Document 6pQvMXQpO6eqpdnxXeooY6e6

eteree see -------------------- Pros AREPRODUCTION STUDY WITH THE MALLARD FINAL REPORT WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER: 454-109 3M LAB REQUEST NO, U2723 FIFRAGuideline 71.4 OECDGuideline 206 AUTHORS: SReaaynPm.oGnal1dl.agVhaetrionen Nn aber t STUDY INITIATION DATE: January 22,2000 STUDY COMPLETION DATE: August 21,2003 Submited to E5iM55 BCnohrpoAravteionne St.Paul Minnesota 55106 Wildlife International, Ltd. E5a5s0t8onCMomamreyracne D2r1i6v0e1 (410) 822-8600 Page 1 of 202 Gon Wildlife International, Ltd. Project Number 454-109 2. GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT SPONSOR: 3M Corporation TITLE: PROS: A ReproStduduywicthttheiMaollanrd WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER: 456-109 STUDY COMPLETION: August 21,2003 ObytOhe vUTSheerBstuoodyorwnasceicenoend(PuErcoNtteVedcAtiMinoCnc/oACmgpHelEnicMayn,c(e9480w)iCt1F7h)R;GoaFonaddr JL1aa6pb0ao,rna1tM7oArAFyuFgP,ursac5tt9i1c9Ne8o3Sh;tSaOandnEaCrNdoDstiPafrsiicnpacutiibpollniessNhoeo.df Seen, hae Producon Butea,10August 1984, with he following excepions: and 3MCCoommpplaertaetosrigonveedmaontdidnactleudderdepiorthseforrepbolrto.odSaunmmtairssyuetaabnlaelsysaers cioncnlduudcetdedasbaypEpxenydgiecnesReasnedarfuclhl Tepors will be submited separately. conducieTnhcnsadtedly ownasesomrcioootnnodssuocwlteerd(eWucinodlneddeuecmtueIndlteumlnadtepirroonttaowlc,oolsaL.didd.iTtihsoetniadlnyisenfpuamrabanteedr p34rM5o4t-oa1cnao0ll9ys)t,i(cEaaxlnydpgoetrnhteiRoenEssxeywagerrecenh study number 023-066 and 023-070). conducteAdnalyssecscoonfacnge,e cwointthenGtsoocdondLuacbotreadtournyderraEcxeygsenStRaensdcradrsc,h asntdudfyenuulmsbeofr t0h2e3s-e07an0alwyesrees rnoet epored separa in compTlhneceanawliythicEalPpAhTasSeCpAesGfLorPme(d40bCyFERxyPgaetn1R9e2s).earch under study number 023.066 was performed STUDYDIRECTOR: Ai ' b Df SSeRnPo.r BCiagtaggsh,erAvian Toxicology BAT3E 00g SPONSOR'S REPRESENTATIVE: e Newited sul a H(2z A odin - WiWlidldlliI iffeenIntt ernaetior nal,nLLtta dd.. t ~~iona Projec' t Numb,er 454F-1e09 a QUALITYASSURANCE STATEMENT bCyhtohe LUahSbi.esrEtsnotvruiydryoPnwrmaaecsnitceaexla(mPiErnoNteVedcVtfIioorCnIcAoCgmHepElnicNayn,(c5e490)wiC1tF7h)RGaopnoaddrJLa1ap6ba0on,r1aM7tAoArFyuFg,Pursa5tc9ti1Nc9e8o9bS;taaOnn,EdaCrNDodtsiPfaiicnpaituibpolenissNheoodf o555e0,aAgfintdinags wPrroedurcteiopnoButetah, S1u0dAyugDuirsetct1o9r84a.dTLhaebodraatteosryofMaalnlaaguedimtesnatnwdeirnespaectlioonwssa:nd the actviTy DATECONDUCTED Tpeesptruibopannce nay22,2000 Mavi orifaion ~~ Jamary 30,2001 BoCcdooywtmeioghtnand Febuary 6.2001 Haringbodyweight May 24,200 frdedaoldbyody May1. 2000 ggrsaerlesirckness Jy s.200 Canting sy,2000 AReDkS dss cry Ags 14815,201 adtyriada snd Oocotnober 20, November 25, BGaitoalnoagai3nad Decembers7,, 10-12,200 Fat report August 20,2005 STUDYDDAITREERCETPOORRTEMDANTAO:GEMENT Jay22,2000 Jay31,200 Janay 25,2000 Jamary 31,2000 Fomary, 2001 May 24,2001 Maya1,2000 Jay, 201 niyo,2000 Nop 15200 Febmary9,2000 May 25.2000 May 31,200 iy 6,200 Jayas, 20m Aug 15,2000 November 5.2001 December 12,2000 ope November, 2001 March 11,2002 Aunt, 2003 eRobrer.PMeGles Quality Assurance Representative hy2n, e2m0a CALs - Wildlife International, Ltd. 4 REPORT APPROVAL Project Number 454-109 SPONSOR: 3M Corporation TITLE: PROS: AReproStduduywcithtthieMaollanrd WILDLIFE INTERNATIONAL, LTD. PROJECTNUMBER: 454-109 3MLAB REQUEST NO. U2723 STUDY DIRECTOR A' {. Lien SSeeanni.orGaBilollaoggihste,rAvian Toxicology Falox BATE MEMISJRY PRINCIPAL INVESTIGATOR: SREAcie,CheVmainsitoyneJn), Pn. MANAGEMENT: BsAhTEals R Director, Avian ToxE icology ini DilrleofcCNhoetnmi,ostrry pe -- ll Ghani - Wildlife International, Ltd. . TABLE OF CONTENTS Project Number 454-109 A Sr rae --m--m--------e} A-- TIS a oT -- Iho2 daevniinicFeaeidonand Nair a reo -------- I--I ---- e CII DD iet Preparatie on cc eeerrCIeI ee----3IH J a ---------- -- -- F OAbdsuervBaotidonysV.icr i oweaonrd vFoorndeeCrreoresnmensovereon --1 AE OU Blood Cao ll. Fg Collection and Storage. -- III CF n0ddc Inlcubantiong... Offspring Blo nd Tie COUSIN Ce1s reesei : 20 EF re Shellm Thickness Measurements and ESg CoAmpoAnents Colecton.... Resultsand Discusion ; CII fy2200 DCMieltioAenarlOytbilscavlacRieosnhuslotse...neneers= eoeI : , I Tn A S"AduElt Necropsy and Live Weimghse..e. rerI eeees ------m FSOpAhOIOEA. rrr Is Reproductive Results over Ie Goan _ Wildlife International, Ltd. Project Number 454-109 "6- TABLE OPFAGCEO2NTENTS EOggfShefllBTpoHdCyiKIWeEinSghtsg... cc I CII OSprringLi OIBvSrEnWVREIgOiNgS.h..srr.r.srorrrore SECII ConclFusieon... SII CII References... CII -- Si2s 20 TABLES AND FIGURES Table 1. MreoamnaMMeaalsluarreddRCeonpcernotrdauticotnsipopnm 1SOY. .1) ofPEinOAviSan Diet nn 30 Table 2. SRuemprmoadruycotifoGnrSotsusdyPawtihtohloPgFicOaSl,ObAsdeurlvtaBtiirodnssFforoumndaDMealaldaErudthanized durthie Tngeter p-- nn) Table 3. MReaenpAdrultoBdSotduuydWcyeiWtigthihtP(opE)nSfr.om.a.Mallard nnn Figure 1. MReaenpAdrultoMSdatluuedBcyowdtiytihWeoighnt 8)PfFiOo.ma.M.allard in 3 Figure 2. MReaenpAdrultoFdeSmtuauldecywBitotdhiyPWoReOingSht 8) faoMilmlard rn 38 Table 4. MReaenpFeredoCodSnstuuumdcpytiitotnih(PoFbOinSnds) from a Mallard a 3 Figure 3. MReaenpFeredoCodnSstuuumdcpywtitiotnihP(oFgbOniSrdday) from a Mallard So 36 Table 5. SRuemprmoadruycotifoGnrSotsusdyPawtihtohloPgiFcOaSl.OAbdseurlvtaBtiirodnssEfurotmhaaniMzaeldlard at Test Tenmintion.... po 3] Table 6. MSteuadnywLiivtehrPWFeOiSgh.t.s. (8) fiaMoalJ lamrd Reproduction -- 8 Table 7. SRuemprmoadruycioofRnepSruosduycwtiitvhePPFeOrSformance froma BMSallard nD CARL Wildlife International, Ltd. Project Number 454-109 ES TAOPFBAGCEOSLNTEE NTS Figure 4.ReMperaonduRceptrioodnucSttiuvdeyPweirtfhoPrmFaOnSce froam Malt lrd n) Table 8. SPerucentmaogefmsR(e0par)odfrurocmtyiavMeaPlefaodrmRaenpcreo,dNucoiromnaSltiuzdeyadswith PEOS : a Figure 5. MTeraonmRaeMpraoldrudctRievperoPdeurcftoiromnancre,dNwoitrh PmFOasSslPeisczeneagdes (4), A? "Table 9. MReaEn EPggrShOeSlTGuhdiUcykWEnietihssPoMFOnSesr su(rrme)fmroemr anMaltlasrd r "Table 10.frMoeman MBaoldyarWdeiRgeh (pg)rofoHaSdttcuuhdicynWgtiatinhdo14n-Day OKPFOS..S..uc rvivors " APPENDICES Appendix 1 ComiocfAaBtYSeS ccrccrrnnncdS Appendix I. Diotand Supplement Formulations... Appendix I. DictPreparation rn $0 Appendix IV. The AnaoflPFOSyinAsviainDiset... ST Append V. DiaoFTgestrLayaoutm ccs Appendix VI. Reproductive Parameters and Replicate Knifiction... @ Appendix VIL ReApdrulotdaBtodiyonWeSitgahtyw(igthroFmOS.Mallard nT Appendix VII. FReeed CponrsuompSdttiuoudnyc(wgibttihsiPddRoaOyS)n.f.r.om a Mallrd ere] Appendix 1X. NIinadlilvairdduaRlepGrroodsuscPtaitohnolSotguidcyawliOtbhsPrvRtOiSons rom a nn $5 Appendix X. ARdueltpLirveroWdSetiuguhdctyswt(ipihfProoRmOnSa.M.a.llard eT Appendix. XI. HiSOpaROOgYREPO crv w [ERE] Wildlife International, Ltd. 5 Project Number454-109 `TABLE PAGE OF CONTENTS Appendix XIIA. Appendix XIIB. Appendix XII. Appendix XIV. Appendix. XV. Appendix XVI. Appendix XVIL Appendix XVIIL Appendix XIX. Appendin. XX. The Analysisof PFOSinAdultRed Blood Cells and Sa... 128 The Analysis ofPFOS in Adult and Juvenile Live and era. In RReperopducrtiovedSPeuryfocrwmitatnhicPeRobOynSP.en froameMeallard 158 RNeiparloadrudctRievperPoedurcftoiromnaSnucedbyywWietekPFaOnSd enfrom a 1 fgrgomSaheMlallTlhaircdkRneepsrsoMduecatsiuorneSmelnyt (wimtmh)PpFerOSenbyWeeki 193 MfreoamnaHMaatlclhairndgRBoedypWreigohStdt(u9udpyecwritPtehinPRboyOSnWeek ccc 195 MWeeacnk 1fi4o-mDayMOaIldlaSrudrvRievporrodBuocdtyioWneSitgudhy8w)iphrPFPOeSn,by 07 ORffespprinrg LoivdSertuuWdecyigwthittsih(PogROfnSr.om a Mallard 199 CH0SA yPS ROSOL.rrrerrnns 200 Personnel Involved inStudy. ns 202 GAL 00 Wildlife International, Ltd. Project Number454-109 "SUMMARY STUDY: PFOS: A Reproduction Study with the Mallard SPONSOR: 3M Corporation `WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER: 454-109 TEST DATES: SExtpuedryiImenntialtSita-arJtta-niuJoaarnnyua2r2y,2230,021001 FPihrostoEsgtgmsuSleattionAp--riApri13l,4,20200101 `AAdnaullyttTicearlmiTneartmiionnat--ioJnun-eA2p0r-i2l1,112,0200102 BEixopleorgiimceanltTalerTmeirnmaitniaot--nioJ-nulAyp2r6i,l 21010,12002 TESTANIMALS: Mallard (Anasplayrhynchos) AGETEST ANIMALS: 24 weeks ofage at the nitionofthe test. `SOURCE TEST ANIMALS: W1h1i3stWlaisnhginWgilnogns,StIrnce.t HUaSnAov.er, IL 61041-0509 NOMINAL TEST CONCENTRATIONS: 0, 10,50, and 150 ppm .. RESULTS: Tefhfeercetswueproenracdautltmebnotdrylawteeidghmtorattaltihteie5s0, aovnedrt15s0igpnspmof4.t1o.xitcesittycoanncdertarteiatomnesn.tseTlhaetreed wcoenrceentarlasotiotnr.eaTthmeent5e0laantded15e0ffpecptms .u.pontestfecodncceontnrsautmipotnisownerate ttehermi1n5a0tepdppmrio4r.oetshte roefprtoodxuiccittiyv,e oprhastereoafmthentteels.ateTdhereeffwcecrse no reament.rclated upon body weight, mortalities, overt Signs feed consumption or rinecprreoadsuecdtiviencpideernfcoermoafncsemaatlttheest1e0s pfoprmmaal.esteisnt cthoence1n0trpaptimona.. Htoewstevceonrc,entthrearetwioanstahant cnoanycehntarvaetiboene,ntehleatneod-oobsetrrevaetdm-eenftf.ectBacsoendceonnttrhateioenffcfotrs moablslearrvdesd eatxptohsee1d0ptopPmFa.OStesitn dtuheridnigetthiwsasstuddeyt.ermined to be less than 10 ppm a.., the lowes! concentration tested Gada Wildlife International, Ltd. Project Number454-109 "0 INTRODUCTION his study was conducted by Wilde Intemational, Ld. for 3 Corporationa the Wilde Incr national, Li. avian toxicology facility in Easton, Maryland 21601. The biological portion ofthe es was conducted from January 23, 2001 until uly 26, 2001. Rave data generated at Wide Intemational, Lid and a copyof he final report are filed under Project Number 454-109 in archives located on the Wide Iotermational Lid se. Biological speciremsoerendsa 3M Corporation, St.Paul,MN 55133 onscTIvEs The abjectiveofhs study was fo evaluat the effects upon the adult mallard (Anasplayrhynchios) of itary exposure o the potassium salt of Peflorooctane Sulfonic Ack (ereafler refered 1025 PFOS) over a period of approximately 21 wks. Effects on adult heal, body weight, and feed consumption were evaluated. In addition, the ffcts of adult cxpasur o PFOS on the numberof eggs id, erly, embryo. viability, haichabily, offpring survival, and egg shell thickness were evaluated Histopathologicalexamination ofselected tissues and analysesof lood nd ise samples were also used 10 evaluat the efects upon alts exposed 0 POS and hen offspring EXPERIMENTAL DESIGN Mallard (80 males and 80 females) wee randomly distibuted into one control group and three treatment groups. The test concentrations wer selected in consultation with the Sponsor, based upon the susof a plo reproduction study (Wilde International, Ld. Projet Number 454-105) and additions oxic information provided bythe Sponsor. PROSTreatment Groups Group Nomimal Concentration Pensper ExtaPons- BidsperPen a Growp __ematology_ Mates _ Females T (Contro)0 6 s 11 2 10 16 4 )1 3 0 6 4 )1 4 150 1s 5 :1 Gali Wildlife International, Ltd. Project Number454-109 ie Each treatment and conel group contained 16 pais of birds with ne male and one femal per pen. Four addons pins of birds were added t cach tt lve (or a total of 20 pais pe eve) nd mintained fo collection of blood samples during the es. Thre treatment groups were fed dicts containing either 10, 50,o 150 ppm ai. of PROS. The comol group was fed diet comparable to the ncatment groups, but without he aditon ofthe fest subsance . Because overt signsoftossty and came elated mortalities were noted at the 150 ppm ai. est concentration, the dietary concentrationfoth 150 ppm a. resent vel was reduced 0.20 pm of PROS at the begining of Week 3of the est. The 20 ppm a.. evel was terminated at the beginning of Week ofthe test du 1 he limit recovery ofbirds a his est concentration. Fo poring purpose, he 150120 ppm .. test concentration will be rere (08 he 150 pm ai. coneemraton. Aduls in he 50 ppm .. test concentrations were chanizd a th end of Week 7 of th fest du fo rcatmentclated mortality and over signsoftoxicity. Therefore, the test was completed with the control group and the 10 Prmai. test concentration nly: AN adult binds were observed daily throughout th test for signs of toxicity or abnormal behavior, Adult bod weights were measured at est inion, on Weeks 2,4, 6,8, and a adult termination and feed consumption was measured weekly houghou the fest. At the beginning of Week 11, the photoperiod was increased 0 induce eg production. Following the ir of egg production, eggs were st weekly for incubation, Weekly, eggs were selected by indiscriminate daw for eg shell thickness messurcment and all remaining eggs were candied prior (0 incubation to deect egg shell cracks or abnormal eggs. Eggs were also candied twice during incubation to detect infertile eggs or embryo mortality. On Day 24 of incubation, the cggs were placed in a hatchr and allowed to hatch. Once hatching was completed, hatchlings were removed from the hatches and the group body weight of the hatchlings by pen was deteined. At 14 days of age, th average body weight by parental pen ofall surviving offspring was detenmined. Upon completion of the test, atistcal analyses were performed fo determine statistically significant differences between groups. Chai Wildlife International, Ltd. Project Number454-109 ne MATERIALS AND METHODS The study was conducted according to the procedures outlined in the protocol, "PFOS: A Reproduction Study with the Mallard". The protocol was based on procedures outlined in the Environmental Protection Agency's Registration Guidelines Pesticide Assessment Guidelines, FIFRA Subdivision EHazard Evaluation: Wilde and Aquatic Organisms, Subsection 71-4; OECD Guideline 206; and the ASTM "StandardPracice for Condueting Reproductive SueswithAvian Species" (1.2.3). Test Substance The test substance, PFOS, was recived from 3M Corporation on October 29, 1998 and was assigned Wildlife Intermational, Ld. identification number 4675 upon receipt. The tet substance was a white powder and was identified as: FC-95; Lot 217. The test material had a reported purty of 86.9% and expiration date of August 31, 2006 (Appendix 1). The test substance was held under ambient conditions in locked storage a the Wildlife Intemational, Ld. facilis in aston, Maryland. Concentrations of the test substance in the diet were adjusted to 100% active ingredient. Therefore, dictary concentrations are expressed as parts per million active ingredient (ppm a.) in th die. Test Organisms Approximately 84 pais of pereared mallard were purchased from Whistling Wings, Inc. 113 Washingion Street, Hanover, IL 61041-0509, U.S.A. At the start of acclimation, the mallard were appasenly healthy and phenotypically indistnguishable from wid type. The birds were from the same hatch, approaching ther ist breeding scason and had no been used in any previous testing. At the start of acclimation, a random number gencrating function in a spreadsheet program was used to randomize pen assignment for cach bird. Immediately prior o test nation, all potential study birds were examined for physical injuries and gencral health. Birds that did not appear heathy, cither due to injury or inability o acclimate to laboratory conditions, or were outside the weight range for th test, were excluded from the study. All birds were 24 weeks of age at test initiation (fist dayof exposure to test diet) and ranged in weight from 781 10 144 grams at test initiation. Sex of the birds was determined by a visual examinationof the plumage. Identification Adult birds were identified by individual leg bands, ach pen was identified with a unique number, and groups of pens were identified by project number and concentration. All eggs laid during the study cna _ Wildlife International, Ltd. Project Number454-109 se were marked with he pen nomiber using permanent nk markin penfo detifcatin. Hachlings were identified by wing bands so tht they could be traced 10 ther parental pn oforigin. Avian Feed and Water AN adult bids and thei offspring wer given feed and water ad libitum during scclimation and (esting. The basa dit fed o both adults and offspring was formulated to Widife Intemational, Lid specifications by Agway Inc. (Appendix 1. The basal ration contained a least 27% protin and 2.5% ft, and no more than $% be The basal dit contained spproximately 1.1% calcium, derived from fecdstas and the 09% Jimeston usd in the formulation ofthe basa icby Ay. While this evelof calcu is sufficient for grove and maintenance rations, additional calcium is required in the ation of brcding birds for gg hell formation. Therefor, an additional $% (w/w) of limestone (approximately 38.5% Ca)wasadded to he basal diet for the aduls. This raised the calcium level in the diet for the breeding birds to approximately 3%, lightly sbove the minimum recommended for quail (2.3%) nd mallard (275%) (4) Ofipring recived basa die wihout test substance and vithout the addition of 5% supplemental estar. Water was supplied by the town of aston public water supply. Neither the aduls nor offspring ecsivd any formofmedication i th fed during he test. Fe and war were analysed periodically in accordance with Wilde Intemational, Lid. Standard Operating Procedures Dict Preparation Test dicts were prepared by mixing PFOS iio a premix tht was sed for weekly preparation of he fina dict, Control dit and cach ofthe thes raid dts wer prepared weekly beginning on January 23,2001 and presented t the birds on Tuesday of cach week. Ditay concentzatons were adjsted for purity of th test substance and ae rested as parts pes million active inredint (ppm 1). Deals of the weekly preparation of est and control dies ar shown in Appendix I Diet Sampling Homogenciyofth est substance n the diet was evaluated by collecting six samples from cach of he tested dicts and one sample from the convo diet on Day 0ofWeck 1. Samples were collected from Cadi EE -- Wildlife International, Ltd. Project Number 454-109 ae th tp, middle and brim of he left and right sections of th ising ves, Conrl nd rstment group dit samples were ls collet fromthe fed troughs an Day 7 of Week| 1 sess stably ofthe Let substance der actual tes conditions. Additonal, a sample was collected from the control and estmen group dis when available, during Weeks 2,3, 4,8, 12, 16 and 20 ofth et to masurcerity Lest concentations. The die samples were stored fiozen of transferred immedigely 1 the Wie Incrmational, Lid. analytical chemisty cityforanalysis Analytical Method The method usd fo th analysis of POS in avian is ws based pon methodology developed at Wid Iemasonal, Lid. and ented "Analytics Method Verification or the Determination of PFOS in Avian Die" (Wilde Inemationa, Li. Project No, 454C-110). Thi modifications from te ied methodology were incorporated nto the preset study. These changes were (1) the use of high performance liquid chromatography with triple quadrupol mas detection (HPLCMSMS) ater than ingle quadnpol mas deestion 1 reduce poenil mati. ieferencs, (2) the climination of he inna standardization procedure because an propriate intemal standard could no be iene, and (5) the use fashore (50 mm v5.100 mn) natal chromatography column 0 expedite alysis me. Samples were extracted with methanol, vacuum fered, did in ethan, and then ited n a P50F:O5S0 mmeetthhaondolo:logNyA.NOApumreet"howdatfelrodwilcuhtairotn isolpvreonvtidseodthiant AtphpeyenfdelilxwIiVt,hinFitghuerecal1.ibraCtoinocnernarnagieoonfstohef PROS in the standards and extracts of the samples wre deermined byreverse.phase HPLOMSIMS. sing a HwltPackard Model 1100 High Pestormance Liquid Chromstoraph incfacd witha erin TElhmeemraAssPIsp3e0ct0r0omMeatsesr wSapseecqturoimpepteedr owpietrhatePderiknimnu-lEtlimpelreTruebaoctlioonnSmpornaiytoiroinngso(urMcRe.M)HPdeLtCectsieopnaramtoidoen.s wre achieved wing a Keystone Betas Co analytical column (50 mn x 2 mm LD. 3m parce sie) itd with a Keystone ave Cx Guard Cridge (20 mmx 2 mm LD). Th inseument parameters ae summarized in Appendi IV. Table | of 4 C1a.l0i0brmatgionasLtandasrodcskwesreoporenparoefd PinR5O0S:50inmemtehtahnaonlo:l.NANTOhpeurcealbwaaiteornbystaapnpdraorpdrsi,atreadnigliuntgionisn concentration fom 0.00100 100.0100mg aL, were analyzed with each sample set. The sme and mst prominent pe response for PFOS was uiized fo monior PEOS i all calibration, quality cortol, and ent - Wildlife International, Ltd. Project Number 454-109 se study samples. No amp was made fo quaniify PFOS on the bass of individual omer components. Linea regression cquatons were generated using the peak area responses versus the respective concentrations of the calibration siandards. An example of a calibration curve i presented in Appendix IV, Figure 2. The concentration of PEOS in avian feed samples was dtemmined by substituting the peak areas ito the applicable linear regression equation, Typical fon chromatograms of low and high calibration standards ar shown in Appendix IV, Figure 3 and 4, respectively. Examples of equations usin calculations are presented in Appendix IV, Table 2 "The instrament limit of detcton (LOD) for these analyses was 10.0 pg on-column, calculated from {he product ofthe nection volume (10.0 hl.) an the lowest standard concenation 1.00 ug iL = 1.00 pL. The method limit ofquantiation (LOQ) for these analyse was 2.00 ppm .., calculateasd the produc ofthe lowest calibration standard analyzed 0.001 mg :L) snd th averal dition factorofthe mari blank sample (2000 L/Kg), Examples of calculations are presented in Appendix IV, Table 2. Along with the sample analyses, seven mtx blanks ers analyzed to determine possible imerfirences, No interferences were observed at or above the LOQ during the sample analyses (Appendix IV, Table 3). A typical fon chromatogram ofa mati blank i presened in Appendix IV, Figures vin dict samples were fortified at he following levels and analyzed concurrently with the samples to determin he mean procedural recovery. The fortification levels were 5.00, 500, and 200 ppm a. for sampling intervals Week 1, Day 0 Week 1, Day 7; Week 2, Day 0; and Week 3, Day 0. The fortification levels were 5.00, 250, and 100 ppm 8. fo sampin imerval Week 4, Day0. The fonction evls were 5.0 and 25.0 ppm ., fo sampling intervals Week 8, Day 0; Week 12, Day 0; Weck 16, Day 0; and Weck 20, Day 0 The method yielded mean procedural recoveries of 101%, 101%, 102%, 100% and 99%, respectively, for he 5.00, 25.0, 500, 100, nd200ppm .. fortification levels (Appendix 1V, Table 3). `Sample measured concentrations wer not corrected for the mean procedural recoveries. A typical on chromatogram of mai fortification is presented in Appendix IV, Figur6e. cra ---- ttret-- Wildlife International, Ltd. Project Number 454-109 "t6- Study Phases The primary phases of he study and thei approsimte durations were I Acclima3wieeoksn. 2 Prephotosimulaion- 10weeks, 5. Precgg laying (with photostimu-a0twoeenk)s . 4 Eaglaying- Approximately 10 weeks. 5. Postadut teminaton (final incubstio, hatching, and 14-day offspring rein period) - 5 weeks, HousinHgouasnidngEnavnidrohnumsebanntdarlyCpornadcititcieosnswer condictd 503 to adhe to the guidelines csablished by {he Notional Rescarch Council (5. The adult birds were housed indoors in bates of pens manuficurd by Safeguard Products nc. (Model No. S355), messing spproximacly 75 X 90 X 45 om igh he pens wee construct of inyhoated wis mesh. A disgram of the fst yout is presened in AppendiVx. Each pen was capped witha fed trough. Weekly, sufficient fed fo he fecding period was placed in he ough for cach pen and presented th bids. During he fein period additional feed was weighed and added1 th rough as psd. Water was suppliedby ippl-ype teers. Oly birds asocited with tis td were mained in the study room in oder 0 avoid excessive disturbances. The average temperature in head mallard study room durin the course ofthe Lest was 217 IC (SD) with a average relative humidity of 40% + 19% (SD). The i handling System n he study room was designed 10 ven up 0 15 room air volumes cry hou and replace hem withfresh si. The photoperiod in the adult mallard room ws maintained by 3 time clock. The photoperiod during acclimation and the fs 10 weeks of the tst was cight houso lss of ight per day. The photoperiod was increased 0 17 hours fight per day at the begining of Week 1110 induce egg laying and was maintained a that lengih unl he dul bids wre cuhanized. Throughout he es, th birds received a meanofproximately 188 ux (- 17 candies) of llmintion provided by furescen lights ns closely approximated non-day sunlight cn --- Wildlife International, Ltd. Project Number 454-109 i Observations "he est bis were acclimated the facilities an stdy pensfo approximately 3 wecks prio 0 initiation of the tes. During acclimation all birds wer observed daly. Birds cxibiting abnormal behaor vdeiilaoinrg physical juries wer no usd forthe est. During th study, ll adult binds were observe daily for signs of toxicity or abnormal behavior. Addionaly, al ffpring were observed daly {rom hatching until 14 days of age. A record was mainaited ofall mortalities and clinica observations. Adult Body Weight and Fecd Consumption Adult body weights were measured atest nition, on Weeks 2,4, 6,5, and at adult termini. Boy weighs wer not measured during egg lying because ofthe possible adverse efcts handing may haveon egg production. Feed consumption for each pen was measured wckly thoughout th st. Feed consumption was determined by weighing the freshly led feeder on Dey 0, recording the amount of any additonal die add during the week, and weighing th fcder and sensing fed at the ndofth fcding period (Day 7). The amount of feed wasted by the birds was not quantified, sinc the wasted feed was normally catered and mixed with water and excreta, Therefor, sed consumption is presnted a an estimate of {oe feed consumption. Adult Blood Caletion Four addons pais of adult birds were added 0 sch st evel 1 be msinainefo collection of bod samples during he test These pairs reise th same dit and were handled in he same mance as other pais in thei test group. However, because of the potential ses of the blood collection procedures, dat rom the four additonal pais were nt sed 10 evaluate tssmentrlated eft. Blood amples wer collected from birds in these designated pairs, when possible, at five week intervals broughout the test. A the ime of dul termination, blood samples wee cullcid from all surviving binds, when possible, prior fo culanasa, All blood samples were separated ino serum and hemacyespltlet, sored frozen ad shipped 1 he sponsoro sponsors designatefo possible analysis. Adult Necropsy and Tisue Collection When the remaining birds from 150720 pm a. raiment group we euthanized at the end ofthe Wek 4, no ncropsics were performed. All afer dul birds tat did or were cuthanized during the naan Wildlife International, Ltd. Project Number 454-109 "ise course of the study were subjected 10 gross necropsy. At the conclusion of the exposure period, all surviving adult birds were cuthanized by cervical dislocation, necropsed, and disposed of by incineration At th time of necropsy, tissues were calleted for histopathological examination and possible analyses. When availble, samples of gal bladder, liver, proventriculus, kidneys, brain, gonads, bursof Fabricius and adipose tissue were fixed in 10% buffered formalin and shipped to EPL in Hemdon, VA for histopathology. When avilable, sampls of bile, lve, proventriculus, Kidneys, brn, reproductive organs and adipose tissue were stored frozen and shipped to the Sponsor for possible analysis. Fecal samples were also collected, when available, from the lower digestive rac and stored frozen for possible analysis. Keg Collection and Storage Egas were collcted daily from al pens, when availble, Eggs to be incubated wee washed to reduce the possibilty of pathogen contamination before storing them in the cold room. Eggs were washed in a commercial gg washer (Kul Egg Washer) witha chlorine based detergent (Kuhl Super CD). Water inthe washer was warmed fo proximately 45C. The eggs were placed in the wash water and sosked for approximately 15 seconds. The washer' circulation motor was then tured on for approximately tree minutes. Th eggs were removed from the washes, allowed t cool to approximately room temperature and rinsed with fresh wate. The gs were then stored in cold room unl incubation. "The cold room was maintained at a mean temperature of 13.5C 0.2C (SD) with a mean relative humidity of approximately 69% + 6% (SD). Groups of eggs were identified by a alphabetic lot code Alleges id in weeklyinerval were considered as one lo. Candling and Incubation A the end of th weekly interval, all eggs were removed from the cold room, counied and cggs elected by indiscriminate raw for cg shell thickness measurement. The remaining eggs were candled With a Speed King (Model No. 32) cgg-<andiing lamp to detct cg shell cracks or abnormal eggs Crackedo abnormal eggs were recorded and discarded, A eggs no discardedo used for cg shel thickness measurements wee placed in a Petesime Incubator (Model No. $P20). In the incubator the temperature as maintained at an average 37.5C = 00C (SD) with an average wet bulb temperature of 30C + 03C (SD) Gelatve humidity of approximately 60%). The incubator was equipped with a pulsator fan and blades that produced a mild 610 Wildlife International, Ltd. Project Number454-109 "9 bresthin air movement designed to cline inecabinet temperature and humidity variation during incubation. In oder to prevent adhesion of the cmbryo to the shell membrane, the incubator was also quipped with an sutomatic gg rotation device, designed to rotate the eggs from S0offof vertical inane direction t0 50offof vrica nth apposite direction (otal ae of rotation was 100) evry two hors through Day 24ofincubstion. Es wee candled on Day 13 or 14 of incubatioton determine embryo viability and anDay 21 to determine embryo survival Hatching and Brooding On Day 24of incubation, the eggs were placed in a Peerime Hatcher (Model No. SP-GH) and allowed to hatch, Pedigree baskets construct of gabanized tcl wire mesh were used fo keep atchings separated by parca pen of arin. Eggs were not rotted in the hatcher, The average \emperatre i th hatching compartment was 372C + 00C (SD, and the average wet bulb temperature was sed 0 3.8% 02C (SD) relative humidityofapproximately 80%). A atclings, unhatched eggs, and eg shells were removed from the htcher on Day 27 or 28 of incubation. Thegrboodyuweipght of the surviving hatchlingsbypeswas determined. Hachlingswere wing banded for entfcation by pen of riin and then oinly housed according to the appropriate parental concentrstion grouping in brooding pens nil 14 days ofage. The hatchlings were fed untreated ie without headin of 5% supplemental limestone. A 14 days of age, the average body weight by parental pen of al surviving ducklings was determined, The ducklings were suhanized with carbon dioxide anddisposed ofby incineration, Hatching were housed in bars of brooding pens manufactrsd by Safeguard Products, Ic Each pen measured approximately 62 X 92 X 255 cn high, The wll, floors and ceilings of each pen were constructed ofvinyosed wir mesh. Thermostats inthe brooding compartment of cach pen were eto maintain a temperature of proximately 35C from the time ofhatching util he birds wre ive to seven days of age, when the temperature was adjused msintin temperature of approsimatly 29C. The average ambien room temperature was 242 + 12C (SD) with an average relative humidity of 7754: 15% (SD). The photoperiod or the hatchlings was mained by ime clock at 16 hours of ight por day. nant Wildlife International, Ltd. Projet Number 454-109 "20 Offspring Blood and Tissue Caletion Prior to euthanasia of te ast group of offpring (Lot J), blood samples were callctd from 10 offspring in both th con group and 10 ppm .. resiment group. All blood samples were separted into serum and. hemaeyespltlet, stored frozen and shipped 10 the sponsor or possible analyses, Addidonally, tissues from the 10 offspring in each group were collcied for histopathological examination. When availble, samples ofgal ladder, lve, proventriculus, Kidneys, brain, gonads, bursa of abricus and adipose tissue were fixe in 10% buffered formalin and shipped to EPL in Herndon, VA for histopathology. When availble, samples of bil, liver, proventriculus, kidneys, brsin, reproductive organs and adipose tissue were stored frozen and shipped 1 the sponsorfo possible analyses, Fecal samples wer alo collected fom pans below the loo of broodes housing Lot J offspring and stored frozenfo posible analysis. A single compositc sample was collected (or the control group anda single composi sample was collected or the 10 ppm .i. reste group. Egg Shell Thickness Measurements and Egg Components Callection Weekly throughout the egg laying period ane gg was cllcied, when availabe, from eachofthe test pens. Eggs were collected from odd numbered pens during odd numbered weeks (13.5, etc) and from the even numbered pens during the even numbered wecks (26, tc). The eggs fiom weeks 1, 2 and 10 were separated ino albumen, yolk, shel and membrane and all components were sored frozen. Ege collet during weeks 39 were opened at th waist and her contents removed. The cg contents wre separated to sibumen and yolk and the yolk was store frozen. Al gs smples wre transfered 10 the Sponsor for possible analysis. The shells from Weeks 3 through 9 were alowed to sir dry for at Least one week a room temperature prio 0 being measure for thickness. The average thickness of the ried shell pus the membrane was determined by measuring iv points around the waistofthe gg using amicrometer. Measurements were made to the nearest 0.002 mm. Statistical Analyses Upon completion of the test, an analysis of variance (ANOVA) was performed to detemine srsically significant differences between groups. Dunnett's malipl comparison procedure (67) was uscd to compare the thre treament means with the control group mean and assess the statistical Significance ofthe observed differences. Sample units were the individual pens within cach experimental group, except adult body weights where the sample unit was the individual bird. Percentage data were cxamined using Duets method following arcine square foot transformation (sce Appendix V1 for Rann e--r ------------------------------------ Wildlife International, Ltd. Project Number 454-109 a reproductive parameters). Wilh he exception of Tver weigh, statistical analysis of the data was performed wing Avian Reproduction Data System (ARDS) Software a validated software package developed by Wife Intemational, Lid. (9). Liver weights were compared by Students tet sing Microsoft Excel. Eachofthe following parameters was analyzed statistically: 1. Adult By Weight - Individual body welght was measured attest nition, Wecks 2,4, 6, 8 and a adult termination. Sisal comparisons were made between th control group and each wreamen group at each wighing interval by scx. 2. ACdexuoalmtniFnseeedudbmy ppetn wieeoklny d-uFrienegd tchoentseustm.ptSiaotniseixcparlescsoemdpaarsisgornasmsweorfefmeeaddepebretbiwredenpetrhedacyonwtaosl and cach wsament group. 5. Fags Laid of Maximum Laid -The number of eggs laid per female divided by the largest number of cg id by any one female. This transformation was ued 0 comer the umberofeggs sid 08 percentile ala es tha or cqal 1 10, 4 FagsCraofEcggskLaed dThe number of eggs determined by candin to be cracked divided by he amberofeggs ad, per pen. 5. Viable Embryos of gas Se-Tthe mumber ofviable embryos a the Day 14canding was divided vy the number ofgs ct,perpen 6 Live 3Weck Embryos ofVisble Embryos - The number of live embryos a the Day 21 candling was divided by the umberofvile bryos, pr pn. 7. Hatcofl3-WiecknEmgbrysos - The numberofhatchlings removed from the hatch was divided bythe umber of lve week embryos, pe pen 5 14Day Old Survivors of Hatchling - The mamber of 14-day ad survivors was divided by the umberof hatchlings po week, by pn. 5. Hatchlings of Eggs Set - The number of hatchlings was divided by the number of eggs set per week, by pon. 10. 14-Day Od Survivors of Eags Sei The number of 14-day old survivars was divided by the mmberof gg set per week, by pen 11. Hatchling of Maximum Set - The number of hatchlings pr male divided by th largest number of eggs st from any one female. Thi transformation was used to convert the number of hatch ingstoapercentile value cual 0 ores than 100. nang Wildlife International, Ltd. Project Number 454-109 - 12. 1S 4-Du ayOlrd viv of Mo axir mumsSet - The number of 14-day old survivors per pen divided by he largest umberofeggs st. 13. Ex Shell Thickness The average gg shel thickness of indiscriminately selected eggs per pen was measured 14. Offspring's BodyWeight - The group body weights of surviving hatchlings and 14-day old `survivors were measbuyprareentdalpengroup 15. Liver Weight Individual Tver weights were measured at aul termination and ot termination of selected LoJ offspring. Statistical comparisons wers madeby sx betwee he control group and 10 ppm a. group. RESULTS AND DISCUSSION Mature mallrds recived PFOS at nominal dietary concnteatons of 10,0, and 150 ppma. Du 0 extensive overt signs of toiclty and satmentslated montalies in the 50 and 150 ppma. testment groups, the experimental design was modified. Adults in the SO ppm a.. test concentrations were cuthanized at the end of Week 7ofthe test. The dietary concentration for the 150 ppma.i. treatment level w{earsmirneadiuecdeda toth2e0bpepgimnna.ii.ngofofPWFeOeSkat$thoefbtehgeintneisngduoef W(0eetkhe3liomfittheed tresetc.ovTerhye o2f0 bpirpdms ta.i. tliesvellewveal,s Therefore, the test was completed with the control group and 10 ppm ai. test concentations oly. The convo group was fed diet comparable to the rctment groups, but without the addition of the test substance, Diet Analytical Results None of the control samples showed ary indication of he presence of he fest substance or of the presence of o-luing subsiances a th characteristic rtion ime of the test substance. Dit samples were cllcted fom the 10,50 and 150 ppm a. test concentration on Week 1, Day 0, and were analyzed 10 evaluat the homogeneity ofthe es substanc inthe diet nd to verlfy test substance concentatons. Means and sandard deviations for he tre fest concentrations were 9.75& 0313 ppm ai, 50. 0981 pom ai. and 147 + 431 ppm .., respectively. The cosficients of variation were 3.21%, 1.96% and 293%, respectively. These values eproscnicd 98%, 100%, and 98% of nominal concentations (Appendix IV, Table 4) GnLant Wildlife International, Ltd. Project Number454-109 a. Samples collected throughout the test o verify tet substance concentrations for the 10, 20, 50, and 150ppm ai. diets had means and standard deviations of 10+.0.5253 ppa.m 20.8 2.14 ppm a, 09 2.04 ppm a, and 161 15.6 ppm ., respectively. The cosfiiens of variation were 544%, 10.3%, 401% and 9.66%, respectively. These values represented 102, 104, 102, and 108% of nominal concentrations (Appendix IV, Table 5). Analysis ofdiet samples collected from fegders aftr being held at ambient temperature fo7r days aversged 100%, 101% and 97% ofthe Day0 values for the 10, 50 and 150 ppm ai. tet concentrations, respectively (Appendix 1V, Table 6). A typical ion chromatogram ofa test samplei shown in Appendix IV, Figure 7. Mean weekly measured concentration for cach test group are presented in Table 1 Mortaities While there was one incidental moray in the control group, thee were no morals in the 10 Pom a. restment group. Three incidental moraiies and tentreatmentrelated morales occured in he 50 ppm a. treatment group snd four treatmentrelated moralies occurred in the 150 ppm a treatmentgroup "The singe incidental mortaliityth conirol group was amale (Pen 210) ha did during Week 15 ofthe est. Gross necropsy ofth drake revealed foot lesions on both eet. Internally, emorhages were noted in the myocardium, th abdominal aie sacs were cloudy andth spleen was enlarged and motiled. Additionally, the ivr was fim, motiled and ple, th kidneys wer slighty pal and a small amount of lear uid was noted in th abdominal cavity. Necropsyofthe bird' penmate at terminationof the adult portionofthe study revealead regressed ovary, but was otherwise unremarkable, Thee incidental and en treatmentrelated mortalities occurred in the 50 ppm ai. restment group and four ratmentrelated moraltes occurred in the 150 ppm ai. treatment group. Necropsy findings or ihe treatment related mortalities in the S0 and 150 ppm i. test concenraions were similar. Common findings included thin or emaciated condition with a loss of muscle mass and prominent kee, small specn and pale Kidneys. All sensing birds at the 150 ppm a. test concentration were euthanized at the beginningofWeel. Due fo th shorencd natureofthe expasure period, necropsis were not performed. Al remaining birds at the S0 ppm .. test concentration were euthanized atth beginning of Week of the tet, and necropscs were performed folowing cuthanasia, Necropsy findings are summarized in Table 2 Gai; Wildlife International, Ltd. Project Number 454-109 "ne Clinical Observations No over signs of toscty wer absrved at the 10 ppm .. est cancenration. With the exception of foo lesions and occasional lameness, sl birds inth control group and 10 ppm a. reament group were normal in appearance and behavior throughout th tes. 10 the 150 ppm a. and 50 ppm a. treatment groups, signsof oxilty were fis noted at the end of Week 2, One male inthe 0 ppm a. treatment group and several birds i the 150 pm .i. treatment group hibited signs of toxicity induced by the stress of handling during the Week 2 body weight measurements. In several cass, birds convulsed during body weight measurement. Signsof toxic in both the 50 nd 150 ppm a. resiment groups continued to be observed un the birds were euthanized. Clic signs of toxicity observed included depression, reduced esctio (0 extemal mui (sound and movement), wing droop, los of ordination, ruffled appearance, chin ppearanc, lover limb woes, lethargy, crimtion and convulsions Adult BodyWeight There were no apparenttreatmentrelated effets upon adul body weigha he 10 ppm a. test concentration, and any difeences between the control group and the 10 ppm .. test concentration were not statistically significanta any of th body weight tervals. However, here were marked treatment. elated reductions in mean bods Weight a both he 50 and 150 ppm a. est concentrations, With the exception of the mean ml body weigh tthe Weck 2 interal, al differences between the contol group and bids in the 50 ppm a. resment group wer statistically significant atp <0.01. At he 150 ppm a. test concentration, mean male and female body weights wee satsicaly diffrent rom ihe contol group a p< 0.01a both the Week 2 and Week4 body weight nterals. Between the Week 2 and Week 4 body weight intervals, a weight gain was noted fo birds at the 150 ppm a. est concentration. However, the weigh gain observed was efletion of the reduction ofthe concentration of PFOS in the die to 20 ppm a.. during Week 3 of the test. Mean body weight measurements are presented in Table 3, snd Figures 1 and 2. Individual body weight measurements are presented in Appendix VL entang s _--_e ---, --s ,,--,--,--,---- Wildlife International, Ltd. ProjctNumber 454-109 "5. Adult Fesd Consumpiion There were no apparent trate elated effects upon fed consumption at the 10. 50 ppm ai. test concentrations. No saistically significant differences between th control group and 10 or $0 ppm a. tenment groups were observed at any ofthe sed consumption ntenals. However, rcatmentelated reductions in feed consumption were observed atthe 150 ppm a. est concentration during Wesks 1 and 2. The reductions were statistically significant at p< 0.05 during Week 1 snd at p< 01 during Week 2. The dicary concentrationfo the 150 ppm i. resent group was reduced fo 20 ppm .. during Week 3 oPfotmhei.testte.stTchoenrceenwtarastiaonstdatuirsitnicgalWleyeskigsni3fincadnt 4(pof<t0h.e0e1t).incMreeaasne ifneefdeecdoncsounmsputmipotniomnesatsutrheme1n5t0s(s2r0)c shown in Table 4 and Figure 3. Feed consumption measurements by pe ae presented in Appendix VIL Adult Necropsy and Liver Weights AN surviving adults in the contol group and he 10 ppm .. raiment group were subjected to 10s necropsy following adult termination. When compared othe control group here was an inressed `iWnhciicdhenceroafimmaelnetsrewliatthedsmfalflsesotusld(anpoptrobxeipmraetceluldeendg.th<Tw3o.5mcaml)esiinnthtehe1c0onptrpolmgra. orupesweernetnogtreodupwiftohr smal teste, compared to seven males in the 10 ppm ai. growp. All aber ndings were considered unrelated to ncatment. Necropsy findings re reported in Table and Appendix 1X. When compared weights of adul livers to the concol tthe 10 ppm group .. tet there were no concentation eamentetd ffcts Any difference between upon the mean the 10 ppm a. raiment group and the control group was not sstisialy significant. Mean adult iver weights are presented in Table 6, while individual adult iver weights are resented in Appendix X Histopathology No lesions considered ltd to test atic (PFOS) adiministaion were noted inthe iver, kidney, adipose sue, brs, and proventriculus of adult an offspring males and females, bursa of Fabricius in offping males and female, ovaries of adult and offspring males and fests of mule offspring. The increase in th numberofadult males nthe 10ppm a. ested group with reduced testicle siz (with or without coresponding microscopic observation), suggests (at PROS ay hve sccleatd erly post eproducive phase regression, normal physiological phenomenon, Any potential estar ffcts on ihe bursa of Fabricius in adult mals and females could not be determined since samples were not Chara -_-- Wildlife International, Ltd. Project Number 454-109 ao sofusbpmriitnegdcfoorudevanloutatbieond.efiInnitaivdelyod,etacniynpeodtendtiualottehsetxteensfievcetasuotolystihse athltbaprdedcelrudfedoamnaydualctusratned agnosis. 10 ppm a. The ew other ested groups changes in varios tisues were considered incidental of alt ano offspring malls rom control and and unrelated ost article (PFOS) dminstration. "The fll histopathology ror iprovinAippdendeixdXI . AdultLiver andSera Adult load samples (bth rd blood cels and sera) collected through Week 15 of the est were analyzed by Exygen Research. The results for anlyss of samples called for this study are reported ager with els for the nthe bobwihite study un concurently with the same (st concentrations. SthurmomugahryXXreIsVultsloocfatbedloidn AapnpaleynsdeisxpeXIrIfAo.rmeTdheforfuhllisresptourtdyfaorreanparleysseinsteodfibnloEoxdygseanmprleepsortbyTaEbxleysgeXn Resarch is located der Biygen Study Number 023.066, "Exracion of Potassium Peruorooctancsulfonse from Red Blood Cels and Serumfo Analysis Using HPLCElectrospMraasys Spectrometry". a. testBlcoonocdenatnrdatiivonrwsaemrplseispcpeodll0e3aM tEhnevtiimoenomfeanutllLtaebromriantaotriyonfooratlhysciosn.troSlugmrmouaprayndre1su0ltps pof s2er4andan1d0ltivoeruagnhaly1s5esloccoantdeudctinedApbpye3ndMixEnXvIiTrBo.nmeTnhtealulLlabroerpaotrotryfoarr anaplryessiesntoefdseirna3ManrdeploirvtrTasbalmepsleIs, collet at the time of adult termination s locate in 3M Environmental Laboratory report E0I-1256, Analytica Phase Report fos PROS: A Reproduction Study with the Mille Reproductive Results There were no reatmentrlted uffts upon reproductive performance at the 10 ppm ai. ts concenraion 3s compar to the conrols. When compared 1 the control group, there ere no sutsicaly significant differences in any of th reproductive parameters measured in the 10 ppm ai. est concentration. Summarcs ofthe reproductive data ar presented n Tables 7 and 8, and in Figures 4 and . Reproductive parameterbypn re presented in Append XI and XIV. enLany _--_-- Wildlife International, Ltd. Project Number 454-109 a. Eg Shell Thickness "There were no apparent treatment related effets upon gg shell thickness at the 10 ppm a. test concentration. When compared to the control group, there were no statistically significant differences in egg shell thickness in the 10 ppm a.. treatment group. Egg shell thickness data are presented in Tabl9e and Appendix XV. Offspring Body Weights "There were no apparent treatment slated effects upon offspring body weight atthe 10 ppm ai. test concentration. When compared to the control group, there were no statistically significant differences in the body weight of hatchlings or 14-day old survivors from the 10 ppm ai. treatment group. Offspring ody weight dataar presented in Table 10, and Appendices XVI and XVII, Offspring Liver Weights "There were no treatment elated effets upon the mean weights of offspring livers a the 10 ppm a. test concentration. Any difference between the 10 ppm a. treatment group and the contol group was not statistically significant. Mean offspring liver weights are presented in Table 6, while individual offspring liver weights are presented in Appendix XVIII. Offspring Observarlons There wre incidental observations of neck curl, weakened condition and footleg lesions and associated lameness, for offspring in both the contol group and 10 ppm a. treatment group. At the 10 PP i. test concentration, a general observation was made that th offspring from Lois C and D were sluggish, witha slight reduced reaction o extemal stimuli. None of the offpring in he Lots proceeding or following LotsC and D were noted a sluggish or with reduced reaction to external simi, Due to the isolated natureof th clinical signs oftoxicity noted, these observations were not considered treatment elated. All other offspring were normal in appearance and behavior throughout the est Offspring Liver and Sera Blood and liver samples collected at the timeof offspring tenmiationfo the control group and 10 PP i. test concentration were shipped to 3M Environmental Laboratory for analysis. Summary ress of scra andliveranalyses conducted by 3M Environmental Laboratory are presented in 3M report Tables 1,2 and 10 through 15 located in Appendix XITB. The full report for analyss of sera and liver samples Gnang _--_--mm Wildlife International, Ltd. Projct Number 454-109 ne l"Alncaltyetidcaalt PhheasteimReepofojrutvefonrilPeOSte:rmiAnaRteipornoiducltociaotnedStnud3yMwiEtnhvtihreoMnmaelnlta"l.Laboratory report E01-1256, There were no feament elated concLusion mortals, overt signsoftoxicity,o weamentrlated effects uHopwoenvbeord,ythweerieghwta,seaendicncornesausemdpiinocnidoerncreeoprfodsumcatlivetepsteersfforomramnaclesiintthhe1010ppmpmaa..tteessttccoonncceennttrraattiioonn athnadt veaaytmBeanvteeblaetesdlaetffeedctfs urpaomneatd.ult Tbhoedrye wweeirgehrtaaitmethnet5r0elaatnedd m15o0raplpems,i.ovteersticgonnceonfirxtiionis. T0h0e1re50weprema.h rteastmceonnctesnlraatteidonesffweectrseftdrcionsmumpptriioon1atthhere1p5r0odpupcmtiav.e pehsatsecoofnhcenetratetsit.on.BaThse 5o0n ethxepofsetd 10obFcOrvSedinathtehed1i0epwpamadecerticeondcentrbaetieons, tthheanno1-0opbsperasie,dtehfeeltocwoenstcecnoatnocnfeontmalsltaredds during is sud, nro Wildlife International, Ltd. ES REFERENCES ProjectNumber454-109. | USuSb.divEinsviionronEm.enHtaazlarPdrotEevcatliuoantioAng:ency.Wildl1i9f8e2.andPesAtqiucaitdiecAsOsregsasnmiesnmts,Guisdueblsiencetsi,onFI7F1-R4A, Environmental Protection Agency, Officof Pesticide Programs. Washington, D.C. 2 O`rOgEaCnDizGautiidoenlifnoerfEorcToneomiscotCfoCohipeemirncaatlgiso.n aGnuiddeDleivneel2o0p6m.enPta.ris.1984. Avian Reprodiction Test. 3 ARmeeprroidcuacntivSeocSiteutdyiesfowritTheAstviinagn SapnedcieMs.ateArSiaTlMs St1a9n8d6a.rd ES1t0a6n2d-a8r6d. PrAancniucael fBoorokCoonfduAcStiTnMg Standards. Vol. 11.04.Philadelphia, PA. 15pp. 4 Me&rCoc.Iknc. 1991. The Merck Veterinary Manual, Mer&cCko. Rahway, NJ. 1832 pp. 5 NWaatsihoinnagltoRne,sDcCa.rchNatCioounnacliAl.cade19m9y6.PresG.uid1e24fopr. the Care and UseofLaboratory Animals. 6 wDiutnnheatCto,ntCroWl. Jo1u9r5.5.AmeAr.MSutlattiip.leAssCoocm.pa$r0:is1o0n96s-1P1r2o1c.edure for Comparing Several Treatments 7 Dunaon.ett, C:W. 1964. New Tabsfor MultipleComparwiithsCoontnrosl. Biomer2r0:i4c82s8 Wildif International, Ltd. 1994. AvianReproductionDats System (ARDS)Version 2 carte 0 PRives Fle 1, i: | i 3 | 2s : | C1t 2:| 5 || 1i| C Hi a. |, | || 55 g | 15]: gy is . LL i!n i =2 iis \ i | Ce v : i 3fh2f3s 2F iEETo 2 23| 13 - zi: FEN lg3 E3 EH i v i. Co os <25% i)z : /x np || | Hit ii j HiHi is i H iHf hiH i iP1 ali sl or ] i=]. ) Chain ER Project Number 454-109 Table 2 SummaryofGross Pathological Observations from a Mallard Reproduction Study with PFOS Adult Birds Found Dead/Euthanized during the Test Males Comal 50) Oppmai) ppmai p1p5m0a7i Females o50ma pp1m50a7i Numberof birds 1 n 2 18 2 Abrasion on mandible 0 1 o 0 0 Footleg lesions 1 3 o `0 0 Thin 0 4 0 3 o Emaciated 0 0 1 2 1 Lossofmuscle mass 0 1 1 3 1 Prominent keel 0 0 1 2 1 Breaanstdmluoosscelbe-lohoedmatoma 0 1 0 0 0 Hear - faced 0 0 0 1 0 Hear -pale 0 3 0 1 0 Heaercc-hpyetmcacthiiahle0morshages 1 0 0 0 0 Lef`tpllauqnuge-soynellmoawrgins 0 1 0 0 0 Lungs -pale 0 1 o 0 0 Abdominalai sacs - cloudy 1 o 0 0 0 Spleen - small andlor pale 0 7 0 5 0 Spleen -nlarged and med 1 0 0 0 Liver- firm, pale and moted 1 0 0 0 0 Abaduotmoilnyasliscavity - o ' 0 o z Abdaommoiunnatlofcacvliteya- fslmuaildl 1 0 0 0 0 Abdionmlienfalthocarvaictiyca-ibslaoocd 0 1 0 0 0 Gizbzialred-scaoinnteednt-s 0 3 0 2 1 Sma`lwlhiinttuelscteirnsep-oslylpls) 0 | 0 0 0 Intestinal contents -pasty 0 o 0 0 1 Inesinal contri - firm 0 1 0 1 o Kidneys -pale 1 4 0 0 ' -- Not remarkable: 0 ' ee 1 10 0 Robseervamtionabsirfdio bninUdseif5o0mpnpwhmig5c.h1.beltoscoanmcpelnetsrawteiornevecroleleecuteadnaiareodtartehpeoretneddof Week 7 f te(st.Grossnecropsy Rwemearinnoitnpgeirrfusmeidhatet1h5e0teprmpinmat.iotneosfttchonece1n5t0rpatpiomn.w.erescchoncaentzratUione. beginningofWeck 5. Gros eciopsics ee Gada] oe ------------------------ ProjectNumber 454-109 i xg 5g 1 Bole fares ko CE Bee eee 1 lh SavEI i RE T= 3% 82 i i 3 [EAL LANEI y ELt BE ovoes se scge | GEiEfE Sovran gE ae 11:1 IREIEIEIRIRIAR I LE i: tiff f Felld = 3 3 li15a%rs AR ae Project Number 454-109 i |: = oo M mT ---- B: 08c I$ .od . i 3 3z. | r TTTRE i ti8e og | CR [3 oH 2% feel i sh Bh pe, i 23| A -- "mE, FL:i 32%|| m pm -- :10 51 my { ik g 2] 2 m LF o mm--m ------ vBi%t8ggregesne J wworan Ace 3 i ;Poe0 . Cnet e-- mer ---------------------- ie Project Number 454-109 3 am JIT] #Bi E I E, 0 g om ki . 3i { 1x . Wo of:: EY a | TE tie 23Hg8c bfh oirBEeEL 2 3% RE i 5 &233 =BS HL i 2i?f FEE; [2* 5; 0- 53| qm= 25 ` i: & Tp Bf4 fggregenae hora soos Twas ;Poe0 ; Grin eee a5 ------------ Prot Number 456-109 3 #83 ' 2] 3 8 + Ads 8 - a 2 oo . 3 slog i sos o8 3stlo)is] os 8os 0 & fu{7p =F s =| 2 8 8 id Al 5 os Pll. 3 din 58 Proprio lass fi loseis fil loss oz zd & "ls oz oz & iH IE sii3 oieP: E3 LE EEors -- "36+ Project Number454-109 bY > M < . of x 5 :3 x : ~.. 2 ge ols un 4| EIe J{, oo*ii | s . 2 Yo. .3 is le Wi 8% >> 3 ou. C& - & Ten J - 3 CQ ge 3 swocuiesuv | nets -37- Project Number 454-109 Table S `SummaryofGross Pathological Observations from a Mallard Reproduction Study with PFOS `Adult Birds Euthanized at Test Termination Males Cponptmraoli) ppm0ai eee LE Numberof birds 15 16 Footeg lesions, 4 + Breast blister 0 0 Cranium - swollen and bruised 0 0 Heafrutid--pfeirlilceadrdial sac: 0 0 Spleen -enlargedandmotled. 0 0 Spleen -enlarged and firm 0 0 Liver - irm, pale and mottled 0 0 Abdominal cavity - Abdyoomliknraelmncaavnittsy - - - slight egg yolk peritonitis - - Abdominal cavity - eggyolk peritonitis - - `Abdominal cavity - slight old egg yolk peritonitis - - `Small intestines- area of hyperemia 0 0 Kidneys -pale o 0 Testes-small (~ 1.53.5 cm) 2 6 Rep`rmoudlutciptlieveflturiadctfi-lled cysts - - Ovary -regressing - - Ovary -regressed. - - Ova-cyrstyic follicles: - - Ovary - hemorrhagic follicles - - Not remarkable, 9 s Females Cponptmraoli) ppm0ai Ee 16 16 8 3 0 2 0 1 0 2 1 0 0 1 1 0 1 0 1 0 2 3 5 0 2 0 1 0 - - 1 3 1 0 4 4 1 1 1 3 2 0 "Grossnecropsy observations forbirdsmaintained forcollectionof bloodsamples during the test are not reported. cnt n Nees 5009 2is) 2 -2 . ` g 5gl0a g 28 2183 i z 23 . 33 2B23gl 2 i 1g: = i il 28 ds 5 + 1 iosl 3BEF: _C.:EF Co 1 ii i : 2: i o nH1 ih iif 3 TECHi | 28% nts ---,,-------------------- i. rsNumer 454109 i 3 a 8 LL 03 Ife si Ei Pf i reREraRRcre i HERRERAEEEEEE 3 Pll gi 3 1i?3 8 sl 2% upl Hx E3 bs alLi]2 si3 Fip sl iBihgrelBDiiiSepsit) Rs Bl o3igaspd5ciain ss#dl4z nian _--,,.. -- --. wo Projct Number454-109 * nfis 3 t i 8 i "ly 1I ge #Bi 53 55 28 I] Bil; o8fs| EE IN Oa oi ceSv SENN, Erina n nny) pis 2 8 2 8 | nda a Pc Nome 450.109 I P$ E |slsessesesesss| Pilg is ; - 1}zs i 1 i: |i 1i85g%g gd LiH ! il So fii iE [f1 lelegr-ssesacas| | | iP31ig fleas | agasees oiss} [IH zt i: : Hizipdiiiiit ind iE i135 2 2 aPpllE sEpaisI g3iydiE iiraziI vw eg8za3O Iii8ziC speesJtiiggl flispsjsiiiigalssss liippisiiiiil|ssis Ghd 2 _-- mm-w-M r-rm ---- m-- m a Project Number 454-109 s |] 32 25 S EEE A4 I i 2 C io n TE, 8 fil Li 3 3 337 I i<H= 2g 5% fe ig E5S5})| EEE RARi Te <if 33 0 jee I 323& Bi 8 s 3 82 8un8za9ma8ww rR 2 8 Gulag oe ----------------------T-------- ---------------- i Project Number 454-109 RE : gi EE i:y ' i Hi Ts t 3 iBil :! HH i: i! iisri) 3 7 FF iiilf i Ghdola - a Project Number 454-109 HE 3 Helo 5 i3 i 2 EH pd EIa | 5 Th o uv za EI eee 3 2 3 3fF 1 2 iBE IE i:$a ! H F3 E :o0el) es |: SE HN ygof HE i5i i. ii: i il 1P3E3L0 1S.C3C5]4 siit | 23 ents I as Wildlife International, Ltd. Project Number 434-109 Appendix Cerificate of Analysis AN CSentare miaLtaboeratrorines.sIn.c. CIonNvTe EARnIMyCML ErRimTiTGInor nRFROIeFOnACeNAA:LYT0S1IE8SA ome R frre wiees e Mem }3 fm Em b 3 iFaiy m cne tip Eee one & smamny [ro] [RendhoaTsSoheem TIGR)| ~~ | a = R acr 3BPre E)mN }}3 Te fmen i conan L|PPOoR om Rrmemvmv emcsi||| mem | ence Na ome Dei ioctede| i& oS a Soem mmm 5 Ty i\[3dohamimmnmmsy bhiwee antn 3 tmny rats on1a? I wie Wildlife International, Ltd. Project Nomber 454-105 Apepeen?dix Cerifieste ofAnsyss Centre Analytical Laboratories, Inc. o an: yn CoInNeTAEsRItM CLEaRbToIreeFrIisCCAnTOEROFtAaNsA:LY2S3I0S10 DuttAnyi. 3100 [-- SoneCrins: Frm $10 [-- JpEiimitaym=1o0KtncotT oinl o o,oAelo 5%mAtLp CS3H07pEKut,H$1 Y Te en "rosin pci i fr Bert ried rSyaby DmSigsmlystspilt miaowpy No doerwsn frSmietbeimrheerlssedp orir seeo l1e5ata0yb bndlheae dtercryaed rwoiietnngb0.y Bd potablfere TTPAhy TIA TRRrrifuoroEeoseetuiIscacitd,s he ile irs dn heepte fmin,FSO OW=53) iwk soc EAGndLabrieyrc Sundr (CER 160) Grn JE ae Wildlife International, Ltd. -------------- PeMumberastls ApTpeonedix CertioffiAncaalytsies a ate ee ro CoIuNrTAERvIMiCLEaRboTrIeFiICnASsOTNEROFtANeALY8SIAS [Ev-- tT mmm ~~"wwe em eeer} rm oo Tw 1 Crem -- Nt:Th 41 isivwireSbeodtiiagteCCtoddCCcasuttalecbpitns,, rm ie Pry Lafm lr ER i , araein Review Ghostee ]Seero= h common f- Cale -------------------------------------------------------- Wil-- dlife Internati.onal, Ltd. Lae Project Number 454-109 Diet and SuAppplpemeenndt Formulations Wildife IntemationaTla,bLlde. Game Bird Ration' oREDIENTS FWiShnooeyBaeCaoinmdiess, 48% rin APArTwoanyBSMpeoeci2,00%P4ePrnin DrCreodwdWhLeiymestone V[V iehrani iiannedrim niesrt+ PLirqeuimd ieow) GSToLoFullerome(dFermatco)' PERCENT 00) uSnGne5s0 GSooa G3255%0% o0t3 05 u00i25 VSiarmiin aand Mlinergal Praeims,owhoenoined 1 0.32%ofthe in, VViiaanmiinnDA, NKoinan PaVmianinBrAcid Foocnhed PTVhiiaaommiinnnee VViinnmiKe(endions Dimeiypyimiion! Bis pCiopper oSonnoeom fr "The guaranteed analysis is a minimum of 27% protein, a minimumof ? Fermentation By-Products (Sourceof Unidentified Growth Factors). Amount Supplied Pr Ton: 2T0o0o0m00ia0c1mu.s. sToompe oprmdee 20001102138aamms 1025 ppoamnss fSU5 pmrsm 2 sigam. 2.5% crude fat and a maximum of $% crude ght ..Wildlife International, Ltd. Project Number 454-109 Dict and SuAppppleemnednitx FIormations Vitamins and ElTeacbtlroelytes Concenrte GUARANTEED ANALYSIS ViVViinaaimninnngaD j&Roaaytnonh Acid FVMoSieienAci1d 2 TRPhdaioneneeaCyatdochoride Water Sas Powder Pardon 2T0000t00o0o0o Z1i%3o% Lo2&ns a550 Perl, Ts0o00g00o1cu S0for0ve0on1s 1s0gu00om0mgyM LL2oo6oo0b0sys 15000 VVIiiNtaGamRmiEinDnA)I.ESNuRpTpbSla:efmevnitn,DS-uApcltmieven,Andim-aClaiioPaserctof iVciinnSDuy)p,pAllpeh,a TVociopihe1y2AcSpeplGeomureene of iCNeonraedioenaeAncSosDdiSeuovmdoBmiCuleo(e,ntCeaofliaaChmrK)., FaalesAch. TSaumtion.gF1r1i AFymidioine yCotae,hP,si TCMhIeXveIiNaGmirPntRoaOnwodCcEaDnoUnRpEaee:)milwosrroetpwrmeedra8 ationofsprsim2tglramysofDue iin and Godan r--r ---------------------------------- Wildlife International, Ltd, s0- Project Number 454-109 DiAcptpPernedpiuxraton 27,200P1;reAmpriinle1f2o,r2P00R1O;SMwayee3 parnedpa2r3e,d20o0n1;JaannudarJyun2e,4,20200011.FeNbormuairnyal6parnedpar12a,ti2o0n0w1;asaarscfholl2o,ws6: and Control: Nopremix required. 10ppmai: 2563) g PROS + 097.4 gion SOppmaii 1281658 PFOS + 8087.2 ion 1SOppmai: 384494 PROS + 8061.6 gotion Wpomai 51266 PROS + 809gr4ati9on sweetigashiedBdeansraadtiaoetntiwosnad,ss5.p0l0a0Tc.he0ede,intw3assWuabwsretiiangngcheedwbalisentnwoder:earTgdheiHhotobsaserstrudembisdxainwncegeigbwhoawlbs.osr.aTshAfepoprmreeomdaxiionmianttghdeyraWt1ai0or0niwnoegfs "blleTenhnddeberlr.enadnTedhretwhleaewnedriiegnrshecdbonottsehnewtesasmweerrisensmewdiilchdeseatptparssoidxeiwmiraatcthilosyntownaisetihmdetihrmeieoinnn,edibtrehannhgsfaeidedsdeed0b1ehi0n1gmaibdndoienwdg b1oTwnhlee oorefmmhaeeinnpisnrgwiessreewmaeisxirdeewderpiargihooexndi.mwpaaltsaecaledyddien1d5smptoiprntoehepsr.bsocwyl,nlaohbeeulesbddopwlilamstwmiaecsdaigptell,aycwedaefitoegnrhmeaidxH,ionbnga,drt2s0tm0oi0rx.ee0dr.farloazfneqnd.utthse As needed, the propriate prem was incorporated it the Fal diet as Follow: Opmaic 5225 kgraions 2750 ke limestone W0pmais 2000 Promin + 025 ke ation + 27kgl5imest0one SOppmai: 2000 g Prem + 5025 kg ration + 2750 kg mestone 1S0ppmai: 200g Premix + 025 kg ron + 2750 ke limestone ppmai 2000 Pemix 1150.25kgration 2750 kg limestone he diets wer mined for approimtely 20 mines ins Patrson-Kelly Tin Shel Blender Wildlife International, Ltd. "sie Project Number454-109 Appendix IV "The Analysis of PFOS in Avian Diet Canal Wildlife International, Ltd. _~52 Project Number 434-109 APPENDIX IV ANALYTICAL METHODS AND RESULTS INSTRUMENT. Table1 Typical HPLC/MS/MS Operational Parameters * HCohwrloemtattPoagcrkaaprhdwithMoadePlerkin1-1E0l0mer HAiPgIh 300P0erMfaorsmsanScpeectroLmieqtueird ienqmuuilptpiepdlewiotnh raeaPcetrikoinnm-rEiltmeorriTnugr(boMlRoMnS)prmaodye.ion source. Operated ANALYTICAL COLUMN: GUARD COLUMN: OVEN TEMPERATURE: Ksiezyes)tone Betasil Cy column (50 mm x 2 mm 1D. Jum particle Keystone Javelin Cyscolumn (20mum x 2mn 1.0) 40C STOP TIME: 5.00 minutes FLOW RATE MOBILE PHASE. 250 pminute 70.0% Methanol 30.0% NANOpure Water containing 0.1% Formic Acid INJECTION VOLUME: 1004 PFOS RETENTION TIME: Approximately 4.0 minutes PFOS MONITORED MASS: 499.0 ama 99.1 amu cnLai _--_-- Wildlife International, Ltd. _-%- Project Number 454-109 APPENDIX IV Table 2 Calculations. "Theconcentrationof PFOS foundat theinstrumentwas determined using the following equation: PROS (mg t2/1)aat instrument _peakareal- e(vintreept) Determination of Sample Residues (PFOS) equation:`The concentration, expressed as ppm a.i., for cach sample was determined using the following mai in sample PFOS (m8 2/1) at instru. PFOS (ppr ) ple extrac inal volume (1) x dilution factor initial weight (Kg) Determination ofLimit ofQuantitation (LOQ) "The method LOQ, expressed as ppm .i, was determined using th following equation LOQ (ppm .i) = lowest standard concentration (mg 2.1.) x overall dilution factorofmatrix blank ovenoveralddiiluion factor ofmtr blak sample _=extraScSE ina vonleC(u1o)mxuedtieluntionffacotror Fortification Recoveries "The ppm ai measure in cach sample s divided by the nominal concentrationofeach sample (forified eve, ppm.) This ratio times 100 is the percent recoveryofthe method at tat eve of fortification 6Recover=y_BoEma meaesiudnsrameplde 100 Gntan EE ---- 2 i I: 2 z g g i i 525 sex .238 228 23 13 1] 523s goce faze me | | += = oi 3 115) 3) szes sam szam s3ae i |g FEE 2 2 3 3 |j|| S3c20o28o22 s2Ee4Es2Es2 c12c82c2do2 BcEiosaaef| ef i 1337 BEE IIE MBER |: if of Hi all all lll E 3 55 $55 555 i 5 i 838 3 Ils 3) | Biss Baer Bais Bit |e 21 78 Taal Cas| gE (HEEEREEEE OR ERA] i GRLANL --_--_--m -55- } 32 i :: 3 g 2 sg 5 2 H A Hi `g82 2.Zz2 88 58 i < 2 3 i % 213] % 2 sil 5= %IS 85]|f. tgii$ Z|8 g| 3) |g 2 5| * i esq seg 5g ssp | i3l 8 SeR LR can 25 i a LE BE EEE one 29% ve gro | 25 g55 g9 =< sss 3 B10 330 11 gd [Tas E$tGSEef EPfCSEHE 1|%HE3 fi2s3a3 da23z3 3a3 H3E33 | tif i cana? _-- . z i 2 i ie] 2 8 8 Z 2 z & 58 MSii2ly5o oBRf: 5 H dihe: biifi % HigHs. z ERE 75 I ul ess 5 GHo| E53H383E2 5H333E98 HcanEgiiss H+IE] emFirars umessems _-- 5 g i:: iLi. . 3 z BO SssBszEzzesiss 58 tll afih|353 2E%3: ) ii3t] 8S85838885858% HS I LLB] mvmmen Hil scansse 3 2582 ccgsmarseeseesnnzasznmzaosgy conmommnsieend g sevsensrcosues}iE I iz 5 A GAL? _-- s- g 3 bi z g5 3i < 3 Bl 5828 558838 28 i ik pFE lEE aIEoaEoE a iil. zg 8s fit #357 #3dacd 8% 2i3E88 I 5i) .2 $+5158| evua mmivan ae iii- a os % enting EE "59. 32 5ie... 3 i A JHE 55 @ = Il 1HMEggEEE 52 aTe E2 5 . $l] (Hy =on oem ee FsdlE 3 BI sos oe g i 3 E 2s Ee i ERIEHERE 27 3 iis: E - o = 2 |iF: Hi foe 3 it Ka i 5 t5g 153 Eel >= = 2 i EE -- ont I Wildlife International, Ltd. _~%" Project Number 454-109 APPENDIX IV. METHOD OUTLINE FOR THE ANALYSIS OF PFOS IN AVIAN DIET | Prepare matrix fortification samples in the desired avian feed stock using the dry mix technique. + | |"[ Weigh 10. sampleesoaftnhestmo8fa-0te2i. bFlranekn,chmastqruiaxrefogrltaisfsicbatoitons.andRteecstorsdamwpeliegshtnsto weigh boats and + For cach sample, measure 100 mLofmethanol with graduated cylinder and transfer nto th French square bottle. | + (Cap bots and place on shaker table.apAprlolxoimtahteelsyam2p5l0esrptmo.shake for a minimumof 30 minutes at | | Vac er | + caine ie pape an vs ein od 3 ies with ethno ce | + | || "Transfer the filtrate into a 200-mL . volumetric flask and bring to volume with methanol, { | | HPPLreOpMarSeIaMpapSronmperti,haotdewodleiolgu0yti,0onm(Usse)etaomenbtrhiangno5fli0n%faolrcNionnAtceernNmterOdaitaiStoenwdaiintlteuotritohdnesi,caiolfinrberqasutioiroennd..ranFgoerofaltlhfeinal | 4 Ampualndasutbmeit sample for HPLCIMS/MS analysis a Figure 1. Analytical method flow chart for the analysisof PFOS in avian diet. Gans -_ Wildlife International, Ltd." Project Number 454-108 APPENDIX IV (FToeneTs Ecsotvaat1.S10 a BSoreblSiaaoSowers Pe 12000 || 7 ]] oson| | 1 o a.m 5.00 1.30 ' 0.0 Figur2e. CTypiucailsrwcealivigbhrtaeteidon(1c5u)r.ve for PFOS. Slope = 11487727; Intercept = 883.10; 1 = 0.99974 GL ee -- Wildlife International, Ltd. ~~ Project Number 454-109 APPENDIX IV sTiooSno SOG mg ah We 7201 17S | >dation Hors Sartrs a. : 1000 5 | vee wo i1: 045701 23 Neh 50 P i MCaawnatro. 310 , BMulel. Wwidainh 355 8 | | RfSTorWonnefeecs) 21 0 5p 1 Haog s2117518 a SeEtarnt gTiime. Win w0a5nw0 2. 0 i mA fie He |i Tee AB 06 13% 200 200 AF 4M 471Time Figure3. Typical ion chromatogram of owPFOS calibration standard, 0.00100 mg iL. (Monitoredmass 499.0 -- 99.1). end ES Wildlife International, Ltd. ~*~ Tecjots Mostar dst100 TS SoeSOO min roe ara -- UsaAves lndooiaa vens | tGnm s1o6 MnhwB.aWiideethn :3530 Son EatxopaecteAadTaosrs402 SSaarTire iam eegratlonTvypien 4-0:838 APPENDIIXV wed Frr,ar war . 1o0 w ol sy 000 240 " he | x" p2weaz 1s we 2 Ge rthegTe Yh UH AN WEE Figure4. T(ypiMcal oionmctahssro4o55mr.0ate9o9fg.d1r)h.aigmh PFOS calibration standard,0.0100 mag/L. Gatos Wildlife International, Ltd. "%" Project Number454-109 APPENDITXV J ossr wea Wed Fert, or 0 dsm pois || oamenssuas re many 0000 | L1: i7nton ms . NvlSowasaivTenmasm. 351.00 k.l wn 3 BSeEToaWmne(sracs) 20 ia .B a ve 0 | FESantgoiTinnme en 0a00000 =p Th Eorn Tn $ h te fe Uoe Op WOE FigurSe. Tiyndpiiccaatles itohne acphprroomxaimtaotgerarmeotefntioan tmatiroifxmPbFleOaSn.k sample (454-109-MAB-3), The arrow (Monmaisst49o9.0r--9e9.d1). Chas Wildlife International, Ltd. 5 Project Number 454-109 APPENDIIXV sTcRiEoEn on. WASH Wed rn 20 1065 438101pen ama Sinirs Use brea [1534r058e591 729 M avitav 310 a6 BaSlsoai.oWnooh) 3250 ExavpgecnedAaTsreast 402 ESaxntTnine a@ avn hgdathe 42,5 [I 0) ery: = 3wo so Ho ` pw " JURTE wTw TSTnTI E T TtEh Sh aMhG En HR Figure 6. Typical ion chromatogram ofa matrix forification sample (454-109-MA5S, 50.0ppm aii) (Monitoredmass 499.0 -- 99.1). GLa? Wildlife International, Ltd. ~%- Project Number454-109 APPENDIIXV JEaHc WE WedFe0v1 ,1G wsAsm puis TUonks Suv , ners: 100ep. i1N:s4s5oo7iw2on 85s0 miwnaavmenne 3610 Sleiann) 3260 Sean EpheacedRST552 402 EOE Height 5213. ESte xartTTimg e.Wo 3i05850 wg Toe A-:68 . ., I" 5 "a 2,de7. presI" RLa we te fe Yh TW WER Figure 7. TDyipluitciaolniaocntcohrr=om2a0t0og0r.amofan avian dict sampleon Day 0 (454-109-42, 10.0ppma.) (Monitored mass 499.0 99.1). cn1ag "a Project Number 454-109 Appendix V Diagram of Test Layout from a Mallard Reproduction Study with PFOS Room 56 | wha| wn EA 150 ppm a. Pens 261 - 280 50 ppm a. Pens 241 - 260 ax = win Pens. | aie Pens. | he| Pens. aPeonsa. | ms Pens. | Ae| Peas | Aepens 10 ppm ai. Pens 221-240 0 ppm ai. Pens 201 - 220 B 271 E 2 RR a| mm| A ------{ = approx. Im = 6" gutter a = Safeguard Products Inc. Model No. 5355 b = Four X 4 ft. Chroma 50 light bulbs Note: dsPensidor lod spl. 217-220, 237-240, 257-260 and 277-280were used to house carn Wildlife International, Ltd. ~68- ProjectNumber 454-109 Appendix VI Reproductive Parameters I EggsLaid Definition - The number ofges produced durin the breeding season. S`eenpsriotdiuvcittiyve-imTphaiismeinst.a pItariamaeltseorhitghhalty sisensfirteiqvueentotltyheagfefncetreadl cboyndicthieomniscuanldserthwahticchautshee {deissttuirsbacnocneds,ucette.d, and may be adversely affcied by improper diet, crowding, excessive 2 Bae Cracked DFeifnienictriaocnks- cEagngnsotdbeetedremtiencctdedtwoithhaovuetcwrialcikzeidngsahelclasnwihnegnlianmspp,ecatnedd iwfiutnhdeateccatneddl,inwgillaimsp.s the datby adversely affecting embryo developmen. Stoehvniescirtkcinrveiostwysd,ina-gn,Tdhnisfdepwpaercackmhieentmgeircbayilssthnehobaivfrerdesqcucaeaunnstlealydsoacfifgnegccrteesadhseealsegctgrhaicncrnkaiicnkngig.ngi.s dIimrpertolpyerrelpateend tdoessihgenl,l 3 EgesSa mDToeerfcihpnarinetisiceoannltli-yngAdlhalimsacggpegasdraepmglegaiscr,edaisnud(n0dtechrossaeibnscieuhlbeacattiebodna,soerio.c.f,gretfosetharelelnlceetghgoiscrklnhaeiseds fmaonilanoluwysisinsgc.,rapTcahkreeadmreeteageseros,nvible embryos. 4. Visble Embryos DTehfiisni6tiodnet-erEmgiensedinbwyhicacnhdifientglztahetieogngsha13o-c1c4urdraeyds aanfderemtbhreyoinniitciatdieonveolfopimnceunbtathiaons. begItuni.s difficultto distinguish betwen infty and an carly embryonic death. SCeanussietdivibtyy a-nThuinsfaivsoraabflreequeennvtilryonafmfeencttedforpafreatmleitzeartiwointhinintfheertiolviityducotr,emibmrpoytoennitc mmaolreasl,itoyr chemical esidue in the ces. cnn Wildlife International, Ltd. .- Project Number 454-109 ReprodAupcpteinvdeiPxarVaImeters Fage2 s. Live Three-Week Embryos Definition - These arc embryos that are developing normally after three weeks of incubation. `This is determined by candling the eggs. `Sensitivity This parameter is skdom afte 6 Hachings D27afii2i8oonf-iEnmcubbraytioosn.that mur,pi he shell, and berate themselves from her eggs on Day Sceonnsdiittiivointsy of-inTchuibsatsioan,fsruoqeuheanstlTyetaefafencdtaedngplaeroafmerotteartiownh,ihcuhmiidsitayl,oanhdigthelympseenrsiati.ve to the ~ Humidity: 1100 ry, ducklings will ick o thie sels ~ TempSernaetsureh:rch1y0n0thohtaavnidnhtuhmeimdi,ad-iocgkginmgos bwiill gnercoewss1a0r0yfstpiapndthbeer tsohoelllsarge for thelr 7. Body Weight - Htchlngs Dupeolninrteomnov-alThferoamvetrhagheacwheeirg.ht Hoaftehhalticnhglsinrgos mbyapcahrewnteaelklpyenlotofofgorisgin,rteakweenigihmemde.diaMteealny weigh foreach week and mean weightby pn are repre. LSeonnsiitfivfiteyt- Tfhriosm icshaemioccaclasrieosniadluley afhfeectSe2d8parameter snd may reflect some residualo latent 8 14-Day Old Surivors Dufinition - Birds ha surive brooding for two weeks following hich c`Soenndsiittiivointsy u-ndTehriswhiiscah tsheldboirmdsafwfeercetederpeardamientaerdarnd birsopordorbsabtlhynmtorhechienmdiiccaatliv1e0 owfhitchhe Te dls were posed. on1ant ------------------------------------------------------ "70. Wildlife International, Ltd. Projct Number454-109 Appendix VI [-- Page3 9. Body Weight 14-Day OW Survivors tDceaafkciehnniwpteieooknly-rTelomhtoeovaaflvcearrgasogmearbtohwdeybirgowhoeeiddge.hrtMuoieftantahtewe1i14g4dh-atdyasfyoorofcladagcesh.urwveFievokourtsacnbdny-mtpeaaerynenowtdaelisgpuhertvnibvoyofrpsrnifnoa,me reponed nSieznedsi - This parameter is seldom affected, and more closely reflects the rearing praies 10. Ege Shel Thickness bmDeeefanisnuiortpeieomnneend-t,TshwaefosrhtehGdia,cchkancnedsgdormfieteadhseufrosrehedal,tleaansddtptoehnenmmweearknssbrtaarrsnorofmetphteeomcpegratautress. cqEugagtsohrelfleirchkanvicnng IcSaenrnccsalirlveyimyteo-ncTsrhusirusfraeicsea,acacsunerdladccoy.mliafPofeocdtseepsdoespiatrmsaemcoehntaetnrie;camhloEwreRvrEerrE,RmnteeshSnsdueeremme.enmtbsrmaunset cbuerlainkgea vetrhye nian _-- ES Wildlife International, Ltd. Project Number454-109 Appendix VI Replicate enificaion ExpGrrmouepnsl RNeopmhboetre__ReNpoemibPeern RawNuDmabaerpep 2Male2 BandFNe8 ummalbeer Control : oppmai 2 3 s s ` 7 wm 0 wm aw w 02 wm wm m wm mam 0 204 ns a 0s 20s wo om 206 206 EI w wn woe : 85 22008 20098 3u4s3 a344 1 210 20 wos i am nm was 2 2 0 wo am n an an wm am 4 214 214 us ase 1s 2s us wo as 1 216 216 ww naan _-- Wildlife International, Ltd. -n- Projct Number454-109 Append Vi Replisste dntifcation ExpGerroiumpent RNeupmhbeetre ReNpoombePren RowNuDmabtearpon D Male Bas ndFNeummalbeer Treament | an 2 ww 10ppmai 2 as m mo om 3 2 m mm s 20 24 ws we s 2 2s mom s = 26 mw 7 m = wom 5 24 a wo aw 5 2s 2 ws ase 10 26 250 wo am n m nm ww 2 2 = om n = wm aw 1s 20 24 ws a is m 2s wae 16 m 26 Ww aw ohaana --_ a Project Number 454-109 il THEAHRRIATNEIZARSD Eo 8 Zg 3 5 i53 833 $ i 5FS a io i claxgagsera 2o388g 68 F-3R |E2I3E8E8I8c4Eg8EugE3LELIEEEERELE LEEEED i gRavggesreTanI 78 |S3C [REREREcEeRpRaRRsRgEsEsReRsIRTz)e | 3 i FESIATIGIRCOAIRN f 1|5 l|HaEssansapzsnzdEzdzzesssss|ac|}i annsesanane sass as | 3 i i PPPPPPTETTYTELILER is i g|ssszzzaaaaaaazas |e| 3I3H GnLant ----------- J-- -T4- hl asareranaeannare a Pols 33 3=laRzEsRzReIaRaAnRaAnIQzRgSaRgns|88s2 : j | uEEgEERiEEENaEy Ef - % i fi reerernnsovaanan as 32cfLIs3 ].[: | lRsEzSsEzRzEeEaRnEaEssRsEsIsEsRsE|IgAs| 3 i BS x 8 8% i sasesagsavsuceacsusmeeasf|aans|l } |. 3 1 Ii i i i 388g fEEeLFER3 8" 33 18f%y] esausnnsraecnennannansseznssrag|-s| B3] 3 2 GALAN J 5 Project Number454-109 ii "FygetERATEANEE a Polssil sagzaaneigaaasss |g HEARE EEEEEREEEEEEEE 33 biSloSsEeRsBsRpTeRaRTsIsIgRgTsngOFs | as "a is Jil eoaserrannenenes 2RENBFEE3 E |saszrE azesaE szsazs|as| I 8235 : $i vg o8o 2-3 %i % 333 iHoaaaRgrsnreancaziana sa i2 [|1: |FeEaEzazeaeaezss8n1e3zEa8z5aEy 53| is1 heey | i eTSlasazzasges-ana ds | se 33 si lF esR sszE assE annR zazas|a| iiHf Il i wAaTpRaRsRaArTcOzIaIgGaRnennls| s| 3121 s |R sraaE zzEzsLsE zEsEaeL asg|Eael| B3t |I], | 3 Cn1Aany - J Proj Number 454-109 jj EasaveannreiaaaT| Eo3i Bj aaneannsnanses| ar .i3 i RRTRGISR-FIRARG |S ' > 8: 1 % FEC s * i gmagigeveTeaasan ag 23 i = i: i i GYRACRRATETATTAN AR HH i fife an i i3 ol 3! essesemensn HR i CALARS - n- Projct Number454109 gol aE i a i Fg . i 3 2 o 38 CBM wd = :8 |r) Ii ol. l|zp l|EsReEsRsRzRsRsEzEss-sesss|sa| ii . 3 ilessssazss assess] |FRgREREIRRREREER gg) 3 fp) ssaedgresarenan an is LE 1 33 i | saEAERRGEARRAERIARAETRAENNAALE|ER| 2Bsis: = 3 i07y|ssessszeaanassva ac sit 34s 3 Dolsssssszzagiznass lan Hi ii: = i SSXRCANARAARA ERE 3: P| AEARRARRENRAARE 2a| SEY cras=n ee -- i. ctNumber 454-109 Eo fk a i Esl 1 33 i ii LE [Bela s |FlRegRsEzArZzEgRnFzIeNsReEsEoCgAsEEfe) i laornesesasngaavs as i- i LI 3 i i |eessrarezszsessr (ic) } islsFaAsFsRgFsFsRsIsRcLaIRsOeIzRsNs | s; s | I 3 ii 5PO|EazTssszsasassesg:s|ie| H 2,3 3 fiHlsaersespsnraennnzeanannzgeesny)s len |i3i35 GNLAGO - -79- Projct Number 454-109 Eo 5 ii SER CBR nw 2 = i 8 i5 g5 : 33 |eIhave nnn pe i3 z 31 i EEEEREERE EAEEEE] i i i5 iS| ga2zgzgagsEeFrasEzEsgeRsEsaasaey 4d2g HSZ. i 3Slessszanszsanaass 5% | 233et BH i-- 32 s|BxEoRcEsEaRsEzEnEeRRsEgEeIaEzEeI|RsRgY | BIi4t% fis | 2st 2:8 i s2e8sasnrasseEnsaEnaaassaEsEassE |lfee H5H7 GNLanT [ES . 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Project Nur454109 g| EERE ee HEEEREEEEEEEERERINER 3 slege5330 88REaN | 85 bE] Grol Lis : EAE = nnn (7[2RREEEIRRERAREL slgseagyansgzaassay EI EEEEEEEEAEEEEEE ER 2908 Ea Bs | 3T 3 (oliiinrescazseszag ie} "lAaaazsaRsIssIsIsAaRazRaIsALzEEyA 25 2|g "laze RAsEiRARIRAEE| ER) i | "a i enLang -- 0 Pot Nome 454-109 2i A: do alo Lge op) 8 E53 3 35 s 33 | 8| 2 2 JN IEE o3e leosszzrsscegrsza|ze| olReEgSaFrIsRsEsRsSeAsRsEzSaEcIaLs| 83] g5i8 "lesaassggsgsszssg 30 fi |ESET -leeBnEEzREgEsEaaAzEasREsRsERaAsnEzSsas|aREnL |3ii |elsesszarazzeazezs aw nl id } lsrsgraeggsrageeg|gn|iz LAA Ra - - 33 | sf srt - _--_--_-- - Project Number 454-109 3 al ygoall |" ' CAI ela] FxRaEg a# l ce 3 FR EEA SESAE IEA 22 sl aE131 42 TlEeEeEAERECREcAssEsAgNeEsD| | bEeF)i , i classsags saRaEIER ES HE clozgrreasarusgrag | RR ii: i ~lssgsassseresgang | ia 131 lesesssssnesnasselie gaf "n1anG FE -- tte PctNore 454109 zl ox zl ox zx a= Bla ckks aloe sooo alo kx ElER =p: co rere 13050 gfg e |: e Elene 2 ee isiff Hl ele ow 33353| [a]= slo x tBeEEilxsR| la. ox Ble ror om meni]3 $5518 al=. Blo ox ex mE E3E323E8 22 | fal oo | lelelp fe) lel eke i 2g glx glee oe a= loo ke |E . 5 IEZz IioT: o,g F 3Fo 3 [3 s| 1, Bie E40 fF pig) | HleRasisiiainii failing! iig:8 fpr LEZESEEES_SEREoccii(2 ean? ------------------------------------------ ae rj Number 54109 al ox- gx glx gl x Bx glx: gf | Jalon allx ex glx 0 mx Bloex 0 wow lx Alo x amt shoo 0 axe mle oo 253 aw. gluse. oo slo l=. 0 HANH HE I 3 2 irr.ed 2isf | E (EnEee2d2734: 0NLang _-- 87. Project Number 454-109 Appendix X - Table | Adult Liver Weights (g) from a Mallard Reproduction Study with PFOS Control (0 ppm a.i.) Male Female Pen Liver Liver 201 31081 36870 202 32.528 36336 203 33915 46728 204 25209 45.564 208 33438 57.924 206 28829 46.002 207 28.689 8662 208 270% 59.838 209 25919 52441 210 - 21.509 21 27.129 45.440 21 3009 47038 2m 38599 37995 214 27.888 25939 21s 26030 34229 216 24.921 33.845 Mean 2.422 41648 sD 3864 10512 - Data rt reported due to mortality. 601s e ----e ------------------------------------------------ se Project Number 454-109 Appendix X - Table 2 Adult Liver Weights (8) from a Mallard Rep1r0opdupcmtaioin Study with PFOS r--r------Male-------- Female---- Pn Liver Liver mm 30u66m3 s36u4n5s4 mne 7a0nms ssusean ns F 2m 25830 SPr6e6e0 mns 2552300 asomns 2 0 a7 525) wnemn mni 3360867006 w30a41e9 mne S02986766 s2.1u26 ns 01 26 35314 263 2m Mean 0I6r01 2r0e7 _ T Toe shore ifeces fom he ool gope werentssasigan Soden cs). CALs Wil_d-- life Internati.onal, Ltd. "89- Project Number 454-109 Appendin Xi Histopathology Report GAL a A -- EPL' Project Number 454-109 STUDY NUMBER 454-109 ' EPL PROJECT NUMBER 212-028 PFOS: SR A REPRODUCTION STUDY Epa ag SubmittedLby:apa pA -- ig-- h Submitted to: December 27, 2001 Gada C--e -------------------------------------------------------- EPL' " TABLE OF CONTENTS sna a . Sr " nL _--_--mm------ -92- Project Number 454-109 PATHOLOGY SUMMARY NLA L _--_-- EPL' ee 110 ~ ours seat `WILDLIFE INTERNATIONAL, LTD. PFOS: A REPRODUCTION STUDY WITH THE MALLARD PATHOLOGY SUMMARY : nr war Light microscopic examination oswas on performoend are sd sections of selected were untreated or which received 10 ppma.i. of the test article (Perfluorooctane Sulfonic Acid, Potassium Salt [PFOS)) in the feed for approximately 21 weeks. en hn (Ae Ie 10 evaluate the effects upon adult mallards (Anas platyrhynchos) of dietary design was as follows: E Tm E Experimental Grovp|I Taos |Farmal-- es |-- WalesT].TF-- orres | |"Conolppm) | 20 1 20 1 3 | 7 1 L__toppmai | 20 2 | 3|7 | "Offspring did not receive the test substance in the diet. All offspring were euthanized on arenas pusrions age. Necropsies were performed on all birds by Wildife International, Ltd. GAL --_--_----_ "%4- EPL' EXPERIMENTAL PATHOLOGY LABORATORIES, INC. Projct Number 454-109 Waste aman, Ld. Stuy Number 454109 Tissues were collected by Willie Intemational, Ld. from adult and offspring, fixed n 10% neutral buffered formalin, and sent to Experimental Pathology Laboratories, Inc (EPL), where they were trimmed, embedded in paraffin, and made into hematoryin and eosin-stained sections on glass microscope sides. `The following tissues from adult and offspring mallards were examinbeyd ight microscopy as availabe: ver, gallbladder, proventriculus, Kidnoy, brain, gonad (ovary or tests), bursa of Fabricius, and adiposs tissus. Alltssues required by protocol are presented in the Histopathology Incidence Tables. Microscopic findings for each issus examined from each mallard ar listed in the Histopathology Incidence Tables. Mictoscopio changes wore gradad one to five depending on severly. Nongradable findings (inducing autolysis such that a diagnostic interpretation could not be rendered) are sted as present (P). Al findings forall animals are summarized by sox, age group, and treatment group i the Summary Incidsnce Tables, together with the total number of animals in each group or which tissues were examined. A tabulation of gross observations noted at necropsy with the corresponding microscopic change, f propriate, i presented inthe Correlation of Gross and Microscopic Findings tables. The observations in these tables wer transcribed from the Gross Necropsy racords provided by Wildif Intemational, Le. RESULTS Except for one control (3235) adult male, all adults and offspring survived 10 the endof their respective study intervals. No effects considered possibly velated to test article (PFOS) administration wera noted in the ve, kidney, adipose tissue, brain, and proventriculus of adut and offspring males and females, bursa of Fabricius in offspring males and females, ovaries of adult and L2. ona -- EPL' - tern rn1 oa res et on the bursa of Fabricius in adult males and females could not be determined since no samples were submitted for evaluation. The bursa of Fabricius typically ee esas observed macroscopically as being reduced in size (< 3.5 cm) when compared to other birds. Of these birds, one control a re (3225) and four treated birds (3261, eto exhibited degeneration/atrophy of the seminiferous tubules characterized by an absence of mature sperm and a decrease in the number of cell layers of the germinal epithelium. The ee ee ae 2 28 remainingtwoof seven 10 ppm a.i. males (3263, 3285) en tr ih ar wit si rch presence of numerous spermatids and mature spermatozoa in the seminiferous oo `degeneration/atrophy noted in the control and treated adult birds may simply of reduced size (with or without a corresponding microscopic rat 02 observation) of the Ne neo -------------------------- roc Nant435.109 EPL' OTN rretteesssona my ha clrhs nore truvo phase A BH Hoenn `gE abseentsortnarmrowaltuA means,veanrdtheadd ge1r4mA ionayl oelpditrheltium with decreased cell layers en rrr ery five (5) 10 ppm a.i. adult females, the ovaries were indicated macroscopically as nee S A E A PTS AT RS rrres--oo--rnm--s--o--m--e--h--ena--mer--en--. ST eco nunr ofones aetcnts tehasteemann btan fconot a5nds1p0h5am.rfo, er es avanttaeAclsromavsta.rhieoTsnsopfmaofanfesipsroianatg efyedmatclioesnnswseetrcoenocmhpiartoahrcttrearaimznebodeebcyslensdumersaoousomooecyrtes Vacuolization in the liver was characterized by the presence of vacuoles `within the hepatocellular cytoplasm which varied from, usually small, poorly cytoplasm, to variably sized, sharply demarcated, clear vacuoles in the hepatocellular cytoplasm. While special stains were not employetdo identify the nna eee eee ro Number 54109 EPL' Arse mettain ho vaccine atr morhologyvas seven wie offspring of both sexes. Although there was a slight increase in the incidence of this change in 10ppma.i. treated adult males when compared to controls (20 vs. a eat tar pratense groups. In addition, the change was not observed in treated male offspring or at semintin, Ta para i a oath meen determined, but it may have been biliary in origin. Also, the liveroftwo control `adult males and one control adult female exhibited amyloid depositionconsisting of amorphous pale eosinophilic material, which filled the hepatic sinusoids. In the one control male that died (3235), the liver parenchyma was almost arusop completely replaced 0by thre daeposvitoedwoaemydlnoid--c. --onl--ano--r a--id mee ress, Teas ota vare marty dr avomer Tn rata --"---- i. cares _-- EPL' eee 50 - rs yr ee treatment because of the low incidence, limited distribution, and relatively low. offspring mallards from control and treated groups included mononuclear cell eeAntanas eriof a ee er -- tes oe, `males and females was characterized by the presence of basophilic and/or aeos so rea e?dr hnad orsd neor minimal compression of adjacent normal glandular epithelium. The pathogenesis ee of this lesion was not determined. However, since the incidencewasnot - CRLaNG -_-- EPL' Pict amr 154.109 - Vra So sn CONCLUANSDSIUOMMNARSY radorviNeoalsnesiovofensareccorunestaiedceirfnepdhrpiorsvsg,ibllykeriesnlapat,enddtoootmnesast anrttoirculeet(nPaoFOSF)aeno i afseprionpgimnagle,s haenianmriesa,svatthesaobfeacruofancdsptilnegifenmhale1s0aonmfoetos of meiecrdoscgorpoeupcbitsharaetcuos)edsueggsestcs hstePRwOhSomaywihtahvtsa ccoroeosptonsdoirgy pst Teroduciv pha egesion,a rma shysclogaphanamenon. ny acpodotdetnotnioa,lttabensoyt-tpaorettirecnmlteiiaelnffeaecsstsitnonas trhefabruresstaeosafnwFtiarbereigcsiiuiistaindeaodseulttomvamlaetslas,ndafemales rfiercinugtcaonuydaneotrabotedfdiiavgenloys,StTohermfeiwnicvurs ochtaengeesxornoastYeloymhotf eu andor afaprig malls rm contol and 10 pm a. hterope ere considered incidental and unrelated to test article (PFOS) administration. 7 4 . AA. / EREDERCH & RUECKE Si as Shara AC Veterinary Pathologist TY [27/F03 E FAR/cp 2 GLa nt _--_--sMe)oH mmmee --_--_-- Pro Number 54.109 EPL] SERENA PATHOLOGY-LABGAATORES TG compLANGE STATEMENT oe mate mers 0 Shouts or maga cnotte oa Species _Mallard bat. tr. racer ruNumber 212-028 TSotruosd.y. A nepuycintooatns TnAdidle_Perfluorooctane Sulfonic Acid, Potassium Salt (PFOS) Th iscpatiogy pron of ns soverferc aywareconcadin compliance with the Good Laboratory Practice regulations of the Environmental Protection Agency as stipulated by 40 CFR 160. Lp el EPL Pipject Coordinator RIVEF STH i. GnLanT EEE -- EPL' - crm e -- o SHA I tnston e--y--ga--m------ nt owe _-- ny nen oe earn os on S rrT------------------ ---- T Y/Y 03 -- Giang --_--_-- "102- Priect Number456-109 ApuLT SUMMARY INCIDENCE TABLES engay -103- ProjectNumber 454-109 h5a0u.1t09Sacrifice ole ollard SUMMARY INCIDENCE TABLE conan | 1a0-pdmat OfToPn TISSUE G0 |1 | 0| ToroigfyGrants Inm fitratet . Ronoracn lea G11_ -- = [BRT GEoE. comin) Ea-- |r CoIcntorsavaOsrcgualnairsms. T TnFf etrraitev,asMocnaoneuclear Cell, | " TaoscloPaceiraici |1I -- 1 -- -- -- -- a-- -- -- -- --] jv v or rps e - Cemen _ |] --] eterno. panned) 0) |) 1 [AutolystspreciuesSagrosis | --20 170 igo. cammn |G| Go) Coasyclood tGeprmsiitimons| | 1 [infiiisntfnrTaafmtaleti.aon,mSodnGmeosrntaaaectnteiistroeCesnll,|| 11 | -- -- 1 tropa E np [CioyviioncdaDpepmoisinteidony | G[2o)| _w-| |-- [Freepoetoncetlulieromeras1T" 1 T 7 Tynpfeirtptiraaittes,,FiWriedetoIcnftleam_itorTy | ---- en 7 Ibniteena.cytetmpoiatncecfyters om | 8 Faronsclew? CalInfirate | vacoliaation, hepatocellular|12" --] [ePReO.VESoV(rT0m.RsCIiCvniIceoLn)US| 01 1 ] EPL Se-- py CnLaNs a---------- er -- ------------ ------ oe ProjectNumber 454-109 Ssine 454-109 E EFT EL. Vasculit1s/Perivasculitis SUMMARY INCIDENCE TABLE arr rT i --_ -- -- -- -- 1 rogtFawa a --ri] a1 1 ---- meer, a -- 1 Gitaatatiorn, Seminitersss "E tire y Hori GT --T--i1 Vasculitis/Perivascul itis -- TT -- -- 1 --1 + 1 Fe ---- See -- Ti-- rm -- |, = Gada, EEE -105- Project Number454-109 454.109 SUMMARY INCIDENCE TABLE I = EE poe po Gn a -------- Chemie i GI o.ES) -- --] --] fFieririavtaes,aiaFrovonuclearGell, -- -- ----1 Vasculitis/Per vasculitis _- --] [BUwRSA oaF Fa)sRicius wo.|| i Cr a e [astoAlysie s phroeu c.iouceo ms Dere agnen t | | 20) | 20 | (9) 1 iow (0. Eiswingo) | 0) |a) i a haa [LIVER (NO. EXAMINED). amyloidDeposition Ee bbe FEE Feet Hepatocellular [I Cee 1 infiltrate,MononuclearCell |17| 15 Intracytospolmasaic| |" |__Pigment, Hepatocytes | - | MononuclearCellInfiltrate 1 --_] | Vacuolization, Wepatocellular| 17 1 16 [ jovi weaved) | @o) |G| Atrophy TE1] 11 F-- -- --1 -- -- 1 -- nda? ---------------------------------- " Project Number 454-109 450-109 `SUMMARY INCIDENCE TABLE os aa EP ERR Infomation "TT ---- -- m -- DI y--e -- |+] 1 -- 1 E ann ami eT Tr Ft] L-- C1 -- -- a [ot = EE I - -- 1 1 1 -- ft - Ce-- --FE = 1 - 17 J ecm - -- I-- --] --] E-- |S 3 1 -- ---- i se) rm Fr TT -- 1 1 1 1 1 [1 = 11] nto Be -107- Project Number 454-109 ADULT HISTOPATHOLOGY INCIDENCE TABLES cate --_--mee S108 ProjctNumber454-109 HISTOPATHOLOGY INCIDENCE TABLE ue bSr45ao4ii-t1t0Si9cirnigice Ii | | | 11] Hi | || A ! Yi { h RSHE HEi HHHHl EHHHEHEHERHEEEEHHEEHHREEEERHREEEHTE [AQIPOSE TISSUE ATX] BRENNA] x xxx] " Bots Gramm I "Monon Cel Inflammation eos-- ~ I H IT I HH 4 I " i. aaanmsmimetnaocne--G--L[DC PEx] aREE ot LTnTa] Tr] SreT s-- ame | S t TT f o TT A iopi--u RCva AR FiW ETTn "bream l LAE ALE ERASE [aaaaaman al alam fayloidDegosition I ier eJ tdSrai En ason._Hosnoonuecles tial 1] He 1 - i Hens TH Nephropathy -- -- il Thi EE. - -- i---- 3 I1 AmyloidDeposition 13 tebe8. 1z 1 se tied Inlet lHnyperpflasiia, ltMrBiixaeltdeDeuc.ts Inflamatory |ye -- --e--e --T --T1 T bi 1 --_- S nr yita .astoartat t uafccrioiemass re Cell [L LLC LLza T Ld h TIn pum a EPL ExperimentalPaha Cabos EE, TEs fn. Key inone Bemarkab7le wAeko SectCion R[=IncamSpleteoA-hutoiysis [ER] --_---- C109 ProjectNumber 454-109 HISTOPATHOLOGY INCIDENCE TABLE Cs5to.109 a ane JL H E mn T | | | fT| | wisislslsislshl ls wals(slala a sla sls lala al2f2lz|2|222]2(21212(2|2}2(2|2|2/2/2:2 Mica Sie a pT tension mm------ -- ma VascultisiPerivaseulitis. i +] [ TteEC coal bi mibm l aaa al al mnfaip san! ,e 58pf e adtaess Saran ft c"eiammee nts {|e TSTHTTe oo ee ITT] vasculitisbor iascul its HH fT + -- I---- ry HH 1 f -- r "I -- T 7= je ym SE -- TT i i 1 i A -- = Fr I-- EE T = re th HH TY Tr 2 EPL Experimenal Pablo Laboratories To. ey +XL okot Rema0rki able ANekeo Sectionlis-IncoB ngleteRAekatolysts ene _-- -10- Project Number 454-109 HISTOPATHOLOGY INCIDENCE TABLE wor slats RwaoiSsacrifice Male Mallard | RRIRl | IK Sh A I1 Iq | 1 Cse HEHEHEleHiEnHEtiRHaBlHyERaeHslRlsH) CRfogreEignIfodSrEamon LD] E------------ Carrs atilroamatcei,onasoscioar Cell || 9 Te2 [Com ccioniarrgnoE ss.eoar r Ca Hh KAEC CE ar I fircaes.aWnonemcienrSL [ JAI R S HEE wc a en re see pert | fish 7TIC) BE EA CiaEtfracR ates O Gogmsis| 3AT[AR3A0SATLaTAAR]ALA L(AhAe|leATA ATIATel TALATA] C ;wo oBe iCpHoinanncier G6] 5 | Ep BE rrr 0] I Tr -- n Natmi i ali I mm TEIm lsvmee tora por sitionI --|--|-- | T J L T AA m FMesuitoces sl u,ltai hrWoet e n. min |T | T ll T btt= iintn racsa tmaiapmt feermoime | elt {1 171L ] T L 1ECU TE A]1 T|eni ts dtemioocntesite T[E TTL a Ere ExperimentaPathologyLaboratories,fn. ott st ssn cme sits Li, Some wernt Edo fy _-- am Project Number454-109 HISTOPATHOLOGY INCIDENCE TABLE ae we ae T[T HH I | [T 1| Talemations AEN | I |||| H E A` R22Ee ae RlaRae r slaldaa dat ehlnp i dai|nshl ia ila es e,Hl: AB y.ChE ri lE enNFH n | Imm ET a ta] a| er filtrate,Mononuclear Cell 12] I nd EE fefamina e snake eas GCormccppiamdOnrgssantisote,ba. 417 rr {1TC TL 1 T e C 1 e T eT 1] ! |p BDila+ete amdtiionn,SRemmineietearonusrts1 { 1 . Tm 150 r1v E1T[aH E[TIe T Th Vasculitislperivascul tis BA i im A ee EE-- he 1 --ke@ Hr I] S =H aPH -- Fe AH Hr CT 0 HHH So Fr Ir C Co T E HT TH rr THT Em pa ana EH R R0 See es EPL Key okt Eemarkatle NekoSection [-inconglete Aehutolysts enLand _-- -12- Project Number 454-109 HISTOPATHOLOGY INCIDENCE TABLE won ws100 AfdeumlatleMSaaclrliafridce Salata ahaa a -- | 1 | Ti InN | || al] (EE ! | | [1 | H azfzizE zala]H as]3)HsE ]Ea)Hs HH saHliH] HsE SHEE ANT AAT a Ee io rate, onamcteoanoncrlerCGell| A CIT I Anflamation FET f ftoooIsnsrotndrgasatvanr ises. FAR 17]TTTOC RE x A a ER] "| inahtraeatre,bHoaragtaceiearGel. rT LN OE I ] |Vasculitis/Perivasculitis EES -- 1 4] HT] SAFRIES AA aA]weptIrn p A U ie --a -- mmr ATaTAlr EaTT aAtTT i i A hEAlia mCoosfcloaanicrdedaDotee,hrmoiagnsea[ ieGn1u1rsI|x|a |1 1a1l]anI ] r alaleje miex to a bin 3 inflsAmtteon, 7T 1 1 Ty In Inflamation._Granulamatous. " 1i. Y FE Cieiromaity [mans IT HH f Oregieeade raBtioolnctn eebcir.otsnis, [| C| Eoe" [T7 EHHE J 1m1 m Ha yapfeinr.plasia, s Bile IDo uncLtstt rs||] cite senGi al {1 r FLAfA eT e C 0 1 CTTC] F[origp eunt_ermti enases e o| nBtrom i cT areer rr onnTE ie T - ns EPL Emr Dre re Key SL TL eM Remarkable Woks Section Ielncomlete Aviutolysts [atkins aries GnLadvi EE -- "se ProjectNumber454-109 HISTOPATHOLOGY INCIDENCE TABLE Ee Tete Tiara ao dT H: s | I Hl 11 Satlafal salar aalaatsfslalatalals iH 33 2 a HEHHHa HERl E ale e 2202 I m m=e -- ---- i LH+e iRhH,ari IZ7 ro EE . stints re rmmmezeat + m---- TCR hh a EEE T -- --t f 7 HH 1 --H I ee -- FR r E a EER Er reed or ----r------_-- == --FE Eerie PatoCLaoboratories tn. or L tas sme n te sSsenrimiT ssLtwos cntens _-- "nee Project Number454-109 HISTOPATHOLOGY INCIDENCE TABLE aon tals afalstalalallalsl stalls - AFdeumltteSmactraifrigce TTT] .Hh| | IH || | | 1H | | iCE HHeRleRleRleHlEn EREHa HAHaHlHa sRH5H1ES REISE TE POT HRA [itnfa rate etnonamcieGaelri | 4TTo 1 T 11 T i HH b -- E I TA LoccoiadOsrgcanuislmsa, r_ l ---- l A ttEti] Beraciar TT JI Tala sla ESET ST JLs SoRi ICa E i {S Rnaa WW l El W aE R T Talal 7 |--Butolysis PreclutesDiagnosis Jepp plplplepief an [elele) L mm e LPe e e iver [ali njinjini jaja. kl Tn Ann CE |iAonvfcloocmiae adtDiteropgnoa.snRr iiteuintosne o | on1 iLT1T [ITL |TI T UIS 1 T 1] et --- A AmyloidDeposition feed. CE 1 it senli lede----H ai [ Ienfic lsetrnattei.e SMonoCnpupp crloisnoer[ t| sl |a E a an=dH heal wa EPL Experimental Pahalogy Laboratories ine. ey Fao SEE TE soot Remarkable Neko Suction Ielscomslete Avhutolysis RR ssi COLADG _----m--m-- "nse Project Number 454-109 HISTOPATHOLOGY INCIDENCE TABLE Gane iwnonrne NI di IHH[|Hh Emid EBi-- as pePoal-- a ithelivo, Wltifocsi Tttiriatties.pfeorniavnaisccluelstitia HWiiahdhenls e iil l nhlili i3aaaheit shisilns PAPPEPOEOOoeRRN HTT) [5] e CO i m e Xn a | Ti] | | TtTr FR 7 m= || C--T--HETe ETT | fT eee Eee ee eT Ey ~ -- ------ ------ -- a EH ae = e -f =EHHEH ESE RHEE HH L-- o -- re-- ere---- ra ------_ 11i 1 CT LrTTITT I --HH i, mAe -- e Sr e en TH EE -- =o we EER TTT I to tS s Serlr e tte cin ena s ets Coram _--_-- ~1i6- Project Number 454-109 ADULT CORRELATION OF GROSS AND MICROSCOPIC FINDINGS Caran --e--e ---------------------------------------- TR ProjectNumber 454-109 I pl] | || Ww, | a7 EE gk a 3 EE g |dy : da: Cl [] S23 C5Eg] | | si[ bs LTT CALang - m _--Fs ---m wmm us Project Number454109 of] | 3 || EEE 5 5 a 3 30: | FA | gl IT 2H 341 | | i] a. H FU 5 |3 | | " ag) i 1| day || PAE = $35ed nLL00 -_-- 3 a| we PstNumba 50108 E8oRol bhslis liglae]n3lg: ERRUE EE i CRdEaEalEdgEsTke ] 42g a03q000010111i | .a.| lissdssdy | | : BolgE i dldE | 3g SH5 8EE5BEE8EBE= 8 30 | 5| | | SNLEELEE [ E TTT - COLO _-- i20- | | i ; | gl r3 4E I25S8|dEsEleEE8slE5s F2 R4E1F FER THEHEE HER HgERE58E]E; PERNA E01 ! HEH P !| | ere = 3g]: | & (gz I : S1ig | H 55E0RdB]IoE lsR ] 2 tEFREECEHaEblE ieRHyEEEEeEEEEE 8Z | t | | a $el20dle2 ! lsB8 REE8g E8iE% Project Number 454-109 z : GAaALO - -121- 3 10] | FE 44| 53 | S$ 5 5 s5 2l5sH 8s |g8gs LEER | FREE EEEE | s3 d | 2 |g | | &FRtzE2C E&R5HEE gEg E EE EE LEH SHER 3 | gl4lEd Project Number 454-109 o: Cat Project Number 454-109 S122 OFFSPRING SUMMARY INCIDENCE TABLES 6n1L0: - S- Project Number 454-109 45Oa4re.t1i0H0nalglorSdacrifice SUMMARY INCIDENCE TABLE 1w0-gapmat EInfEiratTe, Woromctear cert L[ ofo|L |1T jeT k Gi pee n) | 01 IE -- LL a -- own) 1 ENCEE _NCE --IE) 1 1 -- [AvtolysspreciosOlagosfs | 3 1 ---- iv oo. ee iner n) | 01 [infiTtrat[e, WEeronEaciearCell - ves 0. EAIEDD or [~R~aefsieepnttcerorscteeHl,onlpoiotasre,ormoe3hleesorepoGems | 2 |ETTE1 SoEevoElzation.fepatocellular| 3 -- roves RoG , ED) | 01 [nfi--Terate, froncle-- ar Gel [13- EST Go. ci) "ee mature 23 -- -- -- -- -- ---- -- -- TT ee s-- 1 -_-- -- = BA ErL cmap omen -------- TT] oe, GRALDM nae Project Number454-109 af45er4mt-ao1lr0ei9nHgalSlaacndrifice TPO TSE (16. DINED) |tofitrate. HoromcleaeColl i a Dw `SUMMARY INCIDENCE TABLE oh I conor| 0-gpmt TT =bb] ECoT m)RE IC CO -- oro --1 -- -- E E CoAE R Tr LC 1 -- -- [eKf irwaotoe.,evxmYioennoerr0u)eics| Gell |) 3 _|0) - ---- -- we | oo | 6 _ Degeneration/Mecrosis. --- [ tTateivsirtatoe,cYeoTmmieuloeuttaeraGrI.T 53 2 aVyacuoGliozataionw,T HDepa)toceT llular | 3 | or1 ] -- Temitur 7 I ROVENTRICITS (i.EEmReD) 11 Infiitrate,Wonomclear Cell 5 | 1 Re ---- === er EE = = = mm ------ TT Ere e------r------ 1-- CALL05 "use Project Number 456-109 OFFSPRING HISTOPATHOLOGY INCIDENCE TABLES cranny se ProjectNumber 454-109 HISTOPATHOLOGY INCIDENCE TABLE owe aon Sa5ha-a10n0g sacrifice Kale vations os i HH . || || | | | | i| | | hl | | HReEsEsRRhEaHn || BPEe E I mr T PL [A TH iT ai t - or is al HE far ta e a Ja aa A SC t= o Ee veSf amiaaGl] aaa] | Te ]: psaeiila TEE [reread |--Degeneration/Necrosis. .. i a Ty rT E= een Ea h fT e E ih x HHE Fe SE n e. HongnuclearCell|1[ I Iw-- e TF-- I] EH -- Cr Vase emssesssmscecas Tre " 3HH E | H t H Er f ---- r = e -- mmr E ---- e EE -- ; FrTo rB e B ee EPL mara ane FEEEEE EE Er, T agR c ss KeyteNot Remarkable Neko Section I-lncomplete A-katolysts Gating eee ere------------------------------ me `Project Number 454-109 HISTOPATHOLOGY INCIDENCE TABLE aor aon aFsOeftfm1stp0er9inmgiSeacsrifice . th HT Pi || | | I Hin | 11 Hsellsalaesilastezellasle fealtsalleallealjaetssesll |] | || relefaalofil ehalalslals]s) | | GORnGEbicT seFS amiSne xTTiu XE KX TiTm] iTE KKKEK KAALR if 67 Eis [EaDaal AuG tolysis Pa reciudes a Disamesisl Tepe] | a[aTelA e] aBHorT KiS to mafJamba IaaA ] RaA bapaaiesaI gal TE T] i EE senate. Re]I Kegatoce lular. Acute. I YP A | [ifnascial at oHno,nmanateosii r a [33l 1 sh PH OVARY S-------- F pte] mature = Pp pip pielpipl [piplplpplel 1 LF 1 iatitrae.oorotesrGel TT[A1 12 12 LT -- HE TL ee HA H]| TT Yo T3 -- SSS| r oTe : HT -- + To rr 1 ---- ---- Ii IT im; va Ect a, E[ E L es R ws EPL ey +XeNok RemarkableNNSectice Iincomplete Ankutolysis Chi ------------------------------------ Wildl_--ife Internati.onal, Ltd."se Project Number 454-109 AppendisXIA "The Analysis of FOS inAdultRed Blood Cll and Sra GALLS ------------------------------------------------ Wildlife International, Ltd. -129- Project Number 454-109 Bun yo200 Table X. Spuommmaarysof.WPfeReoOkSrin MallardRed Blood Cel Samples 10 Towl eeoyeBuen dAwTs tiog a oewss wuoowmmee whomooonnm iliaawwm admammm oonw i e ww s e awowoeam i wy oommnaemn ues amwdmmm ammammww om om Sm ees sw euhemnmes aoSmumemn dmdmmm mvmawwm ow ome -- e m--y w_nw--es awwou.w wamm Saammm ___om -- So S-- F eeemoteE mean--i--n--g-- 00 #001 wn. ford od clsand scr ore tid 0a.0e 8 wgs. ltwtheoieotf baenveilta on sh fen cass ------------~------------------------------------ "130+ Wildlife International, Ltd. Project Number454-109 Earn sty No: os086 Table XI. S10umpmpamrayo.ffoPrFWOeSekin5Mallard Red Blood Cell Sampleast Sofw CB nn iesa ST illoiim omwes o oowpnm moGeoemm SSCeeiimmlmmowsine UooIwes oGomwmeloonmwsbb SSCloiiovsme ooommmaee osohamemwmmn>bo SSCieilvmeoennwss ooommmaaa moohommme SSCeillmioommnness mooomouam ooomemoaaw>n> tise ous onan pubeus Abnue Smaaewm ss3omoo sspiaaeie vssiaeeell saaaeew smmmoeer asiiaemm oosemoree saeamlm svsmeoerw uSTmADDRsAemTIeaONe, rAumisee 2SWea iiono owIln eb iiinn dnes 430 [-- L0Q=001 ym. for rd od cladserum Spent Fae 0058 Laan Wildlife International, Ltd. Project Number454-109 ------ `Table XII. SSOupmpmmarayd.offoPrFWOeSeki'nsMallard Red Blood Cell Samples at So Cnn St ai Ame D SRivT as Tones oemwee D aDume Dmueeemmiggnn SCaimmae wowme oomrwia> mamw E swwm pin rv-- 5 "Table XI. pSoum amd.mfoorafWPreFeOykS1i0n Mallard Red Blood Cell Samples st 0 Swoe Co SmRewwT en T a wn Si tiweo oooowmm Swh-- nea o-- oomm Lr mom a me oowewo eewe uke tSeee lpum nhas suAmmeer mwhooms mamw msowm oa. aaomha.nn aw omm iwioww 838 oo ammeenn _ vsawm osoomm - owooaommnnd menwem 2ssommme oy oummaen aw m mww -%Q won SoTmoPoelnm.saSotsmen 101e=aNor Qanniefaloeweeckonwstadineofdioebconmsosniannsa 63i00rmLie 100001 pmtoed ood and seam FSorevnadle ecrdwd m0,os00i0d00astn.ehfhehseevreoodhenlowdestdelibration [-- Pgss0ctss Ghana 12 Wildlife International, Ltd. Project Number 454-109 Baye Sty No: 023086 "Table XIV. 1S0upmpmmarayd.offoPrFWOeSeikn1M0allard Red Blood Cell Samples at Sroymer cam LCT iimoame mown moToommm sSl iimmoem nwei,n ootoummsae m CCipme oln oooommne Siiimmmmioennnin oGoowmmeas SSCiiimmmeiain oooooomnm Sime os vSeae Duondwer ooommma vaSowem ooommmenaas Smvamem ooommmowaa> smamem ooommmaaa swmmmee ooommmneeans sssmmaeem omews om T amheis p smmriopeen mmimaaaeem hbse a mimmaaawwm oy mmmaaamwe wwasb smwaamema os en msmeaumem Whlree _ AGE 303 uc ston L0Q = 001 py for died cls and sum Ege Reach Poet at5n ons me Wildlife International, Ltd. Project Number 454-109 [-- Table XV. Spuommma.ryfoorfWPeFeOkSi15n Mallard Red Blood Cel Samples at STi`OSpmo.nRsomr E. CToonmBtoeree. eGtsaonmmte ExsIthrmaecretion "meAwalysts om s* oAomatee s he SCColmiammisss omomomem Goammooewnn amvaewn ee mew hohmey SSSoammmame oanmmmeae aoommnen isvmaeewm Gemma wm omen mw m oome emohme eM SSSeemmmon wooommm bowrowow masammm m oame eW SGSimmmaewssi. ooowmmnm ouwnmoeenss aadmwm m smwe ehhe SrSmmeesg woowwm ooonwewss mm aew moemm he STANDARD DEVIATION: 0000613 Dein [ 3a 0 Na-- hhaow4espckoavsedntofdchae cionncpomdainnorsse(00n9owmyie L00= 001py foe ontclssnsr vF0or va0es0cia d NOd. 0o00N0DS,unolf.tbec eofehe lwcesgbradtioin d Eres Pacszeros GALE Wildlife International, Ltd. ue Project Number454-109 [E---- Table XVI. Summary ofPROS inMallardRedBloodCell Samplesat 10 ppm a.i. for Week 15 T wr Conrnr iSe Gea ru iuve T gamR er uu oTwmese gwomen oponeee ow wowrmt Swmwba vMMEwm om awmau w rw y wwar mmewwn Twmm mmmew= ens owhw oSwems aImm m pm l= AS Sn HI Ry IE SWAS N Oua OmSn OmmSOemWerDR Smm22nm0) sm gp amm,ma o2636 "TableXVII 0 Summap ryofofrPTm FEOSRiMn MallardRedBlood Cell Samplesat wr wn rrr teee pt aen eaHmrl e l sont E my y om oomee aMmm wmee om o---- ---- L002 001 yg.fore Hodcls ern yen Rese etn C1526 Wildlife International, Ltd. 3s Project Number 454-109 age SyNo: 028066 Table XVII,aSiumfmoarWryeoefkPROS in Mallard Serum Samples at 0 pp - Guntsee apaan Aheu rpeem CCeemnmawse oomee oOommeean TeTaomem Twmehoome bosws SSGmiiemmees wmoeeoi vvOemwnaa vmveeemlm TTJammm sboeoremmm pC Cenrm mnos vooeoos lmomoeennevmemeevSmA bes ee SSChelimum.me oowraae oTomoennn em mmweeAamm eobeemea Smiewesw oaoo ommenn me e e TsaEm seowehens common sms. to. L0Q 001 pn.tote ood els nd erm Bageabesesen ---- enn Wildlife International, Led. "16+ Project Number 454109 en uy Noss "Table XIX. SufomrWmeoeafkP'rFsOyS in Mallard Serum Sample at 10 pon moor Con So p ee T bUou aT "wae e ppeei SSClomewvsisn Soin hoouwwte oon oieae eTeee me Tw mm amw emi ewe SSSoiommimnisen oooooonn ommemwmp Chins owe oA Tmmeeew ew mmaeew Jam wbhete SSSeoeinmnoomsss ooouwss ommamas Simms owns mn meTeew Toe aaTamwe ew uwee SimoemsotiTJoraamnn Table XX. aSiumfmorarWyeoefPkROS in Mallard Serum Samples at 50 ppm ose CuB e ea bhewe Spaw sutiteegen SS CmtO ioa wWs ooOuoDnm omonnwlseomneww a Taw mbwwne a vtenme 100-001 ppm. for sa boc nd sc en seh fe. Goss Wildlife International, Ltd. we Project Number 454-109 [---- `Table XX]. SorumWmeaerkyo1f0 PFOS in Mallard Serum Samplesat 0 ppm a.i. S>w Coro aSe oeen Apou reasiiegn oSRremmT on E owsn vTmooenwn aTTewem ThTruaamm ootm prS iomntn anpnonirooiwcwonlilhooimoemtn TeS emm y mTimEm ooowrwsd r Semmih inn wwowonn Taomaaewnn em ew TTvauammme eooweee r wr iw een aowmoypn -- Tomo meew mmmaammm ge SSSheeimmaammne oaoowwmme Toannn eTTeneww ammmmem oootwwoe a Se own en mae r-- eSSeatsr wise 0NDm=aNRotpueadnoeaedowtpecckowmasednetciosfd hse hcoenensposdoinngisae050t0rponLie L0Q 2001 uy or ed bodes sds FoC--var 0crey d MQd .D00N0D.gin.shleio loofehe fvevs ininodnid Epes eects cmineg _--_--m--_ Wildlife International, Led. S138- Project Number454-109 Eugen styNo: 023066 `Table XXII aSiu.mfmoraWryeoefP1kF0OS in Mallard Serum Samples at 10 ppm Sor CoDne eeSa S SSumE omowe Tio ooOmTeoNr oooroundd SSpuUsmEimD oooom oom oomoeonmb omomb S"usmmom "simu oOoom oo omonmoo oeowb S S SG mp I s OoOoTmss ooomooedln SSSseeiimomemnleieio ooomvlye omomowsomnmss> simewiiouen. ovens vps pen baumedm aobweo esAmsiee n anmeeedn 7Taaal me iiso a mJaeenn mo maee a0 n3s w oaomn 770m1 mow 20 oinn in memel0l 0aT)we ie ser iTe00eu1 TTeaaebeel)r iiriyo SyaomwoAETyOTaY ms5 L0G = 001 pm. or eb celts andsr [er-- Page 370098 CNL550 s---------------------------------------- Wildlife International, Ltd. "19+ Projet Number 454-109 Bansoc Table XXII.aSiumfomraWryeoefIPkSROS in Mallard Serum Samplesat 0 pp TTSoiyRr a CmBteemS.e Semmes oom own Be hea"n T ACh ow Joa eaabme, et SnSeimsnmaoweiwn amommmme ooowwnnn emmmee ewwaeeeee oss me GSSmiiommmiimnn nowmwma emovooennn Cmmwi ama mn oeelel meelieee ee eae ooimtee ee SaSiimmme Goammmsea Toohaonnn Sma ome mn heeene eeheawee ee ae eoamme SSm lels mi oaommmsaa oowhnnn Some Gm mn meoewm eaweeee me nie a ooem ee - [Ell SE AVERAoG ws:E Oeen NN SD = NeoOpeciN eisso(teckeanstdreiof eheccocotponrsingsteOMciwon me LQ =001 py. or ed bodcls ise SFaoornmldaornsD0c0o0ndmd0).d00o0N00Dysmr.whfs TvoeClloitehee octg cerond [-- Passat coast _-- Wildlife International, Ltd. "vo- Project Number454-109 [S---- "Table XXIV.aSdufmomraWryeofe1Pk5 FinMOallSard Serum Samples at 10 ppm RSrryST C5o dGieemmmmsim GDoiommbae Smhiimmmsam nooomevee SSmeemlmmsii dnoooomwme SSSiimmmsei oohmmoa C Sie ehmmimi onomw tsam Den Phaaen Te T amwe e riasep mowomeoennnsh hwwewee araemnaee la or Ooovweeensss eaawee meTeaummme i Toohvvoeesns>ss mweanme mreaaaeee hhee ooowobens wwaoeem oaeanimee oonn oooboevnn w wr ee aeeat e oh StamunCoRrOr --3 Table XXV. SaudmlmoarrTyEofRPMFOSin Mallard Serum Samples at $0 ppm D e"r4 GB e eSe T Gw"eien phen pttiue SSCesmi aen caoomoev ooommmemananbs weeemm moammm ie Onsen 100-001 oi. for doocnd erv Epes Peepers 6015.72 -_-- Wildlife International, Ltd. ui Project Number 454-109 Appendix XIIB The AnaloyfsFiOsS in Adult and Juvenile Liver and Sera 604503 ee -------------------------- eerie ue Wildlife International, Ltd. Projet Number 454-109 ---- wens 7705: AReread [---- Sw SS E EE E IYnRT vTannTc TIsTES MotCct ohorhysenonaeaie ea sners oneyMyCsEee Bon,he8aiehls To rR areS hy nert.e as eante s dtaae na Tal1. SummaryTah,505 Concorso Ser and Liver m=E | = [meTno]n]wvwremreen || 2T=| |lwoeww|re om) e] rate| To sn| sn | | ww | mow [zxm] e[x n[mmemen|[fa eee] EATe | + even| mem |T | ome |T am JO S-- -- casa --- Wildlife International, Ltd. ue Project Number 454-109 penE01256 JH-- PS: AReSrtywiuteMnalt 0 Astm mrs Tal: SummaryTal,G50 Cancion er andLiverwithCutirs Remord - e 2 d|I | oe |e co mE | memms |v | - TT : Te- , : rs 000 TCnTe 0rae va thyas cot6,505 no or ss Rerenence SussTances ER -- Eea rarnT ase T itOSbene nissmosT|Tccoanre | soon mR ---- coisas 1a Wildlife International, Ltd. Project Number 454-109 Reno 256 PROS ARepdyvrieMoaid + fS I ot maarR , ThsSaate tiye,StobGtEstls sos eae DFETL ReS Tenh e mE r ee A,oor Tarebe:eteWLe AegEeITCSeIoeirRn + a r Ere e Taa AR rmDaya etKaAs TT tsee e rd te o EES Siy e SF EnSfaEpeeene eptatoscl t fosi ee RE Dos Somer oo B Tp F et 1h0e ug 1sEE ArT s snga.e SE inr sSee s Tai 10.50 DateSummary,Fal lardSr AtandOrin) fes o ee amei s snorure | 0n 5 | s1 e| d<s0n| TT eT CN | a Ea mmmmas eS fom emtasose] [eon zssamef re mm [eovizse mere. 01125020678451003240PM remainAdu insmiebe LeLe | 05 | 1 | om | 5 IT JEem o: omne J= e a oom sooa a rem] se asoa | s0 a | + roma|]| forma mmremins 0 T me] Minne Cay fo Suse Wildlife International, Ltd. Project Number 454-109 Report 011256 we 5FOS: AReSpdywic thhiMo ali A BikGE RRE LS 100.3256. 0PPA Fomaie Act 05 1-16ITO 5A 1001.0PP1A 3Fem0al0eAO,A 08 Te EEE fom emo 0 [Em orizsanin Jeso u1068, mPoetw Frm umeiemhiakt |m 0088 ||0a nme] For mrsos sre ses || am mamma ra feor zs as7as Lea1003002,opp Fament | 05 |" 1 | tomo. ose feorizseasrar m amis T lustoniam oro Fermeran | 0s | 1 | em | [JeE2 os ekitoon3mre, oopreou rraenaer saaka | 05 | 051+ | aawmn |ei oe rizsear ms r luss t f1005t252o, a1m07r0orFraeromasoaAeskaaa oToosvs " | iT| s | oxo-- [oeeamsr reTn T o 0 e mm e o a ] m i158avrSoot ari To Er [EomTT ionvez,1Pov oma, gry| G1 es T 0. SoEiEEEREE8 Cor 454113700 0rer10cbr6ny | 01 ee [E+0tT125s62m8s75ea0rra4ss4-1c00.a9a65sm0,o10 PsPM Fumr,eOsgofg DrsaTon 9T %re ene] Eminent abort PE ges ~l6- Wildlife International, Ltd. Project Number 454-109 f-- EROS: AReprStoy cwi ioeMoaln Tabla 1.11505DotaSummary,MlMaladSas(AldOring) er errrs------------ | ram JEOt-1256-28677 orm lasa100:323.. Ads 5s | 5+= os |Tom| [r e204 a shone,oe pp ad 0E 3 |1 1 1 a 50 | 1 5 | JE01-125629552 454-100-302P4o5u, Mie, Adit 0s [Tem feor-aseason lassi06adssopmmmAne | 01 | 1| ceo | CEE T re S CT CN| Em ERETEe RS iIE [Jeeo1z25628r0o0r2 4st5ro4na.m1,01s0o0Pr.oenu3,vM2iok5e.A0sa.ta |0102 0 vo | | e sawmw o| | Tm me a = am eRe [EoEreizseErie lisesowszraoppure pax hn e | o=s| e | i= o |e =oss | EE h Eeitesm rea oa TE Ea m a eeeT -- = EF CE ea -- Cra ney _ , ar Wildlife International, Ltd. Project Number 454-109 ep t01.1256 [---- [Eon125020748 Tesh1000008,0PMMa Otvgang | 02 | 1 | EEeen Seemmmeama a | = PE ---- ---- ------------ IMBvionmenad Laborers PE hen entnn Wildlife International, Ltd. -148- Project Number 454-109 npn rh 705A Represents pn------ 13.180; SummaryFs ladho ptnd spn) EE mypms pesos |CEE EE1132629745 Jopo 238 Lie me EE 11250 1 0 am E------ feor12262o0gmo sivauapa00| e om | | aw | pom al304A ss [T e HT s w [FE Eemampr maaer s D i i e eo ] [E01132625007 pom a132Lo8 ve,a8un tow TT f mm em Tem TFe| EOL132829061 _10ppm ni 0827 Uver,agn| iz | 0| vem | P firso ose m gpm m sero tie s cot = ont T EE mm a T e mea=T|Toe] E01.3253005210ppm a) 3272 F Lhe,sot ___10088 o : --T = 1 Ss Ey EE ae ee -- CALLED Wildlife International, Ltd. 149 Project Number 454-109 ep sovss tn 10. AReSoyp i t Matt tsa renters feouisaboss hopessoot rman |tems |i | ew | 01.196731295_ 0gama)#807Usobgreg| L000 | 1 = B rSaamarre marrooan: Tore a = ] E0o1r132677311250171s0 ov pppmmatt0tc 0 oweolr p|g_1_1000|6 _| 1TmET] re es ro e as r+ e orio st Fnr ong en ma FTE Tea [E0t 138731408 [1p0m a065Lv4, ogg a {evr seraneioTiopomatoorUwotowy| ow [ww] 3MEmirormenidLabora PEE pagers ons --e--e--e------------ee------ ----r-------- Wildlife International, Ltd. Project Number 454-109 -- i os main si ri 1.70 re pestecm 04321 M Lhe, =] Jeo 1322 9620 pn 22MUv acim | 1006 [1 | [E01-132620664 Lopoma.3228uve st | 10336 | Iw pP FPP Eeeems e ssme am eom nssea e e s o e o o s o -- [Fornamets opp 3257 ve,au oe |w| Jeo 1320-25082 a sami sat agingenra foo monb[o0oms5pma1 32s87pinnc s,an | fiii 250aeemasssmrso fatEee f= oi32000 rome sore | ameTo owe | 1r0a0 |0am| Te | rome ry 3MEnvironmnl Laboratory PEE get RL _-- Wildlife International, Ltd. "1514 Project Number454-109 ReponEoL256 nostri PROS: ARepro Sody ithbeMit [EE -- [fEoGr3e1ra13901hloapomta1 9r60e6 ivre,cpg|| voivae| | 1 | || [o m2 z 08M Ure a olpg || v10r87sw || | a a] e | StateueT oF Concuusion BGBFoLs07nT14o0 10mmarTeo(T0o0re)-- o7sdSeotrtoaconceiteftCSnris 4of i oee anheSrEamo hEeoeRt nsSorsasna5cE r nostkneB osTBeoagOne0gp L .hdoTosaedsndm9FaeS irKoosni sThooaewedsnnn arscwaot gbEerneosl Sor mntoTmo ro20 raalocoSraoehrinctonriddnrrgrepetnTseniiadbt0afodeMoacyT h vre oeoo41dE. n,Ss 1S05os1.snBo2a enno ps n ordamnl RnoaARsESkVesSEw SH 4 y so s A LoL a rnmuoyC neiTmfAm hcT,ronSigp20hosuSs e.caoTtrep ese 0cn,or tepC1hy8s 0ao4 h2o0n1end2y30 T CrE an 5woa nTosn e4,noyEa o rCRe eeBSs ow0 oTohe CLEEotmunTtRearn2A mie cptT sgrcoid7S graoutposL ltonoorftese23syT CeFnhAOenorL00adlp3 cB2sk SEoy n aroereeoidehnToo,eTToa e3or15(hEeoS s rok s oyTohmeh Aorn SM irom Laboratory Peat --------_--_--_------ Wil- dlife Internati.onal, Ltd. "152+ Project Number454-109 ---- nn Tal 1 Summary Tbe 17ARe0ged:it Sty otettid rtAtnctn pss 507Concannon Sore ndLie mo Eee | nema] | wAwI| = Tmeam] | mm | | w wwa e || 7 | en me | | wow | sme | | eeww| =we | mee ww | 500| Emmy TI en CORE --- Wildlife International, Ltd. "sie Project Number 454-109 Resor 28 zn 7705: AReSptywicte oald 708 Arnyca eests TRalme1r5 SummaryTab, 180 Concentration SrandLiverwilhOuter Smsmoa Memos Ie E rSAeT ros Yee s SG or oovE era e10sheiF s. A Ot sS teSnEiteEi vcserac r, Strewenror Dama Gunny Dotaorssey ents i R n angeFd onTOo TH1T 172. ile recy. ierind sshe Hania devonofbe pr TT en CriLnas _-- "se Project Number 454-109 Hit ELE ;- Cee he" 3 EoosfsL yoy |i ef2i33i: Insul ce] gg : SiEi9i8 | Bi|g) fzaesgsssnzgesndndsradnasandednel | ax| on | f PE] errno i 458 z| -| scegsrgsecagaray| en FH S8E3R3T [5 |7|gi3o| creornenmrrenneenrnrannneenenen|] ggeen i ER EN B | 15] " (E3f8) i4 .| BF canesgnns asenzs| sc |%8 gGeiadsescccdfEass cdadseneesdd)| 3ix4 | H|B sezszcoaza eraes| sg fli |X EEE ELLEE ETRT FERRE H Ei resenraesisnoans pen 2 5 I$l acavevccngananns| Bfiei|| ag2 61556 "1s ProjectNumber 454-109 An st iFe ge 3 ire 8] 55 2. 238 LI LE 8 & ET LH LN rR 8 8 8 8 = & "io + wae chase? J, Pre Nanters109 8 . 8 5 2 2 } f2e: i y iis i 3 8s fis & HE 8 & 32 8R8882 i . Gnas -157- 34 Pr er454.109 Ep ovge E -fp 103: ; : $334 |8 Sul LEN is ssszssesnsssgsss| ; HREEEE E EE EREL EE I 3g {z e534 i Lf eececcronoccccoe| aa ii i gj TronERTERRREERLa oes 3 Bi ceereenemeeeeeee ann i f 88)| i NHi owBl| 8 {12 | g5f sFsesesss essRsesgsnsnsaysln| uwrsi |i: dnoma ene ivonacn| we|3% 2reeasvenrovneesr saauveaanans|a] gaeens| d3ey1f Mnonnoone nonoan| gun i i vmnveeremcaanaana| f3 ia| B$E8 NAGI EH ti 24 +; fia Re ol vg aE BT ge 2 Che -_ 150. Prt Number 454-109 7 ? He ! an Ng tls asi 5 8s ii 8 #&3 "es i 8 R 8 1580 + wan8 2 A ease _-- -160- Project Number454-109 TERI EY 38 *lae tracer fon gg ee PR [I fgg eed conned a ipakdae"fa] i SHNy fg] ssssgessnngasezn| Hixiild L|H i| | 3fcfrfdddgdgrdsd| sascexssanxasass| 25 | dz il sx 23 AEE i FE eh |%lgy| sesuvacavsavoren| ges i HEE Gf seecssuneasncean| ge I El j2s]l a99m9e0s3 zagse ameiemnsu|sl |2e BEE ii o F2e2a2s5e2 ssamssmszszseoazn)s| sexx| |i3g% lhy| seeRacena snesEa gen iz i2( vezamcsos = sasosa|gas| if |g ore nranan i1 ge |H:y oo rise --_--_-- "161+ Project Number 454-109 Dm i o 3i5t 5 fig gas 87 ,2 18s Eod if i I, OI =, 2 nn II wlgedt ETI 2 3 =p 2 22 i IIIT [III 2 I 3+ um s 8s 2 8 8 2 - Naa + ann nL5eT _-- 162 Proj Numb 454-109 Tm nm TTT o III i rm .s nm 8, [E & 3% QE <2 8 J: 23 nn Lie III : Ga ------r-- I 8 sv 8 8 =z aa + wisi Casi _-- Co. Project Number 454-109 2|if SSHEELLA Ry lslml rg3ieg9 jF e p 3 g if3s : S438 | sy) see sscsvsssesss| az| f 80% o|E| sd ssasdassasgs| o- 2 e1bid Fl [888 sassszzzesss| xc 3 iH; *I5 srsosaaneaansean| ges | | & 29 EH RIPCRRAGIRARALAN] pun i 3 4 55] 8328s sss 83s us| 1.8] fess dus sur ds] il-| saree snares eg) zu i|%s a7 i ge it |i CIARA TBSOIN| AR i $f] cevznenns sagen ans i g i: 3i nme nneannnan 7iis | &ifE 015A -_-- ia Prot Number 450-109 rm I) = . & Tm 5 Fr rr] = SE ig EE 3 nL Er eH 33 QT iy: 3 [2 Wms: 3d I : EI | 2 CE + 7+ OT 8 8 som Azo. nahh _--_-- 16s. Projet Number454-109 rm v 3 I . EI 2 EE +5 5 nm a0, Er - ig Er + is I "i ged I is EI is 82 Er 4 -------- I 8 8 8 8 8 13a wasn aLsA7 ee i" Projct Number 454-109 i(l3E 3k gle 8 | Sag [E| ~|| i $F eu ce ed 411 = SFuri383 | |g) use susasacnasss| EUR L2H] $76 dfdadarsaiar so | : HI i 2 ig Fl TENSE] -| mss ssrsszassssn| sa| I ih #4) sesczsanzaanaran gox | f LE 5 3 & 5 $i sesacneznzuescsan| gaz i SB; E Fl -| S[8i31|| i$1t! : | Ffdi ddd dee es] <a |B] sesse sserass sg em | i53 scsosssmamcoanuanniesaaeeooesn)n| gsansg|| 430 8sRa2n3nesRn%vRn s8sTsRo8nRn|) ggauss ilIi) ET rrr oraaaaaas Bie| 82 BN . Chih _-- -167- Project Number 454-109 z Mi 1 Ee Ti: | Hel IR 558 EH R mm = III = rr] I= Fm #rEe82 [TI 5 5 OT =2 III LI ini i; mm 3. om 5 2 III I um % 8 Erp 8 8 2 6O1559 168 Prject Number 454-109 mm I 3 OI z, ml ht mm se CT] E39 Fr EY iE I 5' 03a 3% (OT 8g=Ai IIE o Hel is 35338 I OI; 23 Ls 28 3 pr =8 El CI = TE T % 8@88 Na + wun = Coan ee 9- Project Number 454-109 Hi L 2 . E] 10d veal ggial (poy S Sigripd| 2|kg) g3r3s% sfndisaeurRgisigIiIrGeYsl "i1f 2EES i | sas sasazggssssg| & i i 3} #48 grzensaasataAnIAN| gas : : i& aPE7! iBoEl sesenssnauszesas| gas 2f Ja) dEEEE REIERR ad) | HE [ Hig c$5a8n5a8eeno%s55s8e8%ne8s8| ga3a i|l) ii gREsecRsA 55AAR| FAS iiii 2 Z3il nemevereagnnazan) = if] i3e s38|| 5s& % GoAsht [I -------- BUR Project Number 454-109 = = |= ie CE = GE I = |: tia #lgi3t 28 I =25 `3 Ed ps 5E3 I sf3 mm:LI ti CE = 3 + min DT gs I 8 8 W+awaosn & = GOAT _-- me Project Number454-109 TE I 3 mR z (ID 5 i I= 8g OI + 52 i mmmm-- BE TE "iq gid I 5 Is CT 54 I is 53 mmm----------n"n I= 1 I TFs Wao / 8 am rR = Goth --- . Projct Number 454-109 Ila s3sd ||i ~ Epr0a0c |ul | omo ann ) 3i LES ETH ETSEET : tii il - resessasiazanzaz i iE3yii 13sB]s 22se3s 0HIAEAIFISIN| B2ge2n] |: Hogi man) |b 2 $i eemcasmnsasacsan gas HE ! 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FtIe : ; Fo]fcmvauno=nn -E .|2 3|& i=) Go1551 _----m . Projct Number 454-109 iil s2eseverss |; HH HA. : 3ogx i3 cerererene]g ; 3H3EM HEi} cowreC ncneo sia sd oo. Pil$8} "3% 3 g| 000000 0 creme] i & | lg) emevmnevean ged g| rescen| c|e 82 5z Go feavannonnn i: Pt g JE! "i2d8 CNH ---------------------------------------------- ne [---- i 3823338 2e |g i Eben Ios8 83 Servwmnrnenly $483 [LF 2% 435a |Ze i ew or erm oe ow oe a : 1743 gl creme HE al eres] Pig al creer afd FI i) EPoOsELR EY III seenmounen]afd 2 III +3 |3]fcovanaomn ig : it - 1 i 8 i 2 CRLGRE _-- i SAI REBRSAL [ g Ele ogg | 5fF rd [FC Paul oF ox NE Bag | 28.8 gf To Bf Pr 3 eee corer ere |g rerels] cocococccealal E oooiiiiinl,e lis cee ane ee wow sf52 fil i it i lif < BG E-tord _--mm rier sea 3 fers ases |g e | coool PoE I ogi3 : H 2 co emco enrcn oang a|y SEE OR . MEE: LLL] EBC EEE | fs oreo & : H cece eee "le i 118 fal femeemeen ii : g i i 5 001555 - wre rotNant 454109 3i] ~scerszeaz|sg sab i 3s EE $x2i3r1y i Piyf LEz REI ER a[f.f Cee | :ff arecerores asf ls3 FR2:EE | 3 ea eld IIEiy cbeeomveeer n Sg a l3d Fihe 7i i2i CNAHRY - rN 318 -188- i en): a seomer ren' pal| SR PEEL LiAE LL(), Fg3dERsEd] a|glo ema a |s 2B8A FE ap 8 al c eme mer aerny aa|aalfg eee od FY EERIE isl . 7 |fecavomno=nn i # afd 3l!g8 3 COLE I awe ProjectNumber454-109 exgaaReIEa g 3.33 ial g.88 |g 842 Ze Mat FBRaEnSdE |] p 33 B EE: : 588 |] {5 . & gl g cecJ a]R or = Cee | oes cee e] Ts{ ccaeecnmeeeencocnencla|]o] ene algd ) Tlis ccc" ge commons |g HE Pi ; is ls wmerevur ely 5 f : lis 2058 flee EES Be ; sasageI ess | V g EEff Belg li E5l4y% iq # Coil cf menm omnes|, 13 (fee| iy: frocnnnanns i : Fi : 3 GOALS! L191- Project Number 454-109 i snaseacza |g tii | Ce) : BE ROSEESNNN I ied Tote . 1041 JE ei.f1fd FFE tF sh1ia5nl8 i Eos dg sell ] |g) eoo woom ak |3], oT oororo n "lal: io:f E sf p ece oe s iN;3 ial eana. dlEE v &g z i i 60LEVE eeee ----E---------------------------------------------- "2 J-- 3 23a 8RTRIT |S 5g al 3 ig gl PLeEB8gh [iolffe] Bf 3ixR2g8 "la8 Babs |] cee enna]. cen |s Ccn0vioieivielnlnla0l0sx Sesvasanns Ig 2 ceaeeanenns] ity g| ceeceeeeen|o]d l 3 ene : pi : : .3 Fo fcavannonnn F|F ir 2 : Gnas? -- 193+ `Project Number 454-109 i 24 = SAs Aa R : HE Lif if i E i ol BE 8 RE BY | ||rrreeiee 4 siren 51 8333848 HERARS # COLTS ee J wf oor Co 2g 3 si hia r lir sai g1l3 FRiY:| |-C|3arieiiaii : : Bs 666 66 8 1 : i} i El BARRAABRARNARAAR 7s GrAn7G - se jcNar 454109 $i] sannc aes 2s 2x] og i X282% 8$T 8x3 23 ir H <| sasasssss ssesaam g S| BE SE:E ERF AEF i$l9 "Sl 5R-ERTEFE BEECHES _|2| sss=sssss =msrsss i2 3 is HE TEBE TCEUE REECE 3 iF [Hs smpener meee 85g 3 RXRIZ $92 257 AR i PE i HEEEE E988 HE 3] EE aun vey AEE i : ipsis Bj ceizesass ssesse :fz iey "| RTEETUERE EEA: sf i 3 | RETFETETY ERE RR i:5 5 8 @| sasssssss ssesss (35D #| sss=assss ssFEsEEs st S| BTTFC EETTEETEET 23: i [bd hdd Bi: 382 106+ Project Number 454-109 if s32 ngsasass-Isa| g S| EEE EEEEE EEREET i ZFl| DEEETEETZEIESYEREIRIERRAENEGEEEE 8g |F|(z=|] zSEzTzerUsReUrRsEzERcRaAzReEsYs sud 130 $85 |F|F| SEESEEATERSEEAEE Esq |F| FRITIRERARRY AER veBs lf 2] cocsssrasasaases ES8E5E8S |i|5| ssstizaaaaszaas F15:8P03 |I s| sFEoE RsssR rCCaEnEsEI sEsEaGsE eRsEsS BE ] 3z ig is Be i1i34 [jHiiHn: i. | EECEEEEEEEICEEY 52 || meee (HE Bol || esoermmsznenees 80 Cl E5R EBSzsaAA 8 HH = TE FUTURE iil$52 HEEE EEEE REE EEEE RE] 58 COLTS _ " Projct Number 45-109 Ff] "of i| :i Z| I iT HE H ;. SE 1d: fi I; : il, cade T1 a& JF | if 5: | =|| 3: gji =z| : i 2>| | g] wauac ax zazes is re oan geese weBsF BgiEtR &al%[}% 5E FR5E5E3R55E 3T8EE5R BE EECHEI R E FiRii Rie FEE EEEiE SECEDE A FESE MBAEE RA LA BREE ene ssBsoRrEeEsrsngEgs E yeeeE sss | ! EEE EIE FER iE i scErszsazseases sereenn if 531 EEE REECE ! M ssS ssssass Ta IEEn EE 3i BETETERE ERIE IH 5A55L 335L 555A33H os i i zg sazssazz zeEmaTn: i i tezasenanaainznaanzys 9is 010g ------------------ 10s. Project Number 454-109 i ays mgsnnssxevaa| 3 B| #88 2]a2AxsaNENR|6R = 1 ame . x pi ZF SEEuganresaaras 88 de] NILE EERE WEE svvsrenrnaranven es JE;- 1| mre a 1 Ea a | i nds tla) sri srrersracnse z ii: I i ad 2] FocETERRERRR EERE if 35 Fl ERETEIREEEEEERE H id F| sgEsmzEssasEzees i CIEE i #ETTTTTRETTTTTUC 3 i Glringg --_--_-- 199. Project Number 454-109 OffsprAipnpgeLnidviexr XVIII Weighs (s) from a Mallard Reproduction Study with PFOS Control (0 ppm ai) _---- Male _-- Bird Liver Female Liver 1 2 3 s s 6 7 Mean sD _-- 19845 ois 12.7400 _ 14534 4619 18179 1219 1.862 1513 15982 48 13.485 Gow 14951 Mean 14.826 2326 sD 2917 _ --_-- ---_-- Bird 10 ppm ai Mate Female Liver Liver 1 1418 13077 2 10819 3 1719 1136.049059 s 181 5 6 11.558 ns 7 --_-- 15334 ------------ Gm Mean 13652 sD 4398 14.068 Mean 13.944 2183 so 271 _-- --_-- "The above diffrences fom the conrl group were no statistically significant (Stdents Ces), rror------------------------------------------ Wildlife International, Ltd. "200- Project Number 454-109 Appendix XIX Changes To Study Protocol - he fol"lhoiwsinsgtuadmyenwdamsecnotnsduacntdeddeivniaatcicoonrsdance with th study protocol signed on January 22,2001 and 1. TcCWohholelelenlcptriioaotnvnoicloooaflfbsllwbeel,aocsotfedeacdmasleatnimssdpaselmudepessletfdosourarwdiaenndagrlfytosshuesersotpaas0ni.drbshoiAfscdoobdpliailretdochsntoetlldooygef,caoactlmhheethxepaesrtmoiltgnoorawcotoeuilrpondwtiaorgseobsemtsimmvaeealniadnatcedadtiitn1oe0bfdir3edf0sos,8r birds. 2. Tcroehndecuecnpetrdroatbtoaicoosnled1w0aups2o0n.apmeepnamdme.ednattorltrhteeedbudecgmitonhrieanlgdiiteoytfaraWynedeckoSni3cgneosnftothfieoontesftwo,r tThhaee 1ct5hoe0ncep1n5pt0matpai.ponm wtaea1sst Concenraton 3. TirheheceonvpedrroetodofcofWllewelaksf4oaolmlfoewntihdnegderset.douccWuttieohcnaknoz4febtthohedeyi2tW0aEprpIymghacoa.nncdernoetbrsasttcmimeoannttifgoornomdupa1a5s0Isn1od0oi.nc20taespdppbrmiarcdatsi.cpaldBaifrntdexsr ere cuhnized without collectionofsamples 4. ThSihegenespnrodoftootfcooxWilceiwetayksa6tamtoehfentdh5ee0dtpetspot.mcuT.ah.neilzleveeevle.tlhewBa0lsooptdpe,rmTmeaiirn.,erdielsdetumeaenn(dt0 geforacoatulepanams'pseiocoen,dawshpmernaoctraivcaaayliabfaltneedr wre calcd from 10 pis when he ve wes terminated 5. mhTehimebcrpkraonsteo.cmoealalsbwuuarmesemneanmatensnd.dyeodlEkg1gn0sdadsiedlrecceoteldfctufoosrneLfoootfssaAan,amlpBylseissa.nfdEog1mgswaeslrceeegcsgecspdrsfaetoleemdctLendoosfoCrrshocealug.ghshheell| Wille apencd and the yolk was sparted and sored rose for posse anys 6. Taivshaseiuaepbsrloet,aoncfodlomwf1ae0s.oafmfwesnpardriendgcotcolalcaehdctdreodcolfltoerchticpoononsotifoblllegordon,uapsyaesnled.ct1e|d0ptTimsusue.ess.aenwdaetrfeeecanlshswoao.mopplk.eesn,B.lwohoeodnF, Histopathcogical examination 7. TplrhoietopacroolltocmoTlehhewoadsusaewmeoerfndamenaddeit.notremeamlovseatnhedrdefewraesncedeseumbestdanucmenmeumcbeesry(f52e6) eofmncmpeangtss2 1o6f hthee 5. PCTohprepaoprr.uoittoonec.otlcowTniacssesnutaermsaetnciodolnelsdecwtioleldiofnotdribhceitssthsoiappmapptelhdeoslocgEoiPlclLsetfxeoardmeifnxoaratmainioannlayfstioismobwnubo.tiudlwsdilibnbeetshihesip5pp0seedadn(d0of31.5M0 G1 ---------------------------------------- Wildlife International, Ltd. "01 Project Number 454-109 9. TEnhveirpornotmoecnotlalwaLasboarmsetnodryedfotrosandldysAstsofchsmaemnptleA, he analytical phase plan to be used by 3M 10. TEhnveirporontmoecnotlalwaLsabaomreatnodreydfotro aunpadlaytseisAofascamhpelens. A, the analytical phase plant be usd by 3M 1. pTehaekparroetsocporlecwiassioanmreenqudierdemteoncthaanngdethAetmsecthhmoednfoAr,atlhec3lMataniatlhnyeticcaalllrpahtaisoenpcluarnv,e.o redefine he 12. TrKahdceekniipnrcgoatntoucomonlbeNrowsramsabsesraocm(ieLantIdeMdeSd)wi{ttoohrttihhneedisatcnuaadltyytitachraeel2pm7haa2is5ne,p3lEMaO.n1-rE1n5ev4is3rosannmdeEn0Et10a-1l1-2130L65a5.b.orAadAtdorndyiooPnrsnoje.ct 3M Principal Analytical Investigator vas chnged fom Harold Johnson 0 Lis A. Sivenson 15. TSuhseanprAo.toBcoelacwahs mended to change the Sponsor Reprsentaive from Rochelle R. Robidea to 14. NTheewtpreodtocol was amended t change th Sponsor RepresentativefomSusan A. Bach to John L. 15. ccFlxelacteehtpeeirdo.snaomfTpvlheesprrwoestroaecmopclloliel,decstneuod,eewqshuacemnoplallveaeicclotalbileeo,cntreodohmfosradeaultnsabmiypdlesws.feorAmndowithiiwocenhglihyss,duewprisiahomrtp1hleebsewienrge Placed in sample conamers. 16. cOonmiJoullygr1o2u,p,20w0a1s, itnheadtveemrptenrattyuoretirnecborrdedo evel 523, housing offspring from Lot of the 17. D2i0e'taanrdy sJaanmupalreys c25o,lle2c0t01e.d prior to being procesed afnoIrDnaiamayld0y,sossfaWmoepnlcekJsan1wuoeafrreyhes25os,rte2d0w0ei1n.rehneoTthaensatlopyrrteoidtcoafclroolczheinenmbdesitrwcyeaeJnbtoJhraasntudoairreyy Samples would be sored frozen unl hy wee preparedfo analysis 18. wThrreeinadaudlvterbtiedntslyinctuhteha5n0izpedmst.f. htebset gcionncienngtroaftiWoen,ekth 3 poma es eve bofn1ihn setnd2u4r3inagndevhbearpaasiirainopeFn 214560, ~eal3AG -_ m2. Wildlife International, Ltd. Project Number 454-109 Appin XX Personne InvavedinStudy manageTmheentfooflltohwsinsg ykey Wilde Intemational, Lu. personnel were involved in the. conduct or A(v1i)aMnTaoxicDoalbosg,yWide Toxicoloist @ JoanB. Beavers, Director ) LindRa. Michel, Manager @) DismaTempleLaboratory Supervisor (5)) SLeaanmPT.y.GFarlelya,ghSeern,ioSreniioorgBiioslogist (7) someGriSnmireBioslog,ist A(na)lytWiciallClheaBm.iNsitxroyn, Ph.D,Dietor @ RaymonLd. Vantoven, PD. Scientist Go1551