#3
                                           5PA-S40/9-82-020
                                                  er
                        J
               ana
     Thomas C. £lUvangtf
           of PwticJrie Pf
         Robert K. Hitcn
     Haiird Evaluation Di
         a of ^estts.da ^
U.S. E
1 a 3*
       Aaah-.gts-  - C *

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                                                Foreword

                     Subdivision J  describes study protocols  **iich  may be  used to per-
                form phytotoxieity  testing to support  the registration of  pesticides
                under the Federal Insecticide, Fungicide, and Rodenticide  Act (P1FRA).
                Public comment on subdivision J was accepted  in  a series of  public
                meetings the  last of **sich was held in July,  1982.   Data requirements
                established by 40 CFR Part 158 are discussed  in  Subdivision  J so  that  it
                can be read as a complete package and  so that the.protocols  can be
                be explained  in  their proper context.  ,
J

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                                        SPA 	
                                        October 27,  1982
    PESTICIDE ASSESSMENT GUIDELINES

             SUBDIVISION J

           HAZARD EVALUATION:

            NONTABGET PLANTS
                   by

        Robert w.  Hoist,  Ph.D.
                 and
     Thomas C. Ellwanger, Ph,.o..
     Office of Pesticide  Programs
     Guidelines 'Projects Manager
           Robert K.  Sitch
      Hazard Evaluation Division
     Office of  Pesticide Programs
  U.S. Environmental Protection Agency
Office of Pesticide and Toxic Substances
         Washington, D.C.  20460

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 "~\                     Subdivision J - Hazard Evaluation: Nontarget Plants

 •.  '                  •                  '      Table of Contents


                    DISCUSSION

                       I. Introduction                                .               1
                      II. Organization                                               2
                     III. Major Issues        .                                       3


                    GUIDELINES

                      Series  120:  GENERAL

                         § 120-1  Overview  '        .                                 14
                         I 120-2  Definitions                                        17
                         § 120-3  Basic test standards                               19
                         § 120-4.  General evaluation  and  reporting  requirements     22


                      Series  121:  TARGET AREA  TESTING

                         § 121-1  Target area pJiytotoxicity  testing                 28


                      Series  122:  TIER 1 OF NONTARGET AREA TESTING

&••:';.    4                  § 122-1  Seed germination/seedling  emergence and
*»»"-**                             vegetative vigor  (Tier 1)                        38
                         § 122-2  Growth and reproduction of aquatic plants
                                   {Tier 1)                                         40
                         f 122-30  Acceptable methods and references                42


                      Series  123:  TIER 2 OF NONTARGET AREA TESTING

                         § 123-1  Seed geraination/seedling  emergence and
                                   vegetative vigor  (Tier 2)        •.               48
                         | 123-2  Growth and reproduction of aquatic plants
                                   (Tier 2)                                         49

                      Series  124:  TIER 3. OF NONTAHGET AREA TESTING

                         | 124-1  Terrestrial field testing           ,              51
                         f 124-2  Aquatic field  testing                         •     53

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                                 1

          SUBDIVISION J — HAZARD EVALUATION!  MONTMGET PLANTS
                               DISCUSSION
                            I.  Introduction


      The  performance requirements and testing and reporting
 tores of  pesticide, chemical, environmental, Ind toxiclty p*
 Tin &ftl*r*f+9'+'  **Vk4t  *.-,—..' _*_:	j_ s	  .    .               •  *"»**"•**»£ jrf4.
                      r0n °  m&Ch ***^<*** ***** the Federa
               Ptm9lcide'  •»* 3°denticide Act

 Cod   /?!T^  SerleS*   ThC firSt iS V°luae 40
 Sd informtt'f  f*?*^"* ^^  ^^ specifies the kind oata
 and information  that must be smtmitted.   Section 158.150 specifies

 testSf °XSn.r^UiT"IMt- f°r Ph^otox^^y .(plant protection)
 final              yteRdS ^  rOBUl
     . the second series of documents  [Guideline  Subdivisions  such as
 the present one, published by the National Technical  Int option

 SrS iS? !]  Pr°:iSie ^ tSSt °riteria ^ ^^rting pr^eSes

                                                                     -
ano                                       data recrement     e
SitS i   ^      »t««i»rds and protocols for acceptable testing,
stated with ;a«  m«ch specificity as the current scientific -di.ci
P^« -llo., :«d reporting procedures.  Also provided in Sis
"Mi^"- «• circumstances under which an applicant shoSd
consult with the Agency before initiating a «udy. '
are pri      P*?t*ct**m test «««oool. and reporting procedures
              r*^1^ ^^ wblll««d alon5  with data on env.ron-
              l   TCY *" '"^ to «"•*• '^-  Potential hazard
              on nontarget plants, both terrestrial  and amiatic

                         Cr°PS' "»— «:1..,  «d others Sat «i

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       A parpose common to all tests i. to provide data which will  be
  used to determine the need for  (and support the «ordLg for) pre-
  cautionary ^labeling or other statements^ minimize^ pSinSl
                     *"«**V** *^*'  Generally, the regnant will
         ^^   Precaution«y Deling with respect to nontaget
         such as crop.,, ornamentals, and the like.  However, there
  may be Situations where the Agency will have to develop additional
  precautionary labeling.  For example, the spraying of herbicides
  may 'not be permitted in the vicinity of critical habitats "of
                           II.   Organization
 ad*..    *^          continues with presentation of the major issues
 addressed  by ccmmenter,  with the  publication of the proposed guide-
 lines -  subpart  j,   Hazard Bvaluation:  Nontarget Plants and M
                                              ( " ^ ^948.72978 ,
      The Guidelines portion ;of - this subdivision  (p.  11}  is
         •
                                          ,20
 the subdivision (J 120-2) , basic standard, for testing  (§
 and the general evaluation and reporting procedures^?  120-4).
                1"1 d**la'With' *«?•* «** phytotoxiclty testing,
       is used to evaluate pesticide toxicity to those plants that
 would experience intentional -application . •
 ™.<«          '               series (Series 122, 123, and 1245 com-
 prise  the tier .testing sequences (Tiers 1, 2, and 3, respectively)
 employed to  study and report on pesticide toxicity to nontarget
 HL?   l\  ^ •"•«•.<*•-<*» P*aticid.s are determined through
 a series. of,  test* as. dictated by specific requirements of each
 w««t and tier.   The  tests  are designed to provide guidance for
 gathering pesticide  effects  information on terrestrial and aouatic
 plant  growth and 'development.; The influences  of geographical," sea-
 sonal, and species variation are also  addressed.

     Also contained  in  a section la  Series 122 are detailed proto-
cols for .some of  the-  studies found in  Subdivision J.  At' the end
of each  protocol  are  selected  references  to acceptable methods
that may be used  to develop 'pesticide  .ohyto toxicity data.

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.,. _ j
     Each teat section contains an opening paragraph restating the
circumstance's and for what products, as found in 40 CF.R .Part 15.8,
the data. are. required.  The test sections also contain specific -
test criteria, procedures and reporting formats which, in addition
to the respective general testing information, apply to the accom-
plishment of the studies.

     The execution of studies in the higher tiers depends on the
results of studies in the lower tiers .  The tier system is intended
to reduce repetitive- consultation between the registrant and the
Agency about the need for tests of greater complexity*  As a result,
the time required to develop .data. for registration of a pesticide
should be reduced substantially.              -    • '.    .
                       III.  MAJOR ISStJES

     The Agency received comments from numerous persons or  groups
regarding the :1980 proposed guidelines and the  1982  draft of  this
document.  In many cases the cosaaenters provided information  on
the applicability and the scientific merit of the various . tests.
In response to these public .comments, -the Agency has modified or
clarified all sections and many paragraphs of these  guidelines.
Only the more significant and controversial issues submitted  by
the public are discussed in the following pages.  Many recommen-
dations were adopted by the Agency which do not wax-rant discussion
here. .             .              .               '                   -
               '       A.  General Information .

     Several commenters have expressed  concern  that the  Agency,
through proposed Subpart J and the 'other proposed  subparts,  is
trying to investigate whether all pesticides exhibit subtle  effects
on the environment.  The Agency  is required by  FIFRA to  ascertain
whether a pesticide "...will perform its intended  function with-
out unreasonable adverse effects on  the environment..."  [FIFRA
sec. 3(cH5)].  Th* effects may, indeed, be unreasonable and
unacceptable, even if considered subtle by some observers.  The
purpose of- this and other subdivisions  is  to provide guidance in
the submission of data and other information.   From this combina-
tion of information, an overall  environmental risk assessment
concerning the exposure and effects  of  the pesticide can be  made .
Included in this evaluation is a determination  as  to the possible
effects on endangered and threatened plant species.

     The preamble to the November 3, 1980  proposed Subpart J guide-
lines (FR Vol. 45, page 729495 provided examples as to the possible
uses of the information.  Also,  Subdivision H,  Labeling for Pesti-
cides and Devices j . provides the  guidance concerning various 'types

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  of label limitations ,  precautionary statements  «.. '     .  ^
  relating to phytotoxicity. -.     **..****«***•, or restrictions





                  8*  S^         of Test Data

  -                         ,            prmry    "» «-«•

  -    ~
                     C«   Test Substance.  .

has concluded that

t-aring-ua. products   ^
                        ropra^

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active ingredient in its product if the same data are reaui-ed to
                                                    a
  support the registration of an end-use product that  o
  be produced from the registrant's manufacturing-use products!

  dS«ST!   ? *f ~!T Pr°dUCt " * P*sticide P»duct bearing label
  directions for immediate end-use as a pesticide.)  Section 158.50
  also provides that such data must be suHnitted by an applicant for
  registration of the end-use product, except that the producer of
  the end-use product will generally not have to •ubait or cite data
  ££*" "?„"•!"' to fozffiulate **» «*-»•• product.  This decision
  reflects the Agency's- expectation that manufacturing-use product
  registrants will .b* the major source of registration 'data* .and
  that end-use product formulators will,  in most cases, heed to
  supply much less data.   This decision :: is consistent with the  pro-
  visions of,. and Congressional intent behind,  sec. 3{ c) (2) (D) ,  of  '
  FO.FRA,  which .provides  that:       •           ,

      No applicant' for registration -of >• pesticide who
      proposes  to purchase a  registered, pesticide' from
      another, producer in order to  formulate such  pur-
      chased pesticide into an  end-use product  shall  be
      required  to  -

          •  (i). 'submit or cite- data pertaining  to  the  safety
      of such purchased product? or

            (ii> offer to pay reasonable  compensation other-
      wise required by [f 3(cM1)(BS< of  FZFRA}   for  use of any
      such data.    -                                        J


    ~1JIfPliC:it ** **C 3-(2HD)  ** Congress'   expectation that it
 ™f*   ?* •4"9i«trmBt ot the =«nu£acturing-use product who would
 S£ £ *i9ni*ie«nt aaounts of data pertaining to  the safety of its
 product. (See, e.g., Sen.  Rep. No. 334, 95th Cong.,  1st
 pp . . 8—9 . }         .              •
^»             " **** r^iuirsments were imposed solely on regis-
trants  of     -

              -use  products,  sec.  3{c){2)(0)  might be read to prevent
the Agency  fro*  obtaining data on the grounds that the data pertain
to the. safety of a purchased product.              .         ^^"in


         Teatlin
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                                    6

                 the  active
                                       of the applicant- .


                          ""'
                                                      protection test-

       The  test substance shall be the end-use' product or a
       representative end-use product from the same major
       formulation category :f or that , general use pattern.
       Examples- of major formulation • categories are , we^-
       table 'powders,  emulsifiable .concentrate*, and qranu-
       lars;. ^ df  the; manufacturing-use product is usSSy
       formulated  into end-use .products comprising* two or
      ^ ^30rJ0mal^ioa =^egories,  a  separate study
      f£ \t^fl0rm9& \±th *
      ror each category.)   .   •.
         •hp^d->.aet«» tha* the sutaission  of data using the
          end-use product in question is recommended asl?
 better describe any phytotoxicity associated with  that
      4*   Technj-eal grade vs. fommi;.*-* oroduct   r™™,.r,^

                                               ,.
the representative  .end-use product to .be used in iier 3   -he
of the technic-al  cheaical in .tiers 1  and 2 followfSe intent
and
                                 °f  tectoical Aerial at the Tier t
      ,._ .,    ,    •            -J, at  a  level  that is not phyto-
      to dissolve the material, in water or  other. suitable carrier.


                         _Area ghytotoxicity  Testing.
     1*   Phytotoxieitv .and efficacy teatir^.
     a  confusion between those phytotoxici'ty

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                                 .   7  ,

  Subpart J and those 'normally performed  in
                                                            simul-
  separate the sheets of phytotoxicity and efficacy.  Product
  performance testing and target, area phytotoxicity testing are
  ordinarily and may continue to be conducted simultaneously.


      ..2*' Jf*iirer of target area phytotoxieitv. ; The Agency has
        ^f h     t*Tt "" ^atoxicity ^ta does not need to be
        :SlfCaUSe'^.fe9iStraatS *" ««««"y willing to accept
      overall responsibility of the product respect to efficacy and

  SEST^"^ SeC*  3"(C)(S»-  °— ^ Sidelines Le
  provided to the registrants for those instances where 'data may be
  n.GGQ€scl«
  . B  ,3' ~  w««d-free control plots.   The weed-free or otherwise -pest
  free  control  plots of  proposed §f  120-2(1)' and 121-1{C) E 1) Civ)  wer
                                               the proposed     e
           the maintenance  of  weed-free  and : pest-free plots,   -he
 .conmentars  stated  that  this  is. very difficult, ^practical,  2nd at
 times may be even  detrimental  to  the crops.   Therefore,  the  defini-
 tion of -pest-free" has been changed to  only  reconnnend conLol of
                                                               are
                         P°r exaffl^1«' ^  «ntrol process  of  weeds
                                  vfeh* la^l-  As  stated  in  sec.
         f Fina> a pesticide may be applied "...employing a^y

        f 5tP  cmtl°'15 S0t ^ohiiited * the labeling..;-  m the
 proposed Sttbpart J guidelines [proposed § 163. 121-1 (c) (3 )] ,  all
                                                            ,
    ew            ; prohibited by the label would have been eval-
 uated with.-resp.ct to pesticide application and movement in the
                       counters have stated that testing all
            "•«**• ,S2t Prohibited by the label is impractical and
                                                               . r
               T* u  i3-disco^a^ because of the celic.t^ "   *
               of  that situation.   Therefore,  uSe of some methods
of application which ^are  found on the label, need only,, bl tested?
If ^a -worst  case- application method can be readily • determined
prior to testing, then testing, may' be limited to that case.  Sup-
port for the use of -.that  method should be famished to the -Agency.

                                                          con.at.rs
       .                                      synergism)  and serial
     cations tests were excessive  [§  12l-1(b)C5)  ^and  (6)].  -he

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ea                          °ce  «-1  °^*nuary 1982 has
eliminated, in most cases, the r
  ea                                                       as
  emnated, in most cases,  the  requirement  to  «uhi.4<.
                 the treated trees*



      The reporting .of general population shifts  in pastures' and
    .
     7*   Subaecment Planting (rotational frn^^   Comm.ni..^.
sion J [f .121-(c}
                      are  used ",tq evaluate, the phytotoxic
                  ^ontart?et ^e* Phvtotoxieity' Testing



              regmirenen^ for' non

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o
or
       The Ag«acy is retaining Subdivision J nontarget area phytotoxi-
  £  , ^f,     th°Se SitU"ions **« ««» information is leSed!
          tlsf ^ f fideS,a "* °f' Standards «* reporting, formats
           !          ta Vfhen th*y •" «<3uest**'  several examples
             ^ *?** f6^*6* •*•«   <1>  »«*«*• Posed to endangered
           ened pxants listed -by the  United States Departmen- of
  Interior,  Fish and 'Wildlife Service;  (2)' initiation of a rLuttable
  presumption  against  registration (RPAR)  where a phytotoxicity
  problem may  exist; and (3)  where a  specific phytotoxicity problem
  arises  when  general - open literature data are not available.

      : The Agency will inf orm the registrant of the, chemical in ques-
  tion, concerning the  phytotoxicity problem and the specific data
  required to  address  the problems

       2*  Terrestrial  species  selection.   a the proposed Tiers 1  and
  2, seed  germination/ seedling  emergence and vegetative vigor tests
  (proposed ;§,§ -.163.122-1  and  163.123-1), ten specific  kind, of plants
  were  to , be tasted.  This made  the -guidelines 'somewhat - inflexible  and
  did not  readily .'-permit  the  use  of auch screening test data already
  generated by companies.  The  selection now states. that soybeans,
  command a dicot :root  crop are  to  be' tested, and that seven other
 test species :are .to be  a balance  of monocots  and dicots.   Corn and
 soybean were . retmmfed, due to  their  economic significance  and the'
 quantity of - .pesticide -research performed  using  these  species.   By
 increasing tliis flexibility of species selection, tests  that are
 normally performed by the developer/registrant  during screening and
 initial field/testing may •• often be used.   This  change  will result
 in -a significant: cost reduction for this  test.
 ,   ,  3*   agaatic species selection,  several ccaraenters noted that
 inclusion of :five aquatic species at the Tier 1 and 2 level can lead
 to  expensive and unnecessary testing.  They suggested that only one
.species,  probably Selenastrum eapficAmuMa. be tested at the Tier
 1 level.      '        '

      After careful consideration, the Agency decided that this
 species selection was indeed unnecessary and that the. selection
 could be  based  on use pattern. •  Selenastrun will be tested for all
 terrestrial  or  aquatic outdoor  uses,  if an outdoor aquatic use
 pattern is anticipated, the other four ao^atic species would also
 be.  used.

      The  aquatic  species selection was based on those species -hat
have  been, extensively tested and for which the growth paramete-s
nave  been  strictly -determined and specific strains are read-'ly"
available.   For .these reasons Lestna gibba G3 is chosen over' Lemna
3anor and  Selenastrua capricorrmtua over Chlorella vulgaris .   The
diatoms are  used  because they have been shown to be very sensitive
to water pollutants,   toabaena floS-3Cuae is chosen as a reoresen-
tative of  a  group of  plants  that can fix atmospheric nitrog'en.

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                                    10
        The Coverall selection was made to obtain a broad representation
   of  aquatic  plants and provide  some  insight into variations  of  "
        rr                                  "*  Plant     es  r
     - Tier 3  ( dicots , monocots ,  ferns,  etc ..)  addresses the  fact lhat
  .plants other  than .alga*  inhabit  acetic  areas,  Again  thislest  is
  to note the variation of .effects U.e.,  tolerance  or .resistance)
  •to- the pesticide.      •   '    . '       .,            ;  .  -  .   <"lce/

       4>  P°f*gea or application' level,..  Many •eonmenters .to  the
  proposed Sufapart j guidelines stated that  three tiaes  the label  rate
  was an. unrealistic quantity to be assessed for aontarget area
  phytotoxicity,  This statement was based on  information  from actual
  uses and .exposures.  In  response to, .these  comments, the maximum
  dosage or application level was  set at the maximum label rate.
  Again comments were received that this rate was excessive and that
  the rate should be based on environmental exposure.
     ..  It was not the. intention of the Agency to 'perform these tests
  after environmental exposures had been determined or modeled,  if the
  registrant, however,  decides to perform these tier tests after deter-
  .mmation of the .environmental exposure, then a rate equal to at least
  three tome* the exposure as found in the adjacent nontarget area, may
  be  used.  It aust.be  remembered "that the adjacent nontarget area can
  be  the adjacent desirable plant of another species 0-1 meter or 100
  meters distant.,  therefore, the use of this exposure level must be
  supported with' appropriate data.

       On the other hand  the use  of the maximum label or environment -
  exposure rate does not  preclude the voluntary testing and submission
  of -phytotoxicity data where the tests were performed using higher
  rates,   it is noted that dosages 'used during manufacturing screening
  tests  would -have a greater ^tenden'cy to . exceed' this required dosage
 or application level, and would thereby . increase  the probability of
 acceptance of -these screening ' tests .   .              :
                           Mutagenicity Testing.

      Since proposing the' concept of a plant mutagenicity testing
•scheme .in Subpart J, many - registrants.and other researchers  have
 expressed concern-that these tests would not provide meaningful  data,
 AJ.SO, no incidence of plant autagenicity has been substantiated  for
_target.area crops, or nontarget area •plants. '

      Several commenters suggested that this set of tests  undergo an
 extensive series of evaluations before this type of testing  be in-
 cluded in any finalized ruling.  Also, commenters and others pro-
vided references which question the-validity of using plant muta-
 genicity studies to evaluate huaan mutagenicity.

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                                   11
  withdS thl             h"e COfflMntS' the Agency-has decked to
  withdraw the requirement for the plant mutagenicity studies
  extensive testing can!,* performed to show the aorl
  usefulness f or ^ this requirement.
                       G*   -ier 3 •Fiel<* Studies.

       Several commenters  noted confusion in the  requirements of and
  tne  differences  between  the  Tier  3  aquatic and  terrestrial  field
  studies  and the  Tier  4 geographical and seasonal field 'tests.   This
  confusion was -generated  by the tier progression statements  where one
  processed  from  Tier  2 to either  Tier 3 or 4, depending  upon a
  plex  set of progression  requirements.    '
  «. ^*               conflision' ^1 field. studies were  combined
 at the Tier 3 level with respect to either terrestrial field  or  '
 aquatic field testing.  Geographical' or seasonal considerations
 are Deluded in the Tier 3 tests.  There is no longer any Tier 4
 testing.                                    ,
                                 fixation Studies.
      All testing,, of, microorganisms was removed from Subdivision j
 except for testing of alga*.  Therefore, testing. of the nitrogen
 fixation potential as affected by pesticides was removed from  -
 ^SV«SK!?'f*.  Thl* study'wi11 *• considered for inclusion in pro-
 rS6d f"1^-"- « ^«H»f «ith pesticide-microorganism effects!
 Comments received will b, used in the development of these require-
 ments when this  subdivision is prepared.                   -eqmre-
                         !•   Sorption Study.
    ^The  requirement  for  a  sorption study as proposed in Subpart 3
was based on  a  theory of  possible  mode  of exposure of aquat
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                           12

                  J.  Spray .Drift si^-i —.'
 tlon series from SubdiJSLn                ren°Ved ^ sec
                        J and
                   K«.  ?ier Progression.

 1 ^            9eneral *^eed -that the- BC10 vmlu. for the Ti«r

the .testa must be repeated '  *rr™tal effect of 25 .percent, then
sits:                               "--s
    Commenters also stated that
      »., not
          eneral agreement does, not exist among researchera
                                              "

     S^S=SS~=3
                        or EC95




        2 levels, basina nia-*.- -i^-i	^ __
                                  3n use patt

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                                 13

     - The TM-jfiTmi-m dose. level has been reduced to the maximum label
       rate or to 3 times the maximum expected environmental exposure •
                      L<  Statistical Analysis

     Several coasmenters stated that for the results to be statis-
tically significant more1 replicates and/or 'a greater population
size would be required.  A basic part of scientific analyses is to
have sufficiently. large populations in order that the results be
meaningful.  The Agency is making the selection of population size
flexible as each study would require a different number  of  indivi-
duals.  It should be noted that each species has a different seed
germination and survivability rate which has a. direct bearing on
the statistical significance of the results.  The Agency encourages
the use of the largest possible populations for each of  the tests -
in order to approach the 90 to 95% level of confidence .with a
significance level of less than 0.10.  The following references
are provided concerning sample size selection.  '

Casagrande, . J.T., Pike, M.C. , and Smith, P.G. 1978.  An  improved
  approximate .formula for calculating sample sizes for comparing
  two binomial distributions.  Biometries 34s 483-486 «~

Fleiss, J.L. 1973.  Statistical; Methods for ........ Rates, and Proper-: ••-.'
  tiona .  John -Wiley and Sons, Inc. New York.

Snedecor, G.W., and Cochran, W.G. 1967, Statistical Methods , . 6th Ed.
  Iowa State Univ. Press* Ames, Iowa.

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                                   14





          SUBDIVISION J — HA2ASD EVALUATION:  aONTABSST PLANTS




                              GUIDELINES
 Series 12.0:  GENERAL






 §-"•1-20-1 ' Overview.


™
runoff, .soil • erosion, : spr.y . drift/ etc. ) .
     (A)   Toxi=tty to pl«« in the t«5«t .r.a (j ,21-1),
                                                   '
                                                   •      '•   -

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                                   15
  registration of each . end-use product with the prescribed use pattern
  and each manufacturing-use product .used '-to make such an end-use
  product.     •:•             -   •      ..._.'•

       (2)  The -test substance- paragraphs state the kind. of pesti-
  cide material' that Bust be used 'in each test.  The test substance
  for studies in this subdivision may be- the- technical trade chemical,
  or a representative end-use product.  Generally, each of these
  test substances is prepared by the basic manufacturer of a- pesticide
  chemical.                ,             .
       (c)   ires*"*g to meet requirements.  -Since studies found in this
  Subdivision would ordinarily be conducted by the basic manufacturer,
  pesticide  formulators would not often be : expected' to conduct such
  tests themselves  to  develop data to support their individual prod-
  ucts.   (See 40  CFR § 158.50 concerning -the formulators'' exemption. )  .
  They may do so  if they wish,  but they may -also merely rely on the
  data, already developed by the basic, pesticide 'manufacturer.

      (d)   "^gg-et  "6* phytotoxicity testing waiver of rfemii^m^^ .
  (1) The Administrator has determined that  efficacy test, data include
  target area phytotoxieity testing data,  and that data ^submittal for
  such testing may  be  waived, by his.  authority under FIFRA Sec.' 3(e)(5)'
  for most kinds of pesticide products.  (See 44 FR 27938-27940,  F-iday'
 May 11, -1979-. J  Such products generally -include all pesticides  whose
 uses result  in direct or  indirect application to plants  in the  target
 area such .as  agricultural,  lawn,  and garden usa.
      (2)  ^^ though the" Administrator will ordinarily waive  the
, requirement for submittal of target area phytotoxicity test  data as
 indicated in paragraph (b)(1) of this section, he reserves the
 authority to require, such data on a case-by-case basis whenever the
 Administrator deems that such data are necessary to evaluate the
 acceptability of a product for registration,  if it is determined
 that data phytotoxicity for a .pesticide -are -necessary, the Agency
 will promulgate the. specific target area phytotoxicity data  require
 ments by letter to a specific registrant or by general notice...

      (3)  Thus, the guidelines in this subdivision should be used
 by registration applicants  as phytotoxicity teat standards and
 phytotoxicity data reporting requirements when target4 area phyto-
 toxicity data are  submitted to support registration applications.
'The guidelines may also be  used to provide guidance on testing
 to support the claims  and directions for use on product labeling
 for products for which target -area phytotoxicitv data submittal is
waived.                                         "         •
men1
 {8)  Sontar?«t **•»« ohytotoxieity  testing.   (1)   Data require-
s.   Data concerning the determination  of  outdoor  pesticidal

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                                     16


              or
        (2)   Testing aeh*^ .  Tests  to the  lower  tiers  n  »«* ^
                                                               -
   wfR part  158.  Where  data  are  stiimitted to fulfil  *-K^
ments of one subdivision  ero«« ^.J^     " rui£i11 the require-
a*d. by 'the reglsS^t !«  ?£ ! «*«r«nce« to that data should be
      y »• .egistrant if  the .data ^e also required elsewhere.

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                                                      17

                                  registration applicant is referred to Subdivision

                      120-2  Definitions

.
                       *n
                      ' (•)  The term "SDx" means that  ^-ite— ml
                     dosage required to d.«rla.atmlly 5"^ pi

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                                   13


                            50 percent
       Ch)  • Th*  titan ' "nontara»t


 or
      Ci5  The tern "peat-free"
 possible.
                                       as free of pests
         are
          can
                                             °
                                           l
                                            * action detrimental

and

                                                °f
                                                      "
          thaty *.
response My occur during germinaion    oh  di^              c
maturation of plants, and m*v^~^^' ^±*f.««**i»tion,  and

sp,cl.,.
                                         desirable and imdesirab!.

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                            19


    12(?~3  i*sic teat• standards.

                          •                       -
                 pr«c«ae.,=« ln the p.rjom.,,ce of «at particular

 Si*
 s-i*-*"^ = - =£S

    § 62-2.   ;              imits, if any, certified in accordance
used           ;l




       their purlST?.? S!°ref Mder conditions that

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                                    20
                                      to possess
                                  «*  che^cal properties of the test
                levels u*«5iVin the study, it should  not  produce
                or toxic effects to' plants.    -r,.        P««««

       (S)  Where 'the test substance does not readily-
       (i)   An -analytically pure_.. grade 'of an active; ingredient;

       (11)  The  technical  grade  of  an active . ingredient;

       (iii) An inert ingredient  of  a  pesticide  formulation,


       !iV>  A C0ntaminant  or ^«rity of  an  active  or inert ingre



                                                of  » active or
      (vi)   The pesticide, formulation;
sections- of this subdivision:  121,  122,  123,  and 124

   -  (2)  Healthy plants aast  be  used.    '

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                                   21
       (S)  The population size  of  m&ch replicate or treatment should
  be large enough to assure meaningful results.  Sample
                    (section  series 122 and 123) should
                                         2S or 50%
                                                        £ ield
                                                  -

 system of random assignment  of  the  test plants
 group's is- required.   "
                                              -
                                     ss: s •
in=lui?«  5?1tPMgt'   (1)  M1 *5^P^t used in • .conducting the
xncluding ec^ip^nt used to prepare and administer the tesl aS
*,•  ^(2J  Th" aPPUc*tion equipment used in testing produc-s' in sma
field plot ^studies should be designed ro simulate convenSnal *S

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                                    22

  small plot spray.« can b« identical
  ccdaidcial «•«•••»«•—-»	.
          should produce results
      t. on
   cial

                                                                  on
                                                                plot
                                                              au
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                                  21'-*

f« .,    r**  ^"^i^isa «nd composition of any>ehicles
(e.g.,  diluents,  suspending agents, . and -emulsifiers ) or o2er

          used 'in th. testing of -th, 'substance.
                                of »*i- reporting requirement
                                  to Sabdi^ision o (f 61-1, §
     (S)  Ontoeated  eostrol  ( cheek 1  olots.   Detail.^ ^M<- 4*4   •

                    used  as  controls fo'
             et     T*"* tO ^^ ?«sti=id«  °^ cneMcal
             etc.) :and cultural practices,                   .  .










                                       "™™

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                            24
     CD)  Turbidity (visual),  conductivity (if possible  . „
dxssolved o*yg« (for submerged plants only);^d      '  ***







                      --
                                .               .  „.
^-11™^-

term st«S                   "   C°nditi°M -^ ^ «*d for lon
                              C°nditi°M -^ ^ «*d for long
                                  for th* duration of fi9

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                    "* *** iatervml fr*1"-  *PPlications should be
                 products are applied. is  a  tank
  wi                                           , •
  with identification and formulation  for each product.

       (ill)  Timing of applic^i^..   when toe test substance,
  particularly a  herbicide, plant regulator, desiccant,- or defoliant
  is applied  to any desirable nontarget plaats within or adjacent^o'

               rsf: piant>s sta9e °f «rowth °r *™u*"Jt..f
              should be described .in test reports. ••       •

             Aerial  applications .  In. addition to the detrimental
                                times o£ *PPli=*tion (or

                                   each product °r
           Observations.  (1).. Observation* should b. reported to  '

              ^^ions, either inhibitory or, stimulatory,  between the

              organisms, and the untreated controi teat organisms.

                            °tOXiC S*mpt0mS {c^°«»^' necrosis, and
      (2)   Observations  should be  reported  in  sufficient  detail  as to


          Pt"ali011 °f  th«-rM»lt-
                                                           ,  to be
                            ,  should  include  the. degree or  extent
r.^JJi^"?  dlltfla"ntal  or «»'•»• effects to be considered and
reported during the observation period of terrestrial studies include


      Ci5.   Stand or plant  population,-    ;
di.«      1°V*r*11 T±9°r of th8 -Pl«« expressed as height, weight,
diameter, length, or other similar aspect of -growth,-
malfoi^il Fhy*0t°xicity or visii^ symptoms such as discoloration,
malformation, desiccation, or defoliation;


     (iv)  Lodging of plants;



           Effect on root growth and structure;
                                                          to

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                                  26

  sr.S                       S5



  .„,,   (*1, AV*Mt two »«tliods of  evaluation (such as quantitative
  and qualitative determinations)  should be used in the S^tton If
  ^stxcide effects on growth- reproduction, and yielTof J^ts L
  greenhouse and controlled chamber  experiments,  ^hen direct measure-
  ments cannot, be made, *uc* as in large field evaluations ,  a zero
  to one hundred (0-100). or^zero to  ten (0>10) rating scate

                                                  "
                                                             u
 S •ST(',n"J'r0 
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                               -  .27%..

     (2)  Statistical analysis is also useful in evaluation of
interactions resulting from studies  supporting tank mixtures or
serial application*  [See  121-Kb) (5-) aad  (6)].   '

     (e)  References.- Copies of references or literature used in
modifying the test protocol, performing the test» making and inter-
preting observations, and compiling  and evaluating the results
should be submitted.  Copies of unpublished literature should also
be included.  Copies of the recommended literature referenced in
these- guidelines are not required.        •

     (f)'  Special test requirements.  In  addition to'the data
required in this subdivision, data from other tests may be required
by the Agency for-making  judgments regarding safety to nontarget
plants*  Such data will be required  where there are special prob-
lems, such as a proposed pattern of  use,  mode of phytotoxic action,
or a unique chemical property.  Methods are -usually derived from
those already described or cited in  other subdivisions of these
guidelines-.                                   .           •

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                                    28


   Series  121:   TARGET AREA TESTING
  §  121"1  Target
                  area
                        phytotoxicity  testing.
  generally will be waived by 40 CFR Part 158 to 3       the

                end"USe Pr°dUCt lBtMd«1 for outdoofS
               or outdoor planting of treated material  [

               T^0'11' n°ted in § 12°-'^5' the Agency   y re
                data fro* studies provided for in this section
  fiil.                               this section need not be fui-

  fon^ol         ?ideS WhiCh Provide long-term or total vegetation
  control,  e.g.,  clean yard chemicals,  desiccants and defoliants.


  also  12)H  Experimental use permits.   The registration applicant is
  also  reminded that  an experimental  use  permit may be


  °                                                '
                                                               e
                for  information  concerning  experimental  use  permits
            **
      (1)   Test substance.  The test substance shall be the
?T,r * repr"«tativ«  «*-»•• Product  from theam
formulation category for  that general use  pattern.   Examples  of
maor formula
                                       use pattern.  Example
       formulation categories are: wettable powders, emul.i-
 concentrates,  and granulars .  (if the manuffcturing-u^


 "               **         "
                                 —       -	c"~^™»*«^ &WQ O2T uiw^ c ui

                 -      -  -  separate study  must be performed with a
        end-use  product  for each category.)



      (2)  Test species.  Those  desirable  target area or pest host

Plant species as listed  on  the  label  (for example,  the crop Pl^t or

ornamental) which will be within the  target  area ihould bTtes'd

:he plant cultivars to be tested should include representatives of
the cultivars that are most likely to  be  used.



     (3)  APPUcation3 levels.   (i)  The  minimum, maximum (or -he

greatest allowable concentration),  and  2  times  the  maximum libel

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                                                     .29

                    application level or rate should be tested.  Levels greater than

                                effect (or no-effect} level should alse'b* determined.

                               The multiples of the, application rate to be tested are
                               .us quantities of the,formulation in the label-recommended
                          ty of carrier (such as water), to. be used per land or aquatic


                                          Products .with labeling which allows or recom-
                           H            -           '                            r rec
                          the addition, of separately-packaged adjuvants to the spray
                    tank ;should be supported with data indicating any detrimental
                    effects- (s«ch as -increased crop phytotoxieity)  which may result
                    from their addition to the. pesticide,  especially a herbicide,
                    Plant, regulator, :d*siccant, -or defoliant.   If a range of adjuvant
                    rates is' recomended ,  the maximum rates  within that range should
                    be evaluated in  conjunction with the intended pesticide product.

                         (5)   Tank mixtures.   When tank mixtures  are reccamended on
                    product  labeling, a  study may be required  on'  a case-by-case' bas^s
                    to demonstrate the  extent of  antagonism  and .synergism with resoect
                    to detrimental effects  on nontarget plants by, -.the products' of tank
                    mixtures.  Antagonism  and synergism are  best  evaluated in adjacent
                   plots, where possible interactions  are  sub jected.to* statistical  •
                    analysis.  See | 164-4  of Subdivision  N  for 'possible- combined test-
                       --                   •
                     't C!,}; s«»-^-*l agglieatiQRg.  Data requirements for serial  appli-
                   catioa(s) at one or more pesticide(s) preceding or following  '
                                     T ** saae crof>
                   are identical to those described in, paragraph .( b) ( S ) of this
                   section for tank mixes with respect to phyto toxic ity, when such
                   serial applications are reccnmended on, the label.  See §  164-4 of
                   Subdivision N for possible combined .testing, .   . .

                        {73   Ii£«-   Th« test should be performed -in .greenhouses or
                   wherever  the product is intended to be used.

                   ' .    (8)   Protocol.   The protocols, methods, or practices should be
                   S^LTP f^r ^ ^icipated registered use of the pesticide
                   product.   Specific points of information that -should be addressed
                   concerning use patterns,  application methodology,- cultural prac-
                   ^T*!;  r*8P°nSM'  ^^ subsequent planring are -found in paragraph
                   (c) of  this section.                           :

                   .     ^*}~   JSF3^^?'   In  addition to the information required by
                   J_ 120-4,  the  test  report  should  include the following information
                  7ltVKr!fP*Cl:  to Ph^otoxicity to the plants within the target area
                   (with the  exception of  weeds).   This information should include
                  the method of application,  cultural practices,  plant responses ,
                  subsequent plantings,  and use patterns  that may be involved.
J

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                                   30

       (1)  S«aeralinformation.   (i)  Timing of.
               f any pesticide-, their stage of growth or
  at application should be described in the test report!

  „.   (ii) •M*t^relogic«l conditions.  Where .meteorological condi-
  tions cause detrimental effects on plants which in tarn allow the
  pesticide to further adversely affect the plants, ' the specific
  £S!±i'" *Jf J! t such as soil moisture content and
  temperature, which. are directly affected by meteorological con-
  ditionp,  should also be reported.   Soil moisture may be observed
  azid^expr eased .in terms of • dry and  cracked,  waterlogged, or other
  similar conditions.   Organic matter content of  the  soil should
  also.be reported.-'                             ' •

   4J.   fiii3 ggray  Dilutions,   .in foliar applications,  when a pesti-
  cide  is applied as a  diluted spray and the  quantity is  dependent
  upon  the  number of trees  per area  or ^density of 'vegetation,  the
  total spray  .volume per  .unit  area,  and  the concentration of. the
  applied pesticide  should  be  reported.  '.'  :
            gntreated controls  f cheeks).  ,la phytotoxicity evalua-
 tions, all treated plots, plants, and/ or ' commodities should .be
 compared directly to untreated control plots, plants, or commod-
 ities.  All quality and/or yield evaluations of pesticide-treated
 plants or ceamodities should be compared to control, plants or
 commodities receiving the same pesticides (e.g., herbicides,
 insecticides, 'fungicides) except the one being evaluated.  Detailed
 descriptions of plots and' plants used as control treatments for
 comparisons of detrimental side effects should be included for
 each test.   Since such control 'plots are established to evaluate
 any direct  detrimental effects of the pesticide on the crop or
 commodity rather than to evaluate efficacy,  any detrimental
 ef .ects on  the crop or commodity resulting from pests should be
 controlled,  in other words,  the control plots should be both
 untreated by the pesticide in question and as pest-free as reason-
 ably possible.  If,, in addition to the untreated control plots
 plants,  and/or commodities,  a 'registered product is applied (as a
 standard) for ccaparison' of  detrimental' effects,  data should
 indicate  the -standard product ' a ' name',  active ingredient,  dosage
 rate,  and phytotoxicity results.'  Where  infestations of weeds
 occur  in  check (or  test)  plot«,.th«  degree of infestation and
 species of  weed(s)  should be  reported.

      (2)  Ose  patterns.   When the  following  use ' patterns  are  found
on the label,  the corresponding information  as detailed below should
oe reported.                       '     •

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                                               31

                             in field • crops*  Effects of pesticides on desirable
                     area .plants should be evaluated and reported*  The extent
              and duration of the effect should be expressed in terms of stand
              and..vigor,_recovery, yields, and degree of .phytotoxicity.

                  ;.lii)  Use on pastures and rangelands.  Effects of pesticides
              on desirable target area plants should be evaluated and reported.
              Severity and duration of 'adverse effects on desirable plant species,
              expressed in terms of stand and vigor reductions,'recovery, and
              changes in yields, should be reported*  Data should be submitted
              addressing reseeding intervals which minimize adverse .effects on
              rcseeded plants, and animal grazing reeemaendatid'na. which allow
              recovery of desired plant species.  If the applied pesticide kills
              all vegetation In the treated area for. an extended period'.of time
              resulting in bare spots, 'the registrant should record the duration
              of this effect, estimated /soil 'loss by erosion and any changes in
              vegetation cover (desirable or undesirable)*

                   (iii)'Pse on and around fruit.and nut trees.  Applications of
              pesticides on and around fruit and nut trees require evaluation and
              reporting,of detrimental effects-on foliage, and changes in, growth
              compared to preapplication measurements and simultaneous controls.
              Pesticide applications to bearing fruit and nut tree areas also
              require evaluation and reporting of detrimental effects on yields
              and commodity .(produce) quality for the year of and the year after
              application.  Supporting data should address, for all trees, the age
              of the trees, the transplant-to-application interval, and the maxi-
              mum allowable extent of contact between the' pesticide (with par-
              ticular reference to herbicide'.spray drift) and trees.  For ground
              sprays, unless the .pesticide is broadcast over the entire orchard
              floor, data should indicate the application technique .(band, spot,
              shielded, or directed spray application) and the size of  the
              treated ground area .around the tree trunk*  Assessment of root
              sucker - treatments should be made where applicable.  For foliar
              sprays, the data should include the volume of finished spray applied
              per unit of land area; concentration of product in the spray solu-
              tion, and the extent of foliage,coverage (such as volume  of finished
              spray per tree or application to the point of runoff).

                   (iv). , Ose- on.-:lawna, and turf.  Evaluation of'effects  of pesti-
              cides on representative species - or,cultivarS of desirable  lawn  and
              turf plants should include, such factors as color, density, percent
             .cover, growth rate, •,rooting, and tillering.  If use on bentgrass
              is intended, this highly susceptible species should be evaluated.
              Data should address- use--on newly-seeded lawns by demonstrating
              safety to representative species and cultivars of desirable  lawn
              plants to b« named on the label-as kinds on which the, -product  is
              safe to use, with seeding-to-application intervals-(if appropriate).
              Data should also,.address-.-use of an appropriate application-to-
              reseeding interval for each of these desirable lawn plants that
              may be reseeded.  Interactions between herbicide  application  and
J

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                                  32

 lawn cultural practices•(such as raking,  mowing,  mowing height/
 watering,  and fertilizing)  should be evaluated for possible
 adverse effects on'desirable lawn species.   In situations where
 fertilizer and a pesticide .are applied serially and both types
 of  products may contact -the  emerged crop  foliage (such as in turf
 or  lawns),  the interval between application of the pesticide and
 the fertilizer should be  reported, as well  as any resultant phyto-
 toxic effect, stunting, or  discoloration, and recovery time for
 the injured desirable species.

      (v)   Use around ornamentals.  Phytotoxicity data"in support of
 use on or  around an ornamental should include an evaluation of the
 sensitivity of representative cultivars of  that species.  Since it
 has been documented that  cultivars and varieties of the same species
 vary in their susceptibility,to 'injury, the limited nature of test-
 ing should be addressed in product labeling.  Test data should iden-
 tify the method of application as to"'1 directed spray and/or topical
 applications.  Growth stage  of the ornamentals and the transplant-
 -to-application interval  (when applicable)  should be indicated
 in  ~.ne test data.   Information should be  submitted on specialized
 nur -«ry cultural practices  employed in tests, -such as use of
 arr-ficial.soils,  mulches,,containerized  stock, and other pesti-
 cides.                   ;  •,                        '

      (vi)   Use in  forest  management.1  The effects of the pesticide
 on  desirable plant species commonly present in forest management,
 in  addition to the desirable forest trees,  should be indicated in
 the'report with any detrimental or adverse  effects "that the pesti-
 cide, may cause.  Special  attention should be given to pesticidal
 effects on  noncompetitive ground cover species that-aid in the
 land management practices such as erosion control.  Appropriate
 testing and assessment techniques adapted to the size of the plot
 should be used to  determine  the effect of pesticides on all plants.
 (A'  recommended reference • is:  Phillips, E.A.  1959.  Method's of
 Vegetation  Study.   Holt,.Rhinehart, and Winston,' Inc.:  'New York,
 N.Y.   107 pp.)                ,    -          ' : - ;     ;

      (35   Application methodology.  All methods o'f'pesticide appli-
 cation specified on the -label should be evaluated and -reported.
 Specific detail as to descriptions of equipment design, adjustment,.
 and operation should be :provided in test  reports involving aerial
 applications 'and applications-using 'conventional -farm equipment
_(such as tillage or planting equipment),  irrigation systems,
 mechanical  incorporatiori, directed sprays,  mist "blower  (air
 blast,  air  carrier), subsurface1-placement,  or band rather than
 broadcast  distribution. .  .... ;   ,':•:,,              :

      (i)    Aerial  application.  Guidance  and the data requirements
 for 'testing aerial applications,,will be provided in a subdivision on
 spray drift exposure assessment.

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J
                                                   33

                             Irrigation system
                  fi
                  field should be  examined  and the
                  oh
                  of  hax^estafale  ccanodity)  as an indication of pestic
                  Data from such  plots should ;b. reported for eaS S
                     -                             f                '

                                      **** Spriakler '*•«•*»» overlap.  On a e«nt.r-
                                 w     t0 ^ 38Verml "Pie" «««10B.-for treatment
                 subplots m one half with the second half -as the control.   The
                 concentration' of. -active ingredient at .several .no«le8 alon/?he
                 lateral should also be determined and reported-.  ,

                      CC)   For .surface- irrigation  systems  such 'as flood,  furrow  dr
                 and surge,  the  following data should be submitted.   ConcenSatlo^
                                      !

                                                                                    ;


applications.  Guidance and the data
             blast «- *^ carriers^
                         ,                                              ers  w
                provided in , wbdivUlon on spray drift exposure assessment
               t«m, volume  of  liquid' or  gas  applied p   unit aref^or gen«
               broadcast  applications - or  linear  row  dfst«ce  for band aid row

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                           34 •   •



   .^iSLT^IbHs'S^^*^^

   '^'^rssisx'tsr,"^to '*y °- *«*«•
   h.li=opt=r, or fi,.d-wL, ScrS° ' P  P1°9 
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                                    35
                                                  «op
  prior to or ju,e .rt.r a p.sticia. application ««y, by
           d.sira.,1. ,!„„ or by d^Lia, 5ro«h
  practices should be reported la teat results.




  _   (5)   Tafget area PlaBt '••"""*;.-   The ..detrimental effects on
  crops,  coonodaties .(produce)., -or any ..otker^sirable ^plmnt s-pecies

  TLC?TT   f Within ^ target.ar.ea.sho^^.^eval.oat.d a^d     "
  Th.  following are some of .the characteristics - that should be



       {1)    Stand,   crop stand counts, reported  as  percentage of


  SSrif irStr°1 "^ St*ndS'  •he«id"b« suinitted to support pesti-
  cides.- applied prior to crop -emergence.      -          -.*'*-   ±«»*.A



      ^ij , -Zi22E'"" C^OP vigor (or  stunt) .ratings or measurements    '

 ta cot    tf J' T ?ht'  dlafluster' «  l«»9th)  in  treated  areas should

 SveS*^6! ? Plants  in check  plots in  which commercially acceptable
 levels of pest control  are maintained.   Vigor ratings should be


 I?ir2Dort± ?ff °f mmXimm •ta»tl»9-   If -Bunting is observed,
 it is important that subseguent evaluations b« made to  document the
 oagree- of recovery.       .       -    .                •
 „„„ Uii) Pi"ttn! ^P^.'  A ran?e of P^ting depths' within the
 range recommended for the crop should be included ii preliminary


         "
                           Preemergence (to crop) applications.  Data

          from these 'trials should reflect any effecls of varying

 planting depths on .the' incidence of crop injury that might be


            r^T CaaM«i*1 »" condition,!-  L subset trials,

           T?  T    ^r^f1"*^ St '•«— ««« ***** settings should
           if  in preliminarv studies the  planting depth 1.' found to

       !1      ^*iafal«'  ="p emergence data should be taken 'from all





                          effect of pesticides on lodging of target

                                                            Arcane
                 ippr°Ximate  d«^«* °f  "9l« of lodging in treated
      should  be. compared  to  that in weed-free check plots.
(v)
           ghvtotoxieity.   Evaluations  of visible, symptoms of pesti-

=ide mw (such as discoloration,  ^formations,  Ssiccation

defoliation, or death) to crop plants should be  at  least visually

assessed and reported.  These symptoms  should be compared to results

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                         36
 in check plants untreated with the a«st±=H A. <„
 a^aas: sisS



£•=2 ,—                    -         ~
report^3 Hef? L*8 •'P^ici— onyi-ld. should be
        b8e<^eat Planting.  The effects of «.«*-)<-<<»


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                                 37

     (ii)  Rotational crops (including cover crops).  If detrimental
effects are observed, results of studies evaluating severity and-
duration of effects on the injured rotational crops should be sub-
mitted.  To determine the duration of phytotoxic effects,-susceptible
rotational crops should be planted at varying tine intervals after
pesticide application.  Such studies nay be combined, with field
studies designed to evaluate soil residues.  [See § 165-2 of Sub-
division N.]

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                                  38
Series 122:  TISR 1 OP NONTAHGET ASEA TESTING




                                                of
       Dieptyledon«m.i •   six
                                      -P.C1..
                                     ,# =,,.
                                                  3r=,»p,  should

                         •
                             that .,u.

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                                   39

  target area is significantly less than the maximum label
                                                           rate  .
                      °f ««l«l-t** the dose level « the' seed
                   '-  -       °  artW ia^edient P« «« should be
  considered to be equal to 3 ppmw in the solution which is applied
  to seeds.  (Notet a 1  lb. ai/acre application to a 3, inch soTl    '
  depth would equal 7.5  ppnrw in the soil solution.)


           '*?*   °  plaa^-  At 1«*rt 3 replicates,  each with
      st    =                     each *ith  at least 10 seeds,  should
 be  tested  per  dose, letrel  for the  seed  germination study.   Larger
 populations and »ore xeplicates may be needed :to increase the
 statistical -significance  of  the test,

 .H~ T I5 il ^'  Th* seed- -?ermin«ti«/s*«'iling  emergence  studies
 should be  conducted under controlled conditions in  growth chambers
 or greenhouses.  The vegetative vigor  test  may  be performed  in  a
 growth ehanber, greenhouse,  or  in small field plots.  *

      <«>  P««ti«i.  Ci)  Seed  gemination, if  performed  usina  petri
 plates or seed germination paper, should be assessed  after 5  days.
 Seedling emergence should be observed weekly/ or more frequently,
 for at least two ^weeks after germination.

      Ui)  The  effect of vegetative vigor should be .observed  weekly,
      -fe-;                      '     anora*  symptoms
      ,  the observations should be continued until the plant dies
 or fully recovers.     ,

      C7>   Protocols.  -Hie protocols for these tests outlining the
 acceptable environmental conditions ,  procedures , and some pertinent
 references. ..are found in § 122-30 (a) through (c).       '

    _  (e! I   Reporting,  m addition to  the information required in   -
 I  120-4 (b), the test report  should include the following iziforma-
 tion.             -                                   •


      (1)   The  number of seeds  tested  and the number germinated or
 emerged per dosage  level for each replicate;
ment  t**     *                          ***  ^he growth and develoo
ment of the seeds and emergent plants,  indicating any abnormalize.
and expressions of phyto toxic ity; and                  '

     (3)  Tabulation of the results indicating  the percentage
          Sl f°r MCh Sp*Clla as conf«red to  untreated control

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   also
                                    40

                            and height or other growth parameters  may
   . .'   {d>  Tieg orogrwioB.   (1)   2f  the results  of  the  seed
  germination/seedling  emergence  test(s) have  indicated an' adverse
  effect greater than 25 percent: on  one or more plant  species ,. then
  seed germination or seedling  emergence tests at the • Tier 2  level 'are
  reqmred, { see }- 123-1}.,',,   • •.        '.'      • •   ':''    .  V
  ,. i. A     lf-thm «salts of the vegetative vigor test(s) have  indi
  cated an adverse effect greater than 25 percent on one or more
  plant species,: then vegetative vigor tests at the Tier 2 level are
  required (see § 123-1).                            '

  _  _   (3)  If .less .than 'a 25 percent detrimental effect or response
  is noted for .either seed -germination/seedling' emergence or vegeta-
  tive vigor tests,  no additional testing of the respective tests
  at higher 'tiers is ordinarily required.  The Agency,  after review
  of tne data, may require certain additional tests to  determine a
  more definite nondiscernible effect level.
  $  122""2   Srovth and reproduction..ef aquatic 'giants (Tier 1) .


   _   U>  ' ?***  required..   (1)   Data on  the toxic .effects of a pesti
  c^»  on growth  and. reproduction of aquatic plants  are required by
  40 CFR Part  15:8 •aa_«-fiOfcbZrcM« . basis to .support the registration
  of each end-use product intended for outdoor pesticide application,
  and. each  manufacturing-use product which legally could be used to'
  make  such end-use products.   I See  §  120-1 (e).]

       (25  Studies of this section  need  not be conducted for pesti-
-  cides applied by systems where  the  chemicals  are not  readily
 released into 'the environment.   Examples of  these  systems are-
 tree injection, subsurface soil' applications, recapture systems,
 and wick applications..     '                        *''*'•

      (3)   Portions of this Tier  1 test may be combined with the
 respective parts of the Tier 2 test  {§  123-2) and  performed as  one
                    •
      (4)   See § 120-1 (e) concerning substitution of testing and
 data  submission requirements. '                      .     -   .  -

      (b)   7est standards. • In addition to the general test standa-ds
 set forth  in §,120-3, the following standards for the studies of  the
 growth  and reproduction , of aquatic plants apply:        •   '

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                                    41
         ILifT '!     ^-'   The tecimical 5r^« of the
           f   * .*•«*•   Where « technical grade does not exis
  the manufacturing-use product: or an end-use product with the h±«h.,t-
  percentage of  the  active ingredient shall be usedf *^h *h* hi?hest
        (2)  Species.   (j)" 'Selenastrum capt-i^r..,^ /.,' «T..^

           a) should 'be tested regardless of  the  intended outdoor Le



        (11)  If the intended use pattern' is -for. outdoor  aquatic  pest
            ...Lemna gibba .(-duckweed) ;    .....
             Skeletonena eostatum f marina '

             A freshwater diatom (unspecified species),, and
             Anabaena fles-aonae (blue-green alga).
  t6^J35,  ^ff1^8^011 levelS-  a« ^
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                                   42
Protocols. •' The  protocols for these tests

              C0nditi°-
              found in §



              addition to
            r                            Procedures aso
            references are found in §  122-3Q(d) through (h>?
                                                               by
  ml  J!L MS£* - Growth should b* expressed as the cell count per
  ml,  biomass  p«r volume, or degree of growth a« determined by         '
  spectrophotometric- means; and        '


       (3)  Tabulation of the results indicating the  percentage effect
  level versus tine as compared to the control.       Percentage effect



       (d}  ?ier Progression.  (1)   If a  detrimental'  effect 'or
 controls, Besting at Tier 2_is required.  See |  123-2.


      (2>  If less than a SO  percent  detrimental  effect or
 is noted  no additional testing at higher .rates         '
    t                            '               ra
 tests to determine a more  definite nondiscernible effect





 ^  r22~30   Aceeptalale methods, and reference.
                                =            r.-rr ss.
                             Chemical-   0*  — S- -7  be 'surface-
                                                 seeds
onti                         tCSt te»P*rature  should approximate the
optimum temperature for the  species  and  variety used.
                                  after^five days or more frequently.
     germination is reported  as  th* number of germinated seedT '

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                                   43
                        references.
            B°*awit*'  M'   1966*   * rapid. bioassay for PEBC and its
                 TOlatil4»«*°*  «* *i*orption studies .   jellies!
                         '                           The  use^of  three
                bi°*»say, on  forty-two herbicides.   weed Sas.  ii
       (iii) TrualOTa, B.,  (ed).   197?.  Research Methods  in
    mir      emerqenc<-  (1^  Protocol.   (ij  seeds may be'
        ft   fTS Antacid-wash^ .sand or -a  standardi^d soil.
    least 10 seeds per pot should be used.  .The  seeds -may be surfae
                                                      '
 treated with a known quantity of ~th* chemical.  The test
 should approximate those optimal condition, for thl sp^c
 ^arxeties considered.  The seeds should b« incubated for at- least
 Udays.  The seed, are observed after 10 and 14 day," and
 ««rgenc. is recorded as the number of emerged seedling,.
 Of  ^*   ^*;tMt M2T b« extended by 14 days to assess the effect
 of  •"^  «PPH«d-p««ticid«« on vegeta-tive vigor.    '    '

      ^2'   Reeomaended reference.                              •    '
Southl~r«rr!'cB;'  {€d)*   19?7'   Research Method, in weed Science.
Southern Weed  Science .Society.  Auburn Printing Inc., Auburn,  AL  221


, ,< (C)  H«g«tati*e vigor -foliar  gBMy.   (1)   protocol.   Ci)   ^
roiiar spray,can b*.applied, by .any acceptable method using  labora-"
°=ry-, greenhouse-, or-field-grown plant,. .  The  plant should be  1 *n
4 weeks post-eaarg^t in order to gain young folfag^?  ?Ss 2
^^ .M method, of, foliar applications may be found irfthe

         »xieT* (^r^iT^*1 effeCtS  are t0 be  reP°"ed  as severity
             snt /
                                                     of
   tne  seedling emergence study.        '

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                                   44

       (2}  Mconmended reference.  Truelove, B., (ed)'.  1977
       r=h Methods in weed Science.  Southern Weed Science Soci
  Auburn Printing Inc.,. Auburn, AL  221 pp.                  «=-

       (d)  Lemna qibba;  Growth conditions.  (1)   Species and
        |ibba G3.  Source, Dr. Charles Cleland,  Smithsonian Radia-
       Biology ;Labosatory,, Rocterill,,. MD ^20852  (limited supplier)

       (2)  Protocol.   The following are acceptable conditions for the
  growth and maintenance  of Lenma ' gibba G3.-'*

    ''  (i>    Environmental conditions.

             Light Intensity s   5  klox (approx.  100  uE aT2s-1}
             Light Quality;  warm  white  fluorescent
             Photoperiod?  continuous  light     ••   "
             Theraoperiodj   continuous  25 + 2"C

       Eii)  ^: '-'•sre conditions.  .          '

            Liquid culture    •   •
            Nutrieatss  M type -Hoagland's  medium without EDTA or
                        sucrose  (Hillman,  1961 a & b)
            pH S.O * 0.1 after autoc laving

      (114.) Procedures.  The vessel size- to-mediun quantity ratio
 should be S to 2.  Maintain the Leana stock under axenic conditions
 !!«!.*?*** %f7 ^ '^rfortt«1'un<5« aon-axenic conditions as long as hbn-
 organic media are used.   Sucrose £10 g/1)  and EDTA  (9 mg/1) may  be::t;
 added if flowering is desired.

                       references.
      (i)    Davis,  J.A.  1981.   Comparison of static-replacement and
 flow-through bioassays  using  duckweed, Lemna gibba-G3.  o.S.
 Snvironmental Protection Agency.   Washington DC  (SPA 560/6-81-003).

      (ii)   Hillaaa,  W.S.  I961a.   Experimental control of ' flowering
 in
          III.   A relationship between medium composition and the
opposite  photoperiodic  responses of L. oerpusyilla . 6746 and L.
gibba G3.   Amer.  J.-Bot.  4a;4l3-4i9.     .        •           • —

     (iii)  Hillaaa, W.S.   1961b,  The Lemnaeeae, or duckweeds. •
Bot. Rev. 27:221-287.       '.          - --
                       capricornutua:  Growth  conditions.   (1)  Soec' es
Selenastrua eapricornutua  Printz.   Source: EPA Corvailis Laboratory
Corvallis, OR  97330                   .   •                         y'


     <2\  Protocol .  The following  are acceptable  culture conditions
-or the growth and maintenance of Selenaatrua  caoricomuttia.

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                                   45

       (i)   Environmental  conditions..          .

            Light Intensity:  4 klux (approx. SO ut m-2t-tj
            Light Quality:  cool white -fluorescent
            Photoperiodj continuous light
            Thermoperiod: continuous 24 + 2"C

            Culture conditions.
                   culture
            Nutrients:  U.S. EPA (1978) medium (EDTA shall not be
                        used in the experimentation medium.-)
            pH 7.5    ___

      O>  Recommended references.

      U)   Environmental Protection Agency,  National Sntrophica-
 tion Research Program.   1971.   Mfal Assay Procedure:  Bottle Test
      BT*  Nit±0nal  •"'i'oa^ntal  Research Center,  Corvallis, OR
      (±1)  Millar,  wsfc,  j.c.  Gr««n
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                                   46

        £3)  Recommended references. .

      .U5   U.S. Environmental Protection Agency.  1973.  Bioassav
  procedures for the ocean disposal permit program,  g.s. SPA Labora-
  tory, Gulf Breeze, FL  32561 ( SPA-60 0/9-78-010 ).

        (ii)  Walsh, G.E., and S.V. Alexander.  1980.  A marine algal
  bioassay methods Results with pesticides and industrial wastes.
  Water, Mr,  Soil Pollut. 13:45-55. .

       (9)  A Fresnvater Diatom;  Growth conditions.  (1)  Soeci-s.  (To
  oe selected.}      . .         '       •                      "

       (2)  Protocol.   The following are acceptable culture conditions
  for the growth and maintenance of Navicula seminulum or other selected
  .freshwater diatom.  •  '        '                ~" - "

       (i>  Environmental conditions .

            Light intensity: 4.3  klux  (approx.  85 uE  m-2s-1)
            Light quality:  cool white  fluorescent
            Photoperiods   continuous light
            Thermoperiod :   continuous  24  +  2*C. ' '    '

            Culture conditions.-                   • '
                   caltar*
            Ntitrientss  U.S. EPA ,{ 1971 ) medium
            pH 7.5       • '             '                '

      (3-)  Recommended reference.                              '  •

      Environmental Protection Agency, -national Eu^rophication
 Research Program.  1971.  Algal Assay Procedure: Bottle Test
 (AA»:BT).  National Environmental Research Center, Corvallis,
 OR -97330      •                           '                 .

      (h^  Anaba«na flos-aquae; Growth conditions.  (1)  Species.-
 Anabaena flos- aquae (Lyngb.) DeBrebiason.  Source: EPA Corvallis
 Laboratory,  Corvallis, OR - 97330      '
           Protocol.  The following are .acceptable culture' conditions
.for the growth and maintenance of Anabaena flos-auuae.
                          conditions.
            Light intensity:   2 klux (approx. 40 uE m~2s~1)
            Light tjuality:   cool white fluorescent
            Photoperiod:   continuous lighr
            Theraoperiod:   continuous 24 * 2 "C

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                                 47,

     (ii)  Culture conditions.

           Liquid culture
           Nutrients:  U.S. EPA (1978) medium   (EDTA should not
                       be used in the experimentation medium.)
          , pH 7.5 (not to b« exceed 8.5)        "  .

     (3)-  Recomn.end.ed references.

     (i)   Carr, N.G., and B.A. Whitton, eds.   1973.  The Biology
of Bluegreen Algae.  University of California Press, Berkeley.
676 pp.   .   '         -    '              -     •

     (ii)  Environmental Protection Agency, National Eutrophication
Research Program.  1971.  Algal Assay Procedure: Bottle Test.
(AAPsBTJ. National Environmental Research Center, Corvallis,  OR
97330

     (iii) Miller, W.E., J.C. Greene, and T. Shiroyama. ^1978.  .The-
Selenastrum eapricornuturn Printz algal assay bottle test.  D.S.
Environmental Protection Agency, Corvallis, OR  97330 (EPA 600/9-78-
018).                             '    •       .-. .          -

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                                   48

  Series 123s  TIER 2 NONTARGST AREA TESTING




  * 123~1  Seed germination/seedling emergence -and veget
                                          -- ;
               r 2
                 required.  (1)  Additional data on the phytotoxic
  effects of a pesticide on seed germination/ seedling emergence or
  vegetative vigor, respectively/ are required by 40 CFR Part 158 on
  a case-by-case basis when a 25 percent^phytowxic effect S one
  or more .plant species is noted as -a result of the respective Tier
  1 tests.   These data are required. to " support the registration of
  each  end-use product intended 'for outdoor application'..

      .(2)   Portions  of this Tier 2 test may be combined with the'
  respective -parts  of the  Tier 1  test (§ 122-1) ;and performed as one
  &6S-t»        •              •'

       (3)   See  §  120-1 (e)  concerning substitution of  testing and data
  submission requirements.              •          .                 "«&«

       (*l   7es*  standards.  In addition to  the general  test standards
 set forth  in §  120-3,  the  test  standards  for  this  section  shall, be
 the same as those contained . in  the  Tier 1  studies  If  122-Kb)]  with
 the following modifications:                           -

      (]}  P°«*?*«-  Th« following dosages  should' be  tested:   (i)   At
-least 5 dosages should b«  tested?

      (ii)   2-he dosages should include  a subtoxic  «EC50) and a  non-
 toxic concentration;

      (iii)  The highest dosages should be less than the -1-fold dosage
 tested in  $ 122-1
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J
                                                  49

                is greater than  the  BC25  for one  or .more
                  123"2  Grovtn a**3 reproduction of aquatic plants
                 __  i,,aV0'*9"* Sh0llld ^ °f 9eoffietei= Progression of no more
                    ff  ?*-   '" ««»Pl«» the test concentration series may be-  0.1
                .2,  0.4,- 0.8,  and 1.6 kg/ha/15 cm (a 2-fold progression).
               r/,   (2)   PIant «o«gi".  At least those plant species of Tier  1

                    22"1211  %*iCh «hiiite
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                                   51

  Series 124*  TZ2R .3 NOtnSURGET AREA TESTING '       "         •

              T


  5 124~1  Terrestrial field tasting (tier 31.   •


  _u   (*}  • WMa g«T"*g«fl»  CD   Data on the phytotoxic effects of
      •»!-«• product on Mad germination, vegetative vigor, and
  reproduction potential under field use conditions ara required by
  ~S SJhP*!L15a °n ! c*»«-by-c*~  *•«*• to support the registration
  of each end-use product intended for outdoor -application,   the
  a«i«u=. racomesded application quantity or anticipatad environ-  -
  mantal  axposure ia to ba actual  to  or graatar than tha BC2S for ona
  or aora tarrastrial plant species  as found in  tha Timr 2 ta«ts
  (|  123—1) »


  „,, i2i  Th* idata r«Ittir«*=*s  °«  thi.  section need- not ba ful-
  filled  for pesticides  applied by systems  where tha chamicals are
  not readily released into  the ararironnent.   Exaaples  of these
  systems are:  tree  injection, subaurface  soil applications,  recap-
  ture systems, and wick applications.'   *

      (3)  See }  120-1 (e) conceraing substitution  of testing and  data
 requirement submission.                                    *
 set fo^h !!*! Sf*;'  fc Mliitloa to th. general test standard,
 sat forth in § l2§-3, tha test standards for this section shall be

 S! JT' S th!^,!?ntalJl*d ^ f 122-1(b> «« ^» subdivision, with
 tha followiaf modifications!
    „ (1)   ?Mt substance.  The test substance shall be the end-use
 product or a representative end-use product from the same major
 formulation category for that general use pattern.  Sxamples of
 major formulat^m categories aret wettable powders, emulsifiable
 concentrates,, and granular*.  (If the manufacturing-use product is

 Mjo^LlSiSr1 iat° *ad"U" V*0*"** <=°»pri*inrtw6 oTaS
      a typical end-use product for each category.)     **  °m
 -  „. C2)  *PP^.c«tion- levels,   fhe dosages tested should be the same
as those employed  in  the  tier  2 test [f  123- Kb) ( 1 )] .

_  t3)  SiSSiM*- (i>   Hapresentatives  of the following plant

UiiTbalowf * fe**t;6d'  *Ui'J*Ct *° th*  llmit*tion« of paragraph


      (A>  Oicotvledonae (dicots) ,  representatives of three families?
     (B)  Monocotvledonae  fmot,^.^.^ r representatives of three
families}

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                                   52

       (C)
  two famili                                        '        naves of.


       ^D)  33T°phyt« Cesses) or HepatODhyta_ (liverworts), one
  sentative (for wetland use., pattern* only )~and

       (2)  gymnospermae (conifers), one representative.  '

       Cii)  Plant species . used for testing Tiers 1 and 2 can be used
  to satisfy the aonocet or -dicot. test plant requirements of this'
          "    "      -                  •            "••
  to           t                             not likely to be exposed
  to the pesticide under normal conditions' of use,  'testing of such
  groups is not required.  Justification for elimination of a test
  species or group should be included in the test report.
  ,.,  ,            u     -       sP«ci«  ^y  b«  required if the general
  selectivity  of  the  pesticide cannot  be readily  identified.
     J4)  ^st  conditions, "giants  are to be grown under field-use
 conditions similar to  those of the  natural habitat of  the plants  in
 use.
      ^5}  Duration.  The test duration should be of  sufficient  length
 -to assess multiple applications directed by the label.   Obser^tioS
 should continue^ for at least two weeks after 'the last application  and
 for * maximum of four week* to note any recovery or  death.  '
      (6)  Sea**?" °f application.  The test substance is- to be applied
 over a period of time or season according to. the proposed label
 instructions .                .        .          -  . •     .

      (7^.  T««* locations.  The pesticide. -should "be tested in .those
 geographic locations where it is expected, to be used, as based on
 proposed label use sites.  Where important species diversity and
 physiographic differences occur within a region of intended' applica-
 tion,  regional testing may.be inadequate, and testing at a more
 specific region or biome level may be required.  United States
 regional areas of  potential testing' include:

     Northeastern  temperate deciduous;    '-'               '
     Southeastern  temperate • deciduous;
     Northern  grassland. {prairie );
   -  Southern  grassland ( prairie},- '
     Northwestern  (and Alaskan)  conifer  forest and high desert;
     Southwestern  chaparral Mediterranean and  low desert;  and
     Hawaiian  and  Caribbean tropical  regions.

     Cc).  Reporting,.   In addition  to  .the  information required in
§§ 120-4 and 122-1(c}  of this subdivision, the  test  report  should

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                                   53
  include  the  test  condition* employed (including the soil and
  environmental  conditions)  and the determination of 'the 50 percent
  detrimental  effect  level.                   •    '          F«^c«Jt
 5 124~2  ^^uatic field testing (Tier 3).


      (a)  ***** required.  {1)  Data on the phytotoxie «ff«cts of the
 product on growth and reproduction of an expanded number of aquatic
 plants are required by 40 CFR Part 158 on 'a case-by-case basis to
 support the, registration of each end-use product intended for outdoor
 pesticide application, whens

      (i)   The anticipated environmental exposure is greater than the
 SCSO for any one aquatic plant species tested in Tier 2 tests {§ 123-
 2);  and                                  '-....

      Cii)   The pesticide is expected to be applied' to a fresh water,
 estuarine,  or. marine aquatic system by -either direct application ox-
 direct  discharge of treated water (except swisHing pools) , or the
 pesticide  is to be used within a forest system.  [See f 160-3(c),
 (d), and (e)  of Subdivision R for descript±ojss-of these aquatic uses.l
 Pesticides- with only terrestrial uses need not be tested.

      (2)   See | 120- 1(e)  concerning substitution of testing and data
 requirements  submission.

      (b>  ***** «*«pq*rds.   In addition to the general test standards
 set  forth in $  120-3 of  this subdivision, the test standards for this
 section shall be  the same  as those  in f  122-2(b), with the following
 modifications!

      C1)  T**t  «ofeyt*nce.  The  test substance shall -be the end-use
 product or a ' representative . end-use . product from the same  major
 formulation category for that -general  use pattern.  Examples of
 major formulation categories ,arei wettable  powders,  emulsifiable '
 concentrates, aad graamlars*  (If the  manufacturing-use  product is
 usually formulated into end-use products  comprising two  or more major
 formulation categories, a  separate  study  must be performed " with a
 typical end-use product for  each category.)              .  -
          Application levels.  The dosages tested  should be  the sane
as those specified in the Tier -2 aquatic test  standards  [5  123-
Kb) nn.                                               -  5

     (3)  SP«cie_s.  (i)  Aquatic plant representatives of the
following plant groups are to be tested-?

          Pieotyledonae  (dicots) , one representative?

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                                    54
   families, -n°COtVled0na* <*«»«*•>' representatives of ' three
   of
  Division,                 *  -°E>,  a representative of each
            Bryophyta  (aosses) or
       (ii)  Plant species used for testinc- Tiers
             the
       (iii) Additional plant species may be recniired if
  selectivity of the pesticide cannot
                              °r
                                          by naturU or
             ..y. b. n.c.,.ary  to  .i»ul»t. the  reduced l-
                                                          c«txoll.d
                                                                  ••
      (Y).  .The. field studies should be
                                        conducted using:
southds                 Ci ^  Methods  in weed science, 2nd Ed.
Southern rfeed Science Socx«y,  Auburn  Printing inc.,  Auburn,  AL;  or

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                                 55

      (B)  & protocol with prior approval of the Agency.

                     'Th*> teat duration should be of  sufficient
length to assess multiple applications directed by the  label*
Observations should continue for at least two weeks after the  last
application and for a maximum of four weeks to note any recovery
or death. '

     (6)  Season of application .  The test substance  is to be  ap-
plied over the period of time or season according to 'the proposed
label instructions.

     {7}  Test locations*  The pesticide should be tested in those
geographic locations where it is expected to be used, as based on
proposed label use sites*  Where important species diversity and
physiographic differences occur within a region of intended appli-
cation, regional testing may be inadequate, and testing at a more
specific, region or biooe level say be required.  United States
regional areas- of potential testing include:

     Northeastern temperate deciduous;
     Southeastern temperate deciduous i
     Northern grassland { prairie) j
     Southern grassland (prairie);
     Northwestern (and Alaskan) conifer forest and high, desert;
     Southwestern chaparral Mediterranean and low desert} and
     Hawaiian and Caribbean tropical regions.

     (c)  Reporting.  In addition to the information  required  by
§f 120-4 and 122-2(c) of this subdivision, the test report should
include the test conditions ( including soil , water, and environ-
mental conditions) and the determination of -the 50 percent detri-
mental .effect level.'

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        P&CS
       Pesticide Assessment Guideline, Subdivision J
       Hazard ^valuations  Nontarget Plants .  ,
       Robe
           Hoist and ThomasC.  Sllv*naer_
Office of Pesticide  Programs
"J.S. 'Environmental Protection Agency.
Washington, 3«C«   20460
                                                               4. »»oort C»t«
                                                                    October 27,  1 982
                                                                     10.
                                                                     JI. tswtnMsttC ar c/»m(O)

                                                                     (G
12.
                     tna
      Office  of ?*sz;icide Programs
      "J.3.  SavirsnauBRtai ?roracrion  Agency
                                                                  'ro» a* atoart 4 »•«•«' .Co«»r»a

                                                                   Guideline
         iiincssn,  O.C.  204SO
                                                                     'U.
                ,
     3-uid«lines Project Manager  :  Robert K. Hitch
  Ao»t.-ict i
            Suisdivision J, a Federal Insecticide,  Fiangicide  and, Rodenticide Acr
       (FISTUV) .guideline, prescribes the phytotoxicity  sttody protocols that the
       U.S.  Environmental Protection Agency recommends  to  support the registration
       of manufacforing-oae- and formulated end-use products.   This subdivision
       establishes procedures for testing and data submission concerning the effects
       of pesticides on nontarget plants.  The studies  outlined by this subdivision
       will  not  be required for.every product but  only  for selected chemicals on a
       case-by-case basis.  A stannary of the rulemaking phytotoxicity test requirements
       can "be  found in 40 CPR -Part 158.  The results, of the  phytoxicity studies
       together  with environmental fate and efficacy are used to assess the potential'
       hazard of pesticides on'terrestrial and aquatic  nontarget plants.

            Subdivision J constitutes one volume of  a twelve-part FIFHA guideline
       series published by the National Technical  Information Service.

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