TRIBUTYLTIN SUPPORT DOCUMENT
       Office of Pesticide Programs
Office of Pesticides and Toxic Substances
   U.S. Environmental Protection Agency
            401 M Street, S.W.
          Washington, DC  20460
              December 1985

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                       Executive Summary
       This Tributyltin Support Document presents the basis for
 the initiation of a Special Review of all pesticide products
 containing tributyltin (TBT) active ingredients used as paint
 additives (antifoulants) to inhibit the growth of certain aquatic
 organisms.  The TBT paints are primarily applied to boat and ship
 hulls.   These TBT compounds include:   bis-(tributyltin) oxide,
 bis(tributyltin) adipate,  bis(tributyltin)  dodecenyl succinate,
 bis(tributyltin) sulfide,  tributyltin acetate, tributyltin acrylate,
 tributyltin fluoride,  tributyltin methacrylate, and tributyltin
 resinate.

      The initiation of the Special Review  is  based on  the Agency's
 determination that the use of  the TBT compounds in antifoulant
 paints  may result in TBT exposure to  nontarget aquatic  organisms
 at  concentrations resulting in acute  and chronic effects.   The
 Agency  has determined  that the risk criteria,  as described in 40
 CFR 162.11 are met or  exceeded by use of these TBT antifoulant
 paints.

      The Agency evaluated bioassay studies  which indicated that
 the TBT  compounds are  highly toxic, frequently at the parts per
 trillion (ppt)  level,  to nontarget marine and  freshwater aquatic
 organisms.  Toxicity tests have  found adverse  affects to molluscs
 at  levels  of  60  and  100 ppt.   The laboratory toxicity measure
 which the  Agency  used  to evaluate the acute  hazard was  the median
 lethal concentration  (LCso)  which kills  50 percent of the  test
 organisms  after  a designated time period.  The Agency also reviewed
 numerous  aquatic  toxicity  studies which  indicate that the  TBT
.compounds  cause  chronic hazards  such  as  anatomical abnormalities,
 growth effects, and is bioaccumulated.

      Environmental monitoring data measuring  TBT concentrations
 in the Great  Lakes and U.S. coastal waters were  compared to the
concentrations identified  as causing  adverse effects  in the
laboratory toxicity studies.  From this  comparison,  the Agency
has determined that the risk criteria, as described  in  40  CFR
162.11 are met or exceeded by use  of  these TBT  antifoulant paints.

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                         ACKNOWLEDGEMENTS
       Janet  L. Anderson
       Kyle R. Barbehenn
       Michael A. Champ
       Stanley A. Cook
       John D. Doherty
       Robert K. Hitch
       Peter Kuch
       Thomas W. Purcell
       Allan J. Reiter
       Michael Rexrode
       Philip J. Ross
       Myra Smith
       Linda K. Vlier
       Suzanne Wells
Project Team

  Plant Pahtologist, OPP/BUD
  Biologist, OPP/HED
  Senior Science Advisor,  OPPE
  Assist. Review Manager,  OPP/RD
  Toxicologist,  OPP/HED
  Ecologist, OPP/HED
  Supervisory Economist, OPP/BUD
  Environmental  Scientist,  OWRS
  Chemist, OPP/HED
  Fisheries Biologist,  OPP/HED
  Attorney, OGC
  Microbiologist,  OPP/BUD
  Senior Review  Manager, OPP/RD
  Environ. Protection Specialist, OPPE
Additional assistance also provided by:

       Robert J. Huggett     '•
       Virginia Institute of Marine Science
       College of William and Mary
       Gloucester Point, Virginia

       Peter F. Seligman
       Marine Environmental Branch
       U.S. Naval Ocean Systems Center
       San Diego, California

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                      TABLE OF CONTENTS
 I.     INTRODUCTION  	   1-1

       A.  General Background and Organization  	   1-1
       B.  Legal Background  	„	   1-1
       C.  Chemical  Background	   1-2

 II.    ASSESSMENT OF RISK	  II-l

       A.  Toxicity  of TBT to Nontarget Aquatic
            Organisms	  11-2
       B.  Exposure:  TBT  in the Marine and
            Freshwater Environment	  11-13
       C.  Risk Summary	  11-20

III.   BENEFITS	 III-l

      A.  Antifoulant Paint Use Pattern	 III-l
      B.  Tributyltin Antifoulant Paint Registrations ... III-l
      C.  Usage of Tributyltin Antifoulant Paints
            and Alternatives	 III-2
      D.  Benefits of TBT Usage	 III-3

IV.    OTHER REGULATORY CONSIDERATIONS 	  IV-1

      A.  Expansion of Special Review	  IV-1
      B.  Data Call-In Notices	  IV-2

 V.    BIBLIOGRAPHY 	   V-l
                               ii

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  I.     INTRODUCTION

        A.  General Background and Organization


           The Environmental Protection Agency (EPA) is initiating
  a Special Review of all pesticide products containing tributyltin
  (TBT)  active ingredients used as paint additives (antifoulants)
  to inhibit  the growth of certain aquatic organisms.  The TBT
  paints are primarily applied to boat and ship hulls.  These
  TBT compounds include:  bis{tributyltin) oxide, bis(tributyltin)
  adipate, bis(tributyltin) dodecenyl succinate, bis(tributyltin)
  sulfide, tributyltin acetate, tributyltin acrylate, tributyltin
  fluoride/ tributyltin raethacrylate, and tributyltin resinate.
 This Tributyltin Support Document presents the basis for
  initiation of the Special Review on these nine active ingredients.

           The EPA has determined that the pesticidal use of
 these compounds results in TBT exposure to nontarget aquatic
 organisms at concentrations resulting in acute and chronic toxicity
 and,  when applied as antifoulant paints, meet or exceed the risk
 criteria as described in 40 CFR 162.11.  Accordingly, a Special
 Review of products containing these TBT compounds and applied
 as antifoulant paint is appropriate to determine whether additional
 regulatory actions are required.  During the Special Review
 process,  EPA will carefully examine the risks and benefits of
 using TBT products as antifoulants.   If the information reviewed
 indicates that use of these compounds on other sites results in
 exposure  to  nontarget aquatic organisms, the Special Review may
 be extended  to include other pesticidal applications of these
 products  and other TBT active ingredients with registered uses
 which result in  aquatic acute and/or chronic effects or other
 effects of concern.

          This Support Document  contains four parts.  Chapter I is
 this  Introduction.   Chapter II describes the risk to nontarget
 aquatic organisms  resulting from the use of TBT  compounds in
 antifoulant  paints.  Chapter III describes  the usage and  benefits
 of these  TBT compounds.   Chapter IV  outlines other regulatory
 considerations associated with the TBT  compounds.

      B.  Legal Background

          1.  The Statute

              A pesticide product may be sold or distributed in
 the United States only  if it  is  registered  or exempt from regis-
 tration under the Federal Insecticide,  Fungicide,  and  Rodenticide
Act (FIFRA) as amended  (7 U.S.C.  136 et  seq.).   Before  a  product
can be  registered, it must  be shown  that  it  can  be  used without
 "unreasonable adverse effects on the environment"  (FIFRA  section
3(c)(5)), that is, without  causing "any  unreasonable risk to
                               1-1

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 man or  the environment, taking into account the economic,
 social, and environmental costs and benefits of the use of
 the pesticide"  (FIFRA section 2(bb)).  The burden of proving
 that a  pesticide meets this standard for registration is at
 all times, on the proponent of initial or continued registration.
 If at any time  the Agency determines that a pesticide no longer
 meets this standard for registration, then the Administrator
 may cancel the  registration under section 6 of FIFRA.

           2.  The Special Review Process

               The term "Special Review" is the name being used
 for the process previously called the Rebuttable Presumption
 Against Registration (RPAR)  process 40 CFR 162.11.   Modifications
 to the process are described in the final Special Review regu-
 lations (50 FR 49003).  These regulations will become effective
 after 60 days of continuous congressional session after the
 issuance of the regulations, which took place on November 19,
 1985.   These modifications include new risk criteria.  Until
 these  regulations are adopted, the present RPAR procedures
 will remain in effect as set forth in 40 CFR 162.11.

               The Special  Review (RPAR)  process provides a
 mechanism through which the  Agency gathers risk and benefit
 information about pesticides which appear to pose risks of
 adverse effects to human health or the environment which may be
 unreasonable.   Through issuance of notices and support documents,
 the Agency publicly sets forth its position and invites pesticide
 registrants,  USDI,  USDA,  FDA,  user groups, environmental groups,
 and other interested persons to participate in the Agency's
 review  of  suspect pesticides.

               Risk  evidence, submitted to and/or gathered by the
 Agency,  must  be  evaluated  and  considered in light of the benefit
 information.   If the Agency  determines that the risks appear
 to  outweigh  the  benefits,  the  Agency can initiate action
 under FIFRA  to cancel,  suspend,  and/or require modification
 of  the  terms  and conditions  of registration.


      C.   Chem'ical  Background

           1.   Registered Uses  and  Production

               There  are  20 tributyltin compounds registered as
pesticidal active ingredients.  Nine of  the TBT compounds are
registered for use  in antifoulant  paints.   Other registered uses
of the TBT's include but are not limited  to:   use as wood preser-
vatives, textile  biocides, disinfectants,  use  in cooling towers,
paper and pulp mills, and leather  processing  facilities.   Current
annual domestic  production of  TBT  pesticides  for these uses is
estimated at approximately 730,000 to  860,000  pounds of  active
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  ingredient for the 20 TBT active ingredients.  Approximately
  a third of the TBT is used in antifoulant paints, one third is
  used  in wood preservatives, and the remaining third is used
  in other pesticidal formulations.

           2 .  Chemical and Physical Characteristics of TET's in
               Ant if puling Paints

               The tributyltin antifouling paints are chemically
  characterized by a tin (Sn) atom covalently bonded to three butyl
  (C4Hg-) moieties.  A representative TBT active ingredient,
  tri-ri-butyltin fluoride, may be chemically described by the
  following structural formulas for the undissociated (neutral)
 pure form of the active ingredient and for the water dissociated
  (positively charged)  form:
              H9C4 — Sn — C4Hg     HgC4— Sn — €4119
               Elemental or inorganic forms of tin (as in mineral
 deposits or tin cans)  appear to cause negligible toxicological
 effects in man or wildlife.   However, in contrast,  the TBTs
 display an increased fat solubility and consequently, enhanced
 ability to penetrate biological membranes, thereby posing a
 greater toxicity and environmental risk (Thayer 1984).  When
 tributyltin is used as an active ingredient in antifouling paint
 formulations that are  applied for example to boat hulls,  the free
 TBT  ion is leached or  released from the paint, providing fresh
 toxicant at the wetted paint surface to inhibit growth of fouling
 organisms (e.g.,  barnacles, tubeworms ) .

               Table 1  identifies the nine tributyltin active
 ingredients under consideration in the Special Review and their
 Chemical Abstract Service (CAS) number.

               In  general,  the TBT antifouling paints may be
 classified into two categories according to the way  the tributyltin
 moiety  is  incorporated into the paint coating and subsequently
 released.   In  the first group,  the conventional freely associated
 coatings  (e.g., TBT oxide  or fluoride), the biocide  is physically
 incorporated into the  paint matrix (which contains the pigment,
 water-soluble  resins,  and  inert substances).   Upon contact with
 the marine environment,  the surface  oarticles of the paint coating
 are dissolved  with  physical release  of the toxic TBT by diffusion.
 This category  of  TBT antifoulant coatings have traditionally
 posed a problem of  a high  early release rate with subsequently
 shortened  time period  of  protection  from attachment  and growth
of fouling organisms (Fisher e_t al.   1981).   In the  second
category,  the  copoylmer  paints,  the  TBT moiety is chemically
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Table  1.   Tributyltin Active Ingredients as Used in
                 Antifoulant Paints
  Chemical Name

bis(tributyltin) oxide
bis(tributyltin) dodecenyl
  succinate
bis(tributyltin) sulfide
tributyltin acetate
tributyltin acrylate
tributyltin fluoride
tributyltin resinate
tributyltin raethacrylate,
  and copolymer
bis(tributyltin) adipate
Chemical Abstract
  Service Number

  56-35-9
  12379-54-3

  4804-30-4
  56-36-0
  13331-52-7
  1983-10-4
  none assigned
  2155-70-6,
    and 26345-187
  7437-35-6
                        1-4

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 bonded  to  a polymer  backbone  (e.g., TBT methacrylate copolymer).
 This bond  is  designed to be hydrolytically unstable under
 slightly alkaline conditions.  Therefore, the biocide is
 released only by chemical hydrolysis of the tributyltin itself.
 This attachment (pendant polymer) accounts for several advantages:
 1) release is governed by hydrolysis of the TBT group rather
 than dissolution of  the paint particle, 2) the release rate
 can be  better controlled (slowed down) by alteration of the
 polymer's water absorption characteristics, and 3) the formulation
 is safer for  shipyard personnel to handle because the biocide
 is released only when wetted and also the polymeric form poses
 less irritation to skin and nasal passages.  With the exception
 of the  initial high  release rate during the "conditioning"
 period  (approximately the first month after the freshly painted
 hull is placed in the water), these polymeric, film-forming
 resin coatings are characterized by a slow dissolution rate from
 ship hulls and thus  the ability to achieve a constant but prolonged,
 low release rate of antifouling toxicant (ibid.).

               In general, toxicity of organotin compounds to
 aquatic organisms  is thought to increase with the  number of butyl
 substituents  from one to three (Laughlin,  Norlund, and Linden
 1984)  and then to  decrease with the addition of a  fourth butyl
 group.   In order to assess the fate of a particular tributyltin
 derivative in water one  must consider the  dissociated active
 form,  the TBT cation (BujSn"1"), and its major metabolites
 presumably formed  by progressive debutylation to inorganic
 tin (Blunden 1984).

           Bu3Sn"*" —>  Bu^Sn2"1" —> BuSn3"1" —> Sn4+

               In addition,  Matthias and coworkers  (1985)  have
 prepared a  manuscript for publication (currently  in review by
 Analytical  Chemistry) containing their findings of relatively
 high levels  of tetrabutyltin in surface (microlayer)  water samples
 from Baltimore harbor.   This species (Bu4Sn)  could possibly be
 formed  from microbial/photolytic transformation of the tri-
 and dibutyl  forms-which  could  then be reconverted  to  the  dibutyl
 species.  The  tetrabutyltin  species may also be present in
 some paint  formulations  as an  impurity in  the  manufacturing
 process.  Also,  butylmethyltin (BU3MeSn) has  been  isolated
 from marine sediment  samples  (ibid.).

               Analytical  methodology suitable  for  environmental
monitoring must, in view  of  the  foregoing  discussion,  be  able to
detect at low  levels  all  butyltin  and  mixed methylbutyltin species
arising'  from degradation  of  the  TBT  cation in  marine  and  fresh-
water environments.   Furthermore,  because  morphological  and
toxicological  effects have been  found  to occur in  nontarget aquatic
organisms when exposed to TBT  concentrations  in the parts per
trillion (ppt) range, the analytical methodology must  have a
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 corresponding low limit of detection.  A number of speciation
 methods (separation and analysis) have been reported in connection
 with environmental monitoring studies; these have been reviewed
 by Brinckman (1983).  More recently, Brinckman and coworkers have
 prepared a manuscript which presents a simple and rapid method
 for the speciati'on of aquatic samples using simultaneous hydri-
 dization/extraction with separation by gas chromatography and
 detection by flame photometry (Matthias e_t a_l. 1985).   In using
 100 mL salt water samples, the level of detection was  said to be
 7 ppt of tetrabutyltin, 7 ppt of tributyltin, 3 ppt of dibutyltin,
 and 22 ppt of monobutyltin.

               Among the methods cited in this Support  Document in
 connection with environmental monitoring or bioassay studies are
 those that measure the four butyltin species (BunSn) and inorganic
 tin.   They are based upon preliminary extraction and purification
 using an organic solvent (benzene,  tropolone, hexane,  methyl
 isobutylketone)  followed by formation of a volatile derivative
 with  a Grignard reagent (butyl-, pentylor hexylmagnesium bromide)
 or with sodium borohydride (hydride).  Species separation is then
 accomplished by either gas chromatography (GC) or programmed warming
 in a  purge/trap (PT)  system with detection of the volatilized
 organotin  derivative by atomic absorption spectroscopy (AA), flame
 photometric detection (FPD) or mass spectroscopy (MS).  Detection
 limits for tributyltin (cation)  in  marine water columns have been
 reported at 7 ppt (Matthias et al.  1985, using hydridization and
 GC/FPD); 20 ppt (Maguire et aT.  1982, using butyl or pentyl
 derivatives and either GCTfPD or GC/AA); 5 ppt (Valkirs et al.
 1985,  using hydrization and PT/AA);  and, 5 ppt (Huggett 1985,
 using  hexyl derivatives and GC/FPD  with confirmation by MS).  To
 achieve  these detection limits sample volumes have varied from
 500 mL to  2000  mL.

               Since  higher levels of TBT have usually  been observed
 in surface  microlayer,  fish tissues  and  other sorbing  media
 including  sediment,  analytical methods for these specimens have
 been  cited  with  higher detection limits.  These include 60 to 150
ppt for  surface  micro-layers (Maguire 1982,  using 100 mL samples
by GC/FPD);  500-0. ppt  for marine  sediment (Maguire 1982,  using  1
gram dry specimens by  GC/FPD);  and,  80 ppt for fish tissue (Waldock
and Thain  1983,  on tissue  using  flameless AA).
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  II.   ASSESSMENT OF RISK

       The  EPA  has determined that registered products and
  applications for registration of pesticide products containing
  the  following  active  ingredients in antifouling paints:  bis
  (tributyltin)  oxide,  bis(tributyltin) adipate, bis(tributyltin)
  dodecenyl  succinate,  bis(tributyltin) sulfide, tributyltin
  acetate, tributyltin  acrylate, tributyltin fluoride, tributyltin
  methacrylate,  and tributyltin resinate meet or exceed the existing
  criteria (40 CFR 162.11) and new risk criteria (50 PR 49003) for
  acute and  chronic hazard to nontarget aquatic organisms.

       Tributyltin compounds are very effective biocides, toxic to
 marine and freshwater organisms at extremely low concentrations.
 The use of tributyltin compounds in antifoulant paints, has
 resulted in increased concentrations of TBT in harbors, marinas,
 and estuaries.   Environmental hazard to nontarget organisms
  (i.e., nonfouling organisms such as mussels,  clams, and oysters)
 is highly probable,  where TBT concentrations  have been measured
 at or near the  recorded acute and chronic toxicity levels for
 various nontarget aquatic organisms including commercially
 valuable marine organisms.

       EPA has determined that the use of pesticide products
 containing  bis(tributyltin) oxide,  bis(tributyltin) adipate,
 bis(tributyltin)  dodecenyl  succinate,  bis(tributyltin) sulfide,
 tributyltin acetate,  tributyltin acrylate, tributyltin fluoride,
 tributyltin methacrylate, and tributyltin resinate when used in
 antifoulant paints exceeds  the  risk  criterion for acute and
 chronic  toxicity  as  defined in  40 CFR 162.11(a)(3)(i)(B) and 40
 CFR 162.11  (a)(3)(ii)(C)  which  provide that a Special  Review
 shall be  conducted  if  a pesticide:

              "results in a maximum  calculated  concen-
              centration  following  direct application
              to  a 6-inch layer  of water more than 1/2
              the acute LC50 for aquatic organisms
              representative of  the  organisms likely
              to be exposed ...  ."  (40 CFR 162.11(a)(3)(ii)(B))

      Or  if use of the chemical:

              "can" reasonably be  anticipated  to  result
              in significant local regional or  national
              population .reductions  in nontarget
              organisms, or fatality to  members of
              endangered species." (40 CFR 162.11(a)(3)(ii)(C))

      The Agency has issued new Special  Review  regulations  which
include revised risk criteria.  Although  these  criteria  are
not in place at  the present, we have concluded  that these TBT
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 products  also meet  or  exceed the new risk criterion (50 FR
 49003) because concentrations of TBT in the marine and freshwater
 aquatic environments may result

               "in residues of the pesticide product or
               its ingredients, impurities, metabolites, or
               other degradation products in the environ-
               ment of  nontarget organisms at levels which
               equal or exceed concentrations acutely or
               chronically toxic to such organisms . .  .
               at levels which produce adverse reproductive
               effects  in such organisms as determined
               from tests conducted on representative
               species  or from other appropriate data."

       This chapter consists of three sections.  The first
 discusses the toxicity of TBT to nontarget aquatic organisms;
 the second presents the concentrations of TBT which have been
 found in the marine and freshwater environments;  and the third
 section summarizes the potential risk posed by the use of TBT
 active ingredients in antifouling paint formulations.
                                                                   a.
       A.   Toxicity of TBT to Nontarget Aquatic Organisms

           1.  Introduction

               In  this toxicity section, information summarizing
 the adequacy of historical bioassay tests and the acute and
 chronic toxicity  of  TBT to fish,  algae, crustaceans, and molluscs
 will be summarized.

           2.  Measurement of  Toxicity

               Only recently  have  data been available on the
 toxicity  and environmental fate  of  tributyltin compounds,  espe-
 cially with  an  interest toward delineating the relationship  which
 may exist  between the number  of  alkyl groups (i.e.,  tributyl,
 dibutyl, and mono.butyl) and  the  toxicities of these  compounds.
 Many studies were conducted  as static bioassays with nominal non-
 measured  (estimated)  concentrations.   Nominal values do not  take
 into account the  tributyltin  adsorption onto test containers
 resulting  in an overestimate  of the  actual available toxicant
 in  the  solution.   Since the concentration of TBT  toxicant  can  be
 overestimated, the LCso reported may  underestimate the TBT toxicity
 (e.g.,  while  the  actual LC5Q  may be  60  ppt,  the reported  value
may  be  100 ppt since  the  TBT  concentration was not measured).   This
 adsorption problem is critical at very  low levels (i.e.,  < 1.0  ppb)
where a 70 percent decrease  (almost  an  order of magnitude) has
been found between nominal and measured concentrations of
tributlytin  (M &  T Chemical Co., Aug.  1977).
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                Static  renewal  bioassay  testing procedures may
  provide accurate  estimates  of tributyltin concentration if the
  test solution renewal period  is  brief;  this procedure eliminates
  lengthy resident  times and  loss  of  tributyltin by adsorption
  and/or subsequent degradation to intermediate forms.  Flowthrough
  bioassay testing  with simultaneous  chemical analyses for chemical
  speciation  of the toxicant  in solution  is, however, a more
  desirable testing approach.

                The detection limits  of TBT concentration measurement
  are  of major  concern.   TBT has been detected in water column
  samples using the borohydride method (Matthias et al. 1985) as
  low  as 7  ppt.  This method is most  effective with large water
  sample sizes.  The Virginia Institute of Marine Science (VIMS)
  has  been  able to  detect levels of tributyltin as low as 2.0 ppt
  (Perkins  1985).   The  determination of environmental concentrations
  in water  column samples is critical to the development of a risk
  assessment.   The  inability to measure low ppt levels of TBT can
  result  in an  erroneous assessment of hazard at the sublethal
  chronic  level.

               The  following discussion characterizes the concerns
  EPA has regarding  tributyltin toxicity to nontarget aquatic
 organisms subsequent to their exposure to TBT concentrations now
  found  in harbors, marinas, lakes, and estuaries.   Table 2
 summarizes the TBT toxicity data used to initiate the Special
 Review.  It should be noted that different investigators used
 different chemical forms (salts) of TBT in conducting the aquatic
 toxicity studies.   These included the chloride,  the bis(tributyltin)
 oxide and the methacrylate.  Although LCsg values were reported
 using these  different forms of TBT, their effect  on the final
 measured or  calculated result  is relatively negligible.   For
.example, the molecular weight  ratio between the  bis(tributyltin)
 oxide and tributyltin chloride is only two-fold,  whereas,  the
 acute toxicities  vary  across species of fish  by  approximately
 24-fold.

           3.   Toxicity to  Fish

               Acute toxicity testing usually  measures the  lethal
 (LC5o)  or effective (EC$Q)  concentrations where  50  percent  of the
 test  organisms are killed  or significantly affected.   Acute
 toxicity of  bis(tributyltin) oxide (TBTO)  to  certain  freshwater
 fish  appears  to range  from  0.96 ppb to  24.0 ppb.   Reported  96-hour
LC5Q  values  (nominal concentrations)  for rainbow  trout  (Salmo
gairdneri), bluegill  (Lepomis  macrochirus), channel cat  fish
 (Ictalurus punctatus),  and mummichoq (Fundulus heteroclitus)  were
6.9,  7.6,  12.0, and 24 ppb,  respectively (M & T Chemical Co.
Sept. 1976; Oct. 1976;  June  1978).   These  studies were  static
nonrenewal, using  nominal concentrations,  and may not be sensitive
indicators of  actual toxicity  (i.e.,  actual LC5Q  levels  could be
70 percent lower due to  adsorption  to test container  surfaces and
volatilization).
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Table 2.  Test Conditions, Procedures, and Results of Tributyltin Tpxicity Tests
Test Organism
FISH
Lepomis
macrochirus
(Bluegill)
Salmo gairdneri
(Rainbow trout)

Ictalurus
punctatus
(Channel catfish)
Fundulus
heteroclitus
(Mumnichog)
Salmo gairdneri
(Rainbow trout)














Organotin
Compound

TBTO


TBTO


TBTO


TBTO


TBTO















Effect

96-hr LCso = 7.6 ppb
(5.6 to 10 ppb)

96-hr LC50 = 6.9 ppb
(6.27 to 7.8 ppb)

96-hr LC50 = 12.0 ppb
(7.3 to 20.0 ppb)

96-hr LCso =24.0 ppb


24 hr EC5o = 31.0 ppb
(loss of positive
rheotaxis). Level from
5850 ppb to 11.7 ppb.
TBTO resulted in damage
to gill epithelium. At
11.7 ppb there was a
flattening of bile duct
columnar epithelial
cells and separation
from connective tissue
after 5-day exposure.
Destruction of corneal
epithelium occurred
after 7-day exposure
to 11.7 ppb.
Analytical
Methods

Nominal
concentra t ions

Nominal
concent rat ions

Nominal
concentrations

Nominal
concentrations

Not reported















Type of
Exposure

Static


Static


Static


Static


Continuous
flow stain-
less steel
tanks












Concentration
Levels

5.6, 7.5, 10.0 and
14.0 ppb

4.1, 5.3, 6.8, 8.8,
and 11.0 ppb

7.5, 14.0, 18.0,
24.0 and 28.0
ppb
32.0, 42.0, and
56.0 ppb

5850 to 11.7 ppb
acetone














Reference

M & T
Chemical Co.
(Oct. 1976)
M & T
Chemical Co.
(June 1978)
M & T
Chemical Co.
(Sept. 1976)
M & T
Chemical Co.
(Sept. 1976)

Chi iamov itch
& Kuhn
(1976)













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Table 2.  Test Conditions, Procedures, and Results of Tributyltin Toxicity Tests (Continued)
Test Organism
Cyprinodon
variegatus
(Sheepshead
minnow)
33 to 49 ram
Cyprinodon
variegatus
( Sheepshead
minnow)
17 to 25 mm



.

Salmo gairdneri
(Rainbow trout
yolk sac fry)










Organotin
Compound
TBTO




14C-TBTO








TBTCL












Effect
21-day LC50 = 0.96 ppb
Total mortality at
3.2 ppb at 14 days.


The maximum observed
bioconcentration fac-
tors were X2120 and
X4580 for the head and
viscera, respectively.
After 58 days the de-
puration of 14C-TBTO
from all tissues was
rapid. (52% after 7
days).
10 to 12 day LC^oO =
5.0 ppb. Decrease in
growth rate at 0.2 and
1.0 ppb for 110 days.
Toxicity not observed
at 0.2 and 1.0 ppb;
hemoglobin concentra-
tion and erythrocyte
count were reduced in
blood; hyperplasia and
diminished glycogen
content observed in
liver.
Analytical
Methods
AAS measured
concentrations



LSC measured
concentrat ions







Nominal
concentrations











Type of
Exposure
21-day
flow-
through
acute
testing
Exposed to
TBTO for
58 days.







110-day
continuous-
flow expo-
sure









Concentration
Levels
0.33, 0.63, 0.70,
1.5, 3.2
ace tone-me thanol


0.96 to 2.07 ppb
acetone







0, 0.2, 1.0, and
5.0 ppb











Reference
Ward
et al.
(1981)


Ward
et al.
(1981)







Seinen
et al.
(1981)











-------
                 Table; 2.  Test Conditions, Procedures, and Results of Tributyltin Toxicity Tests (Continued)
   Test Organism
Organotin
Compound
        Effect
                         Analytical
                           Methods
            Type of
            Exposure
               Concentration
                   Levels
                     Reference
   ALGAE
   Ankistrodesmus
     falcatus
   (Freshwater algae)
   Skeletonema
     costatum
   (Marine diatoms)
y  Thalassiosira
<*    pseudonana
   (Marine diatoms)
  TBTO
  TBTO
A maximum algal bio-
concentration factor
of 3 x 104 was esti-
mated for TBT after
7 days.

72 hr EC50 = 0.33 ppb
                          GC and FPD
             Static
             20 ppb
             methanol
                             ICAP
   TBTO
72 hr
           = 1.03 ppb
ICAP
Static
Nominal
(only
stock
measured)

Static
Nominal
(only
stock
measured)
                        0.5, 1.0, 5.0, 7.5,
                           10.0, 15.0, and
                           25.0 ppb
0.5, 1.0, 5.0,  7.5,
  10.0, 15.0, and
  25.0 ppb
                    Maguire
                    et al.
                      (1984)
                    Walsh
                    et al.
                      (1985)
Walsh
et al.
 (1985)

-------
Table 2.. Test Conditions',  Procedures, "'and Results of Tributyltin Toxicity Tests (Continued)
Test Organism
CRUSTACEANS
Crangon crangon
(Shrimp)

Acartia tonsa
(Copepod)



Homarus americanus
M (Lobster larvae)
i
>j
Rhithroponopeus
harrisii
(Mud crab)








Daphnia magna
(Water flea)
Organotin
Compound

TBTO


TBTO




TBTO


TBTO










TBTO
Effect

Larvae 96 hr IC$Q =
1.5 ppb. Adult 96 hour
LC5Q =41 ppb.
72 hr EC5o =2.1 ppb
96 hr ECso =1.0 ppb
144 hr ECso =0.4 ppb


90% decrease in growth
at 1.0 ppb.

Bioaccumulation of
4400X in the hepato-
pancreas. No steady
state achieved. Accu-
mulation of TBTO fron
food greater than from
only the water




48 hr LC5o = 1.67 ppb
(1.01 to 2.5 ppb)
Analytical
Methods

Not reported


Measured con-
centrations
AAS


Nominal
concentra t ions

Measured con-
centration of
1 A m
C14 radio-
assay.







Nominal
concentration
Type of
Exposure

Static re-
newal every
24 hours
Static re-
newal every
24 hours
Pyrex test
tubes
Static re-
newal every
48 hours
Static re-
newal every
24 hours.
Artemia
with con-
centrations
1.23 ug
TBTO/g wet
weight were
fed to
crabs.
Static
Concentration
Levels

Not reported


0.3, 0.5, 1.0, 1.7,
and 3.0 ppb
acetone


1.0, 5.0, 10.0,
15.0, and 20.0
ppb
0.28 ppb and
6.1 ppb









1.7, 3.0, 5.3, 7.1,
and 13.3 ppb
ethanol
Reference

Thain
(1983)

U'ren
(1983)



Laughlin &
French
(1980)
Evans &
Laughlin
(1984)








M & T
Chemical Co
(Jan. 1976)

-------
Table 2.  Test Conditions, Procedures, and Results of Tributyltin Toxicity Tests (Continued)
Test Organism
MOLLUSCS
Crassostrea
virginica
(Eastern oyster
larvae)
Crassostrea gigas
(Pacific oyster
larvae)
Mytilus edulis
(Mussel larvae)
M
H
oo Mytilus edulis
(Mussel larvae)
Crassostrea gigas
(Pacific oyster
spat)
Mytilus edulis
(Mussel spat)
Organotin
Compound
TBTO
TBTO
TBTO
TBTO
Exposure for
45 days to
tributyltin
methacrylate
leachates
Exposure for
45 days to
tributyltin.
roe thacry late
leachates
Effect
48 hr EC5Q =0.9 ppb
(0.4 to 1.9 ppb)
48 hr LC5o =1.6 ppb
48 hr LCso =2.3 ppb
15 day LCso =0.1 ppb
long-term effect
Significant reduction
in growth at 0.24 ppb
Significant reduction
in growth at 0.24 ppb
Analytical
Methods
Nominal
Concentration
Not reported
Not reported
Measured con-
centrations
FAAS
Measured daily
for 3 weeks
and every
other day for
the remainder
of the experi-
ment (M&T
standard
method)
Measured daily
for 3 weeks
and every
other day for
the remainder
Type of
Exposure
Static
Static re-
newal every
24 hours
Static re-
newal every
24 hours
Static re-
newal every
72 hours
Plow-
through
Flow-
through
Concentration
Levels
0.1, 0.3, 0.6, 1.0,
3.2, and 5.6 ppb
Not reported
Not reported
10.0, 1.0, and 0.1
ppb
0.24 + 0.23 ppb
for low-level and
2.62 + 1.09 ppb
for hTgh level
0.24 + 0.23 ppb
for low-level and
2.62 + 1.09 ppb
for high level
Reference
M&T
Chemical Co
(June 1977)
Thain
(1983)
Thain
(1983)
Beaumont
& Budd
(1984)
Thain &
Waldock
(1985)
Thain fi
Waldock
(1985)

-------
Table  2o   Test Cond it ions,.. Procedures, and Results of Tributyltin Toxicity ies>i.&  uLinueu)
Test Organism
Mytilus edulis
(Mussel spat)
(Continued)

Venerupis
decussata
(Clam spat)






M
M
*> Ostrea edulis
(European oyster
;.spat)



<•


Venerupis
semidecussata
(Clam spat)





Organotin
Compound




Exposure foe
45 days to
tributyltin
methacrylate
leachates






Exposure for
45 days to
tributyltin
methacrylate
leachates




Exposure for
45 days to
tributyltin
methacrylate
leachates




Effect




Significant reduction
in growth at 0.24 ppb









Growth inhibited at
2.6 ppb






Growth inhibited at
2.6 ppb





Analytical
Methods
of the experi-
ment (M&T
standard
method)
Measured daily
for 3 weeks
and every
other day for
the remainder
of the experi-
ment (M&T
standard
method).


Measured daily
for 3 weeks
and every
other day for
the remainder
of the experi-
ment (M&T
standard
method).
Measured daily
for 3 weeks
and every
other day for
the remainder
of the experi-
ment (M&T
standard
method).
Type of
Exposure




Flow-
through









Flow-
through






Flow-
through





Concentra t ion
Levels




0.24 + 0.23 ppb
for low-level and
2.62 + 1.09 ppb
for hTgh level







0.24 + 0.23 ppb
for low-level and
2.62 + 1.09 ppb
for high level





0.24 + 0.23 ppb
for low-level and
2.62 + 1.09 ppb
for high level





Reference




Thain &
Waldock
(1985)








Thain &
Waldock
(1985)






Thain &
Waldock
(1985)






-------
              Table 2.  Test Conditions, Procedures, and Results of Tributyltin Toxicity Tests -(Continued)
Test Organism
Organotin
Compound
        Effect
 Analytical
   Methods
 Type of
 Exposure
  Concentration
      Levels
 Refere
Ostrea edulis
(European oyster-
  spa t)
  TBTO
Crassostrea qigas
  TBTO
 (Pacific  oyster
Ostrea edulis
.(European oyster)
  TBTO
 Crassostrea gigas
 (Pacific oyster
   spat)
   TBTO
 Nassarius
  obsoletus
 (Mud snail)
 Alumacide
 TBT paint
Growth effected at
0.02 ppb (marginal).
At 0.06 ppb growth
rate was severely
curtailed after 10
days.

Bioaccumulation of
2000 to 6000 fold after
22 days (poor diet
was noted).

Bioaccurnulation of
1000 to 1500 fold
after 22 days (poor
diet was noted).
Spat grew poorly at
TBT  concentrations of
0.15 ppb; developed
pronounced  thickening
of upper shell  valve.
Bioconcentration
 (Flesh)  after 56 days
exposure to 0.15 ppb
was about XI1400 fold.

Anatomical  abnormality
consisting  of super-
 imposition  of male
 characteristics on
 female snails.
Nominal con-
centration
renewed every
day.  (M&T
standard
method).

FAAS measured
for the first
5 days of
exposure and

FAAS measured
for the first
5 days of
exposure and
on alternate
days until the
completion.

FAAS measured
(M&T Standard
method)
 Static
 renewal
 24 hours
Flow-
 through
Flow-
 through
Static re-
newal 24
hours
0.02 to 2.0 ppb
0.15 and 1.25 ppb
0.15 and 1.25 ppb
0.08 to 1.2 ppb
                                                            Not reported
               Not reported
             Not reported
Thain &
Waldock
(1985)
Waldock
Thain,
Miller
 (1983)

Waldock
Thain,
Miller
 (1983)
Waldock
Thain
 (1983)
                     Smith
                      (1981)
 QC   ~ Gas Chromatography
 FPD  = Flame Photometric Detection
 ICAP = Inductively Coupled Argon Plasma
                                    FAAS = Flame less AU*nic .Absorption Spectrophotometry
                                    ISC  = Liquid Scintillation Counting

-------
               Ward  (Ward et al. 1981) found that sheepshead minnow
  (Cyprinodon variegatus) exposed to TBTO for 21 days (measured
  flowthrough system) had an LC$Q of 0.96 ppb and total mortality
  in 14 days at a concentration of 3.2 ppb.  It was further noted
  that after exposure to TBTO (0.96 to 2.07 ppb) for 58 days, the
  maximum observed bioconcentration factors were 2120X and 4580X
  for head and viscera, respectively.  He also noted that after an
  additional 58 days  (i.e./ 116 days total) the depuration, or
  elimination, of TBTO from all tissue was rapid (52% after 7 days
  post-exposure).

               Chronic effects on fish were found by Seinen et al.
  (1981) after subjecting rainbow trout yolk sac fry to 110 day
  exposure to concentrations of tributyltin chloride (continuous-flow
  exposure, nominal concentrations).   A 10 to 12 day LC^oo °f 5
 ppb was estimated using concentration levels 0.2,  1,  and 5 ppb.
 Although, young trout exposed to tributyltin chloride concen-
 trations of 0.2 and 0.1 ppb did not exhibit mortality,  there was
 a significant reduction in growth (40% decrease in body weight),
 decrease in hemoglobin content, hyperplasia of liver  cells, and
 diminished glycogen content in liver.  Chilamovitch and Kuhn
  (1976)  noted histological  and hematological effects on  rainbow
 trout after continuous exposure to  TBTO.  Damage to gill epithelium
 was  recorded at 1.7 ppb.   At 11.7 ppb with 5  days  of  exposure,
 there was a flattening of  bile duct columnar  epithelial cells and
 separation of  these cells  from connective tissue.   Destruction of
 corneal  epithelium was observed following 7 days of exposure to
 11.7  ppb.

           4.   Toxicity to  Algae

               The  toxicity of  tributyltin compounds to  algae has
 not been  intensively studied.    Maguire  et al.  (1984) noted that
 the freshwater algae (Ankistrodesmus  falcatus)  possesses a mechanism
 for sequential debutylation  of  TBTO.  However,  Walsh  et al.  (1985)
 found that TBTO  inhibited  population  growth and cell  survival of
 marine unicellular  algae Skeletonema  costatum  and  Thalassiosira
 pseudonana at  low concentrations  (72  hour  EC5Q  of  0.33  ppb and
 EC5Q of 1.03 ppb, respectively).

          5.  Toxicity  to Crustaceans

              The toxicity of tributyltin  to crustaceans was
evaluated by several researchers.  Laughlin (1980)  using nominal
concentrations in a  static daily renewal,  found  that  lobster larvae
 (Homarus americanus) exhibited a 90 percent decrease  in growth
at 1 ppb.  U'ren (1983) calculated a  144 hour EC^Q  for  marine
copepods (Acartia tonsa) at 0.4 ppb (measured concentrations
static daily renewal).  Thain (1983)  reported 96 hours  LC5o  values
for larvae and adult shrimp (Crangon  crangon) at 1.5 and 41  ppb,
respectively.
                              11-11

-------
                Tributyltin  bioaccuraulation  in crustaceans was
  demonstrated  by  Evans  and  Laughlin  (1934)  with thair work on mud
  crabs  (Rhithroponopeus harrisii) exposed to radiocarbon labeled
  TBTO.   The  test  measured the  short-term effects of exposure to
  labeled test  concentrations in water  (0.28 ppb) and food (Artemia,
  1.23 ppb)o  Four-day bioaccumulation  factors ranged up to 4400 in
  the hepatopancreas with no indication that a steady state equili-
  brium  had been approached.  Accumulation of tributyltin from food
  was greater than that  from the water.

           6°  Toxicity to  Molluscs

               Acute toxicity  of TBTO  to certain molluscs appears
  to range from 0.1 to 2.3 ppb.  Reported 48-hour LC$Q values for
  Eastern oyster larvae  (C_.  virginica), Pacific oyster larvae
  (C_. gigas)  and mussel  larvae  (M. edulis) were 0.9, 1.6 and 2.3
 ppb, respectively (M & T Chemical Co. June 1977;  and Thain
  1983).  A 15-day LC^Q  of 0.1  ppb was  also reported for mussel
 larvae  (M. edulis) (Beaumont  and Budd 1984).

               Anatomical abnormalities attributed to tributytina
 were found  in certain  intertidal mud  snails (Massarius obsoletus)
 living near yacht basins (Long Island Sound and Southport,  CT).
 Smith (1981),  by investigating this phenomenon it was noted that
 these dioecious snails had developed male characteristics on
 apparently normal female reproductive anatomy.   The correlation
 between tributyltin concentration and these abnormalities was
 later confirmed in laboratory studies.

               European researchers  have found a significant corre-
 lation between the TBTO concentration levels  in certain estuarine
 areas  and shellfish deformity. Waldock and  Thain  (1983) in England
 found  that Pacific oyster (Crassostrea gigas)  spat (set oyster
 larvae) grew poorly in TBTO concentrations  of 0.15 ppb, and
 developed pronounced  thickening of  the upper  shell valve.   Bio-
 concentration  factors,  after  56 days exposure to  0.15 ppb TBTO,
 were 11,400X.   Alzieu  et al_«_  (1980)  observed  similar malformations
 in  C.  gigas  and attributed  it  to the presence of  organotin (TBTO
 and TBTF)  in the  French shellfish regions.  Beaumont and Budd
 (1984)  noted that common mussel (Mytilus edulis)  larvae did not
 survive past 5  days at  a TBTO  exposure concentration of 10  ppb,
 or  longer  than  10 days  at  a TBTO level of  1 ppb.   They concluded
 that the high  TBT levels found at several estuarine sites  are
 associated with adult  shellfish population  reductions  in these
 areas  (a 15  day LC^Q of 0.1 ppb was  estimated).   Thain and  Waldock
 (1985)  tested several shellfish larvae with tributyltin methacrylate
 leachates in measured flow-through systems.  They  found that  Pacific
oyster  (Crassostrea gigas), mussel  (Mytilus edulis),  and clam
 (Venerupis decussata) showed a significant  reduction  in growth at
0.24 ppbo  A static renewal (nominal concentrations)  test with
spat of  the  European flat oyster  (Ostrea edulis)  demonstrated
that growth  rate  was severely  curtailed  following  exposure  to
0.06 ppb TBTO.
                               11-12

-------
               According to Waldock and Thain (1983), oysters (C.
 gigas and O. edulis) rapidly bioaccumulate TBT and reach an
 equilibrium uptake after exposure, and subsequently are slow to
 depurate this .uptake.  Using measured concentrations in a 21-day
 continuous-flow system,  Waldock and Thain exposed £. gigas and
 O. edulis (10 g wet weight) to TBTO concentrations of 1.25 and
 0.15 ppb.  C. gigas accumulated a high body level of tributyltin
 that ranged from 2000- to 6000-fold.  O. edulis maintained under
 the same conditions only concentrated tributyltin from 1000- to
 1500-fold.  This twofold to fourfold difference in tissue concen-
 tration of tributyltin between species correlates with the threefold
 to .ninefold difference found in the field.

               The probability of food chain accumulation was
 addressed by Laughlin, French, and Guard (1984), studying
 tributyltin uptake by marine mussels (Mytilus sp.).   TBT
 bioaccumulate.d from 1000 to 6000 times and was  attributed more
 to food intake than to direct absorption from water or sediment.

               Given its high lipophilicity TBT  is likely to be
 bioaccumulated by several organisms.   Laboratory studies may
 underestimate the extent of bioaccumulation as  they are generally
 of insufficient duration to allow all compartments within the
 test  vessel  (food material, feces,  water,  test  organisms, vessel
 walls,  etc.)  to reach equilibrium.   Studies are needed to evaluate
 the subsequent toxicity problems associated with high bioaccumu-
 lation  of  TBT.

      B.   Exposure:   TBT in the Marine and Freshwater Environment

           In  addition to review of  available bioassay and aquatic
 toxicity data,  the  Agency has also  evaluated data available on
 TBT concentrations  in water samples analyzed from both marine and
 freshwater environments.   Analytical  methods sensitive to the ppt
 level have only  recently been developed  and monitoring data using
 this  new methodology  are currently  very  limited.   Some of this
 information is shown  in Table 3.  Data from Canadian harbors in
 Lake Superior  and Lake  Ontario are  included to  provide some
 indication of  the likely  contamination levels in  Great Lakes
 harbors and other freshwater  bodies with similar  water craft use
 patterns.

          The data  from San Diego Bay, at  present,  is one of the
most systematic analyses of tributyltin  levels  in a  U.S.  bay or
estuary (Valkirs et al. ,  1985).  On the  basis of  informal surveys
of local retail outlets, Valkirs and coworkers  estimated  that the
use of tributyltin on recreational  craft in this  bay greatly
increased during the course of  the  study.   This appears  to be
reflected by a sharp elevation  in the  concentration  of  TBT in the
waters near Shelter Island Yacht harbor where a study maximum of
930 ppt tributyltin was measured.   This  investigation in  the
Shelter island waters is particularly  significant  because


                              11-13

-------
 antifouling  paints  are  likely  to have been the only source of
 tributyltin  in  that area  (i.e. there are no drydock T3T discharges
 or other TBT point  discharges  identified in the area).  In relation
 to the entire San Diego Bay study, it should be noted that,
 although tributyltin  is thought to readily bind to sediment,
 (U.S. Naval  Sea Systems Command 1984) divers taking discrete
 water samples 10 cm above the  bottom sediments found approximately
 the same concentrations as were detected in surface water samples.
 Figure 1 identifies the location of the sampling stations.

           The monitoring data  discussed above may not be totally
 comparable to values  (LCso's etc.) reported in aquatic toxicity
 studieso  Most aquatic  toxicity studies are conducted with
 filtered water into which the  toxin has been initially dissolved.
 The monitoring data reviewed above represent unfiltered water
 analyses which would, perhaps, include sediment bound toxin and
 even, perhaps, tributyltin which had been assimilated into plank-
 tonic organisms.  Further investigation as to whether or not a
 large proportion of aqueous TBT would be in a bound form is
 needed,,   If binding is  found to be significant then the toxico-
 logical  significance will have to be determined.

           There is also reason to believe that the monitoring data
 might underestimate the hazard.  The Great Lakes  and Chesapeake
 Bay researchers with whom the Agency has been in  direct contact
 state that  none of their samples have been taken  in the late
 spring when freshly painted boats would be launched.   Monitoring
 by Waldock  and Miller (1983)  in an English estuary found highest
 annual concentrations in May (corresponding to the postwinter
 launching of  pleasure craft).

          In  addition to the samples noted in Table 3 are samples
 collected by  Virginia Institute of Marine Science (VIMS)  upstream
.and downstream (as influenced by tidal  action)  from large
 commercial  ships moored  near  the Elizabeth River  in Hampton Roads.
 The concentration of tributyltin above  the ships  was  approximately
 5  ppt.  The concentration  in  the surface water  downstream from
 ships  was 13  ppt.   Figure  2  identifies  the sampling stations used
 for the VIMS  one day study.

          Attempts  were  made  during  the  Annapolis and Great Lake
 studies to measure, the concentration  of  tributyltin in the
 surface microlayer—where  high  concentrations  of  lipophilic
 pesticides have  been found, as  noted  from other studies in the
 literature.   Contamination of  the  water  surface is presumed to be
 injurious to  floating  fish eggs such  as  croaker,  herring,  and
 shad species.  Unfortunately no completely  satisfactory method of
 sampling such a  thin  layer of the water  column  is  available.
                               11-14

-------
Table 3.  Tributyltin Concentrations (ng/L = ppt) for Water Samples )V Collected
               at Indicated Stations with Analytical Method Noted
Location
Annapolis, MD
Across harbor
from city docks
Marina in Back
Creek
Lake Superior
(Marathon)
Lake Ontario
(Toronto Harbor)
Lake Ontario
(Hamilton Harbor)
Lake Ontario
(Whitby Harbor)
San Diego Bay2/
Station 3-2
3 Jan 83


Station 3-3
3 Jan 83
n
14 Feb 84
Station 3-4
3 Jan 83
ii
Station 4
3 Jan 83
n
14 Feb 84
Station 5
3 Jan 83
n
14 Feb 84
!an Diego Bay
Station 1
3 Jan 83

n
Concentration
(pp^TBT*)
34


71

20

840

160

50

. •

50

130

100
550
180

'10
110

ND
30
30

ND
ND
10


ND

10
Sample Type
Depth (Meters)
1


1

5

5

5

5



0.3-0.6

BottonV

0.3-0.6
Bottom
0.3-0.6

0.3-0.6
Bottom

0.3-0.6
Bottom
0.3-0.6

0.3-0.6
Bottom
0.3-0.6


0.3-0.6

Bottom
Analytical
Method
Hydride deriviti-
zation, GC/FPD

Hydride deriviti-
zation, GC/FPD
Pentyl deriviti-
zation GC/FPD
Pentyl deriviti-
zation GC/FPD
Pentyl deriviti-
zation GC/FPD
n



Hydride deriviti-
zation, AA4/
n

n

u

n
n

n
n
ii

«
n
n


Hydride derviti-
zation, AA
n
Ref.
1


1

2

2

2

2



3

.3

3

3

3
3

3
3
3

3
3
3


3

3
                                      11-15

-------
Table 3.  Tributyltin Concentrations (ng/L = ppt) for Water Samples^/ Collected
             at Indicated Stations with Analytical Method Noted (Continued)
Location
San Diego (Cont.)
Station 2-1
3 Jan 83

n
20 Jun 84
7 Mar 85
25 Sep 85
Station 2-2
3 Jan 83
H
14 Feb 84
2 Jul 85
25 Sep 85
Station 2-3
3 Jan 83
n
14 Feb 84
Station 3-1
3 Jan 83
n
14 Feb 84
18 Dec 84
Elizabeth River, Near
Norfolk, Va5/
16 Sep 1985
Station 1

Station 2
Station 3
Station 4
Station 5
Station 6
Station 7
Station 8
Concentration
(ppt TOT*)


60

100
270
290
780

50
60
350
490
930

20
40
200

180
140
200
.210



6

6
7
12
21
63
49
39
Sample Type
Depth (Meters)


0.3-0.6

Bottom
0.3-0.6
0.3-0.6
0.3-0.6

0.3-0.6
Bottom
0.3-0.6
0.3-0.6
0.3-0*6

0.3-0.6
Bottom
0.3-0.6

0.3-0.6
Bottom
0.3-0.6
0.3-0.6



1

1
n
n
n
n
n
n
Analytical
Method


Hydride deriviti-
zation, AA
n
n
n
•

n
n
it
n
n

n
n
n

H
n
n
n



Hexyl deriviti-
zation
n
N
n
n
n
n
n
Ref.


3

3
3
3
3

3
3
3
3
-i
..i

3
3
3

3
3
3
3



4

4
4
4
4
4
4
4
                                   11-16

-------
I/  All water samples were whole.  No filtered samples were made.
2/  See Figure 1 for location of sampling stations.
3/  Bottom samples were taken 10 cm from the bottom by scuba divers.
4/  AA Hydrogen flame atonic absorption spectrophqmetry.
5/  See Figure 2 for location of sampling stations.

    References

    1.  Matthias et al.  1985.
    2.  Macguire et al.  1982.
    3.  Valkirs  et al. 1985.
    4.  Perkins  1985.
                                     . 11-17

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        Figure i.   Navy sampling  Stations in  San Diego Bay!/
                      3. COMMERCIAL BASIN
         2. SHELTER ISLAND
           YACHT BASIN
                                            4. COMMERCIAL
                                              ORYOOCX
                                                 5. NAVAL STATION
             1. COAST
               GUARD
               STATION
                              COMMERCIAL
                                BASIN
       YACHT REPAIR
          FACILITIES
Valkirs et aK  1985.
                             .11-18

-------
     rxyure ^.  Virginia Institute of Marine Science Sampling
               Stations in the  Elizabeth River Estuary!/
]_/ R.J. Huggett, Virginia Institute of Marine Science

                               11-19

-------
 Both the Annapolis and Great Lakes studies indicated,  however/
 that the concentration is much higher in the microlayer  and
 the Agency is concerned about risks that this contamination
 may pose.

       C.  :Risk Summary

           After extensive evaluation of published reviews,
 consultation with various Federal and State laboratories,  and
 review of EPA data files, EPA has concluded that tributyltin
 (TBT)  in antifouling paints may be a potential hazard  to nontarget
 aquatic organisms in areas of high boat traffic, marinas,  and
 estuaries.  The toxicity and threat of TBT exposure to aquatic
 organisms satisfies the existing risk criteria (40 CFR 162.11)
 and the new risk criteria (50 FR 49003)  for acute and  chronic
 hazards to nontarget aquatic organisms as defined earlier  in this
 chapter.   A summary of aquatic toxicity values is presented as
 follows:
           1)   Molluscs:   Acute LCso = 0.1  to 2.3 ppb
                       -v  Chronic effects:   0.06  to 0.24  ppb
                          Bioaccumulation:   2000- to 11,000-fold;
                            very slow depuration.

           2)   Fish:   Acute LC50 = 0.96 to  24.0 ppb
                      Chronic effects:  0.2 to 10.0 ppb
                      Bioaccumulation:  2120- to   4580-fold;  rapid
                       depuration.

           3)   Crustaceans:   Acute LC$Q = 0.3 to  41.0 ppb
                             Chronic effects:  1.0 ppb
                             Bioaccumulation:  4400-fold;  greater
                               accumulation from  food than from water.

           4).   Algae:  .Acute LC$Q =  0.33 to 1.03  ppb
                       Bioaccumulation:  8000- to 30,000-fold;
                        toxic  in some species, depuration in
                        others.

           Environmental concentrations of  tributyltin are listed in
Table 3.   Several locations appear  to have tributyltin concen-
trations equal 'to, or  greater  than,  toxicity values for  nontarget
aquatic organisms.  The following conclusions may be made regarding
toxicity and exposure  at  some  of the  locations:

           1)  Annapolis:   tributyltin concentrations of  .034
                           to .071 ppb may  cause  acute mollusc,
                           and  chronic mollusc effects.
                              11-20

-------
            2)  San Diego Bay:  tributyltin concentrations of
                               .01 to  .93 ppb may cause acute
                               and chronic effects in several
                               aquatic taxa (i.e., fish, mollusc,
                               crustaceans, algae).

            3)  Lake Superior:  tributyltin concentrations of .02
                               ppb may cause chronic mollusc
                               effects.

            4)  Lake Ontario:  tributyltin concentrations of .05
                              to .84 ppb may cause acute and
                              chronic effects in several aquatic
                              taxa (i.e., fish,  mollusc, crusta-
                              ceans, algae).

           5)  Norfolk, VA:  tributyltin concentrations of .006
                             to .06 ppb may cause chronic mollusc
                             effects.

           EPA is concerned about the acute and  chronic toxicity
 potential of tributyltin compounds to nontarget aquatic
 organisms.  Water samples have been found to contain TBT
 levels that may  have direct affects on aquatic  organism popu-
 lations (molluscs).   The TBT compounds may bioaccumulate in
 aquatic biota and may pose a hazard to the food chain as they
 are passed from  lower to higher  trophic levels.  Adsorption
 of  tributyltin compounds to sediment may have long-term toxicity
 effects on benthic browsing organisms (crustaceans,  snails,
 etc.).   Contamination of estuarine areas at  sublethal concen-
 trations can influence fecundity of several  aquatic  taxa from
 fish to zooplankton,  thus  influencing population dynamics.
 The present use  of tributyltin in antifouling paints presents
 a potential hazard to nontarget  aquatic  organisms.

           The tributyltin  compounds are  acutely and  chronically
 toxic  to molluscs  at  low levels  (0.06  to 2.3  ppb)  depending
 upon molluscs species.   A  correlation between environmental
 concentrations of  tributyltin  and  declining populations of
 nontarget organisms  has  been demonstrated in  Europe  through
 the  works  of  Waldock  and Thain,  Beaumont,  and Alzieu.   Laboratory
 toxicity testing,,  environmental  monitoring, and bioaccumulation
 studies  have  corroborated  the  conclusion that tributyltin
 antifouling paints may be  responsible  for the decline of  several
 mollusc  populations.

           Based on these findings, Great Britain and France
have regulated or curtailed  the  use  of tributyltins  in  antifouling
paints.   Effective January  1,  1986 Great Britain is  banning:
a)  copolymer  formulations containing more  that  7.5%  organotin
 (measured  as  tin in the dry  paint  film)  and b)  those paints
                            11-21

-------
 based on copper or other antifouling systems  containing  more
 than 2.5% organotin (measured in the same  way).   In  effect  (b)
 will ban the supply of existing  "free association" paints while
 allowing the minimum use of  tributyltin  compounds as a performance
 booster in other antifouling systems Great Britain will  review
 these levels in time for the 1987  painting season with a view
 to  reducing them in line with advances in  paint  technology.  In
 addition, the British government proposes  to  establish an
 ambient water quality target for organotin concentrations.

           France has recently extended its ban another 2 years
 on  use  of organotin paints for all boats less than 25 meters in
 length.   The French experience shows that  a ban  on the use of
 TBT on  pleasure craft can be highly effective.   In 1982  the
 industry producing  organotin compounds measured  concentrations
 in  Arcachon Bay and found levels 3 times higher  than those
 known to cause malformation  in Pacific Oysters.  Since the
 implementation of  the first  ban  (1983  to 1985),  the  recovery
 of  oysters  has been carefully monitored.   In  the Arcachon Bay
 in  1980  and 1981,  C.gigas were very badly  affected.  Within
 some  areas,  95-100% of  the two year old  oysters  showed
 deformation of the  shells.   In 1982,  the first effective
 year  of  the  ban,  the incidence of  shell  deformation  was  down
 to  70-80%  and  in 1983  to 45-50%.   £y_en more important has
 been  the  reduction  in  number of  oysters  from  the same areas
 showing  deformities in  both  upper  and  lower shells.  This
was between  70 and  90%  in 1980 and 1981  but zero in  1983.
A similar  recovery  has  occurred  with spatfall; in 1980 and
 1981  there  was none,  but  in  1982 it was  good  and in  1983
 it was excellent  (Vosser  1985).
                              11-22

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  III.  BENEFITS
                                ,»?
        The Agency will perform a  benefits analysis  for  the
  tributyltins during the Special  Review.   The  following information
  summarizes available information on the  usage of tributyltin
  antifoulant paints.

        A.   Antifoulant Paint Use  Pattern

            Antifouling paints containing  TBT or copper  compounds
  are applied primarily to vessel  hulls  to control the growth of
  fouling organisms.   The paints are  also  used  to control fouling
  organisms  on docks, buoys and other marine structures.  Fouling
  is  caused  by the  attachment of marine.organisms to surfaces that
  are submerged or  in contact with fresh or salt water.  Species
.  which  cause fouling include algae,  bacteria,  barnacles, tubeworms,
  hydroids,  and sponges.   These organisms  increase hull  friction
  and weight  which  reduces  speed and  increases  fuel consumption.
  In  addition,  fouling  may  cause deterioration  to the hull coatings,
  possibly resulting  in corrosion.

           Antifouling paints  containing  tributyltin(s) are registered
  for use on  wood,  fiberglass,  aluminum, steel,  and cement hulls.
  These  paints  are  applied  to pleasure crafts,  commercial vessels,
  and  military  ships.   The  U.S. Navy,  the major  domestic user of
  antifouling paints, is  considering  a conversion from cuprous oxide
  to  a combination  of tributyltin compounds and  cuprous oxide in
  its  antifouling paints.   The  Navy has proposed  to use a copolymer
 paint with  very low leach levels to minimize  the the environmental
  loading of  tributyltins.

           The U.S. Navy is  planning to replace  the copper-based
 paints it is currently using on its steelhulled vessels with
 copolymer antifouling paints containing tributyltin and copper
 compounds.  Five to twenty percent of the fleet would be treated
 annually with full  replacement anticipated by  1991  at the earliest.
A maximum annual increase of 100 vessels would  be treated with an
.average of 900 pounds of tributyltin per vessel, constituting an
 additional use of approximately 90,000 pounds of tributyltin
active ingredient per year  (U.S.  Naval Sea Systems  Command 1984).

       B.   Tributyltin Antifoulant Paint Registrations

           As earlier indicated, nine tributyltin compounds are
registered for use in antifouling paints.  The TBT  antifoulant
paints are formulated as single active ingredient formulations,
as multiple TBT formulations, as  any one  of  the TBT's in combi-
nation with copper-based compounds (usually  cuprous oxide),
and  as multiple TBT's plus copper-.based compound formulations.
                               III-l

-------
           There are approximately 340 federally registered anti-
 fouling paints containing tributyltin active ingredients.  Review
 of these registrations indicates that the most frequently registered
 tributyltins are bis( tributyltin^ ^xide (161 registrations),     •'
 tributyltin fluoride (141 registrations), and tributyltin metha-
 crylate (52 registrations).  The 152 single active ingredient
 formulations generally range between 5 and 20 percent active
 ingredient.  In formulations with two or more tributyltin        "
 compounds, active ingredients generally range from 6 to 13
 percent.  In formulations with tributyltins and copper compounds,
 tributyltins range from less than 1 to 20 percent.  The number
 of free association paint formulations as compared to the copolymer
 paint formulations is not known at this time.

       C.  Usage of Tributlytin Antifoulant Paints and Alternatives

           Current annual domestic usage of TBT pesticides for all
 uses including industrial processing water, nonindustrial
 processing water,  wood preservatives, and antifouling paints,
 is estimated at approximately 730,000 to 860,000 pounds of active
 ingredients.  Current domestic usage of tributyltins in antifouling
 paints ranges between 250,000 to 300,000 pounds active ingredient
 annually.   Bis(tributyltin)  oxide,  accounts for 75,000 to 100,000
 pounds USPJ annually,  or approximately one-third the tributyltin
 used  as an antifoulant.   Estimated  annual antifoulant usage is
 100,000 to 125,000  pounds of tributyltin methacrylate, 60,000
 to 70,000  of tributyltin fluoride,  7,500 to 15,000 pounds of
 bis(tributyltin)  adipate, and 5,000  to 10,000 pounds of
 tributyltin acetate.   Tributyltin acrylate and tributyltin
 resinate annual usage  are quite low, on the order of a few
 hundred pounds  each.   There  appears  to be no recent usage of
 bis(tributyltin) dodecyl  succinate,  and bis(tributyltin)  sulfide.

           Many  TBT  antifoulant paints contain copper, or copper
 containing  compounds,  and cuprous oxide is still the main alter-
 native to  tributyltin  in  paint formulations.   Other copper
 compounds,  such as metallic  copper,  are also registered for use.
 Copper compounds are effective against the same antifouling
 organisms  as  the tributyltins;  however,  they tend to be corrosive
 to metal,  especially aluminum.   An  insulative coating is recommended
 on steel hulls  prior to copper-based antifoulant paint application.
 Label  recommendations generally do not recommend copper-containing
 paints  for aluminum boat  hulls.

          Depending upon  the type of paint and  whether there  is
 copper or tributyltin biocides  in the  formulation,  the antifouling
 compound may be  leached from the  paint,  activated by periodical
 scrubbing of the outer paint layer,  or exposed  by gradual erosion
of the paint as  the vessel moves  through the  water (ablative
process).  The  ablative process  is the newest antifouling system
 in which the pesticide is part  of the  paint polymer.   As the
                              III-2

-------
 vessel moves through the water,  the outer paint  layer  is  removed
 and a new layer of  the antifouling paint  is  activated.  The
 advantage of ablative paints is  that they eliminate  the requirement
 for scrubbing the underwater hull  surface.   Tributyltin paints
 are not corrosive to metal  such  as aluminum  and  eliminate the
 need for a protective coating between the metal  and  the antifouling
 paint, except when  copper compounds are  included in  the paint
 formulation.   Tributyltins  also  provide longer protection against
 antifouling organisms than  the more commonly used  cuprous oxide
 compounds,  thus reducing a  ship's  drydock time.

       D.   Benefits  of TBT Usage

           Sufficient data are currently unavailable  to the Agency
 to  determine  the  use patterns and  possible cost  savings attri-
 butable  to  current  and  planned applications  of TBT antifouling
 paints on  commercial,  recreational,  and nonnaval military vessels.

          The U.S.  Navy Environmental Assessment stated that the
 conversion  from nonablative  copper-based  paints  to ablative
 tributyltin paints  will result in  fuel savings,  elimination of
 underwater  hull cleaning, and increased operational  readiness.
 The  U.S. Navy estimated that  use of  tributylin paints would
 result in a 15  percent  savings in  fuel consumption,  amounting
 to an  annual  savings of $150  million once the entire fleet is
 treated with  these  paints.  Eliminating the  need for underwater
 hull cleaning  between maintenance  overhauls  would save an
estimated $5  million annually.  Increased operational readiness,
which  is not  amenable to quantification,  relates to  less  time
 in port, longer range,  and greater speed  (U.S. Naval Sea  Systems
Command 1984).
                              III-3

-------
  IV.   OTHER  REGULATORY CONSIDERATIONS

       A.  Expansion of Special Review

           The Special Review of bis(tributyltin) oxide, bis
  (tributyltin) adipate, bis(tributyltin) dodecenyl succinate,
  bis(tributyltin) sulfide, tributyltin acetate, tributyltin
  acrylate, tributyltin flouride, tributyltin raethacrylate, and
  tributyltin  resinate is being initiated based on acute and
  chronic toxicity to nontarget aquatic organisms.  The focus of
  the review,  at this time, is on the use of these compounds as
  antifoulant  paints.  However, if information evaluated during
  the Special  Review indicates that the use of these compounds on
  other sites  (including but not limited to cooling towers,
  textiles, etc.), results in exposure to nontarget aquatic
 organisms, or that any other criteria are met or exceeded, the
 Special Review may be expanded to include those pesticidal usex
 as well.  Additionally, because most tributyltins pose similar
 toxicity to  nontarget aquatic organisms, and several tributyltin
 compounds, in addition to the nine compounds under Special Review,
 are registered for sites which may result in exposure to nontarget
 aquatic organisms,  the Special Review may be expanded to include
 a much larger subset of the tributyltin compounds than the
 original nine.  These additional chemicals include:

                       Tributyltin Compounds

    Code  No.                              Chemical

    083102                         Bis(tributyltin) salicylate
    083103                         Bis(tributyltin) succinate
    083104                         Bis(tributyltin) sulfosalicylate
    083106                         Tributyltin benzoate
   -083107                         Tributyltin chloride
    083108                         Tributyltin chloride complex of
                                    ethylene oxide condensate of
                                    abietylamine
    083109                         Tributyltin linoleate
    083110                         Tributyltin monopropylene glycol
                                    malate
    083111                         Tributyltin neodecanoate
    083115                         Tributyltin isopropyl succinate
    083118         '                Tributyltin maleate


     At this  time the Special Review  is  based on  chronic  and
acute toxicity to nontarget aquatic organisms; however,  the
Agency is also concerned about the  toxicity  of these  compounds
to humans.
                               IV-1

-------
           The  animal  toxicity data base to assess potential human
 toxicity  is highly deficient for all TBT compounds under review
 (Doherty  Memo  Dec. 11,  1985) although there are some useful
 studies particularly  with tributyltin oxide.  The available
 information (obtained mostly with tributyltin oxide) indicates
 concerns  over  immunotoxicity, teratogenicity, dermal toxicity,
 inhalation toxicity,  and endocine effects.  The Agency cannot
 evaluate  any human risks associated with the use of TBT compounds
 at this time both because the effects are not adequately studied
 and because we have no  reliable estimates of exposure either
 through the food chain  (e.g. oysters) or directly through use
 (e.g. applicators).   Should new information enable the Agency
 to identify a potential human health trigger, the Special Review
 may be expanded to include such risks.

           The Agency  is also concerned that the use of TBT
 pesticidal products may adversely affect one or more endangered
 species.  If information is obtained which supports this concern,
 the Special Review may be expanded to include consideration of
 these hazards.

       B.  Data Call-In Notices

           The.  Agency also intends to issue Data Call-in Notices
 under the  authority of section 3(c)(2)(B)  of FIFRA.   This section
 of FIFRA provides the Administrator with the authority to require
 that  information be provided to the Agency,  which is determined
 necessary  to  support the continued registration of a pesticide
 product.  It also provides the  authority to  set specific timeframes
 for conducting  the required studies and providing the data to the
 Agency.  The Agency may issue  a notice of  intent to suspend
 registrations  of affected products if registrants fail to comply
 wi.th  the requirements of the notices.

           Data  Call-in Notices  are being developed requiring data
 for the  nine tributyltin compounds under Special Review.  Data
 required will  include, at a minimum,  leaching  rate data by
 paint  formulation,  acute and chronic  bioassay  studies,  bio-
 accumulation and biomagnification data,  analytical methodology,
 environmental transport data, environmental  monitoring data,
 and monitoring  of  exposure to workers applying and removing
 paints,  as well  as  data quantifying  TBT  residues in  fish and
 shellfish, and  the  volume  of active  ingredient used  on the various
 sites  for  which  these  products  are  registered.   Additionally,  the
Agency will conduct a  comprehensive  review of  the  data  needed  to
 support  registrations  of other  tributyltin pesticidal  active
 ingredients and may issue  additional  Data Call-In  Notices concerning
other  tributyltin compounds.
                                IV-2

-------
 V.  BIBLIOGRAPHY


 Alzieu, C., et al. 1980..  Evaluation des Risques due a L'emploi
      de Peintures Anti-salissures dan les Zones Conchylicoles.
      Rev des Trav. Inst. Pech Marit. 44:301-49.

 Beaumont, A.R., and Budd, M.D. 1984.  High Mortality of the
      Larvae of the Common Mussel at Low Concentrations of
      Tributyltin.  Marine Pollution Bull. 15:402-05.

 Blunden, S.J.; Hobbs, L.A.;  and Smith, P.J. 1984.   The
      Environmental Chemistry of Organotin Compounds.   In
      Environmental Chemistry, ed. H.J.M. Bowen, pp.  51-77.
      London:Royal Society of Chemistry.

 Brinckman,  F.E. 1983.  Environmental Effects of Organotins.
      Unpublished paper presented at .IVth International Conference
      On Organometallic and Coordination Chemistry  of  Germaniur..,
      Tin,  and Lead;  Aug. 8-12, 1983, Montreal,  Canada.   29 p.

 Chiiamovitch, Y.P.,  and Kuhn, C. 1977.  Behavioural,  Haematological
      and Histological Studies on Acute Toxicity of Bis (tri-n-
      butyltin Oxide  on Salmo gairdneri Richardson  and Tilapia
      rendalli Boulenger.  J« Fish. Biol. 10:575-85.

 Doherty, J.D. 1985.   Memorandum to Linda Vlier, dated December 11,
      1985:   Special  Review of Tributyltin Chemicals-Update on
      Tentative Identification of Potential Toxicity  Problems,
      Request for  Exposure Information and Survey of Tributyltin
      Compounds Registered as Pesticides and in  Toxicology Branch
      Files.   13 p.

 Evans,  D.W.,  and  Laughlin, R.B.,  Jr.  1984.   Accumulation of Bis
      (Tributyltin) Oxide by  the  Mud Crab,  Rhithropanopeus Harrisii.
     Chemosphere  13:213-19.

 Fischer, E.G., et al.  1981.   Technology for Control of  Marine
     Biofouling—A Review.   In Marine  Biodegradation;   An
     Interdisciplinary  Study,  ed.  J.D.  Costlow  and R.C.  Tipper.
     pp. 261-99.  Annapolis,  Maryland:   Naval Institute Press.

 Hall, L.W.,  and Pinkney, A.E.  1984.  Acute  and  Sublethal Effects
     of  Organotin Compounds  on Aquatic  Biota:   an  Interpretative
     Literature Evaluation.   CRC Critical  Reviews  of  Toxicol.
     14:159-209.

Huggett, R.  1985.  Personal  communication with  R.  Hitch, dated
     October 29, 1985.

Laughlin R.B.; Norlund,  K.;  and Linden, 0.  1984.   Long-term
     Effects of Tributyltin Components on the Baltic  Amphiped,
     Gammarus oceanicus.  Mar. Environ. Res. 12:243-71.
                                V-l

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 Laughlin, R. 8., Jr., and French, W.J. 1980.   Comparative  Study
      of the Acute Toxicity of a Homologous Series  of  Trialkyl-
      tins to Larval Shore Crabs, Hemigrapsus  nudus, and
      Lobster, Homarus americanus.  Bull.  Environ.  Contam.
      Toxicol.  25:802-09.

 Laughlin, R.B., Jr.; French, G; and Guard, H.E.   1984.   The
      Physico-chemical and Physiological Bases of Tributyltin
      Uptake by Mussels, Mytilus.  Summary of  unpublished
      paper presented at the Fifty Annual.SETAC Meeting;  Nov.
      4-7, 1984, Arlington, Virginia.  1 p.

 M & T Chemical Co. Jan. 1976.  Acute Toxicity of Tributyltin
      Oxide to Daphnia magna.  Unpublished study.   EPA Accession
      No.  136469.          --...,-

 M & T Chemical Co. Sept. 1976. . Acute Toxicity of  Tri-N-
      Butyltin Oxide to Channel Catfish (Ictalurus  punctatus),
      the  Fresh Water Clam (Elliptic complanatus),  the Common
      Munnichog (Fundulus heteroclitus) and the American  Oyster
      (Crassostrea virginica).  Unpublished study.  EPA Accession
      No.  136470.

 M & T Chemical Co. Oct.  1976.  Acute Toxicity of Tri-N-Butyltin
      Oxide to Bluegill (Lepomis macrochirus). Unpublished
      study.   EPA Accession No. 136471.

 M & T Chemical Co. June 1977.  Toxicitv of Tri-N-Butyltin Oxide
      (TBTO)  to Embryos of Eastern Oysdters (Crassostrea
      virginica).  Unpublished study.   EPA Accession No.  114085.

 M & T Chemical Co.  Aug.  1977.  Letter to Henry Jacoby of EPA
   ;,  Registration Division:   [Data Referring  to the Stability
      of Bis  (Tributyltin)  Oxide in an Aqueous Solution].  EPA
      Accession No. 112780.

 M  &  T Chemical Co. June  1978.  The Toxicity of Bis (Tri-N-
      Butyltin  Oxide (TBTO)  to Rainbow Trout (Salmo gairdneri).
      Unpublished.study.   EPA Accession No.  106966.

Maguire, R.J.  1984.   Butyltin Compounds and Inorganic Tin  in
      Sediments  in Ontario.   Environ.  Sci.  & Technol.  18:291-94.

Maguire, R.J.; Wong,  P.T.S.;  and  Rhamey,  J.S.  1984.   Accumu-
      lation and  Metabolism of Tri-n-butyltin  Cation by a
     Green Alga,  Ankistrodesmus  falcatus.   Can. J. Fish.
     Aguat. Sci.  41:537-40.

Maguire,. R.J. , et  al.  1982.   Occurence of  Organotin Compounds
     in Lakes  and  Rivers.  Environ.  Sci.  &  Technol. 16:698-702.
                             V-2

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 Matthias, C.L., et al. 1985.  A Comprehensive Method of the
      Determination "of Aquatic Butyltin and Butylmethyltin
      Species at Ultra-trace Levels Using Simultaneous Hydridi-
      zation/ Extraction with GC-FPD.  Submitted to Environ.
      Sci. & Techno!.

 Perkins. P.O. 1985.  Letter sent to L. Vlier dated Nov. 8, 1985
      [Findings on Tributyltin Used In Antifouling Paints]. 3 p.

 Seinen, W., et al. 1981.  Short Term Toxicity of Tri-N-
      Butyltinchoride in Rainbow Trout (Salmo gairdneri Richardson)
      Yolk Sac Fry.  Sci. of the Total Environ. 19;155-66.

 Smith,  B.S. 1981.   Male Characteristics on Female Mud Snails
      Caused by Antifouling Bottom Paints.  J. of App. Toxicol.
      1:22-25.

 Thain,  J.E., and Waldock, M.J.  1985.   The Growth of Bivalve Spafc
      Exposed to Organotin Leachates from Antifouling Paints.
      International Council for  the Exploration of the Sea.  E:28.

 Thain,  J.E.  1983.   The Acute Toxicity of bis (Tributyl Tin)  Oxide
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