatory processes
involved in thyroid hormone homeostasis. Experiments  will be conducted to better understand  the
relationships of the critical sub-components of the  system.  Particular emphasis will  be placed on
understanding the relative importance of gene expression  in the pituitary, thyroid, and peripheral tissues
under normal conditions  and  following exposure to chemicals known to interfere with TH  synthesis.
These molecular changes will be linked with functional measurements of key hormones and enzymes that
are part of the HPT pathway, all of which will be interpreted in the context of organismal-level effects.

Impact and Outcomes:   The employment of in vivo and in vitro functional assays, along with  the
application of genomics, should provide a significant link to the identification of potentially undesirable
environmental chemicals that impact the immune, reproductive and central nervous systems, can induce
cancer, and disrupt the endocrine system. A comparison of the sensitivity of in vivo and in vitro function
results versus genomics should lead to the identification of the most sensitive metrics for immuno-, repro-
,  neurotoxicity, or tumor development testing.  Exploration of the conditions under  which  increased
susceptibility of the developing immune system will be an important goal.   In addition, results of this
research will develop a sufficient understanding of the HPT so  that predictive models can be developed,
testing protocols can be abbreviated, and efforts in inter-species extrapolation can be improved.  One of
the most likely uses for  a HPT systems model  is to aid in the understanding and  discrimination of
different MOA. As such,  this work further enables the development of QSARs by providing a basis for
sorting chemicals by mode of action, a necessary step prior to quantifying features of chemical structure
associated with a particular type of toxicity. If these relationships can ultimately be established, then
predictive models can be developed to prioritize/rank chemicals  for future in vivo testing.

1.2.4 STAR: Systems Biology

Research Goals and  Approaches:  In 2003 an RFA was released on Computational Toxicology and
Endocrine Disrupters: Use of Systems Biology in Hazard Identification and Risk Assessment.  The
research goals are: 1) Development of integrative, quantitative models of the function of the HPG or HPT
axes with emphasis on the descriptions  of the  normal physiological processes  and mechanisms  of
perturbation following exposure to xenobiotics  (e.g., EDCs),  in rats  or a  commonly used  small fish
toxicology  model (e.g.,  fathead  minnow, medaka,  zebrafish);   2)  Cross-species   extrapolation  of
integrative, quantitative models of the perturbed HPG or HPT axes following exposure  to xenobiotics
from rats to humans or a commonly used  small fish toxicology model (e.g., fathead minnow, medaka,
zebrafish) to other vertebrates (i.e., within  the same class or across classes). Three STAR awards were
made in 2004:

Chemical  induced Changes  in Gene  Expression Patterns  Along the HPG-axis  at  Different
Organizational Levels Using a Small Animal Model (Japanese medaka)
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Objective: Researchers are developing a screening method to use molecular techniques such as in situ
hybridization, in situ RT-PCR and immuno-histochemical staining to screen for effects of chemicals on
the HPG axis with a special emphasis on steroidogenic pathways and hormonal control mechanisms along
the HPG-axis  in the Japanese medaka. The  methods will allow for  screening of multiple  effects  in
multiple tissues, even at points in development when the tissues are too small to be accurately dissected
for use in more traditional molecular techniques. The methods will be applied to a set of "model" and
"test" compounds for a set of target genes. Once the methods have been developed and validated, they can
be adapted for use with  other genes and/or species of interest and used to efficiently and completely
screen for endocrine disrupter effects beyond simple receptor binding.

Systems Biology Modeling of Fathead Minnow Response to Endocrine Disruptors

Objective: This study is developing  a computational model to evaluate molecular and protein biomarkers
in relation to reproductive  dysfunction in fathead  minnows exposed to environmental estrogens. The
model is incorporating a number of biochemical endpoints along the entire HPG axis, direct evaluation of
physiological changes and reproductive endpoints and the pharmacodynamics and kinetic distribution of
the contaminants. The hypothesis being tested  is that genomic and proteomics biomarkers  will be
diagnostic  of the estrogenic  effects of environmental  estrogens  and  that they will provide a global
understanding  of mechanisms of action that will relate specifically to reproductive endpoints  in fathead
minnow that are adversely affected by exposure to  estrogenic compounds.  This research is being done
through a cooperative agreement with ORD researchers (see research project 2.1.3).

Impact and Outcomes:
This research is expected to result in the development of integrative, quantitative models of the function
of the HPG or  HPT  axes with emphasis on  normal physiological processes  and mechanisms  of
perturbation following exposure to EDCs and improve the ability to extrapolate results across species.

Estrogen Elicited Gene Expression Network Elucidation in the Rat Uterus

Objective: Systems  biology  involves the iterative development  of strategies that integrate disparate
physiological and biochemical data into computational models that are capable of predicting the biology
of a cell or organism. In order to facilitate hazard  identification and risk assessment, a comprehensive
quantitative understanding of the molecular,  cellular, physiological, and toxicological effects that are
elicited  following acute and chronic exposure to synthetic  and natural  chemicals  is required within the
context of the whole organism. This research is developing a computational model that will identify and
predict critical  estrogenic endocrine disrupter elicited changes in gene expression which play a central
role  in the observed physiological/toxic effects based on systematic and quantitative data obtained from
comparative in silico, genomic, molecular and histopathological approaches using a rat uterus model.

1.2.5 STAR: Environmental Bioinformatics Research Centers

Research  Goals and  Approaches:  In 2004  an  RFA was released to  establish an Environmental
Bioinformatics  Research  Center.  Applicants were asked  to focus their proposals in research that is
consistent with the first  and at least  one other strategic objective  of the computational toxicology
program: 1) Improve understanding of the linkages in the continuum between the source of a chemical in
the environment and adverse health and ecological outcomes; 2) Provide predictive models for screening
and testing; and 3) Improve quantitative risk assessment. Two STAR awards were made in 2005. Both
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of these projects  are building bioinformatics  capabilities and  are working  collaboratively  under
cooperative agreements with scientists from ORD and EPA program offices.

The Carolina Environmental Bioinformatics Research Center

Objective: The Carolina Environmental Bioinformatics  Research  Center is bringing together multiple
investigators  and  disciplines,  combining  expertise  in  biostatistics, computational  biology, chem-
informatics and computer science  to  advance the  field of Computational Toxicology. The  Center  is
developing novel analytic and computational methods, creating efficient user-friendly tools to disseminate
the methods to the wider community, and is applying the computational methods to data from molecular
toxicology and other studies.

New Jersey Research  Center for Environmental Bioinformatics and Computational Toxicology

Objective: The Research Center is bringing together a  team of computational scientists, with diverse
backgrounds in bioinformatics, cheminformatics and enviroinformatics, from the University of Medicine
and  Dentistry of New  Jersey, Rutgers,  and Princeton  Universities, and  the FDA's  Center for
Toxicoinformatics. This team is addressing,  in a systematic and integrative manner, multiple elements of
the toxicant Source-to-Outcome sequence (Investigational Area  1,  as  identified in the RFA) as well as
developing cheminformatics tools for toxicant characterization (Investigational Area 2, Predictive Models
for Hazard Identification ). The computational tools will be extensively evaluated and refined through
collaborative applications involving Center scientists as well as colleagues from the three universities and
EPA; particular emphasis will be on methods that enhance current quantitative  risk assessment practices
and reduce uncertainties.

Impact and Outcomes: The research is expected to contribute to development and application of dose-
response information  analysis  and enhance current quantitative risk  assessment practices and reduce
uncertainties.   The computational methods  developed will link to  data from molecular toxicology and
other studies  in order to move risk assessment from a  hypothesis-driven  science toward a predictive
science.

7.2.6 STAR:  Applying Bioinformatics Data  to In Silico Predictive Environmental and Biomedical
Models and Simulations

Research Goals and Approaches:  The goal of the research is to develop predictive environmental and
biomedical computer-based simulations and models that address  data gaps in environmental and human
health risk assessment and will strengthen the ability of predictive scientific data to guide sound future
scientific policy and decisions.  An RFA is being released in Fall 2006 and 3 awards are expected to be
made with research beginning in 2007. The  RFA is being developed in concert with intramural research
efforts and it is expected that some of the awards may be made as cooperative agreements in order to take
advantage of the experience of the intramural investigators.

Through this RFA ORD will seek to fund research  that will synthesize mathematical and computational
models  of biological  systems  that capture  knowledge through the explicit representation  of dynamic
biochemical and biophysical processes  and the application of these models to  understand  complex
biological system functions in response to environmental exposures. Research grant applications driven
by computational and mathematical principle, design, or validation will  be given highest priority.
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Impact and Outcomes: The envisaged long term end product will be the creation of simulations and
models for predicting toxicity pathways and health impacts as a result to environmental exposures and
their use in human health and environmental health risk management.

APG - Develop  the scientific underpinnings  related to the effects,  exposures, and risk
management of perfluorinated chemicals to inform Agency  risk  assessment/management
decisions - FY 2013

1.3.1 Perfluoroalkyl Acids—Developmental Toxicity Characterization

Research  Goals and Approaches:  Because PFOS and PFOA are the most predominant PFCs detected
in humans as well as in the environment, adverse developmental outcomes from in utero and lactational
exposure to these two chemicals will be examined in laboratory rodents.  As PFCs are persistent in the
environment and have long half-lives in humans and laboratory animals, the adverse effects of PFOS and
PFOA will be tracked at various life stages, from perinatal periods to adolescence (puberty), and well into
adulthood.  One of the goals of this research project is to identify an alternative animal model that lacks
the gender difference in chemical clearance that is seen in the rat with some of the PFCs (e.g., PFOA).
The  serum and tissue levels of PFCs will be  determined in the  appropriate animal model  and  these
internal dose metrics will be  used for dose response assessment. Once adverse effects are identified from
the PFOS and PFOA exposure, ORD's investigation will be extended to include other PFCs (such as 8:2
Telomer Alcohol, C-6, C-9, and C-10) that are pertinent to the risk assessment effort at OPPTS.

Impact and Outcomes: This research addresses OPPTS's immediate need to characterize the hazards of
these chemicals for human populations.  Descriptive findings will help  form the basis for complementary
research projects that focus on MOAs for PFAA toxicity.

1.3.2 Perfluoroalkyl Acids—Pharmacokinetic Modeling

Research  Goals and Approaches:  The goals of this research are three-fold: 1) to provide a descriptive
pharmacokinetic profile of PFOA in the mouse (as an alternative model for rat); 2) to discern whether the
pharmacokinetic parameters  of PFOS and PFOA in immature rodents are different from those of adults,
thereby accounting for the unusually high levels of PFOS and PFOA noted in children; and 3) to evaluate
the patterns of PFOS and PFOA  accumulation in various tissue compartments in rodents, and to explore
the  bases  for  bio-persistence of these chemicals in  the body.   This research  is being conducted
collaboratively across multiple ORD organizations. Depending on additional needs from OPPTS, similar
pharmacokinetic studies (particularly those in immature animals) can be extended to other PFAA such as
Telomer alcohols, C-6, C-9, and C-10 compounds.

Impact and Outcomes: These studies will fill an important informational gap to provide a basis for the
margin of exposure (MOE) paradigm of risk assessment of PFOA.  Findings will also provide insights to
the  bio-persistence (long half-lives) of these  chemicals in both animals and humans, as well as the
unusually high levels of these chemicals detected in children.  Studies will be conducted in concert with a
comprehensive investigation coordinated  by NTP, OPPTS and  ORD to examine the  pharmacokinetic
profiles of 18 PFAA identified by OPPT.

1.3.3 Perfluoroalkyl Acids— Characterization  of Immunotoxicity, Hepatotoxicity and Hormonal
Imbalance
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Research Goals and Approaches:  ORD research aims to confirm the reported immunotoxic potential of
PFOA in the mouse, establish NOAEL and LOAEL values, as well as correlate  changes in immune
function  with serum PFOA concentrations.   The investigation will be  extended to include PFOS (and
potentially  other PFCs), and to the rat model.  In addition, developmental immunotoxicity studies will
determine the  relative sensitivity  of the developing and  mature immune systems to  PFAAs and
persistence of effects following developmental or adult exposure.

A few histological features of hepatotoxicity have been described in adult rats after exposure to PFOS and
PFOA.  ORD research  is designed to extend these findings to include biochemical  evaluation of PFCs-
induced hepatotoxicity, including the use of genomic and proteomic techniques to pinpoint  to cellular
insults in the liver, in addition to histological lesions. Hepatotoxic potentials of PFOS and PFOA will be
evaluated in developing rodent models for comparison with the adult, to determine if the adverse effects
are more sensitive during developmental stages,  and if these effects are reversible later in life.

Based on preliminary findings from our laboratory, exposure to PFOS abruptly reduced thyroxine and
triiodothyronine in circulation, without activating the  classical HPT feedback pathway to produce an
elevation of thyroid-stimulating hormone.  ORD's research aims to explore the underlying mechanisms of
these hormonal changes.   More importantly, studies  will  be  conducted to  determine if these
unconventional  thyroid hormone changes are  translated into  physiological dysfunction, and  if these
changes  are relevant to human health hazards. Additional work will investigate if thyroid hormone
imbalance represents a  hallmark response to PFAA  exposure. Furthermore, limited  data have suggested
that PFOA might produce estrogenic effects in the rat. These findings will be confirmed and extended to
elaborate the endocrine  disruptive potentials of PFOA and PFOS (and other PFCs, if warranted).

Impact and Outcomes: This work will identify the hazard potentials of PFCs regarding immunotoxicity,
hepatotoxicity and hormonal interruption and provide OPPT with needed data on additional endpoints for
their risk assessment of these chemicals.

1.3.4Perfluoroalkyl Acids—Characterization ofMOA

Research Goals and Approaches: To evaluate the role of PPAR in PFAA toxicity, ORD will construct
transfected cell  lines with a reporter gene to screen for the specific PPAR activities. PFCs with a high
PPAR affinity will  be  investigated for their ability to alter  lipid homeostasis, fatty acid  transport and
metabolism, placental function, and hematopoiesis.  Gene expression of the PPAR isoforms,  co-factors
and co-activators will be  investigated in various embryonic,  fetal, and placental tissues, and patterns of
changes  in gene expression will be correlated  with cellular endpoints indicating alterations  in growth.
Positive  results  will guide the third phase of our approach, which will utilize a PPAR gene  knock-out
mouse model to confirm  the PPAR-mediated  effects.  In addition, ORD will determine the  specific
involvement of the  PPARa  pathway  in PFOA-induced developmental toxicity, hepatotoxicity and
immunotoxicity by the  use of PPARa-knocked out mouse model.  If warranted, this transgenic mouse
model can be extended to investigation of other PFCs.  MOA studies will also address the role of the
hypothalamic pituitary adrenal (HPA) axis in PFOA-induced immunotoxicity.

Induction of mammary gland tumors,  Leydig cell tumors and ovarian  hyperplasia by PFOA has been
reported  in the rat.  To determine if these tumor  incidences are extended to the mouse model, testis, ovary
and mammary  glands will be  examined in  mice exposed to PFOA during  gestational and  lactational
periods.  Circulating steroid hormone levels and P450 enzyme activity in these tissues will be  monitored
to ascertain whether tumor induction is mediated by disruption of steroid  hormone metabolism.


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Based on the previous observation from ORD laboratories, pulmonary insufficiency is a likely cause for
the neonatal mortality associated with in utero exposure to PFOS or PFOA. In follow-up research, ORD
will attempt to determine the critical window of susceptibility to PFOS-induced neonatal mortality and to
assess the effects of PFOS on prenatal lung development.  Lungs will be collected during the perinatal
period  and  assessed for their degree  of maturity by histological, morphometric,  biochemical, and
molecular techniques. ORD also will measure surfactant proteins in lung lavage fluid from neonates to
assess release of surfactant into the terminal air sacs. Additionally, gene expression analysis of a variety
of markers of lung maturation will be examined for clues into the possible MOA of PFOS.

Impact and Outcomes: Results will provide information regarding the affinity of PFAA compounds for
PPAR isoforms, and comparisons  of the affinities of the compounds in rodent versus human receptors.
This information will be shared with our NTP collaborators for prioritizing the 18 PFCs chemicals in their
investigation.  The PPAR-KO studies will provide  valuable insights into the MOA for developmental
toxicity, neonatal growth and survival, immunotoxicity and hepatotoxicity. The work described here will
also  better define the  target organs (e.g. developing lung)  and the  physiological functions (steroid
hormone metabolism) responsible for the developmental toxicity of PFCs.

7.5.5 Perfluorinated Compounds Methods Development  and  Validation  —  Environmental and
Biological Matrices

Research  Goals and Approaches:  A wide range of matrix specific collection and analytical methods
need to be developed and validated  to begin evaluating environmental distributions, potential human
exposures, and corresponding risks of PFCs.  This research is specifically designed to provide sampling
and analytical methods that will be used to: characterize environmental  distributions of the PFCs; help
determine how humans are  exposed to PFCs; and help evaluate the toxicity, metabolism,  and systemic
disposition of PFCs for the risk assessment process. Emphasis will be placed on developing and applying
methods for water and soil, the two  environmental matrices that  may influence human and ecological
exposures. Priority will also be placed on methods for biological media (e.g., serum, tissue homogenates)
that will measure body burden, disposition, and/or metabolism in ecological and risk assessment studies.
This research will help  provide the methods needed to determine the most important exposure pathways
and risks for the highest priority PFCs.

Impact and Outcomes: This research will meet OPPTS's need to develop  and validate tools (methods
and  protocols)  for characterizing  the environmental  distributions of the PFCs and to assess  which
pathways are important for global transport and human  exposure.

1.3.6 Perfluoroalkyl  Acids—Stability   of  Fluorotelomer-Based Polymers  and  Distribution  of
Perfluorinated Compounds in Soils

Research  Goals and Approaches:  The  stability of the selected FBPPs will be investigated through a
series of laboratory experiments in which FBPPs  are exposed to selected natural and amended soils and
sediment  materials.   Successful  completion  of this  research will include quantitative  analysis of
perfluorinated acids  and alcohols  in complex  soil  matrices at lower detection limits than have been
reported thus far.  Achieving these detection limits entails analyses of these compounds in a wide variety
of soil matrices collected from around the world  to  demonstrate the validity of the methods  being
developed.
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Impact  and Outcomes: The  findings  from this research will  help OPPT develop  a more accurate
assessment of the potential risks posed by PFOA and similar compounds.

1.3.7 Determining the Fate of Fluorotelomer Alcohol-Based Polymer Products During Wastewater
Treatment

Research Goals  and Approaches: The specific goals of the research are to: 1) develop analytical and
experimental methods  to  characterize fluorinated surfactants in various environmental matrices, 2)
describe the composition of FTOH polymer formulations released to the environment via down-the-drain
disposal, 3) determine  the environmental  loadings of FTOHs and PFCs from WW treatment, and 4)
determine the potential for FTOH polymer products to transform or degrade during WW treatment.

Research on other  aspects of FTOH polymer product fate  are planned to address  gaps in the risk
management of these formulations in wastewater. Screening studies of effluents  and  biosolids will be
completed to understand fluorinated surfactant loading to the environment. The fate of these fluorinated
compounds during  anaerobic WW treatment will also  be  explored using simulated anaerobic WW
conditions.   These  studies, along  with  the OPPT  focused work, will  provide a more thorough
understanding of fate of FTOH-based polymers during WW treatment.

Impact and Outcomes: This research addresses OPPTS's request and immediate need to characterize the
environmental stability of FTOH polymer products.  Descriptive findings will advise OPPTS  in their
negotiations with industry [Supplemental Environmental Projects  (SEP)  and  Enforceable Consent
Agreement (ECA)].  The research will  also clarify  if and how FTOH  polymer products contribute to
FTOHs and PFAAs observed in WW effluents and other environmental matrices.

1.3.8PerfluoroalkylAcids —Article Testing and Chamber Studies

Research Goals and Approaches:  The main goals are to characterize the source, transport, and fate of
PFCs in the indoor environment and the  factors that may affect PFCs release from  AOC. This study will
be carried  out in three phases: Phase 1: conduct AOC screening tests to determine how much PFC  is
available in new AOC,  and which AOC are potentially the most important PFC  sources in the indoor
environment; Phase  2: conduct accelerated  aging tests to determine whether AOC can release PFCs to the
indoor environments and, if so, at what relative rates; Phase 3: conduct aging and migration tests under
realistic use and exposure conditions to determine what role AOC may play in human exposure to PFCs,
and the major exposure routes.

Impact and Outcomes: This research was initiated by ORD in response to OPPT's immediate need for
better understanding the sources of PFCs to which the  general population is exposed and  the major
exposure routes in the indoor environment. The findings will close a data gap in  PFCs risk assessment
and suggest potential risk management solutions.

APG - Develop the scientific  underpinning  related  to the effects,  exposures,  and risk
management of specific individual  or classes of pesticides and toxic substances that are of
high priority to the Agency to inform Agency risk assessment/management  decisions - FY
2010

1.4.1 Control of Pesticides in Drinking Water
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Research Goals  and Approaches:   This  research is designed to produce tools  (models, methods,
protocols) to address OPP's drinking water treatment research needs. Specifically,  OPP has requested
that ORD:  1) develop and evaluate a protocol for selected pesticides identified by OPP, and 2) perform
drinking  water treatment  studies for other  selected pesticides  wherein OPP  must  conduct risk
assessments. For both the protocol and ORD's studies, treatment processes will include those commonly
used by drinking water treatment plants and those less commonly used but known or anticipated to be
effective.  These processes include:  coagulation and clarification, softening, adsorption onto powdered
activated carbon (PAC) or granular activated  carbon  (GAC), and  oxidation by chlorine, chloramine,
chlorine dioxide, permanganate and ozone.  Parent pesticides will be studied and, to the extent possible,
transformed pesticides created by hydrolysis  or oxidation will also be  studied.

Originally evaluated using three different types of pesticides, the protocol will also be used as a basis for
providing treatment data for two  classes  of pesticides of interest to OPP - carbamates and fungicide
triazole degradates. Additional classes will be targeted in the future.

ORD also is conducting research  to: 1) provide chemical-specific information on the effects of water
treatment  on   pesticide   transformation   pathways,   2)   provide  physicochemical  parameters  for
transformation products, and  3) develop predictive models for forecasting  treatment effects that cross
chemical class and treatment  conditions. The treatment processes currently being investigated are lime
softening (hydrolysis) and chemical  disinfection (oxidation) as processes with  transformation potential,
and which have been  shown to transform pesticides to either less or more toxic forms.

Impact and Outcomes: This research will:  1) allow OPP to provide an easy-to-use/implement protocol
to pesticide manufacturers for their use in submitting  data to OPP in a consistent manner. It will also
provide data over a  class of  pesticide compounds that challenges the protocol's methods; 2)  provide
drinking water treatment data and its interpretation to OPP for inclusion in their risk assessment, as under
FQPA the contribution of pesticides in drinking water must be considered in the overall risk assessment.;
3) provide OPP with a drinking water degradation model that  can be  used to predict transformation
pathways  across chemical class and treatment conditions;  and  4) provide  OW,  OPP, and OPPT with
treatment data on contaminants that may be  used in their risk assessments. This is a demonstration of a
multi-program opportunity for cooperation in research and implementation.

1.4.2 Reducing Risk Due to Contact with CCA-Treated Wood - Impact of Coatings on Dislodgeable
Chromium, Copper, and Arsenic Residues

Research Goals and Approaches:  The  goals of this work are to  characterize the impact of selected
coating on dislodgeable CCA  residues on the surfaces of CCA-treated wood and, through demonstration
of the approach, provide industry with a methodology that can be used to further develop and demonstrate
the performance of new and improved products. Twelve coating products available  to consumers were
applied to mini-decks constructed from decking materials reclaimed from  in-service decks.  Dislodgeable
CCA residues  were monitored on the surfaces of coated and uncoated  control decks over a two-year
period of outdoor weathering  using a wipe methodology developed  and demonstrated by the  Consumer
Product Safety  Commission (CPSC). In concert with EPA's study, the  staff of the  CPSC conducted  a
similar two-year study by applying eight of the same products to mini-decks constructed from new CCA-
treated wood. EPA and CPSC collaborated on development of the protocol.
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Impact and Outcomes: This research was initiated by ORD in response to OPPT's and CPSC's need for
coatings performance data. The findings will close a data gap in the CCA risk assessment and provide a
demonstrated method that industry can use to further develop and demonstrate products.

1.4.3 Agricultural Health Study - Pesticide Exposure Study
Research Goals and Approaches:   The noted limitations of previous agricultural studies are being
addressed through the AHS, a prospective epidemiological study to quantify the cancer and non-cancer
risks in the agricultural community and to study the relationship between agricultural exposures and
disease.  The study uses questionnaires to provide information regarding pesticide use,  work practices,
and other agricultural exposures, as well as information on other activities that may affect either exposure
or risk for a large  (more than 89,000) cohort of licensed agricultural  pesticide applicators and their
spouses in Iowa and North Carolina.  Information derived from study questionnaires is used to develop
exposure-classification procedures  for subsequent  investigation  of associations  between pesticide
exposure and specific diseases. Exposure measurement data are being developed and analyzed to assess
these questionnaire-based exposure-classification procedures. The AHS Pesticide Exposure Study,  an
exposure-measurement field study  for a subset of agricultural pesticide applicators and participating
family members in the larger AHS cohort, was led by EPA to provide information to assess and refine the
exposure-classification procedures developed from the AHS questionnaire data and to better understand
factors affecting exposures to 2,4-D and chlorpyrifos for agricultural  pesticide applicators and their
families.

Impact and  Outcomes:   The AHS Pesticide Exposure  Study  will improve  exposure  and health risk
assessments made by NCI and NIEHS in the AHS epidemiological study. This will increase the value of
epidemiological study results for OPP use in pesticide exposure and health assessments.  Identifying and
understanding key  exposure  factors can  also  guide  development of improved exposure reduction
strategies and guidance developed by OPP and other organizations. The results may provide information
for pesticide  safety educators on how pesticides can be handled more safely to reduce the exposures to
pesticide handlers and their families.
1.4.4 Asbestos Exposure Research—Air, Soil and Bulk Material Scenarios

Research Goals and Approaches:  ORD is  conducting research to determine which sampling
and analytical approaches best support asbestos analyses.  To address the major gaps in exposure
as identified by the EPA Asbestos Coordination Team, two studies were initiated.  The first
study reviewed the  state of the science to  identify analytical procedures and counting rules
needed for asbestos detection and quantitation in bulk samples, air, settled dust, and soil for field
monitoring and risk analysis in various exposure scenarios. Five areas are specifically addressed
in this investigation: 1)  Identifying analytical  procedures and counting rules needed for asbestos
detection, 2) Comparing  the efficiency of 0.45 jam pore size filters versus 0.8  jam pore size filter media,
3) Rating the efficiency of polycarbonate versus  mixed cellulose ester filters, 4) Recommendations for
bench-level testing  of filters to provide information  on the  performance of these filter media, and5)
Testing on soil/asbestos mixtures. Soils will be  of differing particle-size distributions and organic matter
contents.

The second study addresses the uncertainty in measuring true asbestos fiber concentrations in soil and
bulk materials and estimating possible exposures (e.g. including friability and releasability from these
materials). In particular, research is needed to determine how to separate the asbestos  from the soil while
maintaining the integrity of the asbestos fibers (i.e., without fiber length reduction). An initial method for


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non-destructive separation of asbestos was identified for testing from the peer reviewed literature.  Spiked
soil samples of known asbestos content and asbestos fiber distribution will be prepared and analyzed by
this method. If the method fails to adequately and efficiently separate the asbestos from the soils tested, a
modified method will be developed.

Impact and Outcomes: This research addresses OPPTS's immediate need to characterize the hazards of
asbestos for human populations, especially susceptible populations (e.g. school children) and highly
exposed subpopulations  (e.g. Libby, Montana workers and  families and  users  of nonstochastically
distributed vermiculite products from the Libby mine, as well as areas with elevated concentrations of
asbestos in natural rock formations). The results of the filter data analyses will be used to reduce current
uncertainties resulting from  the use of different filter media and/or filter pore size.  A new method for
sampling asbestos in soils will be developed and provided for use by regional  asbestos risk managers.
The findings will decrease uncertainties in sampling and analysis approaches used in asbestos exposure
and risk assessments.

1.4.5 Asbestos Releasability Research

Research Goals and Approaches:  The goals of this research are  to:  1) develop a framework for
modeling asbestos breathing zone concentrations generated by activities of varying intensity on outdoor
and indoor surfaces and 2) obtain asbestos fiber releasability data from soil and  carpet for calculation of
emission factors.  These data  will  be  obtained from  existing  activity-based sampling (ABS) data,
generation of ABS data, and from instrument tests; i.e., vertical elutriator and/or releasable asbestos field
sampler (RAFS).

Impact and Outcomes:  These data and model(s) will allow EPA Regional  Offices and others to make
rapid decisions about whether a soil or other bulk material is contaminated with asbestos.

1.4.6 Lead Paint Test Kits  Research to Support the EPA "Lead; Renovation, Repair, and Painting
Program; Proposed Rule"

Research Goals and Approaches: To meet the new rule requirements to reduce  exposure to lead hazards
created by renovation, repair, and painting activities that disturb lead-based paint, it is necessary that new
efficient, cost effective technologies meeting the desired sensitivity  within a  specified  range of false
positive and false negatives  rates be developed and made  available  commercially. This research will
initially focus on the investigation of available state-of-the-art analytical technology for its potential to be
modified to meet OPPT's needs for the Lead Renovation and Repair (Pb R&R) rule. The development of
these tests will be guided by the parameters of cost effectiveness, short analysis time,  and field use by
OPPT's primary audience  (remodeler/  building  contractor  complying with  the ruling).   The  kit
configuration should allow  for a prompt commercialization effort. To meet the  short timeframe  (12
months), two concurrent tracks are being  followed: Technical Outreach Support and In-house Laboratory
Research.

An issue paper is being developed describing currently available and emerging Pb test kits, performance
of test kits for Pb in paint (not limited to spot test kits), availability and types  of reference/known  Pb-
based paint materials, and the performance evaluation protocol options. Cost factors of required materials
and protocols are being addressed in the  issue paper. A Workshop is being  planned for Fall 2006. The
workshop objectives include: to obtain additional information on the accuracy, precision and cost of Pb
test kits to determine Pb in paint at the action level; to obtain additional information on the specifications,


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availability, and costs of testing/reference materials to evaluate the performance of test kits for Pb in paint
at the action level; to obtain additional information on the specifications and availability of protocols to
evaluate the performance of test kits for Pb in paint at the action  level; and to determine the cost to
perform these protocols.

Concurrently, in the  laboratory, modifications to adjust the sensitivity and performance requirements of
the currently available test  kits will be investigated. Four areas that impact the  kit's sensitivity and
reproducibility  have  been  identified.  These   areas  can  be  grouped  into   chemistry-based  or
technique/application-based. The four areas that will be experimentally investigated to determine if they
can be optimized or  adjusted, as appropriate, to help the performance of the paint  test kit approach  the
specifications given in the Pb R&R rule are:  1) Paint sampling; 2) Pb extraction from paint; 3) Response
to Pb; 4) Detection of Pb response.  The objectives for each area are:  1) Reproducible paint sampling of a
known area or weight; 2) Quantitative/reproducible extraction for all  Pb-containing paint types; 3) and 4)
Response to Pb and/or detection of the response to Pb adjusted to the action level and reproducible.

Impact and Outcomes: The research findings will be used to help develop a low cost lead paint test kit
that  can be  rapidly commercialized  and  made available to renovators and  remodelers  following
requirements  of the  EPA Pb R&R Rule. It is expected that the developed kit will  be commercially
available within the next three (3) years. The kit, as well as the proposed rule, supports the attainment of
the Federal government's goal of eliminating childhood lead poisoning by 2010.

1.4.7 Chiral Pesticides

Research Goals and  Approaches:  The primary  research  goal  is to determine the environmental
occurrences, fate and effects of enantiomers of selected chiral pesticides, PCBs and other chiral pollutants
with  an emphasis  on currently-used  modern   pesticides  expected to have short  to  intermediate
environmental half-lives.  The approach to  this  research involves  several steps:   1) development of
analytical techniques to separate enantiomers; 2) analysis of environmental and human exposure samples
expected to  contain chiral  pollutants to  determine bioaccumulation and  enantiomeric  ratios;  3)
measurement of enantiomer degradation in selected  environmental matrices to determine selectivity and
rates of enantiomer  degradation; 4)  preparative separation/collection of the enantiomers  of important
pesticides and other pollutants for effects studies; and 5) measurement of the bioaccumulation and effects
of the separate enantiomers using various species and toxicity endpoints, ranging from conventional acute
exposure endpoints  (e.g., LC50),  to  biochemical indicators obtained through application of modern
"omics" tools such as metabolomics and proteomics.

Impact and Outcomes:  This research provides data for OPP to use to better inform risk managers on  the
certainty/uncertainty  and need for additional  information for pesticides that are either single, enriched or
racemic compounds.  The ultimate outcome of this research is to encourage the development, production,
and use of single- or enriched-enantiomer  pesticides  as  a green chemistry activity.   Such pesticide
products would relieve the environment of thousands of tons of unnecessary chemicals that may have
adverse impacts on non-target species, including humans.
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                                   Long Term Goal 2:
  OPPTS and/or other organizations use the results of ORD's research as the
    scientific foundation for probabilistic risk assessments to protect natural
          populations of birds, fish, other wildlife, and non-target plants

APG - Provide methods for extrapolating toxicological data across wildlife species, media,
and individual-level response endpoints - FY 2013

2.7.7 Ecotoxicological Data and Toxicity Estimation Models

Research Goals and Approaches: Research will reduce uncertainties associated with the current use of
ecotoxicity data and existing empirically based interspecies extrapolation models to better estimate the
toxic effects of chemical exposures on wildlife  and aquatic species by:  1) improving accessibility and
usefulness of available toxicity information through the ECOTOX database, a comprehensive Web-based
system maintained by  ORD; and 2) refining empirical models that use available information to predict
toxicity across species (i.e., ICE) and across endpoints (ACE).

Literature acquisition and encoding for the ECOTOX database will  focus on OPP priorities (e.g., studies
on  pesticides  undergoing  reregistration meeting  data quality requirements for  use  in  final  risk
assessments). ECOTOX improvements will include:  1)  providing  chronic effects  data for organic
chemicals for use in the ICE/ACE models, 2) an analysis of toxic effects data available for pesticides that
have undergone re-registration within OPP, 3) an  analysis of toxic effects data for threatened and
endangered species, and 4) a plotting feature for easy comparisons and ranking of hazard across species,
organism life stages, and observed effect endpoints.  Key improvements to the ICE and ACE toxicity
estimation programs will include:  1) identification and expansion of toxicity databases, 2) incorporation
of chemical classes and chemical MOA categories, 3) rigorous model validation, 4)  improved tool
functionality and user guidance, and 5) development of web-based versions of ICE and ACE. Additional
research will evaluate sources of uncertainty in ICE and ACE outputs, including: 1) ACE uncertainty and
protectveness  for chronic mortality  versus other  endpoints,  such as  growth and reproduction; 2)
incorporation of MOA categories into ICE correlation models; and 3) approaches for reducing variability
in toxicity predictions for wildlife and aquatic species.

Existing databases used for extrapolating underrepresented  species, chemicals, endpoints, and lifestages
will be expanded by including additional wildlife and aquatic species toxicity data. Both ICE and  ACE
will be validated by comparing model predictions to measured values. ICE acute toxicity estimates will be
compared to measured LC50 values for multiple species, chemical  classes, and MOA. ACE chronic
mortality estimates will be compared to measured values for no effect and low effects on chronic survival,
growth, and reproduction endpoints determined in available chronic toxicity tests. The results of the
validation and software refinement will be incorporated into new user guidance to allow determination of
the  reliability of both ICE and ACE estimates.

Impact and Outcomes: ECOTOX is used by researchers in the development of high quality models
needed to estimate population effects of toxic chemicals to  wildlife species. The ECOTOX system:  1)
facilitates LTG 2 efforts through quarterly releases of  newly encoded and quality assured data to an
openly accessible database with a robust, user interface for querying and outputting  of data, 2) will be
used to identify data gaps and trends in  existing data sets to assist in furthering research efforts in both
chemical and species extrapolation, 3) will facilitate the identification of structural analogs and associated


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toxicity information to estimate potential hazard of untested chemicals or chemicals with limited toxic
effects information. The ICE/ACE modeling approaches contribute to the ability to predict the effects of
chemicals on aquatic and wildlife  species, an essential component of the conceptual model for ecological
risk assessment. Specifically, this approach facilitates the  estimation of potential  hazard for untested
chemicals or chemicals with limited toxic effects information. Web-based versions of ICE and ACE will
provide immediate action to updates and improvements, and allow greater user flexibility in searching
and correlation analysis.

2.7.2  Mechanistically Based Approaches to Predict Differences in Species Sensitivity

Research Goals  and Approaches: Research will further the development  of  mechanistically based
models to extrapolate toxicity information among chemicals, species and lifestages by:  1) development
and testing of a PBTK model for a fish species,  and an improved experimental method to parameterize
such models; and, 2)  development and testing of an MOA model to predict inter-species differences from
in vitro data.

Three interrelated efforts will advance the development and application of PBTK models for compounds
that undergo metabolic  biotransformation in aquatic species: 1) available information  on in vitro fish
metabolism  will be synthesized and scaled up this for incorporation into PBTK models;  2) experimental
work will be conducted to test the accuracy of model predictions based on in vitro data by making in vivo
metabolism measurements for selected compounds;  and 3) a new experimental system for HTP collection
of metabolism information  will be developed along with  a mathematical model that translates this
information  into in vivo metabolic rate and affinity estimates.

To  examine  potential toxicodynamic differences  among  species,  a pilot project  will be  initiated to
determine the relative differences in insecticide activity at known sites of action in the nervous system.
This will provide a test  of concept that in vitro measurements of effects via a known MOA can predict
species differences in toxicity. Initially, voltage-sensitive sodium channels will be selected as an endpoint,
and the pyrethroids will be utilized as a class of compounds for comparisons. Both the  insecticidal and
acute  toxicological effects of pyrethroids are mediated via voltage-sensitive  sodium channels, and these
channels have been cloned from insects, rodents and humans. These will serve as starting points for cross
species comparisons by obtaining these  clones, expressing them in Xenopus oocytes, and measuring
pyrethroid effects on their function  using electrophysiological techniques.  The ability to predict toxicity
will be determined by comparison  of potency on ion channel function to LD/LC50 values  among the
different species.

Impact and Outcomes: This research will improve the accessibility for higher tiered risk assessment of
mechanistically based methods, which provide  a scientifically-sound basis for extrapolating toxic effects
across species, and identifying sensitive  species. Specifically, the PBTK modeling research addresses
several questions that limit current efforts to extrapolate toxicity data among species, including  1) how
well does in vitro data predict  in vivo rates of metabolism; 2) how variable are metabolism rates within
and among species, 3) under what circumstances does metabolism impact chemical bioaccumulation, and
4) what type of metabolism data has the most utility for incorporation into PBTK models. MOA research
also may provide useful methodologies for predicting  and identifying sensitive populations, and reduce
the need to test all chemicals. In addition, these in vitro methods provide important alternatives to in vivo
and whole animal methods. This research is integrated with ongoing research in the Human Health MYP
as well as related to  cross-species work (animal to  human) that  has been conducted under the air toxics
research plan.


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2.1.3 Extrapolation Methods for Under-Represented Taxa, Lifestages, Chemicals, andEndpoints

Research Goals and Approaches: The overall goal of this research is to utilize three small fish species,
the Japanese medaka, zebrafish and fathead minnow, as a basis for the  development of techniques for
extrapolation of toxicological effects across endpoints, species and chemicals.  To achieve this, a systems-
based approach is being used to define toxicity pathways for model chemicals with well-defined MOA
within the HPG axis.  These pathways serve as a basis for understanding responses of the fish across
biological levels of organization, ranging from molecular alterations to adverse effects in individuals to,
ultimately, changes in population status.  The studies employ a combination of state-of-the-art molecular
biology, bioinformatic and modeling approaches, in conjunction with whole animal testing protocols.

One part of this project involves development of a harmonized medaka multigeneration exposure protocol
that can be used to evaluate population-relevant endpoints (i.e., fecundity, fertility reproductive behavior,
phenotypic and genotypic sex of each generation). This testing also will  include a number of molecular
and histological endpoints diagnostic of MOA and, hence, be useful for  extrapolation across life-stages
and taxa. An important component of this work will be to ascertain the degree to which short-term
(partial life-cycle) assays are predictive of effects in full life-cycle tests.

The other major aspect of this project involves testing with zebrafish, a useful model from the standpoint
genomic analyses, in conjunction with the fathead minnow, the small fish  model most commonly used by
the Agency  for both laboratory testing  and field monitoring. A subset  of chemicals representative of
different HPG MOA will be characterized extensively using a short-term reproduction assay with the
fathead minnow. These results will provide input for population modeling  and provide crucial information
in terms of understanding the consequences of changes in gene and  protein expression and metabolite
profiles with respect to apical responses. Again, data will be collected at  multiple biological levels of
organization thereby supporting quantitative  endpoint extrapolation.  This phase  of the research is being
conducted through collaboration with an interdisciplinary network of EPA (NHEERL,  Duluth; NERL,
Cincinnati, Athens; NCCT, RTP), EPA grantee (University of Florida), and non-EPA partners (e.g., Joint
Genome Institute of the Department of Energy; see second grant description under 1.2.4).

Impact and  Outcomes: This  research will  provide information concerning linkages across biological
levels of organization from molecular to population responses.  Further, the systems-based approach used
to define toxicity pathways in these studies will serve as a basis for extrapolation across species, including
taxa not directly amenable to testing.  Ultimately, the approaches emanating from this work will support
both diagnostic and predictive risk assessments.

APG - Provide methods for characterizing population-level risks of toxic chemicals to aquatic life
and wildlife - FY 2015

2.2.1 Simple Screening Tools to Project Population Responses

Research Goals and Approaches: This effort will explicitly incorporate relatively simple screening
matrix population models into  the  OPP risk assessment process.  Population  models  that have been
developed for OPP will be refined and integrated with the exposure and effects models currently used by
OPP. Some of the factors  for consideration include:  1) the software code, and how to make a seamless
connection between ORD's population models and OPP's existing risk assessment models; 2) translating
OPP effects endpoints into stressor response models that can be used in  the population models; 3) data


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quality for population model parameter estimates; 4) model interpretation, including decisions on which
population model endpoints will are most appropriate for assessing risks; and 5) some level of training
and continuing technical support.

Of particular  emphasis will be the development of a methodology  for evaluating the reliability of
predictions made using population projection models that rely on published demographic parameters.
ORD is using computer simulations that mimic the population-level  risk assessment process  at three
important phases -1) data collection, 2) parameter estimation, and 3) population projection - to attempt to
characterize the risk-assessment scenarios that result in the most serious errors and to provide  a set of
diagnostic criteria that may be used to identify when problems in the  quality of population projections
may be present.

Impact and Outcomes: This  effort begins the important process  of moving from conceptual models of
addressing population response to actual  implementation of population level analyses. Furthermore, this
research advances our understanding of the usefulness of population models for accurately characterizing
risks in ecological risk assessments, and determining if the quality of the population model is sufficient to
answer the risk management question.

2.2.2 More Realistic Projections of Population Responses to Stressors

Research Goals and Approaches: This  effort will provide methods to support probabilistic ecological
risk assessments and will more explicitly  address higher tier ecological risk assessment needs to take into
account greater realism and complexity in projecting population responses to stressors. This will  be  done
using a combination of theoretically and empirically based approaches and field-collected, laboratory-
derived, and simulation-based  information. Selected species will include those used frequently in toxicity
testing,  which historically have provided important information  to regulatory process, including the
registration of pesticides. Research will address four specific areas.
        CD Probabilistic models - Under this research effort, a variety of methods will be explored to
incorporate stochasticity into population modeling.  Data from laboratory and field studies will be used to
explore  useful representations  of variation in population projections. Simulated data also will be used to
test and compare  modeling approaches. (2) Density dependence, and (3) genetics - The implications to
population projections  of the  incorporation  of compensatory  population responses (i.e.,  density
dependence  and changes  in population genetic  structure) also will be explored  using simulation and
experimental approaches.  For example,  a number of data scenarios will be generated using stochastic
simulation models  that are parameterized to  represent the range of scenarios  encountered in  risk
assessment.  Those  density dependence  formulations with the greatest flexibility  and adaptability to
assessment scenarios will be incorporated into EPA's existing suite of population modeling tools along
with guidelines for their selection and use. Also, selected experimental and simulation models will be
adapted and implemented  to  refine and  test hypotheses associated with chemical risks to population
persistence and genetic perturbations. Additional research will use natural populations and a quantitative
genetic  approach  to identify genetic loci important to population persistence during chronic chemical
stress. (4) Spatial effects - Focal species  in ecological risk assessment, as well as their habitats and the
stressors to which they are  exposed, are often spatially heterogeneous and only partially overlapping. As
a proof of the  practical utility  of models for assessing large-scale pesticide exposure and risks in coastal
areas, spatial fish population models will be developed, evaluated and compared using an estuarine fish
species. Published pesticide exposure data will be used initially in the development of demographic
population models, which can  then be expanded into spatial models using literature-based information on
natural history, native habitat, distribution,  range, and occurrence of the  species within  the  Gulf of


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Mexico.  Secondary spatiotemporal  data, such as EPA's  Environmental Monitoring and  Assessment
Program (EMAP)  data, will be incorporated into population-level models with coastal fish species.
Resulting models will be used to project population-level effects in spatial terms and develop an approach
that is intended to have broader applicability to other species, stressors, and coastal areas.

Impact and Outcomes: The application and interpretation of stochastic population modeling approaches
provided through this research supports an important need by OPP for probabilistic, higher tier ecological
risk assessments. This research  will  provide approaches and guidance on the need for and uses of more
complex population modeling approaches as tools for integrating and projecting more realistic effects of
stressors on wildlife populations.

APG - Provide approaches for evaluating the relative risks from chemical and nonchemical
stressors on spatially structured wildlife  populations across  large areas or regions, and
provide methods for  characterizing population-level risks of toxic chemical  to aquatic life
and wildlife - FY 2009

2.3.1 Spatially Explicit Population Models for Avian Wildlife

Research Goals and Approaches: The  research being conducted here will enhance the PATCH model
(Program to Assist in Tracking Critical Habitat) and tie it specifically  to pesticide issues. PATCH is a
spatially explicit, individual-based life history simulator that incorporates GIS representations of real or
hypothetical landscapes. The PATCH model will  be used to simulate  wildlife  population responses to
pesticide application within  agricultural landscapes. Multiple landscape configurations, wildlife  life
histories, and stressor regimes will be explored.

Impact and Outcomes:  The development of a spatially explicit wildlife population model provides a tool
for scaling from individuals up to  populations in  a manner that addresses the complexities of real
landscapes, and that evaluates the cumulative effects of pesticide use and other factors such as habitat
alteration and environmental variability. The model and associated, datasets, documentation, and example
analyses being developed are designed for use by OPP scientists and managers  but are sufficiently
generalizable to be  applicable to the needs of other Program and Regional Offices.

2.3.2 Probabilistic Ecological Exposure Assessment

Research Goals and Approaches:  ORD  is  designing  and implementing a probabilistic exposure
analysis system,  undergirded by a suite of sophisticated process-based models of pesticide  environmental
chemistry and biology, for direct assistance to EPA's regulatory programs in their mandated pesticide risk
assessments.  Research  activities extend from analysis of the  reliability and uncertainty of predictive
modeling tools, through production of user interfaces designed to aid regulatory personnel in their daily
activities, to production of high-quality standardized databases with direct linkages to OPP modeling
software and systematic technological and scientific modernization of terrestrial and aquatic fate and
transport models.  One important objective of this research is to provide OPP  with improved tools for
assessing offsite drift of pesticides, and expanding the capabilities of AgDRIFT and AGDISP to assess
near-field pesticide drift from aerial applications by including source term algorithms  for ground sprayers,
orchard airblast  sprayers, and revolatilization. These new source term models will be embedded in a
transport framework to assess multiple applications  and multiple sources for measuring exposures to a
community or small watershed during a crop production season.
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Impact and Outcomes:   ORD is  developing a scientifically sound approach  to  characterizing  the
exposure element of risk (directly to aquatic resources, and indirectly to humans  through the fisheries)
posed by pesticide contamination of aquatic, estuarine, and marine ecosystems, with credible, state-of-
the-science models that include heretofore neglected areas such as benthic ecosystems, transport across
the benthic boundary layer, and effects of sorption kinetics on ecosystem contamination. Research under
this program generates state-of-the-art improvements in ORD products for use by OPP in implementation
of probabilistic risk assessments.

APG - Provide scientific basis  for  assessments of direct  and indirect risks to non-target
plant species  and plant communities from pesticide use - FY 2012

2.4.1 Effects of Herbicides on Non-Target Plants and Plant Communities

Research Goals and Approaches:  Four inter-related efforts will be conducted.   1) Spatial information
will be  compiled in a GIS  platform which will be made available, at least initially,  to Agency staff
through a Web page, and if successful and feasible, to the general  public.  Research will result in a data
system appropriate for use in risk assessments of effects of pesticides on non-target plants and animals.
Data will be obtained through collaborative efforts with Federal, State, and local agencies or purchased as
necessary. That data will include (as  feasible, but likely not be limited to) political boundaries (states and
counties), human population census data, major geomorphic attributes, crop locations (both conventional
and genetically engineered), resident plants (i.e., noncrop species), bird distributions (from Breeding Bird
Survey data), pesticide use (location  and type of registered pesticides), and wind speed. Additional layers
can be added as requested by OPP or Regional Offices, and, if feasible, outputs will be made available for
use in  wildlife population models.  2) Research will be conducted to provide the scientific basis  for
phytotoxicity (Tier II) testing guidelines with the focus on terrestrial plant effects, with pilot research on
aquatic plants, if possible. Candidate noncrop test species for plant  tests will be based on spatial analysis,
prior use in phytotoxicity testing, ecological significance, and cultural characteristics. These may include
both annual and perennial and native plants. Species will be evaluated to improve traditional seedling
tests and proposed life-cycle tests. 3) Ecological information initially will be obtained  under controlled
greenhouse conditions  to determine the relative herbicide susceptibility of different native species based
on exposure response studies with individual plants growing in pots. Large field-plot studies using these
species  will be conducted to  provide a scientific basis for determining herbicide effects at the plant
community  level. Both  constructed  communities  with planted species  and  in  situ  native  plant
communities would be studied.  4)  ORD will  evaluate whether gene arrays  can  be used  as molecular
indicators as to whether a plant has been affected by specific herbicides, what the possible effects of the
herbicides may be based on the gene activity, and whether this  molecular information can, in the future,
be used to predict the potential susceptibility of different plant species to an herbicide (especially native
plants and threatened and endangered plant species).

Impact and Outcomes: Results of these inter-related efforts will: 1) Improve models  and databases to
improve ecological risk assessments of chemical herbicides to  plants,  plant communities  and possibly
wildlife in a spatially explicit landscape.  2) Produce scientific information for comprehensive and
efficient in vivo assays to evaluate adverse effects of chemical herbicides on individual plant  species,
especially reproductive effects.    3) Produce scientific basis to determine ecological risks of chemical
herbicides on native plant communities, and how they may change over ecologically relevant time periods
as a result of  predicted effects of pesticides.  4)  Produce comprehensive and efficient genomic and
proteomic analysis to identify indicator genes and their metabolic  products (fingerprints) in response to
specific herbicides, and,  as resources  permit,  other pesticides toxic  chemicals  and  stressors. These


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analyses can be used to as a scientific basis for chemical/molecular approaches to characterize risks to
species beyond those initially tested, and for herbicides and other chemicals with similar MOA.
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                                    Long Term Goal 3:
   OPPTS and/or other organizations use the results of ORD's biotechnology
research as the scientific foundation for decisionmaking related to products of
                                       biotechnology

APG  -  Provide improved  capability to assess  the risks  of allergenicity of genetically
engineered crops - FY 2011

3.1.1   Develop Methods to Assess Potential Dietary Allergenicity of Novel Proteins in Genetically
Engineered Food

Research Goals and Approaches:
ORD is addressing the need to develop methods to assess dietary allergenicity and improve understanding
of the  basis for human sensitization to  dietary allergens through a  combination  of intramural and
extramural research projects:

Project 1. Development of an animal model to assess the potential allergenicity of genetically engineered
     food (conducted intramurally)
•    A dietary allergy model in a laboratory rodent is being  developed using a modification of the
    respiratory allergy protocols. Suckling, weanling, and adult rodents (BALB/c or C3H/HeJ mice, or
    Brown Norway rats) are exposed by gavage or injection  multiple times with various  doses of a
    known food  allergen  to  establish the  ability  to  induce  food-allergy  responses.  Allergic
    responsiveness is judged based on the induction of antigen specific IgE, IgGl and possibly IgA, in
    addition to gut mucosal eosinophil influx and respiratory responses.
•   Once the model is developed, rodents will be fed or gavaged multiple times with various doses of a
    prototype transgenic pesticide protein. Allergic responsiveness will be assessed based on results
    obtained from the above studies. Appropriate positive and negative controls will be incorporated into
    the model. The responses to the transgenic pesticide protein allergen in both a purified form and in a
    food matrix will be assessed. The relative potency of transgenic pesticide proteins, when compared
    with known food allergens will be assessed.
•   The model will be:  1)  validated in  several laboratories,  2)  used to assess effects of respiratory
    exposure following oral sensitization and oral exposure following respiratory sensitization, 3) used
    to assess the vulnerability of neonatal and weanling animals relative to adults, 4) used to assess the
    potential influence of toxic contaminants (e.g. aflatoxin)  on the  development of food allergy, 5)
    extended to assess the potential allergenicity of other proteins introduced into the food supply, such
    as fluorescent biomarkers, and 6) used to understand mechanisms and identify potential in  vitro test
    strategies.

Project 2. Solicitations through the STAR extramural program to develop in vitro, in vivo, and in silico
     methods to assess the potential allergenicity of genetically engineered food
The goal of the extramural  STAR research project is to engage the external research  community in
developing methods to assess dietary allergenicity and  improve understanding of the basis for human
sensitization to dietary allergenicity.  One RFA has already been issued calling primarily for specific
research  on safety assessment including:  1)  Development and  evaluation of animal models for safety
assessment;  2) Development of targeted or specific serological assays, and 3) Determination of structure-
activity relationships of allergen proteins.  A secondary area  of interest is research on basic issues
underlying sensitization to food allergens.  The ability to accurately predict the risk of allergenicity posed

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by  the  introduction of a  novel  protein  into the  food supply is currently  impeded  by a  lack  of
understanding of the basic mechanisms underlying the development of food allergy and factors that lead
to susceptibility. These basic issues are not limited to  allergies developing from  GE  food, but have
broader implications for the diagnosis, treatment, and  prevention of,  the  sometimes life threatening
allergic reactions to food. Four grants have been awarded with the first solicitation. Future solicitations
will re-evaluate where the data gaps still exist related to this topic.

Impact and Outcomes:
The research will directly impact the ability of OPPTS  to assess the risk to human health from novel
pesticidal proteins that is required  for regulatory activities related to agricultural  biotechnology. The
research will contribute to EPA's mission by helping  to ensure the safety of the food supply.  The
principal output of this research will be the development of methods to assess potential allergenicity of
novel pesticidal proteins in food. The projected outcome  will be an  improved ability to assess the
potential risks to human health from genetically engineered foods in the diet and an overall improvement
in the knowledge of food allergens. Development and evaluation of animal models suitable for assessing
potential allergenicity  relative to other food proteins  is a high  priority  for  EPA and  other  federal
regulatory agencies. By developing an appropriate animal model, hypotheses regarding conditions (e.g.,
age, genetics) that contribute to susceptibility can be tested. Research on in vitro screening methods will
improve the ability to detect and predict the allergenicity  of both known and novel proteins in the human
diet. Development of a structure-activity database will reduce the reliance on animal  testing and improve
the predictability of allergenicity in humans.

APG - Provide improved science based risk assessment tools  and data support that ensure
improved capability for the comprehensive evaluation of ecological risks and long term safe
use of genetically engineered crops with plant incorporated protectants (PIPs) - FY 2011

3.2.1  Non-Target and Ecosystem Impacts From Genetically Modified Crops Containing PIPs

Research Goals and Approaches:  The goal is to develop methodologies to measure direct impacts and
secondary trophic level effects non-target organisms, and to  characterize assessment endpoint(s) and the
use of predictive strategies to evaluate potential ecosystem level effects.  The risk of unintended and
unexpected adverse impacts on non-target organisms and ecosystems is a key issue in environmental risk
assessment of PIP crop plants. While there has been considerable examination of the effects of Bt crops
on  certain non-target organisms,  particularly using species-specific  laboratory testing,  more work is
needed to examine impacts (or lack of impacts) at the field level.  Field censuses documenting species
diversity and abundance  are important, but they require appropriate  baseline studies against which to
compare results from agro- and other-ecosystems containing PIP crop plants.

ORD  will  develop  standardized  and streamlined methodologies to conduct base-line  assessments  of
agricultural and near-field ecosystems non-target species diversity and abundance.  In addition to broad
field censuses  bio-indicators may be efficient  and sensitive tools to predict  adverse  impacts  during
product evaluation as well as to  measure  the long-term impacts of environmental  releases. ORD will
identify a suite  of ecologically significant indicator species at different trophic levels in, for example, Bt
corn and cotton agro-ecosystems.

Potential impacts of PIP crop plants will  also be examined in terms of ecosystem functions,  such as
nutrient cycling, predator-prey interactions  and the provision of non-target wildlife habitat.  ORD will
develop methodologies and conduct field assessments of these potential ecosystem-level effects in PIP


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crop plants, but expect the results will be relevant to environmental releases of other modified crop plants
as well.

Impact and Outcomes:  The research results will inform regulatory decision-making by OPP and will
provide critical tools to Regional Offices involved in field test and post commercialization monitoring.
The experimental data and critical evaluations will reduce uncertainty and strengthen risk assessments of
PIP crops submitted to the EPA for registration under FIFRA.  Data from this work has been used to
support the drafting of proposed regulations for the Agency.

3.2.2 Field Assessment oflRMforPIPs

Research Goals and Approaches: The goal is to  develop field methodologies to assess and monitor the
impacts of the high-dose/structured refugia IRM strategy on the long-term susceptibility of target pests to
Bt endotoxins. The development of target pest resistance to the Bt transgene[s] used as PIPs is a serious
risk both to the sustainability of Bt crops and to the  wider utility of environmentally 'soft' microbial Bt
pesticides.  Therefore, the EPA requires growers of Bt crops to follow the high-dose/structured refugia
strategy to delay or prevent resistance development.  Effective management requires sensitive tools for
detecting resistance in field pest populations while the resistant alleles are still sufficiently rare to allow
for corrective action.

The research will focus on field testing and  validation of the high-dose/structured refugia strategy for Bt
resistance management.  Key assumptions of the models upon which this strategy is based still have not
been tested in field populations of the target pests.  Significant  data gaps exist regarding  pest biology,
ecology and population dynamics, particularly with respect to dispersal and use of alternate hosts. Target
pests include  key lepidopteron cotton and corn pests, and beetle pests.  ORD proposes to  address these
ecological data gaps in a series of field and regional-scale studies.

Impact and Outcomes:  The research results will inform regulatory decision-making by OPP and will
provide critical tools to Regional Offices involved in  promoting grower compliance with insect resistance
monitoring (IRM) requirements.  EPA will develop tools capable of identifying the evolution  of Bt
resistance at sufficiently early stages to allow corrective action to prevent loss of Bt crops as effective and
least toxic alternatives to conventional pesticides.   Results  from  spatial/temporal  analyses of insect
sensitivities to Bt toxin will be used by OPP  staff in evaluating results from current monitoring efforts and
future management planning.

APG - Provide guidelines and tools to  mitigate gene-transfer and non target effects and the
development of  resistance  in targeted  pest  populations to aid the   management  of
environmental risks associated with PIP  crops to help maintain the biological integrity of
the environment while minimizing the  use of chemical pesticides in agriculture - FY  2015

3.3.1 Ecological Effects of Gene Flow

Research Goals and Approaches: Although  industry has not  yet requested registration  of a PIP in a
plant that has relatives in the US with which it can hybridize, such requests are certain to be made in the
very near future. As with any risk assessment, methods are needed that adequately address the potential
for gene flow  and introgression to occur (e.g.,  pollen dispersal distances; sexual compatibility)  and the
ecological fitness changes that might result (e.g.,  increased weediness;  changes  in reproductive vigor).
Studies were  conducted that demonstrated methods  for monitoring  pollen dispersal and gene exchange

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between crops and co-located wild relatives. Other studies underway in the greenhouse and mesocosms
are demonstrating the feasibility of developing effects response information on: 1) genes controlling plant
reproduction; 2) yield, through standard backcrossing experiments; 3) plant community competition.
Because of significant differences in plant types, two model crops are used: creeping bentgrass, which is a
monocot, perennial, wind-pollinated,  cool season turfgrass and canola, an annual dicot that is primarily
pollinated by insects. ORD used commercially available GE genotypes of each of these crops engineered
to be resistant  to the herbicide glyphosate (Roundup®)  through expression  of the selectable marker
CP4EPSPS. More recently, via academic collaborators, ORD has obtained GE canola resistant to insects
and creeping bentgrass that has disease tolerance.

Impact and Outcomes:  Research from this project has provided the Agency with useful tools for
generally  assessing nontarget  risks   from  PIPs  and has helped inform  Agency decisions  on  the
environmental safety of the products  of agricultural biotechnology, particularly with regard to potential
long term effects on plant communities. Studies on pollen dispersal and gene flow have changed the
paradigm of understanding the distances and probabilities of gene transmission. Protocols for conducting
plant microarrays will allow the Agency to critically review registrant submissions. Lessons learned have
been applied to discussions of rule making for exemption of viral coat protein PIPs. The project  also has
contributed  to  the  general  scientific knowledge of potential  ecological  consequences  of  genetic
engineering through publication  of  journal  articles  and presentations at national and international
conferences.

3.3.2  Long- Term Responses of Targeted Pests and Non- Target Organisms to PIPs

Research Goals and Approaches:  In order to help  prevent development of resistance to B^-corn in
WCR,  ORD will be gathering population genetics  data that will improve current models used to delay
resistance.  In addition, ORD is using genetic crosses and artificial selection to identify resistance genes in
wild populations of WCR across North America.  Molecular genetic approaches are  used to  identify
candidate resistance genes that can then be further characterized  at molecular and physiological levels.
Resistance  genes are used  to  develop  PCR-based assays to monitor populations  based on allele
frequencies  at these candidate genes. These approaches will be  evaluated for their utility to improve
current IRM plans and for their utility in risk assessment of future PIP varieties.

The long-term ecological outcomes associated with Bt-com adoption remains  a concern to EPA.  To aid
in the assessment of long-term ecological risk, a second research goal is to evaluate an approach to  long
term monitoring for potential consequences  of GE crops for non-target organisms.  Specifically,  ORD is
investigating the use of genetic data to 1) develop a molecular assay for detection of biologically  relevant
exposure to CrySBb protein in non-target organisms, based on change in gene expression; 2) estimate
changes in  numbers of effective  breeders and gene flow in non-target populations; 3) provide  a
quantitative and objective measure of biodiversity. The initial approach is to focus on ground beetles and
rove beetles and then beginning in FY09, to expand to other valued ecological endpoints, particularly
beetle predators and competitors, in order to  better assess broader ecological consequences.

Impact and Outcomes: ORD's research has direct relevance to OPP's policy and regulatory decision-
making and addresses public uncertainty about ecological risks of GE crops. Resistance biology research
provides a sound scientific basis to defend and improve  EPA-mandated resistance management plans.
Molecular methods ORD developed for genetic analysis of WCR are already being used by academia and
USDA to  monitor  WCR populations and evaluate  existing population  models. By improving  the
biological realism of models used  to  decide how to delay/prevent insect resistance to Bt-com, ORD's


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work will improve the longevity of Bt -corn products and, hence, their environmental benefits.  Methods
to evaluate long-term, ecosystem-scale outcomes of Bt-corn will allow OPP to make informed decisions
about the eventual consequences of greater GE crop adoption. ORD's non-target monitoring protocols are
expected to be a key component of future ecological assessments designed to evaluate the long-term
ecological costs and benefits associated with GE agriculture adoption.

3.3.3 Improvements to PIP Crop Monitoring through the use of Remote Sensing

Research Goals and Approaches:  The project has three primary goals: 1) develop methods to identify
PIP corn in the field; 2) identify and assess the severity of infestation to corn from insect pest populations.
The  project partners are exploring use of remotely sensed hyperspectral imagery to determine if these
goals can be met, but will also assess whether less expensive, more accessible satellite imagery can be
used by spatially and spectrally resampling existing hyperspectral imagery to simulate satellite imagery;
and 3) take results from the first  two goals  and develop an IRM.  The IRM program will use coarser
resolution satellite imagery to evaluate regional corn growing areas for insect pest infestation potential
and then use the greater resolution provided by hyperspectral imagery to investigate whether resistance is
developing in  specific PIP corn locations. If it appears probable infestations are  occurring in PIP corn,
field crews will be deployed to these locations to sample for PIP resistant insect pests.

Impact and Outcomes: Completion of this research will lead to a number of beneficial outcomes for the
EPA, seed companies and growers, including the development of an improved management plan  for Bt
stewardship that  minimizes  resistance   development  and  maximizes  effectiveness of Bt  toxins.
Additionally, growers will receive feedback on effectiveness  of Bt in their fields and information  on
insect pest damage to their crop if it develops.

3.3.4 Standardization of PIP Crop Resistance Assay Procedures

Research Goals and Approaches  The goal of resistance monitoring is to aid researchers in evaluating
resistance management strategies by determining baseline susceptibility levels to transgenic insecticides,
detecting changes in the resistance allele  frequencies, documenting changes in the level of resistance in
the field,  and  documenting control failures  due to resistant  insects.  Several  methods for monitoring
resistance to Bt proteins have  been proposed for the European corn borer.  Two of these methods have
produced  the  primary data  EPA has  been  using for regulatory  decisions regarding  Bt corn  by
discriminating concentration assays and the F2 screen.  Researchers and regulatory agencies must have
sound  confidence in interpretation of resistance monitoring  results  and understand limitations  of the
various methodologies to make economically  and environmentally sound decisions.  To effectively
monitor frequency of resistance alleles in wild populations of insects, researchers must balance concerns
of statistical precision at  low allelic frequencies,  costs of sampling,  and the organization and labor
required to intensively sample  many individuals.  The purpose of the research is to examine the strengths
and limitations of the diagnostic  dose and F2  screen to develop better, standardized protocols for use
across the entire Corn Belt.

Impact and Outcomes:  ORD is working with USDA to extend the standardization  effort to  cotton
insects. Recent field results have underscored the importance of standardized analytical methodology and
care of field collected insects. Standardized protocols will assist the collection of monitoring data for PIP
crops that can be compared across seasons. The information quality improvement  undertaken by this
research is designed to assist the Office  of pesticide programs to meet the requirements of the Data
Quality Act.


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3.3.5 Evaluation and Testing Simulation Model for the Evaluation of Resistance Development in Pest
Populations

Research Goals  and Approaches:   Methodology has been  developed for combining analytical and
simulation modeling techniques to take advantage of the  best features  of both methods of evaluating
resistance management plans.  By combining the two techniques, the simulation models can be used to
generate input parameters for the simpler analytical models.  Using this  methodology, a wider range of
resistance management  strategies can be evaluated at realistic allele  frequencies.  Preliminary results
suggest that there are several situations where resistance to transgenic crops may not evolve in the pest
population.

Impact and Outcomes:  Verification that an IRM model conforms  to evolutionary theory using the
procedures developed for this project should become a standard for future IRM models.  Sampling from
complex simulation  models to  generate the input parameters  for use in the  analytical models should
significantly reduce the  time required to evaluate resistance management plans.  This methodology will
identify the range of landscape conditions where evolution of resistance is unlikely and provide guidelines
for the landscape conditions that pose the greatest risk of resistance evolution.

3.3.6 Systematic Development of an Agro-Ecosystem Monitoring Program to Assess Environmental
Risks and Benefits ofPesticidal and Herbicide-Tolerant Transgenic Crops

Note: This project represents a cross laboratory cooperative effort that extends from the work done at the
individual labs and center since FY2003. Because this is a transitional project,  a detailed research plan
will be developed in FY2007and actual implementation of the research begun in FY2008.

Annual Performance Goal:  To provide science based risk assessment tools and data support that ensure
the comprehensive  evaluation  and  long term  safe  use of  genetically modified  crops  with plant
incorporated protectants (PIPs).

Problem Statement: Despite widespread adoption of transgenic crops and clear indications of economic
and environmental benefits from recent research, there are sufficient contrary results which, along with
continuing public concern and  the promise of new crop  varieties, warrant new approaches to EPA's
research.  A systematic approach to monitoring PIP crop usage, conventional pesticide exposure, and
associated ecological effects directly has been done only on a limited basis and without a  view to
development of broadly applicable schemes.  Existing  data are not sufficient to clearly establish the
degree to which transgenic crop adoption, separate from other environmental and agricultural trends, has
resulted in improved human health and decreased detrimental effects on non-target organisms at national
or regional scales. The  development of a cost-effective monitoring  program that clearly links transgenic
crop usage to conventional pesticide exposure and ecological effects  will fill this data gap.

Research Theme: This project is a  cross-laboratory effort to  develop a cost-effective agro-ecosystem
monitoring program designed to assess changes in pesticide exposure and  effect accompanying transgenic
crop  adoptions.   It should be  applicable  to a variety of crops,  transgenic constructs,  and  spatial/
geographical orientations. ORD's initial research goal will be to develop indicators that can be selectively
chosen  to efficiently establish  causality relationships between transgenic cropping  systems, off-farm
exposures and ecological responses.  The 7-year objective will be to provide OPP and the industry with an
ecological accountability tool. The monitoring program will be designed to address such questions as:


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    o  To what degree does transgenic agriculture contribute to reductions in broad spectrum chemical
       pesticide applications within the Corn Belt?
    o  Is GE agriculture contributing to an overall reduction in risk to native biodiversity via reduced
       pesticide  exposure  or altered agricultural practices? Does  the proportionate risk to different
       species change with shifts in these agricultural practices?
    o  Does GE agriculture adoption result in a  decrease in human pesticide exposure for agricultural
       communities in the Corn Belt?
The research challenge for ORD  is to develop a rigorous monitoring program that provides  statistical
power at a manageable cost. Development of the program will require input and analysis from a wide
range of disciplines.   Therefore, building upon  the resident  ORD experience,  the  first year will be
dedicated to  development and  organization of a  detailed plan by OPP and ORD personnel and then
followed by outside  peer review.  Although the ultimate design is  contingent on the outcomes of this
detailed  planning, ORD believes the basic framework for  ecological  and  human monitoring  will
incorporate longitudinal studies at fixed monitoring sites to aid  in causality analysis, combined with
probabilistic  sampling of additional sites  for purposes of spatial  extrapolation. Ideally, the ecological
assessment  will  demonstrate  whether expected changes  in weed, lepidopteron  and  coleopteran
communities propagate  into  effects  on  sensitive  vertebrate communities, particularly birds  and
amphibians.

Because  the  design goal is  to  provide trends information on exposures and  effects at minimal cost,
successful development and  application of powerful new indicators  and approaches (e.g., hyperspectral
imaging  analysis,  genomic technologies,  and  molecular population genetic analyses  that estimate
multigenerational responses  and model long term population outcomes at regional scales) are needed.
Partnering with other Federal agencies will also be pursued. A scoping meeting with appropriate EPA and
outside experts will be scheduled in FY2007.
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    APPENDIX V
Key ORD Investigators
Research
Theme
1.1.1
1.1.2
1.1.3
1.1.4
1.2.1
1.2.2
1.2.3
1.2.4
1.2.5
1.2.6
1.3.1
1.3.2
1.3.3
1.3.4
1.3.5
1.3.6
1.3.7
1.3.8
1.4.1
1.4.2
1.4.3
1.4.4
1.4.5
1.4.6
1.4.7
2.1.1
2.1.2
2.1.3
2.2.1
2.2.2
2.3.1
2.3.2
2.4.1
3.1.1
3.2.1
Key ORD Investigators
P. Schmieder, C. Russom
T. Collette, D. Ekman, J. Kenneke, C. Mazur, Q. Teng, T. Whitehead
W. Mundy, J. Welch, M. Hemmer
D. Mustra, NCER Project Officer; Grantees: G. Sayler, G. LeBlanc, G. Callard,
S.Teh
P. Schmieder
K. Crofton
R. Smialowicz, R. Luebke, M Sel grade, S. Degitz
D. Mustra, Project Officer; Grantees: J. Giesy, N. Denslow, T. Zacharewski
D. Mustra, Project Officer; Grantees: F. Wright, W. Welsch
D. Mustra, Project Officer
C. Lau, J. Rogers, B. Abbott, S. Fenton, G. Klinefelter, A. Lindstrom, M. Strynar
C. Lau, A.Lindstrom. M. Strynar, H. Barton
R. Luebke, D. Wolf, C. Lau, M. Rosen, M. Gilbert, C. Gordon
J. Rogers, B. Abbott, S. Fenton, G. Klinefelter, M. Rosen, C. Lau
A. Lindstrom, M. Strynar, C. Lau , J. Washington, M. Mills, ORISE Fellows:
Shoji Nakayama, XiBiao Ye
J. Washington, J. Ellington
B. Boulanger, C. Acheson, T. Holdsworth, M. Mills
X. Guo, M. Mason, K. Krebs, X. Liu
R. Miltner, S. Duirk
M. Mason
L. Sheldon, C. Croghan, P. Jones
D. Vallero, B. Schumacher
G. M. Shaul
S. Harper, J. Van Emon, K. Rogers
W. Garrison, E. Ulrich, M. Tapper
C. Russom, M. Barren, S. Raimondo
J. Nichols, T. Shafer
G. Ankley, R. Johnson, D. Bencic, J. Lazorchak, T. Collette, R. Conolly, Grantee:
N. Denslow
J. Grear, G. Thursby, T. Gleason, R. Bennett, M. Etterson, S. Raimondo
J. Grear, D. Nacci, S. Raimondo, M. Barren
N. Schumaker, R. Bennett, M. Etterson
L. A. Burns, S. A. Bird
D. Olszyk, T. Pfleeger, E.H. Lee
M. Sel grade, C. Bowman, S. Laessig
B. Frederick
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3.2.2
3.3.1
3.3.2
3.3.3
3.3.4
3.3.5
3.3.6
B. Frederick
L. S. Watrud , J. R. Reichman, E. H. Lee, C. Burdick, T. Shiroyama, B. M.
R. Waschmann
Smith,
U. Stolz, S. Franson, B.Daniel, M. Bagley
J. Glaser, M. Carroll
J. Glaser
J. Glaser
B. Frederick, M. Bagley, L. Watrud, A. Fairbrother, J. Glaser
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                                    APPENDIX VI
                                ACCOMPLISHMENTS
Long Term Goal 1
Safe Communities - Research program in LTG 1 prior to 2003.
•   Sensitivity of the Young to Pesticides: ORD research developed data that: 1) identified
    pesticides to which the young are uniquely sensitive; and 2) directly influenced regulatory
    actions and risk assessment decisions for these pesticides (NHEERL).
       o  OPP cancelled or reduced household and agricultural uses of selected cholinesterase-
          inhibiting pesticides to decrease potential for exposure in the young.
       o  OPP issued a Data Call-In (DCI) for all registered organophosphates (-30) to collect
          data on comparative sensitivity of the young.  ORD data were instrumental in
          developing the testing paradigm required by OPP of pesticide registrants for this
          evaluation. This DCI has provided information used by the Agency to evaluate the
          risk to infants and children
       o  ORD research contributed to the development of guidance for selecting appropriate
          age groups for assessing childhood exposures to pesticides.
•   Mixtures: ORD modeled organophosphorous pesticide (OP) mixtures in adult and developing
    rats in conjunction with OPP,  as part of the OP Cumulative Assessment (NHEERL).
       o  Results demonstrated greater-than-additive responses to a mixture of 5 OPs, with
          effects exacerbated in the young.  The experimental design and statistical analysis
          used to determine the effects of mixtures has also been successfully applied to
          carbamates, pyrethroid insecticides, and thyroid disrupting chemicals.
•   Research on chemicals that modify the regulation of luteinizing hormone: This work has
    demonstrated that a toxicant-induced alteration in luteinizing hormone secretion is a common
    mode of action underlying altered reproductive function and other critical reproductive
    outcomes (NHEERL).
       o  ORD research has been used in the Cancer Risk Assessment of Atrazine and the
          Chlorotriazine Cumulative Risk Assessment.  It also identifies the hypothalamic-
          pituitary-gonadal axis  as an area of concern for OPPTS' endocrine disrupters
          screening and testing program, and eliminates some forms of cancer as outcomes of
          concern with respect to animal to human extrapolation.
•   Models and data were critical for risk assessments and regulatory decisions on the
    developmental effects of specific pesticides (e.g., chlorpyrifos)
    http://www.epa.gov/oppsrrdl/REDs/chlorpyrifos_ired.pdf  (NHEERL).
•   ORD researchers have provided extensive expertise and review of numerous IRIS (Integrate
    Risk Information System) and OPPTS Risk Assessments (e.g., acrylamide, perchlorate,
    thiourea, perfluorooctane sulfonate, perfluorooctanoic acid, perchlorate, 2,3,7,8-
    Tetrachlorodibenzo-P-Dioxin (TCDD) And Related Compounds) http://www.epa.gov/iris/
    (NHEERL).
•   In collaboration with OPPTS, ORD developed harmonized guidelines (for cross-agency use)
    for immunotoxicity testing, developmental immunotoxicity testing, and the Risk Assessment
    Guidelines for Immunotoxicity (NHEERL).

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•  ORD researchers provided the expertise needed for the review of the Draft OECD 426
   Developmental Neurotoxicity Guidelines (NHEERL).

Safe Pesticides/Safe Products - Research program in LTG 1 since 2003:
•  Transition of staff to a new program focused on development of high-throughput in vitro and
   alternative species methods and peer review of program goals (NHEERL).
•  Developed and supported the first Center for Alternatives to Animal Testing (CAAT)
   TestSmart Developmental Neurotoxicity (DNT) Workshop on alternative methods for
   developmental neurotoxicity testing
   http://caat.jhsph.edu/programs/workshops/testsmart/dnt/index.htm (NHEERL).
•  Established an in house fish colony and published a medium throughput test method for
   exposure offish embryos
   http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&dopt=Abstract?
   lus&list uids=16620995&query hl=7&itool=pubmed docsum (NHEERL).
•  Developed in vitro assays and strategic chemical selection in a systematic approach for
   prioritization of large chemical inventories within a defined toxicity pathway (NHEERL).
•  Delivered test version of software for rapid and efficient depiction of structure searchable
   metabolism pathways and associated bioassay data (NHEERL).
•  Provided extensive technical support and a wide range of research activities in support of the
   OPPT perfluorooctanoic acid (PFOA) Risk Assessment and Enforceable Consent Agreement
   (EGA) activities (http://www.epa.gov/oppt/pfoa/).  Extensive research on the toxicity and
   pharmacokinetics of PFOA and perfluorooctyl sulfonate (PFOS) was critical  for these
   activities and is still under way. Sampling and analytical methods have been developed and
   validated for characterizing PFOA and other PFAAs in numerous environmental and
   biological matrices.   Protocols have been developed and are being used to evaluate the
   potential degradation of selected fluoropolymer-based polymers in soil and sediments.  The
   biodegradability of the same polymer products will be evaluated using both long term high
   microbial population  exposures (OPPTS SCAS testing) and simulated aerobic wastewater
   treatment conditions (OECD Simulations testing).  Test protocols for both thermal generation
   of PFOA by fluoropolymers and accelerated aged article testing have been developed which
   have been adopted by the fluoropolymer industry (in principle).  Quantitative method(s) to
   analyze PFAAs using LC Q-TOF MS which will be used to assess PFAA concentrations for
   all experimental conditions and matrices were developed.  ORD's research is producing
   state-of-the-science results that inform OPPT regarding the potential risks associated with
   environmental exposures to PFAAs as they continue to negotiate with industry through the
   ECA process..  (NHEERL, NRMRL, NERL).
•  Completed a 24-month study assessing the effectiveness of deck sealants to reduce exposure
   to arsenic in CCA-treated wood. This research is part of a multi-year effort that supported
   OPP's risk assessment and risk management activities on chromated copper arsenate (CCA).
   First year preliminary results were released in early May 2005.  A report has been completed
   and is expected to be  finalized in the Fall 2006 (NRMRL).
•  Developed and/or evaluated test protocols for detecting pesticides regulated by the Safe
   Drinking Water Act (SOWA) or for pesticides on the SWDA's Contaminant  Candidate List
   (CCL) under consideration for regulation. A model has been developed, and validated

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through laboratory experiments, for characterizing the fate of OP pesticides and their
degradation products as they travel from natural source waters through conventional drinking
water treatment plants (protocol website). Data generated from the protocols was used in
OPP's draft cumulative risk assessment document on n-methyl carbamates in 2005 (NRMRL
and NERL).
Collaborating with NCI, NIEHS, and NIOSH on the Agricultural Health Study (AHS), a
prospective epidemiological study to quantify the cancer and non-cancer risks in the
agricultural community and to study the relationships between agricultural exposures and
disease. ORD conducted the AHS Pesticide Exposure Study (AHS PES) at a small subset of
the AHS farms in Iowa and North Carolina (-120 out of-89000) to assess farm applicator
exposures to 2,4-D or chlorpyrifos during a single agricultural pesticide application. The
study results are being used by NCI, NIEHS, NIOSH, and EPA to assess/refine exposure
classification procedures for the AHS epidemiological study and to better understand the key
factors influencing agricultural  pesticide exposures to farm applicators and their families.
The AHS PES results have been provided to NCI and NIEHS for updating the AHS
algorithms and future questionnaires. EPA is reviewing the study results to address questions
regarding the 2,4-D reregi strati on eligibility decision (NERL).
Established an ORD NMR-Metabolomics Research Center with supporting data bases, for
identifying toxicity pathways and prioritizing testing.  This center is supporting various
research programs being implemented within and outside ORD including the conazoles
assessment, PFOA and other PFAAs (LTG 1), and the small fish study (LTG 2).  Delivered
test version of software for rapid and efficient depiction of structure searchable metabolism
pathways and associated bioassay data. (NERL, NHEERL).
Completed research in support of the Agency's  Asbestos Research Plan
(http ://www.epa.gov/opptintr/asbestos/ and
http://www.epa.gov/asbestos/pubs/vaiframework.pdf).  Exposure research was initiated to
evaluate filter collection efficiencies for airborne asbestos for various filter media and filter
pore size.  A method for characterizing asbestos in soils has been developed and provided to
the regional asbestos site coordinators.  The Releasable Asbestos Field Sampler (RAFS) has
been developed to obtain asbestos fiber releasability data from soil for calculation of
emission factors. The RAFS unit was first used in cooperation with EPA Regions 9 and  10
where Regional scientists were  on site conducting activity tests while ORD worked alongside
them collecting  data with the RAFS unit.  Preliminary data, study results and draft final
report are expected by December 31, 2006. Developed and published dose-response
relationships using a fiber  dose database compiled from past ORD characterizations of
exposures associated with toxicity studies conducted by ORD and other research groups.
This supports the on going  IRIS reassessment of cancer risks associated with asbestos,.
(NERL, NRMRL, NNHEERL)
Demonstrated that several chiral pesticides and other pollutants are enantioselectively
transformed by microbes in various environmental matrices, with some that selectively
accumulate in fish being toxic.  A definitive feature article, "Probing the Enantioselectivity of
Chiral Pesticides" (Environmental Science and Technology, Jan 06) summarizes the research
and highlights the importance of considering chirality in product registration (NERL).
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•  Issued three Requests for Applications for proposals from non-profit institutions in the area
   of Computational Toxicology. Awarded a total of six grants and three co-operative
   agreements, two of which established research centers on environmental bioinformatics
   (NCER).

Long Term Goal 2

•  Made available ECOTOX Release 4.0 August 2006. The ECOTOX database provides single
   chemical toxicity information for aquatic and terrestrial life.  Enhancements include
   expanded taxonomic and chemical searching, and output directly to MS Excel format
   http ://cfpub. epa. gov/ecotox/ (NHEERL).
•  Expanded ICE AND ACE datasets with the fathead minnow database of acute and chronic
   toxicity of industrial organic chemicals, and wildlife toxicity. Beta version of web-ICE
   developed for wildlife http://bagel.epa.csc.com/ice/index.htm (NHEERL).
•  Developed a conceptual systems model as a basis for predicting effects of hypothalamic-
   pituitary-gonadal (HPG)-active toxicants with differing mechanisms of action (MOA) in
   small fish species http://www.epa.gov/med/ (NHEERL).
•  Established a consortium of ORD and STAR-awarded scientists to conduct integrated studies
   linking genomic responses in small fish to outcomes at individual and population levels
   http://www.epa.gov/med/ (NHEERL, NERL).
•  Developed preliminary guidance on the development, application, and interpretation of
   population models for ecological risk assessment of pesticides http://www.epa.gov/aed/
   (NHEERL).
•  Developed PATCH, a spatially explicit, individual-based, life history simulator designed to
   project populations of territorial terrestrial vertebrate species through time
   http://www.epa.gov/wed/pages/models/patch/patchmain.htm (NHEERL).
•  Developed databases, a framework for spatial analysis, and test protocols to determine effects
   of herbicides on non-target crop and native  plant species for terrestrial plant risk assessments
   http://www.epa.gov/wed/pages/proiects/PesticideResearchFlyer.pdf
•  Developed and  provided to OPP a series of  ecological exposure models
   (PRZM/EXAMS/AgDrift) and accompanying databases in a probabilistic analysis
   framework (Express) for use in generating data required by FQPA
   (http://www.epa.gov/oppefedl/models/water/index.htm) and for the registration and/or
   reregi strati on of pesticides (http ://www. epa. gov/ceampubl/swater/express/index.htm)
   (NERL).

Long Term Goal 3

•  Developed methods for monitoring gene dispersal via pollen or seeds and probability of
   establishment of viable genetically modified (GM) plants or hybrids in natural areas
   http://www.epa.gov/wed (NHEERL).
•  Developed methods for genomic response (e.g., plant microarrays) and changes in fitness
   characteristics of plants with GM material as inputs for ecological risk assessments
   http://www.epa.gov/wed (NHEERL).

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Conducted in-depth review of scientific literature on genetic methods for long-term
ecosystem monitoring, implemented a pilot genetic monitoring program for ground beetle
communities, and coordinated a joint ORD/OPP workshop to explore ways to incorporate
these methods into agroecosystem assessment and monitoring (NERL).
Delivered a report on the genetic architecture of western corn rootworm, a target pest for new
varieties of Bt-corn. Successfully selected for Bt resistant populations, assessed gene flow
pattern across the USA, described the molecular structure of a cadherin-like protein that is a
candidate resistance gene, and developed a PCR assay to precisely monitor and detect
localized changes in the frequency of alleles at this gene (NERL).
Issued one Request for Applications for proposals from non-profit institutions in the area of
developing models for allergenicity in support of the biotechnology research program.
Awarded a total of four grants (NCER).
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                                  APPENDIX VII
                                   ACRONYMS
ACC         American Chemistry Council
ACE         Acute to Chronic Estimations
AGRA       Agricultural Biotechnology Risk Analysis
AHS         Agricultural Health Study
ALSase      Acetolactate Synthase
AOC         Articles of Commerce
APG         Annual Performance Goal
APM         Annual Performance Measure
ARS         Agriculture Research Service
ASTER      Assessment Tools for the Evaluation of Risk
HMD         Bench Mark Dose
BOSC        Board of Scientific Counselors
BRWG      Biotechnology Research Working Group
Bt           Bacillus  thuringiensis
CCA         Chromated Copper Arsenate
CEBRC      Carolina Environmental Bioinformatics Research Center
CENR        Committee on Environment and Natural Resources
CLA         CropLife America
CPRI         Crop Protection Research Institute
Comp Tox    Computational Toxicology
CPSC        Consumer Product Safety Commission
CTRP        Computational Toxicology Research Program
DNT         Developmental Neurotoxicity
DOD         Department of Defense
DQA         Data Quality Act
DW          Drinking Water
EAT         Estrogen, Androgen, and Thyroid
EGA         Enforceable Consent Agreement
EDCs        Endocrine Disrupting Chemicals
EDRP        Endocrine Disrupters Research Program
EFED        Ecological Fate and Effects Division
EPA         Environmental Protection Agency
EPRP        Ecological Protection Research Program
ERAs        Ecological Risk Assessments
EU          European Union
FBPP         Fluorotelomer-Based Polymer Products
FDA         Food and Drug Administration
FIFRA       Federal Insecticide, Fungicide, and Rodenticide Act
FQPA        Food Quality  Protection Act
FTE          Full-Time Employment Equivalent
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FTOH       Fluorotelomer Alcohol
FY          Fiscal Year
GE          Genetically Engineered
GIS          Geographical Information System
HED         Health Effects Division
HHRP       Human Health Research Program
HHRARP    Human Health Risk Assessment Research Program
HPA         Hypothalamic-Pituitary-Adrenal
HPG         Hypothalamic-Pituitary-Gonadal
HPV         High Production Volume
HPT         Hypothalamic-Pituitary-Thyroid
HTP         High-Throughput
ICCVAM    Interagency Coordinating Committee on Validation of Alternative Methods
ICE          Interspecies Correlation Estimations
ID           Identify
IgE, IgGl,    Immunoglobulins
IgA
IOAA       Immediate Office of the Assistant Administrator
IPCS         International Programme on Chemical Substances
IRM         Insect Resistance-Monitoring
KO          Knock Out
LC          Lethal Concentration
LD          Lethal Dose
LOAEL      Lowest Observed Adverse Effect Level
LRI          Long-Range Research Initiative
LTG         Long-Term Goal
LRAT       Long Range Atmospheric Transport
MOA        Mechanism/Mode of Action
MOE        Margin of Exposure
MYP         Multi-Year Plan
NCEA       National Center for Environmental Assessment
NCER       National Center for Environmental Research
NCI          National Cancer Institute
NCT         National Center for Toxicogenomics
NERL       National Exposure Research Laboratory
NHEERL    National Health and Environmental Effects Research Laboratory
NIEHS       National Institute of Environmental Health Sciences
NJRCEBCT  New Jersey Research  Center for Environmental Bioinformatics and
             Computational Toxicology
NMR        Nuclear Magnetic Resonance
NOAEL      No Observed Adverse Effect Level
NPD         National Program Director
NRC         National Research Council
NRMRL      National Risk Management Research Laboratory
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NTP         National Toxicology Program
OAR        Office of Air and Radiation
OECD       Organization for Economic Cooperation and Development
OIA         Office of International Activities
OMB        Office of Management and Budget
OPP         Office of Pesticides Programs
OPPT        Office of Pollution Prevention and Toxics
OPPTS       Office of Prevention, Pesticides and Toxic Substances
ORD        Office of Research and Development
ORMA       Office of Resources Management and Administration
OSCP        Office of Science Coordination and Policy
OSP         Office of Science Policy
OSTP        Office of Science and Technology Policy
OW         Office of Water
PART        Program Assessment Rating Tool
PATCH      Program to Assist in Tracking Critical Habitat
Pb           Lead
PBTK        Physiologically Based Toxicokinetic
PCBs        Polychlorinated Biphenyls
PCR         Polymerase Chain Reaction
PFAA        Perfluoroalkyl Acids
PFCs        Perfluorinated Chemicals
PFOA        Perfluorooctanoic acid
PFOS        Perfluorooctyl sulfonate
PIPs         Plant Incorporated Protectants
PMN        Pre-Manufacture Notice
PPA         Pollution Prevention Act
PPAR        Peroxisome Proliferator Activated Receptor
PTRP        Pesticides and Toxics Research Program
QSAR       Quantitative Structure Activity Relationship
R&D        Research and Development
RFA         Request for Applications
RP           Research Program
R&R        Renovation & repair
SBIR        Small Business Innovation Research
SEP         Supplemental Environmental Projects
SP2         Safe Pesticides/Safe Products
SRP         Sustainability Research Program
STAR        Science to Achieve Results
S&T         Screening and Testing
TH          Thyroid Hormone
TSCA        Toxic Substances Control Act
USDA       US Department of Agriculture
USGS        US Geological Survey
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WCR        Western Corn Rootworm
WHO        World Health Organization
WRS        Wildlife Research Strategy
WW         Wastewater
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