BRIEFING BOOK
                November 1986
                 Compiled by
                S. Mark Meyers
Environmental  Research Laboratory -- Corvallis
           Toxics/Pesticides Branch
              Corvallis, Oregon

                               Table of Contents
Introduction                                                                 1
     Overview                                                                2
     Goals and Objectives of the Wildlife Toxicology Team                    2
     Technical needs of the Agency                                           4
Accomplishments                                                              8
Current Research                                                            11
Facilities                                                                  15
     Avian Toxicology Facility (Greenhouse 8)                               17
     Wildlife Toxicology Facility (New Building)                            18
     Western Fish Toxicology Station (WFTS)                                 19
     WFTS Main Building                                                     20
Personnel                                                                   21
     Wildlife Toxicology Team                                               22
     Resume's                                                                23
          EPA                                                               23
          Visiting Scientists                                               31
          NSI                                                               34
A.   Experimental Synopses                                                  42
          Completed                                                         42
          Ongoing                                                           48
          Future                                                            57
     Extramural                                                             62
B.   Chemistry Capability and Laboratory Analysis                           70
C.   Publications                                                           73
     Inhouse                                                                74
     Extramural                                                             79
D.   Quality Assurance Program                                              82
E.   CERL Institutional Animal Care and Use Committee
     Interagency Research Animal Committee Proposed                         87


     This publication summarizes the goals and objectives of the Wildlife
Toxicology Research Team within the Toxics and Pesticides Branch of the
Corvallis Environmental  Research Laboratory (CERL).   In accord with the
objectives of the Agency, it is important to develop an in-house capability to
coordinate and conduct research in terrestrial wildlife toxicology.  This
capability serves several administrative and technical  functions, and provides
data needed to verify and evaluate effects of toxic  substances in support of
registration and use.
     Preliminary information about the biological  effects of toxic substances
can initially be answered in a laboratory environment.   Ultimately, however, it
is necessary to determine the relationship between standard EPA laboratory test
protocols and toxic effects observed in actual field exposures.  Only through
carefully coordinated laboratory and field investigations can this goal  be
     A primary goal of the Wildlife Toxicology Research Group is to compare
both laboratory and actual field toxicity data for chemicals of interest to
Office of Toxic Substances (OTS) and Office of Pesticide Programs (OPP).  Acute
toxicity and the coincident tissue residue levels  are determined in the labora-
tory and compared to those observed in the field,  to evaluate actual  field

     One of the primary  functions of the U.S. Environmental Protection Agency
is to evaluate the impact of chemicals and toxic substances on the environment.
Under the terms outlined in the Toxic Substances Control  Act (TSCA),  the EPA is
charged with the regulation of the manufacture, commercial  distribution, use,

and disposal  of chemical  substances and mixtures  shown  to  present  an  unreason-
able risk to human health or to the environment (Public Law  94-469, 1976).   In
response to this law,  the EPA must assess  the  magnitude of expected exposures
and the potential  effects of new chemicals (or of old chemicals  used  in  new
ways) on ecosystems and their components.   The toxic or deleterious effects  of
chemicals on biological systems is one of  the  most important considerations  in
determining the effects of a chemical  introduced  into an ecosystem.   An  eco-
system is a complex, stratified structure  which is made up of both biotic  and
abiotic components, and assessment of  the  impact  of a chemical introduced  into
an ecosystem should ultimately account for as  many interactions  of the system
as necessary.  The final  measure of the impact on an ecosystem is  the survival
of its components, but other subtle alterations in the  system may  be  less
dramatic.  The impact of some sublethal  effects are not obvious  until long
after the exposure occurs and thus may be  masked  by a significant  time delay.
     In the overall assessment of the  impact of any toxic  substances, it is
necessary to determine both the acute  (lethal) and subacute  (sublethal)  effects
on the species in  the exposed ecosystem.  The  impact of toxic substances on
terrestrial wildlife in an ecosystem is thus very complex.
     Concern about the effects of toxics on terrestrial  wildlife was  greatly
stimulated by the  passage of the Federal Insecticide, Fungicide, and  Rodenti-
cide Act (FIFRA),  which specifically and implicitly requires wildlife toxico-
logical testing.  In response to the FIFRA, guidelines  for the test and  evalua-
tion of toxic chemicals which impact terrestrial  wildlife  were published in  the
Federal Register on July 10, 1978.  The guidelines are  specific  and comprehen-
sive in scope, and form the basis for  most of  the current  interest in toxico-
logical testing.  The testing of potentially toxic substances under these
guidelines emphasizes the acute (short term, lethal) effects, as determined  by

LC50 and LC50 tests (FIFRA Section 163.71-1 and 163.71-2).   Less  understood  are
the chronic (sublethal) toxic effects.   Acute tests,  such as the  LD50  and LC50
tests, determine those levels of a chemical which,  when  ingested  (in a single
dose or in the diet), cause mortality in 50% of the treated animals.   LD50
tests provide data which can be used to estimate levels  of  chemical which, if
released into the environment, would produce significant effects  on the
animal s.
     Results from these rather basic laboratory tests cannot be easily used  to
predict or evaluate effects beyond the  single-species level  of organization.
Sublethal  effects of a chemical  may be  manifest in  the exposed population only
after significant elapsed time.   Thus,  in the review  or  registration of poten-
tially toxic substances, it is necessary to identify  both the acute and chronic

Technical  Needs of the Agency
     To refine the Agency's ability to  assess the potential  toxicity of chem-
icals, the Agnecy needs support from the CERL in several  technical and research
areas, including:
1.   Technical Assistance.  The Wildlife Toxicology Research Team provides
     technical assistance for questions pertaining  to effects of  toxic chem-
     icals in several scenarios, including interpretation of monitoring or test
     results, support for administrative decisions, and  evaluation of  long-term
     hazards (assessment).  The Wildlife Toxicology Research Team forms a
     nucleus of toxics expertise upon which the Agency can  draw for technical
     support in questions of registration or validation  of  guidelines.
2.   Test Protocol Guidelines.  The effort to validate test methods must be
     directed towards identification and modification of those procedures in

     the current guidelines which  can  produce  inconclusive or  spurious test
     results.   New (or modifications  to old)  tests must  be developed which
     account for the variations in exposure experienced  in natural  systems.
3.   Coordination of Comparative Toxicology Research.  Coordination is needed
     in the EPA research efforts in comparative  toxicology, especially in the
     context of comparing chemical  effects across terrestrial  wildlife species.
     The applicability of laboratory  test data to actual  field conditions and
     to wild species must be considered to determine whether or  not test
     results on selected laboratory species can  be used  to estimate toxicity in
     other species.
4.   Field Validation of Laboratory Test Protocols.  Toxicity  tests must be
     performed in actual field conditions to  determine the effects  of toxics on
     species in the field.  The field validation effort  provides direct compar-
     ison of chemical toxicity between laboratory tests  and in actual field
     conditions.  Coordination of  both laboratory and field test protocols
     provides synchronization of test data which allows  direct comparison of
     toxic effects between tests.   If the laboratory and field test protocols
     (and the subsequent data produced) are properly formatted,  the test
     results can be compared directly.
5.   Development of a Wildlife Toxicology Data Base.  An important  function in
     the Terrestrial Wildlife Toxicology Research Team is to coordinate and
     develop a comparative data base  using available research  and test data
     relating toxicity of chemicals to terrestrial wildlife.   These data
     include results from different types of  tests, effects on different
     species, relationships between single-species and multiple-species
     toxicity tests, microcosms, and  natural  ecosystem (field) tests.  This
     data base serves two important functions;  (a) it provides  a central

information storage and retrieval  network which can be used to evaluate
existing test results and to design new experimental  protocols;  and (b)
the data base provides a means of assessing toxicity data quickly and
efficiently for evaluations of structure-activity relationships (SAR) and
common mechanisms of action for selected classes of chemicals.  The data
base will become the central repository for all EPA terrestrial  wildlife
toxicology data and will be a valuable tool in the assessment of effects
of specific chemicals and classes of chemicals.  In addition, the data
base can be used to determine inter- and intra-species responses to
selected chemicals, and provides a valuable tool in the development of
computer simulation models of toxic effects of chemicals on physiological
systems for risk assessment.
Development of Sublethal Tests.  Toxicity tests are conducted in-house to
validate acute and chronic effects of chemicals on terrestrial wildlife.
Research in sublethal effects include relationships between body burden of
chemical and observed physiological/behavioral effects.  There are several
techniques which can be used to identify and measure sublethal effects,
especially effects on metabolic and enzymatic systems, and effects on the
immunological system.  Another important indicator of sublethal  toxicity
is evidenced in the decreased ability to survive stress and stressful
environments.  This is of particular importance to the continuation of any
species, since individual organisms have been shown to be more susceptible
to disease or other stresses following their exposure to environmental
contaminants.  Alteration in basal metabolic rates for wildlife exposed to
chemicals has also been demonstrated.  Under additional stressful condi-
tions, such reduced rates may result in altered behavioral  responses or
diminished energy available for food collection, and may lower the overall

     potential  for survival.   If  increased chemical  sensitivity is extrapolated
     to the population,  the  impact  of  stress  on a  system becomes very impor-
     tant.   Many simple  assays which are  good indicators of stress or trauma
     can be determined  from  blood or tissue samples.  Alterations in stress and
     other  physiological  responses  to  exposure of  toxic substances is an
     important element  of the  research program of  the Wildlife Toxicology
     Research Team at CERL.

     The laboratory thus  fulfills a functional role, but at the same time
provides the basis for  in-depth  research  into the  complex mechanisms and
interactions of toxic substances.



Developed a multiple regression technique for evaluating the response  of
birds to treated and untreated food in dietary choice  tests.  Birds  could
detect the presence of many chemicals on food at sublethal  levels, but
their response was affected by the abundance and availability of food
choices.  Results will be used to determine the potential  for birds  to
modify their exposure to chemicals by changing foraging patterns.

Demonstrated that short-term,  dietary exposure (400 ppm) to methyl
parathion can affect incubation behavior (nest attentiveness and abandon-
ment) and duckling production  in mallards.   Similar effects on nest
abandonment and reduced hatchling production have also been observed in
the field.  This test demonstrated the types of pesticide-related effects
which are not now considered in the avian reproduction test.

Developed a technique for evaluating egg shell  quality which is more
sensitive than the current egg shell  thickness measurement.  A test  using
bobwhites and mallards demonstrated a more sensitive  response to shell
breakage force than to thickness.

Determined that the accumulation of hexachlorobenzene  residue in bobwhite
egg yolks plateaus at approximately five times the dietary  concentration
(in ppm).

Determined that the toxic effects of organophosphate  chemicals to bobwhite
are more pronounced at lowered environmental  temperatures.   Decreasing the
test temperature from 35C to 28C decreases the LC50 of chlorpyrifos  from
590 ppm to 280 ppm.  This increased sensitivity to chemical at low temper-
atures indicates that exposure to toxic chemicals in  the field (where
environmental temperatures are often quite low) will  produce more mortal-
ity than indicated in standard laboratory tests.

Determined that the LC50 of chlorpyrifos to mallard ducks is more sensi-
tive to size (weight) than to  age.  Calculated LC50 values  varied by an
order of magnitude in birds of the same age,  but of different weights
(> 100 g difference).  This variation in size of birds can  result from
shipping or dietary differences.  This illustrates the importance of
selecting test birds of specific size and age.  Current OTS test guide-
lines allow birds from 10-17 days of age.  This range  can encompass  birds
with over 250 grams variation  in weight.

Demonstrated the importance of longer dietary tests (longer than 5 days)
for the chemicals which cause  delayed toxicity and those which produce
food avoidance.  Chemicals which cause delayed toxicity (e.g., anti-
coagulants) often show little  or no toxicity within the time frame of
standard test guidelines.  Many chemicals produce food avoidance and some
animals can survive the 5 days of dietary exposure (LC50) by not eating.
The toxicity of the test chemical is thus not appropriately measured.
Longer-term tests alleviate these confounding problems.

Determine that the accumulation of hexachlorobenzene  residue  in  egg yolks
of bobwhite hens fed HCB plateaus to approximately  five  times the  dietary
concentration within 28 days.   Accumulation  of  HCB  in eggs  is in addition
to the accumulation of chemical  in liver,  fat,  and  other body tissues of
the hens.  This relationship will  be used  to evaluate the capability of
estimating hen dietary uptake  from egg residues.

The reproductive impact of an  organophosphate on  nesting mallards  was
evaluated in both a standard laboratory test and  a  natural  (pond)  setting.
In contrast to the caged birds in the laboratory  setting, the birds on the
ponds were able to build nests and incubate  eggs.   These conditions
provided a more realistic test scenario to evaluate the  impact of  chemi-
cals on "natural" reproductive processes.  Treated  birds in both environ-
ments exhibited reproductive impairments typical  of OP exposure.  Similar
results in both environments illustrates the potential utility of  pond
tests as a realistic link between laboratory testing  and field conditions.

Dr. John Emlen, population ecologist at Oregon  State  University, was
selected as a Visiting Distinguished Scientist  to spend  one year doing
population modeling in the Wildlife Toxicology  Research  Group.   He is
active in determination of Risk Assessment Models for EPA and is currently
leading the CERL Risk Assessment Team.

Dr. Robert Ringer, distinguished Avian Physiologist from Michigan  State
University, spent one year in  the Wildlife Toxicology Research Group
conducting avian researches.  Dr. Ringer was at CERL  as  a part of  a
cooperative agreement and continues collaborative research  with  the
wildlife toxicology scientists at CERL.


     A.    How does  an  animal's  behavioral response to chemicals modify  its
          subsequent exposure?
          1.    Dietary discrimination  tests  (R. Bennett)
               a.    Measurement of  detectability
               b.    Role  of choices in discrimination
          2.    Response of  valley quail  to contaminated habitats  (Crawford)
          3.    Response of  sage grouse to contaminated habitats (Hal ford, Blus)
     B.    How is chemical exposure  modified  through  the food  chain?
          1.    Secondary  exposure to mink and  ferrets (Ringer)
          2.    Dioxin  in  large  animals (Texas  A&M)
     C.    What are  the relative sensitivities  of  various  routes of exposure?
          1.    Oral  (food and/or water)  vs.  dermal toxicity (Williams/Cairns)
          2.    Inhalation vs. oral  toxicity  (Williams/Gile)
     D.    What period  in  the  nesting cycle is  most sensitive  to chemical
          exposure?  Effects  on reproductive success.
          1.    Time  of OP exposure  related to  nest success  (R. Bennett)
     E.    How does  the duration of  dietary chemical  exposure  affect  toxicity
          and sublethal endpoints?
          1.    Five  vs. twenty-eight day LC50  tests  (Ringer)
     F.    How do bioassay measures  of  exposure relate to  chemical availability
          in  the environment?
          1.    Acetylcholinesterase assays from alfalfa and potatoe  field
               studies (Blus, Halford)
          2.    Acetylcholinesterase assays from nesting habitat studies
          3.    Acetylcholinesterase assays from laboratory exposure  (Maguire,

     A.    What biologically  relevant endpoints could be refined or revised for
          better determination  of chemical effects on reproduction?
          1.    What parameters  or methods of the current reproductive tests
               need to  be  evaluated?
               a.    HCB reproductive test, bobwhite (Shiroyama)
          2.    Is  the current test  the most realistic treatment schedule for
               OPs and  carbamates (timing, duration, and route of exposure)?
               a.    Effects  of  short-term OP exposure on nest abandonment and
                    success  (R.  Bennett)
          3.    Do  AChE  inhibitors affect levels of reproductive hormones?
               a.    Prolactin/OP interactions (R. Bennett, J. Bennett)
          4.    What is  the relationship between chemical residues in the egg,
               dose, and duration of exposure?
               a.    HCB residue movement into eggs (Williams)
          5.    What is  the best measure of eggshell quality?
               a.    Shell  thickness vs. breaking strength  (J. Bennett)
          6.    Do  laboratory reproductive tests predict field effects?
               a.    Field  reproductive effects with waterfowl (Kendall)
          7.    What are the  best techniques for measuring  reproductive success
               in  passerines?
               a.    Nest box technique development (Kendall, Blus, and Halford)
               b.    Red-winged  blackbirds (Meyers)
     A.   What factors affect dietary  toxicity?
          1.    How do alternative  dietary  choices affect response?
               a.    Discrimination tests (R. Bennett)
          2.    How does duration of the test and age of bird affect response?

          a.   Five vs.  twenty-eight day  dietary  test  (Ringer)
     3.   How do environmental  stresses affect response?
          a.   Temperature effects on LC50/LD50 (Maguire,  Williams)
     4.   How does previous chemical  exposure  affect response?
          a.   Pre-exposure experiments (Williams,  Bennett)
     5.   What is the relationship between  amount of chemical ingested,
          consumption, food passage rate, and  response?
          a.   Dose tests (Bennett)

B.   Population Level Effects
     1.   What methods are best for measuring  chemical exposure  and
          population effects?
          a.   Methods for sage grouse in  Idaho (Blus, Hal ford)
          b.   Methods for valley quail (Crawford)
     2.   Can complex population effects  be predicted  with models  of
          population dynamics?
          a.   Valley quail  methodology/lab interactions  (Emlen)


     The Wildlife Toxicology Research Team conducts  investigations  using the
mallard duck and bobwhite quail  as the test species  in accord with  the guidance
suggested in the FIFRA Guidelines for acute toxicity test protocols (Section
163.70-1).  It is necessary to provide housing and test areas for animals used
in these investigations according to good labortory  practices (GLP) and the
general protocol guidelines of the FIFRA.  In addition, animal  housing will
conform to Public Laws 89-544, 91-579, and 94-279 (Animal Welfare Acts of 1966,
1970, and 1976, respectively) which requires strict  adherence to general
procedures for the well-being of test animals.
     The husbandry support and test cages for the experimental  animals are
situated in a sophisticated test facility of approximately 4,000 square feet
(page 18).  This facility is adequate for each of the approved toxicity labora-
tory tests such as the LD50, LC50, and other variations of these tests.  In
support of these investigations, a laboratory preparation area, a food prepara-
tion area, a residue analysis laboratory, and a data anaysis area are also
located within the test area.  Large-pen (field simulation) testing is con-
ducted at Western Fish Toxicology Station (WFTS, page 19), utilizing the
existing ponds to provide a more natural environment for testing.  Investiga-
tions at this site are conducted in conjunction with tests using similar
protocols in the laboratory.  The combination of laboratory tests and large-pen
simulations provide an important link in verifying the relationship between
laboratory data and field data.
     The test area at the WFTS site is large enough  to support additional
laboratory or test space as required.  A block building (approximately 3.0 m  x
3.7 m) which is located on the test area is partially equipped for  laboratory


                                                                              CAGE WASH AREA
PN         PJ       PN
            i   c
          »9   /  I
        • ~   .   n
                             «   n
                        SUPPORT AREA


U . ^L



^\ 	 tl

O ^
n i >
TTTT /*1
i L"^


bi/s ^~7
— j
O K71

                               WILDLIFE  TOXICOLOGY FACILITY
                                                                           5    0

                                                                     jjg	•«
                                                                                  UMTCD CTATCS CNVWOMiKMTM.
                                                                                 WESTERN FISH TOXICOLOGY STATION CWFT8)
                                                                                   ISIO S f Qia»H»l AM. C«f«

                                                                                           SECOND FLOOR
                                                                                       UFTS MAIN BUILDING
                                                                                        MAIN BUILDING
                                                                              WESTERN FISH TOXICOLOGY STATION
                                             FIRST FLOOR
                                         UFTS MAIN BUILDING



     EPA Personnel
      Team Leader
    Bill A. Williams
   Research Biologist

   Richard S. Bennett
   Research Ecologist

   Michael A. Cairns

  Stephen E. Dominguez

    Anne Fairbrother
   Research Biologist

      Jay D. Gile

   Tamotsu Shiroyama

       Simon Wray
     Student (SIS)

   Section Supervisor
  David W. Schmedding

     Jewel Bennett

      Ray Bentley
  Associate Scientist

    Douglas Bonfoey

     Melissa A. Fix
 Laboratory Technician

     S. Mark Meyers

   Susan M. Schiller
   Laboratory Analyst




                               Richard S.  Bennett
                         Research Ecologlst -- Wildlife

             Ph.D. — Iowa State University,  Animal  Ecology/Environmental  1983
              M.S.  -- Michigan State University,  Wildlife  Ecology

              B.S.  -- Michigan State University,  Zoology


Research Ecologist
Corvallis Environmental Research Laboratory
     Relationships between patterns of chemical  exposure and
     effects; effects of sublethal  chemical exposure on avian
     nesting success.

Research Associate, National  Research Council           1983-1985
Corvallis Environmental Research Laboratory
     Avian dietary toxicity testing;  ability of avian species to
     detect pesticide-treated food.
              Research Associate
              Iowa Cooperative Wildlife Research Unit
                   Snow goose winter and spring migratory ecology.
              Graduate Research Assistant
              Iowa State University
                   Effects of synthetic pyrethroids on field populations of
                   invertebrates and small  mammals.

              Graduate Assistant                                     1977-1979
              Michigan State University
                   Response of ring-necked  pheasants to pesticide-treated
                   preferred foods; land owner surveys.

              Research Technician                                         1976
              Michigan State University
                   Upland game bird population ecology and habitat management;
                   census technique development.

AFFILIATIONS  Society of Environmental  Toxicology and Chemistry
              The Wildlife Society
              The American Ornithologists'  Union
              American Society of Mammalogy

                               Michael  A.  Cairns

EDUCATION     M.S. — Oregon State University,  Fisheries                   1980

              B.S. -- San Jose State University, Zoology  (Wildlife)        1974

EXPERIENCE    Biologist                                           1985-Present
              Corvallis Environmental  Research  Laboratory
              Wildlife Toxicology Group
                   Water route of exposure of waterfowl  to toxicants;  age and
                   size effects on avian sensitivity to  pesticides.

              Biologist                                              1981-1985
              Corvallis Environmental  Research  Laboratory
              Toxic Sediments Group
                   Freshwater fish and invertebrate sediment toxicity  test
                   development; bioassessment of acute and chronic toxicity of
                   contaminated sediment.

              Biologist, Lab Technician                              1976-1981
              Corvallis Environmental  Research  Laboratory
              Freshwater Division
                   Freshwater toxicity testing; developed toxicity test based
                   on fish respiration measurements; methods development;
                   sublethal stress/fish health.

              Biological Aide                                        1974-1976
              Corvallis Environmental  Research  Laboratory
              National Eutrophication Survey
                   Freshwater algal assays; nutrient limitation studies.

              Widlife Conservation Aide                                   1974
              Oregon Department of Fish and Wildlife
              Research Division
                   Conducted angler interviews, creel  censuses, ear counts to
                   obtain data on trout and steel head movement and catch.

AFFILIATIONS  The Wildlife Society
              American Fisheries Society
              North American Benthological Society

                              Stephen E.  Dominguez

EDUCATION     B.S.  — Oregon State University,  Microbiology                1972

EXPERIENCE    Biologist                                           1986-Present
              Corvallis Environmental  Research  Laboratory
                   Effects of sublethal  chemical  exposure  on  avian physiology.

              Aquatic Biologist                                      1980-1986
              Corvallis Environmental  Research  Laboratory
                   Lethality of heavy metals, organic  chemicals,  and  hypoxia  to
                   freshwater fish and invertebrates.

              Biologist                                              1972-1979
              Corvallis Environmental  Research  Laboratory
                   Effects of sedimentation on  biotic  communities and coho
                   salmon production in streams;  assessment of the extent of
                   culturally accelerated eutrophication of lakes within  the  48
                   contiguous states (EPA's National Eutrophication Survey).

              Physical Science Aide                                  1969-1972
              Corvallis Environmental  Research  Laboratory
                   Limnological research  related  to  causes and effecs of

                   Anne Fair-brother
                  Research Biologist

 Ph.D. — University of Wisconsin,  Madison,  Veterinary

  M.S. — University of Wisconsin,  Madison,  Veterinary
D.V.M. — University of California,  Davis                    1980

  B.S. — University of California,  Davis,  Wildlife Ecology  1976

 Research Biologist                                  1986-Present
 Corvallis Environmental  Research Laboratory
      Determine effects of sublethal  chemical  exposure on avian
      physiological parameters,  such as blood  chemistries and
      immune competence.

 Graduate Research Assistant                            1980-1985
 University of Wisconsin, Madison
      Determined interactive effects of pathogens,  toxicants, and
      temperature on immune function and bioenergetics of deer
              Research Assistant
              University of California,  Davis
                   Ectoparasite reinfestation rates  on  deer mice.
                   cycles of numbers of  deer mouse ectoparasites.


              Teaching Assistant
              University of California,  Davis
              Department of Wildlife and Fisheries  Biology
                   Mammalogy laboratory, teaching field identification of North
                   American mammals.  Summer field  course for wildlife students
                   to learn field techniques.
              U.S. Forest Service,  Inyo National  Forest
                   Conducted a survey of native fauna  in White and Inyo
                   Mountains, California.

AFFILIATIONS  American Society of Mammalogists
              American Veterinary Medical  Association
              Association of Wildlife Veterinarians
              Wildlife Disease Association

                                  Jay D.  Gile

EDUCATION     Graduate Study — Oregon State University                    1983
                                Phys i ology/Toxi cology

              M.S. -- Oregon State University, Marine Biology              1971

              B.S. — Central  Washington  State University,  Biology        1969

EXPERIENCE    Toxicologist                                        1982-Present
              Corvallis Environmental Research Laboratory
              Toxics/Pesticides Branch
                   Development of mallard research facility.   Conduct of avian
                   toxicity tests in support of FIFRA and TSCA guidelines.
                   Project officer for extramural  wildlife  research  projects.
                   CERL Radiation Safety  Officer.

              Research Biologist                                     1973-1982
              Corvallis Environmental Research Laboratory
                   Development of Terrestrial  Microcosm  Chamber (TMC) for
                   evaluation of chemical transport and  fate.   Project Officer
                   for extramural chemical  transport and fate  projects.

              Research Biologist                                     1972-1973
              Research Triangle Park (EPA)
                   Synergistic effects of radiation and  heavy  metals on mammal-
                   ian reproduction.

              Research Biologist                                     1971-1972
              Twinbrook Research Laboratory (EPA)
                   Synergistic effects of radiation and  heavy  metals on the
                   hematopoidic system in rodents.

                               Tamotsu Shiroyama

EDUCATION     B.S. ~ Oregon State University,  Microbiology               1969

EXPERIENCE    Microbiologist                                      1984-Present
              Corvallis Environmental  Research  Laboratory
              Toxics/Pesticide Branch
              Wildlife Toxicology Program
                   Oversee and coordinate all  the activities in the avian
                   (bobwhite) facility.   Also  provide assistance to ongoing
                   research projects.

              Microbiologist                                         1982-1984
              Freshwater Division
              Corvallis Environmental  Research  Laboratory
                   Determined the LC50 responses of selected freshwater phyto-
                   planktons to organic  toxicants that were used in ongoing
                   field and laboratory  microcosm studies.

                   Malheur National  Wildlife Refuge:   Studied the bioavail-
                   ability of nitrogen and phosphorus to phytoplankton.

                   Cooperative study on  "Textile Plant Wastewater Toxics
                   Study."  Provided toxicological  data on  freshwater algae
                   from wastewater samples collected from selected textile

                   Assisted in the development and refinement of the
                   Algal Assay Procedure:  Bottle Test.

                   Cooperative study with Western Washington State College on
                   biological field-laboratory  correlation  of primary producers.

                                Bill  A.  Williams
                               Research  Biologist

             Ph.D. — University of Illinois,  Neurophysiology

              M.S. — University of Illinois,  Physiology

              B.A. — University of California,  Physiology



              Research Biologist
              Corvallis Environmental  Research Laboratory
              Toxics/Pesticides Branch
                   Serves as project leader of Wildlife Toxicology Group at

              Research Scientist                                     1977-1982
              NASA-Ames Research Center
              Biosystems Division
                   Project Scientist in charge of biomedical  experiments for
                   Spacelab/Space Shuttle program.   Provided  scientific support
                   for zero-gravity biomedical experiments  for NASA.

              Research Physiologist                                  1972-1977
              NASA-Ames Research Center
              Environmental Research Center
              Environmental Control Research Branch
                   Project Scientist for NASA ground-based  biomedical  experi-
                   ments in temperature regulation  and metabolism.  Principle
                   investigator for metabolic experiments for Spacelab.  Served
                   as crewman for Spacelab missions  for NASA.

              Research Physiologist                                  1970-1972
              NASA-Ames Research Center
              Man-Machine Integration Branch
                   Research Scientist specializing  in  determination of chemical
                   interactions and control of temperature  metabolism and
                   hibernation in mammals.  Designed,  tested, and evaluated
                   liquidcooling techniques for astronaut space suits.

              National Academy of Science                            1969-1970
              Post-Doctoral Fellow
              NASA-Ames Research Center

              Instructor in Endocrinology                            1965-1966
              University of Illinois

              Instructor of Physiology                                    1965
              University of Illinois

AFFILIATIONS  Society of Environmental Toxicology and  Chemistry
              Sigma Xi
              American Physiological Society



Visiting Scientists

                               John Merritt Emlen
                                Senior Scientist

EDUCATION    Ph.D.  — University of Washington                             1966

              B.A.  — University of Wisconsin  (Madison)                    1961

              Haverford College                                      1956-1958

EXPERIENCE    Distinguished Visiting Senior Scientist              1985-Present
              Corvallis Environmental  Research  Laboratory

              Full  Professor (Courtesy)                           1984-Present
              Oregon State University
                   The evolution of patterns of population  density dependence
                   and interspecific interactions;  development  of  models  and
                   methods for determining density-dependence and  species
                   interactions in the field; modeling  of  impact of toxins on
                   spatial, dietary behavior.

              Associate Professor                                    1971-1984
              Indiana University
                   Trustee, Environmental  Defense Fund  (EOF).   Laboratory and
                   theoretical research on foraging and dietary behavior,
                   microhabitat associations,  reproductive  and  territorial
                   behavior, population genetics, and the  evolution of  ecolog-
                   ical characteristics; population consequences of life
                   history evolution.

              Assistant Professor                                    1966-1968
              State University of New York, Stony Brook

                                Robert K.  Ringer
Ph.D. — Rutgers University,  Physiology and Biochemistry     1955
              M.S. — Rutgers University,  Agriculture

              B.S. — Rutgers University,  Poultry Science

Department of Animal Science
Department of Physiology
Michigan State University
Coordinator of Toxicology, Pesticide Research Center
     Research Involvement: Effect of polybrominated biphenyl  and
     polychlorinated biphenyls on avian and mammalian species
     (mink and ferret), Refinement and development of wildlife
     toxicology protocols.  Effect of various toxicants on
     genesis of lymphoblasts.

Professor                                              1966-1978
Department of Poultry Science
Department of Physiology
Michigan State University
     Research Involvement:  Development of protocols for wildlife
     toxicology influence of polychlorinated and polybrominated
     biphenyls on avian and mammalian species; toxicology of
     methylmercury in mink; cardiovascular and reproductive
     physiology of domestic avian species.
              Department of Poultry Science,  Michigan State University

              Associate Professor                                    1961-1964
              Department of Poultry Science,  Michigan State University

              Assistant Professor                                    1957-1961
              Department of Poultry Science,  Michigan State University
              Assistant Professor
              Department of Poultry Science,  Rutgers University

AFFILIATIONS  Society of Environmental  Toxicology and Chemistry
              The Poultry Science Association
              American Physiological  Society
              American Association of Avian Pathologists
              World's Poultry Science Association



                   Jewel  Bennett

M.S. — Iowa State University, Wildlife Biology

B.S. — University of Wisconsin—Madison,  Biology


Northrop Services, Inc.
     Conducting research on the effects of environmental  contam-
     inants on avian reproduction; perform cholinesterase assays
     on avian tissue.

Wildlife Extension Writer                                   1982
Iowa State University
     Produced extension bulletins and magazine articles about
     Iowa's threatened and endangered wildlife.

Wildlife Research Assistant                            1978-1981
Iowa State University
     Conducted master's degree research on the seasonal range
     ecology of bison living in the mixed grass prairie at Wind
     Cave National Park, South Dakota.

Biological Aide                                        1979-1980
National Veterinary Services Lab, USDA
     Produced and conducted quality control tests on diagnostic

Wildlife Technician                                    1976-1978
University of Wisconsin
     Field assistant on a white-tail deer study in Wisconsin and
     a songbird censusing study in Montana.
              The Wildlife Society

                                  Ray Bentley
                              Associate Scientist

EDUCATION     B.S. ~ Oregon State University,  Wildlife  Science            1980

EXPERIENCE    Associate Scientist                                 1984-Present
              Northrop Services,  Inc.
                   Assists in experiments  on bobwhite  involved with  environ-
                   mental  hazards risk assessment.   Conducts  LCsg  testing  and
                   in charge of test facility  preparation  and maintenance.
                   Responsible for quail colony production,dispersal,  and
                   maintenance.  Assists in data analysis, necropsy,  and tissue
                   preparation for subsequent  residue  analysis.  Maintains
                   colony  production records and QA records on computer disk.

              Biological Aide                                        1983-1984
              Oregon Department of Fish and Wildlife
                   Assisted in data collection  and  analysis for  Tillamook  field
                   office.  Conducted census and demographic  data  on  coastal
                   black-tailed deer herds. Conducted district  1984  waterfowl
                   inventory.  Conducted Creek  survey  and  scale  sampling of
                   district river systems.  Participated as recorder  during
                   public  elk workshop sessions.

              Culturist                                                   1983
              Anadromous,  Inc.
                   Assisted in general  fish hatchery duties related  to salmon
                   production including spawning, scale  sampling,  vaccination,
                   disease treatment, loading  smelts for release,  measurement
                   and control of 02 levels and H20 flow/temp.

              Biologist Aide (Volunteer)                                   1983
              Oregon Department of Fish and Wildlife
                   Assisted in marking, attachment  of  transmitters,  and subse-
                   quent monitoring of spring  released pheasants.

                               Douglas  B.  Bonfoey

EDUCATION     M.S.  — Michigan State University, Chemistry                  1973

              B.S.  -- Western Michigan  University,  Chemistry                1967

EXPERIENCE    Analytical  Chemist                                  1983-Present
              Corvallis Environmental Research  Laboratory
                   Analysis of avian tissue for trace  amounts  of pesticides.
                   Methods development  utilizing solvent extraction,  sample
                   cleanup, and capillary  column gas chromatography.   Computer-
                   ized data handling and  data  analysis.

              Analytical  Chemist                                     1981-1983
              University  of Utah Research  Institute
                   Analysis of water and soil  samples  for trace  amounts  of
                   pesticides and PCBs.  Analysis  of hazardous waste  samples.

              Research Chemist                                       1973-1976
              The Dow Chemical Company
                   Process research: Experimentally determined  the optimum
                   reaction conditions  and purification methods  for a chemical
                   manufacturing process.   New  product research:  synthesized
                   and evaluated experimental  formulations.

              Analytical  Chemist                                     1968-1973
              The Dow Chemical Company
                   Provided analytical  chemistry support to  technical  service
                   groups.  Analyzed chemical  formulations using infrared
                   spectroscopy, gas chromatography, thin layer  chromatography,
                   UV-VIS and wet methods.

                                 Melissa  A.  Fix
                             Laboratory Technician

EDUCATION     A.S.  — Linn-Benton Community  College,  Liberal  Arts          1983

              A.S.  — Linn-Benton Community  College,  General  Science       1981

              Certificate -- Walter Reed  Research  Institute,  Animal Care
                             Specialist                                    1981

EXPERIENCE    Laboratory Technician                               1985-Present
              Northrop Services,  Inc.
                   Analysis of avian tissue, egg,  and fecal material  with  a  gas
                   chromatograph.  Assisting scientist in  data  and sample
                   collecting, necropsies, routine  animal  care,  data  charting
                   and entry.  Collect mallard blood  and run  PCV and  protein
                   analysis.  Run analysis of blood and tissue  on  SBA 300.

              Biological Aide                                             1985
              Corvallis Environmental  Research Laboratory
                   Analysis of avian egg  samples,  routine  animal care,  assist-
                   ing scientist  in measuring egg  shell thickness, literature
                   search, data entry and editing.

              Biological Aide                                        1984-1985
              Western Fish Toxicology Station
                   Routine care and analysis of  water microcosms measuring
                   frequency of microorganisms within microcosms.   Collecting
                   various pond waters.   Data entry and editing.

              Animal  Care Specialist                                 1981-1983
              U.S.  Army
                   Acting non-commission  officer of biomedical  facility.
                   Assisting in gastro-intestinal,  cardiac-pulmonary, and
                   reproductive surgeries.   Anesthetic and x-ray operator, pre
                   and post operative animal care,  necropsy,  tissue collection
                   and analysis.   Narcotic drug  control officer.

                   S. Mark Meyers
           Scientist (Wildlife Biologist)

M.S. -- Oregon State University,  Wildlife Science

B.S. — California State University,  Biological  Sciences

Scientist (Wildlife Biologist)                      1982-Present
Northrop Services, Inc.
     Conduct and support avian toxicological  research for the
     Wildlife Toxicology Program at CERL.   Development of a
     terrestrial toxicology data base (TERRE-TOX) for EPA, CERL.

Graduate Research Assistant                            1980-1982
Oregon State University
     Conducted field studies on survival,  reproduction, and
     habitat use of the ring-neck pheasant.  Completed Masters
     thesis and received M.S. degree from Oregon State University.

Biological Technician                                  1978-1979
Bureau of Land Management
     Recorded and evaluated physical and biotic stream character-
     istics.  Determined condition of riparian zones.  Evaluated
     human impacts on stream ecosystems.  Classified, rated, and
     inventoried public lands to determine suitability of habitat
     for wildlife.  Evaluated human impacts on wildlife habitats.

Biological Intern                                      1977-1978
California Department of Fish and Wildlife
     Assisted in the food habits study of the San Joaquin kit fox
     and bobcat.  Responsible for updating the reference
     herbarium for the Food Habits Laboratory.

The Wildlife Society
American Ornithologists' Union
Northwest Scientific Association


                 Susan M. Schiller
                 Laboratory Analyst

B.A. ~ Oregon State University,  German

Laboratory Analyst
Northrop Services, Inc.
     In support of the Wildlife Toxicology Group,  EPA.   Performs
     cholinesterase assays on avian tissue using Perkin-Elmer
     spectrophotometer or Gilford SBA 300.  Provide general
     biochemistry and chemistry support.   Assist in necropsies
     and collect data in support of eggshell  quality research.

Laboratory Analyst                                     1982-1985
Northrop Services, Inc.
     In support of the development of a wildlife toxicology
     database (TERRE-TOX).  Major duties include the acquisition
     and archiving of literature, quality assurance, information
     dissemination, and data entry.

Library Technician                                     1979-1981
Northrop Services, Inc.
     In support of the Corvallis Environmental  Research Labora-
     tory, EPA.  Responsible for literature acquisition, search-
     ing, recordkeeping, current awareness.

Library Aide                                           1976-1977
Oregon State University
Wililam Jasper Kerr Library
     In support of Business Records Department, processed contin-
     uations, typed vouchers, ordered missing journal  issues,
     general office work.
Phi Kappa Phi

                              David W.  Schmedding
                               Analytical  Chemist

EDUCATION     B.A.  ~ California State  University,  Chemistry               1969

EXPERIENCE    Analytical  Chemist                                  1983-Present
              Northrop Services, Inc.
                   Presently involved  in analytical  methods development  and
                   feed preparation and treatment  level  verification  for
                   toxicants used in IC^Q,  LDsQ, induced tolerance, and  repro-
                   ductive tests both  at CERL  and  WFTS.

              Research Chemist                                       1970-1983
              Department of Agricultural Chemistry

              Oregon State University
                   With a variety of pesticide and industrial  chemical research
                   projects.  Areas of  research include  nuclear magnetic
                   resonance studies on the binding of toxicants to biomem-
                   branes, physical  property determinations for chemicals  of
                   environmental concern,  determination  of adsorption isotherms
                   for various PCBs and pesticides on  sand, soil,  and clay
                   surfaces, and correlations  of physical properties  with
                   bioaccumulation.  Developed a method  to measure the evapora-
                   tive loss rate of aerial  agricultural  sprays as modified by
                   various adjuvants.   Work with toxicants led to an  involve-
                   ment in a U.S.-USSR  joint research  project  on substituted
                   aniline compounds and their mammalian toxicity as  related to
                   physical constants.   A cooperative  research project with the
                   University of Miami  School  of Medicine led  to the  solution
                   of the delayed neurotoxicological  symptomology and resultant
                   deaths of victims of accidental  ingestions  of high partition
                   coefficient organophosphate insecticides.   The interest in
                   organophosphates led to the development of  a Knudsen  effusion
                   apparatus for determining vapor pressures of "non-volatile"
                   compounds of interest for use in environmental  models on

              Research Chemist                                       1969-1970
              Frideh Research
                   Where work involved  computerized formulation and preselec-
                   tion of fluorescent  inks, intrinsic viscosity,  UV  quenching,
                   and volatility determinations with  subsequent plant produc-
                   tion and screening.




Title:  Effects of Adult Mallard Age on Avian Reproductive Tests

Introduction;  Avian reproduction tests are included in the hazard evaluation
process under the Toxic Substances Control  Act (TSCA).   Recommended test
species are the bobwhite (Cplinus virglnianus) and the  mallard (Anas
platyrhynchos).  TSCA guidelines specify several  parameters for selection  of
test birds:(1) the test birds should be pen reared either in the test labora-
tory or purchased from a commercial breeder; (2)  the source should have known
breeding history; (3) the sources of the test birds should be outbred with
genetically wild stock to maintain the genetic heterogeneity of wild birds;  (4)
the test birds should be phenotypically indistinguishable from wild birds.  The
guidelines further specify that the test birds be approaching their first
breeding season and at least 7 months old at the  beginning of the test.~After
these guidelines had been implemented, questions  arose  on the potential
variability of test results that might result from using young (7 month) birds.

Methods:  Two age groups of mallards were selected at the age extremes of  birds
approaching their first breeding season.  Birds 7 months old at test initiation
represented the youngest birds that EPA would accept in a reproduction test.
Birds 11 months old were used to represent the oldest birds which might be used
in this type of study.  The organophosphate insecticide, chlorpyrifos was
selected as the test chemical because of its widespread use on a variety of
crops and to compare with previous mallard reproductive tests conducted at the
Corvallis Environmental Research Laboratory (CERL).  Parameters measured
included adult food consumption and body weight,  brain  acetylcholinesterase,
onset of laying, egg production, fertility, hatchability, shell thickness, egg
weight, and duckling weight and survival.  Pairs  from each age group were
randomly assigned to reproductive pens housed in  a plastic covered greenhouse.
The birds were allowed to acclimate for 2 weeks,  then placed on the test diet
(80 ppm chlorpyrifos or corn oil control mixed with feed).  The birds were
weighed at 2-week intervals until the onset of laying and then at the end  of
the study (12 weeks after laying began).  Eggs were collected daily and set
weekly.  Upon hatching each duckling received a wing tag providing individual
identity and pen or parent identity.  The ducklings were weighed on day zero
and maintained.

Status:  Duckling day 14 weight was the only measured parameter which appeared
to be effected by adult age.  However, three of the 7-month old hens produced
atypically colored ducklings indicating a genotype different from the typical
mallard.  These ducklings were larger at day 14 than the other ducklings.   It
is possible that this phenotypical difference, although not evident in their
offsprings from all 7-month old hens, was not just restricted to the three pens
identified by the atypically colored ducklings.

     Since 11 of the 12 parameters tested were not affected by age, we conclude
that breeder age was probably not a factor in this test.

     This project was completed 12/85, and the manuscript has been accepted by
Archives of Environmental Contamination and Toxicology.

Investigators:  J. Gile, M. Meyers


Title:  Chlorpyrifos Toxicity to Bobwhite at Lowered Test Temperatures

Introduction:  It is necessary to determine the effect of environmental
stresses on susceptibility to chemicals in avians.Juvenile "homeothermic"
birds with developing thermoregularity capabilities may be adversely affected
by environmental  temperatures which are thermoneutral  to adults.  Such
responses may be exacerbated by pesticide exposure.  To evaluate the inter-
active effects of temperture stress and organophosphate exposure, 14-day-old
bobwhite quail were fed chlorpyrifos at four environment (test)  temperatures
(35, 32.5, 30, nad 27.5°C).  A standard LC50 (dietary) test design was followed
(test birds were fed dietary chlorpyrifos for five  days, followed by three days
of clean feed).  Results indicate that exposure of  the test  birds to lowered
environmental temperature during the dietary exposure to this organophosphate
produced LC50 values significantly lower than control  values (35°C test temper-
ature).  A significant (45%) drop in the LC50 occurred between 35°C (optimal
incubator temperature) and 27.5°C.  Brain cholinesterase (ChE) activity
measured in brains of non-surviving quail was significantly greater at low
temperatures.  This difference was as great as 70%  between 35 and 27.5°C.
Juveniel bobwhite exposed to chlorpyrifos were significantly more susceptible
to low test temperatures as evidenced by earlier mortality in the treated

Methods and Materials:  Juvenile bobwhite were exposed to dietary concentra-
tions of chlorpyrifos which bracketed the LC50 value for the 14-day-old
bobwhite at four test (environmental) temperatures.  The control birds were
housed at 35°C (optimal brooder temperature), while similar groups of birds
were tested at temperatures of 27.5, 30, and 32.5°C.  Values for the LC50  were
calculated using the probit method.  Results were reported as absolute LC50
values and as relative to control values.

Status:  The investigations completed were designed to evaluate  the sensitivity
of juvenile bobwhite in order to follow the standard test protocol.  These data
are representative of the effects of environmental  cold stress on young
bobwhite which are exposed to chemical.  Additional testing is planned to
evaluate the effects of cold stress on the toxicity of several organophosphate
chemicals in adult bobwhite.  Two manuscripts are in review for  submission to
Environmental Toxicology and Chemistry.

Investigators:  C. Maguire, B. Williams

Title:  Mallard Reproductive Testing in a Pond Environment:   A  Preliminary

Introduction:   The Toxic Substance Control  Act (TSCA)  and  the Federal  Insecti-
cide, Fungicide, and Rodenticide Act (FIFRA)  require  standard lethality  studies
(i.e., median lethal  dose, 1059, or median lethal  concentration,
be performed as part of the registration process  for new chemicals.   In  addi-
tion, TSCA and FIFRA have provisions which permit the U.S.  Environmental
Protection Agency (EPA) to require in-depth studies  into other potential
environmental hazards.   One such recommended study is a reproductive test
conducted with bobwhite quail  (Colinus virginianus)  and mallard duck (Anas
platyrhynchos).  Typically these studies are conducted in indoor laboratory
situations.  Th i s is desirable for both environmental control  and convenience,
but there is little evidence that these studies provide information  comparable
to exposure situations  in the wild.  A 2-year study  was initiated to determine
the feasibility of using outdoor pond enclosures  for reproductive testing on

Methods:  A 2-year reproductive study was conducted  on mallard ducks using 3-5
0.1 ha fenced ponds.   Ducks received feed treated with 0, 8,  and 80  ppm  chlor-
pyrifos for 18 weeks in 1984 and 20 weeks in 1985.  Opportunity was  given to
each hen to nest, incubate, and hatch out broods  with minimal  disturbance from
investigators.  The study was terminated for both years when  the last clutch on
each pond hatched and ducklings were 7 days old.

Results:  No negative effects were observed for mallards receiving 8 ppm
chlorpyrifos in their diet.  Birds receiving 80 ppm  chlorpyrifos hatched
significantly (p < 0.05) fewer ducklings per successful nest  (5.8) than
controls (10.2).  None  of the ducklings on treatment ponds  survived  to 7 days.
Control birds produced  8.4 ducklings per successful  nest surviving to 7  days or
longer.  Birds in the 80 ppm treatment group consumed less  feed than did
controls (p < 0.01).   Weight loss from reduced feed  consumption did  not  occur
to the extent we expected, indicating that birds  supplemented  their  diets with
natural foods found in  and around the ponds.  In  spite of relatively low
treated feed consumption, brain acetylcholinesterase was significantly (p <
0.05) depressed (57% of controls) for 80 ppm treated birds.  Studies on  indoor
penned mallards fed 80 ppm chlorpyrifos in their  diet also  resulted  in acetyl-
cholinesterase depression to the same extent, but at much higher feed consump-
tion levels.  The study described in this paper demonstrated  the potential of
using outdoor pond enclosures to evaluate chemical effects  on  food consumption,
brain acetylcholinesterase, and reproduction in mallards.  Furthermore,  repro-
ductive effects could be observed during a much shorter treatment period than
currently required for avian reproductive testing (approximately 20  weeks).

Status:  This project was completed 12/85, and the manuscript  has been pub-
lished in the Archives of Environmental Contamination and Toxicology

Investigators:  M. Meyers, J. Gile

Title:  Terre-Tox:  A Database for Effects of Anthropogenic  Substances  on
Terrestrial Animals

Introduction:   Terre-Tox is a new database developed for the U.S.  Environmental
Protection Agency to aid In Premanufacturing Notices and research.It  is
designed to become part of SPHERE (Scientific Parameters for Health  and Environ-
ment, Retrieval and Estimation) within the Chemical  Information System  (CIS).
Terre-Tox contains published (1970 to present)  information on the  toxicity  of
anthropogenic substances to terrestrial  animals.   It is comprised  of a  biblio-
graphic file and a data file.

Methods:  The bibliographic file is a listing of  all  articles collected,
reviewed, and accepted for Terre-Tox.  Articles on the effects of  chemicals on
birds, mammals, reptiles, terrestrial stages of amphibians,  and selected
invertebrates are collected.  These include laboratory, pen, and field  studies.

     After articles are obtained and entered into the bibliographic  file,
acceptable article are coded and entered into the data file.  The  material  in
the data file includes chemical information, test organism information, test
conditions, calculated endpoints (i.e.,  LDso, LCso),  and other results  (signs
of toxicity, behavior, avoidance, physiological,  etc.), and  comments.

Status and Results:  The bibliographic file contains 4,962 citations of pub-
lished articles from about 480 journals.  The data file consists of  test data
of more than 150 species and 800 chemicals.  At present there is a  total of
3,905 studies entered into the data file.   Both files can be word-searched  on
any of their fields.  The databases are  updated on a regular basis  as new
articles are obtained and coded.  A tape of Terre-Tox has been sent  to  NTIS
(National Technical Information Service).   It will  be available for  use by  the
general public (CSI, vendor).

Investigators:  M. Meyers, S. Schiller

Title:  Toxicity of Contaminated Prey to Carnivores

Introduction:  Previous studies have suggested there may be significant differ-
ences in mammalian carnivores in the toxicity of xenobiotics derived vTa
primary and secondary routes of contamination.  This study was designed to
assess the toxicity to carnivorous mammalian wildlife of selected potentially
hazardous xenobiotics derived via the consumption of contaminated prey.  The
mink was used as a test animal  representative of mammalian carnivores.  Compar-
ison of the toxicity of the metabolized forms of Aroclor 1254, Compound 1080,
Warfarin and Methrl Parathion (as contaminated natural  [live] food) were made
with comparable concentrations of the unmetabolized forms of the chemicals
added to the feed of the test species.


1.   Range-Finding Tests.  Each chemical was first fed to mink in a range-
find i?ig~TelT~fo~9etermTne appropriate concentrations for use in the definitive
tests (LCso).  Procedures for range-finding tests consisted of dosing via the
feed several mink with three to five widely-spaced concentrations of the
chemical.  A similar preliminary feeding trial with the prey species (rabbit)
was also conducted to ascertain the rate of bioaccumulation of the test sub-
stance in the whole body mass of these animals so that appropriate chemical
concentrations could be obtained, via the contaminated prey species, for
incorporation into the LC5Q test diets fed to the mink.

2.   Subacute Tests (LCsp).  A subacute (LCso) test was conducted with mink,
using proposed OTS mink toxicity guidelines, for each selected chemical.  The
LCso test consists of a 7-day acclimation period followed by a 28-day treatment
period.  One hundred twenty (120) mink were employed for each LCso test.  The
test animals were randomly assigned to cages.  In one test room, the animals in
the treatment groups were fed a basal diet in which the chemical being tested
was added to the diet in a geometric progression of concentrations designed to
produce from 10% to 90% mortality.  In another room, the diets fed to the test
animals in the treatment groups contained xenobiotic-contaminated prey, to
simulate secondary toxicity.  The concentrations of the test substance in the
diets of the treated animals in both rooms were comparable and all diets fed to
the animals in the controls and treatment groups in both rooms contained the
same percentage of prey species.  Feed and water were provided jid libitum
throughout the trial.

     Body weights of the test animals were recorded and reported at the initia-
tion of the acclimatization period, initiation of the test period, and at days
7, 14, 21, and 28 of the test period.  Food consumption was measured and
reported for all groups during the acclimation and test period.  Daily observa-
tions were made throughout the test period for any signs of intoxication or
other abnormal behavior and mortality.  At the termination of a test, LCso
values and 95% CI were determined by Probit analysis and comparisons made to
determine the comparative toxicity between primary and secondary exposure.

Status:  This project was requested by OTS.  All experimental work and analysis
is complete.  Journal articles have been submitted and a draft Secondary
Toxicity Test Protocol has been received by CERL.

Investigator:  R. Ringer

Ongoing Experiments

Title:  The Relationship Between Organophosphate Exposure,  Reproductive
Hormones, and Nesting Success

Introduction:  Research at CERL has shown that sublethal  exposure  of nesting
waterfowl to Organophosphate (OP) compounds may lead to reproductive failures
by causing nest abandonment and an abrupt loss of incubation behavior from
brooding hens.  There is evidence that OPs can cause a reduction in egg produc-
tion by disrupting the production or release of lutenizing  hormone (LH),  one of
the hormones responsible for ovulation.  This research was  initiated to deter-
mine if OPs also affect the release of prolactin, the hormone reponsible  for
incubation behavior in birds.  A disruption in the release  of prolactin in wild
incubating birds could lead to nest abandonment or reduced  nest attentiveness,
which would adversely impact hatchability and chick production.

     If the relationship between OP exposure and hormone  levels can be deter-
mined, it may be possible to screen for potential adverse effects  of chemicals
on reproduction with a blood plasma assay that determines the exposure level
responsible for disruption of egg production and incubation behavior.

Methods:  Preliminary research at CERL in 1985 exposed incubating  mallard hens
to methyl parathion-treated feed for 6 days.  Hens were allowed to set and
incubate their own clutches in indoor reproduction cages.  Blood serum samples
were collected routinely and analyzed for prolactin and LH.  The nest attentive-
ness of each hen was also determined with thermocouple probes that recorded the
temperature of each nest every 30 minutes.

     It was demonstrated, based on a small sample size, that sublethal methyl
parathion exposure to incubating hens produced a significant reduction in blood
serum prolactin.  Hens with prolactin depression also exhibited poor incubation
behavior as detected by a reduction in nest attentiveness.

     An expanded test was conducted in 1986 to further evaluate the relation-
ship between dietary OP exposure and reproductive hormones  as it affects
mallard nesting success.  This test monitored plasma prolactin concentrations,
plasma cholinesterase, blood chemistry parameters, corticosterone  levels, food
consumption, and nesting behavior throughout incubation for birds  in three
treatment groups:  control, 4000 ppm dietary methyl parathion, and paired-fed
groups.  The paired-fed group matched the reduced level of  daily food consump-
tion measured in the methyl parathion group.

Status:  Prolactin and blood chemistry assays are in progress.  The continua-
tion of this project in 1987 will be directed by the questions arising from
this year's findings, although further research into the  causes and ecological
importance of nest abandonment will continue in order to develop a better
capability of predicting chemical effects on nesting success.

Investigators:  R. Bennett, A. Fairbrother, J. Bennett, R.  Ringer, M. El-Halawani

Title:  Time Course and Level  of HCB Transfer From Hen  to  Egg  in  Bobwhite

Introduction:  It is necessary to determine the timing  and amount of dietary
HCB which reaches'eggs and embryos in aviansThexachlorobenzene  (HCB)  is  a
chlorine substituted aromatic  fungicide which accumulates  in body tissue and  is
transferred to the egg by laying hens.  The amount of HCB  in feed which is
necessary to produce transfer  and accumulation of chemical  in  bobwhite  quail
eggs has been determined.  Five levels of HCB were fed  to  paired  and isolated
laying bobwhite quail.  Total  egg HCB residue was measured for each  egg in
order to determine the time course of transfer of HCB from the food  to  the egg.
The total chemical level in food necessary to produce uptake of HCB  in  the egg
is related to levels which cause mortality (from LC50 data) and deficiencies  in
reproductive capability.  Total active ingredient consumed is  compared  to  egg
and hen liver residue at the end of the study to evaluate  the  exposure  to  HCB
which is necessary to produce  accumulation of chemical  in  eggs of bobwhite

Methods:  Adult laying bobwhite hens were exposed to dietary HCB  at  1.5, 3.0,
4.5, b.O, and 12 ppm and eggs  were collected for 48 days after the initial
exposure to chemical.  Egg yolks were separated and analyzed for  HCB residue
using a gas chromatograph.  Birds were placed back on clean feed  after  48  days
and eggs were analyzed for HCB.  Residues of livers were determined  in  some
hens at selected intervals and for all the remaining hens  at the  end of the

Status:  HCB residues have been traced into the eggs of bobwhite  hens at each
feeding levels of HCB in diet.  The residues begin to show up  in  significant
levels within 3 days (greater  than 1 ppm) and accumulate in egg yolks at a rate
of approximately 1.75 ppm/day.  After approximately 30  days of dietary  expo-
sure, both treatment levels of HCB reached plateau levels  approximately 5  times
(5x) the dietary concentration.  Research is continuing in an  effort to deter-
mine the movement of HCB residue into embryos of eggs produced by hens  fed
dietary HCB.

Investigators:  B. Williams, R. Ringer, T. Shiroyama, R. Bennett, Doug  Bonfoey

Title:  Egg Shell  Quality Evaluation

Introduction:  There is concern that environmental  contaminants may affect the
quality of egg shells produced by wild birds leading to egg breakage, nest
failures and, ultimately, effects on certain wildlife populations.  Conse-
quently, EPA requires that egg shell thickness be measured during reproduction
testing.  However, there are reports of chemical-associated egg breakage in
which significant shell thinning was not demonstrated.   Research at CERL found
many eggs produced by bobwhite dosed with sulfonilamide, a drug known to cause
shell thinning, retained normal thickness but were significantly weaker,
suggesting the ultrastructure of the shell  had been affected.

Approach and Methods:  Research is currently underway to evaluate a compression
test to determine egg shell strength as a potential additional  evaluator of egg
shell quality for EPA chemical registration tests.   A breaking strength test
may prove to be a method that can provide an especially relevant assessment of
the potential for a chemical to cause broken eggs in wild birds.

     The ultrastructure of the egg shells are also being examined by scanning
electron microscopy to determine if the chemical  exposure affects the shell
structure and how that relates to shell thickness and shell strength.

Status:  In 1986, two experiments were performed to compare the shell thick-
ness, shell strength, and ultrastucture of eggs produced by bobwhite exposed to
chemical substances.  One experiment involved a 9-week  exposure to DDE, the
stable breakdown product of DDT frequently linked to egg shell  thinning
problems in wild birds.  The other experiment exposed bobwhites to an organo-
phosphate insecticide for a 7-day exposure period.   The results of both experi-
mentes are currently being compiled and analyzed.  Scanning electron microscope
examination of shells produced in those experiments is  currently underway.

     Research in 1987 will compare the shell quality evaluation methods on
mallards exposed to various chemical substances.

Investigators:  J. Bennett, R. Ringer, R. Bennett,  B. Williams, P. Humphrey

Title;  Determination of the Ability of Avian Species to Discriminate Between
Treated and Untreated Food

Introduction:   One of the complicating factors in interpreting data from the
subacute dietary toxicity test (LC50) is the significant reduction in food
consumption associated with many chemicals at the higher dietary concentra-
tions.  For these chemicals the toxic effects are often confounded by chemic-
ally-induced starvation, which may or may not be indicative of the patterns of
field exposure.  Many avian species are able to detect and avoid chemically-
contaminated food in the lab when untreated or less toxic alternative food
exists.  But under what circumstances can birds respond to contaminated food in
the field by switching to less contaminated foods or feeding sites?  Our
research has shown that this ability is affected by the number of choices
available, the proportion of treated and untreated choices, and the detect-
ability of the chemical.  Other factors which will be investigated include
total  food abundance and physiological  status of the birds.

     The objectives of this ongoing project are:  (1) to determine the factors
influencing food avoidance behavior; and (2) to identify conditions under which
birds are able to respond to chemically-contaminated foods or habitats by
avoiding lethal or harmful exposure levels.  The results from this research
should aid in the interpretation of data from dietary toxicity tests, as well
as help to identify parameters that are important in realistic models of
chemical exposure for risk assessment.

Methods:  Thus far, the ability of bobwhite to detect and avoid food treated
with various pesticides has been determined in a series of food choice tests.
These tests consist of a 5-day treatment period followed by a 3-day recovery
period so that the results can be directly compared to the LC50 test.  The
tests range from a simple 2-choice test, with one food tray containing treated
food and the other untreated, to 10-choice tests where the proportion of
treated and untreated food trays can be varied.  The dietary concentration
above which birds exhibit a preference for the untreated food is defined as the
discrimination threshold (DT), and is calculated using a multiple regression
method developed at CERL.  An avoidance index can be calculated from LC50/DT
such that the higher the index value, the greater the potential to reduce
chemical exposure.

Status:  Laboratory tests have been conducted on 7 pesticides and manuscripts
are in preparation.  Further laboratory tests are planned to evaluate the role
of other factors on avoidance behavior and to test models of chemical exposure.
Future field studies are being planned to validate lab results and to further
identify field conditions under which chemical exposure is influenced by
behavioral responses.

Investigator:   R. Bennett

Title:   Normal  Mallard Blood Enzyme Values  for All  Age,  Sex,  and  Reproductive

Introduction:   Blood enzyme values are used routinely  in clinical  medicine  to
diagnose organ malfunctions.  It would be equally  useful  to use them  as  biolog-
ical markers indicating exposure to pesticides or  toxic  substances.   Unfortu-
nately, there is little information in the  literature  about normal values  in
mallard ducks, one of the avian species commonly used  in toxicological experi-
ments.   What information is available is applicable only to adult, nonbreeding
birds.   Since birds exhibit marked physiological changes during growth and
during  the reproductive period, it is necessary to generate normal values
throughout the mallard life cycle to evaluate  subsequent data on  enzyme  changes
following chemical exposure.

Methods and Materials:  Forty-eight adult mallard  pairs  will  be purchased  in
the spring in a nonreproductive state.  Birds  will  be  housed  in outdoor  ground
pens provided with nest boxes and materials.  Serum samples will  be collected
via jugular puncture at selected intervals  during  the  nonbreeding, egg laying,
and brooding periods.  Samples also will be collected  from chicks until  they
reach young adult age.  A panel of ten blood enzymes indicative of liver,
kidney, heart, pancrease, and muscle function  will  be  run, utilzing a Gilford
Impact 400 autoanalyzer, in cooperation with Dr. M. Craig, Oregon State  Uni-
versity School of Veterinary Medicine.

Status:  The experiment will be conducted from April 1986-March 1987.

Investigator:   A. Fairbrother, M. Craig, R. Bennett

Title:  The Role of Modeling in Risk Assessment at the Individual  and Popula-
tion Levels

Introduction:  The construction of models enables us to generalize from basic
principles to the predicting and understanding of novel situations.In the
case of chemical risks assessment, it makes possible the prediction  of a
specific chemical's impact on an ecosystem never before tested for sensitivity
to any chemical  at all.  Given the enormous array of species,  both chemical and
biological, models offer us the only hope for far-reaching,  reliable risk

Methods and Materials:  Models can be classified by types;  it  is our intention
to pursue three types:

(1) We intend to extend existing, species-specific models to incorporate the
effects of chemical insult.  Such models are not ideal because they  are general-
izeable only to the array of chemical, not biological  types.  They have the
advantage, however, of having been tested, are known to work,  and so fill a
short-term need for risk assessment in limited situations.

(2) We will construct "holistic" models driven by ecological principles and
empirically measured patterns that provide direct descriptions of chemical
effects on individuals or on populations.  Such modeling will  be extremely
flexible in that appropriate parameterization will permit application to almost
any chemical and biological species combination.

(3) We will construct "reductionist" models, based on first  (physical and
chemical) principles and general, ecological laws.  These models use chemical
uptake data and knowledge of physiology to predict toxic impacts on  indi-
viduals, or use toxicity information and laws of population  processes to
predict population-level impacts.  Unfortunately, because our  understanding of
toxicological and population processes are extremely poor,  such models, while
worth pursuing for their comprehensive, explanatory nature,  are unlikely to
bear fruit in the foreseeable future.

Status:  Underway.  Modeling effort will be coordinated with work done on
contract to Dr. Warren Porter and Ron Hindsell, University of  Wisconsin.

Investigators:  J. Emlen

Title:  Effects of an Acetylcholinesterase Inhibitor on  Red-Winged  Blackbird
(Agelaius phoem'ceus) Reproduction

Introduction;  To evaluate chemicals as potential  avian  toxicants,  the Environ-
mental Protection Agency requires chemical manufacturers to perform acute,
subacute, and/or reproductive tests on mallard ducks (Anas platyrhynchos)  and
bobwhite quail (Colinus virgianus).  In some cases the nng-necked  pheasant
(Phasianus colchicus) may be used.  As indicator species,  mallards, bobwhites,
and pheasants only represent about 5 percent of the total  avifauna  of North
America.  Passerine birds may constitute more than 50 percent of the total
post-breeding population. It appears apppropriate for future avian  toxicity
testing that a passerine bird(s) be included as an indicator (test) species in
TSCA and FIFRA guidelines.Research by the U.S. Fish and  Widlife Service  has
indicated that the red-winged blackbird is more sensitive  to toxic  chemicals
than other passerines tested and more sensitive than the mallard, quail, or
pheasant.  The red-winged blackbird is both abundant and ubiquitous in the
United States.  The objectives of our study are to determine the feasibility  of
using the Corvallis facilities (WFTS, duck ponds)  to conduct reproductive
studies on free-living birds and to determine the effects  of an acetylcholin-
esterase inhibitor on red-winged blackbird hatching process, fledgling, and
female behavior.  In addition, It is the goal of this project to determine  the
suitability of red-winged blackbirds as indicator species  as related to TSCA
and FIFRA guidelines.:

Methods:  Conduct a 2-year study using 6 0.1 ha fenced ponds at WFTS.  Each
pond is surrounded by a band of emergent vegetation that supports nesting
red-winged blackbirds.  Females will be captured and marked with color-coded
leg bands for future identification.  After a female completes her  clutch
(approx. 4 eggs) she will be captured and gavaged with either a carrier or
carrier plus test chemical in a random manner.  Female attentiveness to her
young will be monitored as well as hatching and fledgling  success.

Status:  The first season was completed in June 1986.  Methods were developed
for capture, tagging, and dosing blackbirds.  The second season will begin
April 1987 with the assistance of the Denver Wildlife Center (Ed Schafer and
John Cummings).

Investigators:  M. Meyers, E. Schafer, J. Gile

Title:  Changes in Mallard Hen Behaviors in Response to Methyl  Parathion-
Induced Illness of Chicks

Introduction:  Waterfowl chicks may become ill  following exposure to organo-
phosphates.  The hen must then choose to alter the behaviors of herself and the
remainder of the brood to accommodate the sick chicks or to maintain normal
behaviors and leave the sick young to follow as best they can.   If the former
course is taken, the hen may increase the risk of predation to  the entire brood
and decrease their ability to forage optimally.  If the latter  course if
followed, the sick chicks will most likely die.  Either way, the effects of
low-level exposure to the organophosphate may be different than predicted by
LC50 or LD50 trials.

Methods and Materials:  The methyl parathion LD50 for five-day  old ducklings
will be determined in a pilot study.  Hen and drake mallards will be housed in
ground pens with nest boxes and allowed to come into egg production and estab-
lish and incubate a clutch on their own schedule.  When ducklings in a brood
are five days old, half of them will be dosed by oral gavage with the pre-
determined dose of methyl parathion and the remainder will be dosed with
diluent (corn oil) only.  Numbered back tags will be put on all ducklings for
identification of individuals.  After dosing, the hen and her brood will be
released on one of the 4.9 meters deep ponds at WFTS and observed continuously
for 4 hours.  Observations will be made at selected intervals for the next 20
hours.  Observations will consist of noting the length of time  the hen spends
with the brood, foraging time, and behaviors of hen and ducklings, time spent on
land and on water, whether ducklings stay together in a group or if some spend
time alone, and any other behavioral observations considered significant.
After 24 hours postinoculation, broods and hens will be captured and removed
from the pond.

Status:  Small scale study was conducted in May-August 1986.  Expanded study is
under consideration for Spring-Summer 1987.

Investigators:  A. Fairbrother, M. Meyers, R. Bennett


Future Experiments

        Effects of Age on Mallard Sensitivity to Pesticides in  Avian  Dietary

Introduction:  Office of Toxic Substances (OTS)  guidelines for  the conduct of
avian dietary tests prescribe birds between ten  and seventeen days of age at
the start of exposure.  Office of Pesticide Programs (OPP) guidelines prescribe
ducks between five and ten days of age.   Literature perusal indicates that a
fairly significant variation in LC5Q values obtained from birds in this age
range (5-17 days) occurs for a given toxicant.  The standard test protocol may
need to be re-examined in terms of test organism procurement and age  at the
time of starting dietary exposures.  Differences in sensitivity at different
ages may be a function of size and not actual age.

Methods and Materials:  Mallards of at least three  different ages (i.e.,  5, 10,
14-d) will also be exposed to several  toxicants  using the standard protocol to
determine age-related differences in sensitivity.  To determine whether differ-
ences in sensitivity with age is due to development of detoxification mechan-
isms, liver enzymes may be measured as well.  These two experimental  strategies
should give insight into whether sensitivity is  age and/or size related.

Status:   The age sensitivity investigations began in the Summer of 1986 and
extend into and through 1987.

Investigators:   J. Gile, M. Meyers

Title:  Water as a Route of Exposure of Waterfowl  to Pesticides

Introduction:  Because the aquatic environment is  usually the eventual  recipi-
ent of most chemical contaminants, and because waterfowl  are so intimately
associated with that environment, we have initiated a study of waterfowl
exposure to toxicants via water.In initial  experiments  with mallard ducklings
exposed to methyl parathion in their drinking water, it was observed that
within 24 hours the concentration of chemical in the water column had decreased
to practically zero.  Our immediate objective is to determine the physical/
chemical fate of methyl  parathion, as well  as several other pesticides,  when
presented to mallard ducks via drinking water.  The longer-term objectives are:
(1) to experimentally determine water LCso's  for mallards exposed to various
pesticides; and (2) to experimentally determine the relative importance  of
water vs. food as routes of exposure to various pesticides.

Methods and Materials:  Physical/chemical  fate of  pesticides is to be examined
by short-term (24-48 hr) measurements of soluble chemical remaining with  and
without the presence of organic matter (i.e., feed, feces), as well  as  in
various types of water containers (i.e., glass, PVC, galvanized metal).   Water
LCso's will be determined by using a modification  of the  OTS Avian Dietary Test
(EG-15).  Relative importance of food vs. water as routes of exposure will  be
examined by exposing mallards to contaminated food, contaminated water,  both,
or clean food and water and statistically analyzing the resultant mortality,
growth, and cholinesterase activity using a factorial design.

Status:  Currently we are beginning to determine the chemical/physical  fate of
toxicants in the water matrix and will complete this investigation in FY87.  We
will also begin the determination of water  LCso's  for various pesticides  in
FY87 and continue in FY88.

Investigators:  M. Cairns, B. Williams, R.  Bennett

Title:  Immune Function Panel  for Avian Species

Introduction:   There are several immune function tests developed for avian
species, with  primary emphasis on the chicken.   The objective of this study
are:  (1) to determine which ones are feasible  for use with bobwhite quail and
mallard ducks; and (2) to select a panel  of tests that can be performed in
field laboratories with a minimum of specialized equipment yet still  provide
information about both cellular and humoral  immune functions.  Ultimately,
these tests will be used to determine if pesticides and toxic substances in the
environment cause immunosuppression in avian species,  making them more vulner-
able to endemic pathogens.

Methods and Materials:  Standard operating protocols gleaned from the litera-
ture or provided by other investigators will be followed,  making changes
necessary to adapt them to quail and ducks.   The following tests are being
considered for use:  macrophage migration inhibition;  delayed type hyper-
sensitivity; graft versus host reaction;  lymphocyte growth on chorioallantoic
membrane; natural killer cell  activity; lymphocyte blastogenesis; splenic
plaque forming assay (direct and indirect);  ELISA for  IgB  and IgM; total and
differential lymphocyte counts.

Status:   These experiments are currently in the planning stage.  Actual experi-
mentation should begin in the Fall of 1986.

Investigators:  A. Fairbrother

Title:  Effects of Environmental  Stresses on Avian Thermoregulation and

Introduction:  The objective of this test is to determine the effects of
environmental stress on the metabolism, thermoregulation, and sensitivity of
birds to toxic chemicals.  This project will provide information about the
ability of standard OTS toxicity tests to evaluate toxic effects in animals
subjected to stress conditions representative of those likely to be encountered
in the field.

Methods;  Standard OTS toxicity tests will be used to determine normal acute
toxicity values in bobwhite quail and the results compared with similar tests
in which environmental stresses are imposed.  Brain cholinesterase activity,
metabolic efficiency, and survivorship will  be monitored and evaluated in
relation to the interaction between chemical and temperature stresses.

Status:   This project will  begin summer of 1987 if funding is approved at an
appropriate level.

Investigators:  B. Williams, A. Fairbrother


Title:  Effects of Methyl  Parathion on Wild Avian Species in Agricultural  Areas
of Skagit Valley, Washington

Introduction:  The effects of organophosphate (OP) chemicals on  non-target
avians which utilize sprayed habitats needs to be evaluated.  This project was
designed to assess the effects of methyl  parathion (0,0-dimethyl  o,p-nitro-
phenyl phosphorothioate) on wild avian species in agricultural  areas of Skagit
Valley, Washington.  The study was designed to investigate the following two
objectives:  (1) to determine any disturbances in populations of non-target
avian species that are utilizing habitats that receive agricultural  spraying of
methyl parathion; and (2)  to assess reproductive and biochemical  (i.e77
cholinesterase inhibition) impacts of exposure of these bird species to methyl

     The production of teal and mallard broods and the survival  of teal  and
mallard ducklings were reduced by the application of methyl  parathion.   In
addition, four independent occurrences of abnormal bird behavior were observed
on the study fields following treatment with methyl  parathion.   These observa-
tions included abandonment of nests by mallard hens incubating eggs.  Other
researchers have noted disturbances in nesting behavior of birds exposed to
organophosphate pesticides.

     Occupancy of nest boxes by starlings was 73% on the treatment field and
82% on the control field.   Starlings began using the nest boxes  within one hour
after their placement in the fields.  Nest-building activity was evident in
many of the nest boxes within 24 hours.

Methods and Materials:  The experimental  design included the identification of
six agricultural areas that averaged approximately 28.3 hectares (70 acres) in
size and were approximately 60-70% bordered by waterways.  Three of these
treatment areas receive aerial application of methyl parathion at 0.7 kg/ha.
Data collected included:  (1) avian censusing; (2) carcass search; (3)  repro-
ductive study on starlings (Sturnus vulgaris); (4) residue analysis on environ-
mental samples (i.e., water, soil, and vegetation); and (5)  brain cholinester-
ase levels on birds.

Status:  This project was  completed in 1985 and several manuscripts are in
preparation.  (1) No significant difference was observed in the  number of young
starlings which fledged from nests on either field.  However, overall a lower
proportion of starling nestlings fledged from the methyl parathion treated
field (64.5%) when compared to the control field (89%).  (2) Hatchability of
starling eggs in exposed nest boxes was not significantly affected by applica-
tion of methyl parathion (1.4 kg AI/ha).  (3) Nestling mortality tended to be
higher on the treated field.  (4) Under the conditions of this study, aerial
application of 1.4 kg AI/ha methyl parathion did not affect reproduction in
breeding starling populations on the treated field.  Exposure via food chain
contamination was probably very low due to the apparent low density of insects
with associated low foraging activity on the field at the time of spray.

Investigators:  R. Kendall £t jiK, Western Washington University

Title:  Primary Hazards to Game Birds Associated with  the  Use  of Ramik  Brown
(Diphacinone Bait) for Controlling Voles in Orchards

Introduction;  EPA, Washington State Department of Game and Agriculture,  as
well as the U.S. Fish and Wildlife Service, have been  concerned about the
 otential hazards associated with rodenticides that have been  registered  in
 asrnngton to replace endrin use for controlling orchard mice.Preliminary
information indicated that chukar partridge may be more  susceptible than  quail
to anticoagulant poisoning (C. Henny, personal  communication).   Little is known
about the primary hazards of anticoagulants to  game birds under field condi-

     This study was conducted to evaluate the potential  primary hazards to game
birds resulting from the use of diphacinone bate to control  mice in orchards.
The two study areas were located in the Yakima  Valley in south-central
Washington, one was near Moxee, and the other near Sunnyside.

     In general, it appears that the diphacinone bait caused little if any
direct mortality to game birds.  However, pheasants and  quail  consumed large
quantities of the bait.  In some birds collected post-treatment, over 90% of
the crop contents was bait.  While these birds  appeared  normal, the long-term
effect of feeding on bait are unknown.  Some birds contained substantial
amounts of bait in their crop a month after treatment.

     While primary hazards appear to be low, secondary hazards may be a major
concern with the diphacinone bait.  Certainly the bait being consumed by  the
target and by game birds (even if they survive) presents a hazard to predators
that may consume them.  Some of the radio-equipped birds were killed and  at
least partially consumed by both mammalian and  avian predators.  In one case,  a
weasel had partially consumed a radio-equipped  quail.

Methods and Materials:  Sixty-eight wild ring-necked pheasants and 15 wild
California quail were captured, radio-equipped, and released in or adjacent to
orchards scheduled for treatment.  In addition, because  of difficulties in
capturing, 15 game farm-raised California quail and 41 game farm-raised chukar
partridge were released in or adjacent to orchards scheduled for treatment.
These 139 birds were radio-tracked before, during, and after the rodenticide
treatment.  Any birds found dead were necropsied and tissues were preserved
(frozen) for later residue analysis.  All birds with operational transmitters
at the end of the study were collected for necropsy and  tissues were preserved
for residue analysis.

Status:  Tests completed and final report and manuscript are in re-write  stage.
Anticipated publication as both an EPA document and a journal  article.

Investigators:  P. Hegdahl, U.S. Fish and Wildllife Service, Denver

Title:  Comparative Toxicity of Methyl  Parathion and Dieldrin in Laboratory
Rats and Mice, vs. Captive Feral Cotton Rats and White-Footed Deer Mice

Introduction:  It is often necessary to utilize laboratory animals to predict
hazard to terrestrial wildlife.  We are unclear, however,  as to the appropriate-
ness of using laboratory (surrogate) animals to extrapolate hazard data to wild
species.  This project was initiated to evaluate the similarities of response
of wild and laboratory mammals (rodents) to toxic chemicals.Acute oral  LDso
values for methyl parathion and dieldrin were determined in both sexes of four
rodent species.  Two laboratory species, albino rats and albino mice, and two
captive feral species, Sigmodon hispidus and Peromyscus maniculatus, were
chosen.  For methyl parathion, the males were found to be  more sensitive than
females and laboratory rodents more sensitive than captive feral rodents.  The
response of brain acetylcholinesterase (AChE) with respect to mortality was
also determined for the animals in the methyl parathion acute toxicity study.
A close agreement for brain AChE activity level was observed for both sexes in
both mice species.  A similar agreement was noticed for both sexes of both rat
species.  Brain acetylcholinesterase recovery (BAAR) was determined at day 28
for the feral rodent survivors, Sigmodon hispidus rats failed to recover,
whereas male Peromyscus maniculatus exhibited recovery and female Peromyscus
mam'culatus exhibited over-recovery when compared to brain AChE activity in
control animals.

Materials and Methods:  Sigmodon hispidus (cotton rats) and Peromyscus
maniculatus (white-footed deer mice) and domestic albino laboratory mice (Mus
musculatus) and the domestic albino laboratory rat (Rattus nprvegicus) were
compared in this study.  Acute oral toxicity of methyl parathion and dieldrin
was determined using standard oral gavage techniques.

Status:  The appearance of signs of intoxication, mortality, and apparent
recovery from intoxication were dose-dependent in the domestic rodents.  This
relationship was less apparent in the feral species.  Values for laboratory
rats and mice correlate closely with literature values.  The laboratory rodents
were found to be more sensitive (p = 0.05) to acute MP intoxication with the
exception of the male laboratory mice vs. male white-footed deer mice.

     This paper has been accepted for publication in Environmental Contamina-
tion and Toxicology.

Investigators:  D. Roberts, N. Silvy, E. Bailey, Texas A&M University

Title:  Uptake of 2,3,7,8-Tetrachlorodibenzo-p-dioxin by Dairy Cows

Introduction:  2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a highly toxic
byproduct formed during the commercial  preparation of chlorinated phenol -
derived products such as the phenoxy acid herbicides, and during the combustion
of diverse organics and municipal  waste.  Once in the environment, TCDD binds
tightly to the soil and usually remains on or near the soil  surface where it is
resistant to both photochemical and biochemical  degradation.  TCDD appears to
be particularly persistant in deeper and more protected soils with an estimated
half-life of more than 10 years.

     Absorption of TCDD from the gastrointestinal tract has  been extensively
studied in the rodent.  Relatively little is known regarding the fate of TCDD
in ruminants.  Recent studies suggest that the distribution  of TCDD in cattle
is similar to that seen in rodents with the liver or fat containing the highest
residue levels.  Given the highly toxic nature of TCDD. its  environmental
stability, its widespread occurrence as an environmental pollutant, and the
fact that grazing livestock are surely exposed to TCDD through incidental
consumption of soil and perhaps through other routes, it is  apparent tha"t a
thorough understanding of the interactions of TCDD with ruminants needs to be

Methods:  The research will focus on the following interrelated objectives:

(1) Isolation and identification of the metabolities of 2,3,7,8-TCDD in the

(2) Determination of the pharmacokinetics, metabolism and biological effects of
TCDD in the ruminant.  Both the parent compound and metabolites in the milk,
urine, feces, and various tissues will  be determined.  This  will permit the
comparison of the fate of non-bound TCDD with the effects of aged, soil bound
TCDD in the ruminant.

(3) Investigation of the bioavailability of TCDD from soil.   This study will
use a soil type to be approved by the EPA.  Cattle will be treated orally with
tritiated TCDD which has been allowed to age on the soil for four weeks.  The
level of TCDD exposure will be based on the best estimates of incidental  soil
consumption by grazing cattle.  Pharmacokinetic data, parent compound and
metabolite concentrations as well  as biological  effects on hepatic enzyme
activity will be determined.

Status:  This study is being conducted as a Cooperative Agreement with Texas A&M
University and is in response to a request by ORD.

Investigators:  Texas A&M

Title:  Relative Importance of Pathways of Exposure  and Influence of Environ-
mental Variables on Dose and Impacts of Parathion on Wild Game Birds

Introduction:  Pesticides'  impacts on wildlife, and  in particular wild game
birds, is an important issue associated with registration and permit review for
use of a chemical  in agriculture.  Data requirements include estimation of
effects on survival and reproductive potential  as well  as tissue residue
analysis from oral  laboratory feeding studies.   Other pathways of exposure,
such as;  (1) inhalation; (2) surface adsorption through feather and dermal1y
deposited pesticide; (3) indirect oral  ingestion through preening;  and (4T
percutaneous adsorption through the horny epithelial scales of the feetlmd
legs, have received little attention and are generally overlooked.

     Attempts to estimate relative importance of each pathway by simply expos-
ing wild game birds to spray of a pesticide in  animal aerosol exposure chambers
will not allow for accurate estimation of field exposures and potential
impacts.  Environmental variables, such as:  (1) time of day; (2) ground cover
type, density, and height;  (3) wind speed; (4)  temperature; and (5) relative
humidity influence the exposure concentration of pesticides reaching the bird.
Ground cover influences exposure by providing a physical shield for pesticide
interception along with a microclimate for the  birds.  Several of the other
environmental variables influence exposure by altering particle coagulation,
condensation and subsequently deposition velocity.  Additionally, time of day,
temperature, wind speed and relative humidity influence the physiologic state
of the bird which clearly influence the concentration necessary to produce a
given effect.  For example, the influence of temperature extremes on the bird's
metabolic rate could be the key variable that influences survival to a given

Methods:  The primary objective of this proposed two year research project will
be to provide a first order estimate of the relative importance of these
potential routes of exposure on dose (i.e., tissue concentration) and impacts
of the pesticide parathion on bobwhite quail.  Within the confines of the
experimental portion of this project we will determine the relative importance
of extremes of key environmental variables on both route of exposure and impact
of parathion to quail.

     During the first year of this study efforts will concentrate on examining
the relative importance of the routes of exposure, with limited time and funds
being focused on the influence of environmental variables.  Special emphasis
will be placed on examining inhalation of pesticides through nostrils, bronchi,
and into the alveoli of the lungs of the quail  because so little is known about
this route of exposure.  Then in the second year, if warranted, the focus would
be on the influence of critical environmental variables on exposure and
effects.  The final output of this project would be  a simple mathametical
computer simulation model that would allow one  to change certain key parameters
influencing concentration of exposure and estimate the dominant route of
exposure that needs to be examined in detail.  Parathion was selected for study
based on providing timely data for evaluating reregistration of this pesticide
as well as on the pesticide's physical  and chemical  characteristics.

Status:  This is a new project which will be conducted extramurally as an
interagency agreement with the U.S. Department  of Energy, Battelle Northwest.

Investigators:  Department of Energy

Title:  Risks of Chemical  Contamination to Mammalian  and Avian  Populations

Introduction:  Models have not yet been developed which assess  population level
effects of chemicals.The intent of this work is to  develop and test models
for predicting population-level impacts of chemical contamination on mammalian
and avian populations.

Methods and Materials:  Because of the vast array of  chemical and biological
species it is impossible,  by performing laboratory or field tests on a species-
and chemical-specific basis, ever to achieve a general  capability for chemical
risk assessment.  On the other hand, predictive population models, by allowing
us to extrapolate from general principles, permit the assessment of risks
arising from novel combinations of chemical and biological species.   We will
expand and develop population models, and gradually incorporate existing,
species-specific models into more general ones capable of predicting hazards
for a wide variety of situations.  Consideration will  be given  to avoidance
behavior, age (stage)-specific sensitivities, population density feedback,
inter-specific interactions (competitive and predative) and random fluctuations
of the natural, physical environment.

Status:  Several models have already been developed;  others are under consider-
ation.  Field studies for evaluating models of spatial  avoidance of toxicants
and the importance of species interactions to risk assessment have been
designed in detail for mammals, and are in an active  planning stage for birds.

Investigator(s):  TBD

Title:  Tracking of Avians in the Field Using Telemetry

Introduction:  The objectives of this task include:   (1)  to utilize telemetry
to develop the capability to accurately measure the  movements of quail  into and
out of fields which have been treated with pesticide; and (2) to document the
movement patterns of birds which reflect their ability to detect (and avoid)
chemically treated areas.  This project will  provide information about the
exposure of avians to chemically treated fields by documenting the movement of
avians into and out of the treated fields.

Methods and Materials:  The approach will  be  to set  up a  field test of sophisti-
cated radio telemetry tracking equipment and  to utilize wild quail trapped at
the 650-ha Wilson Game Farm south of Corvallis.  Each bird will  be tagged,
weighed, and equipped with a small radio transmitter coded to transmit a unique
signal.  The system will be automated to provide precise  time, location, and
movement information on each of the test birds.  The precise location and
movements of each bird will be determined and stored in the computer for
subsequent analysis of home ranges, habitats, breeding activities, and response
to chemical.

Status:  A cooperative agreement has been awarded to Oregon State University
for the conduct of this project.  A close scientific and  working relationship
with the Principal Investigator, Dr. John Crawford,  has begun. The first field
demonstration of this new technique will begin in early 1987.

Investigators:  J. Crawford, B. Williams, R.  Bennett



     The avian toxicology program has a requirement for feed and residue
analysis in the ongoing LC5Q, LDsg,  reproduction,  discrimination,  and induced
tolerance tests currently being conducted both  at  CERL  and  WFTS.  The ability
to verify toxicant concentrations in avian food allows  confirmation of levels
as well as meeting the required protocols of reproductive studies.  The track-
ing of toxicants and their metabolites permits  resolution of the more subtle
effects commonly occurring in a reproductive study.  The fate of ingested
chemicals varies with chemical  type.  They can  accumulate in the body, undergo
chemical breakdown (metabolize), be  rapidly eliminated  with no adverse effects
or, in the case of avian species, they may be eliminated through the egg  with
the resultant adverse effects on the offspring. Chemical  analysis of body
tissues aids the researcher designing studies to correlate  exposure with
toxicological effects.  After determination of  the body burden, it is possible
to estimate the dietary intake responsible for  those  toxicological effects.
Comparison of tissue residues and their effects found in laboratory tests can
be used to estimate the effects from actual field  exposures and design more
appropriate studies.

     The Wildlife Toxicology Research Team has  recently acquired a state  of  the
art HP 5880 dual capillary gas chromatograph outfitted  with several different
detectors to allow the quantitation  of many classes of  environmentally encount-
ered toxicants and their metabolites.  This instrument  is capable  of a high
degree of sample resolution at very  low levels  and, coupled with the high
sample capacity of the auto sampler, is a powerful analytical tool.

     Two additional gas chromatographs (HP 5830 and Varian  2700) are presently
used for backup and for methods development. A Waters  440  HPLC with an auto
sampler is used for high level  feed  analysis and for  heat labile compounds.
Also available for use within the branch are a  Varian 5000  HPLC, a Perkin-Elmer
2000 GC with auto sampler, and a new Finnigan 5100 GC/MS for chemical  variation
of unknown toxicants and their metabolites.


Blood Analyses

     One means of accurately determining the level  of  toxicity  exposure  of  an
animal is to measure several blood constituents  associated  with the  stress  or
toxic response.  Although several  new blood-related tests have  been  developed
in the last few years, initial  indications of stress are exhibited in  portions
of the differential blood count.   The lymphycote count can  be a direct indica-
tion of previous stress (e.g.,  the relationship  between "T" lymphocytes  and "B"
lymphocytes).  In addition to the  differential count and an analysis of  popula-
tion and types of cells present,  several clinical  enzyme and electrolyte assays
are now available which will identify recent stress-related responses.  The
tests are an important component of the CERL in-house  testing capability.   A
sophisticated Gilford SBA 300 Analyzer is on-line and  operational  for  these
tests.  A blood sample can also be used to determine cholinesterase  activity as
an indication of complications of anticholinesterases  or specific clotting  time
as an indication of complications  of anticoagulants.  The battery of tests
available from a blood sample and the relative ease by which it is obtained
make it a valuable tool in the assessment of the well-being of  test  animals.

Cholinesterase Determination

     The Wildlife Toxicology Research Team has developed standard  tests  for
acetylcholinesterase (AChE) activity in brain tissue and blood. These tests
are important because many pesticides (organophosphates and carbamates)  reduce
the AChE levels in the brain and blood resulting in dysfunction of nerve pulse
transmission.  Current methods include the use of the  Gilford SBA 300  and
Perkin-Elmer Lambda One Spectrophotometer, which are on-line at CERL.

Resi due Analysi s

     The analysis of chemical residue in selected tissues are determined using
GCMS and HPLC techniques.  These assays are important  to quantify  the  level of
toxic chemical exposure and the uptake of these  pesticides  in tissue after


                        Articles Published or Submitted

Aulerich, R. J., R. K.  Ringer,  and S.  Safronoff.   Assessment  of  Primary  vs.
     Secondary Toxicity of Aroclor 1254 to Mink.   Arch.  Environ.  Contain.
     Toxicol. (1986).   Submitted.
Bennett, R. S.  Role of Dietary Choices on the Ability of  Bobwhite  to  Discrim-
     inate Between Treated and  Untreated Foods.   Environ.  Toxicol.  Chem.
     (1986).  In press.
Bennett, R. S., and D.  W.  Schaefer.  Procedure for Evaluating the Potential  of
     Birds to Avoid Chemically-Contaminated  Food.   Environ. Toxicol. Chem.
Gile, J. D., J. B. Beavers, and R. Fink.  The Effect  of  Chemical  Carriers on
     Avian LCso Toxicity Tests.  Bull.  Environ. Contam.  Toxicol.  (1983).
Gile, J. D., and S. M.  Meyers.   Effect of Adult Mallard  Age on Avian Reproduc-
     tive Tests.  Arch. Environ. Contam. Toxicol.  (1986).   In press.
Kononen, D. W., J. R.  Hochstein, and R. K. Ringer. Avoidance Behavior of
     Mallards and Bobwhite Exposed to Carbofuran-Contaminated Food  and Water.
     Environ. Toxicol.  Chem. (1986).  Submitted.
Kononen, D. W., J. R.  Hochstein, and R. K. Ringer. A Graphical  Method for
     Evaluating Avian Food Avoidance Behavior. Environ. Toxicol. Chem.  (1986).
Meyers, S. M., and J.  D. Gile.   Mallard Reproductive  Testing  in  a Pond Environ-
     ment:  A Preliminary Study (1986).  V.  15:757-761.

Meyers, S. M., and S. M. Schiller.   Terre-Tox:   A Data  Base  for  Effects  of
     Anthropogenic Substances on Terrestrial  Animals.   J.  Chem.  Info.  Comput.
     Sci. (1986).  V. 26:33-36.

       Presented at the Fifth Annual  Meeting  of Society of Environmental
       Toxicology and Chemistry, Arlington, Virginia, November 4-7,  1984

Bennett, R. S.  Role of Dietary  Choices on Pesticide  Discrimination  in Bob-
     white.  Poster.
Gile, J. D.  Comparative Reproductive Effects of Chlorpyrifos on Mallard Ducks:
     Indoor Pens vs. Natural  Habitat.  Poster.
Maguire, C. C.  Cholinesterase Activity:   Depression  and Recovery Rates  in
     Bobwhite Quail Exposed to an Organophosphorus Insecticide.   Presentation.

       Presented at the Sixth Annual  Meeting  of Society of Environmental
      Toxicology and Chemistry,  St.  Louis, Missouri,  November 10-13,  1985

Bennett, J. K., R. K. Ringer, and B.  A. Williams.  Evaluation of Methods to
     Determine Eggshell Quality.  Presentation.
Bennett, R. S.  Comparison of Pesticide Effects on Food Avoidance Behavior in
     Bobwhite.  Presentation.
Gile, J. D., B. A. Williams,  and S.  M. Meyers.   The Dietary  Toxicity  of  Methyl
     Parathion to 14-Day-Old  Mallards.  Poster.
Maguire, C. C. and B. A. Williams.   Chlorpyrifos Toxicity  to Bobwhite  at
     Lowered Test Temperatures.   Poster.
Meyers, S. M. and S. M. Schiller.  Terre-Tox:  A New  Data  Base for Effects of
     Anthropogenic Substances on Terrestrial  Animals.   Poster.
                                       / O

Ringer, R. K.  The Future of a Mammalian  Wildlife  Toxicology Test.   Presenta-
Ringer, R. K., T. Shiroyama, and R.  Bentley.   Hexachlorobenzene on  Reproduction
     of Bobwhite.  Presentation.
Williams, B. A.  Wildlife Toxicology at CERL:   A Look  to  the Future.   Presenta-
Williams, B. A., R. K. Ringer, and T.  Shiroyama.   Time Course and Level of HCB
     Transfer from Hen to Egg in Bobwhite Quail.   Presentation.

    Presented at the Seventh Annual  Meetings  of the  Society of Environmental
       Toxicology and Chemistry, Alexandria,  Virginia, November 2-5,  1986

Bennett, R. S., J. K. Bennett, M. El Halawani, and R.  K.  Ringer.  Relationship
     Between Dietary Organophosphate Exposure, Prolactin  Levels, and Incubation
     Behavior in Mallards.  Poster.
Bennett, R. S., B. A. Williams, D. W.  Schmedding,  and  J.  K. Bennett.   Effects
     of Short-Term, Dietary Exposure to Methyl Parathion  on Nesting Success  in
     Mallards.  Presentation.
Williams, B. A., T. Shiroyama, D. Bonfoey,  and R.  Bentley.  Hexachlorobenzene
     Uptake in the Eggs and Embryos of Bobwhite.   Presentation.

                            Articles  in  Preparation

Bennett, J. K.,  R.  K.  Ringer,  R.  S. Bennett,  and B. A. Williams.  Comparison of
     Breaking Force and Shell  Thickness  of  Sulfanilamide-Influenced Bobwhite
Bennett, R. S.  Comparison of  Pesticide  Effects on Food Avoidance Behavior in
Bennett, R. S.,  B.  A.  Williams, J. K.  Bennett, and D. W. Schmedding.  Effects
     of Sublethal  Methyl  Parathion Exposure on Nesting Success  in Mallards.
Emlen, J. M.  Competition, Density Feedback,  and Dispersion Patterns in
     Atriplex confertifolia.
Emlen, J. M.  Interaction Assessment:  An Appplication and Test with Orzyopsis
Maguire, C. C. and B.  A.  Williams.  Acute Temperature Stress and Organophos-
     phate Effects on  Juvenile Bobwhite.
Maguire, C. C. and B.  A.  Williams.  Chlorpyrifos Toxicity to Bobwhite at
     Lowered Test Temperatures.
Williams, B. A., T. A. Shiroyama, and R. K. Ringer.  HCB Uptake in Egg and
     Embryo of Bobwhite.

                                In-House Report
Bennett, J. K. and B.  A.  Williams.  Acetylcholinesterase Determination Proce-
     dure.  Corvallis  Environmental Research  Laboratory, U.S. EPA (1985).
     28 pp.
Bennett, R. S. and D.  W.  Schafer.  Procedure  for Determining the Potential of
     Birds to Avoid Chemically Contaminated Food.

Bennett, R.  S.   Effect of Methylparathion  on  Mallard  Nesting  Success  in  Outdoor
     Pens.  1985.   4 pp.
Bennett, J.  K., and B. A. Williams.   Use of the  Assay for  Acetylcholinesterase
     to Determine  Exposure to Organophosphates or  Carbamates  in  the  Field.   EPA
     Special  Report.  1986.
Emlen, J. M., and  R. S. Bennett.   Review of Effects of Pesticides  on  Avian
     Foraging Strategies.  1985.   20 pp.
Ringer, R. K.  Protocol for Dietary  LC5Q Test to Assess Primary  and  Secondary
     Toxicity to Mammalian Wildlife.  Michigan State  University.   1986.   14  pp.

                       (Supported All  or in  Part  by CERL)

Berry, S. C.,  C.  J.  Driver,  R.  J. Kendall, and T.  E.  Lacher.  Effects of
     Agricultural Spraying of Methyl  Parathion on  Reproduction  and Cholin-
     esterase  Activity in Starlings (Sturnus vulgaris)  in Skagit Valley,
     Washington.   Sixth Annual  Meeting of Society  of  Environmental Toxicology
     and Chemistry,  St. Louis,  Missouri.  November 10-13, 1985.
Blus, L. J., D. K. Halford,  C.  J. Henny, T.  Craig, E. Craig,  and C. M. Bunck.
     Relations of Organophosphate Insecticides to  Sage  Grouse in Idaho.   56th
     Annual  Meetings of the  Cooper Ornithological  Society.  Davis, California.
     September 1986.
Brewer, L. and R. J. Kendall.  Effects of Agricultural  Spraying of Methyl
     Parathion on Nesting Ducks and Duck Broods in Skagit Valley, Washington.
     Sixth Annual Meeting of Society of Environmental Toxicology and Chemistry,
     St. Louis, Missouri.  November 10-13, 1985.
Galino, J. D., R. J. Kendall, C.  J. Driver,  and T. E. Lacher, Jr.  The Effect
     of Methyl Parathion on  the Susceptibility of  Bobwhite  quail (Colinus
     virgim'anus) to Domestic Cat Predation. Pacific Northwest Association of
     Toxicologists Meeting,  Seattle,  Washington.   September 1984.
Galino, J. D., R. J. Kendall, C.  J. Driver,  and T. E. Lacher.  The Effect of
     Methyl  Parathion on Susceptibility of Bobwhite Quail (Colinus virgim'anus)
     to Domestic Cat Predation.  Behavioral  and Neural  Biology  (1985).
Hill, E. F., and M.  B. Camardese.  Toxicity  of Anticholinesterase Insecticides
     to Birds:  Technical Grade Versus Granular Formulations.   Ecotoxicol.
     Environ.  Safety (1984).  V.8:551-563.

McAlpine, C., R. Kendall, C. J. Driver,  and R.  Thompson.   Effects  of  Methyl
     Parathion on Tonic Immobility and Brain Acetylcholinesterase  Activity  in
     Bobwhite Quail  (Colinus virgim'anus).   Pacific  Northwest  Association of
     Toxicologists Meeting, Seattle,  Washington.   September 1984.
Roberts, D. K., N. J.  Silvy, and E. M. Bailey.   Comparative Toxicity  of  Methyl
     Parathion and Dieldrin in Laboratory  Rats  and Mice vs. Captive Feral
     Cotton Rats and White-Footed Deer Mice. Arch.  Environ. Contam.  Toxicol.
     In press.
	, E. M.  Bailey, and N.  J.  Silvey.  Recovery  Patterns of Brain
     Acetylcholinesterase Activity Following Non-Lethal  Acute  Methyl  Parathion
     Intoxication in Sigmodon^ histidus,  Mus musculatus,  and Their  Laboratory
     Animal Counterparts.  Arch. Environ.  Contam.  Toxicol. (1986). Submitted.
	, E. M.  Bailey, and N.  J.  Silvey.  Comparative Acute Toxicity of
     Methyl Parathion and Dieldrin Between Three Indigenous  Texas  Rodent
     Species and Their Laboratory Animal  Counterparts.   Bull.  Environ.  Contam.
     Toxicol. (1986).  Submitted.
    	, E. M. Bailey, and N. J.  Silvey.   A Report of the Animal Hus-
     bandry Experiences in the Production of Colony-Reared Omnivorous  Rodents
     From Captive Feral Stock.  Lab. Anim. Med.  (1986).   Submitted.
    	, E. M. Bailey, and N. J. Silvey.   Brain Acetylcholineterase
     Activity Following Lethal  Methyl  Parathion Intoxication  in  Various  Captive
     Feral Colony-Reared and Laboratory Rodent Species.   Bull. Environ.  Contam.
     Toxicol. (1986).  Submitted.
    	, E. M. Bailey, and N. J. Silvey.  Use of  Colony-Reared Rodents
     From Captive Feral Stock as an Animal  Model  for the  Assessment of Environ-
     mental Pesticide Toxicity.  Ecotoxicol.  Environ. Safety (1986).   Submitted,


Roylance, K.  J.,  C.  D.  Jorgensen,  G. M. Booth, and M. W. Carter.  Effects of
     Dietary  Endrin  on  Reproduction of Mallard Ducks (Anas platyrhynchos).
     Arch. Environ.  Contam.  Toxicol.  (1985).  V.14:705-711.
Smith, D. E., and R. J.  Kendall.   Assessing the Effects of Pesticides on
     Wildlife.  J.  Pesticide Reform.  (1985). V.5:5-7.


                 OUTLINE OF


                  FOR THE

    U.S. Environmental Protection Agency

     Office of Research and Development

            200 S.W. 35th Street

          Corvallis, Oregon 97333

                               TABLE OF CONTENTS
     1.1  Introduction
     1.2  QA Policy Statement
     1.3  Laboratory Mission
     1.4  Administrative Sample Handling Procedures
     1.5  Data Generation  •
     1.6  Organism Care
     1.7  The Audit Program
     1.8  QA Responsibilities
     1.9  Resources for the QA Program
     1.10 Annual Planning
     1.11 Training

     2.1  Introduction
     2.2  Organization of the Corvallis Environmental  Research Laboratory
          Analytical Support
     2.3  Facilities, Equipment, and Support Services
     2.4  Maintenance Contract
     2.5  Automated Data Processing (ADP)

     3.1  Introduction
     3.2  Training
     3.3  Data Quality Objectives (DQOs)

     3.4  QA Project Plans
     3.5  Analytical Laboratory QA Plans
     3.6  QA Audits
     3.7  Peer Review
     3.8  Publications and Reports
     3.9  Performance Evaluation Program
     3.10 QA Resources
     3.11 QA Operating Constraints

Appendix A.  Guidelines for Development of Quality Assurance Project Plans
Appendix B.  Quality Assurance Review Form for CERL Extramural  Projects
Appendix C.  Quality Assurance in Organism Care and Testing
Appendix D.  Quality Assurance Project Review for the Corvallis Environmental
             Research Laboratory
Appendix E.  Laboratory QA Organization
Appendix F.  Research Notebook Policy
Appendix 6.  Annual Audit Summary Report for FY 1985
Appendix H.  Problems Discovered During Audits
Appendix I.  Peer Review Report Findings


(A Process of Obtaining Data of  Known Quality)

           Data Collection and Use

              Determine Criteria
                 for Decision
            Identify Data and Data
            Qualilty Requirements
               Design Study and
           Prepare QA Project Plan

            Develop Sampling Plan

              Choose Appropriate
              Analytical  Methods
           Perform Data Collection
             with Quality Control
            Assess and Report Data
               With Its Quality
             Peer Review of Data
               and Conclusions




General Guidelines:

A.   CERL Commitment to Comply to Laboratory Animal  Welfare

          The Corvallis Environmental  Research  Laboratory (CERL)  of the U.S.
     Environmental  Protection Agency (EPA) is committed to, adheres to, and
     will continue to adhere to the principles  and  recommendations set forth  in
     the NIH Guide for the Care and Use of Laboratory Animals and the princi-
     ples set forth by the "U.S. Government Principles  for the Utilization and
     Care of Vertebrate Animals Used in Testing,  Research, and Training."   CERL
     is also committed to complying with the Animal  Welfare Act passed by
     Congress in 1966 as P.L. 89-544 and amended  in 1970 and again in 1976
     (P.L. 94-279)  and all-other applicable Federal  statutes and  regulations.

B.   Committee Charge and Composition

         In order to promulgate this commitment to  comply, CERL has established
     an Institutional Animal Care and Use Committee (IACUC) to advise the
     Director on oversight of the Laboratory's  animal care program and to
     review research activities involving animals conducted at the Laboratory.
     This committee is composed of at least five  members, sufficiently quali-
     fied through experience and expertise to advise the Director on humane
     animal care and use.  Of these five members, at least one is a Doctor of
     Veterinary Medicine; at least one is a practicing  scientist  experienced  in
     research involving animals; at least one is  from a nonscientific area as
     his/her primary vocation; and at least one is  an individual  who is not
     otherwise affiliated with CERL and is not  a  member of the immediate family
     of a person who is affiliated with CERL.  The  names, position titles  and
     credentials of the IACUC members, the IACUC  chairperson and  the respons-
     ible institutional official (one who has the authority to sign commitments
     on behalf of CERL to EPA Headquarters) are on  file in the CERL Director's

C.   Function of the Committee

         (1) The IACUC is the principal advisory  body on humane care and use  of
     animals to the Laboratory and to researchers who use animals.  (2) The
     IACUC will resolve concerns within the Laboratory  involving  the care  and
     treatment of animals brought to the attention  of the committee.  (3)  The
     IACUC will recommend to the Laboratory Director termination  a project
     when, in its judgment, the conduct of the  research is such that compliance
     with the policy of humane treatment to animals is  jeopardized but only
     after suggested changes are made in writing  to the researcher, with a copy
     to the Branch Chief, and the researcher then fails to comply or satisfy
     the IACUC that altering the protocol will  compromise the research.  (4)
     The IACUC will  recommend to the Director of  CERL,  or his/her designated
     representative, changes and improvements regarding the Laboratory's animal
     care program or its animal facilities.

D.   Conduct of the Committee

         In the conduct of meeting the functions of the IACUC the committee

     1.   Review and approve the care and use of all  animals in  proposals
          submitted by a researcher for CERL approval  (i.e., fiscal  support or
          safety approval).  Proposals that reflect only a minor change  in  use
          (such as a different number of animals or length of test duration) in
          previously approved protocols may be approved by the chairperson  of
          the IACUC without full committee consideration.   The exception is
          when the IACUC chairperson is in direct conflict; i.e., serves as
          principal investigator on protocol under consideration.  In such
          cases, another IACUC member may be delegated to make a decision.
          Records of such actions are to be maintained in the committee  files.

     2.   Meet at least semiannually or when protocols are to be reviewed.   All
          decisions must be based on a majority vote of those members convened
          at a duly announced meeting.  The IACUC chairperson may poll members
          of the IACUC on project proposals when a delay in decision on
          approach or rejection would jeopardize the initiation  of a project.
          A majority vote must be recorded.

     3.   Conduct inspections of animal facilities and report the findings  of
          the inspections to the Branch Chief, the Director, and the researcher
          in charge of the animal facility inspected.   Such inspections  shall
          be made at least semiannually.

     4.   Prepare an annual report on CERL animal care and use to be submitted
          to the Director and/or EPA Headquarters.

     Review of all protocols to IACUC must include consideration of:

     1.   Species

     2.   Cage size adequacy

          (a)  animals per cage or per unit of size
          (b)  height

     3.   Chemical or biological administration

          (a)  radioactive compound
          (b)  potential carcinogen
          (c)  biological
          (d)  toxic substance

     4.   Anesthetic, analgesic, or tranquilizer to be used and  dosage.

     5.   Euthanasia methodology

          (a)  must comply with recommendations of the American Veterinary
               Medicine Association Panel  on Euthanasia (JAVMA, 1978,
               173:59-72); or

          (b)  approved by a veterinarian

     6.   Degree of pain inflicted by protocol  methodology

          (a)  restraint duration
          (b)  invasive procedures
          (c)  adequacy of diet


    The CERL Institutional Animal Care and Use  Committee has been selected, has
met for introductory discussions, and has  been  briefed on the operation and
responsibilities of the Committee.  The membership selected Dr. Dean Bauman as
the Chairman.

     The Committee has agreed to meet semiannually for general  reviews, and to
meet as needed to discuss and review new protocols.

     The Committee membership includes the following:

     Dean Bauman, D.V.M.:  Owner of Alpine Veterinarian Hospital
     5120 N.W. Highland Avenue
     Corvallis, Oregon  97330

     John Crawford, Ph.D.:  Associate Professor
     Department of Wildlife and Fisheries
     Oregon State University
     Corvallis, Oregon  97331

     Anne Fairbrother, D.V. M., Ph.D.:  Research Biologist and Staff Veterinarian
     Corvallis, Oregon  97333

     Eva Pendleton, D.V.M.:  Veterinarian  specializing in avian diseases
     Department of Poultry Science
     Oregon State University
     Corvallis, Oregon  97331

     Safa Shirazi, Ph.D.:  Systems Analyst in Environmental Sciences
     Corvallis, Oregon  97333

     Roger Zimmerman, Ph.D.:  Minister, First Christian Church
     4550 N.W. Queens
     Corvallis, Oregon  97330




     The development of knowledge necessary for the improvement  of  the health
and well-being both of man and of animals requires  recourse to HI vivo experi-
mentation with a wide variety of animal  species.  Methods such as mathematical
models, computer simulation, and in vitro biological  systems  should be used
wherever appropriate.  Whenever U.S.  Government agencies  develop requirements
for testing, research, or training procedures involving the use  of  vertebrate
animals, the following principles shall  be considered;  and whenever these
agencies actually perform or sponsor  such procedures,  the responsible institu-
tional official shall ensure that these principles  are  adhered to:

1.   The transportation, care, and use of animals shall be in accordance with
     the Animal Welfare Act (7 U.S.C. 2131 et.  seq.)  and  other applicable
     federal, state, and local laws and prescribed  policies.1

2.   Procedures involving animals should be designed and  performed  with due
     consideration of their relevance to human  or animal  health, the advance-
     ment of biological knowledge, or the good  of society.

3.   The animals selected for a procedure should be of  an appropriate species
     and quality, and the minimum number required to obtain scientifically
     valid results.

4.   Proper care of animals, including the avoidance or minimization of discom-
     fort, distress, or pain is a moral  imperative.  Lacking  evidence to the
     contrary, investigators should consider that procedures  that cause pain in
     human beings cause pain in other animals.

5.   Procedures with animals that may cause more than  momentary  or  slight  pain
     or distress should be performed  with appropriate  sedation,  analgesia, or
     anesthesia.  Surgical or other painful procedures  should not be performed
     on unanesthetized animals paralyzed by chemical  agents.

6.   Animals that would otherwise suffer severe or  chronic pain  or  distress
     that cannot be relieved should be painlessly killed  at the  end of the
     experiment or, if appropriate, during the  experiment.

7.   The living conditions of animals kept for  biomedical purposes  should
     contribute to their health and comfort.  Normally, the housing, care, and
     feeding of all animals used for  these purposes must  be supervised by  a
     properly qualified veterinarian.  In any case, veterinary care shall  be
     provided as indicated.

8.   Investigators and other personnel shall be appropriately qualified and
     experienced for conducting procedures on living animals. Adequate arrange-
     ments shall be made for their in-service training, including the proper
     and humane care and use of laboratory animals.

     If it is deemed necessary to waive one of the foregoing principles,  the
decision should be made, with due regard to the provisions of Principle 2,  by
an appropriate review board, such as an institutional  animal  research committee.
Such waivers should not be made where the primary purpose is teaching or

1 For guidance throughout these Principles, the reader is referred to Guide for
  the Care and Use of Laboratory Animals prepared by the Institute of Labora-
  tory Animal Resources, National Academy of Sciences.
Director, National  Institutes of Health