Document B5Y16G5y2oG1QKmRzV9KzVn28

AR22~1846 . }i sos: APILOT REPRODUCTION STUDY WITH THE MALLARD FINAL REPORT `WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER: 454-105 3M LAB REQUEST NO. U2723 FIFRA Guideline 714 AUTHORS: SJReaoaynumPo.Bn:GdaBlLel.aavVgeheressrkoven STUDY INITIATION DATE: February 28, 2000 STUDY COMPLETION DATE: December 15, 2003 -- Envi53r3o5MnBmCaoerhnptoArLavatebionornesiory Spa, Manisa $5106 Wildlife International, Ltd. 5558 Commerce Drive `Easto#n,1M0ar2y2l.a8n6d0201601 Pat 100122 | | GNO3=9 Wildlife International, Ltd. Project Number 454-105 a GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT SPONSOR: 3M Corporation mE: PROS: APitRepraducion Suith d heMalylard WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER: 454-105 STUDY COMPLETION: December 18,2003 bythe TUSh.eEsmttudoynvmeanstcaolndPurcotteecdtiionncoAmpglciyan,ce40wiCtFhRGoPordtLa1b6o0r,a1t7oAruyPgruasctti1c9e8S%t;anOdEarCdDsaPsripsuiblleissheodf 3G8o5o0,dALganbeoruattroaryl PPrraocdtiucceio(nEBNuVeMaC,/1C0HAEuMgus(t981)98147,)waihdhJeapfoalnloMwAinFgFe,xce$p9tiNoonbs:San NotScaion No. protocolT(hWeisltuddeyIwtaesmcoanidoucntledLiudn.deromdyuluimpbeeprrot4o5c4o-l1s0.5)T,haendntihfeatpaoyrttiiocnlwpaorstcioonsnwedreucuocnnddteucretceodde umntwdeospeaOrrTae prowtaocla s (ExygewnilRhescaprachrstteudSyanuymbDeorcsto0r23-042 ad 023.063). Exygen Research reporteRdesseuplatrsaoelfyanalysesconductedbyExygenResearchforstudy pumbers 023-092and023-063are study uAmmbeenrdm0e2n3-t0s42mdumobnero2haavndt3heanSdpodnesvoira'tsisonign1aftourrtethoeEinxdyigcaetntReessearchihnagneaslywteicralapprportoovaeldfboyr he Spomor STUDY DIRECTOR: , SSeanMnor?lBGaollgfag-,herADvibanlTobxieconlogy DATpEale SPONSOR'S REPRESENTATIVE TL NewsieAd. BAT3E A 104 CNO350 Wildlife International, Ltd. ProjectNumber 454-105 a GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT SPONSOR: 3M Carporation TILE. PROS: APilot ReproSudduywicthtteiMaolnad `WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER: 454-105 : STUDY COMPLETION: December 18,2003 bGyootdheTLUahSb.oersaEttnouvridyryowPnramacestncitocaneldu(PcrEtoNetdeVci/tMnioCcn/oAmCpgHlEieMn,sc(9e480v)Cit1Fh7RGPosoaaddrLJa1a6bp0oa,rns1Mt7ArAFyuPFgr,uasct$ti0c9eN8oS5tb;aSnOadEnaCrNDdo,stisPfirpcauitbilsoinesNhoeo.df 5850, Agieulnal Prodcion Buea, 10 August 1984 ' STUDY DIRECTOR: SenLiGaallnagher8. Falla Senior Biolog, Avian Toxicology SPONSOR'S REPRESENTATIVE BATE los sr -- -- GRO3CA Wildlife International, Ltd. PrsectNumber 454-105 ae QUALITY ASSURANCE STATEMENT GbyootdheL"UaTSbh.oirsaBstaotvruiydryPowrnaamcseiencxaealm(PiErnoNeteVdcIftoiCorIncCoAmgHepEnlcMsyn,(c5e48w0)iC1t7Fh)GRonpoadrdtJL1aa6bp0o,arM1t7oArFAyuFPg,ruasc9tti1cN9e8So5t;aanO,dEarCNdDosdsiPscrpaiussbiilopinessNheoodf 3t85e0,sAygrdiicnugslwerroedurcepioortnedBtroetahse,S1t0udAyuDgiursetct1o9r844.5dLTahbodrastreyosfMalnladgeimesntawneriensapecotlioonwsan he Acviry PTroesptaSnbtsotnace Semple repntion DicPruion Blood Collection ADnraiRyeapoDnus DDirokgRiecpuoDnuns FDirasRseepos DATECONDUCTED ~~ STUDYDDAITREECRTEOPRORTMEADNTAO:GEMENT Feary 20,2000 Mirch1,200 March, 2000 Age 11,2000 Fer 25,200 Mar 1,200 Much, 2000 Ape 11,2000 Ma1, 2h000 Mach 1, 2000 Mura, 200 Apel12,2000 Tuy16,11,20829,0000 Janay20,2000 mFaerayr2y1331.,2000 Februar 200 Janay31,2008 Machiz, 200 December13,1003 Deceaber 32008 Decem18b,2e00r3 Allnspectons wer dybasedness bros ted. QKuiimblatnyyAsosurxancie Representative BATE(2-13-03 CROSSE YWEilLdlLifEeHInTtNerAnCatOioUnRal,, TLeHde. eee ProjS ectNumm ber45s 4-105 a REPORT APPROVAL SPONSOR: 3MCorporation TLE BFOS: A PilotReproSduduywicthttheiMaollanrd `WILDLIFE INTERNATIONAL, LTD. PROJECTNUMBER: 454-105 MLAB REQUESTNO: Uz723 , STUDY DIRECTOR: SeaL n Gan llagher . SeniorBioAlvioanTgos,co . BATE| da 7. CHEMISTRYPRINCIPALINVESTIGATOR: Scien,AnalaytnicaHloCvehne,mPiDs. wll AE MANAGEMENT: T_ ou --Bers ---- DireAvicanTtoxoicorlo,gy ilidAB.lNiixon, PhB. p DiroefChcemtisoty BATE reliefs BATE7 600383 Wiillddlliyfee IInmteerrnnaattiioonnaall,,LLttdd.. 5 TAOFBCONLTENETS PProjeect Neumbwer4e54-1e05 OVENS... Test SubansdTtteraalSnWBc.e AT Hovsing 0dEnViromestalConditions. 13 AAGGUU BBoId00ydWCeOigIhCaOnd.F.o.s Comumpion. cee 1166 A8d8uCNeOcrIopecy0ncdSiOTMioEeSsnCalerorrtion rr 1166 OfBlop od4drTesn sColeg cion ee 18 RUS 08 DIEU... 20 AdMultoBodyraWediCglahitnicaanlldOFBoSoeedrCvsOsiOoSnUsIZ0n. rerer 2121 TUES ABN 23 600351 Wildlife International, Ltd. ProjectNumber 454-105 "6 `TOAPFAGCBEO2NLTENETS TABLES Table 1. MFeraomnMoeMaaslularreddPCotoRnEpcIeOU(npCpSttmR.irYo)noaftPv FOiSionAnviase n Diet n 21 Table 2.PMielaontRAduEltpBordyOWSeAtiugUdhytCW(gi)ttfhioPRmoOSan.M.al.lard 28 Table 3. MBeiaontFReeedpCroodSntusyc(gWtubitiihdm/odapny)ftrPomiaFMoalOanrdS..ore 29 Table 4. SPiluotmRemporofadGurcrtoiyosnPSattuhdoylowgiictahlPOFbOsSer,vAadtuilotnsBirrdsaEouMhaalnm liazredd U6 Weeks20d Tes TErMUOREO....vrs 30 Table 5. M`eMaalnlaErdggPPilrootdRucteiopn(rEgosSLtdauidu/yHwecintthaPnidROESgo.g.sn.HenDay)froma 3. Table 6. SruommmaarMyaolflRaerpdrPoidloutcRtievePperrfoormdaSnctuey(cEgwtigtshSietofinomWePeFkO5S) .....vccr 32. "Table 7. Meraonm BaoMdalylWaerdiPgihlto(t8)RoefHpatcrliongsSdtaunuddyScwuirttvhiivinogOnffspPrFOi.n.g. 33 Table 8. MeanLiverWeights 5)from a MallardPilotReproduction StuyWith POS vss 3 | i CRO335 _-- mm Wildlife InH ternatO ional, Ltd. eee Project NumbA er 454-105 ee a. `TOAPFAGCBEOSNLTENETS APPENDICES ADEUX 1. CreOfaABYtSS. 35 Appendix 11. Dictand SupplementFormulations... 38. Appendix IV. TheAnaoflPEOySinsAWiADDsIE vv 40 APP V. DIOTtGLaYOOl rrr 54. Appendix VI. Appendix VIL `Appendix VIL Appendix IX. ARdulEt PBodTy WOSeiTGghtI(Wgi)CtfhrPHomR0aMOaDllSar.d P.ilo.t 55. PFieloetdRCoensupmprtioondS(tbuudicynWtidtih)oronm aMPaROlSl.ard 59 MdailvlirdduaPliGlrotosRsPepartohSdotOublysceoritvatgtihiioPonFcOnSfarolmv a .63 HSIOPSHOORYREPO... 61 Appendix X. EMgagllP ardr Piloo tR(ed egpgsu rlaoic dhdSet nut/wyci eewtkio )tfihrPoon RmOanS cc 106. Appendix XL ReMpalrloadrudctPiilvoetRPeerfoprmrancoeSdbtyuPudeyncWfirtotmhiaPFoOSn v.10 Appendix XIL MPielaotnROffespprirngBooddSy6udWycweitthiigofrnhom taMaslPROlS.)a. rd 16 Appendix XIII AdRulEtLPiveTr WOSeiIgGhtUW(ig)lfChrPoHRmOaSO.Ma.l.lardPilot 17 Appendix XIV. POAffOsprRinegLipverrWoeSigduhdtuy5)wcfirttoPmiRaOMoSalnlard evr 19 i| Appendin. XV. ChatoSntudgyPreotocsol. srmm-- || Appendix XVI. Personnel IaVONed ia SAY ...eceersususnsssmmsnssrsnn 122. | i R036 rem ------------------ Wildlife InternaO tionA al, Ltd. eee 5 SUMMARY STUDY: PFOS: A Plo Reproduction Stdy witheMtallhard Projecta Numbt er454t -105 SPONSOR: 3M Corporation WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER: 454-105 TEST DATES: ESxtpuedryiImneinttiaaltSiountF-ebFreubarriya2r5y,2290,020000 ABdioulogTiecarlmTienaromninAapteilao1lny&26J,2l00104,2000 TEST ANIMALS: Mallard (Anasplatyrhynchos) AGE TEST ANIMALS: Approximately 27 wesks ofges the inationofthe st SOURCE TEST ANIMALS: W1h3itWaisnhginWgitnogsn,Sotce.t UHSaAno,veIr, 61041-0509 NOMINAL TEST CONCENTRATIONS: 0,18, 62,0d 17.6 ppm. RESULTS: iMacltTloaorrrwdaenewekarsdea,ieiTxophnoeaslcedo1n3toowlPeeFgkOrsSo.uNaptaodnirdeets1at7rm.yc6pnctopnrmceelanaittr.eadetimtoonsrooafulp,eswe1rw8ee,rm6ei2csbansiedrnve1ed7d.ao6 pcnpytm : 3oif.tthhteesctigocnnosnccoeefntntortxairtciaiottnysie.toshtanAtesLdm.haNyeoh1oa7vv.e6ertpbsepiemgnn8as.o1lfexdot ccoontrceewnseierranettino.no,tTeadhesarientghwler1be8enno1waa6s.p2rppeprsde reatment lated cfict on bady weigh amongstmalesand foale inth 18 ed 6.2 PW1TPee8Mema,e.2is6. otsi2repsath1er7ceo6n1nc7pe.r6pnctmpraapmatmeii.nointrs.c.laeTitshmeteedngrtrecogwurlpeousrtpehstma.nemaTfaohnyoebdraoecvdwoeyenwrbeseieungnohmeatoplapooasefrse8enfstoatepmrsoeotndmtgoemcn.aitloanecsTnlahtsnhredede omffiisccertleoscvotenesdacinuyostofuntrhfeovrerasedipuorldotdaumchatiligevhseeripnrianhcmeiedte17nr.cs6e pomfep(dmsuacri.d.rgroHuip1.setgoeWpsahitohinolahonegdicpocobassmeiremvaasttiiuoonens 1um0paPoyFnObheSeiinrnceistduhelntdtsioaclftttfoiotsrsesistxiumwdeyen,te,hkeaswnra-esoa6bmse2enrptvpcemda-t<ief.edcetfcfoentccenoturadtinoontfboe pmraelcliaurddeds.ExBpaosseedd GRO3356Y7 --_--_----,--,--,--,--_---- = Wildlife International, Ltd. Prsjct Number 454-105 5 INTRODUCTION This study Itermatonal, Lt. was conducted avian toxicology by Wife Intemational, Ld. for fcilty in Eston, Mayland 21601. 3M The Corporation af he Willie biological orion ofthe test was conducted from February 29,2000 uni Jy 2, 2000, Raw ota generated at Wilde Iteoationsl, WLiidldaned Iantceompatyioonftalh,eLdfi.nailter.epoBritoalrogicfalledspuencdiemrenPsroajrectstNourematb34M54C-o1rp0o5raitniaornc,hSiv.esPalo,caMteidnnoensotthae ssiz. omECTIVES ofdieta"rTyheeoxbpjoescutrievetooftthheipsosttausdysiwmass0t oefvaPleufalteuotrheooecfafentesSuuplofonntihceAacdiudlt(hMraelalflredr(rdenlfearteydrfohy3cFhOoSs)) ocvoenrsuamppetriioondwofeprpevraluoatxedi. mI6naawdtedieetkiolsno,yrthe19efwfeeetkss.of EafdfuelcttsexopnosaudruelttohPeFaOl,Sboondtyhweeuigmhbte,ranodffeogogds elaxiadm,infearttiilointy,ofemseblreycotedvitaibislsiuteys,ahnadtcahnaabliylsiteys oafndbloofofdsparnidngtissusruveivsaalmpwleerseweevarluasltseod.use~dHitsoteovpaatlhuoaltogitcahle effects upon adults expose to PFOS and 10 her opr EXPERIMENTAL DESIGN Mallard (20 male and 20 mle) were randomly disribted ino one control group and thre rcament groups. Th test concentrutions were selected in consullion with he Sponsor, based upon the results ofa LCSO study (Wildlife Intemational, Ltd. Project Number 454-102) and additional toxicity | info ai. rmation provided by the Following experimental Sponsor. start he The test original materia test was concentrations selected were 0, reanalyzed nd the final pricy 2, w 7 and20ppm as lower than | orginaly reported. Therefore, the actual nominal est concentration were 0, 1.6, 62 an 17.6 pom a. PROS Treatment Groups Group NominalgpComncaein)tration PGormsouppor T 2 Cont) 0 1s 5 5 3 ` 62 116 5 s MaBliersdspeFcemPaelnes 11 11 11 i! 000353 Wildlife International, Ltd. Project Number454-105 "0Eachtreatanmdceonntotlgroupcontained ivpaisofbirdswithonemaleaodon femal per pen. Thee treatmentgroupswerefddietscontainingcither 18, 62,or 17.6ppm ai. of PROS. The contol groupwas fedditcomparabletoth rcamentgroups,butwithouttheadditionofthetest substance. Adultmallnds were exposed to PFOS at pominal dietary concentrations of 8, 62 and 17.6 ppm ai. foarperiodof6 weeks. Acontrol group, fdnon-reateddie,wasmaintainedconcurealywiththe treatmentgroups. Eachtesmentandthecontolgroupconsistedof fivepaisof birds,housedwithone maleand onefemaleper pen. At the endofWeek 6,adultbids in the 18 aad 62ppm a. test concentrations wer cutbanized and subjected to gross ecropsy. Test birds in the control group and 17.6 ppma. testmentgroupwere maintainoendtheapproprisedietsuntth beginningofWeck 20of thetest,atwhic imetheywere also cubanizedand subjected 1grossnecropsy. Effonealct htealsth, body weight, and feed consumption were evaluated as well s effects upon egg production, embryo viability, btcabilty and offspring survival forall test concentrations. The dul binds were observed daily for morality, sboormal behavior and signsoftoxicity. Adult body weights were measured at tet initiation, on Weeks 2, 4,6, 8, 10and 1, and at alt termination. Feedconsumptionfor achpen wasmeasuredover sevendayperiodeachweekthroughoutthe est. Nectopies were performed an all adult birds and on 10 offspring from each est concentration. Liver weighs were recorded for all necopsied bins, Liver, bile and blood (when avaiable) and fester samples were collected st the timeofecrapeyfo possible analysis. Addional samples of liver, bri, Kidaey, gonad, proventriculus, gall bidder, adipose tissu, and bursof fbricus (when availabl) were fixed in 10% buffered formalin for isopatbologicl examination During the test, the number of cggs lad in cach pen was recorded (0 value egg production. Eggs liinWedeks 1, and 6ofthetestwerecollctaendseparatedintoshell,shel membranyeol,k and albumen and sored fozenfo posible analysis. Es id uring Weeks 5 nd 6 (Days 31 10.37) of the test were collected and so incubation. Embryo viability, hachabily, batching healt an survvability were examined. COED Wildlife International, Ltd. ProjectNumber 454-105 ER MATERIALS AND METHODS Th studywasconductedsccadin t theproceduresoulined intheprtoca,"PFOS: A Pilot Reproduction Sudy with the Mallard. The protocol was based on procedures outlined in the Environ meatal Protection Agency's Registration GuidelinesPesticideAssessment Guidelines, FIFRA Subdivision . Hazard Evaluation: Wilde and Aquatic Organisms, Subsection 71-4 and the ASTM *Sundard Prac for Conducting Reproductive Studies with Avian Species" (1.2) Test Substance and Intern) Standard "The tet substance, PROS, was received from 3M Corporation on October 29, 1998 aad vas signed Wile nermtonal, Lid. identificationumber 4675 upon ep, Thtestsubstance was a white powder and was identified as: FC-95; Lo 217, The test material hd a ariginl reported purity of 98.9%snd had expiredattetime ofexperimentalsar. Anseyofth texmaterial fe xpermental tact indicated pity of 0.49%. The ial ssayofth test material indicat o pury of 869% nd `expiration dateof August 31, 2006 (Appendix I). The test substance was held under ambient condition in sluobcskteadnscteoriangteheatdtihet WwielrdeliafdejIunstteerdnattoio1na0l0,%Ltadc.tifvaceiliintgireesdiinenEtasbtaosne,dMuarpyolnantdh. oCroingciennatrraetpoirotnesodfpturhietyteosft 98.9%. Dietary concentrations are expresseda partspermilion acive ngrdicat(ppm a.) he di bpes onthe finarppourirofe86.d9%. "Th itema standard, 4 PFOS, was recived from 3M Corporation on July 2, 1998 ad was signed WildifeIntemaionl, Lid dentificaton umber 4526 upon recip. The ineral standard was o rons seri denied ss. 1, 1H, 2, 2H Pecloroctne Sufnic Acid; CAS No 27619:972. "he otal standard was bel unde ambient conditions in locked sorge tthe Wife nterasional, Ltd. facilities in Easton, Maryland. Test Organisms Fifty-one (24 male aad 27 females) penrsred mallards were purchased from Whiting Wings, Inc, 113 Washingion See, Hasovr, IL 61041-0509, USA. At hestart of selimaio, the mallcds appeared beslthy nd were pheaotpiclly indisinguishable from wild type. The birds were from the amebatch,approchin theirfisbrecdin seasonandhdnotbousdinanyprevious sing.ALthe sur of acclimation, random number generating, anton in 3 spreadsbest program was used to randomize pen assignment fo cach bid. Immedisly peor t test nitaton, all potential study binds 0370 Wildlife International, Ltd. Project Number 454-105 ne were examined for physical injuries and general helt. Birds that id nt appear eal, iter ue to injryo inability to acclimate to borstory conditions, were outside he weight range or the test, were excluded from the study. Al birds were approximately 27 weeksofage at test iniintion (frst day of exposuretotest die)and rangedinweight from 788 to 1258grams atest nation.Se ofthebirds was determined by a visual examinationofthe plumage. Identincation Adu bids wer identified by individual lg band, each pen ws deniid with unique number, and groupsofpens were identified by projec numbersnd concentration. Durtihentesgt, the number of eggs idin eachpenwa recordedto evaluate eggproduction. All eggscollfoerscamptliengdor incubationweremarkedwiththepen numberusing apermanentink markingpenforidentification. Hatelings were intisly identified by wing bands o hat hey cold be traced o hele parol pen of origin, Wing bands wer removed and th offspring were banded ith Ie bands st sproximatly fous weeksofage. Avian Feed and Water Al adult birds and their offpring wre given feed and watera iin during acclimation and testing, The basa dit fed to both adults and offpring was formulated to Wildife Intemational, Li specifications by Agway Ic. (Appendix I, Table 1). Th basa ton conaiaed atlas 27% protein and 2.5% ft,and nomorethan 5% be. "The basa dic contained approximately 1.1% calcium, derived from feedstuffs nd he 09% Jimeston used i he formulationof he basal diet by Agway. While his evel ofclu is sufcea or growth and maitessnce rations, additonal ese i require in the ron of breeding birds for eg shell formation, Therefore, an dion 5% (wie)of lesion (spproximaely 38.5% Co) was added to the basal diet for the adults. This raised the calcium level in th dict for the brecding birds 10 approximately 35, slighly shove th minimum recommended for ual 2.3%)sad mallard (275%) GO). Offpring received basal dit without tet substance and without the addition of 5% supplemental limestone. Water was supplid by the townofEaston public wate supply. Neither the adults nor offpring secsved any form ofmedication inte feed din the test. Feed and water were alyzed pesiodically in accordance with Wiki ntrmtinal, Lid. Standard Operating Procedures. CROTIR Wildlife International, Ltd. Project Number 454-105 nDiet Preparation st dietswerepreparedbymixing PFOS to premixthet wasusdforweeklypreparationof hefinal diet. ControldictandcachratedditweepreparedweeklybeginningouFebruary28,2000 andpresented othe birdson Tuesdayofeachweek.Diearyconcewnerteardjuastetdfiorpourintyosf hetest substanceandare presentedaspartspermilion active ingre(dppime8n.t) Detoiftlhe weeklypreparstionoftestandcontol diaersthowsn in Appendi IL Diet SaHmopmloignge.neity ofthe test substance inthe ictwas evalbuycaoltleind six samfrpomleaechsof he resteddies ndamesamplefomthecontrol dieton Day 0of Week 1. Sawmerepcolllecteedfrsom the tp, middle and bottomofthe Jf and right sections otfh mixing vessel. Control and treatmeat rou diet samples wle s collr ectedfom the eed troughs onDay 7of Weck1 0ssess stabiolfitthye {est substance unde actual tet conditions, Addionlly, a sample was collected fom the control snd ratmentgroupdiesduring Week 6 ofth est fo measureveriytestconccttions. Theditsamples were sored frozen or transfered immedistely the Wiki Intemational, Lu. analytical chemistry facil for analysis. `Analyt"iTcheamleMtehtohdoduedforth analoyfsPRiOsS in svi dict ws baseduponmethodologydevelopedat Wild Iterations, Lid. and ented "Analytical Method Verificationfor the DeterminationofPFOS in Avian Diet (Wide Intemational, Li. Pojct No. 434C-110), soluio`nSamcpolensniwenrge e0xtr0ac1tmedg0w1iH0t,h me1tHh,an2oHl, a2ndH diPleurtleudoionroact5a0n%e methanol Sulfoic : 50% NANOpure water Acid (4H PFOS; imermal standard) and 0.05% formic acid (vi) so that they fell within the calibration range of the PFOS methodology. A melbod flow chats proviniApdpeenddix IV, Fire 1. ConcentorfaPtFOiSionnthse standards and cxracts of the samples were detemind by reverse-phase high performance liguid chromatography using witPackard Model 1100 High PerformanceLiquidChromatogaph(PLC) wih PerkinElmer SCIEX API 100LC Mass Spectometr equipped with a PerkinElmer TurbolonSpray ion column (50 mm 2 mm source. HPLC separations 1D., 34am patle were size). achieved using a The instrument Keystone Betasil Cys analytical parameters are summarized in Appendix IV, Table 1 600372 Wildlife International, Ltd. Projct Number 454-105 "ae conniCanlgibra0t0i1on00stmagndards4HoPfFOPS F(OeSrpmraelparsedanindear5.0%anmdeth0a0no5l%: 5f0%oNcANsOcpdur(eVY)w,aterragsionlugtioinn concenuton from 0.000439 to 000439 mg aL (Week 1, Day 0)o 0.00351 to 0.00439 mg ai. s(aWmeeeka1n,dmDoasy7p, Wreeok 6m,piDeaanyk r0eaesnapdotntsheefsoarmPplerFew-exstOracltiSizoendsetto),mwoneirteoarnPalFyOzSeidwnilthctahleibsraamtipolne,sq.ualTihtey control, nd study samples No stmt was made to quantify PFOS on the bassof individual someric cstoamnpdoanredn)tsv.erLsuisneatrherergersepsescitoinveequcaotinocnesnwretroengenraetriaotsed(uPsRiOnSg pe: atk raraealresstpaonndsaerrda)tioosf(tPhFeOScairntseronanl sandars. An example ofa calibration curve i prscated in Appendix IV, Firs 2. The cononirsion oftest substanceinthesampleswas determined by substithtepuetakianrega responsratiosofthe samplesintothe aplicabe liner repesion equation. Typiclionchromatograms oflow and highevel clibraon standaradrse shown in Appendix IV, Figure3s und 4, respectively. Examples ofcalculations ae presentiednAppendix IV, Table 2. The method limi ofquantitation (L0Q) forthe Weck 1, Day 0 snalysis ws se at 079 ppm icalluciuloantead casothoefptrhoedmucattoifxthlealokwseasmtpclaleib(r2at0i0on0LsItKagn)d.arTdhaenamlyzeed (0L.t0O0Q04fh3o9rmtgoheaW.ied./eL)kan1d, Dthaeyov7ernaldl Week 6, Da0y analyses and the sample re-cxacion set was set at 41 ppm a... calleda the productofthe lowest sanded alyzed (0.000351 mg 3.1) antheovers dion toofthemati lank samples (4000 LIKg. ExampleofclcsueprlesenitedionApnpensdix IV, Table 2. Along with he sample analyses, four mix blacks were analyzed to determine possible interference. No interferences were observe at or above the LOG during the. sample amlys (Appendix IV, Tble 3). A typical on chromstogram of s mati blak i presented in Apendis IV, Figs. Avian dit samples were orf at 0379, 879, 3nd 220 pm a. (Weck 1, Day 0) or 176,879 and 22.0 ppm a. (Week 1, Day 7; Week 6, Day 0 and the sample re-extracton sc) and analyzed concurently with the samples to determine he mean procedural recovery. The method yielded mesa procedural recoveries of 105%, 108%, 107% and 1025. These vales comespond to exch sample st amlyzed or reanalyzed during the defi study (Appendix IV, Tobie 3). Sample measured GOS Wildlife International, Ltd. Project Number 454-105 "is concentrations were not comecied for the respective mean procedural recovery oftht sample set. A ypial on chromatogramof mati foriication i presented in Appendix IV, Figur . Housing and Environmental Conditions Housingand usbandey practiceswereconducted 3 8 (0adhereto te guidelines cxablihed by {be Nasional Reseach Council (4). The adlt binds were housed indoors in batesofpens mance tured by Safeguard Products nc. (Model No. S355), messing approximately 75X 90X 45 cm high Thepeas wereconstructed ofvinyh<ostedwiremesh. A diagramoftetes Iayou ispresetedin Appendix. Bachpenwasequippedwith fstrough. Weekly,suficient fodforte dingperiodvas placed the roughforcachpennd resentedtotebits. Duin tefein periodadios eed was weighed snd added1 th troughs a ceded. Wter was suppliedbyigpe-ype vateers. Only birds associated with tis study were maintained fn the study room in onder 0 void excessive dishubancs.The avers temperaturei th dot alld studyromdri the couofsbee est was 259 2 0.1C (SD) with sn average relive humidity of 48% + 17% (SD). The si banling ys inthestd oomwas desigaed to vent upto 15room sirvolumesevry hoursndreplacethen wit fresh air Thephotopericdintheadultmad room was maintainedby atim clock The photoperiod | d{hueribnigdasccrlciemvaetdiaonmaendantheotfessptrwoaxsim1a7thlyo2ou0f2rluigssht(p~e1r8d.a8y.tcoaindni)doefgulgllaicynianegt.ioTnhrporuogvhiodeudt btyheltces-t, | escent lights tht clocly proms oon-dy sunght Observations The ts is were aslimated t th flies and stdy pens fr spproximatly 4 wecks pri 0 inition of the test. Dring acclimation, al bits were abcrvd dal. Binds cxibiing sbmomal obebhsaevrivoerdodraidleybifloirtastiignngsophfysiiccailtiynojufribesnowremrealnobtehuasveidorf.orAtdhdeitteisotn.alDluyr,ienlg otfhfessptruidnyg,waellradoublstebrivredds dwaelrey from hating uni einaion. A record wes mained fall morales ad lial observations ros Wildlife International, Ltd. Project Number454-105 "16 Adult Body Weight nd Feed Consumption Adult body weights were measured at test nitston, on Weeks 2, 4, 6, 5, 10, 11 and at adult {ermination. Feed consumption for cach pen was mesured weedy throughout the test. Feed consumptionwas determinedbyweighingthefreshly filedfeederonDay 0,recording heamountofany addionaldiet addedduringtheweek,andweighingthe feederandremaining feedattheendofthe fecdingprod (Day7).Theamountof fodwasted bythbirds wsnotquantified,sicethe wastedfeed wasnormalysciere andmixedwith wateadexcreta Therefor, eedconsumispprtesicaodans anestimateoftotal feed consumption. Allremainingbidswee ethtahenediysgafeer dhe begining. ofWeek 20. Therefooorfeee,d consumption stiwasmcaltculaetedfo Week 20. Adult Blood Callcton Atthe endof Weck6,blood sampleswerecollectedfromall suvbirids,wvheinponsibgle. Following collection of blood sample, birds in the 1.8 and 6:2 ppm a. treament groups were euthanized. Additions blood sampleswere aso collected, when possible, rom birds hecontol group andthe 17.6ppm .. ncatment group prito euthanasiaatthebeginning of Week 20. All blood samples were separated ino serum and hemacyteslpltelets, stored frozen and shipped to Centre Analytical Laborstris In. for posible alysis. Adult Necropsy and Tissue Collection A the conclusionofthe exposure period, all suviving adult birds wer eutbanizd by cericl dislocation, necropsied, and stored fiozen, Al th time of necropsy, tissues wer collected for histopathologic examination 0 and 17.6 ppm .. groups oly) and pasile analyses. When available, samples of gall bidder, iver, proventriculus, kidneys, brain, gonads, bursa of Fabricius and adipose tissuewerfixed in 10% buffered formalin. Histology samwperelsheippsed fo EPL in Herndon, VA for Histopathology. When availble, samples of bil and lvr wer sired fiozen and shipped to Centre Analytical Laboratories, nc. fo possible analysis. Any remaining issue not iedforhistopatholaongdy fenher samples woere fe aze for ptential analysis. Egg Collection and Storage Eggs id during a seven day period beginning on the second day of Week ofth fet were llcted daily rom test pens and stored in a cold oom until incubation. Eggs to be incubated were washed reduc the posibityofpathogen contamination before soin them in the cold room. Eggs GOO Wildlife International, Ltd. Project Number 454-103 nallt or the fstfourdays were washed by and prtoitoorra. The remaining ogg were washed in commercial og washer(Kul Egg Washer) withachrisbased det(eKurlgSupeernCDt). Wate ithwasher waswarmed 0 approxima4t5elCy.Th eggs were placedi thwashwaterandsoe for approximately 15 seconds. The washer's circuliion motor vas the fumed on for appoximatcly ee mints. The eggs were removed fom the washer, allowed 1 ool to approximately room empnedrirnsedawithtehuwaeter.The codroomwasmatin a mesatemperature of 13.1 & 01C (SD) with 8 mean relative humidity of approximately 75% + 4% (SD). Groups of eggs were denbyaialephabdet ot cod. Alleggs aidduringtheweekwere consideredas on ok. Candiiag and Incubation Atth endofthe esky irval, leggswereremovedfomthecoldroom, countedandcandied wCirtachkeadoSpreeadbKnoirnmgal(eMgogdsewlerNeo.rec3o2r)edgegds-ncadnddilsicnagrldeadm.p todetectcggshell cracksor abnormaleggs. Al cgesnodiscardedwere laced i aPersieIncubator(ModelNo.SP20. Ihe incubator {oetmpertrewasmaintainedatam average 37.54 00C (SD)withan averweatbgueb empersur of 30.6: 0.1C (SD) relative humidity ofapproximately 60%). Theicuwabseqsuipptedwoit pulsator fin and blades that produced a mild beating si movement designed to eliminate intacebinet {temperatuarned humidity variation durin iocubsion. In ordertoprevent adheof stheiemobrynoto the "hil mambrsan he ncubtos was iso quipped with fa tomatic S55 otton device, designed to ott the ogg from 50 off of veal i one dieton to 50 offofvertical in he peste direction (total reofrotatiwoasn100)everytwohoursthroughDay24of incubation.Eggswerecandioedn Day 14 ofocubsion todetermineembryo viabanidolnDiaty2y1 to determine embryo survival. Hatching and Brooding allowedOntoDahaytc2h.4ofiPnacuibratsiobna,sktehtesegcgosnswtreurcetepdlaocfedgiannaiPzeetderssitmeeelHawticrheerme(sMhodweelrNeo.veSdP-6tHo)kaenedp htclings scpartd by parental pen oforigin, Eggs were not rotated in the batchr, The average temperature in the hatching compartment was 37.2 + 0.0C (SD), and the average wet bulb temperature was raised 0 33.0.28C (SD) (relative humidity ofapproximately 77%). GneEs Wildlife International, Ltd. Project Number 454-10 "sAlbatclings, uahatche eggs, ndcg shellswereremovedfromthehtcheronDay 27or 28of incubation, The group body weight ofthe survivinghatchlingsby pen was determined. Ducliogs were wingbandedforidentificationbypeaof origin andthenroutinely housed aceonto heappropriate parental concenation groinubroopdinigpennsugnt approximately 3weeofkagse. All ducklings were moved to pens without supplemental heat, whee thy were housed pproximately 9 weeks. The duckling were fed unrated dit without the additoifo$%n supplemental limestone. Atapproximately 12 weeksofage, the average body weibygprhetapenofal survivin ofspringwasdeteined, and he bidswereetbaized with carbon dioxide and dispose ofby incineration. Thoseoffspringselected for blood and tissue sampling were sored fosen following necropsy and. er disposed of by incineration. Htchlings were housed in baresof brooding pens manufactured by Safeguard Products, ln. Eachpenmeasuredapproximately 62 X52 X 25.5 cmhigh. The walls, lorsandcelingsofexchpe werconstructed ofvinylcoated wire mesh. Thermostat inthe broodingcompartment ofeach pen were sto mainatetmpaeraitunreofppronimatly Cfrom the timeofhatchwnintgebidsweefive to seven daysofag, when the temperature was dusted to maitsna temperatureof approximately 29C. Brooding was discomtiaued once hatchlings were determined be of sufficient se o hermoregulat. The average ambient room temperature in the room housing offpring was 23.5 + 0.6-C (SD) with an average eltive humidityof 714 11% (SD). All ducklings were removed from brooding pens and rated peaswithoutsupplemental es a proximately 3weeksofage. Thpensewereethe same model used to house the adult bids. Th photoperiod for theoffpingwas mainaned by a time lock at 16 hooufligsht er day. Offspring Blood and Tissue Cllcton Pio to cuhansi of the offpring, blood samples were calcd from 10 offpring in cach reament group. All blood samples were separated nto serum and hemacytesplatelets, ore frozen, and shippedtotheCentre Analytial Laboratori, nc. fopssible analyses. Additonal, tissues from he 10 ffping in cach group were calcd for bisopathologcal examination and analyses. Whea availabe, samples of gal bladder, live, proventriculus, kidneys, bri, gonads, bursa of Fabricius and adipose tissue were fixed in 10% buffered formalin and shipped to EPL in Herndon, VA for isopathology. When available, samples ofbile and iver issuewere stored foe and shippedo Cenire Analytical Laboratories, o. fr possible analysis. Ga 77 _-- 2 Wildlife International, Ltd. eePrS oject NuY mber45S 4-105 "19 Statistical Analyses stisicUaploynsicgonimfpiclaentondifoffertohecestsb,etawneeAnnaglryouspiss.ofDuVnanreiatn'csem(uAliNpOlVAc)ompwaarsispoenrpfroorcmeeddurteo (d5e,t6e)rmwianse suisgendif9iccanocmepoafrtheethobesterrveeed dirfefaeiremnecnets.meSaanmsplweiluhnitthsewceornettrhoeligndriovuipdumalepaansawnidtahisnseeaschheexpseariismeinctaall w5e0ruep,exeaxcmeipntedboudsyinagndDulninveertw'esigmhettshowdherfoeltlhoewisnagmparlcesiwnaeitswqauasreterooitndiavnisdfuoalrmbaintdo.n.PerTcewnotagseetsdaotaf ostnaltyistliocoalkeadnaaltybsoedsywweeriegchotnadunctefdeewdictohntshuembpotdiyondweaitgahftoanmd hfeedfrcsotns6uwmepeikisoonfdtahae. sOtnudeysoedofeavnaalluyasteesd ll thre restment treatment groups. The scond groups and examined data for set the of analysis only evaluated fll tion ofthe study, the conrol 20 weeks. and 17.6 Dunaet's ppm 8 mulple Sc5o0muppa.riTsohne psrtoucdeendtu'rse Twaessnowtacsonussiedderteodmsapkpersotpatriisstticoal ccoommpapraisrhoenscionnttohlosgrionuspta0nc8esswinhgelreeraesltymetnbte contolgroupand th 17.6ppm ai. tregraouptwemre ecomnparted. 1. Adul1t0B, o1d1,yWaenidgohttad--ulItndtievrimdiunaaltiboon.dy wSeaitgihsttiwcalscmoempaasruirseodnsatwteesrteinmiatidaetiobn,etWweeeeknst2h, 4c,on6t,ro8l, Sroup ad cach reatment group at cach weighing interval by sx fr he frst 6 weeks, In addon, siatisical comparisons were made between the control group and 17.6 ppm ai. rcagromupfeorwneskts 10,1, and20, 2. AdultFosdConsumption - Feedconsumptionexpress ss ramsoffedperbidperdaywas ceoxnatmrionleadndbyacphenrweeseeklnytdgurroiunpg ftohrewteeset.ks S1tatthisrtiocualg6hc.omIpnaraidsdoindosn,wearetimand!e cboemtpwaereinsotnhse 5. wermadebetweenthe control EggsLaid-The numberof eggs nd 17.6 ppm .. treatment grou forweeks?though 19 aidperemleper restmentgroup.Forthe evaluatioofn | 4. Viab`lreepEromdburcytiovsep-eTrhfoernmaunmcbee,rdoafltaivweasetmabkreynofsrdoemtewremeikned aetggDsasyet1.4 by candling. i 5. EggsCrackedofEggsLaid ~The numberof eggs determinedbycantod becl raci kedn divig ded i by the numberofeggslaid,perpen. i 6. ViabluemEbmebrroyfogsosfEsgtg,spSeetp~enT.he numberofembryosattheDay 14candlingwasdividedbythe |i 0378 -_-- Wildlife International, Ltd.eee 2 ProjetNNuOmber 454-105 eee 7. Live`3w-aWdeivcikdeEdmbbryytohse oufmVbiearbloefvEimabbrlyeoesmb--rTyohse,npuembpeerno.f live embryos attheDay 21 candling 8. Halchlings of3:WeskEmbryos --The numberofhachlingsremovedfomthe hacherwas divided bythe umboeflirve 3-weekembryos,perpen. 5. 1&:DeyOMSurvivorsofHotclings The numberof 14-dyoi survivorsws dividedby the 10. Hatc`lniunmgbseor fofshsatschSleintgs- per week, by pen. The umberofhatchlingswasdividedbythenumbofeegrgsset per week, by pen. 11. 14-DayOdSurvivor ofEseSet -The number of 14-Day old survivors was divided by the nuofmeggsbse peerwrek, bypen. 12. OffipBroidynWgei'gsh ~ The group body weights ofsuniving batchlngs aad 14-day old `survivors were measured by parental pen group. 13. of LiverW eight offspr -- Indi ing of vidual week liver 5. S awteiisgihctaslwceormepmaeraissuonrsedoaft daduulltteirvmeirnawteiiognhtasn dweart ethme atdeermibnya tsieoxn between the control and treatment groups. Juvenile liver weights were compared by test group, without regard to sex. RESULTS AND DISCUSSION 4. Adultmallardswere exposed to for periodof6 wesks A contol PFOS at nominal dietary concentrations of 8, 62 and 17.6 ppm group, fed non-restd dic, was mioained concurrently with the {rcamentgroup.Eachtresmeotandthecontrolgroupconsistedoffivepairs of birds,housedwithone male and onefemaleperpen. A the endof Weck 6, adultbids in th 1.8 and 62 ppm ai. text concentrations were culbanized ad subjected to gross pecopey. Test ids in he control group and 17.6pom ..raimentgroupweremainedonth apropriste diesualtebegining ofWeek20of he test, at which time they were ls euthanized and subjected fo gross necropsy. Analytica Results None ofthe control samples showed any indicationofthe presence ofthe est substance orofthe prescncofcelting substance a he characteris tein ime ofthe test substance (Table 1). Diet samples were collected from th 18, 62 and 17.6 ppm ai. test concenrtions om Week 1, Doy 0, and wer analyzed to evaluat the homogeneityofte est substance in th dit and o verify est substance concentrations. Meansand standard deviations forte thretestcomcenatons were1.8 4.0.13 ppm i, Gr0s7e _-- T Wilde life Intera national, Ltd.eee P ProjectN Number 454-105 ~~ a 600.65 ppm i. 228 176 1.46pm .. respectively. Thecosets ofvision were 72%, 11% and 83%, respectively. These vals reprsened 100, 97 and 100% of nominal concertos (Appendix IV, Tble 4). Semple callted during Week 6, Day 0 ofthe ett verify test sbsance concenaons forthe 1, 62and 17.6 ppm .. dishadmeansandsander devinionsof 20 0092 PPm a, 60: 0.67 ppm ai.20d 163 0.608ppm ai,respectively. Thecoeffiofcvariiaetionnwterse 4.6%, 11%and 36%respectively.Thesevaluesrepresented 11,97 and5% of ominalconcenrations t(AepmppeenrdiaxtuIfrVo,reT7adbalyes 5a)v.erAangaeldys8i9s%o,f7di%t asnadmp1le0s5%coolfltehcteedDfaoy0mvfaeleudeesrsfoarftehre b1e5i,ng6.b2eadda1a7m6bipepnmt ats concentrations, respectively (Appendix IV, Tule 6). Aypcaio chromaotfaotegstrsaampmle is showin Appendix IV, Figure 7. Mortalits and Clical Observations Noadultmoralesoccuredin thecontrol group or ianyofthe resimentgroups uring the coouftshtes, Severalbirdswernotweitdhbumbleoo (folesionasn)d ethosear result of penwearandorpeat aggressionduring the couse ofthe tes. Asingebid i te 62ppm2.3 cameo group was ned ith had ein, An nde cia ign, amencs,wasasocistd with he bumble oo Inte coped for 17.6 ppm. Seeing a neatmntgroup, test termini. th he She rom pen218 also xii arche hecekadndbecamerigidwhen body wemors ad rapid bliin and respiration durinthebledingprocess. Clisnignis acppaearled 0be igeebretdhe sess ofhain, "This henwas abserve tobe normal i appearance and behaviorprior fo et termination. All other bids atest ropa were normal nspperanc and behavior for hedrain fhe est. Adult BWohdeynWceoimgphatreadndtoFetehde Ccoonntsroulmpgtriouopn thre were 10 apparent treatment lied fects on alt body weights the 1.8 or 62 pm i. est concnirations and any ifrnces betwen he control group and 62 ppm a. test conenrmtion wee not staal signifian. Ther was a sastclly significant (<0.05) reduction febmodyaweilghtesi he 1. ppm a. fest conection romWeek 2(0 Weck4, However, is eduction wa no consistent ordos responsiveandwasnotconsidered came eaed. Thre were mean body weight ossamongmals and femalesinthe 17. ppm 3. came group ta may haveben elatedtotestment. Whencomptoathrecoentrdolgroup,mean male body weightforthe 600538 Wildlife International, Ltd. Project Number454-105 wae 17.6ppmai. estesgroupwessigificanly(p< 0.05 ower athe Week 10, 11and 20bodyweight cionntecrevnatlrsa.tiTohn.ereWwhasiatlhseorae wasignifaincaonvtelorsasolflosvoeframlelamneabnodbyodwyeiwgehitghftofrofremmalaelsesatitnthhee 1177..6 pp a. 6 ppm a.i. ttheset dairfefperreesnecnetfedroimn Tthaeblceon2t,roalnwainsdinvoitdusataltibsotdicyalwleyisgihgtnimfiecasnste(pme>n0.s05a)r. pMresaenntbdodiyn AwepipgehntdimxeaVsLu.rements Due 0 excesive wastage by some bids, fod consumption was variable between pens, However, When, compared to the control group, there appeared to be bo teamentrclated effets on feed consumption st the 15, 6.205 17.6 ppm a. test concentrations. Whi ter were statistically sigaificat (9 <0.09) iffencsbetween thecontrolgroupand he 17,6pm i. eamgrouep fonrWeteks 14, 16, 1,18 and 19, these differences wer due to pparet cress in fod consumption or th 17.6 ppa. group. Feed consumption measurements at the 17.6 ppma. fest concentration were consistently higher thanthecontrolgroupwith camespondingdereseinbody welght,sgesing heincreasewaste restoffeed wasiage. Meanfood consumption measurementsare shownin Table3,and food consumption measurements by penarested in Appendix VIL Gross Necropsy AC heendof Week 6(Day 42) al adultissinte 1.8 and 6.2pma. resentgroupswere cbanized and subjected o gross necropsy. Adult bids ite cosrl and 176 pe .. resent groups wseubrjeecmtaeidntoaingerdososn ntehceroappspyr.opriAatle fdiientdsinugnstilobtsheerbveegdinwneineg ocfonWseideekre2d0taonbdeweirnecidtehnetnaleuttohaenisztemdeanntd Necropsy findings ae reported in Table 4 and Appendix VIL Study offring were approximately 12 weeks of age tthe imeofcubanasia. A subsample of 10joven birds frcach tet group were suited 0 goss necropsy. Nesropsy reveled several birds in cach eaten lve with bumble fot (ot lesions). Adina, eal offspring inte 17.6 ppm. ratment group was ned with picked and broken ethers and an kent sppesanc. Inemaly te bird was noted with a reine yolk sa, remnant of te stl yolk sac. One mal offipringi the 17.6pp a.. reatment groupwasnoted with heposteriorportion ofthe left Kidney absent. Al findings abseved were considered10be incidental 0 testment. ronan Wildlife International, Ltd. Project Number 454-105 ne Histopathology Light microscopic examination was perfoanrsemleectedd tisbsyauboearsd seid puthlogit at Experimeatal Pathology Laboratories In. Herndon, VA. Sections of liver, brain, Kidoey, gonad, proventriculus, gall ladder, adipose sus, and burs of fabri (when avilable) wer collected fom adulttestbidsandfrom 10offspring (spproximately 12weeks okatctbansi)fomeachtest groupfor examination. No lesions considered possibly related to treatment were noted in liver, kidney, oprfosvpernitnrigcualusa,nygalol tbfhladdceor,ncoevaariy,onbsraienstaen.d buArdsdaonofalflabyr,icituhsereofwaedruelt2m0aleeinansd cfeomnasliedse,reodrothebier reatmentened noted in adipose suofdat frmales and offspringofboth sxcs, or i th estes of one offspring. When compared to th conirl rou, ses ofadult mals at te 17.6 pm .. tet conection exited igher incidence of fats consistent witpostreproduce hase regression. Adsl mals inthe 17.6 ppm. group ls cisnit ncrease edndd e ofadipose se microvesiulto, The increased incidence oftesticular regresion snd adipose tsue microvesiculton fo adult males in the 176 pm a. esment group may be incidental testmen, but resentrelated effet could not be scouted. The ll pathologyrepris provided in Appeni TX. Tissue Analysts "he analysisofth eg, bloc and sue samples collected uring th study were conducted by Exygen Research (formerly known a Centre Analytical Laboratories) an rs reported sparc. Blood and ver analytical resultar reported in "ExtroafcPottaissoiunm Pesfloroocaneslfoate fom Quail semim and quail live for analysis using HPLCElectrospray Mass Spectromety; Cente Sdy Number 341". The egg components analytical rents are reported in "Extraction of Potassium Perforoocianeslfonste rom Egg Membrane, Albumen, and Yolk for analysis using HPLC ElctosprayMass Spectrometry; Gare Sy Number 023-065", 00052 Wildlife International, Ltd. Project Number 454105 ER ReprodWuchtiivegRgesuplrtosduction was highly varisl among ess, here were 2 sppaearestoen elated ffs upon cg production at say of the concentrations tested. Any ifereces between the contol upaadth restmentgroupswereno sisal sigifican. Whencomtotpecoantrorlgreoup, {here vas eduction i egg production atthe 6.2 pm .. rcament evel. However, isreductionwas notdose responsiveandws notconsidered redotreatment. ig production tthe 1.8 and17.6pp 8. testconcentrations was comparsbetoorexceededthecontolgroup. Mea eggproductiboyn conceisnprteserntaeditniTabolen5. Individual eggproduction dataarepresentedinAppendixX. When compared tothe control group there were o ppt resent ete efects on embryo vail, batchabily, hcg helt and survivability at the 15, 62 or 176 ppm a3. levels. Any ferences between th control grop snd th reste groups we ot satistalysgificnt for any `parametermeasured. Whiletherewerefewer14-dayoldsurvivorsperbenproducedatthe1.8and 62 pm i. test cononrtion, hese rections were de to foe eggs bein set for cubation a these levels. Reproductive dat re summarized n Table 6. Reproductive dat or individual pens presented in Appendix XI. Offspring Body Weighs "hers were 0 apparent raimentseated effets upon the body weights ofhatchlings in any of he rstment groups and any diffienses rom the conrl group were ot sail sigaiicant. There wre al no spparet came elated ffs upon the body weight ofjvenle birds in the 18, 62 or 17.6pp a. rstmentgroups. Offpingbodyweighdaipresentedin Tab 7andAppendixXIL Liver WWehiegnhtcsompared to the contol group, there wee 50 appartment elated effects on dul iver weigh at ayofth concentrations etd. There ws iaisialy significant (p< 1.05) increase in mca female vr weight at the 18 and 6.2 ppm a. test concentrations. While man adult vr weights were higheramongmalesandfemalesatthe 1.8 32d6.2ppam. etsconcentwhreaoctomipaorendsto {he contol group, these diferencs were de othe timingofadult termination frthese reatment groups. The 18 ad 6.2pm a. testbidswere utbnizedst the ndof Wek 6 oftetet,during aperiod of ighog production, Thecontolgroupsnd 17.6pm :. ramengroup stbis wrecbaaizad {oe beginning of Week 20 ofth test by whic time eg production had declined dramatically. Ore [OEE Wildlife International, Ltd. Project Number 454-105 Ee would expec birds in fll reproductive condition, particularly females, to have igher levels of fat reserves in the liver and correspondingly higher liver weights. When compared othe control group, there were mo apparenttestment.related effects on juvenile ivr weight at anyofthe concentrations tested. `While there wasa sigaificant increase in mean offspring ivr weight forth 1.8 ppma. reameat group, th increase was no doseesponsive and appeared f be elated to slighty higher mean body weights for this group. Mean adult and offspring liver weights are presented in Table 8. Individual adult liver weights are presented in Appendix XL Individual offspring liver weights are presented. in Appendia XIV. CONCLUSION Mallards were exposed to PFOS atdietaryconcentrationsof0, 1, 6.2 and 17.6 ppm ai. for 6weeks.The control group and 17.6 ppam. est group were maintoantiesntdeicdtsfo an additonal 13 weeks. Notreatmentrelated mortalities were observeda anyoftheconcentrations tested. No overt signs of toxicity were noted at the 18 or 62 ppm ai. test concentrations. AX the 17.6 ppm ai. test concentration, single hen was noted with signs of toxicity that may have been related to treatment. "There were no apparentteste.relted effects on body weight among males and female i the 1.8 and 62ppm ai. test concentrations.Thereweremeanbodyweight losses among malesandfemailnethse 17.6 ppm .. testgroupthat mayhavebeen related to treatment.Therewere noapparent reatmentrelatedeffectsonfeedconsumpiionor eg production i the 1.8, 62or 17.6ppmai. treatment groups. There were no apparent estmentrelated effects on any of the reproductive parameters messured. Histopathologica examinationofselected issue revealed a higher incidence of testiculr regression and adipose issue microvesicultion for adult mals in the 17.6 ppm a. group. While these observations maybeincidental to treatment, atreatmentrelated effect could notbeprecluded. Based upon the results oftis study, the no-abserved-effct concentration or mallards exposed PEOS in the dietfo six weeks was 62 ppmai CrOsaL Wildlife International, Ltd. "25 REFERENCES ProjectNumber 454-105 | USSub.dvEinsviiornonEm,enHtaazlarPdrotEevcatliuoantioAng:ency.Wif19e82.andPesAtgicaitdee.AsOsresgsamennst,Gusiudbesleicnteiso,n.FI7F1R4A, EnvironmentalProtection Agency,Office ofPesticidePrograms. Washingln, D.C. 2 ARmeeprroidcuacnivSeocSiteutdyiesfowritThesAtviinagn SapnedcieM.aterAiSalTsM, St1a9n8d6a.rd ES1t0a6n2d:a8r6d. PrAanctuiacle fBoorokCoonfdAucStTinMg `Standards. Vol. 11.04, PhiladelphiPaA,. 15pp. 3 Me&rCoc,nkc. 1991.TheMerck VeterinaryMarsal. Mer&cCok. Relay,NJ. 1832pp. 4 NaWtaisonhailogRetsoDenaC,r.chNaCtoiuonncaillA.cad1e9m96y.PresGsu.ide12f5oprp.the Care and UseofLaboratory Animals. 5 DuWmmeitatC,toCohWl.. J1o9u5r5,.AmAer,MuSlaitpsl.eACsosmep,aSri:so1n0s96P-r1o1c2e1dure for Comparing Several Tretments 6 Doulnnett, CW. 1964. New TabforlMuletipsle CompawritihsaCoontnrosl. Biomet2r0i4c82s- 600345 i z sz $5 3s i A il Blo g If; sl f: | =r oan = aE i Eom ossoae ii |1i d ] i|i! 13} 1 JOEii 1ocho .3 .fR3 i [lisi] : GRO3H5G J as. ProjectNumbe 45-105 Pal sare 0 00 Ga gs Sof gs ge co age jl en co see i= gegen gee] Bees meas fy el gras oo ee| i IH if of Bl] 335 [5s] il fel gmae eess meme 2m ge I RERTRTI | oo gas|) ooo senag erza mraz vase |F grzs zogn gms |} Menara i Ps] v= -o sans sm gr mw 3s Hy EF Jil EE vs 2a emse aLnRaTsRvLrTzRa i IE fa] 25 25 sass sm ss Fu: iH | fof E5L9E8C% E ge gL s 5L 7 geEgsRoge gf fii 7 Jr it .21g i0i1 2 ol 60057 HH i 34 &i ] 52 i 9. sl es 0 sl se 0 ses 0 = as ol se 0 Sogn 0 af ae 0 og 0 i= z= 0 of gr 0 - gs 11 ~ ga 1 *| ss me - = sv | oes oss ol sx ze "| se zs - 8 @y I 3fo gr `Project Number454-105 0a 0 is 0 Em 0 we 0 gw 0 ge 0 ge ge 0 ge 0 ge 0 ze tt am is ms as 3d sf 25 | df ose ov | IH zn ge {i ms zm [fi 3 38 ih gr gs [14 sis 00355 PH nin iiPdHa, i Pesiiqqiiiiing --_----mmm Project Number454-105 ze LE [] 23 feof BIE ste a 3 2 em i i maa : { i zee nee i i 3 Eon eee i 5 droog 5i2 Bol F sz sr zz osT: oli es 2 3| see gR= 32s zac 3 ID H2iE of ol fBa-eenn eaz nsee xw2 avn g3 egeos i 1h HS | wf mem gev men geo Ti AdfaanenoaeonenSnoosan hgalg He fs He He 1H] CRO330 n- Project Number 454105 Tables SummaryofReproductivePerformance(EggsSetfom Week 5) rom aMalar PilotReprodStuudcywtithiPoFOnS, ReproductiveParame-- ter _ ConC dO_ pepma) l) NumberofReplictes s Eggi u Eggs Cracked Eggssat u Vise Embryos 2 Live 3:Week Embryos n Hachlings 2 Oftsprag Survivors n Egos Laden s Ofpring/en ` MeTiEpmpgmaeiiesiGlGo2eppmmal s s n 1 0 wv n n n n n u n un n ` 2 3 2 MeTiepmmaali s u 1 z n 2 1 n s 5 Eggs LaidHenDay I ost 03 os EgerCracked gas Laid 0) s . Viable EmbryosEggs Set 06) % 0 10 2 Liv3-WeckEmbyos/Vabe 04) 100 100 10 100 | Halchlings'-Weck Ebrycs (4) 2 0 10 || Offping Survivrsiaclings 06) 100 " 100 " HtchinguEiggs Se 9) ss 0 10 3 Ofpring SurvivorsEges Se 00) 5 " 10 " Hatchlings/Hen/Day 060 0.40 034 051 Ofsprng SovivorsHierDay 00 ox 03 os Difrecesever the rl700 nd he resin rnp werea ical gat (> 009) GOEL  k i Jgf!:oro22 os ii i H i il 2) = x 3 2 i ii ih = hh Iioo=li: siie of iHlLl ! COS . a kl: 8 5 eSlkle, .ror Hi i Pid it ws 2 A i li 3 3 5 ple8 3 oF | 8 8 Jez oe a 1 LEoocoiibososbi os | | i & | i | 1 2 iti! i ili Ls | iE ssiiig] - 6r030% --_-- 3s. 2 e WildlifeAInternationale , Ltd. e ProjectNuA mber 454-105 eee Appendix CeriofeAnatlyseis Centre Ansiytical Laboratories. Inc. INTERoIMlCeERnTIFOIFACw NAALYTSIES n Contre naysiLTarebaFro7C0OrS8ALRheaerms C3184 oe es HETE m ia L{1 iosnsmaemr { qarms, r :i ey ira Seaenh rEpmE = -- nee TT GE] lum iaon i Sgumser IVaiEadaFe FE ||bl|Ll eo EPaeEvmt EeaSsol|l| EM1|r anemaErwtoee coum fo R05 --_--nmmm Wildlife International, Ltd. --_ %- Project Number 454-105 ApTpeonedsix] Carifcte ofAnaya: o a en CentreAnalytical Laboratories, Inc. CoInNtrTeAEnRtIoMnCLaEbRroeTtnIsGonFROeIFlACeNaAA,LYT2S3IE0S180 DeoraAmir: a0 [a -- PRP ---- RemaDru 83008 Pty AmTfeo pio,e45p%sLCSopoi,onHO,RT ' T Roa tor n e =y 3.70 "Rtncc frm thers etcod my. aime I -------- "tret to cores0 dbsdc rinie on boSn anie mlsi, io ni osc taps e. = | ANTamn Te N iti a e | EO mec us os ton ifor GFSOKOWS) | wrkoscnt PAGonLienPreSts (CTR16) anno tes G03 --_--_--m-------- 3. Wildlife International, Ltd. Project Number 454-105 ApFpeonedsix] CanifcteofAnsys fe Centre Anatical Laboratories, inc eae Cr ri CIoNrTeAERnIMnCLEbRstiTnCIsOAFROIFtACnNA:ALYT1S0IES4 Lerantse E-- a a e -- -- m e-- Cte fxm ---- -- Br Neet:ThC1C r esve cidine 41dr md ) cin from theC6andC8saodandcurves. om! = rettr LoLm lls fA = sn C2 LE 1rVtilme,]coeeiLm =aly [rv ses 000395 _--_-- ES LWyinlddlliyfee Iinntteerrnnaattiioonnaall,, Ldtdd.. WW TProojkecetNNuembbeers45v4-e10s5 Dict and SupApplpeemnednitxFormulations Tale 1 `Wildlife International, Ltd. Game Bird Ration" INGREDIENTS hSFoieyeBaeCoavmniMdMeseas,46% Prin i vPreoyn SBpaesceia2l,06P%oriinn DrGiredaWnhLeyips EVMuieabariinonaiCnnadlrPMehinonrien +Piriads (sebelow) TSGoiLullFoedranadFermoy PERCEN04T) pu65rn0 "65o00o0 2aic5so% 000323s Toaion oVpiepni 1hned MlionweingParims,owhuenndtdd 0.32% ofeon, AmocoSeppe PsTo NVViiueentins, 27000000000001S10.a NVFoaionnbienigcs Acd AWomgasn [= BPiFoomcorheide sho|Tmmueems TaVViiamaminmngK (Menadions Dimetra Bis) non5fi3rgradn ZMianneganese Comer 102 apmaem `Hgams Sjonienn PrTog Rp "The amgerr.adsmalyss s minimof 27 prin, mii of 2.54cade and ima of $6 * Fermentation By-Products (SourceofUnidentiGfriowetdh Factors). 60037 _-- 39. RF WL ildliife sIntterniaticonsal,eLtod. i ProjielctiNvumiboelri45n4d-10J5 DiAeppPernpdeirxsion JeP19e,2n00i0,sNfoomriPnFaOlSpwrepraerpornepwasredaosnflFoewrsu:ey26, 2000;Ape 7, 2000;May12, 2000;snd Cont No premix reed. . Lbppmai: 030845PROS+60997 grain 62pmaic LOT gPROS +6099graion 16pmai: 30839 PFOS + 609g 6ti9on Basaraionwasweighed nt a ardHobartmixingbowl. Approximately 10ofa0ongwas t sWhmeaarrilimenaogrbtnliaternos ngwdaesaf Wrdadkareeneiddnnortgtheor blwceebe rnegudnhd edbaren.o.ysTahTwrheaeetsseHsoifteicsoddunbssohcltoiaednmaceteitewosan.sswewTreweeiimghiehsretedidisounanbtpsoprtraanootcxaemrmhwceealdwsymeaiiongnhsgebmobfaomtwlactnwhdaitedha hcroeanntbseonowtmlse,dwweirltmsheiexmeridixnisenpgbpbeoriwnolxuTtmheyee15admhieemtwbeao.sw.iTnhTedopbhroiweel nwwssasawwcietidhghsoendoHiommboae1r0t0sm00ia8rohnaonfosdm Paced napropristly abled psi bogs. rewghe, and stored Fon As ended, the appropri ris ws incorporated int se foal dit s follows: Oppmais Z37Skgrai+o1n250kg limestone L8ppmaic 1000 Premix +2275kgration + 12kgi5est0one 62pmais 000gPremix + 275 kg on+ 1250 kg meson 6ppmaic 10008 Promise 2275kgraion + 250k imesione he dies wee mixedforsppoximatly 20 mines inaPatersonKely Twin Sel leer. 60393 m--_M--Am--mARMAmPPmAl--lA--AA--A--A--AA--TM"m-------- 0 JWHildAlife InOteUrnRatiAonNal),H Ltd.ee iProjNect Nwumbeir 45s4-10s5 AppendixIv The Analysis of PFOS in Avia Die | | | | 6n0399 | _---ws --e a Wildlife International, Ltd. Project Number 454105 Appendix V ANALYTICAL METHODS AND RESULTS TypicalLC/MSOperationalParameters TABLE INSTRUMENT: ANALYTICAL COLUMN: oveNTEMIEATURE. stop ve: FLOW RATE PHmeeoPwkaelirieokntriEtin-nEmPgiacmsmkeoTardruSedbMC(loIoGdEnIeXSl.pA1rP1La0y01sH0oi0g1hsoPMrersaf.sorSOmpapneeccnereLodimqesuirdCqhluirpopmedoanwtoignrwaipthh Keystone Beasil coum ($0 mm 2 mn LD, um pri ie) 30C 500mimes 0220 miminue "MOBILE PHASE: 72.0% Methanol :28.0% NANOpure Watercontaining0.1%FormicAcid INECTIONVOLUME: 5040. REpTroEsNTIONTIVE Avproim3.a6miilnys REITNETENRTNIAOLNSTTIANNEDARD Apprsi2n6smieneys PVrOoNsITOREDMASS: ~~ 498.6ama i MIONNTEIRTNOARLEDSTMAASNSD:ARD 4267smn @a0a0n _-- a. Wildlife International, Ltd. Project Number 454-105 Appendix IV TABLE2 CALCULATIONS The concnirionof POS found te instrament was determined usin the following equation: PFOS (mg a;1) st nstrament = pestkoo.sat(vrinteerneep)c_otenalstandardcone. (mg 81..1) DeterofmSaimplneReasidtuesi(PoFOnS) The concentration, xpress as ppm.fo each sample was determined sig he following equation: PROS mayneBEOS(mg 01/11s)at se.xcextract iadvool(uL)meditidoilotn nfcfactor DeteorfminiitofnQuatiattiion o1.0n0) "The method LOG, expressed a pm. was determined sing he Following patio: LOQ (pm 1 loweststandardconcentration (om 1) overalldilationfactorofmax lnk "overtion croft-Sabin (2 dinre Fortification Recoveries "The ppm .. messured in each samp is dividedb the nominal concentrationofeach sample (fortified evel, ppm .). This ati mes 10 isthepercent recovery ofthe method t that eel of forification. Recover=y_Bom aT. moearsutredeindsample 100 (OAL _-- : | ] Ii] 8 3 i kg 2 g g |i i 1i! i `38 33 2s ssp | 13 IR | i"i . iI |ii g| | 5837 Imp Zmsg Issn ff i: | 1 1 | i i il as B| 322g ses seeg see is LEH ron omvem mniH - | | (of |] 2coco mr nrarsre a mmmcoco dE %EE OEg 1i Li 3 i BLL old ai gd | # 3EiE gu gail chp bd 5 7 Faas Ra fe Ra (lal g 232 333 33: = ik 52 1G2Z]| 32338925 a32393 aE 25%9 aE Tess ii Lil 2 Caan 2| i i i i =3 g 5 g i: i ; hE i ie ie Xi Beg iit ii Ege i Tai|T i i RHHIE gHoRnnCst qHaIsRtsCs pHuHpsEH % sit S 4 [HE 3 id nTmene mEmanE SERasR HH 3 - iili 3 Hi sl 2 i = v 2 i fe: GROADT _--,,-- } . : i i 3 i i } it co gz El EE i 358 i : i 5. i | 2 :ow HH8AEEES - ; | k | < :: S i5l. B i - eee --- wew - vee j;:zi184 Pol d EH| 5 i -n 33 2 3%% 3 ses sos gilt 3 1 t 2 JiIH] . GROADL CL | 1 EY = || Ji 2 a = |: | | f| =5 zee sam ae i mi ) ii | Wag = i? | 2 8 523 $39 8 EZ = 18 Co i Bi | | ff | i3 | 312 go. fHhl | ;sl i6:2 1 1o Hi | he HH :: i3i3. . | 2 li - PC aSgl | | nO: _----m a. YWilHdlife Inniteerrnnaatnioonnaal,,tLtdd. PPorojeecNt eNubmbwere4s54e-1d03s AppendixIV METHOD OUTLINEFORTHEANALYSISOFPFINOAVISAN DIET Pear ai oifcasaimpn ihedidavianfedsock win heymis che, Weigh10gampleofhe maeie.bFrken,cmhsui.efpiascb+ooniss.dResctosrdaliesgh: weighbts de Fo cachsale,mer100ml.ofmea R vithoegraduated yd nd eter keFrc re : Caapndoaonhteresl.rAlelowmey Ssol hefor ii of30im e ' Vac lrwithaloe hepepe ndrieresindfd oes wilhmetal lite Trhe lten20o0m. 1 ver ak and ing vee ih meh. . SOcEhNPoruA.epUNppamptOadtiooarnsnteortonomoeisdsininnfigtlioc0ne,n.ieomn0qgiHou1PF0OFSoh0rIecL (faomleba rrd rangieoh a foboSet0L04dmMs pSaolny Tonsoni 4 Ample dubssupe or LOMS li, Fre1. Anyi method low cha ores ofFOS insindict. ROADS -- -WiWillddlliiffee IInntteerrnnaattiioonnaall,,LLttdd.. ~~ PePrioejetcvt uNmumbbeerra4s5e4t-1o0s5 AppendiIvx 120 ol os | jo 0 0 | | : L0mR006 012 a 018 02a4-- 030 036 042 08 | Concentration (Rati) i Figs2. TypicalcalibcurrvaetfoiPoROnS,Slope = 248673; Inereep = 0.07396; = 09955. Curve isweighted (1/x). nR0an? --_--_-- 8 }m---- i. BS WoilbdlsiifeoInntsernationI al,SLotdo. . Projc ect Numberl 454-105 AppendiIxV intensity: 20000 cps 10 9 8 7 6 5 4 3 2 212 1 28 8103131 178 266 40169 81136 1220147162711 230318 244016 284173STeiamne Figure 3. Typical ion chromatogram ofa low-level PFOS standard, 0.000351 mg aiL. 600403 _-- "so. YWViillddllijfeeIInntteerrnnaattiioonnaall,,LLttdd.. ~~ AppendixIv pPerojcectt NNuummbberra4s54t-r1o05s intensity: 20000 cps 101 9 214 8 7 6 5 4 3 182 2 1 7 2 144 260 0 41 81 121 161 201 241 281 Scan 069 1.36 204 271 338 406 4.73Time Fur 4. Typical ion chromaotfoaiggrheavml PROS standard, 000439mg a1. | 0ROAnG --_-- si. Wildlife International, Ltd. ProjectNumber 454-105 AppendiIxV intensity: 40000 cps 10 9 8 7 6 5 4 3 i 2 42 t 76 198131 150 228 253 540 o 40819781136 122014 126711 230318 244016 246713STemaen Figure5. aTpypiponrcihroertocemxaaiotainogtrmliammaoofftPaFemOSa.rx blak sample (454-105-MAB-1). The aow indicates Goats -- -_-- mrr - e r 52. r Wilde lifenInntaertniateionnaaln,LLatkd., PrOojemctNNuembmera45s4.1i05s AppendiVx intensity: 20000 cps 10 9 8 7 6 5 218 4 3 2 1 34 183 o 88 105128 160 264 41 781 121 161 201 241 281 Sean 069 136 204 271 338 406 473Time Figure6. Typicaoln chromatogorafma matrix fortification sample(454-105MAS,220ppm a... 0noait _-- i. Wildlife International, Ltd. Project Number 454-105 AppendiIxV intensity: 40000cps 10 9 8 7 s 5 4 3 2 1 0 4 7290 _ 128154 180 228 265 40187 g1136 122014 126711 230318 244016 248713STeiamne Figure. Typical ion chromatogra ofa avian dit sample onDay 0 (S454-1053, 1.8 ppma... 00a _-- 8M -- ____-- 51. LWioldbliefe Inteerrnnaatpioonnadl,,fLntdd.. | Pro ojectNe umbers454-105 DiagrAampopfeTnedstvLayout : = pp tm ger . ==FSotuerpXa1n.PCrhroomcanc0.gMotdbeuNso..555 002g _-- [---- 5] ruesa|an In| gazas as Iz FREE ET I=| szpsz|ze Iz I=5ea ss FH] EEEEHIEE =| szsgs |g is 28g~~|=2a I=] gzgag ze bz TgR=v [23 2 FR8RR% | ax 2 2873s | 8e i i=" 2383 |8= 3|z-| seEgs Es HER FEE RR [| FN PPA pe : fed 3 Is ge2ns= [on 35 ky EB Jz] ansoe os 3k FI833 | 3a i Ee sseng| as i be aver |v be 3827| "a i Ix nogrg [ox 3o| sreas |e| Js] =ane se ii.|o] gs5sgzzzzals8e0 81 58838 is Ix saszs es | [1] sness|ee |} Jo essnnss [3 fiolo| gzEnEzszs|z|e8s ii 5| 55838 is 2 roati _---- so Project Number 454-105 ks frase Is Lise| |p| i JEgl] gi d[1| =sssgs|ax | sumea)en| Qf] Bi5i02l|e5| c$oen8e7 o9na | ii1a] 13 7|ee[ samme ee| Sf) EE Ju =un=e 2 af Io gzgss |e %-| 5] zrzaz|ge &| gages i meee |] @;zvasgsegn|e[a=s 1||} wnovveanfeasn|[2 mrss se [1 ge=as a2 i ezpuz |g i sazaz|gs |: Goo; -_-- -- Is $5393| 8% Jo] eves az i tol sszzz|ze| oo] msens|es |] I sl =9zsv|ae 3] ~zase (se |] ssilt File]$ EsEeeEEzE|2E i i=F s282g3888318(g2s i Ba yr sans fon| 3 hl wenee| ne |g FLA ER LEELTERR I FR HEHE SR i Jaf s5mnn|an fof guess i i3o] gEe55s8| 8E"F 8] azzzz|ge ifo)]s823g 32 ii HIEEEERIER |: 6RO5LE in 5] szzas an ia] gegsn|gz $3] sven as Jil amwan os sro 5] sass as $=] szces se 13] wazes es Jz eons ee 3 jz] 3g o|3-| fii : Is seen s Is gEzzg (En| ,|I-| Rogen og ir By, 30710 bs ngoae| ex His rma ve| Cf] go3es [80 be iol zzzze|g I-| I ove |v bh 1-| essez|ee| [3] I $2828 5% ba -| gazes |e fof HELL is ] a2ge" [55 sazen ae EEEES EL IN soya| 5s : nae | gevss [ze i zepez|ae |} sees =a |} svnes|en| 355e8| R3 ! gazaanias |: 85838 i2 ! 004? -- Project Number454-105 =| sazzaes =| szasg| es 2) 2z523 | 8% SEEITIES SIFTEITIET =| 23383 | 82 i ul mezse| es 2 lz] szeze| ae E41 EY azasz|a- iy F] |] Brean an BOC] eens LE] ~| sezza| se z | gzesz| ze SETITIRE | zzzze] en | ezaza| sn "| amass| as -| zazss| ss g| sazzz Is nats "0 Pojct Number 45-105 3 3i3s f5 ii; Iii SE 3 "| ssegg| Es NELEEEAEE 1 | PEELE E= l) zzesz] as "| sees| ss "| sczunl| se 2Z=ay | 2% E| szzas fs 600359 A ----------------EE -------- -------- -------- -- a Projst Number 45-105 5 is ig: !i if ; ol gsgaz |e ol gzzse| ze MEI EHIEE & gEsng| 2' "| nessg|& -| smzzs| es g sz3z38 is (0470 --_--mm-------- a PietNamba 54.105 3 2 ft 2:88 i i z | zresa| as =| eages|es =| seaza|as =| gs282 3% HEEFT =| zanas|sas =| seg28| ze =| sgans| se =| sgses| ge 1 |e merase | ssses| ae ol g2885| as <| anges| as NEEEEEIEL MEE EHIEE | gazes| se "| sgs8g | 8% gl szz28 ie G0042% 61. Project Number 454-105 AppendixVIII Table1 IndividualGrossPathologicalObservations from a Mallard Pilot Reproduction Study with PFOS. Birds ButhataTenstTierzmineatidon --_-- Control (0-- ppm ai) ---- _--_-- Males Molingflightfeathers Bumble foot: Testes small -~20-30cm Testessmall-~ 3.5om --_--_N--otremarkable -- 0 202 P2e03w 204 205 x x. x Sox ox x - x. Sox o - xL.x ++x _--_--m _--_-- Femsies EEE PE ens E Mingfightfeathers XxX - o- ox Bumblefoot Xx - ox x Regressedovary x xxx Eggremoants inthe reproductivetract x _--_N--ot remarkable x- - | GOOAZ "6h ProjectNumber 454-105 AppeTnadbilxeV2 II omIandMiavlilduaarld GPriotssRPeaptrhoodluocgticiaolnOSbsteyrvawtiitohnPsFOS. BirdsEuthataTenstTiermzinaetidon Maks BNuomtbrleemafroaxble 18ppmai. EEpenE s -. . . x xxxx - Females BRougmrbelseiafgotvary Eggyolkperonins Notremarable pens We Wwe 29 20 xSoxx.ool- ox SL x Sx - 0N042E 65+ ProjectNumber 454-105 AppendixVII Table3 IndividualGross PathologicalObservations fromaBMiarldlasrEd Puilott RehparoadTuencsttiiToenrzSmtiunedaytidwointh PFOS Males BNuomtbrleemfrooatble 62ppmai. mm paers ae as xSo-x.ox.ox x- Femles FBeuamtbhleer ofsostod brsionscnc EExgtegnsyiovkepoeg ryoonlikpserioits INooatbredmoamritanbllecss. J-- EWN m am aw as xSox x .ox xc.c x-ox+ S Sooxx. . GhOatL _--_--nmmmm-- "6- ProjctNumber 456-105 AppeTnadbilxedVIII frI aMoallmard Pilot Reproduction-- Stdy-- with PROS BirdsEuthataTenstTiermzinaetidon --------------------1--6p--pma--i.-------------------- Maes E rE E E cere: BMausmblfeeoodt pacedvaderthe ge xSxxox.ox TTeesstes ssmaalll ~2150c-m20em CSorx orx rox --N--ot--rem--urab--le ----------------S--o ----o ----d------ ---------------------- -- e-- r ---- Femutes ----E-- EpeenE -- s te-- r MGeonleiranciongdhitiosnt-saboorsmmaalllblodyy! x~~x. . oXx - ABiurmsblce olot xSlooo. Tl SiRgephrtsesgegdoyovlakrpy rions Xxx xooox x x ----Not--remar--kable------: ----------. --. ----. --- ----. -- !Denotes asmalbodyframe,butdoesnotindicate 4thincondition. 60045 -------------------------------------- 67 Wildlife International, Ltd. Project Number 454-105 Appeodin IX. Histopathology Report CRO4A50 [---- EPL' EE RE* E MoU S-- E EATE i WITH THE MALLARD mtr EJ E rn onds oaar Vitti 1 Easton, MD 21801 a: enpar? EPL ER ~ -- A ATTA Hers oman woe cs wero nase ts ---- Page + JE---- werommensorsen mas --rer--e o o " C0048 70- Project Number 454-105 PATHOLOGY SUMMARY eoing Pict Ner 105 EPL' WOLa FE WTERMo ATIONAL LT, on PATHOLOGY SumARY suesfLoahmmadiulrtsmsalaeianadxeammianiaomnawarstp(onerspnyorsecotio)nsofisclh erde rPeouatooncoanresiloericcoac rpuostcs soo [ePFOSor)fibonetfeiosc 010 ttwneteeykcsi.ooSmeloelctvsed twaisasrauteeossfceroxm iupnntnreeaatecbduyapgmprnaoxriimccastc(elonypay1e2.t-weTehyke-aolirdcofiofsoptfrioonfg etry expose oat unanncvr a ast a swe por. The [A| di oeprins g~ | [on Forme woe [1" 8poC mao PFOn S | | t5 5 ol | TO s 65p [p5 4 ma |i 6 5)| | 62ppma iPFOS | 5 | 5[4 | 6 | g rMail7la6sppnhmohaeic$Po5nFtOotSnn d|17.5 055.1[ e. re5 s wer| emvee5 tro[ r610wo5 ke | MATEARNDIMEATHLODSS euthanize, and accpisvars patbyehdt amin, 16. 600400 J------ EPL' CA TRE A ---- Slotsarin ors othricd atposal 12 wk of gs ung fcoxaoraWbioinnld1fi0ox%aidnme tgaass,saf,nod rsLaae.mpolSemesiafoanrdhiassntoupsaetahonoloogmiEcxaalptervemalauaarntdsiotnPiwaennrtesoconwleleected Laboratories, Inc. (EPL), where they were embedded in paraffin and made into {ahoeakmoatwtiionx:gylsiWnsarnsdgeooasminid-stt,asineradnsdefcotrioinsnvognwgalrenassyse,mxiacrmrionasacdogpooeynsaKlditdesv.maiTyohseactoeyst5, JusorfvFoarbacsien,coanndSapoheea.g14F m aseT , roesn:aBorne rs epveatlear micandingswar carttadcr af IncidenAce Teasleoss, rMiivteesdcoptpreintg aforrreaossausi caHmiecpothmoaaayc mallard are listed in the Histopathology Incidence Tables. Microscopic changes were graded onetofive depending on severity. Nongradable findings are listed afSsouamplmreaasreryntim(lPa)sadnedostmiTsamsuiaeost,wcigteh yaextsren,swivei8ahugtolotys,tisaeanrderilisrotedaaasni(Agm).rsuAplrl fain1ds.ings tousoarviitahoafugossswceseemeartoansnted stncroptywi ta coforGersopsasnadnidngMiicosoconcs Fshiange,ateps.raTtheta,nteeshaestseaitroCoroerea --. transcribed from the Gross Necropsy records providedbyWildlife I. nO4ie res EPL' i S"E SY rt) iBESnULTSremeE m ----T ------R --roE n iein.tterseettootstiots t iovyotmnspoe ia , tee-- to st-- n cit-- e omoe secanstnots nt tr tom Ga r Sor-- i one-- tmea-- ntve era Ls) a rec--oot--aon--senns--mehte--neo--ms--y--ne--at e--y atne EteomemnttoSmomeesmte do oneee 8emtrns rr ------------a:--s--ar--a ------ao--asi; ------ Feene ten meaty on re, ts rion Avo--tr--e o--s r--o--mnre sts may t bct s et va -- a ------------ CNO4AIT ProjectNomber 50105 EPL' ERPERENTAL PATHOLOGY LABORATORIES, Wtator1,Sty rte 4.08 h(ycslogkmcpliaomumrtaeteutitsy)on. yToiusrmo(rpphoolosgyewtasZcok nswiidtsnsotrtmsharhn a poroOovagriteseoffmanycontro an 17.0 pom 1. ah fees axis vos acctoonracspootonrgdrooaescdrsobesmeertdvere.gDreecseassevdarty.cAfecaimdectelrics vers Pmiabtresrsawnadsacdhasrhaocrttekbsy troawnlniodriotprofomaciuslcoamcnttosvattoem.e ccoantseisrteedntrwiougehahlepeosvtarrieapnrocdoucet.iveTphheasseeorveagrrieasnsifono,isnovremrasl mpyssticlgica cShoennotmenndcn.17.S6 pcoh.e nfaomneceatandrdugrss, nofyhweissstcognssweerredSoeren dtnsOtvtaraienseosfamafnri,ng femaleswore charctrizedby rumarousactos and very sma, nenpadunciato ollie completely ambedd na proment oimeate in young approstly 12week) ics rane han patra `ovarian cortex. This morphology isconsistent with normal physiological adipocyAtesdwioth ssisglioofrgadpsdadnroopsf,lamocngowhniwsfosirhufdtesnasgcetrweiade Taomwoasdoallrsesawhmeonpfcolnptoeisn esliiaex,iostoadlaefdocfrhsasamueste,raWdhoares, iincctroeavseesdi.atWaiisoonvewseiscduaoigornosweatsnoly lhiennod3h8camoabtnoferoeerstleessncalwnards iontcorreansgedmiancnidde,ncenwdewsatsea donyu a1ve1r7yul5t5m,a1m. aidutnmaln.saAmpcareyd contol au males Tis creas may hav boa rutous overt sated ONOAIT 5. EPL' BERNER PATROLOY STRESS js Nan 5.105 teams, Sore08 ibnyempaosrosufiaztsorrpiolsseibflecvacraiotytien seettlhedsosta.plingte, ut ocpointg troeteis ocvdeonsed ress, an wer coradred Incidences of adipose tissue microvesiculation in male and female a0 avnedrcafsaprtiongnmalairccfocmconingaansrtcdrgrobuplso,wnvirnsgnti lmaominnaopmreopraccyls,aendsvainivoerosf pheier:saKoifnFayamriekr.ainzaoteonneewseaens coampnarseedvewnitylveesr shnra.hFarnhrseaiesotomsahsscegnansitwaercgonotrdwhrreed detaWoantonrueetlceolofoatterslin va(rFiuOsSsaimisnactoinosni.d ofcrtfo of Cmoarhof coosowoihrpsycadsnrtrmaacnecemaatnomacophargnes.sSretoretd o oVuitesmtisrocetson.ed snongia0.2kpinmraa.xifneernsaimnlg,misnagcpkroporfe etsetr Onlylined abnyefaullattenmallard (a contol ml ha rsa ed epithelium andfilledwith amorphous,Fac svar pale basophilic material. kon expecaitdisn, Toorak Pan was rset on microscopic evaluation. The bursa in this individual exhibited the age-related unremarkable in all male and female offspring. rLivaernpsiAgddpmireteinsondeanelpfioendriptngsecwoeenrteaoalcmsoaofnaoimtek,deiSn,tthereloiveensrtoef sgtorapcowpemanrten adults and/or offspring. tlhee. The ef identofittheyon st pigment contwas notome ated determined, but ml dopoton sortin may have been related to g Croaid cr---------------------------------------------------- EPL' ERENT PATROLOS ESR oRES WS ProjectNumber454-105 tsrt 5 me16 dA egeersianinocrots cured oy in fon corte a tnt greg. `of amorphous, pale biuish-violet material which filled and distended the hepatic pcarrctsinpfhoi,n asedlncdrasuoersm,iannapgamtartteedsowriceoeelrus Hepstociardognaaionnesesswas carr del and orl nadand tering, etyhang det srysid, shar beedesmiearntceasatehdmi,ecclceaoravtamcau:olpesot(teconSseiasptmeontawcirthh vsfaatosaactcotusmuwalaitneiona)dteienrttrhoeedsutel, eTopcsko"naevpe EeorgsvAerrpeasnnoecniee.' nidhwreenntononn owcetenpadtroosae,batyvciaerggaecsurted8spoortadcay a ownaresand dierra.evrInycnutoatntleanoga. tepatustahnottFvaeulatwe sve adrnicaelsnwt ahycstdoasneddsmagtuemssoifosowvonrsswwareeessomotimmees nbotted seas orshlotesvaryeeaferwir reldo biological variation accentuated by small group sizes rather than being test article effects. CONCLUANSDI SUOMMNARSY tarniisaNntooolrnesbwioonrsrcootfnsFeidiaenrcede,rpo,srsnaibmlyy,reclaruttedmtvoltes,at anrtimcglleu(elPFaaOgSr),dovoar, Jaane - EPL' EXPERMENTAL PATHOLOGY LABORATORIES, NC. Projet Number454.105 ht maton Li. StyNumber45105 fpring in he acipose tissuof acu females an ofring of both sexes, of in ihe tetes of cipring males. posteprToesdtuecsiovfeapchatsemarleegsreesxsitoinb,iansoervmearlalpfhyastiucrogsicmasltphceonnsoimsteennta,wih itnucbluuldeidnigaamestpeorr.misG,roduecprfeeasreedncspeesrmnatthogeeenxetseinst,ofantdheosrodfeicnrdienagssewdesreomienvifdenrto,us twhielhdeagsrpeeermofidoecccrueraisneginoslemiinntihfeerhoiugshtduolsee(d17i.6ampeptemr bao.in)gagdnulemraallesm,oarned phreosneoufnicndeidnig a1r7e.6supgogemsta.i.veadofulmamraeleascktahannceidn ctosntticoullaarurtegmraelsseisonOvetrhael, 17.6ppma. adult malescomptaocroensd, Adu males the17. ppma. Gicrroouvpesaliscousetxioint.edThcelesaerltyiicnuclreaarseadndinacdiidpeonsceessofuasdifpionsdeintgissnsue17.6 ppma. asidzuelst,mbautlstemapyoshsavvietybeteantftohretyuiwtoeurseerveelantsedatcocetnesttuartiecdebyadtmhienissmtarlaltigornoup cannot be eniiely red ou. malarThfeofmewcoontthroelr acnhdanrgeessteidn vgarroiuopuss wtiesrseuecsoonfsaiddurletdanidnlcoirdeonftf!sparinndg - unrelated to test article (PFOS) administration. Ge Veterinary Pathologist \/aslor 7 G6o0als Prjct Number 54.105 EPL' TTR CUAITYASSAULCERRTAEIN StyThe: PE0S: APURaSthywo i nala tansy. 454105 prConiaer Magara Grist EPLProtons 22025 EPL Papo Daria. Grist o a To loin apt caotfeisSascywrveeeoebpy gacsopioGcsuuvtyeAsessesotrcneiUonkts| -- en ones = Erte rons, une nnn Pops ozone sar ees Woiroms one swinwien | ares N- ono raseamn -- i pune son Sects rn wo el LZ lg t ee ofzefor err NOAY _---- ee ---- 79. Project Number 454-105 SUMMARY INCIDENCE TABLES ADULT SACRIFICE Gnosis --SE ------------------------------------------------------------------ AWiadslvtee10M8aslalcaertdttce _s0SUMMARY INCIDENCE TABLE ProjectNumber454-105 plete=f |lafiiEate, MonomicloarGott| 1 F-- Eor --Be-- oso-- --)------1-- E-- E ------ PBRoSemoF VRE Ge. CC------ mp---------- -- FRLLBLAT G0. Soom) -- |--|-- e --E---- -- i Spere mfr r] [[limn1 eeslsTlerlaetaei,loWonnomuclomr Gott| Jp [CmFrIeVTEroGo todbs B opoatmeE len ------[--m --e ------------------------) [FPea poscoicyieace), Facey Ghmmge,|_| [FC FeopaetE ocsyeeeetCarOrP,w kFacTeyEeCh7a2nr gCs,| oeee| | ----ss----FF --r] ---- --) P(r eEe dsmeter 51 y brp oveveermr mo] [Eiarfsifreerreacoeo,r--maessnsmactmtien--nsecteorrE ] e --3--} 1 e ] I [[PFFheyegDmeeicmmritofseenr--nodm--Btlo--absu--stt--eesr,------| 1p [[ 1 5 s 1 1 ] [Sr r peematopanote, beeromsed [5{3r r [1] r r ---- t r r r . r r rr r r rr r ] 1 : Err ExpePrrtogneobaeriona,1lc | RQ4Ang cmrwilinvaes ---- 2 e [Mr cros vesfeul[ att 2 iIon|r p r ] BRAT 00. BED) me ERTFTREE peeieee Jr 1] frre reefeee} [E1 SENED ] {Tm{ vEedSebCteOnsRTpra 1 r1 fl 1 ) FALLBLADDER(ho.EXNINED) || {nfilerate, MonomscloarGall {5[ | 1] 1 ----1----1 RTTOEDT IONE 0. EXWINED) jog emoreremm foememe]eee] | 3) |; | i = b [Mnveralof--asaotioomn J 1 FIER o.BRED (Amyloid Deposition -- |v my 1 11|--f------f11 [Hepstocytes),Fatey Changs, | fie {v3 | | {---- fp----p11 F= og mmm = | Fae EE TR ---- 1 rrr RYO.BmeD [7 ------------ [|] SSSS S--SA SS R-- [Follicles,Decreased Dlamster | & TEESETETEI drt |5 | | for me fee ----|----1 | eee meyer ttt] FROVENTRICULUS(RO.BRMINED) | 5) [InfTiteate,Wononiclear Celt[5 |(|[|] | [| ----]---- i F c ------ l e-- eSm-- eef-- e]r e -- f -- el] eee1 ) or ---------- r ee rer pee --t----1 i --_-- rt ee me ee e e ee fe-------------- frm meee 1] eeeJere] 1 e-- r-- efr ee ---- ----te e -- eeet SE 60040 82 Project Number454-105 HISTOPATHOLOGY INCIDENCE TABLES ADULT SACRIFICE -------------------------------------------------------------------- 8 Project Number 454-105 A4d34i-e208Sacrifice Male Mallard HISTOPATHOLOGY INCIDENCE TABLE cocnrrhooLp. r1o.v6e 3 C PrP ee TCv ar x TTT ET eCS-- 3EE A || E -- OE ---- w I --S ---- eY w e --A-- b[eC BCSeArorFmPagaRTeeos -- | {wwT[wT(wTl[TwT{w[PwTlO wT wTT T) ] CEVETEnfilee--atS s,--Hon--o-- ns-- cloa-- -- r Gol]-- [3[2[x0a 8 1 [3033 ] [x0 1A11 EE A -------- ------ S-- SSS YAE-- 3CF Y7 -- o| [[iPinmetc{aTteriaotas,tiFoonmomelT enr Gert TT T[[TatC { [af T 11 I [EEer peesocyitece),Facey-- Grange, r |0 [|[T A r TT E-- [T Heps -- Factey o hangcs, y-- |t {|a1te-- ||yt , n { -- A 1 -- E 7CT-- t -- eYYe 3 Fteea | fEoETm Y yu 3TSSCe--a VM EW EWEW 0 [ EJ EI3 || Cr EC [EI TSCrea VI SEWSEWEWEW0O 3 E010 Ielelelwsl fefslareral TTT111 I [_DF ecressedD-- io emete-- r --e J st ttefe2 tstsX a tai l [FSpemstogenesis,decreased {r4{rz|[rTrorl r[rlrerlrTrT r Tr TTr T] r --r -- r rrr rrr rrer r -- rr rrrr rrrrrree r AA =| pL r En m---- a ms te mis sant Fm BR a, ey Son noisoRmE | _--_--m---- Aasdiv-e108Sacet ice Femsle Mallard a. ProjectNumber454-105 HISTOPATHOLOGY INCIDENCE TABLE caornotuern c1rsoup : ---- ee C E E2 Te 3ro SC -- S 86 0 33 31 PE ------------------ wes e A eA rr 0 [PC URERCoF7Fasc -- Iwww ww] [wwawO n 1 ES tS FE ET A 30 5 31 I o [Infy iTe-- race, WomowetousGol |1 Tala 3 n TearTf T EC S e-- t O A a C 2 YC C T2322 -- -- O O [rS eperTCPoFr aTttyC dhr,t A TT S ES C------T ------E --YYENESEH W E6 E3 133 Cb -- C L 7 -- -- 3 r -- 1O -- [Foi llC icles,Ducreus-- edDiameter[3O J3TsTs{A 3TalEsYisis TTTTTT11] |i i C i Fr-- Er O CX C 0 4 E 0 | ||! 5 A AA A A A | EB 7 St A {! 5 A Er a rs HuyEtaE,Rvi mts SS mn G04 Z EE -- -8s- Project Number 454-105 CORRELATIOFN GROSS AND MICROSCOPIC FINDINGS ADULT SACRIFICE 604d -------- {lala Jaa bi Ln] Hl Po, i sifd o[ l! 1 11 0 [IT4T 6004 - 8. Project Number 454-105 SE EE J HYRE Yh | al F! L: i 8 iy i : EEE FHE 4 i J (i 04d[14 BIT] Nos G ! - i Hod | Lar bi | ; Il; 40s : ill]11]] | Pl, 25s a2 8a3 | oy Es | g 3d]WITT eros SE E : [iR i g it : Biff ya [LLTTT G04 5 _--_-- 90. Project Number 454-105 SUMMARY INCIDENCE TABLES OFFSPRING SACRIFICE 6N049 _-- rOifsfaeperaisn0g8 tsacrifice Lo. SUMMARY INCIDENCE TABLE Project Number 454-105 [oP rmemi os oxme |rEo a [o [ POIFCEETISSUE(RO.BUHINED) | (5) | 0) |b | 55 |] str mper [M o cr ro m v~e~sf | c3ulation 1 [Indieeste, PRATN (Wo.BRED) | 57 | | wm 1] or] WonomToms Gert [~~ Iror SURBA_OF mE FABRICIUS me---- (WO. tF oo ny t--otot tt o t+ (ENED ey CHEETreader mw] tee] meee emir] FALLRCO0ER(io.EXRIREDY | (5 Jnfilccace, WononuclearGIT[5 | |m 31m 2 { |o 7 r ----f ] --| OTTERE EEE Go.EONINEDY ~~ ertem yrpe | (5 |mw =m] mr] [nfiTerats,Wononsclosecolt| | verso -- 1 2--{--------------| --t----1----------) HE do.BRD [oy mw emer -- we] | oe] [[Epeoplaetmoecnyetreast,oPnaNtetcyrCoahtasn,gsF,oca| t| [ | 31 1[1-- [-- 3 || 1 ----------} [phe-- --_ ---- tp] Es eS | fgFleopsetsoclyees,Vecwollmatten, T || T ----------------| Ft [Iafiierate, Wom nomm uclr oarGe ott[ t5 t[4[t 3 {a t --t ---- {Pigment Deposition bere Tear |-- 1 -- 13 -- --]----"b-- 11] [mfilecate, Wononscleas EOTE TE BEE cell |5 | dreef 4e| ef &en 5 e |--r --1f-- re--ee]] ett [masses [afilerets, -- Wonomuelons Gott[ 12 51|% 217 1 3 5--]----------]-------- | [r-- e --e --s -- ei 11r 1 e |1 Je 1 r 1 ] r ee fm t peefreeee] Ie fe -------- eeete ft ---------- eee meer] | re eee ef reer) 1 ttt ttt rrr es ery `ExperimentalPathologyLaboratories, Inc. CeOES0 OF4fe8fm4ua-pl1re0i8HngallSaccraitice a. `SUMMARYINCIDENCETABLE Project Number454-105 Em TT] Tr Tre optiI 1-------- --| E r [HimceEovmestcielne atm loy nt w Le me m t -- 1 [fafrolteaTem hmstEsee,TMoEm no--nue c-- lear GT oId T_m |e ------T --ee e ---- w --e-- }|---- f e ------m --]-- --l --------)-- [Ibna fieleeeater,Vosnr onsctensGott1m 215o 5 r 7--]--] --1 K[iEonvewieaTrtemwsootsrB ,eWony onueTarGT[m 3[6 |o ----[--5--r 1 ------F-- [pFF IoHpoaperGotcoorc.ytsteess,,epucyee y Changs, |m b J 1w r w e m r 1r ----] 1---- F[[FFEeepr peeteoocciyyeeecrrt,,VwFs aamtaoilyetmcmhaamr tntgomen,i,--| a --1 t -- t 1 f T ----} ------------ -- E b[FPooRtT gisreinertepseopsoFrw-- sotmtoomn-- etoms--con-- t |f p Re y Si p S--"s -- 1 --------] ---- [EFb nfeilea este,Wonomwo e-- lemrGo| 1 a 3 e r r3 e s 1 [[IbEnrfTteletGeaoteE. Fom nomete-- neco--ri-- | |---- 5 15 Tm 5------1--| | [-- -- Fosaten espels,oper-- t pt r rt t--tt --tvt | r r -- r rr r f r r ------ r p r ] p Eel Expert 2 FebogyLaboratoTrni. 600451 _--_-- 9. Project Number 454-105 HISTOPATHOLOGY INCIDENCE TABLES OFFSPRING SACRIFICE. GOAT Tcaisfiuep0esinag sacitice 55. Project Number454.105 HISTOPAcTocHnrOtoiLnOGY INCIDEaNiCoETABLE apoesr : [oTPGSE IIS {Infiltrats Monomiclear Goll | XxX[xXx xx xxx TT || | [ [ |[TT TT +111111] [Hcrovesiculation fires 3 TTTTTTTTT] bateled litt TT pm | Infiltrate, VononuetoarGet area xa ax Exe |[|| I TlTT TTT111 pfrenerB rE Eg FaredeAtt a rT {iy e S TRIe rrrrrI 3r SrY rr S err Y rrr 7. ITH [bienten Fememeeteas GoI rr{T 3 3T ] or prDr TTF rIO rr fey [InfiTerate, MononuclearCorl al (Gala][alalals]Jalsa TT] (313 [ala [aia fatalTT1111] fr-- A eee ------ r-- r -- C rC r-- TH fRepstocytes -- reine epee TT TT Feet{I [[b HT ei pat toe cys tes Favey hangs. T || |IT |T 1T 111T 11T 11] [--Hepatocyres, Fate Changs, peel | | I [TTTTTT rT 11111 Er TH [FEepatocytes, Vaewlteatton, (oie | | TTTTTTTT 11 TTTTT Tal TrraiEl {alata {slate T1111 fIfilteate,Vonemetorcotl {Pigment Deposition (ial aa paar 11 TTTT O T T] TERRE [FECPHIRINNE. eetds ofr eeereermere-- fede fede ef tT fo l=Y=fy--{fnfoerjmy TTTTTTTT 111 {Infiltrate,WonemcloarGli |[313131 |[alifajl [afafafalTT] Es A eee TT A TTTr fImstute ---- [Infilerats,MononuclearGoll Te][[[*] [elelele] [FIF[EIE || I |] {13 {| | [alJz T I TT 11111 ---------- FFARR 1 Tr rT fe t -- aA ed -- e ft-- TTT| r ------ r ---- rr r rr rr r -- rr rrr-- rrr 1 rrrrrr rrrrrr A er I. ElFgLr BJ SRRI EE EER , SH EoeEs en enOasz EE -- onsets HISTOPATHOLOGY INCIDENCE TABLE Hale Mallard 3 pe Infiltrate, Mononuclear Cell IA O TH acai CCTUS-- ------ J EY I 0 rrr rH| | ns TTeease, Wononucteoar ett 1[1 3 | 0A | T1111 PSA or FARRICIs eS {x x (x x A TH rH mca oh se IE a fInfiTerts, FomoncTosr Gell _[YafYala3Ca3 0 0 [1111+ fe E e S rrr rrA r r A H TT-- [(Hpespaetsoecnyetreast,iornaWceecyrohsainsg,sf,ocal|{[|3[ 1[| | T1 A 1A 11 tm TTSS [Eepstecytan,Fatty hinge, | |[1 3 I [T 0 TT tm reel [Hepatocytes Pitas Vacuolisation, TT || | | C rr rr T T T T [Iatilerate, Fenonuclons Gott labs [11a]J {3 | TT H 1H P ee e A= p ERPREe BDe I= } ITT EC Ta MWEW tt mm pre mee rrrrr0 rrr r A H elses TH |afieents, WeonetearGott {| r Taar l[r Tr T rr T r1 r 1H ee iRB AA 0 00 A eH EE A wn ---- ExperimentalFaihalog Laboratories,ioc FElI SS - hoist Oafsfit-u2p0e8ing sucettice Fenaia Haine -%6- Project Number 454-105 HISTOPATHOLOGY INCIDENCE TABLE conantoruor arioonur coorme : Es A EOSETISSE [R[x xx xxl| Tx Tx JET Infilerate, WoponscTear cert || TTTTT 7 TF 30TTT [1111 [Pe erovesteulation ---- r T ietT uteimeiE eteT te odCdafa T1 T] ] pram RIX (xXx xXx Ix x xx X[x]] {Infiltrate,Wononeciearceri | | | [ [TTT 111] SS -------------- SL SCS NE OTTT [BURSA OFFABRICIUS CPEB IX[X[[X|X| [IXIX[XIXIX[ [X[X[X[X[X[%]] = Loto Yow ime fotoFmTerr poe ee pdt [ICS nLIiEleNrTa)te, Wononuciearcert |[31||N | [aja] [S ala faS jaji] Jalil] feo ------------------ eaeta atera tee tele terete roIonfmiyltrate, Wononuclear Cell Sae m|Walnl (B slelalslalal[alaElYsElYaElEeNlEaNl] J pra J T r y SPr r =e rLrT T== r d T --jyT i=tT =] BCT PR DegenerationNecross, Focar|| |Fepstocytes,Patty Change, | | | | | [a] | 1 I | | T1111] | [ITTT [[EsT ppaitotcyatess, Fatty Changs, || [| | r | TI [TI TT T T ] ] {--Hepstocytes,Vacwoltantion, St a eBI || [| | | TTTTT T11 I TS [nfiltrate, Mononuclear Goll ENEYE EY EY BN EEAE EY [1(3(a(i]| Diiiiap Baal] [Pe viaggmyentDepoe sition --e TT |e " TTTe TTF -- T Tr ar -- t t11e 1 ES ETETTT et fwIz eE tlwl AElAEL edEe EEEAE {17 [nfilecats WonomscTomreett || | | [afTTTT TT OFT1717 [errsERT reece it fede lrorlo Sry eedeederode LLC3S | Inffitcate, MononuclearColi A 3 3SO A9 | [3 (3(33 | JT {afaa{a| fafa] [afs{al] fpg r rerer e seer ms Jee ti toentombmdefr s orrr Srer eTr et r e e rrT=1 [LemingPropets, Geet 1[I [| I 1 TT[TFT TT] | rr rrr rr rr C r C ari ErEma To vs EE ERETE, Uwm,E SH ,nie GOOARE Penale Mallard A HISTOPATHOLOGY INCIDENCE TABLE : forgeTRE --Twpxx TT CTT fInfiltcate, MononuclearGott || | | | | | [TT T1111 CTT ECW A ------------N, SE Sa WCWC [WA NO WE WC 0 So' L73 3NO 3 |Infiltrace,Mononwclearcoi || [ | [| eE rrr mmp~ Teref~ tm ---- T tre [POREAOFFaBRrcTos Ix[xTwlxixl ITTT TTT FESR" t TL TAT tod dfmeefmf meal} (GALLBLADDER -- |Tals x(x][TT |Infiltrate, MononuclearColl [31 |IT TTT TT111 LE A seemedre} sweat itisTai deaf Efe enfed] YY" 7 I i| |Infilteate, WononucToarCot [aJaf 131TT TTTTT TTT111 rcp yt f 1TLTT [D CEe E mTm REmmmmereeyorlrlfergre ireler feiteme iomdYermeei n}e [NITYTTT PO SN 0 J| |--_Degeneration/Necroats, Focal| | | | | | TT 1 1 I | 1 TTI [Hspstocytes,Fatty Change. [1 I TTTTTUTrT [opitfuee_-- -- TTT] | Hepatocytes,FattyChange, | | | I [TT[1 TT TT 111 gE metre TTT PEE TT ---- Infiltrate, Mononuclear Cell 111313 alTT TT TTTT1171 PigmentDeposition 1TT TT T T TT CEEwy nifum TT 0 A | [Infiltrats, Wonomseloar cori || | [TT ecEce cEEeereseeejodfrle eer tei l teee trlie sdrfti oere ird PROIVnEfRilTtRrIaGtHeE,RE.M--onroneuecleeasreCrelsl -- daterri 1 TT 111 [alalafafal[T TT TTT TT1171 | NS ------------------------ SS 0 NOS OS 0 OS 9 | Freee HEE EEE i -- PE-- S ------------ tT | ------fe ------------ i " Epc logLabrioec. iSE BpRs eTmOL r------------ | GeOASE -08- Project Number454-105 CORRELATION OF GROSS AND MICROSCOPIC FINDINGS. 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Project Number 454-105 Appendix XIII AdultLiver WPaege5)|firomg aMalh lard PilotReproductionStudywith PFOS _-- Control (0 ppm a.) --_P-- a aLiever FUeLieverm mwm p18e74 310.517508 m 04 n1oe7m4 1196.7045 us 16441 2030 Men 23% PF) 49 120 sam _-- -------- --_-- 18ppmai ren MaLtiveer w6n nuamu 8 5 2n5o8w0 0 um Mean 251353664 ---- EE Fens Liver 3"15510650 5a2s1s8o5 suse 9[801 -- 600375 _-- mm -- -1s- Project Number 454-105 AppePnadgiex?XIII APdiliott RLeipvrerodWucteioni9StugfdryohwmiathMPaRllOaSrd ------t62-- ppma-- i ------ --_P-- n aLiiveer FeLmiavleer mnm n253m04 0260408 2m10 ansamin nmaam ais ase nom Mean a45s00 n"aosa ---------------- -- ---- 176ppmaei se-- e pen vLeiever e FeLE imvaeer _ a2n6 2148835s 2aemm amms w20a.1s08 u17s35s5 20 1336 101s Mean nsaoin m2e5a0s ---------------- 00477 15. 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