## Buye rCropSeiene e N R 481097-0 1 US004 6 

### Test Order/DCI # EDSP-10990 1 -15 6



# if ) Title : 150-DAYRESPONS E TO ERA' S TES TORDE R FO R ENDOCRIN E SCREENIN G O N TRIADIMEFO N



### Guideline : OPPT S 89 0 Guidelin e Serie s

Authors : An n M . Blacker , PhD , DABT Tilghma n Hall , Ph D Patrici a Kwiatkowski , Ph D

Date : May25 , 201 0 Repor t ID : US004 6 

### Bayer CropScienc e L P P.O . Box1201 4 Researc h Triangl e Park , N C 2770 9

Pag e 1 o f4 6

## (iropScienc e US004 6 STATEMEN TO FN O DAT ACONFIDENTIALIT Y N o clai mo f

confidentialit y i s mad efo r an y informatio ncontaine di nthi s stud y o nth ebasi s o f it s fallin g withi nth e scop eo f FIFR A10(d)(l)(A) , (B ) o r (C) . Submissio no f

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o f an y righ t t o confidentialit y whic hma y exis t unde r an y othe r statut ei nan y countr y othe r tha nth eUSA . Th eabov estatemen t

supersede s al l othe r

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elsewhere i nthi s report . Company : Bayer Environmenta l Scienc e ADivisio no f

Baye r CropScienc eL P Compan y Agent : Date :

Richar dM . Gorrell , Ph.D . Registratio nProduc t

Manage r<NJttSrllM UI * US004 6

GOO D LABORATORYPRACTIC E COMPLIANC E STATEMEN T Thi s repor

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t t o th e principle s of4 0 CF R 160 , Goo d Laborator yPractice s Standard s (FIFRA) , as promulgate d i n Federa l Register , fina l rule . August 17.1989 .

5 / An n M . Blacker . PhD , DABTDat e Toxicdogist 

### Baye rCropScienc

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TilghmartWall , Ph D Date Ecotoxicologis

t Baye rCropScienc

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Patrici a Kwiat l Produc t Safet y Baye rCropScienc

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Richar dM . Gorrel l Date Manager , Registration s Baye r Environmenta l Scienc e ADivisio no f

Bayer CropScienc e LP Sponsor/Submitte r Pag e 3 o f4 6

## Bayer

CropScienc e US004 6 150-DA YRESPONS E T O ERA' S TES TORDE R FO R ENDOCRIN E SCREENIN G O N TRIADIMEFO N Introduction : O n Decembe r17 , 2009 , EP Aissue d a Tes t Orde rfo rendocrin e screenin g o n triadimefon . I n thi s Order , EP Aprovide s th e optio n t o cit e o rsubmi t existin g dat a t o satisf ypar t o ral l o fth e testing . Specifically , EP Astates,".. . suc h hazard-relate d dat a woul d b e o fhig h qualityan d achieve s th e objectiv e o fTie r1 assay s t o provid e reasonabl e assuranc e that a chemica l doe s o rdoe s no t have th e potentia l t o interact wit h th e estrogen , androgen , o rthyroi d systems. " Th e Triadimefo n Consortiu m wa s establishe d t o addres s EPA' s dat a call-i n an d include s th e followin g manufacturers : Baye rCropScienc e an d Tid e International . Studie s conducte d fo rregulator ypurpose s a s wel l a s studie s publishe d i n th e ope n literatur e wer e identifie d an d reviewe d b yBaye rCropScienc e o n behal fo fTh e Consortium . A n objectiv e assessmen t o fthes e studie s wa s performe d t o determin e whether , a s a bod yo fdata , the y provid e dat a functionall yequivalen t t o that i n EPA' s EDS P Tie r1 screenin g batter yan d includ e sufficien t informatio n t o characteriz e potentia l endocrine-modulatin g effect s o n estrogen , androge n o rthyroi d hormon e pathways . Whe n evaluatin g informatio n reporte d i n th e literature , i n general , th e approac h o fKlimisc h e t al . (1997 ) ha s bee n utilize d t o systematicall yasses s th e qualit yofwor k reported . Thi s approac h i s recognize d an d use d byregulator yauthoritie s includin g it s us e i n Europ e i n developin g th e Internationa l Unifor m Chemica l Informatio n Databas e (IUCLID) . Th e dat a reliabilitycriteri a presente d b yKlimisc h e t al . are simila rt o th e criteri a recommende d b yEP Ai n th e Hig h Productio n Volum e Progra m unde rTSCA . Assay s reporte d i n th e literatur e have als o bee n compare d t o th e relevan t assa ydescribe d i n th e OPPT S 89 0 Guidelin e Serie s t o identifymajo rdifference s betwee n th e specifi cassa yinclude d i n EPA' s validatio n progra m fo rTie r1 screenin g an d th e on e reporte d i n th e literature . Th e Triadimefo n Consortiu m i s citin g multipl e studie s an d publication s a s "Othe rScientificall y Relevan t Information " o rOSRI , whic h provid e dat a tha t ar e functionall yequivalen t to , an d effectivel ysubstitute s for , th e informatio n tha t woul d b e develope d throug h man yo fth e EDS P Tie r1 screenin g assays . W e believe tha t th e specifie d data , i n conjunctio n wit h conduct o ftw o Tie r1 assays , wil l provid e a tes t batter ytha t conserve s anima l usage , use s existin g informatio n appropriately , an d wil l adequatel yidentif yan ypotentia l endocrin e modulatin g effect s o f triadimefon . Muc h o fth e existin g dat a come s fro m U S EP Apublishe d researc h studie s an d guidelin e o rguidelin e equivalen t mammalia n toxicolog yregulator ystudie s whic h have bee n reviewe d b yEP Aan d hav e bee n classifie d a s acceptabl e i n th e Triadimefo n RE D an d subsequen t ris k

assessmen t documents . Endocrine-relate d apica l endpoint s whic h hav e bee n assesse d i n th e guidelin e studie s o n triadimefo n tha t ar e functionall yequivalen t t o endpoint s assesse d i n EPA' s Tie r1 endocrin e screenin g batter yare summarize d i n Tabl e 1 a & b . Functionall yequivalent , i n thi s context , refers t o dat a o fa suitabl e nature an d qualityt o provid e th e sam e essentia l predictiv e informatio n o n a chemica l eve n i fdifferen t method s an d procedure s mayb e use d fo robtainin g th e data . Further , th e majorityo fth e endpoint s require d i n th e puberta l assay s have bee n assessed , takin g int o accoun t bot h existin g guidelin e studie s an d relevan t finding s recentl yreporte d b yEP Aresearc h staf fi n th e ope n literature a s show n i n Tabl e 2 an d Tabl e 3 . Additionally , ther e i s a significan t bod yo fdat a o n thyroi d functio n an d histopathology , muc h o fi t fro m EP Aresearc h publications , t o serv e a s a n adequat e surrogat e fo rth e amphibia n metamorphosi s assa ya s show n i n Tabl e 4 . Pag e 4 o f4 6

## Bayer

CropScienc e US004 6 Th e Triadimefo n Consortiu m believe s tha t existin g data , a s reporte d i n bot h guidelin e studie s an d researc h publications , ar e fo rth e mos t par t adequat e t o asses s th e potentia l fo rthi s compoun d t o interact wit h estrogen , androge n an d thyroi d systems . Th e Triadimefo n Consortiu m believe s tha t conductin g th e followin g tw o assay s fro m th e EDS P Tie r1 screenin g batter ywil l provid e complementar ydat a an d sufficien t scientifi credundancyt o assur e tha t potentia l advers e endocrin e effect s woul d b e identifie d an d sufficien t dat a are availabl e t o perfor m a weigh t o fevidenc e (WOE ) evaluatio n o fwhethe ran yTie r2 testin g woul d b e necessary : I n viv

o assays : Fis h short-ter m reproductio n tes t

Hershberge rbioassay Th e principl e o fth e fis h short-ter m reproductio n scree n i s t o measure th e reproductiv e performanc e o ffathea d minnows a s th e primar yindicato rfo rendocrin e disruptio n (EP A200 9 890.1350) . Th e assa yscreen s fo rdisturbance s i n th e hypothalamic-pituitary-gonada l (HPG ) axi s includin g (anti- ) estrogenic , (anti- ) androgenic , aromatas e inhibition , steroi d modulatio n effects , examinin g abnormalitie s associate d wit h survival , reproductiv e behavior , secondar yse x characteristics , histopathology , an d fecundity(i.e. , numbe ro fspawns , numbe ro feggs/spawn , fertility , an d developmen t o foffspring ) offis h expose d t o tes t chemical s (USEP AOrder/DC I #EDSP-041101-42) . Equivalen t endpoint s are not availabl e wit h i n vivo organism s i n eithe r existin g guidelin e ecotoxicologica l studie s fo rtriadimefo n o ri n researc h studie s reporte d i n th e publi cdomain , an d therefor e th e Consortiu m wil l conduct th e fis h short-ter m reproductio n assay . Th e Hershberge rassa yi s a short-ter m i n viv o assa yt o detect androgeni co rantiandrogeni c chemical s o rchemical s tha t inhibi t 5a-reductas e b ymeasurin g weigh t change s i n fiv e androge n dependen t tissues : ventra l prostat e (VP) , semina l vesicl e (SV ) (plu s fluid s an d coagulatin g glands) , levato rani-bulbocavernosu s (LABC ) muscle , paire d Cowper' s gland s (COW ) an d th e glan s peni s (GP) . Sufficien t dat a o n thes e specifi ctissue s are no t availabl e i n guidelin e o ri n researc h studie s reporte d i n th e publi cdomain , an d therefor e th e Consortiu m wil l conduct th e Hershberge rassay . Followin g are assessment s o fth e existin g OSR I an d Th e Triadimefo n Consortium' s positio n o n thei rsufficiencyt o mee t th e requirement s o fth e variou s endocrin e screenin g assay s othe rtha n th e fis h short-ter m reproductio n assa yan d Hershberge rassay , beginnin g wit h a summar yo fth e guidelin e studie s bein g cited . Summar yo fTriadimefo n Guidelin e Studie s Al l studie s summarize d belo whav e bee n reviewe d b yU S EP Aan d judge d acceptabl e (Triadimefon . Preliminar

yHuman Health Ris k

Assessmen t (Revised) , D P Barcod e D326678 . Februar y9 , 200 6 an d Triadimefon : Human Health Ris k Assessmen t to Suppor

t Reinstatemen t ofUs e o n Residentia l Turf , Decisio n No . 391164 . Jul y24 , 2009) . Relevan t dat a whic h ar e functionall yequivalen t t o addres s th e purpos e o fth e variou s endocrin e relate d assay s i n EPA' s Tie r1 screenin g batter yar e availabl e fro m guidelin e studie s includin g tw o ra t reproductio n studies , a developmenta l neurotoxitystudy , developmenta l toxicitystudies , subchroni ctoxicitystudie s i n th e rat an d dog , lifetim e feedin g studie s i n rat s an d mic e an d a 2 Pag e 5 o f4 6

# f

A

# t« 4 Bayer CropScienc e US004 6 yea rchroni cdo g toxicit ystudy . Functionall yequivalent , i n thi s context , refers t o dat a o fa suitabl e nature an d qualityt o provid e th e sam e essentia l predictiv e informatio n o n a chemica l eve n i fdifferen t method s an d procedure s mayb e use d fo robtainin g th e data . I n th e firs t multigeneratio n reproductiv e toxicit ystud ywit h triadimefo n (Loser , E. , Lorke , D. , 1979 ; MRI D 32541 ) mal e an d femal e rat s wer e administere d triadimefo n continuousl yi n th e die t at concentration s o f0 , 50 , 30 0 o r180 0 pp m an d th e reproductiv e performanc e studie d ove r3 consecutiv e generations , eac h wit h 2 matings . Offsprin g o feac h firs t matin g (F1a , 2 a an d 3a ) wer e sacrifice d o n PN D 28 ; offsprin g fro m th e secon d mating s wer e use d fo rsubsequen t mating s (F1b , 2 b receive d treate d die t fro m weanin g unti l approximatel yPN D 10 0 an d the n wer e mated , wit h female s continuin g t o receiv e treate d die t throug h gestatio n an d lactation ) o r offsprin g histopatholog yinvestigatio n (F3b) . I n a complementar ytwo-generatio n reproductive toxicitystud y(Eiben , R. , 1984 ; MRI D 155075 , 9218801 9 & 92188320) , triadimefo n wa s administere d continuousl yi n th e die t t o rat s i n th e die t at level s o f0 , 5 0 o r180 0 pp m a s follows : P animal s wer e give n tes t articl e die t formulation s fro m 10 0 day s prio rt o matin g t o produc e th e F 1 generation . Afte rweaning , F 1 animal s wer e selecte d t o becom e th e F 1 parent s o fth e F 2 generatio n an d wer e give n th e sam e concentratio n tes t formulatio n followin g th e sam e patter n a s thei rdam s excep t that onl yth e 5 0 pp m F 1 parent s wer e sacrifice d whe n th e F 2 litters wer e weaned . Treatmen t o fth e F 1 hig h dos e group s wa s stoppe d an d th e hig h dos e F 1 male s wer e mate d wit h th e contro l F 1 females , whil e th e hig h dos e F 1 female s wer e mate d wit h contro l F 1 males . Thi s cross-matin g procedur e produce d a secon d F 2 litte ran d wa s intende d t o provid e furthe rinformatio n regardin g th e fertilityo fth e F 1 parents . Fo ral l matings , th e daytha t sper m wa s detecte d i n a vagina l smea rwa s designate d a s gestatio n day(GD ) 0 . Sper m motility , sper m morphology , gestatio n length , an d se xratio s wer e unaffecte d b ytreatment . Whe n administere d at level s u p t o 30 0 ppm , triadimefo n ha d n o effect o n mating , fertility , pregnancyo rlactatio n performanc e o fPO , F 1 o rF 2 females . There wer e n o advers e effect s o n pu p weight s at birt h o rsurviva l rate s durin g lacatio n i n F1 , F 2 o rF 3 pups . I n th e firs t study , th e dietar yleve l o f30 0 pp m ha d sligh t effect s o n F2 b an d F3 b pu p bod yweight s durin g lactation . N o histopathologica l change s wer e observe d i n an yorga n examine d (eyes , adrenals , brain , thyroid , thymus , heart , lung , liver , spleen , pancreas , lymp h nodes , stomach , intestine , kidneys , testes , epididymis , ovaries , uterus ) fro m fou rwee k ol d F3 b pup s whic h wer e euthanize d an d necropsied . I n bot h studies , at 180 0 pp m fertilitywa s affecte d i n al l generations . Bod yweight s ofP , F 1 an d F 2 generatio n parent s wer e decrease d durin g premating , mating , gestation , an d lactation . Bod y weigh t gain s o fth e F 1 generatio n parenta l male s an d female s wer e als o decrease d throughou t pre-mating , gestation , an d lactatio n fo rbot h matings . I n th e firs t study , th e numbe ro foffsprin g pe rda m at birt h wa s decrease d i n th e firs t litters o fbot h th e P generatio n an d th e F 1 generation . Fertilityvalue s wer e significantl ylowe ri n bot h mating s o fth e F 1 generation . Onl y on e femal e becam e pregnan t i n th e F2 a mating , an d n o female s becam e pregnan t i n th e F2 b mating . I n th e secon d study , at 180 0 ppm , fertilitywa s als o significantl ydecrease d i n th e F 1 parents . Additionally , bod yweight s wer e decrease d i n th e F 1 parenta l male s an d female s durin g week s 9-4 0 an d bod yweight s wer e decrease d at birt h an d durin g lactatio n i n th e F 1 an d F 2 pups . F 2 litte rsiz e wa s decreased ; viabilitywa s decrease d i n th e F 1 an d F 2 pup s durin g lactation . Elevate d seru m testosteron e level s wer e reporte d fo rhig h dos e F 1 male s at necropsy , bu t luteinizin g hormon e an d estradio l level s wer e unaffected . N o histopathologica l change s Pag e 6 o f4 6

# Bayer

CropScienc e US004 6 wer e observe d i n an yreproductiv e organ s examine d (testes , ovaries , epididymis , pituitary , prostate , semina l vesicles , uteru s an d vagina ) fro m necropsie d F 1 parent s o rF 2 pups . I n th e F 1 cross-matin g (secon d study) , fertilitywa s 47.4 % fro m th e matin g o fhig h dos e male s wit h th e contro l female s an d 80 % fro m th e matin g o fth e hig h dos e female s wit h th e contro l males . I n th e absenc e o fsper m abnormalities , i t wa s conclude d tha t th e observe d decreas e i n fertilitywa s du e t o a decrease d interes t o fth e male s i n copulatio n (copulatio n wa s no t observe d fo r6/1 0 o fth e 180 0 pp m male s durin g th e 3-wee k cross-matin g wit h contro l females) . I n a developmenta l neurotoxicitystud ywit h triadimefo n (Sheets , L , e t al. , 2008 ; MRI D 47377101) , pregnan t rat s wer e administere d triadimefo n i n th e die t at level s o f0 , 100 , 30 0 o r 80 0 pp m fro m G D 6 t o lactatio n day(LD ) 2 1 whe n pup s wer e weane d an d pu t ont o untreate d diet . Offsprin g wer e evaluate d fo ra varietyo fparameters includin g bod yweight , acquisitio n o f vagina l patenc

yan d preputia l separation , surfac e righting , pupi l constriction , acousti cstartle , moto ractivityan d learnin g an d memory . Neura l tissue s wer e take n at post nata l day(PND ) 2 1 an d 7 5 fo rmicroscopi cexaminatio n an d morphometry . Parenta l reproductio n parameters , litte rsiz e an d viabilitywer e unaffecte d b ytreatment . Th e acquisitio n o fvagina l patenc yan d preputia l separatio n i n F 1 offsprin g wa s unaffecte d at materna l dietar

ylevel s u p t o 80 0 ppm , th e highes t dos e tested ; offsprin g surfac e rightin g an d pupi l constrictio n wer e als o unaffected . Moto ran d locomoto ractivit ywa s unaffecte d fo rbot h sexe s i n al l dos e groups . At 80 0 ppm , ther e wer e decrease s i n pre-weanin g bod yweigh t (PN D 11 , 17 , an d 21 ) i n bot h sexes , an d pre-weanin g bod yweigh t gai n i n bot h sexes ; deviate d snou t wa s see n i n 3 female s an d 1 male . A n increase d pea k amplitud e durin g auditor ystartl e refle x testin g wa s observe d i n hig h dos e female s onl yo n PN D 6 0 an d increase d numbe ro ftrial s t o criterio n durin g th e retentio n phas e o fth e avoidanc e tes t i n female s a s well . Ther e wer e n o difference s i n micropatholog ybrai n measurement s i n day2 1 o r7 5 animal s considere d t o b e treatmen t related . I n developmenta l toxicit ystudie s investigatin g th e effect o ftriadimefo n o n th e developin g fetu s i n rats , ther e wer e n e effect s o n th e numbe ro fcorpora lutea/dam , implants/dam , viabl e fetuses , earl yo rlat e res«rptions , non-viabl e implantations/dam , live litters , fetuses/da m o ro n se xrati o at materna l d»se s u p t o 9 0 mg/k g bw/dayi n on e stud y(Linger , T . e t al.,1982 ; MRID s 14933 6 & 92188018 ) an d 10 0 mg/kg bw/da yi n th e othe r(Nagumo , K . e t al. , 1981 ; MRI D 89023 ) eve n i n th e presenc e o fsignifican t materna l toxicitya s evidence d b ydecrease d bod y weigh t gain s an d increase d moto ractivity . I n a rabbi t developmenta l toxicitystud y(Clemens , G . an d Hartnagel , R. , 1990 ; MRI D 4144620 1 & 42089601 ) wit h triadimefo n ther e wa s als o n o treatment-relate d effect o n gestatio n o rfertilityindice s at materna l dose s u p t o 12 0 mg/kg bw/day . Ther e wer e n o overt sign s o ftoxicit yi n subchroni crat (Mohr , U. , 1976 ; MRI D 4862 4 ) an d subchroni cdo g (Hoffman , K . an d Luckhaus , G. , 1974 ; MRI D 4862 5 & 60226 ) toxicit ystudie s at th e highes t dose s teste d (10 0 an d 6 0 mg/k g bw/day , respectively) . Ther e wer e n o treatmen t relate d effect s note d o n adrenals , thyroid , pituitary , ovar yo rtesti s tissue s examine d a t th e highes t dose s tested . I n 2 1 mont h mous e oncogenicit y(Bomhard , E. , 1986 ; MRI D 4075210 1 & 4086510 1 ) , 2 4 mont h rat toxicity/oncogenicit y(Bomhard , E . an d Schilde , B. , 1991 ; MRI D 42153901 ) an d 2 4 Pag e 7 o f4 6

## Baye r

CropScienc e US004 6 mont h do g toxicit y(Hoffman , K . an d Groning , P. , 1978 ; MRI D 0003253 9 & 00126261 ) studies , ther e wer e n o treatmen t relate d effect s note d o n an yreproductiv e orga n examine d at an ydos e leve l (highes t dose s tested : mouse , 76 5 an d 55 0 mg/k g bw/da yfo rfemale s an d males , respectively ; rat , 19 9 an d 11 4 mg/k g bw/da yfo rfemale s an d males , respectively ; dog , 68. 8 an d 60. 4 mg/k g bw/dayfo rfemale s an d males , respectively) . I n th e mous e oncogenicitystudy , th e LOAE Li s 119/7 6 mg/k g bw/day(f/m) , th e middl e dos e tested , base d o n live rhypertroph yi n bot h sexe s an d othe rse xspecifi clive ranomolies , wit h a treatmen t relate d increas e i n heptacellula radenomas . I n th e rat combine d chroni c toxicity/oncogencit ystudy , th e LOAE Li s equivalen t t o 199/11 4 mg/kg bw/da y(f/m) , th e highes t dos e tested , base d on : increase d foo d consumptio n an d incidence s o ffat i n hepatocyte s i n bot h sexes ; increase d alanin e aminotransferas e level s i n males ; increase d absolut e an d relativ e live r weigh t an d cholestero l level s i n female s an d decrease d bod yweigh t an d bod yweigh t gain s i n females . Th e carcinogeni cpotentia l o ftriadimefo n i n rat s wa s considere d equivoca l b yU S EP A base d o n th e incidenc e o fthyroi d adenoma s an d cysti chyperplasi a i n bot h sexes , whic h wer e suggeste d byBC S t o b e withi n historica l contro l rang e (Sander , E . an d Schilde , B. , 1993 ; MRI D 43089301) . Fo rth e 2 yea rdo g study , th e LOAE Li s 34.70/33.6 7 mg/k g bw/day ( m/f) , th e middl e dos e tested , base d o n increase d alkalin e phosphatas e level s i n bot h sexes ; an d decrease d foo d consumptio n an d increase d cholestero l i n females . Th e Cance rAssessmen t Revie wCommitte e (CARC ) classifie d triadimefo n into categor yC "possibl e huma n carcinogen " - base d o n a statisticall ysignifican t increas e i n thyroi d adenoma s i n mal e Wista rrat s an d statisticall ysignifican t increase s i n hepatocellula radenoma s i n bot h sexe s o fth e NMR I mouse . However , i t wa s conclude d tha t a quantifie d carcinogeni cris k assessmen t fo rtriadimefo n i s no t appropriat e an d ris k assessmen t wil l b e base d o n th e cPA D an d Margi n o fExposure approac h only . Othe rScientificall yRelevan t Informatio n Estroge n Syste m OSR I
» Mammalia n assay s (estroge n recepto r(ER ) binding , E R transcriptiona l activation , uterotrophic , puberta l female ) have bee n include d i n th e EDS P Tie r1 batter yt o asses s th e potentia l o fa chemica l t o affec t th e estroge n signalin g pathway . Non e o fth e assay s a s specificall ydescribe d i n EPA' s OPPT S 89 0 Guidelin e Serie s have bee n conducte d wit h triadimefon . However , a s show n i n Table s 1 a & 1 b , dat a relevan t t o E R bindin g an d transcriptiona l activation , uterotrophic , an d puberta l femal e assays are availabl e fro m guidelin e studies , al l ofwhic h hav e bee n reviewe d b yEP Aan d classifie d a s acceptable . Estrogeni can d anti-estrogeni cactivitie s hav e bee n examine d independentl yb ytw o researc h group s i n a n MCF- 7 cel l proliferatio n assa y (Vinggaard e t al. , 1999 ; Okub o e t al. , 2004) . I n addition , triadimefo n wa s include d i n a Japanes e investigatio n that screene d 20 0 pesticid e chemical s fo restroge n an d androge n activitie s (Kojim a e t al. , 2004) . Th e thre e publication s relevan t t o E R activityare detaile d below . Also , dat a fro m th e tw o regulator ymulti-generatio n rat reproductio n studie s an d relevan t result s fro m a recen t publicatio n authore d b yU S EP Ascientist s reportin g effect s o fconazol e fungicide s o n reproductiv e developmen t i n femal e rat s (Rockett e t al. , 2006 ) are presente d t o adequatel y addres s th e requiremen t fo ra femal e puberta l assa y(Tabl e 2) .

Pag e 8 o f4 6

# Baye r

CrqpScienc e US004 6 I n vitr o Estroge n Recepto rBindin g Assa yan d Estroge n Recepto rTransactivatio n Assa y Althoug h n o studie s have bee n conducte d accordin g t o th e 89 0 Guideline s issue d b yEPA , specificall yOPPT S 890.125 0 an d 890.1300 , estrogeni can d anti-estrogeni cactivitie s have bee n examine d independentl yb ytw o researc h group s i n a n MCF- 7 cel l proliferatio n assa y (Vinggaard e t al. , 1999 ; Okub o e t al. , 2004) . Arecombinan t yeas t strai n (Saccharomyce s cerevisaie) , stabl ytransfecte d wit h th e huma n estroge n recepto ra an d a reporte rplasmi d encodin g fo rth e enzym e (3-galactosidas e (originall ydevelope d b yGlaxo-Wellcome ) wa s als o use d b yVinggaar d t o examin e triadimefon' s estrogeni cactivity . Althoug h triadimefo n di d no t produc e an yrespons e i n th e yeas t screen , th e lac k o factivit ywa s attribute d t o cytotoxicity . I n bot h cel l proliferatio n experiments , huma n breas t cance restrogen-sensitiv e MCF- 7 cell s wer e expose d t o a serie s o ftriadimefo n concentration s (Vinggaard : 0.001 , 0.1, 1 an d 10uM , dissolve d i n medium ; Okubo : 6 concentration s rangin g fro m 1 t o 1000uM , dissolve d i n ethanol : combinin g bot h studies , a tota l o f8 concentration s wer e teste d rangin g fro m 10~ 9 t o 10~ 3M) . I n bot h experiments , 1 7 (3-estradio l wa s use d a s th e positiv e contro l an d referenc e t o calculat e relativ e proliferatio n potency(RPP ) an d relativ e proliferatio n efficiency/effect (RPE) . Th e Okub o investigatio n include d cytotoxicityinformation ; triadimefo n wa s reporte d no t t o b e cytotoxi cat concentration s u p t o 1000uM . I n bot h experiments , triadimefo n induce d a n increas e cel l proliferatio n an d simila rRPE s wer e reported . Bot h publication s identifie d triadimefo n a s a wea k estroge n recepto ragonist . Additiona l experiment s wer e conducte d an d reporte d i n th e Okub a e t al . publication . Th e effec t o fantiestroge n IC I 182,78 0 o n cel l proliferatio n induce d b ytriadimefo n wa s als o examine d a s wel l a s th e antiestrogeni cactivityo ftriadimefo n a s determine d b yth e suppressio n o fcel l proliferatio n induce d by3 0 p M 1 7 (3-estradiol . Additio n ofagonis t IC I 182,78 0 significantl y decrease d th e cell-proliferativ e effec t o ftriadimefo n t o approximatel yth e contro l (0.1 % ethanol ) level . I n th e antiestroge n assay , triadimefo n wa s antiestrogeni cat 10 0 an d 100 0 uM . Finally , th e bindin g affinityo ftriadimefo n fo rhuma n ER a o rA R wa s determine d wit h ligan d screenin g recepto rsystems . Ther e wa s s o littl e bindin g o ftriadimefo n t o ER a tha t a n IC5 0 valu e coul d no t b e calculated , an d th e IC5 0 valu e o ftriadimefo n t o A R wa s 90 2 ±14 3 u M wit h a n RB A(relativ e bindin g affinitycompare d t o positiv e contro l mibolerone ) o f0.00 5 (IC5 0 value s bein g th e concentratio n o fcompoun d tha t inhibit s bindin g o f1 7 (3-estradio l t o E R o rtha t o ftestosteron e t o A R b y50%) . Result s o fanothe rE R assa ywit h triadimefo n hav e bee n reporte d i n th e literature . Kojim a et . al . (2004 ) investigate d th e potentia l fo r20 0 pesticides , includin g triadimefon , t o bin d t o th e alph a an d bet a E R usin g transfecte d Chines e Hamste rOvary-K 1 cells . Triadimefo n wa s teste d at concentration s rangin g fro m ~10~ 8 t o 10" 5 M , an d th e REC2 0 (concentratio n o fchemica l tha t result s i n 20 % o fth e agonis t activit yo f10" 1 0 M E2 ) valu e an d relativ e estrogeni cactivit yat 10~ 5 M concentratio n (RLA ) wer e measured . Triadimefo n wa s no t reporte d t o sho westrogeni c activity . However , i t mus t b e note d tha t th e authors teste d fewe rtha n th e eigh t concentration s require d b yth e OPPT S 890.125 0 Guidelin e an d di d no t provid e informatio n o n solubilityo r cytotoxicity . Thus , thes e result s canno t b e considere d reliabl e o n thei rown , bu t are consisten t wit h th e conclusio n tha t estroge n recepto ractivatio n i s no t a n importan t mod e o factio n fo r triadimefon .

Pag e 9 o f4 6

## Bayer

CropScienc e US004 6 Anothe rstud yexamine d th e interactio n o ftriadimefo n wit h th e rainbo wtrou t estroge n receptor , usin g a recombinan t yeas t cel l line , a s wel l a s rainbo wtrou t hepatocyt e aggregat e culture s (Peti t at al . 1997) . Tes t concetration s fo rtriadimefo n (an d 4 9 othe rindepentl yteste d compounds ) range d fro m 10~ 8 t o 10" 4 M . Th e yeas t cel l lin e showe d simila rresult s a s abov e wit h n o activitywit h th e receptor , mos t likel yt o du e t o cel l death . Wit h th e trou t hepatocyt e cel l line , th e stud ydoe s no t provid e th e result s o fal l teste d concentrations , an d therefor e interpretatio n o fth e result s i n limited . However , i t appears tha t triadimefo n di d no t displac e estradio l bindin g t o th e trou t estroge n receptor , howeve rvitellogeni n inductio n wa s noted . Th e authors sugges t tha t tha t activity(vitellogeni n induction ) wa s du e t o eithe ra metabolit e o rt o a n indirec t gen e activiatio n pathway . Base d upo n th e consistenc yofth e finding s i n thre e independentl yconducte d i n vitr o E R assa y experiments , th e requiremen t fo rth e EDS P Tie r1 i n vitr o estroge n recepto rbindin g assa yan d estroge n recepto rtransactivatio n assa yshoul d b e waived . I n viv o Femal e Puberta l Assay : Th e effect s o ftriadimefo n o n th e reproductiv e developmen t i n th e femal e Wista rHa n rat wer e als o investigate d b yU S EP Aresearchers (Rockett e t al. , 2006) . Mate d femal e rat s wer e administere d triadimefo n vi a th e die t (100 , 500 , o r180 0 ppm ) fro m gestatio n day6 throug h lactation . Femal e offsprin g wer e expose d vi a lactatio n unti l PN D 2 2 an d the n receive d th e sam e dietar yconcentration s a s thei rdam s unti l PN D 98 . Thus , th e exposur e duratio n fo rth e F 1 female s exceed s tha t require d b yth e puberta l assa yan d therefor e provide s a more sever e assessmen t o fa compound' s potentia l t o affect endocrine-relate d endpoints . Bod yweigh t (BW ) an d anogenita l distanc e (AGO ) at PN D 0 , ag e an d B Wat vagina l openin g (VO) , estrou s cyclicity , an d bod yan d variou s orga n weight s a t necrops ywer e sai d t o b e measured . Liver , ovarie s an d thyroi d wer e examine d microscopically . Triadimefo n administere d at 50 0 pp m i n th e die t ha d n o significan t advers e effec t o n dam s o r offspring . Offsprin g anogenita l distanc e o n dayo fbirt h wa s unaffecte d at al l treatmen t levels ; V O wa s comparabl e t o control s fo rlo wan d mid-dos e female s (consisten t wit h th e finding s i n th e triadimefo n developmenta l neurotoxicitystud y(Sheets , L , 2008 ) wher e n o effec t o n V O i n female s o rpre-putia l separatio n (PPS ) i n male s wa s note d i n pup s bor n o fmothers receivin g triadimefo n level s at u p t o 80 0 pp m i n th e die t fro m GD 6 t o LD21) . Th e reporte d dela yi n V O i n hig h dos e female s i s confounde d b yth e absenc e o fbod yweigh t dat a i n th e publication . Th e authors stat e tha t bod yweight s a t birt h an d V O wer e unaffecte d b ytriadimefo n bu t a t necrops y (PN D 99 ) weight s o fhig h dos e animal s wer e significantl ylowe rtha n th e controls . I t seem s improbabl e tha t th e developin g pu p bod yweight s fo rth e hig h dos e grou p wer e unaffecte d b y treatment , give n bot h th e statisticall ysignifican t decreas e i n foo d consumptio n fro m weanin g t o necrops ya s reporte d i n th e publicatio n detaile d i n Tabl e 3 (i n particula rfro m PN D 29-43 , p<0.001 ) i n additio n t o th e reporte d statisticall ysignifican t reduce d bod yweight s (p<0.001PND1 , 8 an d <0.000 1 PN D 2 2 t o 92 ) reporte d fo rhig h dos e mal e pup s i n th e mal e cohor t stud y(Goet z et.al . 2007) , whic h i s summarize d later . I n addition , a s presente d earlier , i n bot h regulator

ymulti-generatio n reproductio n studies , adul t rat s receivin g 180 0 pp m o f triadimefo n i n thei rdie t a s wel l a s offsprin g ha d significantl yreduce d bod yweight s ove rmos t i f no t al l o fthei rlif e spans . Therefor e th e significanc e o fth e reporte d V O delayi n hig h dos e offsprin g i s questionabl e i n th e absenc e o fbod yweigh t data . Further , i n th e Rockett publication , relativ e live ran d ovaryweight s wer e reporte d increase d i n femal e offsprin g at th e hig h dose . Pag e 1 0 of4 6

## Baye r

CropScienc e US004 6 Onl ymea n relativ e orga n weight s are mentione d i n th e repor t whe n discussin g significan t difference s betwee n hig h dos e animal s an d control ; absolute orga n weight s an d bod yweights ar e neede d t o understan d th e toxicologica l significanc e o fchange s i n relativ e orga n weights . Histopatholog yshowe d live ran d thyroi d effect s considere d t o b e treatmen t related . Ther e were n o unusua l observation s fo rovaries ; al l ovarie s containe d follicle s at al l stage s o fdevelopment , ha d norma l appearin g corpor a lute a an d showe d n o sign s o fedema , cysti cfollicle s o rretaine d oocytes . I n th e tw o multi-generatio n rat studie s (Lose ran d Lorke , 1979 : Eiben , 1984 ) P adult s wer e expose d t o triadimefo n i n th e die t fo rat leas t 1 0 week s prio rt o matin g an d the n throughou t mating , gestation , an d lactatio n periods . Indirec t exposur e fo rth e F 1 offsprin g covers th e i n uter o an d lactatio n period s wit h direct exposur e startin g aroun d PN D 1 1 o r1 2 whe n pup s begi n t o consum e treate d die t an d fro m PN D 28 , continue s fo rat leas t 1 0 week s prematin g throughou t mating , gestation , an d lactatio n period s fo rpup s selecte d t o becom e F 1 parents . Thus , th e exposur e duratio n fo rth e F 1 generatio n exceed s tha t require d b yth e puberta l assa yan d therefor e provide s a more severe assessmen t o fa compound' s potentia l t o affec t endocrine relate d endpoints . Th e result s o fthes e studie s hav e bee n discusse d above . Ther e wer e n o fertilityo rse xorga n relate d finding s i n eithe rmale s o rfemale s tha t sugges t estroge n disruption . Th e puberta l assa yi s designe d t o detect effect s o n puberta l developmen t an d thyroi d function . As detaile d i n Tabl e 2 , al l ofth e sexua l developmenta l endpoints include d i n a femal e puberta l assa yhav e bee n assesse d fo rtriadimefon . Wit h th e exceptio n o fclinica l chemistryan d thyroi d hormon e measurements , th e endpoint s i n th e femal e puberta l assa yare capture d i n th e "highe r tiered " two-generatio n rat reproductio n studie s an d researc h result s presente d b yRockett e t al . (2006) . Complet e clinica l chemistr ydat a are availabl e fo rmultipl e timepoint s i n th e chroni crat an d do g studies . Thyroi d weigh t an d histolog yare availabl e fro m chroni cstudie s i n rats , mic e an d dog s a s discusse d earlier . Thyroi d hormon e leve l measurement s ar e no t availabl e fo r femal e rat s bu t hav e bee n measure d i n mal e rat s b yEP Aresearchers an d wil l b e discusse d late ri n relatio n t o th e androge n assays . Th e result s fro m al l o fthes e studies , i n particula rth e two-generatio n rat reproductio n studies , developmenta l neurotoxicitystud yan d 200 6 publicatio n b yRockett , e t al. , shoul d b e considere d adequat e t o addres s th e dat a requiremen t fo ra femal e puberta l assay . Therefore , th e nee d t o conduct thi s assa yshoul d b e waived . I n viv o Uterotrophi cAssa y EP Astate s tha t uterotrophi crespons e i s no t du e exclusivel yt o estrogeni cchemical s s o that a Uterotrophi crespons e shoul d b e confirme d b ycorroboratin g informatio n suc h a s E R bindin g o r transcriptiona l activation . Finding s i n publishe d transcriptiona l activatio n studie s di d no t sugges t tha t estroge n recepto ractivatio n i s a n importan t mod e o factio n fo rtriadimefon . I n addition , neithe rth e multi-generatio n reproductio n studie s no rth e researc h byRockett et al . suggest estrogeni cactivity . Therefore , th e nee d t o conduct thi s assa yshoul d b e waived . Conclusio n fo rE R Assay s Advers e effect s tha t woul d b e expecte d fro m exposur e t o a n estroge n disruptin g chemica l ar e show n below . Non e o fth e effect s are consisten t wit h wha t ha s bee n observe d i n mammalia n toxicolog ystudie s wit h triadimefon . Pag e 1 1 of4 6

# p A 

# tE 

# 4 Baye r

CropScienc e US004 6 Mechanis m Potentia l Effect s i n Male s Potentia l Effect s i n Female s Estrogenicity Delaye d preputia l separatio n (marked , o ri n absenc e ofsignifican t bod yweigh t decreases ) Reduce d fertility Reduce d sper m count s Decrease d mal e reproductiv e orga n weight s (excep t prostat e weight , whic h mayb e increased ) Histopathologica l finding s o fth e teste s (e.g. , Leydi g cel l proliferatio n and/o rtumors ) Increase d incidence , growt h of pituitar ytumors Precociou s vagina l openin g Persisten t estru s Increase d tim e t o matin g Reduce d gestatio n duratio n Increase d uterin e weight s Histopathologica l finding s o fth e femal e reproductiv e organ s (e.g. , vagina l cornification , uterin e hypertroph yan d hyperplasia ) Increase d mammar

ytumors (adenoma s o radenocarcinomas ) Increase d incidence , growt h o f pituitar ytumor s Ovarian , uterin e an d vagina l tumors i n femal e offsprin g Anti-estrogenicity Decrease d epitheliu m heigh t i n testicula rtubule s Testicula rweigh t increase s (shor t term ) Testicula ratroph y (fro m long-ter m exposure ) Infertilityan d testicula ratroph y (fro m moderate-ter m exposures ) Delaye d vagina l openin g Delaye d star

t o festrou s cycling ; irregula ro rabsen t estrou s cyclicity Reduce d fertility Decrease d corpora lutea , implantation s Decrease d femal e reproductiv e orga n weight s Decrease d mammar

ytumo r incidenc e I n summary , scientifi cinformatio n indicate s tha t a n estrogeni co ranti-estrogeni ccompoun d woul d produc e amon g othe rfinding s disturbance s i n mal e fertility/sper m coun t o rmotility , estrou s cyclicit

yand/o ralteration s i n th e weigh t an d histopatholog yofspecifi creproductiv e organ s (e.g. , ovary , uterus , testis , o repididymis) . Adequat e informatio n o n triadimefo n effect s o n al l parameters i s availabl e i n existin g regulator ystudie s an d publication s authore d b yEP A researchers . Give n th e leve l o favailabl e information , i t i s considere d unnecessar yt o conduct eithe ro fth e i n vitr

o estroge n syste m assay s o rt o sacrific e th e additiona l animal s neede d t o conduct th e femal e puberta l assa yo rth e uterotrophi cscreenin g assay . Therefore , th e requiremen t fo rth e estrogen-base d EDS P Tie r1 mammalia n assay s shoul d b e waived . Whil e th e rainbo wtrou t hepatocyt e cel l line s mayhav e demonstrate d a n inductio n o f vitellogeni n (Peti t e t al . 1997) , th e mechanis m fo rsuc h inductio n wa s no t clear . Fo rthi s reason , th e Triadimefo n Consortiu m ha s agree d t o conduct th e fis h short-ter m reproductio n test . Androge n Syste m OSR I Mammalia n assay s (androge n recepto r(AR ) binding , Hershberger , puberta l male ) have bee n include d i n th e EDS P Tie r1 batter yt o asses s th e potentia l o fa chemica l t o affec t th e androge n signalin g pathway . Non e o fth e assay s a s specificall ydescribe d i n EPA' s OPPT S 89 0 Guidelin e Serie s hav e bee n conducte d wit h triadimefon . However , a s show n i n Table s 1 a & 1 b , dat a relevan t t o A R binding , Hershberge ran d puberta l mal e assay s ar e availabl e fro m guidelin e studies , al l o fwhic h hav e bee n reviewe d b yEP Aan d classifie d a s acceptable . Androgeni c activityha s bee n examine d wit h a ligan d screenin g androge n recepto rsyste m b yOkub o e t al. , Pag e 1 2 o

f4 6

# Bayer

CropScienc e US004 6 200 4 a s reporte d above . I n addition , triadimefo n wa s include d i n a Japanes e investigatio n tha t screene d 20 0 pesticid e chemical s fo restroge n an d androge n activitie s (Kojim a e t al. , 2004) . Also , dat a fro m th e tw o regulator ymulti-generatio n ra t reproductio n studie s an d relevan t result s fro m a recen t publicatio n authore d b yU S EP Ascientist s reportin g o n th e reproductiv e toxicit yo f conazol e fungicide s i n mal e rat s (Goet z e t al. , 2007 ) are presente d t o adequatel yaddres s th e requiremen t fo ra mal e puberta l assa y(Tabl e 3) . I n vitr o Androge n Bindin g Assa y A s reporte d above , bot h Okub a e t al. , 200 6 an d Kojim a e t al. , 200 4 include d triadimefo n i n androge n screenin g assays . Althoug h neithe rwa s conducte d accordin g t o th e EP Aprotoco l fo r th e Tie r1 assay , th e fact that bot h assay s showe d essentiall yn o bindin g shoul d b e sufficien t evidenc e tha t triadimefo n doe s no t act at th e androge n receptor . I n viv o Mal e Puberta l Assa y Th e reproductiv e toxicit yo ftriadimefo n o n i n th e mal e Wista rHa n rat wa s investigate d b yU S EP Aresearchers (Goet z e t al. , 2007) . Mate d femal e rat s wer e administere d triadimefo n vi a th e die t (100 , 500 , o r180 0 ppm ) fro m gestatio n day6 throug h lactation . Mal e offsprin g wer e expose d vi a lactatio n unti l PN D 2 2 an d the n receive d th e sam e dietar yconcentration s a s thei r dam s unti l PN D 92 . Thus , th e exposur e duratio n fo rth e F 1 male s exceed s tha t require d b yth e puberta l assa yan d therefor e provide s a more sever e assessmen t o fa compound' s potentia l t o affect endocrine-relate d endpoints . B Wan d AG O wer e measure d o n PNDO . Foo d consumptio n an d B Wwer e measure d weekly . Ag e an d B Wwer e recorde d o n th e dayo fPPS . On e mal e fro m eac h litte rwa s take n t o necropsyat PND1 , 22 , 5 0 an d 9 2 t o asses s effect s o n orga n weight s (brain , hypothalamus , pituitary , thyroid , liver , testis , epididymis , ventra l prostrate s an d semina l vesicles) , histolog y(brain , pituitary , thyroid , liver , testis , epididymi s an d ventra l prostrates ) an d hormon e measurement s (estradiol , testosterone , tota l triiodothyronin e (T3) , tota l thyroxin e (T4) , thyroi d stimulatin g hormon e (TSH ) an d luteinizin g hormon e (LH) . Du e t o th e limite d numbers o f hig h dos e male s survivin g t o weaning , a PND5 0 necrops ywa s no t don e fo rth e hig h dos e group . On e testi s an d on e epididymi s fro m eac h mal e at PND22 , 5 0 an d 9 2 necrops ywer e use d fo rsper m morpholog yan d motilityanalysis . Matin g assay s wer e conducte d o n PND7 8 o rolde r treate d male s wit h untreate d females . Ejaculate d sper m count s afte rnatura l breedin g wer e measure d fo rhig h dos e males . Ther e wer e n o treatmen t relate d effect s o n growt h o rsurviva l at 50 0 pp m an d below . Hig h dos e mal e offsprin g ha d reduce d bod yweigh t fro m birt h an d remaine d les s tha n contro l throughou t th e stud y(statisticall ysignifican t at al l times) . Surviva l rat e i n th e hig h dos e grou p wa s reduce d t o 47% ; ther e wer e n o death s i n animal s tha t survive d past PND8 . AG O wa s significantl y increase d i n male s i n th e 180 0 pp m group , an d PP S wa s delayed , a findin g confounde d b y reduce d pu p size/BW . Ther e wer e n o effect s note d i n mid - an d low-dos e pups . Sper m hea d an d tai l morpholog yan d sper m motilitywer e unaffecte d b ytreatmen t at al l levels . As observe d i n th e multigeneratio n studies , male s receivin g 180 0 pp m apparentl ydi d no t mate ; n o effect o n inseminatio n indexo rpercen t fertilitywas note d fo rmale s receivin g 10 0 o r50 0 ppm , wit h al l fertilityparameters i n th e pregnan t female s comparabl e t o controls . Elevate d seru m testosteron e level s wer e reporte d fo rhig h an d mid - male s at necropsy , bu t luteinizin g hormon e an d estradio l level s wer e unaffected , agai n consisten t wit h multi-generatio n stud y Pag e 1 3 of4 6

## Baye r

CropScienc e US004 6 findings . T4 level s i n 180 0 pp m male s wer e decrease d at PND9 2 onl ybu t neithe rT3 no rTS H level s wer e affected . Relative , bu t no t absolute , live rweight s wer e increase d i n PND 1 offsprin g at al l dos e levels , i n mid-dos e male s at PND5 0 an d onl yi n hig h dos e male s at PND92 . Absolut e an d relativ e increase s i n teste s weight s wer e observe d i n 50 0 pp m male s at PND2 2 an d 5 0 an d relativ e weight s onl ywer e increase d i n 180 0 pp m male s at PND92 . Relativ e prostrat e weigh t wa s increase d i n 10 0 an d 50 0 pp m group s o n day92 ; absolut e epididyma l an d semina l vesicl e weight s wer e decrease d fo r180 0 pp m male s at PND92 . Pituitary , thyroid , testis , epididymi s an d prostat e histolog yresult s showe d n o alteration s acros s th e treatmen t groups ; live rhistolog y showe d hig h dos e male s wit h stron g panlobula rhypertroph yat PND9 2 (4/ 4 animals) , whic h i s consisten t wit h finding s i n th e secon d multigeneratio n reproductio n study . I n tw o multi-generatio n rat studie s (Lose ran d Lorke , 1979 : Eiben , 1984) , P adult s wer e expose d t o triadimefo n i n th e die t fo rat leas t 1 0 week s prio rt o matin g an d the n throughou t mating , gestation , an d lactatio n periods . Indirec t exposur e fo rth e F 1 offsprin g covers th e i n uter o an d lactatio n period s wit h direct exposur e startin g aroun d PN D 1 1 o r1 2 whe n pup s begi n t o consum e treate d die t an d fro m PN D 28 , continue s fo rat leas t 1 0 week s prematin g throughou t mating , gestation , an d lactatio n period s fo rpup s selecte d t o becom e F 1 parents . Thus , th e exposur e duratio n fo rth e F 1 generatio n exceed s that require d b yth e puberta l assa yan d therefor e provide s a more sever e assessmen t o fa compound' s potentia l t o affec t endocrine relate d endpoints . Th e result s o fthes e studie s hav e bee n discusse d above . Ther e wer e n o pattern s o ffertilityo rse xorga n relate d finding s i n eithe rmale s o rfemale s tha t sugges t androge n disruption . Th e puberta l assa yi s designe d t o detect effect s o n puberta l developmen t an d thyroi d function . A s detaile d i n Tabl e 3 , al l o fth e sexua l developmenta l endpoint s include d i n a mal e puberta l assa yhav e bee n assesse d fo rtriadimefon . Th e result s o fthes e studie s have bee n discusse d above . There wa s n o obviou s patter n o ffertilityo rse xorga n relate d finding s i n eithe rmale s o r female s that indicat e disruptio n o fth e androge n system . Wit h th e exceptio n o fweight s o faccessorytissue s an d clinica l chemistry , th e endpoint s i n th e mal e puberta l assa yare capture d i n th e two-generatio n rat reproductio n studie s an d th e 200 7 publishe d researc h b yGoet z e t al. . I n addition , complet e sper m analyse s wa s performe d i n bot h regulator yan d researc h studie s detaile d abov e a s wel l a s i n a stud ypublishe d b yTully , e t al . i n 200 6 i n whic h adul t mal e S D rat s ( 6 pe rdos e group ) receive d triadimefo n b ygavag e fro m PN D 6 0 t o 7 4 at dos e level s o f5 , 5 0 o r11 5 mg/kg bw/da ywithou t advers e impac t o n sper m morpholog yo rmotility , testi s o repididymi s weight s o rtesti s o rthyroi d histopathology . Complet e clinica l chemistr ydat a are als o availabl e fo rmultipl e timepoint s i n th e chroni crat an d do g studies , a s i s additiona l thyroi d weigh t an d histolog yfro m chroni cstudie s i n rat s an d dogs . Th e result s fro m al l o fthes e studies , i n particula rth e two-generatio n rat reproductio n studies , developmenta l neurotoxicitystud y(Sheet s e t al. , 2008 ) an d 200 7 publicatio n b yGoet z e t al . shoul d b e considere d adequat e t o addres s th e dat a requiremen t fo ra mal e puberta l assay . Therefore , th e nee d t o conduct thi s assa yshoul d b e waived . I n viv o Hershberge rAssa y Th e Hershberge rassa yi s a short-ter m i n viv o assa yt o detect androgeni co rantiandrogeni c chemical s o rchemical s tha t inhibi t 5a-reductas e b ymeasurin g weigh t change s i n fiv e androge n dependen t tissues : ventra l prostat e (VP) , semina l vesicl e (SV ) (plu s fluid s an d Pag e 1 4 of4 6

# Baye r

CropScienc e US004 6 coagulatin g glands) , levato rani-bulbocavernosu s (LABC ) muscle , paire d Cowper' s gland s (COW ) an d th e glan s peni s (GP) . Sufficien t dat a o n thes e specifi ctissue s are no t availabl e i n guidelin e o ri n researc h studie s reporte d i n th e publi cdomain , an d therefore th e Consortiu m wil l conduct th e Hershberge rassay . Conclusio n fo rA R Assay s Advers e effect s tha t woul d b e expecte d fro m exposur e t o a n androge n disruptin g chemica l ar e show n below . Th e effect s are no t consisten t wit h what ha s bee n observe d i n mammalia n toxicolog ystudie s wit h triadimefon . Mechanis m Potentia l Effect s i n Male s Potentia l Effect s i n Female s Androgenicity Increase d o rdecrease d mal e reproductiv e orga n weight s Reduce d sper m count s Testicula ratroph y Increase d anogenita l distanc e Accelerate d vagina l openin g Reduce d fertility Altere d differentia l follicl e coun t Histopathologica l finding s o fth e femal e reproductiv e organ s (e.g. , vagina l agenesis ) Induce d mal e se xaccessor y tissue s Anti-androgenicity Delaye d preputia l separatio n Reduce d anogenita l distanc e Ectopi cteste s Hypospadias/epispadias Reduce d fertility Reduce d reproductiv e orga n weigh t (i.e. , prostate , semina l vesicles ) Retaine d nipples/areola s Histopathologica l finding s o fth e reproductiv e organ s (e.g. , epididyma l agenesis , testicula rtumors ) •Altere d pu p se xratio s betwee n externa l an d interna l sexin g

I n summary , scientifi cinformatio n indicate s that a n androgeni ccompoun d woul d produc e amon g othe rfinding s change s i n sexua l development , estrou s cyclicity , fertilityan d othe r reproductiv e endpoints , a s wel l a s t o produc e weigh t an d histopathologica l change s t o reproductiv e an d sexua l accessoryorgans . Adequat e informatio n o n triadimefo n effect s o n mos t parameters i s availabl e i n existin g regulator ystudie s an d publications . Give n th e leve l o f availabl e information , i t i s considere d unnecessar yt o conduct eithe rth e androge n recepto r bindin g assa yo rt o sacrific e th e additiona l animal s neede d t o conduct th e mal e puberta l i n viv o androge n screenin g assays . I n orde rt o addres s uncertaintie s regardin g orga n weigh t effect s an d increase d testosteron e level s reporte d i n th e Goet z publication , a Hershberge rassa ywil l b e conducted . Thyroi d Syste m OSR I Assay s (rat pubertal s an d amphibia n metamorphosis ) hav e bee n include d i n th e EDS P Tie r1 screenin g batter

yt o asses s th e potentia l o fa chemica l t o interfer e wit h thyroi d function . Althoug h thes e assay s a s specificall ydescribe d i n EPA' s OPPT S 89 0 Guidelin e Serie s hav e no t Pag e 1 5 of4 6

## Bayer

CropScienc e US0046 bee n conducte d wit h triadimefon , adequat e informatio n exist s t o asses s triadimefo n potentia l t o interfere wit h th e hypothalamic-pituitary-thyroida l (HPT) axis . A s describe d previously , informatio n o n thyroi d weigh t an d histolog yare availabl e fro m guidelin e studie s includin g lifetim e feedin g studie s i n rat s an d mice , an d a chroni cdo g stud ya s wel l a s a numbe ro f publication s b yEP Aresearchers . Rockett e t al . (2006 ) an d Goet z e t al . (2007 ) evaluate d triadimefo n effect s o n thyroi d weigh t an d histopatholog yi n developin g femal e an d mal e rats , respectively , expose d i n utero , throughou t lactation , an d u p t o PN D 9 2 fo rmale s an d PN D 9 8 fo rfemales . I n addition , thyroi d hormon e leve l measurement s at variou s postnata l age s hav e bee n conducte d b yEP Aresearcher s (Goet z e t al. , 2007 , Tull yet . al , 200 6 bot h previousl y discusse d an d Wol fe t al. , 200 6 discusse d below) . N o histopathologica l change s wer e observe d i n do g o rmous e thyroi d (o rpituitary) gland s i n th e lon g ter m regulator

ystudie s discusse d earlie r(Bombard , 1986 , Hoffma n an d Groning , 1978) . I n th e rat lifetim e feedin g stud y(Bomhard an d Schilde , 1991) , histopathologi cchange s wer e observe d i n th e thyroi d i n rat s administere d triadimefo n i n th e die t at 180 0 ppm:114/19 9 (male/female ) mg/k g bw/da y(HOT) , bu t onl yi n animal s sacrifice d at stud ytermination , i.e . afte r approximatel y2 year s o fexposure ; n o effect s o n th e thyroi d wer e observe d i n animal s tha t wer e sacrifice d afte r1 year . Th e change s consiste d o fa n increase d incidenc e o fcysti chyperplasi a an d a n increase d incidence o fthyroi d adenoma s i n bot h sexes , whic h wer e suggeste d b yBC S t o b e withi n historica l contro l rang e (Sander , E . an d Schilde , B. , 1993) . EPA' s Cance rPee r Revie wCommitte e conclude d that th e carcinogeni cpotentia l o ftriadimefo n i s equivoca l base d o n thes e findings . I n th e 200 6 stud yb yEP Aresearchers Wol fe t al. , th e triadimefo n mod e o factio n fo rthyroi d tumorigenesi s wa s investigated . Seve n wee k ol d Wista rHa n mal e rat s receive d triadimefo n i n th e die t at concentration s o f100 , 50 0 o r180 0 pp m fo r4 , 3 0 o r9 0 day s (1 3 animal s pe rdos e group) . Thyroi d gland s (amon g others ) wer e take n at necrops yan d score d fo rcolloi d depletion , follicula rcel l hypertroph yand/o rfollicula rcel l hyperplasia . Tota l circulatin g seru m thyroi d hormon e level s wer e analyze d a s wel l a s cholesterol , triglyceride s an d hig h densit ylipoproteins . Hepati cenzym e activitywa s assessed . Th e histolog yo fth e thyroi d wa s onl ydifferen t fro m contro l i n rat s treate d wit h th e hig h dos e o ftriadimefo n fo r3 0 days . Thes e rat s ha d colloi d depletio n an d follicula rcel l hypertrophy , whic h wer e no t differen t fro m contro l fo rrat s treate d fo r 9 0 days . Fro m hormon e leve l an d enzym e activityfinding s th e authors conclude d tha t althoug h triadimefo n induce s UDPG Tactivityan d reduce s seru m T4 levels , ther e i s n o associate d increas e i n TS H level s o rpersistentl yincrease d thyroi d follicula rcel l proliferation . However , BC S suggest s tha t th e fact tha t a compensator yTS H increas e wa s no t see n mayhave bee n becaus e th e impac

t o n th e thyroi d ove rth e duratio n ofdosin g wa s not severe enoug h o rth e timin g o fbloo d samplin g wa s no t optimal . Base d upo n th e findin g o fincrease d UDPG Tactivity , th e alteratio n o fthyroi d hormon e balanc e secondar yt o live rinductio n remain s a plausibl e mechanism . Othe rdownstrea m effect s whic h migh t b e indicativ e o finterferenc e o fth e thyroi d syste m includ e change s i n endpoint s suc h a s acousti cstartl e an d moto ractivit yi n th e DN Tstud yan d effect s o n fetal/pu p weigh t an d survival . N o effect s o n an yo fthes e parameters wer e observe d i n th e offsprin g o fdam s administere d die t containin g triadimefo n at level s o f10 0 an d 30 0 pp m fro m gestatio n day6 throug h lactatio n day2 1 (Sheet s e t al. , 2008) . At 80 0 ppm , pre-weanin g bod y weigh t an d bod yweigh t gai n wer e reduce d fo rhig h dos e pup s fo rbot h sexe s an d ther e wa s a n increase d pea k

amplitud e durin g auditor ystartl e refle xtestin g i n hig h dos e female s considere d Pag e 1 6 of4 6m\



# H NBayer

CropScienc e US004 6 equivocal . I n developmenta l toxicit ystudie s i n rat s (Unger , 1982 , Nagum o e t al. , 1981 ) an d rabbit s (Clemen s an d Hartnagel , 1990) , ther e wer e n o treatment-relate d effect o n gestatio n o r fertilityindice s a t th e highes t dose s tested . Ra t feta l weight s wer e unaffecte d b ytreatment ; rabbi t feta l weight s wer e slightl yreduce d i n hig h dos e pups . Advers e effect s tha t woul d b e expecte d fro m exposur e t o a thyroi d functio n disruptin g chemica l are show n below . Mechanis m Potentia l Effect s i n Male s Potentia l Effect s i n Female s Thyroi d hormon e modulatio n Decrease d T 3 and/o rT 4 levels , increase d TS H level s Increase d thyroi d weight s Decrease d colloi d conten t an d thyroi d follicl e siz e Follicula rcel l hypertroph y Thyroi d follicula rcel l hyperplasi a Thyroi d follicula rcel l tumors Th e Amphibia n Metamorphosi s Assa y(AMA ) involve s th e us e o ftadpole s t o determin e i f chemical s affec t th e hypothalamic-pituitary-thyroi d (HPT ) axi s durin g metamorphosi s an d consequentl yresult s i n developmenta l effect s (USEPA , 2009a) . Th e AM Arepresent s a generalize d vertebrat e mode l t o th e exten t tha t i t i s base d o n th e conserve d structur e an d functio n o fth e HP Taxi s (USEP A2009b) . Arevie wo fexistin g thyroi d assay s wa s reporte d b y OEC D i n a detaile d revie wpape r(OEC D 2006) . Thi s repor t point s ou t th e conserve d natur e o f th e HP Taxis . A s outline d above , a numbe ro fstudie s hav e bee n conducte d t o investigat e th e potentia l effect s o ftriadimefo n o n thyroi d system s o fvertebrates . Thes e studie s wer e als o don e wit h conserve d model s s o a n additiona l evaluatio n wit h a n additiona l species , wit h th e highl y conserve d HP Taxis , i s unnecessar yt o evaluat e th e thyroi d activit ypotentia l i n vertebrates . Th e substantia l existin g dat a fro m studie s shoul d b e deeme d adequat e fo ra screenin g leve l thyroi d evaluation . Therefore , th e conduct o fa n AM Ai s unnecessar yan d woul d resul t i n th e expenditur e o fresource s an d unnecessar ysacrific e o fanimals . Result s o fguidelin e an d publishe d studie s indicat e tha t repeate d exposur e t o triadimefo n at sufficien t dose s doe s resul t i n microscopi cchange s i n th e rat thyroid . Adequat e informatio n o n triadimefo n effect s o n al l parameters i s availabl e i n existin g regulator ystudie s an d publications . Give n th e leve l o favailabl e information , i t i s considere d unnecessar yt o conduct eithe ro fth e rat puberta l o rth e amphibia n metamorphosi s assays . Therefore , th e requiremen t fo rth e thyroi d functio n Tie r1 assay s shoul d b e waived . Steroi d Synthesi s OSR I Triadimefo n wa s include d i n EPA' s aromatas e assa yvalidatio n stud yan d result s are reporte d i n EPA' s Decembe r11 , 200 7 "Integrate d Summar yRepor t o n Aromatase" " an d fulfil l th e requiremen t fo rth e aromatas e assay . Th e endpoint s discusse d abov e fo rth e estroge n an d androge n system s ar e als o relevan t fo r assessmen t o fa compound' s potentia l t o interfer e wit h steroidogensis . Thes e endpoint s ar e show n i n Tabl e 1 a an d thos e considere d a s functionall yequivalen t t o addres s th e purpos e o f th e steroidogenesi s assay s are indicated . I n addition , a stud yb yEP Aresearcher s (Goetz , e t al. , 2009 ) report s result s ofi n vitr o assay s i n rat testi s an d H295 R cell s a s wel l a s i n viv o seru m an d testi s testosteron e level s i n mal e rat s afte r1 4 o r3 0 day s dietar yexposure . Pag e 1 7 o

f4 6

# Bayer

CropScienc e US004 6 Regardin g th e i n vitr

o assay , onl y5 rathe rtha n 7 concentration s wer e teste d i n th e H295 R cel l assay , an d onl y1 contro l wa s use d instea d o fth e 2 specifie d fo rOPPT S 890.1550 . I n additio n t o testosteron e an d estradiol , progesteron e level s wer e als o measured . Th e autho rconclude d that triadimefo n i s a wea k inhibito roftestosteron e productio n i n vitro . Thi s stand s i n contras t t o th e i n viv o finding s wer e testosteron e level s wer e elevate d (discusse d below) . Th e publishe d stud yi s considere d adequat e t o fulfil l th e requiremen t fo rth e i n vitr o steroidogenesi s assay . Regardin g th e i n viv

o investigation , mal e Wista rha n rat s (PN D 60 ) wer e administere d triadimefo n i n th e die t at 180 0 pp m fo r3 0 days . Bod yweigh t an d foo d consumptio n wer e measure d weekly ; animal s were tai l ble d o n da y1 4 fo rtestosteron e measurements . O n da y3 0 animal s wer e decapitate d an d trun k bloo d collecte d fo rseru m measurement s fo rtestosterone , estradiol , L H an d prolactin . Teste s wer e weighe d an d sna p frozen ; liver , epididymis , ventra l prostate , semina l vesicle s an d pituitar ywer e collecte d an d weighed . Bod yweight s wer e lowe ri n treate d rat s relativ e t o contro l throughou t th e cours e o fth e study . Seru m testosteron e level s wer e no t increase d afte r1 4 days bu t wer e significantl yincrease d at 3 0 day s (ca . 290 % change) , a s wa s intra-testicula rtestosteron e (210 % change) . Seru m level s o festradiol , L H an d prolacti n were somewha

t elevate d a

t 3 0 days , bu t increase s were not statisticall ysignificant . Live r weights , bot h absolut e an d relative , wer e significantl yincrease d afte r3 0 day s o fexposur e (2 7 an d 37% , respectively) . Absolut e pituitar yan d paire d epididyma l weights , bu t no t relative , wer e reporte d t o b e somewha t decrease d (9. 8 an d 5.2% , respectively , p<0.05) . Androge n dependen t ventra l prostat e an d semina l vesicl e weight s wer e unaffected , whic h th e autho rstate s ar e consisten t wit h othe rstudie s whic h d o no t sho wmuc h evidenc e o fA R antagonis m i n androgen dependen t tissue s (citin g Goetz , e t al. , 2007 ; Tull ye t al. , 200 6 ; Okub o e t al. , 2004) . Goet z e t al . conclud e tha t triadimefo n appears t o inhibi t CYP17A 1 i n vitro , decrease s testosteron e level s i n vitro , bu t increase s rat seru m an d testi s testosteron e level s i n viv o afte r sufficien t duratio n o fexposur e b ya n unresolve d mechanism . Advers e effect s that woul d b e expecte d fro m exposur e t o a chemica l inhibitin g steroidgenesi s o r aromatas e ar e show n below . Mechanis m Potentia l Effect s i n Male s Potentia l Effect s i n Female s Reduce d steroi d biosynthesi s Simila rt o anti-androgenicity Possibl e increase d seru m cholestero l level s Increase d Leydi g cel l tumor s Simila rt o anti-estrogenicity / aromatas e inhibitio n Aromatas e inhibitio n Increase d tim e t o matin g Decrease d mal e mountin g behavio r Decrease d bod yweigh t (chronic ) Increase d testi s weigh t (chronic ) Increase d bod yweigh t Decrease d uterin e weigh t Increase d ovarysiz e Polycysti covarie s Stroma l hyperplasi a i n ovary (chronic ) Hyalinizatio n i n ovar

y(chronic ) Increase d urete ran d bladde r infectio n Decrease d mammarytumo r incidenc e (i n Spragu e Dawle yrats ) Pag e 1 8 of4 6A I B AY E F 

# Baye r

CropScienc e US004 6 Give n th e leve l o favailabl e informatio n fro m U S EP Apublishe d research , th e requiremen t fo r aromatas e an d steroidgenesi s assay s i s fulfilled . Overal l Conclusio n Th e Triadimefo n Consortiu m believe s that existin g dat a o n triadimefo n are fo rth e mos t par t adequat e t o asses s th e potentia l fo rthi s compoun d t o interac t wit h estrogen , androge n an d thyroi d systems . However , i n orde rt o addres s equivoca l finding s an d suppl yadditiona l studie s tha t wil l provid e complementar ydat a an d sufficien t scientifi credundanc yt o assur e tha t sufficien t definitiv e dat a ar e availabl e t o perfor m a weigh t o fevidenc e (WOE ) evaluatio n o f whethe ran yTie r2 testin g woul d b e necessary , th e Triadimefo n Consortiu m wil l conduc t a Fis h Shor t ter m Reproductio n Assa yan d a Hershberge rBioassay . Th e Triadimefo n Consortiu m request s waivers fro m al l othe rTie r1 studie s a s justifie d below . Endocrin e Disrupto rScreenin g Progra m Tie r1

Tes t Justificatio n I n Vitr o Screenin g Assay s OPPTS 890.1250 : Estroge n Recepto rBindin g Assa y Usin g Rat Uterin e Cytoso l (ER-RUC ) OPPT S 890 . 1300 : Estroge n Recepto r Transcriptiona l Activatio n (Huma n Cel l Lin e (HeLa 9903) ) OPPT S 890.1150 : Androge n Recepto rBindin g (Ra t Prostat e Cytosol ) OPPT S 890 . 1200 : Aromatas e (Huma n Recombinant ) OPPT S 890 . 1550 : Steroidogenesi s (Huma n Cel l Lin e -H295R ) Functiona l equivalence ofOSR I Functiona l equivalenc e o fOSR I Functiona l equivalenc e o fOSR I Include d a s a tes t substanc e i n EPA' s validatio n o fth e assa y Functiona l equivalenc e o fOSR I plu s publicatio n assesse d a s sufficientl yequivalen t t o protoco l I n Viv o Screenin g Assay s OPPT S 890 . 1600 : Uterotrophi cAssa y OPPT S 890.1450 : Puberta l Developmen t an d Thyroi d Functio n i n Intac t Juvenile/

Peripuberta l Femal e Rat s OPPT S 890.1500 : Puberta l Developmen t an d Thyroi d Functio n i n Intac t Juvenile/

Peripuberta l Mal e Rat s OPPT S 890.1100 : Amphibia n Metamorphosi s (Frog ) Functiona l equivalenc e o fOSR I Functiona l equivalenc e o fOSR I Functiona l equivalenc e ofOSR I Functiona l equivalenc e o fOSR I

Pag e 1 9 of4 6

# Baye r

CropScienc e US004 6 REFERENCE S Th e followin g studie s hav e bee n submitte d previousl yt o EP Aan d are cite d b yMRI D number : MRI D No . 3253 9 Hoffman , K . an d Groning , P . (1978 ) Me b 644 7 Long-Ter m ToxicityStud yo n Dog s (Two-Yea rFeedin g Study) : Repor t No . 7882 ; Repor

t No . 66656 . (Unpublishe d stud y receive d Ap r28 , 198 0 unde r3125-318 ; prepare d b yBayer , AG , submitte d b yMobayChemica l Corp. , Kansa s City , Mo. ; CDL099413-D) . MRI D No . 3254 1 Loser , E . an d Lorke , D . (1979 ) Me b 644 7 Multigeneratio n Reproductio n Stud yo n Rats : Repor

t No . 8297 ; Repor

t No . 67752 . (Unpublishe d stud yreceive d Ap r28 , 198 0 unde r3125-318 ; prepare d b yBayer , AG , submitte d b yMobayChemica l Corp. , Kansa s City , Mo.;CDL : 099413-F ) MRI D No . 4862 4 Mohr , U . (1976 ) Me b 6447 : Subchroni cToxicityStud yo n Rat s (Twelv e - Wee k Feedin g Experiment) : Repor t No . R 84 0 a ; Repor

t No . 44300 . (Unpublishe d stud y receive d Au g 15 , 1977 unde r3125-318 ; prepare d b yMedizinisch e Hochschul e Hannover , Abteilun g fu rExperi - mentell e Pathologie , Wes t Germany , submitte d b yMobayChemica l Corp. , Kansa s City , Mo. ; CDL231312-C ) MRI D No . 4862 5 Hoffman , K.an d Luckhaus , G . (1974 ) Me b 6447 : Subchroni cToxicityStud y o n Dog s (Thirteen-Wee k Feedin g Experiment) : Repor t No . 5071 ; Repor

t No . 44737 . (Unpublishe d stud yreceive d Au g 15 , 1977 unde r3125-318 ; prepare d b yBayer , AG , submitte d b yMobayChemica l Corp. , Kansa s City , Mo. ; CDL231312-D) . MRI D No . 6022 6 Hoffmann , K. , Luckhaus , G . an d Lorke , D . (1974 ) Me b 6447 : Subchroni c ToxicityStud yo n Dog s (Thirteen-Wee k Feedin g Experiment) : Repor t No.5071 ; Repor

t No . 44737 . (Unpublishe d stud yreceive d De c15 , 197 7 unde r3125-EX-149 ; prepare d b yBayer , AG , W . Germany , submitte d b yMobayChemica l Corp. , Kansa s City , Mo. ; CDL : 232490-D) . MRI D No . 8902 3 Nagumo , K. , Teraki , Y . an d Chiba , T . (1981 ) Teratogenicity Tes t o fME B 644 7 i n Pregnan t Rats : Submitte r80257 . (Unpublishe d stud yreceive d Ja n 5 , 198 2 unde r1F2474 ; prepare d b ySt . Mariann a Univ. , First Dept . o fAnatomy , Laborator yo f Embryology , submitte d b yMobayChemica l Corp. , Kansa s City , Mo. ; CDL : 070570-A ) MRI D No . 12626 1 Hoffmann , K. , Groning , P . an d Lorke , D . (1978 ) ME B 6447 : Long-ter m Toxicit yStud yo n Dog s (Two-yea rFeedin g Study) : Repor t No . 7882 ; 66656 . (Unpublishe d stud y receive d Ma r11 , 198 3 unde r3125-346 ; prepare d b yBaye rAG , W . Ger. , submitte d b yMobay Chemica l Corp. , Kansa s City , MO ; CDL:071468-B ) MRI D No . 14933 6 Unger , T. , Va n Goethem , D . an d Shellenberger , T . (1982 ) ATerato logica l Evaluatio n o fBayleto n i n Mate d Femal e Rats : MR I Project No . 7272-B . Unpublishe d stud yprepare d b yMidwes t Researc h Institute . 5 2 p . MRI D No . 15507 5 Eiben , R . (1984 ) ME B 644 7 (Triadimefon) : Two-generatio n Stud ywit h Rats : Supplementar

yStudy : Repor

t No . 12712 . Unpublishe d stud yprepare d b yBaye rAG , Institut e ofToxicology . 16 8 p . Pag e 2 0 o f4 6

# Bayer

CropScienc e US004 6 MRI D No . 4075210 1 Bombard , E . (1986 ) ME B 6447 : CarcinogenicityStud yo n NMR I Mice (21 - Mont h Administratio n i n th e Feed) : Repor t No . 87287 . Unpublishe d stud yprepare d b yBaye r AG . 119 0 p . MRI D No . 4086510 1 Bombard , E . an d Hahnemann , S . (1986 ) ME B 6447 : Commo n Name : Triadimefon , th e Activ e Ingredien t o fBayleton : CarcinogenicityStud yo n NMR I Mice : Project I D 87287 . Unpublishe d stud yprepare d b yBaye rAg . 5 p . MRI D No . 4144620 1 Clemens , G . an d Hartnagel , R . (1990 ) Teratolog yStud yi n th e Rabbi t wit h ME B 644 7 (Triadimefon) : Repor t No. : MTD0149 . Unpublishe d stud yprepare d byMiles , Inc . 15 5 p . MRI D No . 4208960 1 Clemens , G. ; Hartnagel , R . (1991 ) Teratolog yStud yi n th e Rabbi t wit h ME B 6447 : Supplemen t t o MRI D 41446201 : La b Project Number : 100026 : MTD0149 . Unpublishe d stud yprepare d byMiles , Inc. , Tox . Dept . 1 1 p . MRI D No . 4215390 1 Bomhard , E. ; Schilde , B . (1991 ) ME B 6447 : Chroni cToxicit yan d CancerogenicityStudie s o n Wista rRat s wit h Administratio n i n Diet ove ra Perio d o f10 5 Weeks : La b Project Number : 20774 : 101922 . Unpublishe d stud yprepare d b yBaye rAg. , Dept . o f Toxicology . 102 9 p . Als o se e MRI D No . 4308930 1 MRI D No . 4308930 1 Sander , E . an d Schilde , B . (1993 ) Supplement : Th e Incidenc e of Thyroi d Follicula rCel l Tumor s i n Wista rRats : (BAYLETO N ME B 6447) : ACompilatio n o f Historica l Data : La b Project Number : T/3027626 : 20774/A . Unpublishe d stud yprepare d b y Baye rAG . 5 7 p . MRI D No . 4737710 1 Sheets , L. ; Gilmore , R. ; Hoss , H . (2008 ) ADevelopmenta l Neurotoxicity Stud ywit h Technica l Grad e Triadimefo n i n Wista rRats . Project Number : 07/D72/IL , 201766 . Unpublishe d stud yprepare d byBaye rCropScience . 1179 p . MRI D No . 9218801 8 Eigenberg , D . (1990 ) MobayCorporatio n Phas e 3 Summar yo fMRI D 00149336 . ATeratologica l Evaluatio n o fBayleto n (Triadimefon)i n Mate d Femal e Rats : Project 7272- B (MobayRepor

t 82270) . Prepare d b yMIDWES TRESEARC H INSTITUTE . 2 2 p . MRI D No . 9218801 9 Eigenberg , D . (1990 ) MobayCorporatio n Phas e 3 Summar yo fMRI D 0015507 5 an d Relate d MRID s 00032541 . ME B 644 7 (Triadimefon ) Multigeneratio n Reproductio n Stud yo n Rats ; (Triadimefon ) Two-generatio n Stud ywit h Rat s (Bayleton) : Project s 829 7 an d 1271 2 (MobayReport s 6775 2 an d 82270) . Prepare d b yBAYE R AG . 3 8 p . MRI D No . 9218802 0 Loser , E . (1990 ) MobayCorporatio n Phas e 3 Reforma t o fMRI D 0003254 1 an d Relate d MRID s 00155075 . ME B 644 7 (Triadimefon ) Multigeneratio n Reproductio n Stud yo n Rats : Project Numbe r67752 ; Repor t No . 8297 . Prepare d b yBAYE R AG . 17 3 p . Th e followin g guideline s an d revie wpape rwer e use d fo rreference : OECD , 2006 . Draft Detaile d Revie wPape ro n Thyroi d Disruptio n Assays . OEC D Environment , Healt h an d SafetyPublicatio n Serie s o n Testin g an d Assessment . No . no t assigned . Paris , France . Pag e 2 1 o f4 6

# Baye r

CropScienc e US004 6 USEPA , 2009a . Tes t Order/DC I # : EDSP-109901-15 6 fo rTriadimefon . Dat e Issued : 12/17/2009 . USEPA , 2009b . Offic e o fPrevention , Pesticide s an d Toxi cSubstances . Tes t Guideline , 890.1100 : Amphibia n Metamorphosi s (Frog) . EP A740-C-09-002 . Octobe r2009 . U.S . Environmenta l Protectio n Agency . Washington , D.C . Othe rPublishe d literature : Goetz , A. , Ren , H. , Schmid , J. , Blystone , C. , Thillainadarajah , I. , Best , D. , Nichols , H. , Strader , L , Wolf , D. , Narotsky , M. , Rockett , J . an d Dix , D . (2006) . Toxicol . Sci . 95(1) : 227-239 . Goetz , A. , Rockett , J. , Ren , H. , Thillainadarajah , I . an d Dix , D . (2009) . Syst . Biol . Reproof . Med . 55 : 214-226 . Klimisch , H.-J. , Andreae , M an d Tillman , U . (1997) . ASystemati cApproac h fo rEvaluatin g th e QualityofExperimenta l Toxicologica l an d Ecotoxicologica l Data . Regulator yTox . an d Pharm . 25 : 1-5 . Kojima , H. , Katsura , E. , Takeuchi , S. , Niiyama , K. , an d Kobayashi , K . (2004) . Screenin g fo r estroge n an d androge n recepto ractivitie s i n 20 0 pesticide s byi n vitr o reporte rgen e assay s usin g Chines e Hamste rOvar ycells . Environ . Health Perspect . 112 : 524-531 . Okubo , T. , Yokoyamo , Y. , Kano , K. , Soya , Y . an d Kano , I . (2004) . Arch . Environ . Contam . Toxicol . 46 : 445-453 . Petit , F. , L e Goff , P. , Crevadi , J-P. , Valotaire , T. , an d Pakdel , F . (1997) . Two complementar y bioassay s fo rscreenin g th e estrogeni cpotenc yo fxenobiotics : recombinan t yeas t fo rtrou t estroge n receptors an d trou t hepatocyt e cultures . J . Mol . Endocrin . 19 : 321-335 . Rockett , J. , Narotsky , M. , Thompson , K. , Thillainadarajah , I. , Blystone , C. , Goetz , A. , Ren , H. , Best , D. , Murrell , R. , Nichols , H. , Schmid , J. , Wolf , D . an d Dix , D . (2006) . Effect ofconazol e fungicide s o n reproductiv e developmen t i n th e femal e rat . Reprod . Toxicol . 22 : 647-658 . Tully , B. , Bao , W. , Goetz , A. , Blystone , C. , Ren , H. , Schmid , J. , Strader , L. , Wood , C. , Best , D. , Narotsky , M. , Wolf , D. , Rockett , J . an d Dix , D . (2006) . Gen e expressio n profilin g i n live ran d testi s ofrat s t o characteriz e th e toxicityoftriazol e fungicides . Toxicol . Appl . Pharmacol . 215 : 260-273 . Vinggaard , A.M. , Breinholt , V . an d Larsen , J.C . (1999) . Screenin g o fselecte d pesticide s fo r oestroge n recepto ractivatio n i n vitro . FoodAddit . Contam . 16(12) : 533-542 . Wolf , D. , Allen , J. , George , M. , Hester , S. , Sun , G. , Moore , T. , Thai , S-H. , Delker , D. , Winkfield , E. , Leavitt , S. , Nelson , G. , Roop , B. , Jones , C. , Thibodeaux , J . an d Nesnow , S . (2006) . Toxicit y profile s i n rat s treate d wit h tumorigeni can d non-tumorigeni ctriazol e conazol e fungicides : propiconazole , triadimefo n an d myclobutanil . Toxicologic . Pathology34 : 895-902 . Pag e 2 2 o f4 6

# Bayer

CropScienc e US004 6 Tabl e 1a : Functiona l Equivalenc e Matri x- Apica l Endpoint s i n Existin g Tridimefo n Studie s Relevan t t o Evaluatio n o fPotentia l Effect s Resultin g fro m Mode s o fActio n Assesse d i n th e I n Vitr o Assay s

OPPT S 870.380 0 Two-Generatio n Rat Reproductio n (combine d MRID s 0032541 , 155075 ) •Reproductiv e indice s (F O an d F 1 parents ) •Litte rparameters (F 1 an d F2 J •Pu p Se xRati o •Externa l genitali a anomalie s (F 1 an d F2 } •Orga n wt s ovarie s (F O an d F 1 parenta l 9 ) •Orga n histopathology : uterus , ovaries , vagin a (F 2 pup s only ) (F 1 parental , F 2 an d F 3 pup s 9 ) •Orga n wts : epididymide s + teste s (F O an d F 1 parenta l CT) •Orga n histopathology : semina l vesicle s an d prostat e (F 1 parenta l only) , epididymides , teste s (F 1 parental , F 2 an d F 3 pup s C? ) •Orga n histopathology : pituitar y(F 1 parenta l an d F 2 pups ) adrenal s (F 1 parental , F 2 an d F 3 pups ) an d thyroi d (F 3 pups ) •Ovaryfollicula rstatu s a t necrops y (F 1 parental , F2 an d F 3 pup s § ) •Andrologica l parameters (sper m count , motility , productio n an d anomalie s - F O an d F 1 parenta l C? ) OPPT S 870.630 0 Developmenta l neurotoxicity(MRI D 47377 1 01 ) •Pu p Se xRati o •Vagina l Openin g •Preputia l Separatio n •Externa l genitali a anomalie s OPPT S 870.310 0 Subchroni ctoxicit yi n rat s (MRI D 48624 ) •Orga n wt s an d histopathology : ovarie s •Orga n wt s an d histopathology : E R bindin g an d transcriptiona l activatio n

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teste s •Orga n wt s an d histopathology : adrenal , pituitary , thyroi d OPPT S 870.315 0 Subchroni ctoxicit yi n dog s (MRI D 48625 , 60226 ) •Orga n wt s an d histopathology : ovaries , uteru s (histopat h only ) •Orga n wt s an d histopathology : prostate , testes , epididymide s (histopat h only

) •Orga n wt s an d histopathology : adrenal , pituitary , thyroi d OPPT S 870.4 1 OO b Chroni ctoxicit yi n dog s (MRID s 32539 , 126261 ) •Orga n wt s an d histopathology : ovaries ; histopat h onl ymammar y glands , uteru s •Orga n wt s an d histopathology : prostate , testes ; histopat h onl y semina l vesicles , epididymide s •Orga n wt s an d histopathology : adrenal , pituitary , thyroi d OPPT S 870.420 0 Mous e carcino genicity(MRI D 40752101 , 40865101 ) •Orga n histopathology : mammar y glands , uterus , ovaries , vagin a •Orga n histopathology : semina l vesicles , epididymides , prostates , teste s •Orga n histopathology : adrenal , pituitary , thyroi d OPPT S 870.430 0 Rat combine d toxic ity/carcinogenicity(MRI D 4215390 1 ) •Orga n histopathology : mammar y glands , uterus , ovaries , vagin a •Orga n wt s an d histopathology : testes ; histopat h onl ysemina l vesicles , epididymides , prostat e •Orga n wt s an d histopathology : adrenal, ; histopat h onl ypituitary , thyroi d E R bindin g an d transcriptiona l activatio n

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CropScienc e US004 6 Tabl e 1b : Functiona l Equivalenc e Matri x- Apica l Endpoint s i n Existin g Triadimefo n Studie s Relevan t t o Parameters Assesse d i n th e I n Vivo Assay s OPPT S 870.380 0 Two-Generatio n Ra t Reproductio n (combine d MRID s 0032541 , 155075 ) •Growt h (bod yweight ) •Reproductiv e indice s (F O an d F 1 parents ) •Pu p Se xRati o •Externa l genitali a anomalie s (F 1 an d F2 ) •Orga n wts : ovarie s (F O an d F 1 parenta l 9 ) •Orga n histopathology : uterus , ovaries , vagin a (F 2 only ) (F 1 parenta l an d F2an d FSpup s 9 ) •Orga n wts : semina l vesicle s epididymide s + teste s ( F O an d F 1 parenta l cf ) •Orga n histopathology : semina l vesicle s (wit h coagulatin g gland s an d thei rfluids) , epididymide s (wit h contents) , prostate , teste s (F 1 parenta l an d F2 an d F 3 pups cf) •Orga n wts : adrena l an d pituitar y(F O an d F 1 parents ) •Orga n histopathology : adrena l an d pituitar y(F 1 parent s an d F2pups ) •Andrologica l parameter s (sper m count , motility , productio n an d anomalie s - F O an d F 1 parenta l C? ) OPPT S 870.630 0 Developmenta l neurotoxicit

y(MRI D 47377 1 01 J •Pu p Se xRati o •Vagina l Openin g •Preputia l Separatio n •Externa l genitali a anomalie s OPPT S 870.310 0 Subchroni ctoxicit yi n rat s (MRI D 0048624 ) •Orga n wt s an d histopathology : ovarie s •Orga n wt s an d histopathology : teste s

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CropScienc e US004 6 OPPT S 870.315 0 Subchroni ctoxicit yi n dog s (MRI D 0048625 , 60226 ) •Orga n wt s an d histopathology : ovaries , uteru s (histopat h only) •Orga n wt s an d histopathology : prostate , testes , epididymide s (histopat h only) •Orga n wt s an d histopathology : adrenal , pituitary , thyroi d OPPT S 870.4 1 OO b Chroni ctoxicit yi n dog s (MRI D 32539 , 126261 ) •Orga n wt s an d histopathology : ovaries ; histopat h onl ymammar y glands , uteru s •Orga n wt s an d histopathology : prostate , testes ; histopat h onl y semina l vesicles , epididymide s •Orga n wt s an d histopathology : adrenal , pituitary , thyroi d OPPTS 870.420 0 Mous e carcinogenicit y (MRI D 40752101 , 40865101 ) •Orga n histopathology : mammar y glands , uterus , ovaries , vagin a •Orga n histopathology : semina l vesicles , epididymides , prostates , teste s •Orga n histopathology : adrenal , pituitary , thyroi d OPPT S 870.430 0 Rat combine d toxic ity/carcinogenicity{MRI D 429 1 53901 ) •Orga n histopathology : mammar y glands , uterus , ovaries , vagin a •Orga n wt s an d histopathology : testes ; histopat h onl ysemina l vesicles , epididymides , prostat e •Orga n wt s an d histopathology : adre nal ; histopat h onl ypituitary , thyroi d • Puberta l Femal e S

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CropScienc e US004 6 Tabl e 2 : Femal e puberta l assa yendpoint s compare d t o reporte d endpoint s fo rtriadimefo n fro m multigeneratio n rat reproductio n studie s (MRID s 32541,155075) , a developmenta l neurotoxicity stud y(MRI D 4737101 ) an d relevan t finding s fro m publication s "Effect o fconazol e fungicide s o n reproductiv e toxicityi n th e femal e rat " (Rockett et.al. , Reproductiv e Toxicolog y2 2 (2006 ) 647-658 ) Femal e Puberta l Assa y Multigeneratio n Rat Reproductio n Studie s MRI D 3254 1 MRI D 155075 Rockett et . al . Reproductiv e Toxicolog y 2 2 (2006 ) Exposur e Duratio n PN D 2 2 throug h 4 2 fo rfemale s

Dos e level s (diet) F O animal s - throughou t 1 0 week s (32541 ) o r1 4 week s (155075 ) premating , mating , gestation , an d lactatio n period s F 1 animal s - i n utero (a s fetus ) an d lactatio n period s (a s neonates ) followe d b yexposur e durin g 1 0 (32541 ) o r1 4 (155075 ) week s premating , mating , gestatio n an d lactatio n period s fo rF 1 parent s F 2 animal s - i n uter

o (a s fetus ) an d lactatio n period s (a s neonates ) F 3 animal s a s fo rF 2 (3254 1 ) 0,50,300,180 0 0,50,180 0 Mate d dam s -fro m GD 6 - LD2 1 F 1 female s - i n utero (a s fetus ) an d lactatio n period s a s neonates ) an d fro m PN D 2 2 throug h 9 8

0 , 100 , 500 , 180 0 Endpoint s Include d Dail ybod yweigh t Ag e an d bod yweigh t a t sexua l developmen t (i.e . vagina l opening* ) Orga n weight s • Uteru s (blotted ) • Ovarie s (paired ) • Thyroi d (afte r fixation ) • Live r • Kidney s (paired ) • Pituitary • Adrenal s (paired ) Histolog y • Uteru s • Ovarie s • Thyroi d (colloi d are a an d follicula rcel l height ) • Kidne y Estrou s cyclicit y •Ag e a t firs t vagina l estru s afte rvagina l Weekl ybod yweigh t (al l animals )

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Orga n weight s (FO , F 1 parents ) • Brai n • Ovarie s • Live r • Kidney s

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## Bayer

CropScienc e US004 6 openin g •Lengt h o fcycl e • Percen t o fanimal s cyclin g • Percen t o fanimal s cyclin g regularl y Clinica l (serum ) chemistr y • Standard bloo d panel , includin g creatinin e an d bloo d ure a nitroge n Thyroi d hormone s Dat a no t availabl e fro m reproductio n stud ybu t clinica l chemistr

ydat a are availabl e fro m multipl e timepoint s i n chroni crat an d do g studie s o rabnorma l cycle s durin g week s 1-2 , 5- 6 o r9-1 0 afte rV O Seru m estradio l at at PN D 9 9

Measure d i n mal e cohor t study , se e Tabl e 3 *reporte d i n th e triadimefo n guidelin e developmenta l neurotoxicitystud y

Pag e 2 8 o f4 6

## Bayer

CropScienc e US004 6 Tabl e 3 : Mal e puberta l assa yendpoint s compare d t o reporte d endpoint s fo rtriadimefo n fro m multigeneratio n rat reproductio n studie s (MRID s 32541,155075) , a developmenta l neurotoxicity stud y(MRI D 4737101 ) an d relevan t finding s fro m publicatio n "Disruptio n o ftestosteron e homeostasi s a s a mod e o factio n fo rth e reproductiv e toxicit yo ftriazol e fungicide s i n th e mal e rat " (Goet z et.al. , Toxicologica l Science s 95(1) , 227-23 9 (2007) . Mal e Puberta l Assa y Exposure Duratio n PN D 2 3 throug h 5 3 fo rmale s

Dos e level s (diet

) Multigeneratio n Rat Reproductio n Studie s MRI D 3254 1 MRI D 155075 F O animal s -throughou t 1 0 week s (32541 ) o r1 4 week s (155075 ) premating , mating , gestation , an d lactatio n period s F 1 animal s - i n utero (a s fetus ) an d lactatio n period s (a s neonates ) followe d b yexposur e durin g 1 0 (32541 ) o r1 4 (155075 ) week s premating , mating , gestatio n an d lactatio n period s fo rF 1 parent s F 2 animal s - i n utero (a s fetus ) an d lactatio n period s (a s neonates ) F 3 animal s a s fo rF 2 (3254 1 ) 0 , 50 , 300 , 1800 0,50,180 0 Goet z et . al . Reproductiv e Toxicologica l Science s 95(1 ) (2006 ) Mate d dam s -fro m GD 6 -LD2 1 F 1 male s - i n utero (a s fetus ) an d lactatio n period s (a s neonates ) an d fro m PN D 2 2 throug h 12 0

0,100,500,180 0 Endpoint s Include d Dail ybod yweigh t Ag e an d bod yweigh t a t sexua l develop men t (i.e . preputia l separation* ) Orga n weight s • Semina l vesicl e plu s coagulatin g gland s (wit h an d withou t fluid ) • Ventra l prostat e • Dorsolatera l prostat e • Levato r ani/bulbocavernosu s muscl e comple x • Epididymide s (left an d righ t separately ) • Teste s (left an d righ t separately ) • Thyroi d (afte r fixation ) • Live r • Kidney s (paired ) • Pituitar y • Adrenal s (paired ) Histolog y • Epididymi s (one ) Weekl ybod yweigh t (al l animals )

Histolog yF 3 pup s • Brai n Weekl ybod yweigh t (al l animals ) Orga n weight s (FO , F 1 parents ) • Brai n • Testicle s • Live r • Kidney s

Histolog yF 1 parents , F2 pup s at weanin g Weekl yfro m birt h Anogenita l distanc e an d bod yweigh t at PNDO ; Ag e an d bod yweigh t a t PP S Orga n weights , re l an d abs(PND1,22 , 50 , 92 , 1 male/litter) • Liver " ** * • Testes* * • Ventra l prostate* * • Epididymis* * •Semina l vesicle* * •Brain* * •Pituitar y • Others take n an d weighe d bu t no t reporte d i n publication : hypothalamus , hippocampus , thyroid* *

Histolog y • Liver* * ** * Pag e 2 9 o f4 6

# Baye r

CropScienc e US004 6

• Testi s (one ) • Thyroi d (colloi d are a an d follicula r cel l height ) • Kidne y

Clinica l (serum ) chemistr y • Standard bloo d panel , includin g creatinin e an d bloo d ure a nitroge n Thyroi d hormone s • Teste s • Epididymi s • Live r • Kidney s • Adrenal s • Thyroi d (PND28 ) • Brai n • Teste s • Epididymi s (F 1 only) • Prostat e (F 1 only ) • Pituitar y • Live r • Kidney s • Adrenal s Dat a no t availabl e fro m reproductio n stud ybu t clinica l chemistr

ydat a are availabl e fro m multipl e timepoint s i n chroni crat an d do g studie s

Seru m testosterone , Follicula rstimulatin g hormon e Sper m morpholog y& motility • Testes* * • Ventra l prostat e • Thyroid* * ** * • Epididymi s • Pituitar y

T3 , T4 , TSH** * Seru m testosterone** , Estradiol , Luteinizin g hormone* * at PND5 0 an d 9 2 Sper m morpholog y& motility* * *reporte d i n th e triadimefo n guidelin e developmenta l neurotoxicitystud y * * als o examine d i n Tully , et.al. , i n 200 6 ** * als o examine d i n Wolf , et.al. , 200 6

Pag e 3 0 o f4 6

# p A 

# tE 

# 4 Bayer

CropScienc e US004 6 Tabl e 4 : Functiona l Equivalenc e Matri x- Apica l Endpoint s i n Existin g Studie s an d Publication s Relevan t t o Amphibia n Metamorphosi s

Guidelin e Studie s an d Publication s Multi-Generatio n Rat Reproductio n (MRI D 003254 1 an d 155075 ) •F 1 an d F 2 pu p surviva l an d growt h •4 w k ol d F3 b pup s histopatholog y thyroid , adrenal s (MRI D 0032541 ) Developmenta l toxicity(MRIDs) •Feta l surviva l an d weigh t Developmenta l neurotoxicit y (MRID ) •Pu p surviva l an d growt h •Acousti cStartl e •Moto rActivit y •Surfac e rightin g Goet z et . al . Reproductiv e Toxicologica l Science s 95(1 ) (2006 ) •Pu p surviva l an d growt h (bw ) •Seru m hormon e level s T3 , T4 , TS H •Orga n wt s an d histopatholog y thyroid , pituitar

y Rat subchroni ctoxicity(MRI D •Orga n wt s an d histopathology : adrenal , pituitary , thyroi d Wol fet.al. , Toxicologi cPathology , 3 4 (2006 ) •Mal e rat growt h fro m 7 wks (bw ) •Seru m hormon e level s T3 , T4 , TS H (age s 53 , 79 , 139d ) •Histopatholog ythyroi d (age s a s above ) Do g subchroni ctoxicit y(MRI D •Orga n wt s an d histopathology : adrenal , pituitary , thyroi d Rat combine d toxicity/carcinogen icity(MRID ) •Orga n wt s an d histopathology : adrenal , pituitary , thyroi d Chroni ctoxicit yi n dog s (MRID ) •Orga n wt s an d histopathology : adrenal , pituitary , thyroi d Mous e carcinogenicity(MRID) •Orga n wt s an d histopathology : adrenal , pituitary , thyroi d Amphibia n Metamorphosi s Developmenta l Stage , Lengt h an d Weigh t Measurement s
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Pag e 3 2 o f4 6

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CropScienc e US004 6 MRI D 32539 , 12662 1

BAYE R A

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T PU R TOXIKOLOGI E Octobe r 24 , 197 8 

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## Bayer

CropScienc e US004 6 MRI D 8902 3

TERATOGENICIT Y TES T O F ME B 644 7 I NPREGNAN T RAT S

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St . Mariann e Universit ySchoo l o f Medicine , Kowosaki , Konagaw a Prefecture , Japa n

* Professo r emeritu s * * Assistan t Professo r

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Pag e 3 7 o f4 6

## Bayer

CropScienc e US004 6 MRI D 14933 6 \



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### Timoth yM . Unge r Danie l Va n Goethe m Thoma s E . Shellenberge r

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FINA L REPORT Augus t 31 , 198 2 MR I Projec t No . 7272- B

BC324/MO-02-00557 4 Fo r Moba yChemica l Corporatio n Environmenta l Healt hCente r Stilvell , Kansa s 6608 5

MIDWES TRESEARC H INSTITUT E '2 5 VOLKE P BOUUEVAHO . KANSA S CITY . MISSOUR I 5411 0 •81 6 753-760 0 Pag e 3 8 o f4 6

## Bayer

CropScienc e US004 6 MRID 15507 5 BAYE R A G INSTITUT E O F TOXICOLOG Y Wuppertal-Elberfeld , 30.5.198 4 Repor t no. : 1271 2

MEB . (Triadimefon ) TWQ-GENERATIO H STUD Y WIT H RAT S (Supplementar ystudy )

b y Dr . R . Eibe n Appendix ; Histopathologioa l repor t b yLif e Scienc e Research , Englan d

Unti l the yar e published , th e finding s containe d i n thi s repor t ma yonl yb e use d wit hth e approva l o f BAYE R AC . Furthe r reproductio n o f al l o r par t o f thi s repor t i s no t permissible . 12712 1 M O 02-00561 2 Pag e 3 9 o f4 6

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### Pag e 4 0 o f 4 6

## Baye r

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MRI D 4144620 1 Repor t MTD014 9 Moba y5o9 6 Date ; Marc h22 , 199 0 US004 6

Teratolog

yStud yi n the Rabbi t wit hME B 644 7 (Triadimefon )



### Author s G.R . Clemen s an

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CropScienc e

MRI D 4208960 1 Supplementa l Submission t o EP AMRI D 4144620 1 Stud yTitl e Teratolog

yStud yi n the Rabbi t wit hME B 644 7 (Triadimefon )

Repor t No . HTD014 9 Dat e Octobe r 28 , 199 1 Dat a Requiremen t U.S . EPA-FIFR

A Sectio n 158.13 5 Guidelin e No . 83- 3 Author s G . R . Clemen s an

d R . B . Hartnage l Jr . Performin

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Laborator yProjec t I.D . Repor t No . 10002 6 US004 6

MTD0149/MO-02-00636 9 Pag e 1 o f

1 1 

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Pag e 4 2 o f4 6

## Bayer

CropScienc e US004 6 

### MRID 4215390 1 BAYE R A G FachbereichToxllcologi e Friedrich-Bbert-StraBs e 21 7 - 33 3 D-560 0 Wupperta l 1 Wuppertal i 2^.10 . Repor t no . 2'Q"?? *



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### b y or . E . Bombard an d 

### veterinaria nB . Schlld e

Stud y No. : T302762 6

20774/ M 

### Unti l the

yar e published , th e finding s containe d i n thi s repor t mayonl yb e used wit hth e approva l o f BAYE R AG , Furthe r reproduc tion o f al l o r par t o f thi s repor t i a prohibited .

Pag e 4 3 o f 4 6

## Bayer

CropScienc e US004 6 MRI D 4308930 1 BAYERA G Fachbereic

h Toxikologi e Friedrlch-Ebert-Str . 217-33 3 4209 6 Wupperta l Wuppertal , de n 13.12.199 3 Bericht-Nr. : 2077 4 A

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- ACompilatio n o f Historica l Dat a -

Dr . E . Sande r an d B . Schild e

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Prior t o publication , the finding s containe d i n thi s repor t ma y onl ybe use

d wit hthe approva l o f BAYE R AG . Furthe r reproduction o f al l o r par t o f thi s repor t i s no t permitted .



### Pag e 4 4 o f 4 6

## Baye r

CropScienc e
MRI D 4737710 1 BAYK R CROPSCIBNC E L P 07-1)72- 1 L STUD YTITL E

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Guidelin e42 6 (Draft ) Healt hCanad aPMR ADAC O No . 4.5.1 4

Author s L.P . Sheets , Ph.D. , R.G . Gilmore , B.S . Author , Patholog y Repor t H.E . Hoss , DVM , M.S . Stud y Completio nDat e Report : February 20 , 200 8 Sponso r Bayer A C Baye r CropScienc e Alfre dNobe l Str . 5 0 4078 9 Monhci m German y Tes t Facilit y Bayer CropScienc eL P Toxicolog y 1774 5 Sout hMetcal f

Avenu e Stilwell , Kansa s 66085-910 4 Stud y Numbe r 07-D72-I L Pag e Io f 117 9 M-297829-OM US004 6
Kcpn nNumbe r 20176 6

Pag e 4 5 o f4 6

# Bayer

CropScienc e

MRI D 4737710 1 supplemen t HAY'lift CROKSCUiNCI : I f .V IUD YWO-HTMI
US004 6

KH'OKTNI-MHH K 201766- 1 Supplementa l Submissio nTo : U S EP AMRJ D No . 4737710 1 STUD YTITL E ADevelopmenta l Neuroto.vidt y Stud y wit h Technica l Grad e Triadimefo ni nWista r Rat s Dat aRequiremen t U.S . EPA , OPPT S 870.6300 an dOEC D Tes t

Guidelin e42 6 (Draft ) HealthCanad aPMR ADAC O No . 4.5.1 4 Author s o r

Supplementa l Kepur t D . I.. . VanCoeihei nan dL . P . Sheet s Stud y Completio nDat e Origina lReport : Februar y 20 , 200 8 SuppliMiifniii lRepon : Jul y28 , 200 8

Sponsor Baye r A G Baye r CropScience . Alfre dNobe lSir . M. I 4078 9 Monhein i German y Test Facilit y Baye r CropScienc eL. P Toxicolog y 1774 5 Sout hMetcul f

Avenu e Stilwell , Kansa s 66085-9104 Stud y .'Numbe r 07-D72-II , I« f 1 6
M-297829-02- 1

Pag e 4 6 o f4 6