EP A BIOPESTICIDE S AN D POLLUTIO N PREVENTIO N DIVISIO N

COMPAN Y NOTIC E O FFILIN GFO RPESTICID E PETITION S PUBLISHE D I N TH E FEDERA LREGISTE R EP A Biopesticide s an dPollutio nPreventio n Divisio ncontact : [Kimberl y Nesci , 703

- 308-8059 ] SUBMISSION : E-mai l th e complete dtemplat e to : reilly.sheryl@epa.gov . TEMPLATE : [Andermat t

Biocontro l AG ] [Inser t petitio n number ] EP A ha s receive dapesticid e petitio n([inser t petitio n number] ) fro m [Andermat t

Biocontro l AG] , [Stahlermatte n 6 CH-614 6 Grossdietwi l Switzerland ] proposing , pursuan t

t o sectio n408(d ) o f

th e Federa l Food , Drug , an dCosmeti c Ac t (FFDCA) . 2 1 U.S.C . 346a(d) , t o amen d4 0 CF Rpar t 180 . (Option s (pic k

one ) 2 . t o establis ha nexemptio nfro mth e requiremen t o f atoleranc e fo r

Autograph a californic amultipl e nucleopolyhedroviru s (AcMNPV ) (Option s (pic k

one ) 1 . microbia l pesticid e \Autograph acalifornic amultipl e nucleopolyhedroviru s (AcMNPV) ] • • ••••• • • Pursuan t t o sectio n408(cl)(2)(A)(i ) o f FFDCA , a s amended , [Andermat t ;...; . Biocontro l AG ] ha s submitte dth e followin g summar y o f information . daftf-M M argument s i nsuppor t

o f thei r pesticid e petition . Thi s summar y wa s prepare dRy«[SciR«g , • Inc. ] an dEP A ha s no t

full y evaluate dth e merit s o f th e pesticid e petition . Tffi ? Summar y • • ma y hav e bee nedite db y EP A i f th e terminolog y use dwa s unclear , th e summar y *** • ' containe dextraneou s material , o r th e summar y unintentionall y mad e th e reMJe ? conclude , tha t th e finding s reflecte dEPA' s positio nan dno t th e positio no f

th e petitioftft f

. • • • I , [Andermat t

Biocontro l AG ] Petitio nSummar y • IA . Produc t Nam e an dPropose dUs e Practice s [A n end-us e materia l containin g Autograph acalifornic amultipl e nucleopolyhedroviru s (AcMNPV ) i s bein g submitte dt o EP A fo r registration . Loope x wil l b e applie dpre-harves t i n greenhouse s an dope n field s t o col e crops , gree n leaf

y vegetables , celery , beans , peas , peppers , cucumbers , tomatoes , peppers , melons , ornamentals , an dflower s t o protec t thes e crop s agains t

infestatio n cause d b y cabbag e loope r

larva e (Trichoplusi ani) . Applicatio n rate s ar e projecte dt o b e u p t o 5 flui dounces/acre. ] B . Produc t

Identity/Chemistr

y 1 . Identif y o f th e pesticid e an dcorrespondin g residues . Autograph acalifornic a MNP V stai n F V #1 1 Physical/Chemica l Propertie s Propert y Colo r Physica l Stat e

Odo r Densit y P H Viscosit y Resul t

Ligh t mediu mbrow n Liqui d

Undetectabl e 1 . 1 5 4 g/ m1(9.6 3 Ibs/gal ) 6.3 1 

### 12.64cStat20°C . 

### 5.8 6 cS t a t 40 ° C . 

### Referenc e Sylva r Technologies , In c Report , Loopex ; Autograph acalifornic a multipl e nuclepolyhedroviru s (AcMNPV ) F V # 1 1 , batc h# 6 Physica l Propertie s Sylva r Technologies , In c Report , Loopex ; Autograph acalifornic a multipl e nuclepolyhcdroviru s (AcMNPV ) F V #11 , batc h# 6 Physica l Propertie s Sylva r Technologies , In c Report , Loopex ; Autograph acalifornic a multipl e nuclepolyhedroviru s (AcMNPV ) F V # 1 1 , batc h# 6 Physica l Propertie s Stillmeado w Produc t

Chemistr y Report , Stud y No . 1 7287-1 3 1 . . . I . Stillmeado w Produc t

Chemistr y Report , Stud y No . 17287-1 3 *•••*• • Stillmeado w Pro dfi e t'&ntmistr y 9 Report , Stud y No . 3 72 & 7*1 3 " . . .



## ••:•• : : *. .

••••• • 2 . Magnitud e o f

residue s ti l th e tim e o f

harves t an dmetho duse dt o determin e l m K residue . [N o magnitud e o f residu e (MOR ) studie s hav e bee n conducte dwit h Autograph acalifornic aMNPVstrai n F V #1 1 base do n th e lo w toxicit y o f th e activ eingredien t (se e Toxicolog y Profile ) an datota l absenc e o f an y know n infrection s t o organism s othe r

tha n anaroo w rang e o f Lepidoptera n insects . OPPT S Guidelin e 885.2000 , sectio n f, Backgroun dfo r Residu e Analysi s o f

Microbia l Pest

Contro l Agent s (MCPA) , states : "Th e Agenc y evaluate s residu e dat afo r MCPA s use do n food , feed , o r ra w agricultura l commoditie s onl y i f toxi c o r othe r harmfu l propertie s wer e observe di n th e maximu mhazar dtoxicolog y test s (Tie r 1 ) prescribe di n toxicolog y tes t guideline s OPPT S 885.305 0 throug h885.365 0 o f thi s series . I f

Tie r 1 toxicolog y test s indicat e n o toxi c o r

othe r harmfu l properties , the n n o residu e dat a(wit hth e genera l exceptio n o f

amonitorin g method ) woul db e indicate dan dthu s arecommendatio n fo r a n exemptio n fro mth e requirement s o f atoleranc e ca n b e made." ] 3 . A statemen t

o f wh y an analytica l metho do f detectin g an dmeasurin g th e level s o f th e pesticid e residu e ar e no t needed . [Andermat t

Biocontro l A Gi s submittin g a petitio n t o establis ha n exemptio n fro mth e requiremen t o f atoleranc e and , therefore , a n analytica l metho di s no t required. ) C . Mammalia n Toxicologica l Profil e [Th e ful l report s tha t

suppor t thi s petitio n an dwhic har e summarize di n thi s sectio n wer e previousl y submitte dto , hav e bee n reviewe dan daccepte db y th e Agency , an dcite dherei n i n suppor t o f anothe r simila r baculoviru s product , Gusano , EP A REg . N o 70051-43 . Andermat t

Biocontro l A Grelie do n toxicity/pathogenicif y dat acite dfro mvariou s source s o f

publi c literature . Base do n th e dat asubmitte dan dcited , i t i s establishe d tha t Autograp hacalifornic aMNP V strai n F V #1 1 i s neithe r toxic , no r

pathogeni c no r pathogeni c t o mammals , birds , fish , aquati c invertebrates , plants , an dnearl y al l non-Lepidoptera n insects .Acut e Ora l Toxicity/Pathogenicity :

Study 1 (MRID 74175) : Twenty

- mal e an d2 0 femal e Sprague-Da w le y rat s wer e dose dpe r o s wit hAutograph acalifornic aMNP V at arat e o f

4.3 5 x 10 ' occlusio n bodie s pe r

rat . Tes t animal s exhibite dbod y temperatures , mea nbod y weights , an d foo dconsumptio nsimila r t o tha t o f th e controls . Fou r

animal s o f

eac hse x wer e sacrifice do n th e followin g days : 0 , 3 , 7,14 , an d21 . Hematologica l an dbloo d chemistr y result s appeare dt o b e withi nnorma l limits . Histopathologica l examinatio nreveale dn o microorganis mrelate dlesions . Ther e wer e n o death s an d al l animal s appeare dnorma l throughou t th e entir e study . I t

wa s conclude dtha t

A californic aMNP V wa s no t injuriou s t o rats . Study 2 (MRI

D 43912201) : A s cite dpe r th e Nationa l Pesticid e Informatio n Retrieva l Syste m Acut e Derma l Toxicity : Study 1(MRI

D 72638) : Fiv e mal e an dfiv e femal e albin o guine apig s wer e treate d wit hasingl e derma l applicatio no f 0. 4 mLo f

suspende docclusio nbodie s (4.3 5 x 10 7 occlusio nbodie s pe r

rat. ) o n thei r

shave dbacks . Dose s o f

0. 1 m Lwer e applie dt o 2 abrade darea s an d2 area s o f intac t ski no n eac hanimal . Ther e wer e n o death s an d al l A californic aMNPV-treate danimal s appeare dnorma l throughou t th e entir e study . I t wa s conclude dtha t

A californic awa s no t

injuriou s t o rat s an ddi dno t produc e derma l toxicity . Study 2(MRID 48624712) : A derma l toxicit y stud y wa s conducte dwit hth e tes t materia l Baculoviru s heliothi s NPV . Rhesu s monkey s wer e expose dt o th e organism . Al l monkey s excep t

fo r on e survive dth e stud y an dshowe dnorma l w eigh t gains . On e femal e monke y i n th e subcutaneou s injectio ngrou p die do f unknow ncause s a t da y 2 4 o f th e study . On e subcutaneou s injecte dmal e ha drapi dsedimentatio nrate , elevate dlecocyte s an dreduce dlymphocyte s o n da y 18 1 whic hreturne dt o norma l o nda y 197 . N o tes t

substance-relate deffect s wer e observed . Acut e Pulmonar y Toxicity/Pathogenicity" : Stud y 1 (MRI D 48624712) : A n acut e pulmonar y toxicity/pathogenicit y stud y was * conducte dwit hth e tes t

materia l Baculoviru s heliothi s NPV . Rhesu s monkgv s weft*** * expose dt o th e organis ma t 1 mg/k g b w (1. 2 x 10 8 PIB/k g bw) , I x weekj y fi>i*2 6 • weeks . A H monkey s excep t fo r on e survive dth e stud y an dshowe dnorjnaj}\eigh{* . * • gains . On e femal e monke y i n th e inhalatio ntreatmen t rout e ha dlo w plalek i a t dfav"* . 18 1 whic hreturne dt o normal . N o tes t substance-relate deffect s wer e ojljs,e,r^'ed . *• • 

## : : 

## • •••• Acut e Inhalatio nToxicity : I • • Study 1 (MRI

D 74l76):Te n mal e an dte n femal e Sprague-Dawle y rat s wer e exposed**• • t o asingl e dos e o f

6 x 10 1 1 polyhedra l occlusio nbodie s o f AcMNP V fo r

on e hour .Th e nomina l chambe r concentratio n afte r

on e hou r wa s 11. 1 mg/L . Al l animal s wer e observe dfo r

mortality' , sign s o f

toxicity , an dabnorma l behavio r throughou t

th e one-hou r exposur e perio dan ddail y fo r 1 4 day s followin g exposure . Bod y weights , foo dconsumption , an dorga n weight s wer e als o observed . Inhalatio n exposur e o f rat s t o AcMNP V produce s n o injuriou s results . Ey e Irritation : Study 1 (MRID 72639) : A singl e applicatio n o f 0. 1 m l o f eac hth e tes t

materia l wa s administere dint o th e conjunctiva ! sa c o f th e lef t ey e o f eac ho f fiv e mal e an dfiv e femal e albin o (Ne w Zeaan dWhite ) rabbit s weighin g fro m2. 1 t o 3. 3 kg . Th e righ t ey e o f eac hrabbi t

serve da s acontro l receivin g asingl e application , 0. 1 ml . o f a 0.8 % salin e solutio n containin g 2 % lactose . Initia l fluorescei n examinatio n confirme dth e absenc e o f irritatio n an dcornea l damag e i n al l eyes . Observation s fo r i njur y an dirritatio n wer e recorde da t 24 , 4 8 , an d7 2 hour s an dseve n an d1 4 days . N o ey e irritatio n wa s observed ; th e ey e irritiato n scor e wa s zero . Derma l Irritation : Study 1 (MRI

D 74177) : Derma l irritatio n wa s assesse db y applyin g 0. 5 mL(2.1 7 x 10 7 polyhedra l occlusio n bodies ) ofAutograph acalifornic aMNP V t o intac t an d abraide dski n o f

si x Ne w Zealan dWhit e Rabbits . Assessmen t followe dth e Draiz e system . N o derma l irritatio n wa s observe dand , therefore , th e primar y irritatio n score wa s zero . Study 2 (MRI

D 48624717) : A primar y derma l irritatio n stud y wa s conducte dusin g th e tes t substanc e Granupo mS C (CpGV ) containin g 2. 2 x 10 1 granules/L . Th e tes t substanc e wa s applie dundilute dder mall y i nasingl e dos e o f 0. 5 mL/anima l ont o th e shave ddorso-Iumbe r

regio n o f

3 rabbits . Th e applicatio n sit e wa s covere dwit ha patc han dsecure db y a n occlusiv e bandage . Afte r remova l o f

th e bandag e a t

th e en d o f th e exposur e perio d( 4 h) , th e residua l tes t substanc e wa s gentl y remove dwit h water . Observation s fo r

mortality , behavior , clinica l signs , bod y weight , an dski n reaction s wer e made . Durin g th e 10-da

y observatio n period , n o mortalitie s an dn o systemi c toxicit y J...J . symptom s wer e noted . Behavio r wa s unaffected . Th e bod y weigh t gai n wa s i nthf

e norma l range . N o sign s o f ski n irritatio n wer e foun dafte r th e 4 h-exposurejf>eri<Jd? • • •

• • • * Granupo mdoe s no t

produc e an y derma l irritatio n i n th e rabbit . Th e facj^o/anyj* . * . sign s o f irritatio n indicate s Granupo mi s Toxicit y category ' I V fo r derma l iiiritatHMi. '

* * Hypersensitivir y Incidents : N o report s o f hypersensitivit y hav e bee n ass&ci&e dwith * Autograph acalifornic aMNP V strai n F V #11 . '....'. .Cel l Culture :

Study 1 (MRID 1431081 : Thirty-fiv e nontarge t hos t cel l lines , 2 3 o f

huma n an d1 2 o f nonhuma n vertebrat e origin , wer e expose dt o Autograph acalifornic amultipl e nucleopolyhedrosi s viru s preparation s derive dfro mfou r differen t sources : polyhedra , hemolymph,cel l cultur e medium , an dculture dcells . Th e viru s an dcell s wer e incubate dtogethe r a t tw o differen t

temperatures , 2 8 o r

37°C , fo r

fou r differen t length s o f

time , 16 , 40 , 64 , o r

16 8 h , an dth e cell s wer e assaye dfo r th e presenc e o f

viru s b y aperoxidase-antiperoxidas e detectio n method . Th e estimate d sensitivit y o f

th e assay a s routinel y conducte dwa s 0.9 8 n g o f alkali-liberate dvira l protei n an d1.95 n g o f

budde dvira l protei n pe r mm2 . N o evidenc e offran k replicatio n wa s obtaine di n an y o f th e 3 5 cel l line s tested , althoug hviru s uptak e appeare dt o b e quit e common . Viru s uptak e wa s confirme di n som e case s b y electro n microscopy . Th e degre e o f viru s uptak e appeare dt o b e dependen t o n cel l type , tim e an dtemperatur e o f incubation , an dvira l phenotype . Viru s purifie dfro m polyhedr awa s generall y take n u p mor e readil y tha n wer e th e othe r form s tested . Study 2 (MRI

D 143109) : Autograph acalifornic amultipl e nucleopolyhedrosi s viru s (AcMNPV ) wa s examine dt o determin e whethe r th e viru s o r

it s DN A coul dreplicat e and/o r persits t i n culture s o f

mammalia n cells . Huma n HeL acell s o r primar y huma nembryoni c kidne y cells , simia n CV 1 cells , hamste r BHK2 1 (B3 ) cell s o r Muntiacu s muntja k

cell s growin g i n monolaye r culture s wer e use di n thes e studies . Cell s wer e inoculate dwit hAcMNP V a t multiplicitie s rangin g fro m0. 1 t o 10 0 PFU/cell . Subsequently , th e inoculate dcell s wer e investigate dfo r viru s productio n an dfo r th e replicatio n an dth e persistenc e o f vira l DNA . Extract s o f

inoculate dcell s wer e als o screene dfo r

th e occurrenc e o f

AcMNPV-specifi c RNA . AcMNP V doe s no t multipl y i n an y o f

th e cel l line s studied . Vira l DN A replicatio n o r transcriptio n coul dno t b e detecte db y blottin g an dnuclei c aci dhybridizatio n experiment s usin g nick-translated , clone dvira l probes . Furthermore , ther e wa s no t evidenc e fo r

th e persistenc e o f

vira l DN A o r

o f fragment s o £vira l DN A i n mas s culture s o f mammalia n cells . Study 3 (MRID 147020) : Mammalia n cel l line s fro mChines e hamster , India n muntja c an dmous e wer e inoculate dwit hinfectiou s supernatan t o f Autograph a californic a(Speyer ) multipl e nucleopolyhedrosi s viru s (AcMNPV ) replicate di n ;...; . Mamestr abrassica

e (L. ) cel l culture s (IZD-Mb-0503) . Ther e wa s n o advers e effec t o n cel l proliferation , no r

wa s acytopathogeni c effec t (CPE ) induce di n suc hculture^ ? • • • • Cytogeneti c dat aindicat e tha t uptak e o f

AcMNP V int o th e cytoplas mJ> f J I • mammalia n cells , a s show n b y a n electro n microscopi c study , induced'neithe r ;• . * . numerica l o r

structura l chromosom e aberration s no r siste r chromatic ! etch&nge " (SCE ) events . • ••• • • • D . Aggregat

e Exposure §•••• • 1 . Dietar y exposure . [Th e potentia l fo r non-occupational , dietary , an dnon- « dietar y exposur e t o Autograph acalifornic aMNP V b y th e genera l population , includin g infant s an dchildren , i s likel y throug hinges t io no f treate dcrops. ]1 . i . Food . [Dietar y exposur e t o Autograph acali f arnic aMNP V shoul dno t b e o f concer n du e t o th e lo w toxicit y show n acros s th e entir e toxicologica l profil e (se e above) . I f Autograph acalifornic aMNP V i s ingested , non e o f th e studie s hav e indicate dan y pathogenicit y o f th e organis mi n mammals . I n addition , AcMNP V doe s no t replicat e i nmammalia n cells. ] ii . Drinkin g water . [Autograph acalifornic aMNP V doe s no t pos e aris k

t o drinkin g water . Anticipate dagricultura l practic e prohibit s direc t applicatio n o f

th e end-us e produc t

directl y t o water . Autograph acalifornic aMNP V i s no t likel y t o b e presen t i n aquati c environment s sinc e i t i s no t know n t o inhabi t

aquati c environments. ] 2 . Non-dietary exposure . [Non-dietar y exposur e t o Autograph acalifornic a multipl e nucleopolyhedroviru s wil l b e muc hmor e likel y throug hhandlin g b y formulator s an dapplicators , bu t i n ligh t o f th e subject s toxicologica l profile , shoul d b e o f littl e concern. ] E . Cumulativ e Effect s [Sectio n 408(b)(2 ) o f

th e FFDC A require s that , whe n considerin g whethe r t o establis ha n exemptio n fro matolerance , th e Agenc y conside r availabl e informatio n concernin g th e cumulativ e effect s o f th e particula r

ingredient' s residue s an dothe r substance s tha t

hav e acommo n mechanis mo f toxicity . Thes e consideration s includ e th e possibl e cumulativ e effect s o f suc hresidue s o ninfant s an dchildren . Autograph acalifornic aMNP V i s effectiv e b y dissolvin g int o th e larva l midgu t an d releasin g th e occlusio n derive dvira l particle s (ODVs ) tha t initiall y infec t

midgu t tissue s i n orde r

t o systematicall y infec t

an dreplicat e i n th e nucleu s o f othe r tissue s withi nth e larvae . Th e infecte dlarva e sto p eating , becom e sluggis han ddi e o f vira l infection . Followin g death , th e larva e decompose s an dth e progen y viru s i s release d i nth e surroundin g environmen t an dca n i n tur n infec t

neighborin g cabbag e loope r larva e (Trichoplusi ani)_followin g ingestion . Baculoviruse s co-evolve dwit han dhav e onl y bee n foun di n insects . Baculoviruse s no t infec t Hymenoptera n pollinator s an dparasitoid s o r insec t predator s suc has * prayin g manti s an dladybu g beetles . N o membe r o f th e Baculovirida e famil y ha £ bee n foun di n vertebrate s o r plants , an dAcMNP V doe s no t replicat e p i n}a}nmalian cells . Baculoviruse s d o no t affec t o r infec t fish , wildlife , humans , o r be'neficfa l ;• . • . organisms . Thi s passive mod e o f actio n doe s no t

hav e acommo n che micj j J-llase d* mechanis mo f

toxicit y a s ca nsometime s b e foun dwit htypica l organicjch^icals. l *.* , /'" . Safely Determinatio n ,•••• • i . U.S . population . [Autograph acalifornic aMNP V exhibit s lo w mammalia n *.,*. ! toxicit y a s demonstrate db y th e toxicologica l profile . Whe n Autograph acalifornic aMN.P V i s use do nra w agricultura l commoditie s i naccordanc e wit hgoo d

agricultura l practice , i t

meet s th e reasonabl e certaint y o f n o har mrequirement. ] 2 . Infant s an dchildren . [I t i s believe dtha t

th e potentia l exposur e o f infant s an dchildre n t o residue s ofAutograph acalifornic aMNP V throug hthei r us e t o protec t plant s an dfrui t

agains t cabbag e loope r

larva e i s n o greate r

tha n tha t o f th e genera l population . Availabl e informatio n doe s no t sugges t tha t infant s an dchildre n ar e mor e susceptibl e t o th e effect s ofAutograph acalifornic aMNPV ) tha n ar e adults. ] G . Effect s o n th e Immun e an dEndocrin e System s [Extensiv e literatur e searche s an drevie w ofAutograph acalifornic aMNP V toxicit y dat aindicat e ther e i s n o evidenc e tha t i t i s a n endocrin e disrupte r o r tha t i t ha s an y know n effec t

o nimmun e system s a s AcMNP V doe s no t replicat e i n mammalia ncells. ] /•/ . Existin g Tolerance s [Ther e ar e n o know n curren t agricultura l tolerance s o r toleranc e exemption s fo r Autograph acalifornic aMNPV . A previou s exemptio n fro mth e requiremen t o f a toleranc e fo r

polyhedra l occlusio n bodie s ofAutograph acalifornic aMNP V (4 0 CF R 180.1125 ) wa s revoke di n Apri l 200 4 base dsolel y o n non-paymen t o f maintenanc e fee s an dbecaus e ther e wer e n o activ e FIFR A produc t registration s applicabl e t o th e exemption. ] / . Internationa l Tolerance s [Ther e ar e n o know n internationa l tolerance s fo r Autograph acalifornic a MNP V althoug hexperimenta l an dcommercia l us e o f AcMNP V i s reporte dfro mCentral Americ a(1998). ]

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