Document rBmGyD30nXQ140dkNY1wX7N9G
pros AREPRODUCTION STUDYWITH THE
MALLARD FINAL REPORT WILDLIFE INTERNATIONAL, LTD, PROTECT NUMBER; 454-109 SMLAB REQUEST NO, 12723 FIERAGuideline 714 ECDGuideline 206
AUTHORS: SJWVeoakaomPnno.b.nGedaDrl1le.avgVehaenfrioven STUDY INITIATION DATE: Janay 2,201 STUDY COMPLETION DATE: Augist 21,2003
[-- Env5sivrCoothnrAiavabenoniry St Pa Mines 5106
Wildlife International, Ltd.
E5a5s9o8n,CMoamymearnd D21i6v01 io) sito Page or202
1005
Wildlife International, Ltd.
Project Number454-109
2 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT
SPONSOR: 3M Corporation
TITLE: | PROS: A ReproductionStudywiththe Mallard WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER: 454-109 STUDY COMPLETION: August 21, 2003
by the U"Sh.e EsntvuidryownamesntcaolndPurcotteedctiinoncoAmgpelnicsyn,ce40wiCtFhRGoPoardt L1a6b0o,ra1t7orAyugPursstct1i9e8S%taOnEdCarDd,s Pasripiupblleissheodf G38o5o0d, ALgarbiocrualttuorraylPPrraocdtuicctei(onENBuVrIcMaC,/1C0HAEuMgas(9t5)198147,)waintdhtshepafnollMoAwFinFg,&x$ce9ptNioohnsS:an Notifcain No.
arenpdor3tsMwCiCololmrppbloeersattueisbominigtnaeerddansonetpdariadntacetlleuydd.erdep1artthsefrorepbrl.oodaSnudmmtaisrsyueabainaelsysaerse cinocnlduudcetdedasbaypEpexnydgiecnesRensdcarfclhl cRoensdcuarcctThehdanusanltdyutedirycawolanpsorptcriooontndosuccowlteerd(eWucniodneddruecmtueIlntdtiepamlnaedtepirroontwaolco,olsLa.did.iTthsotenuadliynsdenipueamrabanteedrp3r4Mo5t4oa-cn1oa0lly9st,i(cBnaelgypgoetrnhieRonEesxsywgceerhne study umber 023.006 nd 023.070 cpoondructteAsdnaeilnypsaaecsc.oorfdaengcge cwointehntGsoocdonLdaubcotreadtournydePrraEcxtiycgeenStRanedsaardrsc,hasntdudryeusumsbeorf t0h2es3e07a0nalwyeerees naroe
`The analytical phase performed by Exygen Research under study number 023-066 was performed in compliance with EPA TSCA GLP (40 CFR Part 792).
STUDYDIRECTOR:
SAPD.ie
Gallagher
20,
Senior Biologist Avian Toxicology
3s 0a
DATE
SPONSOR'S REPRESENTATIVE:
wtNewsied
DA 2
C1006
Wildlife International, Ltd.
Project Number 454-109
i QUALITY ASSURANCE STATEMENT
bGyotohdeLUa"SbT.hoirsaEtsnotvruiydryoPnwrmacesintceaexla(mPEironNteVedc/toMiroCnIcAComgHpcElyMi,n(cOe460)wiC1t7Fh)R:GaopnoadtdJLa1p6a0ab,nor1Mt7oArAFyug,Pursa3tc9ti1Nc9e8o9Sh;taaOn,FdaCrNdDostPatrsciaspcuriboplnliessNhoeo.df Sda8t5e0s,aAmgyrifciunldiunrgaslwPerordurcetpioornteBdu1rahue,S1t0udAyugDuirsetct1o9r84a.dTLhaebodrattsoroyfMaalnaageumsenatnwdesreecostfiloonwasnd the
Acnviy T-- ot bumee Mati forinion pirBodAynweight ant Hing body weight oLny ldbody [ mn-- Candin ASeReDrS ds cory dAnaynieda nd
acnongo a nd Fi sgrt
DATECONDUCTED Jnary 22,200 January 30, 2001
Febuary, 2000 May 24,201 Mayat200 ays, 000 ye, zm01 August 14815, 2000 SOcotoober2, November 28.5, December $7,102,200 August 20,208
STUDYDDAITREECRTEOPRORTMEADNTAO,GEMENT
Jy 00 aL
Jaa 23,2000 ay 31,2000
Fema tn May20,200 Mey 1.200 iy. 200 ye, 2001 AIS IO
Feary, 2000 Mey25,201 Meya1,200 Jaye. 2000 Jayas, 2000 Aus, 200
Novnber 5.2001 December 12,201 og
November, 2001 rch 1,202 Augz1, 2003
QRoubeyr AssMurcanGes Representative
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Cndooy
e------------------teteieer?
Wildlife International, Ltd.
REPORT APPROVAL
Prost Number454-109
SPONSOR: 3M Corporation TITLE: PFOS: AReproStduduywicththeiMaollanrd WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER: 454109 3M LABREQUEST NO. 12723
STUDY DIRECTOR:
Mo LL SSeenaino.r BGioallolgiashte,rA-vian Toxicology
:
&
4.)oveJ n >
Scents, Charisty
MANAGEMENT:
_ Director, EAv- iaRn Toxic-- ology ,'e
ldV b. Nir,8
DirectorofChemisiy
3! | 0x
BATE
BAsThEhalps
sree
BatoH
611008
Wildlife International, Ltd.
se TAOFBCONLTENETS
Project Number 454-109
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Wildlife International, Ltd.
ProjctNumber454-109
TAOPFBAGCEOSLNTEENTS EOOifpfpiSihnnegllBLToiHvdEeyRrBWW EeSiSght eCh II r 227] COOo TpNLOtn BVREiTaVe 0BSnS ONSg.... on12 III7 37
TABANDLFIEGURSES
Table 1. MferoamnaMMeaasluareddRCepornocduecntS(itpoprynmaa.ti.)iofoPnFOsS in AvianDiet
30
Table 2. SARuemprmuoadrhunyceoiTfoegnGsrhSotssdyPwatihholoFgFiOcS!,OAbsdeurvaBtiionssFfoooumnldDMesaldErudthancedrn
Table 3. MeRaen ApdurloBoSddtyuudWyewciigththtP9iROoSonma Mesrlred.
on
Figure 1. MReaenpAdrultoMidSsituuBcyywtiWhiePioRgOhntS(5) from arMaell.
SU
Figure 2.MReaenpAdumltoFedmadluyesBwiotidhyFoWFOnSe9ir) orgmaMhalltard _--
a
Table 4. MReaenpFeredoCodnSswuuamcypiHtoDniP(oRbOniSde) romaMillard
35
Figure 3. MReaenFperd oCondsudumypwctiitotnhiF(FgoOiSdnday) frMoallmard So
----
Table. RSepTureomduTcmetomiafioGnarrtSoyotsnyPawtiholPogRiOeS,a ObAsdeurvaBtiiorndsFuobmniavMealdlsed
31
Table 6. SMetaunywLiitvehrPWROSe(9i)fogm ahMai llsard Reproduction I
38
Table 7. SRepruodumctoifmoRneapdroyrdwuiycthePPHeOrSformance fnaoeiamlrd
-
n1010
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Wildlife International, Ltd.
ProjectNumber454109
ae TAOPFBAGCEOSNLTENETS
Figure 4. MReaen pReprrodouucdtiyvueWPceifltorPmiFaOnoScenfom Mallard
AO
Pecetags (4),fom a aallrdReproduc-- tion S--ywith PFOS........41
Figure 5. MfreoamnaReMplrlodaudctRiveePpeforrmoanSdceUu,NcowmtilhizPeoOdSnas.Percentages 4).rec
Table 5. MeRanEEgPg SIheSlOtlTuhdGiycWkUiocksEPsReoOsSunrments (a) fom a Malls ard rk
Table 10. MroemanaBMoadllyaWrdeiRgehptro8d)uoctfiotneSludiywniagtnhsdP1ROS4O-ldDSuravipvioyrs
th
APPENDICES
Appendix 1. Conificateof Ansys
oe
-- 5s
Appendix 1. Diet ndSupplcat Formulions
."
ADDED I. Die PODAMO
$0
Appendix IV. The Anaof PlEOSyinAsvenis Dit.
nS)
Appendix V. DigofTrestaLaymo cc.
--
rn?
Appendix V1. Reproductive Parameters andReps entiation
tt
Appendix VIL ARdueltpBordyoWdSetiuugdyhcw2tihiGroomanMallardPROS...
TS
Appendix VIL FRoeopdroCdouncsiuomnptSitoundy(wbiitrhdPdFsOyS)..r. aoMalmlrd oe
tl
Appendix IX. NiInldaivnidduaRl GerposrPtohlSdoguyiscwlitObhisPeRovOtinSons fom . 5
Appendix X. RAedpurlotdLuicvteiroWneSigthdtsw5i)hfoPmRMOaS,llard
---- wo
ADDED XI. HiSODABOORYREPEL...
89
[DETEY
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Wildlife International, Ltd.
Projct Number454-109
1 TAOPFB AGCEOSLNTEE NTS
Append XIA. AppendixXIIB. Appendix XII.
Appendix XIV.
Appendix XV.
Appendix XVI
Appendix XVIL
Appendix XVIL
Appendix XIX. Append XX.
The Analysisof PROS in AdultRedBlood Cll and TheAnalysisofPFOSinAdult and JuvenileLiver and RReperopducrtiovedSPeturdfcorwmtianhiceobynPenPfRaoOMaSlml.ard.. ReMparlolduudcRtivee PrerofordmSanutcdecbywytiWtehiPeEkoOaSnd Pen fom EfrgogmShMelalTlhridcknReepsrsodMuecatsiuornemSetnutdsyw(imtmh)PpReOrSP.enby Weck rMeoamnaHMalalutd RBcoedhypWeieigonhtSdgt(5u)udpycewriPtthiPbeRoyOWSnenck MWeeaenkf1r4o-mDayMaOllldaSrudrRvievorpBordyoWedSigtuhtycw8i)ttpeir Ponnby ORffeprpinrLioveSdrtuWudeyicgwhitttshiP5)RoOfrSno.m.a.Mallard ChatoSntudgyProetocsol. ne Personnel InvolvedinSudy..... _
Ss... 128 Sr. 41
158
en 76
193
195
PROS... 197
199 200 202
crioaz
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Wildlife International, Ltd.
> SUMMARY
Projct Number454-109
STUDY: PROS: A Reproduction Study with the Mallard SPONSOR: 3M Corporation WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER: 454-105 TEST DATES: StExupdeyriImennitsailoSnta-J-anJuaarmyar22y,22012001
AFPhdiouttlotEsTgigmeusrlSamteiitonn-Aa.ptAtiupon1i3dn,243,00.3020110,1 2001 EBAinxoaplleoysgtiiicmcaelataTTleerrTmmiiennaamttiimooinn -n- uAa=pyAtepo26rn1,12,102100,1022002 TESTANIMALS: Ma(tralsplaa tryncd hos) AGE TEST ANIMALS: 24 wesks of agthee iniaionafthe est SOURCE TEST ANIMALS: W1h1i3sWtlaisnhgiWnigntgosn,Snec.t
UHsnkovIeL 61041-3509 NOMINALTEST CONCENTRATIONS: 0, 10,50, and 150 ppm as.
RESULTS
weTfhrfeeercetsawsue,proencwaacdamutletmebnotderlyealtwaeetdiegdehftmfeoactrtsathluepeoS,On aofvneederdt1c5s0oingpsnpumomfpait.oonx10iasttcctoahnencde1tt5rr0aetaiptopmnemsn.t4e.1sT.thterode oFcfoenpcrteoondxruitcciiov,ne.oprhThateeos$fm0ehanentdrtels1ta5.t0eTpdhprfmea.octr1estumpcooonn.rceenabtiormdaeytnitowfneseitwgeehrd,emtfeoeodnmiincaetocsndmvppeirurtongteohre aienpycrroeRadasuevcdeibivenecpnkedrrnfecoeermdoafn1scmearlaalimteheentt1s.0 opBrpamsmeadl.oenttishehcioensct1e0nstpoGipoen.ar1.dHtoetw:ehvceeorn,1c0ehpnresimownaesttorn diCuhorenicndeigcnttahtiswiaossn,yd:ethteernmoi-noebsteorbeede6le5ct(hcnonc10enprpami.ona..o.rthmeallloawrdess ecxopnocseentdraftoioPnFOtSed
n1023
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Wildlife International, Ltd.
Project Number 454-109
oe INTRODUCTION his stady was conducted by Wilde Ttemational, Ld. for 3M Corporationat the WidIfe ner cnaotnidouncalt,edLird.oamviJaannutaorxyic2o3l,og2y00f1acinliitly iJnulEyas2t6o,n,20M0a1r.ylaRnadv21d6a0t1s.genTehreatbeidlaotgWiiallpdoertiIotneorfatthieontses,t Lwds and acopyofthe ina epot are led under Project Number 454-109 in archives located on the Wilde Intemational, Lu. se. Biological speciarmeestnorsed at 3M Corporation, St. Paul, MN 55133
OBJECTIVES ofdieTahy eaxbpeocsutrievootfhteipsotsatsusdiyuwmslaeovfaPlseurafltuotrhoeofeetacntesSuuplfounrtihc Aacukt(hmearlelaafrlder(rdensfperltoea3dyPiFnOS)) over periodofspproximately 21 weeks. Effects on adult heli, body weight, and feed consumption were evaluated. In addon, the effect of adult exposure to PROS on the number of egg ad, etl, eHmisbtroypoathovilaobgicliatly,exahmaicnhataibointyo,fseloefcftperditnigssuessuravnidvala,naandloyfesbglgeoosdshealnld itshsiuceknseasmsplewserweerecavaslounuesde.d 10 evaluate the eects upon adults expasd 0 PROS and hen offspring
EXPERIMENTAL DESIGN
rcatmeMnatlglraorudps(.80Thmaelteesstacnodnc8e0ntrfaetmiaolnessw)ewreereselreacntdeodmilnycodnissturlitbauttieodn iwnitohotnhee Scpoonntsoolr,gbraosuepdaunpdonthrtehee
reuls ofa pilo eprodueion sky (WAM toxicity information providedb the Spenser.
Intrnational
Li.
Project
Numb
454-109)
and
adions!
_--PRO-- S Treatment Grou-- ps --
Group gpmai Nominal Concentration PGernosupper EHxetmraatPoelnosgy- M`aBliersdspeFrePmealnes
T 2
(Conra) 0 10
Is
3
11
16
4
:1
e --_43---- 15500 1s 1s s4 1| !1
_--------
Wildlife International, Ltd.
Project Number454-109
n-
pen. FEoaucrhadtdsiatimoennaltpaindscoofnbrilrdsgrwoeurpecaodndteaidn1ed c1a6chpatiesst oefvbeir(dfsorwithfooanleomfa2l0e paanidrsopneeflevmael)e apnedr
maintained containing
for collection of blood samples iter 10, 50, or 150 ppm ai. of
during PROS.
the The
test, Three treatment control group was ed
groups were fed diets diet comparable 10 the
rcatment groups, but without the sitionofthe et substance.
Because overt signsofoxcty andtestmentrelated mortalities were noted at the 150 ppm a. test concentration, the dietary concenrtion forthe 150 ppm a. treatment level ws reduced fo 20 pp a, of PROS a the beginning of Week 3of th test. The 20 ppm. level was terminated at the begining of Wtheee1k50Sof7t20 phpmesait. dtueesttocotnhceenltirmaittioenrweiclolvbereyroeffbeirrdstoa3hitshefs1t50copncpemntraa.ticoonn.cenFtorratrieopno.rtiAndguplutrpionstehse, 50pm ai. est concentrationswere euthanized at th endofWeek 7 ofthe test dueto came ested `mortality and over signoftoxicity. Therefor, the tet was completed with te control group and the 10 pomai. test concentration only.
AI adult birds were observed daly hroughou he ts or signs of txiiy or abnormal behavior Adult body weighswere measuaretedtnitation, on Wesks 2, 4,86,a,nd ot adult termination snd feed consumpion was measured weekly throughout th test At the beginingof Week 11, the photoperiod winacsubianicorne.aseWde1e0kilnydugcesegwgeprreodsueclteicotne.d bFyoilnlcioswriimnhiegntseadtroafwefgr, pergoduschteilont,hiecgagcsswsemresssturwcemekeltyafnodr lf remaining eggs were candied prior to incubation to detec cg shell cracks or abnormal eggs. Eggs were also candied twice during incubation to dee infer eggs or embryo moray. On Day 24 of incubation, the eggs were placed n a hather and allowed to hatch. Once hatching was completed, Hatchling were removed from the htcher and the group body weight of he hatchlings by pen was determined. At 14 daysofage, the average body weight by parental pen of all surviving offspring was determined. Upon completionof te te, sttisical analyses were performed to determine statistically significant irene between grovs.
621035
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Wildlife International, Ltd.
Project Number 454-109
we
MATERIALS AND METHODS ReprodTuchteiosntuSdtyudwyawsitchontdheucMteadlalc.ontTihneg ptroottohceolprwoacsedbuarseesd oonutlpirnoecdediunretsheouptlrionecdsi, t"hPeFEOn:ironAmE.eHnatazlarPrdotEevcailounatAigoenn:cyW'isldRiefgeiasntdatAioqnuaGtuiicdOlrignaensisPmesst,icSiudbesAescsieosns7m1e-m4;GOuiEdeClDineGsu,idFeIlFinReA2S0u6bdisvnidsitohen ASTM StandardPratce or CondictingReprodhctive Ses with Avian Species" (1.2.3),
Test Substance
signeTdheWitedstesubIsnttaenmcae,ioPnRl,OSL,d.wadsenriefciisvteidonfrnoummb3eMr 4C6o7r5pourpaotniornecoeinpt.OctTohbeerte2st9,su1b9s9t8ancaendwawsasa
Whitepowder nd was identified 3: and expiration date of August 31,
FC-95; Lot 217. 2006 (Appendin
The test 1). The
material hd a est substance
reported purtyof 86.9% was held under ambient
ctioonndsitoifonsthien fleotckesdubssttoarnacgee aitntthheeWdiictfweerteeandtjuisotend,toLu1.0i0%iacstiveiniEnagseodine,n.MaylTahenrde.forCeo,ndcietanrty
concentraratexiproesnsesd a paperrmitlasn ative ingredient (ppm a. in the dic
Test Organisms
WashinAgpipornoxSitmealt,elyHa8no4vpear,rsIoLf p61e04n1-c05e09m,alUlSarAd.werAe tpuhrchsatsaretd offroamceWlhmitsitoinn,gtWhiengmsa,llnrcd.we1r1e3
ahaptcph,aprparohaechaibnyg atnhderphfiesntotbyrpeiccailnlgysienadsiosninagnudishhaadblte fbroem wiulsdtdypne.anyThperevbiinodussweestriengf.roAmLtthehssarmte
opefnacacslsiimgantmieonn,t aforraenacdhombinndu.mbmemregdeineerlatyinpgrofusncttoitoenstinnaatisporne,adaslhlepet oprotgraamswasbiussedwetroereaxnadmoimniezde
1f0orapchlysiimcaaltitnojulrabeosraantodrygecnoenrdailtihoeanlst,h,oriwdese hcttsddied tohte swepiaghst hraenglfo eithheertdesute, tworereyxcolrudaedbflroym
he study. in weight
A birdswer 24 wecks ofage a from 781 10 1444 grams at test
st nition (ist day inition. Sx ofth
of exposure est ie) and ange birds was detmined by a visual
examinationofthe plumage
entation
and
Adu ids wee denied by individual leg bands, groupsof pens wee denied by project umber and
cachpen was concentration.
denied witha unicue umber, Al eggs id din the study
C1016
ee
Wildlife International, Ltd.
ProjectNumber454-109
Ry
`wermaskedwith the pen umber sin permanent nk marking penfo identification. identifiedby wing bansodthst they cobeuradced 0 theiparental penoforigin.
Hatchlins were
Avian Feed and Water A sue bids and thei offspring were given feed and water ad bin during acclimation and
tsepsetciinfgi.catTihones bbaysaAgdwicatyfeIndc.to(AbpopenaddiuxlsI.andThoeffbparsianlgrawtaisonfcoornmtualianteedd ato lWeasit d27e%optreortemiantoannad,2.Li5d%. at,andnomore than 5% fier
timesto"nTeheusbadsanltdihetfocromnutlaaitnieodnoafptphreoxibamsaatleldiyet1b.y1%Agwcaalyc.iumW,idleeritvheids efvreolmoffceeadlsctiufufmsiansudfftihceien0t.f9o%r
rowth and maintenance rations, additional calcium i required in the ationof breeding birds for egg
sthheellbafsoarlmatdiione. foTrhetrheeforaedu,lats addTihtiosnanlis5e%d (tvh 1)caflciluimmeslteovneel (ianpptrhoexidmiaettelfyor3.th5e%bCecae)dwiangsabdidredsd 0to
approimately 3%, sighy above the minimums recommended for quail (23%) and mallard (275%) 8).
Offpring limestone.
received
basal
diet
withou
test
substance
and
without
the
addition
of
5%
supplemental
veseiveWdaatneyr fwoarsmosufpmpeldiiecdatbiyotnheiotwhencocfdEdxsuonrtpihubntlisgc.watFeeredsuapnpdlyw.ateNreiwtehreer atnhalaydzueldtspenroirodoifcaflliynign accordance with Wilde Interiors, Lt. Standard Operating Procedures.
Dict Preparation the fina"lTedsitct.dieCtsonwterrole dpirtepaanreddcabcyhmoifxithneghPFrOStreinattoed dpirtiwxerethpatrewpaasreudsweedefkloywbeeegkilnyipnrgepoanraJtainounaroyf p2u3r,i2ty00o1ftahnedepsrtesseunbtsetdan1c0etahnedbiarrdespornesTeunctsedaays opfaersscpherwemeikl.lonDiaecttairvye cionngcreedniternatti(onpsmwear.e..adjDuestatieldsfoorf he weekly preparation oftest and control dicts ar shown n Appendix IL
Diet Sampling the traHtoemdadpigeetnscaintdyoofntehsamtepsltesufbostmantchee icnotnhteoldideitctwaosnvDal0uaeodfWbyeyeckoll1.ectSianmgpslixessawmeprleescofllreocmteadcfhroomf
"1037
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Wildlife International, Ltd.
Project Number 454-109
we
the 10
top middic and bottom dict samples wer as
of he Iet and cllected rom
righ scion of the mixing vesel. th fed osghs on Dy7ofWeek
Control 0 ass
and treatment sibility ofthe
ttesetsstubestarnoceuunddiectr,swchuentaesvacolnd,itidoinsn.g AWedekis n2,a4,,sa1m2p,l1e63wansd c2l0lfsthed cfsrtotmo tmheescsorntsrvoleiatnyd
fest concenstions. The i samples wee International Li. nalyice chemistry fit
sre ronan for alysis,
a
wns
mmedinely
to
the
Willie
Ansytica ithoa WifeTheInmcemtahtoidoauls,dLfio.r tahnedaneanlydseidso"fAnPaFlOytSicianlaMviiatnhodidckVewraisfibcaastieodnuoprontheetDheoeeliongaytidoenveolfopPeFdOS imnetAhvoidaonloDgiye"we(reWiinecornpoerramteadioinnato, Lthie.pPrreosejnctt Nsotu.dy4.54CT-h1e1s0)e.chTahnegeesmowdeirfeica(t1i)ontshefursom tohf chiitgehd psienrgfloermqaunacderulpiqlueidmcahsrsomdsettoecgtriaogn1hy wirtehdupcelepetqcunatdiraulpomlae.rimasisrdeeentieon,((H2P) LhOeMcSlMiSn)tronerofthtaen (i9n)etrhealssetafndaarsdihzoarttison(5p0romcedmur5e.1b0e0mcamus)e yan atpprocphrriomnateomgraapshtyacndoalrudmncoudxoptibe adlynstis,meand
Samples were exacted with methanol, vac Fire, died in than, nd thn ited in a
P50E:O5S0 mmeetthhaondooll:ogNyA.NOApumreethowdatfelrodwilcuhtaiotn isolpvreonvtidseodthiantAtphpeeynfdeillxwiItV,hiFnitghuercal1. ibrCatoinoncernanrgieoonfstohef
PFOS sing
in the standards Hewlt-acard
and exacts Model 1100
oHfigthhePesrafmoprlmeasncweerLeiqdueitdeCrmrionemdtobygrrpevherrse-phcase
HELCMSMS witha Pekin
TElhmemrasAsPIse3c0r00omMeatsrsvSapsecstqroimpepteedr owiptehratePderikninmlultirpleTruerabcotiloonnSmyornaiytofroinngso(urMceR.M)HPdeLtCectsieopnarratoidones
wee ited
achieved using a Keystone Betas Ci analytical witha Keystone Javelin Cs GuardCarcidge (20
cola mmx 2
(50 mmx mr LD)
2 mm LD, um parce size) Th instant pastes are
ummacized in Appendix IV, Table 1.
of
81C0al0ibrmatgionisLtandsatrodcskwesrleuptroenparoefd
in 50:50 methanol: NANOpurc water
PFOS in methanol. The calibration
by approprist dilutions
sandrds, ranging in
cpornocmeinnteanttipoenafkrormes0p.o0n0s10f0or10P0R.O0S100wamse.u.t.ilizewteoremoanistyozrePdFOiSthinacllhcsaalmibprlaetieotn., qTuhaleitsyanceoand,moasntd
Cingg
-_--
Wildlife International, Ltd.
Projoct Number454-109
Lsitndassagmrpelses.ioNnoeaqtueamtipotnswaswemraedegetnocqrutaendtifyusiPnOg Stheanptehsekbaasriesaofsipnodnisveisd!viesormserihcecosmppeocnteinvte IcV,oFnicgree2n.Ttohfrethaceotanrniiaortaintoosnn sofaPnOdrS.inAanvienxafmopdlesaomfpcleslwaas en cturemibsyprsesenstednin Atpheppeisnk asalsbaitno shieanadapdlscaarblsoliwnnearinreAgprpeessnidoinxcIaVt,iFoing.uresTy3piacnadl4,fornescpehcrtiovmeltyo.gaEmxasmpflesloowfeaqnudatihoignhs used incalulras rtesoenntesdinAppendIiVx,Table2.
he prohduectionfsttrhumennetcltiiotnovfodleuemeeti(o1n0.(0LLOLD))aodr tthheseloawneasltysseasnwdaesd1c0on0cpengtroa-t<ioonlu1,m0,0ucgalal1. ed f1o0m pProBduLct.ofthTehelomweetsthocdallimiitborfoqsuaacanrtal(yzeLd O(o0r0G0h1o)msegana1ly)sesanwdashe2.a0v0eprpmi.ticonltelooaefrhtdhee mati lanksam(2p000lLKe).Examples ofclclaartriesoenndin Append IV,Tale2.
Along inefrencss.
with No
he sample. analyses, seven inrfernces were observed
max. ot or
banks above
were analyzed to determine the LOG durin the sample
possible nly
(Appendix IV, Table 3). Figures.
A typical ion chromatogram of a mati blank is presented in Appendie Tv.
sampleAsvitaondieeemisnamepltehse wmreaenfpororcieraa trheec.ovleoryw.inTghelevferlsicatnidoanmlzevsedls cwoenrceur5r.e0n0t,ly50wi0t,h ahned T2h0e0 5fprimc.tifornsalmepvellisnwgeien5c.0r0, W25e.c0,k a,ndDa1y000 pWmeeka.1,foDaysa7m:pWliencgki2,evDaaly 0W;eaenkd W4,ecDke,y0,DaTh0ye, Wfeoerkt16i,sDalyv0l:sanwderWeec5.k0020n,dDa2y50.. pTphmemae.thoordsaimlpldiengmeevslprsoceWdeuerakl 5c,oDavye0sWofe1e011%2,,D1e0y10%,, (1A0p25p4e,nd1i0x0I%V,anTdab9l9e%3,).eSsampplee,meafsourrtehde s5o0n0c,e2n5at0i,on$s00w,e1e00n,tncd2o0m0sppmfora.hefoeraifnetpiroonceldeuvrealls recoerc. typical ion chromatogramof a tri rico s presen n Append IV, Fire6,
Crinsg
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Wildlife International, Ltd.
Project Number 454-109
6Study Phases
"The primary phases ofthe study andthiapproximate durations were: \ Aclimation 3 weeks. 2 Prephotostimuation- 10 weeks. 3. Preys lying(vith hotostimla0iwoens). 4 Egglaying -Approximately 10wecks 5. Postadul termination (nal incubation, hatching, and 14-day offspring rearing period-) 5
wks.
Housing and Environmental Conditions Housing and husbandry practices were conducted 5038 (0adhere10 the guidelines established by
{he Nasional Research Council 5). The adult birds were housed indoors in baterie of ens manufac ured by Safeguard Products, Inc. (Model No. $355), measuring approximately 75 X 90 X 45 cm high. "The pens were consirueted of vinylcosted wire mesh. A diagram of the fest layout is reseed in Appendix V.
Each pen was equipped with a feed trough. Weekly, sufficient feed for the feeding period was placed in the troughfo cach pen and presented {0th birds. Duringth fecding period addional feed was weighed nd added othe troughs 3s needed. Water was supplied by nipple-ype waters.
Only birds associated with tis study were maintained in the study room in order 0 avoid encesive disturbances. Theverag temperature in th adult mallard study room during the corseofthe Lest was 217C & 11C (SD) witha average relative humidityof40% & 19% (SD). Theai handling stem in the study room was designed 0 vent up {0 1S room sie volumes every hour and replace them with fresh ir
"The photoperiod in the adult mallard room ss maintained by a tim clock. The photoperiod during acclimation and the frst 10 weeks of the test was cight hours or es of ightprday. The shotoperiod was increased 0 17 hours of ight pe dy at he begining of Week 111 induce gg lying. and was maintained at that length ni he adult birds were uthanized. Throughout the es he birds received ameanof approximately 188 lux (~ 17. candies)ofluminaton provided by fluorescent lights {hat closely approximated noon-day slight
61.920
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Wildlife International, Ltd.
Project Number 454-109
ne Observations
"he est bis wer acclimated 0 the ails and stdy pens fo approximately3 weeks prio {niiation of the test. During acclimation, all birds wers observed daly. Birds exhibiting abnormal behaordvbiilaotirng physical juris werenotsed forthe test. Din te study, al adult birds were observed daily or signoftxiity or abrormal behavior. Additionally,al offspringwerebserved daily {rom htching uni 14daysofage. Arecwasomairntaidnedofsll mortalities and cial observations.
Adult Body Weight and Feed Consumption Adboudywleigthts wer measured at est nition, on Weeks 2, 4, 6, , and st adult emit.
Bods weights wre not measred din cg lyin becauseofthe posse adver fc handing may haovnceg prodution.
Food consumption for cach pen was measured weekly throughout te test. Feed consompion was determine by weighing th fishy filled fede on Day 0, recordin the amount ofany addins! dik added during the week, and weghing hefod and remaining fedt the end ofth fing period Dey 7). The amount of feed wasted by the birds was no quantified, sin the wasted feod was normaly catcred and mixed with water and xcrt. Therefore, ed consumption is pest3e a estimate of tal fed consmpion.
Adult Blood Collection Four addons paisofadult bids were ade 1cach test elt be mintind fr collection of
blood samples during the si. These pais received th same diet and wee handled in the same manner a3 other pais in the tet group. However, because of th. patel stress of the blood collection procedures, dat from th four adional pais ers no sed 0 value tment lated fc. Blood samples were collected fom birds in thee designated pairs, when possible, at five week inirvals hroughout the ts. A he time of adult termination, blood samples wee cllted from all surviving birds, when possible prior to. euthanasia. All blood samples were separated into serum and hemacyte/pltlet, sored ose and sipped othe spanar or sponsor's designate fr possible analysis
Adult Necropsy and Tissue Collection When the remaining bis fom 15020pm . cament group were euthanized a the end of the
Week 4, no necrosis wer performed. All ther dat bis tha died o were euthanized during the
.
cians
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Wildlife International, Ltd.
Project Number 454-109
"1s courseofthe study wee subjected 10 gross necropsy. Ath conclusion of the exposure period, al surviving adult bids wer euthanized by cervical dislocation, necropied, ad disposedofby incineration. At the time of necropsy, tissues wee collected fo histopahologial examination and posible analyses. When available, samplesofall blader, liver, proventriculus, Kidneys, rin, gonads, burs of Fabricius and adipose tissue were fixed in 10% buffered formalin and shipped to EPL in Herndon, VA for histopathology. When availble, samples of bile, iver, proventriculus, Kidneys, bran, reproductive organs and adipose tissue were sored frozen and shipped to the Sponsor for possible analysis. Fecal sampleswrealo collected, when avaiable, rom he ower digestive tract and sored frozenfo possible analysis. Eas Collection and Storage
Eggs wer collected daly fom all pens, when availble, gs to be incubated were washed 0 reduce the possibility of pathogen contamination before storing hem in the cod room. Eggs were washed in a commercial gg washer (Kubl Figg Washer) with a chlorine based detergent (Kubl Super CD). Watinetrhe washer was warmedto approximately 45C. The eggs wer placed n the wash water and soaked fo approximately 15 seconds. The washer' circlation mor was hen fumed on for approximately three minutes. The gs were removed from the washer, allowed col 0 approximately oom temperaturean rinsed wit fresh wt, Theeggs were then stored in codoom ulincubation. "The cold room was maintined at 3 mean temperature of 13.5C + 02C (SD) with a mean relative humidityofapproximately 69% 1 6% (SD). Groups of eggs were denied by a alphabetic lot code. loge id ina weekly interval were considered a oe ot.
Canding and Incubation A he nd ofthe weekly interval, all eggs wer removed rom th cold room, counted and ges
selected by indiscriminate draw for egg shel thickness measurement. The remaining eggs were candied With a Speed King (Model No. 32) cgg<andiing lamp to detct gg, shell crack or abnormal cggs Cracked or anoml ges were recorded snd discarded
A eggs ot discarded or sed for eg shel thickness measurements wee placed in a Petrsime Incubator (Model No. SP20). In the incubator the temperature was maintained at an average 375C + 00C (SD) with an average wet bl temperature of 30.5 + 0.35C (5D) (rlative humidity of approximately 60%). The incubator was cupped with pulsator fn and blades that produced a mild
cope
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Wildlife International, Ltd.
Prject Number 454-109
"0breathing air movement designed to climinate inacabint temperature and humidity aristion during incubation. In order to prevent adhesion of th embryo to th shell membrane, the incubator was also equipped withan utomtic egg rotation devie, designed orotthae tegges from 50offofvertical in one diestion to 50 off ofvertical in the oppesite ircton (otal rc of rotation was 100) every two hours {brough Day 24 of incubation. Eggs were candied on Day 13 or 14 of incubation to determin embryo viability andonDay21 to determine embryo survival Hatching and Brooding.
On Day 24 of incubation, the eggs were placed i a Petrsime Hatcher (Model No. SP-6H) and allowed to hatch, Pedigree baskets consiucted of gabvanized ice wire mesh were used 10 keep hatchlings separated by areal pen of origin. Eggs were no rotated in the haher. The average {emperstur nthe hting compartment was 37.24+C 00C (SD) and the average wet bulb empeature was raised to 3.3% 02C (SD) (lative humidityofaproximatly 80%).
A teins, unbached eg, an cg shels wereremovedfrom thehatche on Day 27or 28 of incubation. The group body weightofthe surviving hatchlings by pen was determined. Hatchlings were wing banded fo identification by pen of origin and then routinely housed according 0 the appropriate: parental concentration grouping in brooding pes unl14 daysof ag. The hatchlings were fd unveted diet without th addition of % supplemental limestone At 14 days ofag, he average body weight by parental pen of al surviving ducklings was determined. The ducklings were cuthanized with carbon dioxideand disposedof by incineration.
Hatching were house in bateris of broding pens manufactured by Safeguard Products, Ine. Each pen measured approximately 62 X 92 X 25. cm high. The walls, floors an ceilings of cach pen were consinuetedofvinyl.costed wite mesh. Thermostats n the brooding compartment of ach pn were eto maitemnperatsure of approximately 38C rom thetime ofaching uni the iswer ive 0 seven days of age, when the temperature was adjusted maintain a temperature of approximately 29C: The average ambient room temperature was 24.2C + 12C (SD) with a average relative humidity of 77% 15% (SD). The photoperiod for the hachlings was mainaind ba tyim clock a 16 hours of ight
perday.
:
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Wildlife International, Ltd.
Project Number454-109
"a OffspringBloodaad Tissue Colction
Pro to euthanasiaofthe Last groupof offspring (Lo 1), blood samples wer collected from 10 offspring in both the control group and 10 ppm a. reament group. All blood samples were separted into serum and. hemacytesplatelt, stored frozen and shipped to the sponsor for possible analyses. Additionally, iss fom the 10 offping in cach group were collected for histopathlogical examination. When avilable, sumplsofgall Madde, ver, rovnticuls, Kidneys, rin, gonads, bursa ofFabricius and adipose sue wee fixed in 10% bur formalin and sipped 0 EPL in Herndon, VA for histopathology. When avilable, samplesof bile, iver, proventriculus, Kidneys, brain, reproductive organs and adipose issue wee tore fozen and sipped othe spon for possible analyses. Fecal samples wee ais cllcted from pans below the flo ofbroaders housing Lot J offspring and sored froze for possible analysis. A single composite sample was collectedforthecontrol group and a single compost sample was colle forthe 10 ppm ai. resent group.
Ege Shell Thickness Measurements and Egg Components Collection Weekly throughout he cg lyin period on egg ws cllted, when svalble, rom each ofthe
{est pens. Eggs were collcted fom odd numbered pens during odd numbered weeks (1.3.3, tc) and rom the eve numbered pons durin the ven numbered wecks (246, ic). The eggs from weeks 1,2 and 10 were separated into alum, yolk, shell and membrane and ll components were toed fozen Eggs collected during weeks 3-9 wer open t ths wis snd thei contents removed. The eg contents were separate nt bmn a yolk and the yolk was tore faze. All og samples were tmmfered 10th Sponsor fr possible analysis. Th shells fom Wesks 3 though 9 were allowed to idy for at east one weeka room temperature ror fo being measure for thickness. The average thickness of the ied shel plu the membrane was determinedbymesuring five points round he waist of th egg using amicrometer. Measurements were made 0h nest 0.002
Statistical Analyses Upon compleon of the ts, an analysis of ariance (ANOVA) was pertonmed to determine
utisiclly significant iferences between groups. Duets multiple comparison procedure (67) was used to compar the thre samen means with th conol group mean and assess the stata ignifcance ofthe observed differences. Sample units wer the individual pens wii cach experimental aru, except adult body weighs where the sample unit was he individu bir. Percentage data were examined using Dunnets method folowing csi square oot tstomation (se Appendix VI fox
61024
Wildlife International, Ltd.
ProjectNumber454-109
a. reproductive parameter). With the exception of iver weight, statistical analysis of the data was performed using Avian Reproduction Data System (ARDS) Softwar, a validated software package developed by Wilde Intemational, Ld. (3). Liver weights were compared by Student's cst using. Microsoft Excel. Bach ofthefollowingparameters was analyzed sitistcally:
1. AdaualdtuBltodtyerWmeiniagtihotn.- InSdtiavtiidstuiaclalbocdoympwaeriigshotnswawsermeeamsudreed batettweseteinnittihaeticono,ntWoleger2ok,vp4s,an6,d8caancdh ratment grsotucapch weighinginterval by sx.
2.
ACdexuoalmtniFnseeedudbmyppentweieoklny
d-uFreiend
tchontseustm.ptSitoantisetxicparlescsoemdpaarsisgornasmsweorfe
fmeaeddepebretbiwrede
per day
n the con
was.
trol
and cach atm group.
3. EM ggsa Laix dofimuLam id - The number of eggs laid per female dividedby the largest number
ofcgas ad by any one female. This ransormation was usd 0 conver he mamberofgs aid 10percentile value ess tho eaqunal 1100.
he umber of ces sid, perpen. 4. EC ggsracofk Egge sLad id - The numberof eggs determined by candling to be cracked divided by
5. VE ibaybtlhm ee nbumrobofy gefEsrgo gsseSs t,cpter- pTehne number of viable embryos at the Day 14 candlingwasdivided 6. Live3:WoskEmbryosofViableEmbryos - The number of live embryos at the Day 21 canding
was divided by th mameofible embryeorspe,n.
7. Hatchlingsof3-Week Embryos -The numberof hatchlingsremovedfromthe hatcherwasdivided
bythe numbofeivr 3-veek embryos,pepen.
8. 1S 4nu-mDu baeyrOor lfdhav tchli ingsv posfo Hwaetcr ckh,lbis yngpsen.- The number of 14-day old survivors was divided by the 5. HaleholfEiggnsgSest- The number of hatchlings was divided by the number of eggs set per
wes, by pen. 10. 1S4:DuayOrM vivofEogesSref-The number of 1-day old survivors was divided by the
numberofeggs set per week, by pen. 11. Hatchling of Maximum Set - The number of hatchlings per female divided by the largest number
of eggs set from any one female This transformation was used to convert the number of batch lingstoa percsnile value quatoo ss than 10,
cnigs
Wildlife International, Ltd.
Project Number 454-109
"a
[I ------ 12. 1S 4-Dau yOlr d viov fMo axir mumSs et - The number of 14-day old survivorsper pen divided by
13. EggShellThickness - The average egg shell thickness of indiscriminately selected eggs per pen
was measured
14.
OsfufrsvpwBerioridevymneWoagesriu'grshesdt
-
by
The group
parentalpe
body weights
group.
of
surviving
hatchlings
and
14-day
old
15. LiverWeighs Individual liver weights were measured at adult terminationand at termination of
elcted Lot offspring. Statistical comparisonswer madbeysex between the controlgroupand
10ppm i. group.
RESULTS AND DISCUSSION Mature malads received PFOS at nominal diay conceatrations of 10, 50, and 150ppm ai. Duc 0 extensive overt signsofoxiianttrestmentrelated moraliis inthe 50 and 150ppm a. caument sroups, the experimental design was mii. Adults in the 50 ppm 8. test concentrations were euthanized th coafWedck ofth test, Thedistary concentrationfo the 150 ppm .. ramen evel was reduced 20 ppm a. of PEOS atthe beginning of Week 3 ofth test. The 20 ppm a. level was terminated at the begining of Week $ of the test due to the limited recovery of bids at his level. "Therefore, the test was completed with th control group and 10 pp ai. test concentrations any. The control group was fed dit comparable to the treatment groups, but without the addition of the est substance.
Diet Analytica Results None of the contol samples showed any indication of the presence of he test substance ofofthe
presence of co-luing substances atth characteristic retention time of the tet substance. Diet samples were collected fom th 10,50 an 150 ppm i. est concnrtons an Week 1, Day 0, andwere analyzed 10 evaluat th homageneity of th tst substance in th diet and to verify test substance conenieations `Means and sandard deviations forthe thre fest concenirations wee 9.75 0.313 pp a, 50. + 0981 Pom ai. and 147+ 431 ppm ai. respectively, The coeficients of variation were 3.21%, 196% and 2.93%, respectively. These values represented 98%, 100%, and 98% of nominal concentrations (Appendix IV, Table 4)
Cong
Wildlife International, Ltd.
Project Number454-109
6. Samples collcted throughout he et to very et substances concentrationsfo th 10, 20,50, and 150ppm .. dit hadmeansandstandarddeviationsof 102.2 0.353 ppm i, 208214pom a, 509 200 ppm a, and 161 15.6ppm ai, espesively. The contofvariation were 5.4%, 103%, 401% and 966%, respectively. These values rerescnted 102, 104, 102, nd. 108% of nominal conceneations (Appendix IV, Table 5). Anaisofdit samples collected fom Feder fe being held a ambient temperature for 7 days average 100%, 101% and 97%ofth Dy0 values forthe 10,50snd 150 pp i. est concertos, respectively (Appendix IV, Table 6). A typical on chromatogram ofa est sample is shown in Appendix 1V, Figure 7. Mean weekly measured concenaton fo cach est sroupare presented in Table | Mortaites While ther was on incidental mari in the conrl group, there wre no morales i the 10 pom ai. teament group. Thise incidental mortals and ten rcamenteated morales occured in the 50 ppm ai. trestment group and four treatmentrelated morales occured in the 150 pom ai. tresiment group.
"Th single incidental morality in the contol group waamlse (Pen210) ht died during Week 15 ofthe test. Gross necropsy of he drake reveled foo lesionons both fect. Ttemally, emorehges were note in he myocardium, the sbdomin) sir sacs were cloudy nd he spleen was enlrged and motd Additional, te live was fem, moti and pale the Kidneys wee slighty pale and 3 small aman of lear fluid ws noted inthe abominal cavity. Necropsy ofthe bird's pena at terminationofth sult pornofthe study revealed regressed ovary, but ws herve unremarkable
Thee incidental and tn este elated morals occured i the 0 ppm a. reatment group and our sament lated morale occured in he 150 ppm a. raiment group. Necropsy findings for the treatment related mortalities in the 0 and 150 ppm a. tet concenations were similar. Common findings included tin or emaciated condition witha loss of muscle mas and prominent kee, small spleen and ple Kidneys. Allremainingbids t he 150 pm .. et concentration wer chaized at the egoningofWeek 5. Due to th shortened natureof th exposure period, necrosiswereot performed A remaining birds at the 50 ppm 3. est conseniration were cthanized at the beginning of Week of the test, snd aecropsics were performed following cuthanasia. Necropsy findings are summarized in Tale
chron?
Wildlife International, Ltd.
Project Number454-109
"ae Clnist Observations
No overt signs of toxicity were observed a the 10 ppm i.test concern. With the exception of foot lesion and occasional lameness, al birds in th control group and 10 ppma. raiment group ere normal in sppecance and behavior throughout the es.
Inthe 150ppm a. and SO pm. tresment groups, igs ftoxicity werefirstnedat thecndof Weck 2. One male ithe50ppm a. treatment group and sera bids in the 150pm .. reament group exhibited signs of toxicity induced by the sess of handing during the Week 2 body weigh measurements. In several case, binds convulsed during body weight measurements. Siogf nxicisty in both the 50 and 150ppm ai. rete groups continued be absrved unl the birds were uthanized. Clinical signs of toxicity observed included depression, reduce reaction o extemal simul (sound and movement), wing droop, lossof coordination, rufed appear, hin appearance, lower limb weskacss, J p--
Adult TBhoedryeWweeirgehtno apparent restment lated ffcts upon adult body weight at the 10 ppm ai. test concentraatnidoan,y ifnces between th contol group and the 10 ppm a. Lest concenraton were not statistically significant at any ofthe body weight vals, However, there wee marked treatment related duction n mean bodyweight bt the $0 ad 150ppm. st concertos.
With he exception ofthe mean male body weights the Week2 interval, al differences between the contol groupand ids in the 50 ppm a. reatment group were saisticaly significant tp <0.01. At he 150 ppm i. test concentration, mean male and female body weghts wer snstcally diffrent from the control groupat p<0.01 at bth the Wee2k and Weck4 ody weight mera. Between he Week 2 and Weck 4 body weight intervals, 3 weight gain was noted for birds at the 150 ppm a. test concenration. However, he weight gin observed was a reflection of the reduction ofhe concentration of PROS in the diet 10 20 pp a. during Wesk 3 of th ts. Mean body weight measurements ae presented in Table 3. and Figures 1 and 2. Individual body weight measurements re presencd in Appendix VI
21028
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Wildlife International, Ltd.
Project Number 456-109
x. Adult Fed Consumption
"There were no apparent treatment lated effects upon fed consumption at he10or 50 pom 3. Let concentions. No satisticly significant differences bevweca the contol group and 10 or 50 pom a1 eatment grwoerupsersved atanyofthefedconsumption nerves. Howeve, resent ested reductions in fed consumpion were observed atthe 150 ppm 4. st cooencation during Weeks 1 and 2. The reductions were statically significant st p < 0.05 during Week 1 and stp< 0.1 during Weck 2. "The itary consenaion os he 150 pn ..restmen group was duced 120 ppm. during Week3 ofthe test. Ther was stitial significant p< 0.01) increas i sed consumpion atthe 150 (20) pom a. test concentration during Week3s and 4 ofth test. Mean sd consumption messurements ae shown in Tab4laned Figure 3. Feed consumption measubyrpecnamre pnrestentsed in Appendix VII.
Adult Necropsy and Liver Weights AN suiving adls in the contol grop and the 10 ppm .. eatment group were subjected 0
gros necropsy following adult termination. When compared10the contol group, ther was sn increased incidence ofmales with smal tse (spprosimate lengt<h 3.5 cmint)he 10ppm a. rcatment group or whicreatment lt affect coud notbeprecluded. Two maleinthecontol groupwer noted with smal testes, compared to seven ale in the 10 ppm .. group. All abe findings were considered unrelated to resment, Necropsy findarirenpogrtsed in Table5andAppendix IX.
When compare to th control group, thre wers noGeatment elated efficts pon the mean weighs of dul livers a the 10 ppm a. tsk concetation. Any difrnce between the 10 ppm a. seamen group and the control group was not sttsically significant. Mean adult liver weights re presented in Table , whit individual aul iver weiargphrestentsed in Appendix X.
Histopathlagy No lesions considered relied ots article (FOS) administration were noted nth iver, kidney,
adipose sue, brain, and proventriculus of aul snd offing males and females, bursa of Fabricius in oping males and femals, ovaries of alt and ofsring fas and ests of mal offspring. The increase in the numberofadult mals in th 10ppm . rated group ith duced testicular size (vithor without a corresponding micoscapc observation, Suggests tat PEOS may have accelerated carly post reproductive phase regression, a normal physiological phenomenon, Any polenta estatile eft on he bursa of Fabricius in adult males and females could not be determined since samples were rot
1029
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Wildlife International, Ltd.
Project Number454-109
2%submittedfor evaluation. Inaddition,any potential est-aricl effects onthe gallbladder rom adults and offspring could otbedefinitively determined due (0th extensive autolysis that precluded any accurate diagnosis. The fewothe changes in various tissuesofadult ado offspring mallards from control and 10 pm ai. treated groups were considered incidental and related otst ale (PFOS) dinisttion. "The full istopathology report is provided in Appendix XL
Adult Liver and Sera Adult blo samples (botre blood cells and srs) collected through Week 1S ofthe test were
analyzed by Exygen Research. The realsfo analysis of samples collected for this study ae reported {together with resuls for the nother bobwhit study ru concurrently with the same test concentrations. Summary rsuls of blood analyses performed for this study ar presented in Exygen report Tables X rough XXIV located in Appendix XIIA The fll report for analysis of blood samples by Exygen Research is located under Exygen Study Number 023066. "Exrocion of Potassium Perluorooctanesufonat fom Red Blood Cels and Serum for Analysis Using HPLC:ElctrosprayMass Spectrom"
Bloodand iver samples collected at the fime of adult trinaton or the control group and 10 ppm. a. test concentration were shipped to 3M Environmental Laboratory or analysis. Summary results of rn and ver analyses conducted by 3M Environmental Laboratoryar presented in SM report Tales 1. 2 and 10 trough 15 located in Appendix XIIB. The fll report fr analysisofsea and liver samples collected tthe time of adult termination i located in 3M Environmental Laboratory report EO1-1256 "Analytical Phase Report for POS: A Reprodution Study with the Mallar".
Reproductive Results There were no treatmentsated effets upon reproductive performance i he 10 ppm i. est
concentration as compared to the controls. When compared to th contol group, there were 0 aatistically significant differences in any ofthe reproductive parameters measured in the 10 ppm a. es. concentration. Summarics of th reproductive data arc prcscoted in Tables 7and 8, and in Figu4raensd 5. Reproductive parameters by pen ae presienAnpptenedidx XII and XIV.
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Wildlife International, Ltd.
Project Number 454-109
ae Egg Shell Thickness
her were no apparent reamen reltd fects up eg shell thickness at the 10 ppm i. tt concnraon, When compared 0 he contol group, there wee no atisicaly significant diffrence in egg shell thickness i the 10 ppm .. eament group. Eg shell thickness dota are prsented in Table 9 and Appendix XV,
Offspring Body Weights "Ther wer no sper resent reed ffs upon opin body weight the 10 ppm. fst
choencbeontdryatwieoin.ghtWhofenhactcohmlpiangrsedorto14t-hedacyonotlrdolsugrrvoiuvpo,rsthreroemwetrhee n[0o pstamtisatsi,calrlyessiegnnifticranot d.iffeOrfefnscpersinign ody weigh dota re presented in Tale 1,ad Appendices XV1 and XVII.
Offspring Liver Weights "There wer no rcatmentelated effect upon th mean weghts of offspring livers at the 10 ppm
a. et concentration. Any diffrence betwee the 10pm .. resmens group and th contrl group was so sical signifan. Mean opin fr weights are presented in Tale 6, while individal opin liver weghs ae presented in Appendix XVI.
Offspring Obsevacons There were ncdenal ahsrvaton of eck curl, weakened condition ad. foto, lesions and
ssid amencs, for offspring in both he convo group and 10 ppm a, esmens group. A he 10 om a. test concenraio, a general servation was made ta he oping fom Lots C andD were too, with light reduced eston to external simul, None ofthe offspring intheLosproceeding or following Lots C and D were neead lggih or wila reduced reson to xtra tml. Du to {he slated tr ofthe lincal sgn of oii noe, hes servations wer ot consid rcment rete. All ther ofpring were normal in ppeaanc and ehavio hroughout he st.
Ofspring Liver and Sera Blood and vr sample collie at temofoffspring termination for the control group and 10
gpm i. et concentration were sipped 0 Envircnmental Laboratoryforanalysis. Summary results ofsen and iver analyse conducted by 3M Evirnmena Labrtoy ar rset in 3M report Tables 1.280 10 trough 15 located in Appendix ITB. The fll rpor for analysisofse aod vr samples
@n10a
Wildlife International, Ltd.
Project Number 454-109
ne allcted tthe time ofjuvenile termination i lossted in Environmental Laboratory report E0I-1256. "Analytical hase Report for PEOS: A Reprodustion Std withthe Mallard"
concLusion There were no tetment lated morals, overt signs of txii, of treatmentltd effects upon body weigh, esd consumption or reproducive performance tthe 10 pm &.. est concentration. Howener there was an ness nidencofsll ses for male inthe 10 ppm a. et concttion What my hav been elt to eaten, There wereGestment elated moraites, ovr sige ofeniety and rcament lated effects pon adult body weght at the 50 and 150 ppm a. est concentrations. Thre were aso treatment ltd effets ed consumption the 150 pp. test concen, Th 50 and 150 pm .. est concentrations were terminated prio 10 he reproductive phase of he tes. Based on he effec observed at the 10ppm a test concentration, the mo-observedeffect concentration ox mallards exposed o PFOS in the diet was determined 0 be es than 10 ppm .., th lowest concentration tested during this su.
NLO3T
Wildlife International, Ltd.
.. REFERENCES
Projet Number 454-109
| USEbnSvd.ivriEosnaimvoeinnrtoanlEmP,reonHtteaaczltairPodrnoAtEgevecatnlicuoyan,tiOoAfngf:einccyo.WfiPdels1tf9ie8c2i.daenPdrPeosgAtrqiaucmaistd.iecWAassOshreigsnasgnmiiesonmnts,,DG.uCsi.udblsiencetiso,n.FI7F1-R4A, 2 OOrEgCanDiGzuaitdieolninfeofroErcTonoemiscoCtfoCohipeemirncaatlgiso.n aGnuiddeDleivneel2o0p6m.eaPta.rs.1984. Avian Reproduction Tes. 3 ARmeeprroidciacnivSeocSieutiysfowritTheAstviinag SapnedcisM.aterAiSalTs.M St1a9n8d6a.rd ES1t0a6n2d:a6r.d PrAoncniuiacle fBoorokCoofnAduSctTiMng
Standards. Vol. 11.04. PiadelphiPaA,. 15 pp. 4 Me&rCoc.Iknc. 1991. TheMerck VeterinaryMaral. Mer&cCko. Rahway, NI. 1932 pp. 5 NWaatsihoinnagltonR,esDeCa.rcNhatCioounnacliAl.cade19m9y6.PresG.uid1e23foppr. the Care an UseofLaboratory Animals 6 wDiulnhnetto,ntCoW.. Jo1u9.55.AmeAr.Mtualtsi,plAesCoocmpa30r:is1o0n9s6-P113r1 ocfoerCdomuparrieng Several Treatments
7 Duonnnett, CW. 1964. New Tablesfor Multiple ComparwiithsaCoonntrosl. Biometri2c0s: 4828 Wide Internationa, Lud. 1994.Avian ReprDatoaSydsieum(AcRDSi)Veorsionn 2.
"1033
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Table 2 fSoummamaMraylolfarGdrRoespsroPdatuhcotlioognicSatludOybsweirtvhatPiFonOsS Adult Birds FoundDeadEuthanized duringtheTest
I MiI les LFemaL les La
Opmai) mai ppmai
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2
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o
1
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1
0
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1
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1
0
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0
1
o
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3
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[rr --------.
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'
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Not remarable
0
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1
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3
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1
5
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1
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1
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1
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