United States
             Environmental Protection
             Agency
           Office of Environmental Processes
           and Effects Research
           Washington DC 20460
EPA/600/9-86/004
January 1986 '/
             Research and Development
«»EPA
Report of the
Research Planning
Workshop on
Bioavailability of
Dioxins

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                                    EPA/600/9-86/004
                                    January 1986
REPORT OF THE RESEARCH PLANNING WORKSHOP
       ON BIOAVAILABILITY OF DIOXINS
               RALEIGH, N.C.
               SEPTEMBER 1984
   U.S. ENVIRONMENTAL PROTECTION AGENCY
    OFFICE OF RESEARCH AND DEVELOPMENT
           WASHINGTON, DC 20460

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                      NOTICE

This document has been reviewed in accordance with
U.S. Environmental Protection Agency policy and
approved for publication.  Mention of trade names
or commercial products does not constitute endorse-
ment or recommendation for use.
                       11

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                                  FOREWORD
      Research on the class of compounds called dioxins began several decades ago,
but the research activity increased substantially in the 1960s and  1970s, when the
complex  problem  of  dioxin  contamination  received  national  and  international
attention.   Although  considerable research has  been done  in this  field, there are
certain gaps in scientific knowledge, related to understanding the bioavailability of
dioxins,  that  need  to  be  identified  to  evaluate more  accurately  human  and
environmental risks associated with these chemicals.

      To accomplish this goal, the EPA Office of Research and Development sponsored
the Research Planning Workshop on the Bioavailability of  Dioxins,  September 9-12,
198*, that brought together scientists and  managers in various aspects of dioxin work
from  government agencies, academia, and industry.  About ninety researchers focused
their  attention during the four-day meeting on identifying the most obvious gaps in
knowledge and the consequent research needs.

      This report is the outcome of the workshop; it addresses the  current state of
knowledge on dioxins and defines the research  needs perceived by top scientific
experts in this field. Because of the range and complexity of this scientific area, the
report is divided into three main  parts to address  different  aspects of bioavailability:
environmental  processes  that   determine  bioavailability,  the  bioavailability  to
ecosystems, and the bioavailability to  humans.  This document is primarily intended
for use by the Agency to plan future research programs.  We  also hope this document
will be useful to other research organizations  in both  the government  and private
sectors.
                                       Erich Bretthauer
                                       Director
                                       Office of Environmental
                                         Processes and Effects
                                         Research
                                       m

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                                 CONTENTS


                                                                         Page

EXECUTIVE SUMMARY                                                       1

INTRODUCTION                                                              6

CHAPTER 1 - ENVIRONMENTAL PROCESSES IN BIOA VAIL ABILITY               8

   1.1  Introduction                                                          8

   1.2  Physical and Chemical Properties                                       9

   1.3  Transformation Processes                                             15

        1.3.1  Photochemical Processes                                       15
        1.3.2  Chemical Transformations                                     16
        1.3.3  Biological Processes                                           16
        1.3.*  Surrogate Parameters for Combustion                           17

   1.*  Transport Processes                                                  17

        1.4.1  Sorption/Desorption/Volatilization                              18
        1.4.2  Intermedia Transport                                          19
        1.4.3  Advection/Diffusion/Dispersion                                 19

   1.5  Modelling                                                           20

   1.6  Analytical Methodology for Analyses of TCDD in
        Environmental and Human Samples                                     21

CHAPTER 2 - BIOAVAILABILITY IN ECOSYSTEMS                              24

   2.1  Introduction                                                         24

   2.2  Exchanges of Dioxin Among Ecosystem Components                      26

        2.2.1   Conceptual Model                                             26
        2.2.2  Research Needs                                               26

   2.3  Bioavailability: Aquatic Ecosystems                                    26

        2.3.1   Introduction                                                  26
        2.3.2  Routes and Rates of Uptake, Metabolism, and Elimination          28
                                      iv

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                             CONTENTS (continued)
                                                                            Page
        2.3.3   Effects of Dioxin and Ecosystem Processes                       29
        2.3.4   Biological Decontamination Processes                           29
        2.3.5   Biological Effects                                              29
        2.3.6   Role of Food Chains in Human Exposure                         30
        2.3.7   Research Needs                                                31

   2.*  Bioavailability: Terrestrial Ecosystems                                 32

        2.4.1   Introduction                                                   32
        2.4.2   Degradation of TCDD in Soil                                    32
        2.4.3   Bioconcentration in Wildlife from Soil                           32
        2.4.4   Movement through Soil to Food Animals to Humans               33
        2.4.5   Movement through Soil to Plants                                34
        2.4.6   Research Needs                                                34

CHAPTER 3 - BIOAVAILABILITY TO HUMANS                                  37

   3.1  Introduction                                                           37

   3.2  Bioavailability of TCDD  to Humans from Environmental
        Matrices                                                              38

        3.2.1   General Properties of TCDD and Matrices                        38
        3.2.2   Research Needs                                                38

   3.3  In Vivo  Bioavailability                                                  38

        3.3.1   Mobilization and Redistribution                                  38
        3.3.2   Research Needs                                                39

   3.4  Host Factors Influencing Bioavailability                                 40

        3.4.1   Dietary Factors                                                40
        3.4.2   Genetic Differences                                            40
        3.4.3   Age                                                           40
        3.4.4   Concomitant Exposures                                         40
        3.4.5   Exposure History and Other Factors                              40
        3.4.6   Research Needs                                                41

   3.5  Interspecies Differences  Affecting Bioavailability                        41

        3.5.1   Dermal Route                                                  41
        3.5.1.1  Dermal Studies                                                 4'

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                             CONTENTS (continued)
                                                                            Page
         3.5.1.2 Research Needs                                                 42
         3.5.2   Oral Route (Ingestion)                                           42
         3.5.2.1 Oral Studies                                                    42
         3.5.2.2 Research Needs                                                 42
         3.5.3   Inhalation Route                                                43
         3.5.3.1 Inhalation Studies                                               43
         3.5.3.2 Research Needs                                                 44

   3.6   Pharmacokinetics and Structure-Activity Relationships                    44

         3.6.1   Pharmacokinetic Studies                                        44
         3.6.2   Research Needs                                                 44

   3.7   Epidemiology                                                          45

         3.7.1   Epidemiological Studies                                         45
         3.7.2   Research Needs                                                 46

   3.8   Need for Supply of TCDD                                               46

CHAPTER 4 - SUMMARY                                                       49

   4.1   Physical and Chemical Data                                             49

   4.2   Field Studies                                                           49

   4.3   Modelling                                                              49

   4.4   Summary                                                              50

APPENDIX -  Participants                                                       51
                                       vi

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                             ACKNOWLEDGMENTS


     The contributions of all  workshop  participants  are gratefully acknowledged,
especially Rizwanul Haque of EPA who served as chairperson of the workshop. The
success of the workshop and this report were possible because of the assistance of the
co-chairpersons:    Diane  Courtney,  Thomas  Duke,  and  Walter  Sanders,  U.S.
Environmental  Protection Agency;  Michael Gallo,  UMDNJ-Rutgers  Medical School;
Mark Harwell, Cornell University; and Captain Terry Stoddart, U.S. Air Force.

     The efforts of Dan Tisch,  Workshop Coordinator, and Linda Cooper, Technical
Editor, Northrop Services, Incorporated, in coordinating the workshop and in producing
this report are also acknowledged.  The final version of this report was integrated and
edited  by Christine C.  Harwell.   We also  thank Janice Wilson,  Word Processing
Specialist, Northrop Services, Incorporated, and Roberta Sardo and Carin Rundle  of
Cornell University for their efforts in producing this report.

                            PLANNING COMMITTEE

                          Rizwanul Haque - Chairman

     Donald Barnes                               James Falco
     Bruce Barrett                                Steve Jackson
     Judy Bellin                                  Harold Kibby
     Erich Bretthauer                             Barry Korb
     Michael Cook                                Robert Landers
     Phil Cook                                   Michael Mastracci
     Michael Dellarco                             Charles Nauman
     Paul desRosiers                              Ron Stanley
     Robert Dixon                                Frode Ulvedal
     Carl Enfield                                 James Upham
                                      vn

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                             EXECUTIVE SUMMARY
     The goal of the Research Planning Workshop on Unavailability of Dioxins was to
evaluate the ongoing research on the bioavailability of chlorinated dioxins and related
chemicals, to  identify research needs,  and to  develop a  focused research  plan.
Workshop participants were organized into three groups that addressed the topics:
environmental  processes  in  bioavailability,  bioavailability  to  ecosystems,  and
bioavailability to humans.

     Each group of participants at the workshop  addressed specific areas  regarding
bioavailability, within the broad range of  dioxin research,  and identified  areas  for
study in  their  final  summary reports.    However, the  definition and  concept of
bioavailability varied among the three groups. One group addressed dioxins in general,
while the other two groups focused discussions  on  2,3,7,8-tetrachlorodibenzo-g-dioxin
(2,3,7,8-TCDD).  Because the class  of dioxins represents a large number of chemicals
and  most attention to date has been  focused  on the isomer  2,3,7,8-TCDD, special
terminology is  used in this document to address these chemicals:   the term  TCDD
refers to 2,3,7,8-TCDD; the term dioxins refers to other isomers.

                    Environmental Processes In Bioavailability

     The group  dealing with the  environmental processes  in  the  bioavailability of
TCDD evaluated the  current state  of the art in analytical  methods, physical and
chemical properties, transport and  transformation  processes, and exposure modelling.
The group addressed the basic scientific understanding required  for valid estimation of
exposure, bioavailability, and risk.  The group identified major gaps in knowledge and
prioritized the  research needs in these areas. In ranking the research objectives, the
group considered  both short- and long-term needs.

Photochemical  Processes

     Available  information indicates  that  photolysis offers  the  most  promising
environmental  process for degrading TCDD.  Therefore, the highest research priority
was assigned to better characterization of  the rates and extent of direct and indirect
photolysis in air, on surfaces, and in water.

Physical and Chemical Properties

     The second research priority  concerns the expansion of the data base covering
physical and chemical properties  of TCDD and  related compounds.   The  use of
structure-activity relationships, based on thermodynamic laws, to predict physical and
chemical  properties  offers a  cost-effective  alternative to  numerous laboratory
determinations.

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Sorption/Desorption/Volatilization

      To understand better  the dynamics of  TCDD movement in all environmental
media, extensive studies of  sorption/desorption phenomena are needed.  Specifically,
the effects of organic  content of soils and  sediments on the sorption/desorption of
TCDD from  the saturated  and  unsaturated  zones require further study, as do the
effects of particulates on TCDD in the atmosphere.

Chemical Transformations

      The  chemical transformations  of TCDD  have  not been  characterized.   A
complete definition of  the  transformation processes will contribute  greatly to the
understanding of transport as well as provide indications of specific chemical reactions
that  may  be  employed to degrade  TCDD.   The  ability  of TCDD  to  undergo
oxidation/reduction,  acid/base  hydrolysis,   nucleophilic  displacement,  and  metal
chelation reactions needs to  be determined.

Biological Processes

      The use of biodegradation as a  cost-effective procedure for TCDD  degradation
must  be  ranked as a long-term, high-priority research need.  Because the genes for
TCDD metabolism have been demonstrated in higher organisms, the employment of
recombinant   DNA technology to  construct  a  microorganism  capable  of TCDD
degradation may greatly benefit future cleanup operations and reduce the risk to the
environment and human  populations.

Intermedia Transport

      Although the rates of certain intermedia transfers can be predicted,  appropriate
measurement  techniques are not  available to validate  the predictions.   The bulk
transport and intermedia transfer of TCDD  require further  characterization.   The
movement  of TCDD-contaminated particles  appears to  be  a critical link in the
bioavailability of TCDD.

Modelling

      Currently available mathematical exposure models are applicable to predict the
exposure concentrations of TCDD in the various environmental  media,  if and when the
various equilibrium and  rate coefficients have been determined.  However, research
will be  required to apply  and test the various  media  and  multimedia models  to
determine their degree of applicability, precision, and accuracy.

Surrogate Parameters for Combustion

      The  production of  dioxins  and furans from  municipal  incinerators is  well
documented.  Identification and measurement of these chemical species are costly. To
control effectively the combustion process so  that the release of these compounds is
eliminated, new surrogate parameters for process control are needed.

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Need for Sufficient Supplies of TCOO for Research

     An adequate supply of TCDD and laboratory standards for all possible isomers of
dioxins  and furans is not readily  available.  These materials are required for  quality
assurance and  quality control and  to  facilitate investigations of  dioxin and fur an
distribution in the environment. An appropriate toxicological evaluation of dioxins and
furans cannot be conducted without the appropriate laboratory standards.

                           Bioavailability In Ecosystems

     To gain a better understanding of the pathways to humans  and effects  on the
ecosystem,  the fate and transport of dioxin in the environment must be determined.
This determination includes addressing areas such as the bioavailability of dioxin to
biota, and how  organisms influence the transport of dioxins within and across systems.
Also of  interest are rates of exchange, such as the rate of dioxin uptake by plants from
soil, and the ultimate partitioning  of the chemical.

     To clarify  such interactions, results  of  group discussion  are  presented  in  a
section   on  a  conceptual framework  for  exchanges  of dioxin  among ecosystem
components, followed by sections on aquatic and terrestrial ecosystems with respect
to fate, transport, effects on biota (species and processes), and pathways to humans.

     In  these sections,  three sets  of  topics were considered:   1) identification of
ecosystem  processes that a.) are involved in routes, rates, and  reservoirs of dioxins in
aquatic and terrestrial ecosystems; b.) are  particularly susceptible  to effects of dioxin
contamination;  or c.)  are involved  in biological  decontamination  processes;   2)
identification of  particular species and communities that are impacted or potentially
impacted by dioxins; and   3) identification of the role of food  chains  and webs in
human exposure and risk.

Research Needs

     Data on the impact of dioxins at the ecosystem level are essentially nonexistent,
and  relatively  few data are available  describing  the effects  of these  chemicals on
single species.  Therefore, many research needs were identified by group participants;
the following were considered the  highest priority:

  •  Develop  the capability to  predict  dioxin  levels in tissues (particularly in
     organisms that  constitute human food chains) as a  function  of environmental
     conditions;  develop toxicity data for  understanding the  mechanisms of toxicity
     and the factors responsible for differences in sensitivities  among species.

  •  Measure the concentration  of dioxins  over time  in organisms as a function of
     dose in food, water, and other sources  for model development.  Use microcosms
     for model verification.

  •  Conduct a  full-scale ecological study  at  a highly contaminated site.    Include
     field  studies of  fate, chronic effects, and ecological processes,  with supporting
     laboratory studies.

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  •  Evaluate   the   chemical  and   biological   characteristics   of  residue   from
     experimental incineration projects and incorporate results in risk assessment.

     In addition to these research needs, there is a need to improve risk assessment
capability  and  to  evaluate  the  uncertainties  resulting  from  conflicting   data,
unexpected indirect effects, and laboratory-to-field extrapolations.

                            Bioavailability To Humans

     Risks may be inaccurately estimated in the absence of knowledge about factors
determining bioavailability, even when exposure is relatively well defined.  Clearly,
matrix and route effects are likely to be significant.  However, human responses and
risk are also influenced by exposure and by differences in the sensitivity of target sites
of action.  To  consider bioavailability adequately, exposure and toxic  response must
also be examined.

     The following research needs were identified to evaluate the bioavailability of
TCDD relative  to human health.

Matrices

     The bioavailability  of TCDD from matrices of soil, fly ash,  and respirable
particles should be determined using the same species and same toxicologic end points.
A range of concentrations should be utilized, because the bioavailability of TCDD may
differ at differing concentrations levels.

Host Factors; Deposition and Mobilization of TCDD

     Because  of the lack of knowledge of the  critical target organ(s) in  humans,
studies are needed to determine the  appropriate animal species to use as models for
studying host  factors,  tissue  distribution,  and  mobilization  from   body  stores.
Additionally, the  critical end  points  and other biochemical  markers  need  to  be
determined for  both human and other animal models.

     One of the sensitive end points in animals and possibly humans  is the immune
system.  The data that are available indicate further studies are needed of the effects
of TCDD on the immune system.

     The body  burden of TCDD in humans needs to be determined using adipose tissue
as the most important depot. Studies determining the residue of TCDD in  other organs
might  indicate possible  target  organs,  as well  as the mobilization,  redistribution,
metabolic pathways, and secretion or excretion patterns in humans.

Pharmacokinetics and Structure-Activity Relationships

     Because humans are often exposed to mixtures of compounds  that would include
TCDD  and similar  chemicals, studies are needed to delineate the interactive effects c f
dioxin  and furan isomers with  TCDD, and to determine the additive,  synergistic, or
antagonistic effects, as well as the pharmacodynamics of the mixtures and receptor
level modulations.

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Extrapolation of Animal Data to Humans

     An animal model that  best  indicates TCOD  toxicity in humans  is still being
developed according to the criteria more fully discussed in Chapter 3.  Because there
are so  many manifestations of TCDD toxicity, it may be necessary to have more than
one model, depending on the end point.

Epidemiological Studies

     Methods need to  be  developed to identify persons who have  been exposed to
TCDD  and related compounds as a basis for  epidemiological  studies.  Additional
studies in humans  should be done with  cohorts not exposed to TCDD to establish the
baseline for the end points of toxicity. Rigorous epidemiological studies with sound
methods and  proper execution  are needed to  determine  the effects  of  TCDD in
humans.   Until there are better epidemiological studies, the determination of TCDD
toxicity will not be known with any assurance, and the extrapolation of animal data to
humans cannot be done  reliably.

Assay of TCDD

     To perform  many  of the  suggested studies,  there is a need to develop  and
validate assays of TCDD that are rapid and economical, either in vivo or in vitro, and
that can be used to determine the concentration of TCDD in various organs.

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                                INTRODUCTION

     The class  of  chemicals polychlorinated dibenzo  dioxins, commonly  known  as
dioxins, has attracted great attention and raised controversies during recent years.  In
the United States, issues about dioxins surfaced  during the  1960s,  when  2,3,7,8-
tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) was  found to be  a contaminant in the
commonly  used herbicide  2,4,5-T (2,^,5-trichlorophenoxy-acetic  acid).   The  high
toxicity  and  persistence of 2,3,7,8-TCDD in the environment represent the  primary
characteristics of dioxins that  pose risks  to  human health  and  the environment.
Because  the class dioxins represents a large number of chemicals and most attention
to date has been focused on the isomer 2,3,7,8-TCDD, special  terminology is used in
this document to address these  chemicals:  the term TCDD refers to  2,3,7,8-TCDD;
the term dioxins refers to other isomers.

     Since the 1960s, several incidents  have focused attention on  the  contamination
problem: the human and environmental exposure to dioxins as a result  of a chemical
plant accident in Seveso, Italy; the identification of dioxins at several hazardous waste
sites in  the  states of  Missouri,  New Jersey,  New York,  and Arkansas;  and the
occurrence of dioxins in fish samples in the states of Michigan and Wisconsin.  Dioxins
are also  associated  with combustion processes and are found in municipal incinerator
fly ash.

     In spite of the release of dioxins to the environment and concomitant potential
exposure of humans, the pathways and persistence have not been fully investigated for
many   environments.    Additional  study is required on dioxin  accumulation  and
partitioning in living systems, the toxicity associated with dioxins, the evaluation  of
the human and  environmental risks, and the  development  of control technologies
necessary to  minimize such risks.  This information is crucial  to making appropriate
regulatory  decisions  about dioxins  under  the  Federal  Insecticide, Fungicide,  and
Rodenticide Act (FIFRA), the Toxic Substances  Control  Act  (TSCA),  the  Resource
Conservation  and  Recovery  Act  (RCRA), and  the Comprehensive  Environmental
Response Compensation and Liability Act (CERCLA or Super!und).

     Central to the complex issues of exposure and risk assessment is the evaluation
of the  bioavailability of dioxins.  The term bioavailability has not been clearly defined,
and the subject remains  poorly understood.  It  involves the understanding of factors
related in  the uptake, release, or bioaccumulation of dioxins by living organisms.
Recent findings  at the National Institute of Environmental Health Sciences indicate
that TCDD was  bioavailable to laboratory animals fed  with contaminated soils from
Missouri.  But a similar experiment done with New Jersey soil indicated that dioxin
was not  detected in the  bodies of  laboratory animals.   To understand the  complex
issues of  bioavailability, the environmental processes that can influence bioavailability
to ecosystems and humans must be characterized.  Identification and quantification  of
TCDD  and  other dioxins  in environmental and  biological matrices also  require major

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attention.  Most research data deal with TCDD; the toxic effects and environmental
risks associated with other dioxins also need to be evaluated.

      Dioxin research currently funded by the U.S. Environmental Protection Agency
(EPA) addresses four main areas: 1) development of measurement methods and quality
assurance  procedures  for identifying and quantifying dioxins;   2) development  and
evaluation  of control technologies for containment and destruction of dioxins; 3) study
of the fate of  dioxins in soils and investigation of the uptake of dioxins by plants and
animals; and  4) assessment of health and environmental risk associated with dioxins.

      Because understanding the bioavailability of dioxins is essential to understanding
toxicity and risk, a workshop was held to obtain input in developing a focused research
plan.  Scientists attending represented a broad range of expertise from research groups
in academia, industry, and government organizations. The goal of this workshop  was
to  evaluate the ongoing research on the bioavailability of chlorinated  dioxins  and
related chemicals, identify research  needs, and  develop a focused  research plan.
Workshop participants were organized into three groups that addressed the topics: 1)
environmental  processes in  bioavailability;   2) bioavailability to ecosystems; and  3)
bioavailability to humans.

      The Environmental Processes in Bioavailability Group focused on defining various
environmental  processes  controlling  the  bioavailability  of TCDD  in  the biosphere.
Transformation processes and bioavailability assessments were also discussed in  this
group. The Bioavailability to Ecosystems Group evaluated the factors relevant to the
bioavailability  of dioxins in aquatic  and terrestrial ecosystems, and the potential
impact  of  these chemicals on ecosystems.   Ecosystem processes were  identified,
particular  species and  communities  that are potentially  impacted by dioxins were
addressed,  and the role of food chains and the food web in human exposure and risks
were  discussed.  The Bioavailability to Humans Group evaluated factors such as the
bioavailability  of TCDD from  environmental matrices, the host  factors  affecting
bioavailability,  in vivo bioavailability,  routes of  exposure,  toxic  human  effects,
inter species differences, and extrapolation from other animals to humans.

      This  report will  be used by  EPA in its research  planning.  We hope that  this
report will be beneficial to other research organizations in planning their research.

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                                  CHAPTER 1

               ENVIRONMENTAL PROCESSES IN BIOAVAILABILITY

          Co-chairpersons:  Walter M. Sanders III and Capt. Terry Stoddart


                                1.1 Introduction

      The presence of TCDD in the environment  poses great concern because of its
known toxicity and  persistence.    Bioavailability of  TCDD and  other chlorinated
aromatic  compounds in the environment depends  on many factors  that  control their
concentration in the biosphere.   Important factors include physical  and chemical
properties,  transport  and  transformation  processes,  and  characteristics  of  the
environmental media.

      One workshop group of scientists and engineers addressed the many complex
physical and  chemical issues related to the  bioavailability of TCDD.  The group:  1)
evaluated   available   analytical   methods,   possible   transport   pathways,  and
environmental  transformation  processes;   2) reviewed  the current  state  of  the
understanding of bioavailability of  dioxins; 3) identified significant research needs; and
*) prioritized the efforts  required to bring  the state of the  art  up to an acceptable
scientific level  to understand human and environmental exposure and risk assessment.

      Also considered were the needs for significant supplies of TCDD for research and
standards/reference  samples  for  quality  assurance  and quality  control,  the
applicability  of available  exposure  modelling techniques specifically for TCDD, and
the need for  surrogate parameters  for combustion  processes control. The consensus of
the group was  that the fate of dioxin isomers and related chemicals, such as the
polychlorinated dibenzof urans and xanthenes, should be considered along with TCDD as
human and environmental toxicology dictates.

      The  seven highest  priority research needs identified by the group are discussed in
order of  importance, with a statement concerning the  current state of the art and
research   objectives.    Three  related  items,  analytical  methods,  modelling,  and
surrogate  parameters for combustion, were given equal priority, and  are also identified
below.

      Ranking                           Need

        1                     Photochemical Processes
        2                    Physical and  Chemical Properties
        3                    Sorption/Desorption/Volatilization
        *                    Chemical Transformations
                                       8

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     Ranking                           Need

         5                     Biological Processes (Longer-term)
         6                     Intermedia Transport
         7                     Advection/Diffusion/Dispersion Activities
         o                     Analytical Methodology for Analysis of TCDD in
                                 Environmental and Human Samples Modeling
         o                     Modelling
         o                     Surrogate Parameters for Combustion

     These  prioritized  research  needs have  been grouped  for  discussion into five
categories:   Physical and Chemical Properties, Transformation Processes, Transport
Processes, Modelling, and Analytical Methodology for Analysis.

     In some cases, the  research suggested  by this working group related only to
TCDD, such as the measurement or validation of the physical and chemical properties
of TCDD.  In other  cases, the recommended  research extends the  current scientific
state of the art for   characterizing environmental  transport  and  transformation
processes and will be applicable to other hydrophobic organic chemicals.  This would
include research efforts to characterize, identify, and measure the soil characteristics
(e.g., organic and moisture content, particle distribution, and  temperature) that govern
the rates and extent of the  various transport  and degradation processes important to
TCDD and all other hydrophobic organic chemicals.

     Results from the research outlined in this section will have major impacts on the
understanding and estimation of  the bioavailability of  TCDD to  both humans  and
ecosystem components.  For example, if volatilization from sorbed surfaces at night is
a significant transport  pathway, inhalation or  dermal contact with vapor-phase TCDD
will be an exposure route that must be considered. Results from this  research will also
apply directly to the modification of regulatory criteria and  standards by providing
more accurate identifications of exposure pathways and rates.  Likewise, results may
impact the treatment and control of  TCDD-contaminated areas if direct vapor-phase
photolysis or biodegradation mediated by genetically engineered microorganisms could
be incorporated into  significant in-place treatment processes.

                       1.2 Physical And Chemical Properties

     The known physical and chemical properties for TCDD  are summarized in  Table
1.1, which includes literature values for measured and estimated properties and values
measured by Schroy and associates.   Values  not available from laboratory work or
literature were estimated.  Comparable data for other dioxins, furans, and other very
low volatility chemicals are not available.

Research Needs

     Reasonable estimates  of physical  and chemical properties of TCDD isomers,
other dioxins, and f urans in general are necessary to permit  rational analysis of their
behaviorwithin the biosphere.  The structure-activity relationship approach, based on
thermodynamic consideration, represents a cost-effective alternative for this purpose

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                                                     Table 1.1

                                         PHYSICAL PROPERTY DATA SHEET*
CHEMICAL NAME
CHEMICAL FORMULA
MOLECULAR WEIGHT
2,3,7,8-Tetrachlorodibenzo-
p-dioxin
(Synonym 2,3,7,8-TCOD or Dioxin)
CAS//-  17*6-01 -6
   321.97*
(IUPAC)
                 SPECIFIC HEAT,a/(°K-gmol)
                 gas  @25°C
HEAT OF VAPORIZATION
 @NBP, kj/gmol
 @NBP, BTU/lb

 @MP, kj/gmol
 @MP, cal/g
 @MP, BTU/lb
                                                                                              250.92*
 71.81*
 95.95*

 85.00*
 63.10*
113.57*
                                                 (SHA)
(RPS)
(RPS)

(MON)
(MON)
(MON)
CRITICAL CONSTANTS
 P = Pressure, Pa
 T = Temperature, K
 Z = Compressibility
 V = Volume, cm3

DENSITY, g/ml
 Solid @ 25°C
 Solid/Liquid @ MP
 Liquid @ NBP

VAPOR PRESSURE
 Pascals® 30.1°C
 Pascals @ 5*.6°C
 Pascals @ 62.QC
 Pascals @ 71 .°C
  2372.829*
   93*.5   *
     0.233*
   763     *
     1.827*
     1.720*
     1.021*
     *.68 E-7
     1.8* E-5
     5.03 E-5
     1.61 E-*
(RPS)
(RPS)
(RPS)
(RPS)
(BRM)
(RPS)
(RPS)
(MON)
(MON)
(MON)
(MON)
HEAT OF FUSION
 @MP, kJ/gmol                       38.91
 OMP, cal/g                         28.88
 (3MP, BTU/lb                       51.99

HEAT OF SUBLIMATION
 @MP, k/gmol                       123.91
 (§MP, cal/g                         91.98
 @MP, BTU/lb                      165.56

HEAT OF FORMATION
 gas   @25°C, k/gmol               -205.*3*

FREE ENERGY OF FORMATION
 gas   @25°C, kJ/gmol               -195.18*
               (BRM)
               (BRM)
               (BRM)
               (MON)
               (MON)
               (MON)
               (SFT)


               (SHA)

-------
                                                  Table 1.1 (cont.)

                                          PHYSICAL PROPERTY DATA SHEET
ANTOINE CONSTANTS
 Units   log e, Pa & K
Temp. Range
  A
  B
  C
T = 10 to 420°C

  10 to 305°C
  34.57083
  14903.438
  0.00
  (MOM)
NORMAL BOILING POINT (NBP) @ 1 atm
 Degrees C                   421.4  *
 Degrees K                   684.52*
 Degrees F                   790.5  *
 Degrees R                   1250.1**
FREEZING POINT
                             305.0
303 to 420=>C
25.10*35*
9430.391*
0.00*
(MON)
                     (MON)
                     (MON)
                     (MON)
                     (MON)
                     (BRM)
SOLUBILITY IN (@ I atm)
                  T = °C milligrams/liter
Water
Agent Orange
o-Dichloro-
benzene
Chlorobenzene
Benzene
Chloroform
n-Octanol
22
25

25
25
25
25
25
0.00000791
5&0

1400
720
570
370
48
                                            (ADB)
                                            I-VA)

                                            (ETD)
                                            (ETD)
                                            (ETD)
                                            (ETD)
                                            (ETD)
HEAT OF COMBUSTION
gas  @25°C, kj/gmol

ENTROPHY @ 101.325kPa
gas  (9250C, k3(OK-gmol)
-5000.3*
                                                                                              478.06*
                                                            SCHMIDT NUMBER (vapor in air @ dilute cone.)
                                                             Dimensionless 
-------
                                                         Table 1.1 (cont.)

                                                PHYSICAL PROPERTY DATA SHEET
ro
   Methanol
   Acetone
25
25
 10
110
(ETD)
(ETD)
ACTIVITY COEFFICIENTS AT INFINITE DILUTION IN

Water
Agent Orange
o-Dichloro-
benzene
Chlorobenzene
Benzene
Chloroform
n-Octanol
Methanol
Acetone
T =
22
25

25
25
25
25
25
25
25
°C GAMMA
2.26 E12
2368

20*3
439*
6356
10854
4270
796000
39864

(CALC)
(CALC)

(CALC)
(CALC)
(CALC)
(CALC)
(CALC)
(CALC)
(CALC)
   HENRY'S LAW CONSTANT FOR AIR/WATER SYSTEM
   H, atm m3/gmol           4.88        E-5*      (CALC)
Benzene
Chloroform
n-Octanol
Methanol
Acetone
25
25
25
25
25
1.3
2.25
8.8
1.6
8.25
E-8
E-8
E-8
E-6
E-8
(CALC)
(CALC)
(CALC)
(CALC)
(CALC)
                                                                    ENVIRONMENTAL FACTORS
                                                                    Oxygen Demand
                                                                    ThOD, Ib O2/lb chemical             1.193            (CALC)

                                                                    IMMEDIATELY DANGEROUS TO LIFE AND/OR HEALTH
                                              ppm                                 0.001*

                                             SAFETY
                                              Acute Toxicity
                                              oral (guinea pigs) LD LO, mg/kg       0.0006
                                                                                                                        (CDC)
                                                                                                                        (CAD)
     Prepared by J.M. Schroy (1984). Physical properties without asterisks (*) are reported as measured values, but those with asterisks have
     been reported as estimated by the author referenced or by a protocol in the referenced document.  Those physical properties marked
     with an asterisk and referenced as calculated were developed from other data on this sheet by Schroy. Values from laboratory work or
     literature are referenced as in the Table 1.1 References.  Values not available from laboratory work or the literature were estimated
     using methods given by Reid et al. (1977), and are identified as RPS.

-------
                            References for Table 1.1
 ADA
 BRM



 CAD


 CALC



 CDC


 ETD



 IUPAC


KEN



MON
RPS


SHA


SFT
 Adams,  W.3., and  Elaine, K.M..   "A Water  Solubility of  2,3,7,8-TCDD",
 Monsanto Co., St. Louis, Missouri, Dioxin 85 - 5th International Symposium
 on Chlorinated Dioxins and Related Compounds, Byreuth, FGR, September
 16-19, 1985.

 Boer,  P.P.,  van  Remoortere,  P.P., and Muelder,  W.W.   (1972).   The
 preparation and structure of 2,3,7,8-tetrachloro-p-dioxin and 2,7-Dichloro-
 p-dioxin. Journal of the American Chemical Society, 94(3).
 Casarett, L.J., and  Doull, J.   (1980).  Toxicology;
 Poisons. 2nd Ed. Macmillan, New York.
The Basic Science of
Calculation based on other  physical properties.  Activity coefficient based
on water solubility.  Theoretical oxygen demand based on molecular weight.
Partition coefficient for air/water based on activity coefficient.

Centers  for Disease  Control,  U.S. Department of  Health and Human
Services, personal communication. (1982)

Esposito,  M.P.,  Teirnan, T.O., and Dryden, F.E.   (1980).  Dioxins.  EPA-
600/2-80-197.  U.S.  Environmental  Protection Agency, Office of Research
and Development, Washington, D.C.

International Union of Pure and Applied Chemistry. (1979). Atomic weights
of the elements 1977. In Pure and Applied Chemistry.  51:405-533.

Kenaga, E.E.  (1980). Correlation of bioconcentration factors of chemicals
in aquatic  and  terrestrial  organisms with  their physical and  chemical
properties.  Environmental Science and Technology, 14(5),  553-556.

Cheng, S.C., Hileman, F.E., and Schroy, 5.M.  (1984).   Measurements of
vapor pressure at four temperature levels, and development of the heat of
sublimation from  the  correlation of  the vapor  pressure data using the
Clausius-Clapeyron equation; estimates of the heat of vaporization were
made using the  measured  heat  of  sublimation  and the  heat  of fusion.
Monsanto Company, Physical Property Research, Dayton,  OH and St. Louis,
MO.

Reid, R.C., Prausnitz, 3.M.  and Sherwood, T.K.  (1977).  The Properties of
Gases and Liquids. 3rd Ed. McGraw-Hill, New York.

Shaub, W.M.   (1982).    Estimated thermodynamic  functions  for some
chlorinated benzenes, phenols, and dioxins. Thermochemica Acta, 58, 11-44.

Seaton, W.H.,  Freedman, E., and Treweek,  D.N.  (1974).  CHETAH -  The
ASTM Chemical Thermodynamic and Energy Release Evaluation Program.
ASTM DS 51. American Society for Testing and Materials.
                                      13

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VA      3RB Associates.  (1980).  Review of Literature on Herbicides, Including
         Phenoxy  Herbicides and Associated  Dioxins.   Volume  I.   Analysis of
         Literature.  Veterans Administration, Department of Medicine, Washington,
         D.C.

-------
because the agents of concern  are  primarily rigid-ring systems.   This approach will
permit  the  estimation  of  Henry's  Law  Constant  and  distributive  tendencies.
Estimation of partitioning behavior across organic liquid/water, air/water, and organic
liquid/air  interfaces can then  be made.  This will  aid estimates of  mass transfer
processes   such  as  sorption/desorption,   volatilization,   diffusion/dispersion,  and
advection.  These data are critical to understanding and ultimately controlling the
human and environmental impact of these low volatility materials.

                          1.3  Transformation Processes

     Important   transformation  processes   considered  by  the  group  included
photochemical  processes,  chemical   transformations,  biological  processes,  and
surrogate  parameters for combustion.

1.3.1  Photochemical Processes

     As the dominant transformation process for TCDD in the environment, photolysis
offers  the  most  promise for  degrading dioxins in  air,  on soil,  and  in  water.
Experiments  by workshop  participants and  others have demonstrated that sunlight
photolysis  of  TCDD in  organic  media, water, and on  some surfaces may be rapid.
Other studies  have indicated that the transport of TCDD occurs slowly but measurably
to the soil surface, where photolysis can occur either on the soil surface or in the
vapor phase above it. TCDD can be introduced into the atmosphere from incinerators
and other  incomplete combustion processes and by volatilization from water. Sorption
onto airborne particulates and subsequent photolysis are also  possible.   All of these
degradation processes serve to reduce TCDD bioavailability in the environment.

     Demonstrated  degradation  pathways  for  photolysis  of TCDD involve  both
reduction  and ring cleavage, but other pathways  are possible, especially in  water and
air.  Rate constants for sunlight photolysis in water and organic media are reasonably
well known.   However,  little or no  data exist for photolysis of TCDD in  the vapor
phase or for TCDD sorbed onto airborne particulates (e.g., fly ash, dust), or onto soil
particles.

Research Needs

     The  primary research needs in this area include:

     •  measurements  of the rate constants, quantum  yields, and pathways of vapor-
        phase photolysis of TCDD in the solar range of light;

     •  similar  measurements  of  TCDD sorbed onto airborne particulates, soil
        surfaces, and environmental adsorbents; and

     •  determination of major pathways for photolysis in solution.

     Secondary research  needs include similar  measurements on other chlorinated
dioxins and related compounds.
                                       15

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1.3.2  Chemical Transformations

      Little work has been done on characterizing the chemical degradation reactions
of dioxins or f urans.  These chemicals are somewhat refractory to certain reactions, as
is the case with other highly halogenated aromatics.  Few of these  reactions have
been studied adequately, except the reduction involving alkaline glycol.

      Highly  charged metal  ions  are  capable of splitting aromatic  ethers  when  a
driving   force   is   available  through   chelate  ring  formation.     Thus,   1,2-
dimethoxyanthraquione is  selectively demethylated at the 1-position by Sn(lV) chloride
to form the corresponding tin chelate.  Such reactions suggest the possibility that tri-
and tetravalent  metal ions can assist in splitting the ether bridges of  dioxins to form
the very stable catecholate chelates of these metal ions.  In  addition,  the loss of one
chlorine atom from TCDD results in a marked reduction of toxicity.  Cleavage of the
oxygen linkages also produces a much less toxic product.

Research Needs

      A study of chemical reactions of the dioxins, f urans, and related compounds will
provide information for exposure estimates and other methods of altering the  toxicity
of these compounds.  The types of reactions that should be studied include:

      •   dechlorination by alkaline-glycol treatment;

      •   oxidation and reduction by various reagents and catalysts, including metal
         oxides;

      •   ether cleavage with  various reagents, including highly charged metal ions;

      •   metal complexation  and coordination; and

      •   other types of reactions that future research may indicate.

1.3.3  Biological Processes

      Biological  methods  for TCDD  degradation may offer  significant economic
advantages over other technologies.  Despite the evident persistence of TCDD  in soil,
there are indications  that it  can be microbially  detoxified. Because of its structure,
the initial step in the microbial transformation of  TCDD requires either hydroxylation
by  an  oxygenase or a  dechlorination  reaction.   Some  microorganisms  with  a
monooxygenase system have been shown to degrade TCDD by incidental metablism to
a  hydroxylated  product.   The rate  of  this transformation can  be increased  by
facilitating cellular uptake and by using a suitable substrate  to induce the necessary
enzymes.  The hydroxylated  products  formed are expected to be less toxic  because
they can be biologically conjugated and eliminated; they are also more water  soluble
and dispersible.  Fungi and yeasts  are known to  have a broad range of  monooxygenase
enzymes,  including  some that attack  polycyclic  aromatic  hydrocarbons  (PAHs).
Mammalian monooxygenases  have  been implicated in hydroxylation and dechlorination
of TCDD, and  a mammalian monooxygenase  gene  has been transferred  to  and
                                       16

-------
expressed in yeast.  The possibility of reductive dehalogenation by microorganisms in
anaerobic environments also exists.

Research Needs

     Microbiai degradation of TCOD in contaminated soil or sediment requires the
establishment  of microorganisms  that:   can  survive  and thrive in the necessary
environments; uptake TCOO in low concentrations; and contain an oxygenase system
that can dechlorinate or hydroxylate  TCDD.  Research approaches to obtaining such
strains are to:

     •   screen and  genetically  modify  bacteria, yeasts, and fungi that express
         monooxygenases;

     •   construct bacterial systems through a continuous culture selection procedure;
         and

     •   bioengineer  yeasts  or  fungi  for   expression  of   suitable  mammalian
         monooxygenase activities.

     Parallel  studies are required to facilitate or genetically enhance the uptake of
TCDD.   This  may  be accomplished by adding  solvents or surfactants, or by  using
surfactant-producing species.

1.3.4 Surrogate Parameters for Combustion

     There exist today a  large number and variety  of  facilities utilizing numerous
feedstocks and  combustion  processes  that   may   emit   TCDD  in  measurable
concentrations.   Present  methods  of measuring  TCDD  in stack  emissions  from
combustion sources are extremely complex and prohibitively expensive.  In addition,
the length of time  for stack sample analysis prevents immediate feedback to the
system operator for controlling TCDD emissions.  Present research in this area has
been confined  to  drawing limited correlations between TCDD emission levels and
regularly monitored combustion parameters.

Research Needs

     A research effort is  needed  to determine if inexpensive measurements of real-
time combustion processes can be  correlated with TCDD emissions and can  be  useful
in controlling TCDD emissions from combustion  sources.

                             1.4  Transport Processes

     The  important  transport   processes  considered   by  the  group  included
sorption/desorption/volatilization,   intermedia  transport,  and   advection/diffusion/
dispersion.
                                       17

-------
1.4.1  Sorption/Desorption/ Volatilization

      Considerable data are available to characterize the adsorption and partitioning
of organic chemicals, particularly pesticides, aromatic hydrocarbons,  and PCBs, on
participate matter derived from  soils and sediments.  This has led to a number of
correlations for estimating adsorption by soils for chemicals whose adsorption has not
been  measured. Organic matter content is considered to be the factor controlling the
extent of adsorption  and  has led to the use of the Koc (octanol/water coefficient)
concept for estimating adsorption.

      Soil/water partition  coefficients (Kj) of 10^ ml*g~l have  been  calculated for
TCDD,  based  on  measured values  for  water solubility and  estimated  values for
octanol/water partition coefficients.  Apparent TCDD  partition  coefficients ranging
from  10* to 10° have  been estimated, utilizing leachate data from a few contaminated
soils from Missouri and New Jersey.

      No information  is available on partitioning behavior with respect to TCDD on
atmospheric participates.   However, it has been postulated that the binding of TCDD
to fly ash during combustion is irreversible.

      Considerable information has appeared in the literature on the  application of
soil/water and air/water partition coefficients (Henry's Law Constant) to the transport
of  organics  to the  surface  of   a  soil  column,  where  they  will  be  subject to
volatilization.   Currently, the estimated partition coefficients  for TCDD  must be
relied upon for determining the volatilization potential for dioxins.

Research Needs

      The primary research needs in this area include:

      •   determining  rates  and extent of adsorption/desorption in soil and sediment
         systems as  a function  of organic  matter  and mineral content, either to
         validate or to reject the Koc concept for predicting partitioning;

      •   identifying  the mechanisms  influencing TCDD  partitioning  onto  mineral
         surfaces that are characteristic of soils having low organic matter content;

      •   determining  adsorption/desorption on atmospheric particulates as a function
         of organic matter, mineral, and water content;

      •   characterizing the vapor-phase desorption  isotherms  as a function  of
         participate (soil, sediment, or air) composition and water  content; and

      •   identifying the factors that would alter  the predicted compound behavior in
         soil/sediment systems.   Factors that  should  be investigated  include:   the
         presence of  strong acids  and bases or high levels of dissolved organic matter
         in  interstitial waters;  the presence  of co-solvents;  the  movement  cf
         microparticulates, organic colloids, and colloidal clays; and the influence of
         organic micelles (or emulsions).
                                       18

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1.4.2 Intermedia Transport

      Theoretical kinetic analyses of TCDD interphase transport have received limited
attention.    Low volatility  substances  such  as  TCDD  have  enormous  activity
coefficients in water. In volatilizing from  water,  TCDD probably forms a liquid film
(two-dimensional gas) on the water surface.  Thus, its transport  to  the gas phase
probably depends directly on its  thermodynamic activity.   Transport  between other
phases  probably  operates in  more  complex  ways.  No detailed analyses  of these
transport systems are available.  The lack of essential validated physical and chemical
data currently makes analyses of intermedia transport very difficult.

Research Needs

      The mechanism for intermedia transport must be characterized to understand
and  control the impact of any specific chemical on the environment.   The rates of
transfer between  media,  the  capacities  of  the media,  and  the  mass  transfer
coefficients need to  be defined.   In some cases, the kinetic measurements have not
been attempted because of the difficulty of  monitoring the mechanism.  Significant
environmental  transport  parameters in each medium, (e.g., participates, soils,  and
sediments) must be  characterized in terms of impact on the  mechanisms of  the
transport processes. Distribution of the specific  chemicals on surfaces or on particles
of different sizes must also be examined. All these parameters must be characterized
for TCDD and all dioxins and furans.

1.4.3 Advection/Diffusion/Dispersion

      Advection  of  a chemical   in  air, surface  water,  and interstitial water in
groundwater systems is the movement of the chemical with the bulk movement of the
air or water.  In some  environments, chemical phases also need to be  considered,
including  two-phase  flow  (water/organics),  suspended  participates,  and  colloids.
Although characterizing  the advection of vapors and dissolved chemicals in water is
frequently  difficult, predicting advection in a second phase presents a more  serious
problem that remains unresolved in many instances.

      The state of the art for estimating rates of diffusion and dispersion was ranked
at previous  EPA workshops  on  exposure assessment  as  poor to fair,  and  current
estimations of diffusion and dispersion rates for  TCDD isomers were ranked as poor.
In general,  molecular diffusion rates are  calculated  using estimates of molecular
diffusivities.  Procedures for estimating molecular diffusivities  in the air are the most
accurate; procedures for estimating diff usivities in water and soils  are less accurate.

      In addition to estimation of diffusivities,  other  physical parameters must be
either measured or estimated to calculate  diffusion  rates in soils and sediments.
These parameters include void fraction of soils or sediment volumes and tortuosity of
the diffusion path.

      Dispersion of chemicals in  air and surface waters results  from the turbulent
motion  of the fluid.  Dispersion rates in  these media  are functions of a variety of
physical parameters such as  wind or water  velocity,  and temperature  and density
                                       19

-------
gradients.  In addition to possible turbulent motion, dispersion may occur  because of
the heterogenous characteristics of solid phases. Consequently, dispersion in soils and,
potentially, in sediments must take into account site-specific characteristics and scale
factors.

Research Needs

      Calculations of the amount and rate of TCDD adverted into, through, and out of
each medium are required for both the  vapor phase and TCDD sorbed to participates.
The need  to  improve abilities to characterize,  measure, and  predict advection in
saturated and unsaturated soils, sediments (interstitial waters),  and  estuarine waters
should receive  the  highest  priority.  Also, advection in two-phase flow  must  be
characterized for these systems.

      The rates and extent of the diffusion and dispersion processes for TCDD must be
better characterized  for  saturated and  unsaturated soils  and  sediment  systems,
including the two-phase flow. This also includes the development of better techniques
for measuring the significant environmental factors that govern the process rates  and
extent, such as void fraction and soil temperature.

                                 1.5  Modelling

      For the transport and transformation  of individual chemicals in air,  water,  and
soil  environments,  many  mathematical  models  are  available  to predict  exposure
profiles necessary for  bioavailability estimates.   Models offer a method to  organize
information on many different processes  occurring simultaneously, and facilitate  the
interpretation of  laboratory  and field  observations and the determination  of  rate-
controlling steps.  Models range from  very simple, steady-state  algorithms to very
complex, process-oriented  dynamic  codes.   Simple models are  easy to use but  are
usually inappropriate for site-specific field  situations.  More comprehensive, complex
models, although possibly more realistic, may contain variables for which data are  not
readily available  or for which methods  are inadequate  or  unavailable to  measure
magnitudes in time and space.  Examples of this problem include measuring the water
conductivity  in  soils and  the spatial variability of dispersion  coefficients in air  and
soils. However, judicious selection and use of appropriate models can  provide  the most
reasonable approach for solving complex exposure problems.

Research Needs

      With regard to the use  of complex models, there is a continuing need to develop
more efficient  numerical  methods  to  solve coupled  systems of nonlinear, partial
differential equations that  are stable and present  solutions with  low levels  of
numerical dispersion. To facilitate the  development of realistic complex models,  the
efforts of  the laboratory scientists, field researchers, and modellers must be closely
coordinated.  Without such coordination, inappropriate models may be developed and
applied to field conditions to predict  exposure  concentrations from contamination
episodes.
                                       20

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     In addition, it  is  necessary  to determine  the conditions  under which  single
chemical models  can legitimately be used to predict the transport and transformation
of mixtures.  It is tacitly assumed that for contaminants with low solubilities, and even
for most designated air pollutants, the concentrations in the transporting medium are
so low that they act independently of one another.  This assumption is certainly not
true in a photochemical smog episode,  and it remains  an issue  for which no clear
criteria exist to determine the set of chemical properties or concentrations for which
interactions are  significant or negligible.   Thus,  models that address the  issues of
mixtures and their  interactions are needed, particularly for dioxins and furans. Also,
the application of models to the complexities of multiple-phase transport imposes an
additional level of difficulty in developing meaningful numerical solution techniques.

                   1.6 Analytical  Methodology For Analyses Of
                   TCDO In Environmental And Human Samples

     Analytical  methodologies currently utilized by EPA  for  analyzing TCOD in
environmental and human samples involve:

1)   fortification of the sample with an isotopically labeled TCDD;

2)   extraction with an organic solvent when analyzing soil, sediment, and water; or

3)   saponification with  alkali followed   by  solvent extraction when  analyzing
     biologicals such as human tissue.

     Interferences are  removed by  acid  and base partitioning  steps, followed  by
chromatographic cleanup on alumina and charcoal  columns or high performance liquid
chromatography. A portion of final concentrated extract is subject to isomer-specific
analysis by capillary gas chromatography and mass spectrometry, using selected ion
monitoring.  The quantity of  TCDD reported is corrected for  recovery efficiency of
the labeled standard in each sample.

     This analytical methodology has focused on TCDD; however, it  has also been
extended   for   the  quantitative  determination   of   other   dioxin  isomers  and
polychorinated  dibenzofurans.   Several matrices  have  been analyzed,  and their
respective minimum detection limits (MDL) are listed in Table 1.2.

     Rigorous   quality  assurance  has  been  incorporated   into  the   analytical
methodology.  Samples are  analyzed  in well-defined sets that include  blanks, spiked
matrices, duplicates, and blind samples.  Some samples are exchanged and analyzed by
outside laboratory collaboration.  A panel consisting of scientists  outside of EPA has
been established to review analytical results before their release.

Research Needs

     Sufficient supplies of TCDD are needed for research, as well as for standards and
reference samples for quality assurance and quality control.
                                       21

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                                   Table 1.2



              Minimum Detection Limits (MDL) of Selected Matrices3
Matrix
Water
Human milk
Deer adipose muscle, liver, bone marrow
Human adipose, beef adipose, beef liver
Elk adipose
Pork adipose
Pottery clay
Dog adipose
Fish, herring gull tissue
Soil, sediment
Fly ash from coal-fired power plants
Fly ash and gas-phase effluents from municipal
incinerators
Chemical disposal sites
Chemical products and processes
Chemical destruction processes
MDL
0.008
0.1
0.4
0.5
1
h
h
1
1
1
1

1
10
20
20
Range
to
to
to
to
to
to
to
to
to
to
to

to
to
to
to

i.oi
5
5
10
5
7
7
10
10
10
10

300
800
800
800
a  Prepared by A. Dupay and R. Harless.




b  Based on 1000-g samples.
                                      22

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             Environmental Processes in Bioavailability Working Group



                               PARTICIPANTS
Jimmy Boyd



Peter Chapman



David Cleverly



Donald Crosby



Paul DesRosiers



Aubry Dupuy



Walter Farmer



Virgil Freed



Randy Freeman



Robert Harless



Takeru Higuchi



Greg Kew



Christopher Kouts
Douglas Kuehl



Bill Lamason



John Loper



Leland Marple



Arthur Martell



Danny McDaniel



Jay Means



Theodore Mill



Warren Piver



John Quenson



Michael Roulier



Walter Saunders



Jerry Schroy



Terry Stoddart
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                                  CHAPTER 2

                       BIOAVAILABILITY IN ECOSYSTEMS

                 Co-chairpersons:  Thomas Duke and Mark Harwell


                                2.1 bitroduction

      Chlorinated hydrocarbons, including dioxins, are found in terrestrial and aquatic
ecosystems and are bioavailable to certain  organisms.   The bioavailability of these
chemicals  should be considered in a holistic manner,  because ecosystems typically
consist of  dynamic assemblages of  thousands of species existing within a complex and
heterogeneous  chemical  and   physical  environment,   with  which  they  interact.
Biological  organisms control ecosystem processes of energy and material flow, and
they modify the chemical and physical  environment. Alternatively, the environment
substantially affects the distribution, abundance, and diversity of these biota.

      An important aspect of ecosystems is that they operate simultaneously on widely
differing spatial, temporal, and structural scales.  The  focus may  be  on  individuals,
populations, and communities of species, or on integrative measures such as diversity,
productivity,  respiration,  and  decomposition.    For our  purpose  here,  the  term
ecosystem  includes all of  these aspects, and  the  term bioavailability  means the
exchanges  between the environment and the biota, and exchanges from biota to other
biota.  In  large  part,  this mutual interaction  between the environment and biota
focuses on  the interface between chemicals in an abiotic  phase and between chemicals
within the biota.   Exchange  across  this  interface  is  the  central element  of
bioavailability.  The concept  of bioavailability here includes both  the dynamic and
steady-state aspects.

      Both  the rates of exchange,  such as the  rate of dioxin uptake by plants  from
soils, and  the ultimate (steady-state) levels of  dioxin  accumulated in biota are of
interest, especially as represented  by  bioconcentration factors  (e.g.,  the dioxin
concentration in plant tissue divided by the concentration in the soil).

      The ecosystem considerations for dioxin have two distinct facets:  the effects of
dioxin on the biota in the ecosystem, and the role of the ecosystem in mediating dioxin
exposure to humans.  The  human exposure pathways represent  a special case of the
more general  investigations of  the fate  and transport  of dioxin  within ecosystems.
Routes that take dioxin from an abiotic phase into biotic material ingested by humans
are worthy of careful  attention;  these are  discussed  in the following sections  on
aquatic and terrestrial ecosystems.

      Effects on ecosystems begin  with  effects on individuals.  These effects can be
manifested as physiological  responses, behavioral responses, or even  death  of

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individual organisms. But translating effects on individuals to effects on a population
is  not  a  simple  extrapolation.   Effects  on populations  are influenced  by the
interactions of the individuals within the population and with other species, and also by
the  nature  of  the  physicochemical  environment.   For   example,  compensatory
mechanisms  may  decrease  adverse effects  on populations; conversely, differential
sensitivity within life stages may result in greater effects on natural populations than
would be predicted from laboratory bioassays on individuals.

     Community-level  effects  involve  an increase  in  complexity,  where indirect
effects may result when a population not directly exposed to dioxin in the biotic phase
becomes indirectly exposed through food-web interactions.   Community composition
can  be  affected  by indirect effects  involving predator-prey interactions, changes in
competitive relationships, and induced susceptibility to disease or parasites,  among the
many various possible interactions.  In  considering community compositional changes,
particular species or groups  of species may be identified as being especially important,
whether for economic, aesthetic, or ecological reasons.

     Finally, additional factors  influence  ecosystem  process-level  effects.   For
example, if different species perform the same functional roles, whole species can be
lost  or  populations altered  without changing ecosystem processes,  because of such
functional  redundancy.  Conversely, if fundamental ecosystem processes are affected,
biological changes can  be  expected to occur in commmunity  composition,  which  is
dependent  on those processes.  Thus, in some instances,  biological responses to toxic
chemicals  might  be  better  understood by measuring ecosystem  processes directly
rather than by monitoring species composition and diversity.

     To predict effects of dioxin on ecosystems and on humans, the transport and fate
of dioxins  in the environment must be understood.  The bioavailability of dioxins to
biota, exchanges among biota, and influences of  organisms on the movement of dioxins
within and across systems  must be included.  A conceptual model of exchanges of
dioxins among ecosystem components appears  in the next section.  Subsequent sections
address transport  and fate in aquatic and terrestrial ecosystems, effects  on  biota
(species and processes),  and  pathways to humans.

     In these sections, three  sets of  topics were considered:  1)  identification of
ecosystem processes  that are involved  in routes, rates,  and reservoirs of dioxins in
aquatic and terrestrial  ecosystems, sensitive to effects of dioxin contamination, and
involved in  biological  decontamination  processes;   2)  identification  of  particular
species and communities that are potentially affected by dioxins; and  3) the role of
food chains and food webs in human exposure and risk.

     Overlying all of these  topics are issues of  scale across  time and space and issues
of uncertainty.  Scale issues include  determining  the spatial and temporal extent of
exposure and effects, differentiating chronic versus acute exposures, and considering
whether there is sufficient  time for  potential compensatory mechanisms to operate.
Uncertainty follows from lack of information, spatial and temporal heterogeneity that
can  obscure  effects, differences  among species  in sensitivity to  direct  effectj,
propagation of indirect effects, and unexpected consequences.
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              2.2  Exchanges Of Oioxin Among Ecosystem Components

2.2.1  Conceptual Model

      The purpose of  most ecosystem  models is mathematically to describe complex
ecosystem interactions among  biotic and abiotic components.  Once an appropriate
model has been  developed,  it can  be used to answer  questions about  higher-level
processes, such as chemical  transfers through the ecosystem.  Information on  both
process rates  and component concentrations is utilized.  Mathematically, it becomes
more  difficult to develop and verify a model  which increases  in  complexity from
species to population to community  level.   At the ecosystem  level, validation  of
models is difficult or impracticable.

      To obtain a. better understanding of the bioa vail ability of dioxins to both lower
and higher forms of  life, and movement  from one  component  to another, and  to
identify  areas of  critical research  needs,  we  have presented  a food-web  model
applicable to  both aquatic and terrestrial  ecosystems (Figure 2.1).  The conceptual
model figuratively describes interactions  and dioxin exchanges  among abiotic and
biotic components.  The dioxin pools  in a number of biotic  components are  shown,
including the transfers among biotic compartments through feeding.  Also shown is a
general abiotic pool of  dioxins.  The open arrows represent the exchanges of  dioxin
among abiotic phases  and each type of  biota.  Details can be added  to this abiotic pool,
making the conceptual model  applicable to different types of ecosystems.

      The model is generic; from it,  calculations can be made based upon assumptions
of the thermodynamics of tne toxicant.  We  recognize that equilibrium conditions  do
not always exist in the environment;  however, we believe this is a  useful approach for
calculating initial  estimates of  dioxin  concentrations  for some components.    In
general,  the tendency for change is towards thermodynamic equilibrium.  Thus, the
bioavailability of dioxins to  organisms can be evaluated,  and the hazard or risk  of
compartmental concentrations can be assessed.

2.2.2  Research Needs

      Research should be conducted to:

      •  measure concentrations  of  dioxins with time in organisms as a function  of
         dose in food, water,  and other sources for model production;

      •  use microcosms to verify models; and

      •  conduct  a full-scale ecological study at a highly contaminated site.  This
         should include field studies of  fate, chronic effects, and ecological processes,
         with supporting laboratory studies, and studies of the mechanisms of effects.

                     2.3 Bioavailability: Aquatic Ecosystems

2.3.1  Introduction

      The distribution, transport, and fate of TCDD and other dioxins and  furans  in
aquatic ecosystems are governed  by  processes that  are generally known from studies

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                                Dioxin Pool Food Web Model
                Primary
                Producer
                 Dioxin
                 Pool
 Primary
Consumer
 Dioxin
  Pool
                                     Macro-
                                   invertebrate
                                    Microbial
                                   Decomposers
                                     Dioxin J
                         Abiotic
                         Dioxin
                          Pool
                                                                    Secondary
                                                                    Consumer
                                                                     Dioxin
                                                                      Pool
Figure 2.1     Dioxin pool food-web model.  Solid arrows indicate abiotic dioxin flux
               by predation and feeding; open arrows indicate direct exchange between
               the abiotic and  biotic components.  This concept  can be expanded to
               demonstrate dioxin pool exchange between two  more  foci webs of the
               ecosystem.
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of other hydrophobia organochlorine compounds such as PCBs.  All these compounds
share an affinity for the carbonaceous component of suspended and settled sediment
and  for dissolved organic matter; these characteristics influence bioavailability.  Less
can  be inferred from these studies when we attempt to predict biological  effects of
chlorinated dioxins  and furans because of  uncertainties  that exist  with  regard to
uptake, metabolism, and the toxic model of action of these compounds.  For example,
the selective bioconcentration of TCDD observed in fish exposed to mixtures of dioxin
isomers  has not  been predicted  on  the  basis  of dioxin isomers1  similar water
solubilities.

      Our  ultimate  research   goal  is to develop  methods  for   predicting  the
bioavailability of dioxins and furans to aquatic organisms in different environmental
settings  on the basis of the physical and  chemical properties of isomers, routes of
exposure,  water chemistry, sorbent  interaction, and  pharmacokinetics of  uptake.
Estimates  of  the  potential environmental  impacts of  dioxins  will  be improved and
strengthened if the dose received by organisms can  be accurately predicted.  From a
human health perspective, prediction of the amount of contaminant accumulated by
organisms  can indicate whether unacceptable levels may be expected in  animals
consumed by humans.   Development of this capability will thus aid in designing water
quality  criteria and in evaluating  the  environmental risks associated  with  existing
contamination and with future releases.

2.3.2 Routes and Rates of Uptake, Metabolism, and Elimination

      The bioavailability and accumulation of  a contaminant depends on its  physical
and chemical form and  the rates at which it is  taken up, metabolized, and eliminated
by organisms.  Physicochemical partitioning within  the aquatic environment exerts a
major influence on bioavailability of dioxins because the rate at  which a compound can
be taken up by  an organism will vary widely,  depending on the physical matrix  with
which it is associated.  By analogy with other organic compounds, dioxins dissolved in
water are  expected  to  exhibit  a high rate of uptake. Contaminants associated  with
organic and inorganic particles, dissolved organic matter, or food appear to be less
readily incorporated. The rates of  dioxin uptake from different source compartments
and the physical and biological  factors that affect the rates of incorporation  need to
be examined  quantitatively so that  the  total incorporation  by organisms can  be
predicted.

     Although dioxins, like many other organochlorine contaminants, do not appear to
be readily bio transformed, the rates and pathways of detoxification by  aquatic
organisms  need  to be  examined.   Information on  the  pharmacodynamics  of dioxin
distribution among tissues within an organism is needed to understand  the metabolic
fate, the mechanisms of toxicity, and the accumulation in tissues that are  consumed
by humans. Development of correlations between  tissue distribution of dioxins and
molecular  components  of the  tissue,  such as lipid content,  would be  particularly
relevant.

     Rates of  elimination of TCDD  appear to be  very low,  although other TCDIJ
isomers may be eliminated more  rapidly.   Because the  body  burden of  dioxins
accumulated by  organisms will be largely determined by their elimination rates,  it is
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important to increase our  knowledge about  the rates  and routes of elimination of
dioxins.

     The rates and routes of uptake, metabolism, and elimination of dioxins, and thus
the body burden present at  any time, will be  affected by natural and human-induced
changes  in  the environment.    The effect  on  these  processes,  of environmental
variables, such as temperature,  salinity, water  chemistry, and  dissolved oxygen, as
well as the effects of nutritional status of  the organism  must be  understood if a
predictive capability is to be achieved.

2.3.3 Effects of Dioxin on Ecosystem Processes

     Toxic  organics can alter ecosystem  processes.   Although there  are no data
directly  implicating  dioxins in  altered  ecosystem  processes,  there are sufficient
toxicological data to warrant concern. From the known  physical chemistry of dioxins,
it is expected that  dioxin effects on ecosystem processes will be similar to effects of
other toxic organics for which data exist (e.g., PCBs and  PAHs).

     Productivity,  both primary  and   secondary,  is   a fundamental  process  of
ecosystems.  Dioxin contamination may  pose  a risk to the balance of productivity.
Should dioxins  affect  any functional group of organisms (e.g.,  phytoplankton,  filter
feeders, or carnivores), there could be a shift  in the energy flow patterns that might
be detrimental to the overall balance of the ecosystem.

     At  the ecosystem level,  the processes most sensitive to  dioxin toxicity would
likely be alterations of population behavior, fecundity, immunological resistance, and
other life-history  characteristics.    However,  the  classic  approach to  community
structure analysis is too coarse a measure. By the time parameters such as diversity
are affected, the system is usually heavily contaminated.

     There are insufficient aquatic field data on effects of dioxins.  A data base needs
to be developed from which the long-term consequences of dioxins in the environment
can  be evaluated.  Research should focus on the key ecosystem processes described
above.

2.3.4 Biological Decontamination Processes

     Mechanisms to eliminate dioxins from ecosystems may have both biological and
physical  components.   Biologically,  microorganisms have the  potential  to degrade
dioxins.  Physically, photooxidation may be important in aquatic environments,  but the
removal  of dioxins from  the  biologically active  zone  by  sediment  accumulation
processes and burial is likely  to be more important.  Because dioxins are associated
with fine-grained  sediments,  the transport and long-term fate of  dioxins will  be
mediated by sediment  transport processes.  We also need to understand the  dynamics
of sediment transport as a mechanism for remobilization.

2.3.5 Biological Effects

     Toxic  effects  of dioxins and furans in actual aquatic  ecosystems have been
studied in only a few instances. Factors complicating such studies include the  virtual


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absence of useful exposure data, particularly for water concentrations, the presence of
complex mixtures of many organochlorine compounds, the nonspecific symptomology,
the slow mode of action, and the levels of accumulation.

      Fish-eating birds  can  accumulate  TCDO and related isomers.   Reproductive
effects,  including  embryotoxicity  and  teratogenicity, are consistent with  early
research  on  the  chicken  and  the  putative  agent in  chick   edema   disease,
hexachlorodibenzodioxin (1,2,3,7,8,9-HCDD).   While previous studies  on  Lake Ontario
herring gulls are now artifactual, unpublished research on the Forster's  tern on Lake
Michigan show continuing reproductive problems consistent with the known effects of
these compounds.

      The available  data describing TCDD toxicity to aquatic organisms are limited to
fish.  Laboratory exposures of fish to TCDD that have been reported can only be used
for a gross estimate of toxic effects. No information exists on the toxicity of other
dioxin or furan  isomers.  In separate  studies, northern  pike (eyed eggs) and rainbow
trout (juveniles) were exposed for four days to a range of static TCDD concentrations
and held for  depuration in noncontaminated well water. These studies  indicate that
TCDD is a  slow-acting toxicant.   However, the mode of toxicity appears to  be
different than  for  most other  neutral lipophilic organic chemicals.  It is  not clear
whether  short-term  exposures  can  ultimately  produce  the same  dose-response
relationships as continuous exposure to the same or lower levels over longer periods of
time.  The mode of  action of TCDD toxicity in aquatic organisms is unknown.

      Traditional methods for assessing acute toxicity, such  as the 96-hour LD5o> are
not useful in dioxin hazard evaluations.  The available data are insufficient  to gauge
the magnitude of variability in interspecies sensitivity, although this  may  be quite
large, as is  the  case  for mammalian toxicity studies.   At this  time, a no-effect-
concentration cannot  be determined.  Growth  and  reproduction  have  not been
investigated  for fish or invertebrates exposed to trace  concentrations  of TCDD for
sufficient periods of time to  allow steady-state concentrations to be reached  in
tissues.   Bioassays  performed  with  complex  mixtures  of  dioxins, furans, and other
organochlorine compounds produce toxic effects  that indicate additive and possible
synergistic interactions.

      The toxicity of TCDD  associated with suspended or settled sediments has  not
been  determined.   Water  TCDD  concentrations resulting  from   partioning with
sediments may affect toxicity,  especially if organism  uptake is primarily via  the gills.
The  ingestion of contaminated sediments, particularly  by  bottom-feeding  fish and
benthic organisms,  presents a  potentially important  alternate route for uptake and
consequent  toxic effects.   Bioassay systems adequate for  long-term,  controlled
exposure of aquatic organisms to suspended and settled sediments are needed.

2.3.6 Role of Food Chains in Human  Exposure

      Aquatic organisms and  fish-eating  avians may  accumulate dioxins and related
compounds to levels of concern relative  to  both toxic  effects and risks to  huma i
consumers of the organisms.  However, a comprehensive understanding of dioxins that
would allow  the prediction of body  burdens  from exposure data has  not  yet been
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achieved.  Microcosm  studies have provided estimates for bioconcentration factors
(BCFs) for aquatic piants and animals; however, measurement of steady-state BCFs
and those concentrations that may be achieved in nature have not been undertaken.
Relationships between sediment concentrations and fish tissue concentrations are open
to interpretation. Data suggest that fat or fat content of individual tissues may be an
important determinant of bioconcentration in aquatic species.  In summarizing  the
uncertainties related to body burdens  in aquatic organisms,  it appears that  age, size,
species, feeding relationships, and fat content are key considerations from a  biological
standpoint.   Abiotic factors  probably include:  sediment transport and  availability;
characteristics such as particle size, organic content, and general complexing ability
by organic compounds; and solvents as  dissolution agents.

      Sufficient  information   is  available  to   establish  the  bioavailability   and
bioconcentration of TCOD and related compounds in salmonids and other species in the
Great Lakes, in rivers and ditches that drain dumps, and in fields sprayed with 2,4,5-T.
Knowledge of dioxin kinetics is  much more sketchy concerning estuarine and marine
species.   Fish,  snapping  turtles, and fish-eating birds have been  characterized in
individual  situations,  but  there  is little  understanding of the   potential  for
biomagnification along food   chains  within  a  contaminated  ecosystem.    Because
biomagnification is associated with orders-of-magnitude increases across biotic media
boundaries, understanding the  potential for biomagnification can be best determined
by placing special emphasis on birds and mammals feeding on aquatic organisms.

2.3.7  Research Needs

      The key area to  be  addressed in future research is a better understanding of
factors controlling bioaccumulation.  In addition to laboratory  research on  individual
aspects such as fat  content and partition coefficients, full abiotic/biotic food-chain
research should be conducted to characterize contaminated areas.  Areas differing in
the relative  dominance of perceived factors  affecting food-chain transfer of dioxins
and related compounds should  be  studied.   To understand  and develop a total human
exposure model and  institute  risk assessments, aquatic food-chain influences (e.g.,
fish, turtles,  crayfish, and waterfowl) need to be integrated with terrestrial wildlife
concentrations,  domestic  food  intake,   and   occupational/ambient   exposures   to
determine adequately the primary pathways to humans.

      Research should be conducted to:

      •   develop the capability  to predict dioxin  levels in tissues (particularly in
         organisms that constitute human food chains) as a function of environmental
         conditions;

      •   develop data for  understanding the  mechanisms of toxicity and the factors
         responsible for differential toxicity among species; and

      •   determine the nature and extent of disruption caused  by  dioxins in aquatic
         communities and the  mechanisms by which they are caused.
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                    2.4  Bioavailability: Terrestrial Ecosystems

2.4.1  Introduction

      Few acute  and chronic toxicity  studies have been conducted  to  study  TCDD
effects in the field.  The few field studies conducted on wildlife that are reported in
the literature involve sampling  trophic levels at contaminated sites and determining
TCDD concentrations. Very few laboratory studies have been  conducted to study the
acute  and chronic toxicity  of  TCDD  to wildlife as well as effects on  ecosystem
processes.  The limited  data that are available from  controlled laboratory and field
studies show  that TCDD bioaccumulates in terrestrial wildlife  up to about 25-fold and
that bioconcentration among trophic levels does not occur to  any significant extent.
In areas containing high concentrations  of TCDD, studies are needed concerning TCDD
bioaccumulation,  bioconcentration in trophic levels, and ecosystem process effects.  In
isolated cases, some risk to humans could occur from food animals that have  direct
access to contaminated soils. The risk is most significant when individuals consume
their  own farm-raised products.  Risk to  the general public would  be  smaller because
of dilution through marketing channels and diversity of dietary sources.

      The major routes of contamination in terrestrial ecosystems are likely to  be the
movement of dioxins from soil to  animals and from the  atmosphere to plants.  Liver
and adipose tissues are  the most likely reservoirs for dioxins  in animals, and those in
intimate  soil contact  harbor  the  highest dioxin  concentrations.   Assuming  that
susceptibility will  be largely  a function of exposure,  those animals  ingesting  or
inhabiting soil are the most  likely to be affected by TCDD.   This  includes domestic
animals or wildlife that eat soil along with food or grooming, as  well as burrowing
animals  such  as groundhogs,  rodents,  and  rabbits.    Soil-dwelling  micro-  and
macroinvertebrates are also likely to receive high exposures.

2.4.2  Degradation of TCDD in Soil

      The major  processes of  TCDD  removal  from  soil are photodegradation  and
volatilization.  It  is generally agreed that microbial degradation of TCDD occurs  to a
very limited  extent.  Preliminary results of microbial degradation studies underway in
Missouri  indicate  that microbial degradation occurs, but it is  too early to draw firm
conclusions.    Refer to Chapter   1  of  this report,  Environmental  Processes  in
Bioavailability, for additional information on this subject.

2.4.3  Bioconcentration in Wildlife from  Soil

      Whole-body  and tissue concentrations of TCDD have been measured in a variety
of  wildlife   species  under  conditions  of  chronic field  exposure.    The highest
concentrations of TCDD are generally found in the liver and adipose tissues. Although
dioxin concentrations may be influenced by factors such as trophic level, fat content,
physiological  state, and sex of the organism, the primary factor influencing uptake by
wildlife appears to be the degree of contact with contaminated soil.  In earthworms,
which continually consume and contact  soil, a  direct linear correlation has been  shown
between  body  burdens  of TCDD  and  soil concentrations.    Despite reported body
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burdens of TCDD in  numerous organisms, no studies have definitively shown either
population or community effects.

     Unlike  aquatic  animals,  terrestrial  animals do  not  concentrate TCDD to a
considerable extent. BCFs are generally around  25 or less.  The highest BCF reported
is 42, found in earthworms in Seveso, Italy.  Interestingly, absolute concentrations of
TCDD  in aquatic animals have not exceeded those found in terrestrial animals despite
the high propensity for  movement from the water  column into aquatic animals.  It
should  be noted  that BCFs for aquatic organisms usually describe the partitioning of
dioxins between water  and organisms,  whereas BCFs for  terrestrial animals usually
describe the  partitioning between  soil  and organisms  that may not  be in  constant
contact with soil.

     There may be potential human food-chain exposure from  wild terrestrial birds,
such as wild  turkey,  pheasant, quail, and  grouse, but  no data  exist to substantiate
whether TCDD and related compounds  (e.g., furans) exist in  these species.  Turkeys
seem to be more sensitive to contaminants such as PCBs than the other species.
However, relative sensitivities among species  may  be important to  humans because
wild birds may continue to accumulate concentrations of dioxins until consumed for
food.

2.4.4 Movement through Soil to Food Animals to Humans

     The major  route  of potential TCDD exposure to farm  animals is by  direct
ingestion of soil. Intake of soil by grazing  cattle and sheep is inversely related to the
amount of available forage and may range from 2 to 15% of dry matter intake. Cattle
may also consume soil at 2 to 4% of dry matter intake when confined to lots with no
vegetation.  Pigs consume 2 to 3 times as much  soil as cattle.  Data are not available
for  soil consumption by poultry with access  to soil.

     Monitoring data show a general correlation between herbicide  application  rates
and TCDD levels in cattle.  It has been suggested that BCFs for TCDD from soil to
mammals range  from  2 to 25.   Field  experience  with  the chemically  related
polybrominated biphenyls suggests that this is a reasonable conclusion for nonlactating
ruminants such as cattle and sheep.  This factor is slightly lower for lactating cattle
because of the secretion losses  through milk.  BCFs are several times higher in pigs
because of higher soil consumption. No BCFs are available for  poultry.

     TCDD is known to be highly toxic to  domestic poultry, but essentially nothing is
known  regarding the quantitative  aspects of   dioxin  interaction with these birds,
particularly as related to potential metabolic pathways and residue retention by edible
tissues or secretion into eggs. However, because of the highly controlled environment
in which most commercial poultry is raised, soil contamination by dioxins is unlikely to
result  in significant  concentrations in  commercial  poultry.   In  barnyard  poultry,
exposure at dioxin-contaminated sites is certainly possible, and limited studies suggest
that contamination of meat (and presumably eggs) would result.  There are no data to
indicate what  levels of dioxin residues in poultry meat or eggs would be expected from
a given dietary intake level.
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      Given the  nature  of  the presently  known  and likely to be  discovered dioxin-
contaminated terrestrial sites, the major potential for human  dietary  exposure to
dioxin residues would probably occur  through the consumption of meat or milk from
farm animals held on contaminated  sites.   A number  of available  studies clearly
indicate that exposure  of  farm animals,  particularly  cattle, to dietary dioxins will
result in residue retention by edible tissues and secretion into milk.

      Studies with the higher chlorinated  dioxins suggest that in livestock, a limited
degree  of bioconcentration in tissues over  dietary  levels  may  occur.     But,  if
bioconcentration occurs with TCDD, it may be considerably less.  The available data
on dioxin interactions with both lactating and beef cattle appear to  be  only  marginally
definitive with respect to their value  in predicting residue levels that would occur in
meat or  milk as a result  of specific levels of dietary exposure.  Lactating cattle fed up
to 500 ppt dietary TCDD  resulted in  milk levels  of approximately 100 ppt, although
24 ppt dietary TCDD to beef cattle  gave up to  100 ppt in fat  and 10 ppt  in liver.
Studies have  demonstrated that normal cooking  processes do not generally reduce
dioxin levels present in raw  meats.

2.4.5 Movement  through Soil to Plants

      TCDD  can  translocate  through  plants; however, uptake from soil  is generally
very small, with  an extremely small accumulation of dioxin into  fruits and seeds. In
field  studies, contamination of aboveground plant parts  has often  been attributed to
contaminated dust.

      Dioxin  binds to organic surfaces, including plant  roots, and  in the bound form is
unavailable for absorption and translocation.  Concentrations in fleshy roots at the
root surface  typically are similar to soil concentrations, with a sharp decrease toward
the root  center.

      Uptake from solution  and translocation into  aboveground plant tissues occurs at
rates that are easily measured and  would  be  of concern under  some  conditions.
Concentrations in soybean and oat leaves have been shown to be five and ten times
(plant dry weight) the solution concentration after one day of exposure.

      A  possible  contradiction between field  studies and some  laboratory  uptake
studies probably results from imprecise descriptions of the root/soil  solution interface.
In soil, dioxin is  normally bound to  particles and is therefore unavailable for uptake.
Even binding to exterior root surfaces makes dioxin unavailable for  uptake.  However,
when dioxin is in  solution at the exterior root surface, uptake and translocation occur.
Therefore, conditions that change dioxin solubility in soil may facilitate plant uptake.

      There is no evidence in the scientific literature  of  plants  being  affected by
dioxin, even at the highest levels of  contamination studied.  However, specific studies
to examine phytotoxicity have not been done.

2.4.6  Research Needs

     Research should  be conducted to:

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•  evaluate  the  chemical  and  biological  characteristics  of  residue  from
   experimental incineration projects and apply the results to risk assessment;

•  identify the mechanisms by which TCOD in bound forms is released in the gut
   and taken up by lactating and food animals; and

•  determine the effects of soil organisms and plant roots on vertical transport
   and bioavailability of TCDD.
                                 35

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             BIOAVAELABELITY IN ECOSYSTEMS WORKING GROUP
Brian Butterworth



Philip Cook



Robert Diaz



Tom Duke



George Fries



Mark Harwell



Robert Huggett



Wayne Ivie



Darcy Johnson



Kenneth Johnson



Steve Kennel



Tim Kubiak
                              PARTICIPANTS
Douglas Kuehl



Ray Lassiter



John McCarthy



James Craig McFarlane



Hap Pritchard



Thomas Sabourin



Richard Tucker



Barbara Walton



Armon  Yanders
                                    36

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                                   CHAPTER 3

                         BIOAVADLABILITY TO HUMANS

                Co-chairpersons:  Diane Courtney and Michael Gallo


                                 3.1  bitroduction

      This  work has  focused on  bioavariability to  humans,  which refers to  those
characteristics of the toxicant  (e.g.,  its form,  route, matrix, and concentration) and
the  host  that determine the  internal  biologically active  dose.    Risks  may  be
inaccurately estimated  in  the absence  of knowledge  about factors  determining
bioavailability, even when exposure is relatively well defined.  Clearly, matrix and
route effects are likely to be significant.  However, human responses and  risk are also
influenced by exposure and by differences in the sensitivity of target  sites of action.
To  consider bioavailability  adequately,  exposure  and toxic  response must also  be
examined.

      TCOD was the main concern of this report. The biological effects of TCDD have
been  more  extensively   studied  than  any  other  chlorinated  dibenzodioxin  or
dibenzofuran.  However, because many isomers of dioxin are usually found at the same
sites, the possible influence  of  other  isomers of dioxin on the bioavailability of the
TCDD isomer is well recognized, and  workshop discussions could readily be applied  to
these other dioxin isomers as well.  Additionally, dibenzof uran isomers are often  found
in  the  same  sites  as  the  dioxins,  and  the furans  could easily influence the
bioavailability of TCDD and the other dioxin isomers. Because the furans and dioxins
have  very  similar toxic end points, it was  recognized that the following discussion
would apply to tetrachloro-, pentachloro-, hexachloro-, and  heptachloro- dioxin and
f uran isomers.

      Bioavailability to humans, as the host organisms, has two aspects:

      1)  the uptake of chemicals from environmental matrices into the host and the
         interactions with critical receptors and tissues within the host; and

      2)  the subsequent  distribution,  redistribution,  or mobilization within the host.
         Mobilization  is a means  of  internal in vivo bioavailability that  could  be
         beneficial  (by promoting excretion) or detrimental (by increasing exposure  to
         critical organs).

      These two  aspects of bioavailability to humans served as  the definition  of
bioavailability  for  this  workshop  group.    Additionally, it  was  recognized  that
bioavailability could be an index of  the potential for incorporation into humans.
                                       37

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                     3.2 Bioavailability Of TCOO To Humans
                          From Environmental Matrices

3.2.1 General Properties of TCDD and Matrices

      Physical and chemical properties of TCDD that should be considered include
partition coefficients, vapor pressure, lipid coefficients, and solubilities, which  have
been discussed in Chapter 1.

      Matrix types and  characteristics that should be considered include soil, fly ash,
aerosols, and solutions.  A recent finding is that the length of time the TCDD is bound
to soil may be critical for the subsequent extraction of TCDD from the soil, in that
the longer the time TCDD is bound to the soil, the harder it  is to extract TCDD.
Various soil  types result in distinctly different  strengths of chemical binding  with
TCDD.   Bioavailability is primarily dependent on  the binding of  TCDD to different
matrices.

3.2.2 Research Needs

      Matrices of various composition with known TCDD concentrations should be used
to determine the bioavailability of TCDD by using the same species and toxicologic
end  points.  Soil  should be considered as a priority matrix because it is the most
common  one  to which humans  are exposed. Contaminated fly ash  or  respirable
particles should be studied to determine  their TCDD bioavailability to humans.  A
range of concentrations should be utilized, because the bioavailability of TCDD may
differ at differing concentration levels.

                            3.3  In Vivo Bioavaiiability

3.3.1 Mobilization and Redistribution

      In vivo bioavailability is a measure of the amount of TCDD that is at the target
organs, cells, or cellular constituents.  In addition to  the TCDD that  is available from
environmental  matrices, TCDD is also  bioavailable from  TCDD  mobilization  or
redistribution within the host in specific tissues, organs, or cells.  After TCDD enters
the host  from an  environmental  source, it is distributed to target organs  or storage
depots.   However, TCDD can be mobilized from depots and redistributed to target
organs or other  depots, producing an in vivo bioavailability that should be recognized.
Although there are no data on the mobilization of TCDD in humans, knowledge of the
bioavailability of TCDD by mobilization has been documented in other animals.  This
aspect of bioavailability is extremely important  because any TCDD stored in the
human body in the adipose or other tissue is always  a source that could be mobilized
under a number of conditions and has the potential  to  produce toxicity in the  host.
The  determination of the end points of toxicity and quantification  of the degree  of
toxicity will be dependent on the internal distribution  of TCDD.

     It  is important to know  how  host  factors such as possible pregnancy,  age,
nutritional status, and prior  exposure  influence  the  in   vivo  bioavailability  by
mobilization and redistribution of TCDD, with the possibility of altered end points  or
                                       38

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altered degree of toxicity.  Moreover, several critical organs and cellular targets are
likely, so that all of the implications of redistribution are not known.  Nevertheless, it
is important to enhance excretion, directly or by redistribution, to limit distribution or
redistribution to such sensitive tissues as fetuses and gonads, and to  prevent secondary
exposure via breast milk.

3.3.2  Research Needs

      There are very few data on  animals or humans in  which the concentration of
TCDD from an in vivo bioavailable source is known.  Studies and techniques are needed
for the  determination  of TCDD from a primary exposure and from  a secondary
exposure through mobilization and redistribution.

      The determination of the TCDD body burden  in humans is needed,  with adipose
tissue being the most important  tissue to be examined.  This  would permit a  direct
comparison to the other halogenated hydrocarbons  and dioxin and furan  isomers that
would provide a background profile of  the extent of  contamination of these compounds
in the human population.

      Additional studies should determine the  residue  of TCDD in other organs of
humans that might determine TCDD target organs as well as  the possible mobilization
and redistribution of TCDD.  This would also suggest the possible metabolic pathways
and/or secretion or excretion patterns in humans.

      To  perform many  of the studies, there is a need to  develop and validate  assays
for TCDD that are rapid and economical, either in vivo or in vitro, and that can be
used to determine the concentration  of the  TCDD  in the various organs of  the body.
This would be necessary for  studies to determine  mobilization and redistribution of
TCDD.

      Analytical chemical  methods for the direct measurement of TCDD are available
for fat and for breast  milk  analyses (see Table 1.1  in Chapter 1).  These  methods
should be applicable  to other organs, some of which  can  be assayed by biopsy.  Biopsies
would permit the determination of TCDD  in various organs which could be related to
the microscopic examination of the biopsy.

      Toxicity end points  could be  used to estimate the bioavailability of TCDD, but
they lack specificity.  It is well established that TCDD produces chloracne in humans
and some animals. Although this response is not specific for TCDD, the assay could be
a  sensitive indicator   of TCDD.   Enzyme  changes  and   alteration  of  porphyrin
metabolism could be explored as indicators of TCDD bioavailability, although they also
lack  specificity.    Extracts  of  various  matrices  could   be  assayed  for  TCDD
bioavailability by indirect in vitro  methods, such as induction of rat hepatocyte aryl
hydrocarbon  hydroxylase  (AHH)  activity, human  lymphocyte AHH activity,  and
porphyrin changes in human f ibroblasts, although there are no data on the alteration of
porphyrin metabolism in humans as  seen in other animals.  These indirect methods are
not specific for TCDD, and  it should  be recognized that other dioxins and furans can,
and do, affect these end points, confounding the results.  Specificity might be  obtained
by methods that are under  development for estimating the concentration  of  TCDD
using radio!mmunoassay and monoclonal antibody techniques.


                                      39

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                    3.4 Host Factors Influencing Bioavailability

      Many host factors can modulate the bioavailability of TCDD with an increase or
decease of availability. Because there are many host factors, only some of the well
known factors will be addressed.

3.4.1 Dietary Factors

      Dietary consumption  patterns  influence  exposure and   may  also  influence
bioavailability by affecting absorption and rate  of retention in the  gastrointestinal
tract. The nutritional status of the host in  terms of deficiencies, malnutrition, and
specific diets (e.g., high or low fat)  will influence  bioavailability by increased or
decreased  uptake.   Body fat content will have a major effect on distribution and
mobilization.

3.4.2 Genetic Differences

      It  is  well  known  that  genetic  differences   can   influence  absorption,
biotransformation,  receptor interaction, and  excretion of  many toxic  agents in
numerous species.   However, very little is known about these factors in humans.
Although few data are available, it  is recognized that genetic differences could be a
major factor determining the variability of bioavailability in humans and subsequent
toxic manifestations.  Studies should be undertaken with responsive and nonresponsive
mice to indicate a possible range of effects.

3.4.3 Age

      Both behavioral  and physiological characteristics  related  to  age  influence
bioavailability.  Factors such as dermal  penetrability and gastrointestinal absorption
change with age.   Also, there  is evidence from  animal studies for  oncogenetic
development of cellular receptors for TCDD.

3.4.4 Concomitant Exposures

      TCDD  is similar to  other chlorinated hydrocarbons in  its  manifestation of
toxicity.  Concurrent  exposures  to other chlorinated  hydrocarbons as well  as furans
and  other dioxin  isomers may influence the bioavailability of TCDD by competition,
inhibition,  or enhancement.  However,  very  few data are available on this topic
relating to humans.

3.4.5 Exposure History and Other Factors

      Prior exposure  to organic chemicals may  enhance or reduce the  uptake  and
retention of TCDD, thus influencing the bioavailability.

      Other  host factors to be  considered  are behavior, lifestyle, and  health  and
disease states. Data are insufficient to warrant  high priority consideration of theie
factors.  However, information  from occupational studies may assist  in reevaluating
the importance of these factors.

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3.4.6 Research Needs

      Adequate consideration of research needs in the area of host factors affecting
bioavailability is hampered by a lack of knowledge regarding critical target organs and
the  best  choice  of an animal model  for  human  toxicity.  The  appropriate  animal
species to  use  as a  model  for studying  host  factors, tissue distribution,  and
mobilization from body stores is needed because data show that the tissue distribution
of TCDD differs in monkeys and rodents (see discussion  in Section 3.5).  Many  end
points of toxicity could be studied; however, the critical end point in TCDD toxicity is
not known.  Some end points that should be considered are:  elevation  of blood lipids,
increased   production   of  porphyrins,   impaired   immune  response,  peripheral
neuropathies, and dermal  disorders such as chloracne.   Other  biochemical markers
need to be determined, especially those specific to TCDD; however, these would also
most likely  be indicative of the other dioxins and f urans.  Additionally, the extent to
which data on other dioxin and fur an isomers can  be applied to TCDD should be
evaluated.

               3.3 Ihterspecies Differences Affecting Bioavailability

      Data derived  directly from in vivo studies of  TCDD bioavailability in  humans
would obviously be  most useful in addressing the issue of bioavailability. However, for
ethical and other reasons, it is likely that most of the data on bioavailability will, of
necessity, be derived from in vivo studies using laboratory  animals as surrogate testing
species.

      Therefore, it was deemed imperative that these anticipated animal studies on
bioavailability be planned, conducted,  interpreted, and extrapolated to humans in a
scientifically  appropriate  manner that  accommodates all the  information  on  the
interspecies differences affecting bioavailability.  These differences can be delineated
into two groups:  the more  general conceptual interspecies differences applicable to
many  xenobiotics,  and  the  more  specific  interspecies  differences  of  particular
importance  in evaluating the bioavailabiliity  of TCDD and other halogenated dioxins
and f urans.

      The interspecies  differences affecting bioavailability  by  the three principal
routes of exposure (dermal, ingestion, and  inhalation) would include, but  are  not
limited to, the following variables among humans and the various laboratory animal
species likely to be  used in the in vivo studies of bioavailability.

3.5.1 Dermal Route

      The dermal   route  of  exposure  to TCDD provides a  direct  mechanism  for
absorption  of  the  toxicant.   This exposure can result  from  direct  contact with
contaminated soil or through air transport of dust or fly ash.

      3.5.1.1  Dermal Studies

      Anatomical and physiological differences  among humans and  various laboratory
animal  species would  include  factors  such  as  the presence  or absence   of   an

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integumentary hair coat, the thickness of the hair coat, the thickness of the epidermis
and dermis, the presence  or absence of integumentary  adnexai structures  such as
sebaceous and  sweat glands, the  comparative aspects of the  subcutaneous fat and
vasculature  and lymphatics, the  comparative ratios  of  skin surface area  to  body
weight, the  comparative enzymatic capabilities  of the dermis, and  the comparative
potential for binding to plasma proteins after dermal absorption.

     3.5.1.2 Research Needs

     Evaluation of the  suitability of  various in vivo and in vitro models is needed for
predicting dermal absorption of TCDD.  Dermal  uptake of TCDD should be examined
in several species  in vivo.   Direct absorption  from various matrices, such as soil and
fly ash, should be examined. Species that  might  be considered  include pigs (neonatal
skin), rats,  guinea pigs, and mice, including dermal  uptake by both responsive and
nonresponsive strains of mice.  Uptake by various species in  vivo  should be compared
to transport in at least one in vitro model using skin explants from the same species as
well as uptake in human skin culture.   Species differences in  uptake in vitro should be
compared with species differences in vivo  to determine whether the  in vitro  model is
predictive of the in vivo data and to determine  in which species dermal absorption
most closely resembles absorption by human skin.

3.5.2 Oral Route (Digestion)

     In a second type of exposure, ingestion, bioavailability is a  function of  whether
TCDD  intake  is  from  eating  contaminated  foods  or  from  exposure  to TCDD-
contaminated soil  or dust.  If TCDD-contaminated food  is consumed, bioavailability
becomes analogous to bioaccumulation.

     3.5.2.1 Oral  Studies

     Comparative studies  of bioavailability using various laboratory animal species as
surrogates for humans should consider the  following variables:   interspecies dietary
patterns (herbivorous, carnivorous, or omnivorous), anatomical differences such as the
presence or absence of cecum, comparative transit times via  the gut, comparative
aspects of  biliary  salt production and  pancreatic  enzyme  production, comparative
differences  in pH within the gut, comparative aspects of enterohepatic circulation and
gut flora, and intestional vasculature and lymphatics.

     3.5.2.2 Research Needs

     In  bioavailability  studies of  TCDD, feeding  contaminated  soils  to various
laboratory animals  has produced substantial  differences  in response.   Thus, the
differences  in bioavailability that have been identified warrant further studies.   Some
of these further studies should use variations of the  more conventional in vivo test
procedures that have been utilized in bioaccumulation studies with certain laboratory
species such a guinea pigs and rats.

     Other  types  of studies that  should be done, the results of  which could lead to
generalizations  that  can  be modeled,  include  in  vitro  gut  sack  and  substituted

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gastrointestinal content  studies  as  a means to  estimate release of  TCDD  from
matrices.  Gut sack studies allow the investigator to examine the role of different
areas of  the intestinal tract in absorption and  transport.   This model is easily
adaptable to several animal species, including the human digestive tract from surgical
specimens.   The  human specimens  would  be  necessary to extrapolate data  from
animals to humans.

      Substituted gastrointestinal content studies are currently carried out by the U.S.
Food and Drug Administration (FDA)  and the  U.S. Department  of Agriculture to
determine the release of disiodgeable residue of pesticides and the release of active
ingredients from medications,  particularly generic equivalents.  This approach could be
coupled  with high-resolution  analytical  techniques  to estimate the  effect  of the
digestive process (time, pH, and volume) on dislodging TCDD from matrices.

      The monitoring of the human diet by the  FDA should be sufficient to estimate
the bioavailability of TCDD from diet.  However, special situations  may need direct
study, such as people on a high-fat diet, people growing gardens next to dump sites, or
children ingesting dirt.

3.5.3 tihalation Route

      Municipal waste incinerators have been well characterized as  being potentially
chronic, dispersive sources of low  levels (parts per trillion) of  chlorinated dioxin
isomers into the atmosphere.  At some time following release from incinerator stacks,
those isomers not already adsorbed or absorbed to  participates will become bound to
participates as the compounds reach  sublimation.  Therefore, human exposure can be
predicted to occur by  inhaling TCDD-bound  participates into the respiratory tract.
Bioavailability would then become a  function of  particle size, the chemical nature of
the participate, the distribution of TCDD in relation to particle size, the dynamics of
particle-size retention in the lungs, and the action of the lungs in dissolving, digesting,
or extracting TCDD from the particles.

      3.5.3.1 Inhalation Studies

      Inhalation of TCDD from environmental matrices should be studied in at least a
rodent model.  One such study should determine the bioavailability of TCDD in rats
exposed to dusts generated from  specific sites  or  matrices.  The matrices  should be
selected to represent extreme conditions of absorption, so that models can be built to
assess human risk without testing every contaminated site or matrix.

      The dusts (participates) should be generated by a dust feeder that allows particle
sizing, such as the Wright Dust Feeder.   The dust should be analyzed prior to animal
exposure. The analysis should include  the  compounds  present,  the type of matrix, and
the composition of the matrix. After nose-only exposure, the biochemical markers and
toxicity  indicators should be  determined.  A  major question to  address  is which
compounds are retained compared to  compounds in  the original sample.  The nose-only
inhalation procedure eliminates the possibilities of dermal deposition and absorption,
as well as oral ingestion from grooming.

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     3.5.3.2 Research Needs

     In  view  of  the  predominant  concern  regarding  exposure  to  particulates,
anatomical and physiological differences should be addressed.  Anatomical differences
among humans and the various laboratory animal species should be examined in regard
to the upper respiratory tract,  the  bronchiolar tree, the  alveoli,  and the pulmonary
vasculature and lymphatics.

     Physiological differences  would include respiratory volume,  respiratory rate,
participate  sedimentation   rates  for  the   upper  and  lower  respiratory   tracts,
comparative aspects of clearance via mucociliary apparatus, retention time, and the
ultimate fate for particulates that do reach the alveoli.

     When  designing studies of  bioavailability in laboratory animals, consideration of
these  general issues should  be supplemented by  more  specific  issues  of  special
importance for the chlorinated  dioxins and furans that are discussed in the following
two sections.

            3.6  Pharmacokinetics And Structure-Activity Relationships

     Absorption, distribution, and excretion of dioxins and furans have been studied in
a number of species. These compounds were found principally in the liver and fat of
all species examined. Differences among species in metabolism or distribution do not
appear to account for differences in  species sensitivity.

3.6.1  Fharmaookinetic Studies

     The most toxic isomer of dioxins and furans is substituted in all four  lateral
2, 3, 7, and  8 positions; however, extensive in vitro and in  vivo quantitative structure-
activity relationships have not been  fully developed. This information could readily be
used to predict the toxic end points  and human toxicity of isomers of this  large class
of toxic  environmental  agents.  The few structure-activity  relationships of dioxin
isomers indicate that TCOD  is the most  toxic compound, with the other isomers being
less toxic.  Estimating the toxicity  of mixtures of these various dioxins and furans is
not possible, simply because too little is  known about the toxicity  of  the individual
compounds or the possible interaction of the compounds.

3.6.2  Research Needs

     The areas that should be studied include:

     •  the effects of structure on the biologic and  toxic activities of dioxins  and
         furans;

     •  the development and validation of quantitative in vitro bioassays that can be
         used to predict in vivo effects of dioxins and furans;

     •  a study of the  in  vivo  interactive effects  of dioxins and furans and  the
         determination of additive, synergistic, and antagonistic effects;

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     •   a study  of dioxin and  fur an isomers  and the  effects  of structure on
         pharmacokinetics in exposed animals;

     •   the role of interactive effects on the pharmacodynamics of mixtures; and

     •   the  effects of receptor level modulation (e.g.,  by PCBs and PBBs) on the
         toxicity of individual dioxins, f urans, and their mixtures.

                               3.7 Epidemiology

     When reviewing the results of animals studies, it  is  easy to look at only one
compound, TCDD.  However, when reviewing epidemiological data, it is extremely
difficult to deal with only one compound, because most  exposure groups have been
exposed to a  mixture of compounds or isomers. For this reason, we feel that the focus
of dioxin research must  address a number  of compounds, including dibenzodioxin,
dibenzofuran isomers, and halogenated hydrocarbons.

3.7.1  Epidemiological Studies

     There  have  been  a  large number of animal studies  on the adverse biological
effects  of  TCDD exposure such  as  hepatotoxicity,  porphyria, dermal changes,
teratogenicity   (e.g., cleft palate and  hydronephorosis),  and functional toxicity,
including  modulation of immune response.   Long-term  effects of  exposure  are of
interest, especially  as  exposure may  impact humans.  Animal studies have  shown
relatively long-term immunosuppression following perinatal exposure and increased
incidence of soft-tissue neoplastic disease.    Despite the  abundance of  data from
animal studies, a great need remains for studies of the effects of TCDD exposure in
humans.

     Regarding the carcinogenicity of TCDD, several epidemiological studies have
found a  significant  positive association  between exposure to dioxin-contaminated
phenoxy  acids  and/or  chlorophenols  and soft-tissue  sarcomas  and  non-Hodgkins
lymphomas.    Additionally,  in  several  small  industrial cohorts exposed  to TCDD-
contaminated  chemicals,  several  cases of soft-tissue sarcoma have appeared  where
none were expected because of the relative rarity of the disease.  Also, chloracne was
either suspected or confirmed.  These studies  have several limitations that preclude
the estabishrnent of a causal relationship with  TCDD exposure at  this time, although
several   other   studies  showing   no   effects  suffer  from  limitations  as   well.
Epidemiological studies are  needed to  address  the toxic manifestations of TCDD in
humans.

     The results of immune function studies in humans following TCDD (or related
chemicals) exposure  are ambiguous. Children  exposed in the Seveso accident were
shown  to  have  significantly  elevated lymphoproliferative  responses,   but  other
published  studies  have  reported  no effects.  Unpublished studies  have  indicated
changes  in lymphoproliferative responses  (both  enhancement and  suppression) and
shifts in lymphocyte  subpopulations.   Studies  of  individuals  exposed  to  PCBs have
shown that PCBs  are immunomodulators in humans, causing suppression  of delayed
hypersensitivity reactions,   enhancement  of  lymphoproliferative  responses,  and

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increased incidence of infectious  diseases.  It is not known if these end points  can
occur in humans exposed to TCDD.

3.7.2  Research Needs

      Following is a list of research needs related to human health:

      •   Methods need to be developed to identify persons who have been exposed to
         TCDD and  related  chemicals, and to identify  more precisely  the  actual
         compounds  to  which they were exposed.  This will  enable researchers to
         evaluate more fully the human health effects of exposure.

      •   Because  of the potential   for  interaction  of  the  halogenated  cyclic
         hydrocarbons  (additive, syngeristic, or  antagonistic  effects),  studies  are
         needed to investigate  the  effects  of  multiple  compounds.  This includes
         identifying human populations exposed to a mixture of compounds, describing
         the  effects  of  exposure in  these  populations,  and  performing laboratory
         studies on the interaction of compounds.

      •   When possible, studies should be performed using in vitro exposure of human
         tissues to dioxins (i.e., human tissues in  culture)  and the  results  of these
         studies utilized to evaluate the studies of in vivo exposure of animals and to
         facilitate extrapolation to humans.

      •   Additional epidemiological studies should be accomplished with cohorts  not
         exposed to TCDD or similar  chemicals, to establish the baseline  for humans
         for the anticipated end points of toxicity. Although data from animal  studies
         are available in this area, data for human exposure are needed.

      •   Based on  the sensitivity of the immune system (animal studies) to  chemical
         exposure  and based on preliminary findings in human populations exposed to
         TCDD about the  importance of the immune system in resistance to neoplastic
         and  infectious disease,  additional studies are needed to describe more fully
         the  effects  of  TCDD  and  related  chemicals on the  immune function in
         humans.  Establishing the proper measure of immune function may enable an
         indication of low-level  exposure in humans.

                         3.8 Need For Supply Of TCDD

      To  date, the toxicity of halogenated dioxins has been determined with  studies
using only the single compound, TCDD. To evaluate and understand the problems with
the complex environmental matrices containing many halogenated dioxins  and  f urans,
mixtures of dioxins with and without  f urans should be studied.  This, in turn, requires
sufficient quantities of these compounds for toxicology studies.

      EPA maintains a repository of purified, analytical-grade pesticides and industrial
chemicals for  use by scientists throughout the  world.   This is a very successf Jl
program  supplying small  quantities  of standards for chemical  assays.   In general,
commercial chemical companies do not make  dioxins and f urans available  to

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toxicologists because of a limited market. Supplies of these chemicals are needed in
sufficient quantities and purities for use by toxicologists.

      By expanding current and proposed studies to include other  dioxins and furans
with the study of TCDD, data will be generated that will be significantly more useful
to EPA by better focusing on the problems in dump sites and other areas of interest.
If the compounds are not available for a few years, answers to the critical questions
about dump sites could be delayed a decade or more because many of the studies would
have to be repeated to incorporate the other  dioxins and furans.  Thus, we recommend
that the EPA support production and supply these compounds to investigators studying
dioxins and furans.

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Donald Barnes



Oavid Bayliss



Judith Bellin



Diane Courtney



Sergio Fabro



Robert Faith



Marilyn Fingerhut



Michael Gallo



Joyce Goldstein



Maurene Hatch



Greg Kew



Renate Kimbrough



Richard Kociba
Bioavailability to Humans Working Grotj>



           PARTICIPANTS



                            Barry Korb



                            David Marlow



                            James Melius



                            Patrick O'Keefe



                            Dhun Patel



                            Stephen Safe



                            John Schaum



                            Arnold Schecter



                            Barclay Shepard



                            Ellen Silbergeld



                            Frode Ulvedal

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                                   CHAPTER*

                                   SUMMARY
      In order to understand the effects of transport and fate  of dioxins and related
isomers through environmental media and, ultimately, bioavailability to humans, there
are four areas of research needing critical attention:  physical and chemical data, field
studies, modelling, and human effects  studies.  Implicit in all of these is a need for
adequate supplies of and laboratory standards for all possible isomers and compounds
of dioxins and furans, to ensure quality assurance and quality control.

                         4.1 Physical and Chemical Data

      The physical and chemical properties of TCDD isomers, other dioxins, and furans
need to be further investigated in order to analyze their behavior within environmental
media.  This workshop has recommended the use of the structure-activity relationship
approach, based on thermodynamic consideration. An increased data base will further
enhance estimations of partitioning behavior across abiotic interfaces, which will aid
estimates  of bulk  transport  and  intermedia  transfer  processes.    Understanding
intermedia  transfer processes can  aid in the  investigations  of methodologies  for
degrading dioxins in the environment, through photolysis, chemical transformations, or
biological processes, ultimately, expansion of the information on physical and chemical
properties  of dioxins  and related isomers will assist the  investigations of ecosystem-
level  processes affecting and being  affected by these chemicals and  will add further
information to answer questions about bioavailability to humans and effects on  human
tissues and organs.

                                4.2 Field Studies

      In order to supplement and enhance the results of laboratory  research on basic
questions  of physical and chemical behavior of dioxins, output from  field studies is
needed. Information can be used to verify and validate modelling and laboratory work,
as well as  provide basic data on observed environmental effects.  Particularly for the
issue of bioavailability in ecosystems, a full-scale field study needs to be conducted  at
a highly contaminated site.  This should include studies of transport and fate, chronic
effects, and ecological processes.  Such  a field study  should be  closely linked  to
laboratory investigations.

                                  4.3  Modelling

      Further efforts need  to be  made  to develop more accurate  and applicable
mathematical models to predict exposure  concentrations  of dioxins  in environmental
media. Concentrations of dioxins over time in organisms as a function of dose need  to

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be measured for  model development.  The food-web conceptual model  presented  in
Chapter 2 can be a base upon which calculations can be made giving initial estimates
of dioxin concentrations for components.  This generic model can be applicable to both
terrestrial  and aquatic ecosystems,  with  more specific detail  added  as  needed.
Development of ecosystem-level  models should be  one major  goal of the full-scale
field study program.

                                 4.5 Summary

     Running as a common thread throughout this report is the  need for further basic
research on  the  physical  and chemical  makeup  of  dioxins  and related  isomers,
compounds,  and  mixtures.    But even  more   relevant  to  the  end  point of  all
environmental regulations,  effects on humans, is a  need for a basic understanding of
how dioxins relate to the environmental media into which they are released. How they
affect  and are affected by the physical and chemical processes  with which they come
in contact  as they are transported through the biosphere speaks to  the  ultimate
questions of control and degradation, of exposure, bioavailability, and risk. A focused
research plan would seek to understand  these most basic processes and  interactions,
from which  base of information  adequate assessment  can be  made of the levels of
regulation most practicable.
                                      50

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                            APPENDIX
                          PARTICIPANTS
William Adams
Monsanto (NIB)
800 N. Lindbergh
St. Louis, MO 63167

Donald Barnes
U.S. Environmental Protection
 Agency (TS-788)
401 M Street, SW
Washington, DC 20*60

David Bayliss
Carcinogen Assessment Group
U.S. Environmental Protection
 Agency (RD-689)
401 M Street, SW
Washington, DC 20460

Judith Bellin
WH-562B, Room 5242
U.S. Environmental Protection
 Agency
401 M Street, SW
Washington, DC 20460

3immy Boyd
J.M. Huber Corporation
P.O. Box 2831
Borger, TX 79008

Erich Bretthauer, Director
Off. Environ. Proc. & Effects Res.
U.S. Environmental Protection
 Agency (RD-682)
401 M Street, SW
Washington, DC 20460
Brian Butterworth
Environmental Research
 Laboratory
U.S. Environmental Protection
 Agency
6201 Congdon Blvd.
Duluth, MN  55804

Peter Chapman
University of Minnesota
Dept. of Biochemistry
140Gortner Lab
1479 Gortner Avenue
St. Paul, MN 55108

Morris Chelsky
Diamond Shamrock Tower
717 N. Harwood
Dallas, TX  75201

David Cleverly
Off. of Air Quality Planning and
 Standards
U.S Environmental Protection
 Agency (MD-12)
Research Triangle Park, NC 27711

Michael Cook
WH-562
U.S. Environmental Protection
 Agency
401 M Street, SW
Washington, DC  20460

Philip Cook
Environmental Research
 Laboratory
U.S. Environmental Protection
 Agency
6201 Congdon Blvd.
Duluth, MN 55804
                                 51

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Diane Courtney
Health Effects Research Laboratory
U.S. Environmental Protection
  Agency (MD-82)
Research Triangle Park, NC  27711

Russ Cristman
Dept. of Environ. Sciences &
  Engrg. School of Public Health,
Duluth, MN 55804
University of North Carolina
Chapel Hill, NC  27514

Donald G. Crosby
Environmental Toxicology
  Department
University of California-Davis
Davis, CA  95616

Ron Dagani
Chemical & Engineering News
1155 16th Street, NW
Washington, DC  20036

Michael Dellarco
OMSQA (RD-680)
U.S. Environmental Protection
  Agency
401 M Street,  SW
Washington, DC  20460

Paul DesRosiers
ORD-OEET
U.S. Environmental Protection
  Agency (RD-681)
401 M Street,  SW
Washington, DC  20460

Robert Diaz
Virginia Institute of Marine
  Sciences
Glouster Point, VA 23062

Thomas Duke
Environmental Research
  Laboratory
U.S. Environmental Protection
  Agency
Sabine Island
Gulf Breeze, FL  32561
Aubry Dupuy
U.S. Environmental Protection
  Agency
Building 1105
NSTL, MS  39529

Tom Evans
Off. of Waste Enforcement
U.S. Environmental Protection
  Agency (WH-527)
401 M Street, SW
Washington, DC  20460

Sergio Fabro
Columbia Hospital for Women
2425 L St., NW
Washington, DC  20037

Robert Faith
Animal Care Operations
University of Houston
4800 Calhoun
Houston, TX 77004

Walter 3. Farmer
Dept. of Soil and Environ. Science
University of California-Riverside
Riverside, CA  92521

Marilyn Fingerhut
NIOSH
4676 Columbia Parkway
Cincinnati, OH  45226

Virgil Freed
Dept. of Agricultural Chemistry
Oregon State University
Corvallis, OR 97331

Randy A. Freeman
Monsanto
800 N. Lindbergh
St. Louis, MO 63137

George F. Fries
Pesticide Degradation Laboratory
Beltsville Agricultural Res. Center
Beltsville, MD  20705
                                 52

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Michael Gallo
UMDN3
Rutgers Medical School
P.O. Box 101
Piscataway, NJ 08854

Bernard Goldstein
Assistant Administrator
Office of Research and Development
U.S. Environmental Protection
  Agency
401 M  Street, SW
Washington, DC 20460

Joyce Goldstein
NIEHS
P.O. Box 12233
Research Triangle Park, NC  27709

Rizwanul Haque
U.S. Environmental Protection
  Agency
401 M  Street, SW
Washington, DC 20460

Robert Harless
Environmental Monitoring Systems
  Lab.
U.S. Environmental Protection
  Agency (MD-67)
Research Triangle Park, NC  27711

Mark Harwell
Ecosystems Research Center
Corson Hall
Cornell University
Ithaca, NY 14853

Maurene C. Hatch
Division of Epidemiology
Columbia University
600 W. 168th St.
New York, NY 10032

Ralph Hazel
324 E.  llth St.
Kansas City, MO 64106
Robert J. Huggett
Virginia Institute of Marine Science
Gloucester Point, VA 23062

Wayne Ivie
USDA, VTERL
P.O. Drawer GE
College Station, TX 77841

Darcy Johnson
Environmental Research
  Laboratory
U.S. Environmental Protection
  Agency
6201 Congdon Blvd.
Duluth, MN  55804

Kenneth L. Johnson
Environmental Research
  Laboratory
U.S. Environmental Protection
  Agency
6201 Congdon Blvd.
Duluth, MN  55804

Steve Kennel
Oak Ridge National Laboratory
P.O. Box Y, Biology Division
Building 9220
Oak Ridge, TN  37830

Greg Kew
Office of Health &  Env. Assessment
U.S. Environmental Protection
  Agency (RD-689)
401 M Street, SW
Washington, DC 20460

Renate Kimbrough
Centers for Disease Control
1600 Clifton Road
Atlanta, GA 30333

Richard J. Kociba
Dow Chemical Company
Midland, MI  48640
                                53

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Takeru Higuchi
2201 W. 21st St.
Lawrence, KS 660**
Christopher Kouts
U.S. Dept. of Energy
760* Jervis Street
Springfield, VA  22151

Tim Kubiak
U.S. Fish and Wildlife Service
University of Wisconsin-Green Bay
Green Bay,WI  5*302

Douglas Kuehl
U.S. Environmental Protection
 Agency
ERL-Duluth
6201 Congdon Blvd.
Duluth, MN  5580*

Bill Lamason
Air Management Technology
 Branch
OAQPS
U.S. Environmental Protection
 Agency (MD-1*)
Research Triangle Park, NC 27711

Ray R. Lassiter
Environmental Systems Branch
Athens Environ. Res. Lab.
College Station  Road
Athens, GA  30613

John C. Loper
Dept. of Microbiology and
 Molecular Genetics (ML-52*)
College of Medicine
University of Cincinnati
Cincinnati, OH  45267

David Marlow
NIOSH
4676 Columbia Parkway
Cincinnati, OH  45226
Barry R. Korb
OSWER (WH562-A)
U.S. Environmental Protection
 Agency
401 M Street, SW
Washington, DC 20*60

John McCarthy
Oak Ridge National Laboratory
P.O. Box X
Oak Ridge, TN 37831

Danny D. McDaniel
U.S. Environmental Protection
 Agency
Building 1105
NSTL, MS 39529

James Craig McFarlane
Environmental Research
 Laboratory
U.S. Environmental Protection
 Agency
200 SW 35th St.
Corvallis, OR 97333

Jay Means
Chesapeake Biological Laboratories
P.O. Box 38
Solomons, MD  20688

James Melius
Robert A. Taft Laboratories
*676 Columbia Parkway
Cincinnati, OH *5226

 Theodore Mill
 PS 273
 SRI
 Menlo Park, CA 9*025

Ronald K. Mitchum, Dir., QA Div.
Environmental Monitoring Systems
 Lab.
U.S. Environmental Protection
 Agency
P.O. Box 15027
Las Vegas, NV  8911*
                                5*

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Leland Marple
Syntex Research
3401 Hillview Ave.
Palo Alto, CA

Arthur Martell
Dept. of Chemistry
Texas A & M University
College Station, TX  77843

3ay Murray
Syntex Research
3401 Hillview Ave.
Palo Alto, CA 94304

Patrick O'Keef e
NY State Department of Health
Center for Laboratories and
  Research
Empire  State Plaza
Albany, NY  12201

Dhun Patel
N3 Dept. of  Environmental
  Protection
Off. of Science and Research
190 W. State St.
Trenton, N3  08625

Dennis Paustenbach
Syntex Corporation-4A
3401 Hillview Ave.
Palo Alto, CA  94303

Warren  Piver
MD-A2-05
NIEHS
P.O. Box 12233
Research Triangle Park, NC 27709

Hap Pritchard
Environmental Research
  Laboratory
U.S. Environmental Protection
  Agency
Sabine Island
Gulf Breeze, FL  32561
Charles Morgan
Off. of Waste Enforcement
U.S. Environmental Protection
  Agency (WH-527)
401 M Street, SW
Washington, DC  20460

Michael Roulier
Municipal Environmental Research
  Laboratory
26 West St. Clair Street
Cincinnati, OH 45268

Thomas Sabourin
Battelle Columbus Labs
505 King Ave.
Cincinnati, OH 43201

Stephen Safe
Texas A&M University
College of Veterinary Medicine
Dept. Vet Physiology &
  Pharmacology
College Station,  TX  77843

Walter M. Sanders III
Environmental Research
  Laboratory
U.S. Environmental Protection
  Agency
College Station Road
Athens, GA  30613

John Schaum
U.S. Environmental Protection
  Agency (RD-689)
401 M Street, SW
Washington, DC  20460

Arnold Schecter
Upstate Medical  Center
College of Medicine
Clinical Campus at Binghamton
Binghamton, NY  13901
                                55

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 John F. Quenson
 Pesticide Research Center
 Michigan State University
 East Lansing, MI  4882*

 Gerald Rausa
 Off. of Research and
  Development/RSS
 U.S. Environmental Protection
  Agency
 *01 M Street, SW
 Washington, DC 20460

 Barclay Shepard, VA Central Off.
 Agent Orange Project Off. (IDA?)
 Suite 308, Shoreham
 810 Vermont Ave., NW
 Washington, DC 20*20

 Ellen Siibergeld
 Environmental  Defense Fund
 1525 18th Street, NW
 Washington, DC 20036

 Ronald A. Stanley
 4119 Military Road, NW
 Washington, DC 20015

 HQ AFESC/RDVW
 Attn:  Capt. Terry Stoddart
 Tyndall AFB, FL  32*03

 Richard E. Tucker
 Dynamac Corporation
 lll*ORockville Pike
 Rockvilie, MD  20852

 Frode Ulvedal
 Office of Health Research
 U.S. Environmental Protection
  Agency
 *01 M Street, SW
 Washington, DC 20*60
Robert Schreiber
Dept. of Natural Resources
DEQ Administration
P.O. Box 1368
Jefferson City, MO  65102

Jerry Schroy
Monsanto
800 N. Lindbergh
St. Louis, MO  63167
Barbara T. Walton
Environmental Sciences Division
Oak Ridge National Laboratory
P.O. Box X
Oak Ridge, TN  37831

John E. Wilkinson
23*0 Taylor Way
Tacoma, WA  98*01

Armon Yanders
Environmental Trace Substances
Route 3
Columbia, MO 65203

Alvin Young
Office of Science & Technology
 Policy
New Executive Office Bldg.
Room 5005
Washington, DC  20506
                                 56
U.S.Government Print,ng O'fice: 1986 - 646-014/4O004

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