Document 3Q205yVeB5D2j4be3QQ6pn6aE

ADIETARY LS0STUDYWITH THE NORTHERN BOBWHITE PPFFOOSS:: A DIETARY LCS0 STUDY WITH THE NORTHERN BOBWH1TE `WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL LLTTDD.. PPRROOJJEECCTT NNOO..:: 445544--110033 33MM LLAABB RREEQQUUEESSTT NNOO..:: U122772233 Sri 850 EGlPgFYiSnNaEmibsa 1T50s.2i300ns(4s) . UU..SS.. EEnnvviirroonnmmeennttaall PPrrootteeccttiioonn AAggeennccyy Series 850 - Ecological Effects Test Guidelines (draf0 OPPTS Number 850.2200 FFIIFFRRAA SSuubbddiivvisiisoinEoE,n, SSeeccttiioonn 7711--22 w (OOEECCDD GGuuiiddeelliinnee 220055 AUTHORS: San. Galigir AUTHORS: Sean P. Gallagher CCoouurrtmneeyy SS.. CCaasseeyy : JJoo~annnn BB.. BBeeaavveerrss `RRaayymmoonndd LL.. VVaannHHoovveeau. STUDY COMPLETION: Api 26,2000 SSTTUUDDYY IINNIITTIIAATTIIOONN:: AApprriill 2211,, 11999999 ; STUDY COMPLETION: April 26, 2000 SSUUBBMMITITTETDTETDOO 33MM CCoorrppoorraattiioonn Envios Labrsary Environmental Labora.toty 535 Bu Ave 935 Bush Avenue St Pu, Mmmm 55144 St. Paul, Minnesota 55144 WWililddlliiffee IInntteerrnnaattiioonnaall,, LLttdd.. EEa8$ass55tt99ooi88nn,,CCoM Mooamamvrrmymzileaenanrra~6d0DD220ir11iv66ve00e11 Pele (410) 822-8600 Page I of 62 22880000..00000011 Exhibit 2800 arm) wees 3MA00233956 WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDD 2. -2- PROJECT NO: 454-103 PROJECT NO.: 454-103 GGOOOODD LLAABBOORRAATTOORRYY PPRRAACCTTIICCEE CCOOMMPPLLIIAANNCCEE SSTTAATTEEMMEENNTT SSPPOONNSSOORR:: 33MM CCoorrppoorraattiioonn TTIITTLLEE:: PPRFOOSS:: AA DDiieettaarryy LLCCSS00 SSttudyy wwitihtthhe tNhoerNtohrteherrnn BBoobbwwhhiittee `WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL LLTTDD.. PPRROOJJEECCTT NNOO..:: 445544--110033 3M LABREQUESTNO: 02723 3M LAB REQUEST NO.: U2723. SSTTUUDDYY CCOOMMPPLLEETTIIOONN::.~ApArpirli12266,,22000000 the"TThhUiiSss.ssttEuuddyywnwaasscvcoonnddiuuccPttrereoddtieincnotcciooommnoppAllgiiaemannnccceeyev,wi4itt0hnhGCGoFotoRod.dLPalla~braotbr1oamt6to0orryyanPPrdraacc7tt5ii2cc,ee1SS7ttaaAnndduaagrruddssstaa1sp9pu8ub9bll;iiOsshhEeeddCbbDyy tPP1hrr9iei9nn2cUc)i.i;pSpl.laessnoEdonffvJGiaGropoooaonndmdeMLnLAataabbF!ooFrr,aaPrttooo5trr9eyycNtPPoirroahanccSttaiAicncgee,e,,nNc((oyOOt,CCifDD4iFEc0d/atGGCiDDoF((nR9SN22oP)).ar33t3228,,51EE06nn,0vvAiiagrrnroodinncmmu7eel9nnt2utt,rMaM1loo7nnPoorAggoudrrguaaucppsthhtioNNn1oo9..B89u44;r53,,cOuPPE,aarrCii1ssD0,, AA1uu99gg2uu)ss;tt a11n99d8844JwwaiipttahhntthMheeAffFooFlll,looww5ii9nnggNeeoxxhcceSepapttnii,oonm:Notification No. 3850, Agricultural Production Bureau, 10 charTaThchteeetretieszstatstusibuosbnwtsataannsccepeewrwafaossrnmnoeodttaccchhcaaroarracdctitenerrgiitzzeeoddi3inMnaSactccaconordrddaaarnndccOeewwiptihtehfnfuullPllrGGoLcLePPdcucoromempslapilaaiandncMceee;t;hhhooowdwesve,evaren,r,dtathhleel rcraaahcwwcaordardacdattaetnaarciazeararweteiibotbenheiwinGngLagPsm.mapieanrifntotaariminneeedddiaincnctothhreedin33gMM taorar3cchMhiivvSeetssa..nTdTahrhedetOteepssettsraustubinbsgsttaPannrcoceceiedissubrbeeiesningagndrreeMcehheaathrm.aoecdtstd..rairnzid~ziaeilandl accordance with GLP. was"TTnhhoeetssdttaeabtbieilrliimttyiyonoefdftithneeafctoecssottrssudubabsnstctaeanvnciceetahanGndodorreedffLeearrebenonccreeatssottaarnnydPdraaarrdcdutuinncddeeerSrctcoaonnnddidatirtidioosn.nssooffssttoorargee aattthhee teessttistitee: was not determined in accordance with Good Laboratory Practice Standards. SSTTUUDDYY DDIIRREECCTTOORR:: Ah ublfe, om A SSSeeeanaminorPP..BiGGoaallolllgaaiggshhteerr ' DATE Senior Biologist lle `SSPPOONNSSOORR''SS RREEPPRREESSEENNTTAATTIIVVEE done A Lowel Ve Susan A Beach oate_4/27/e2 22880000..00000022 33MMAA0000223333995577 WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LTO LTD .3- -3- PROJECTNO.: 454-103 PROJECT NO.: 454-103 QQUUAALLIITTYY AASSSSUURRAANNCCEE SSTTAATTEEMMEENNTT "TThhiissssttuuddyywwaasseexxaammiinneedd ffoorrcocommplpiliaanncceewwitithh GGoooodd LLaabboorraattoorryyPPraraccttiicceeStSatannddaarrddssaassppuubblliisshheedd by the U.S. Environmental Protection Agency, 40 CER Part 160 and 792, 17 August 1989; OECD bPryintchieplUes.oS.fEGnovoirdonLmabeonrtaatl oPrryoPtercactitoince,A(geOnCcDy,E/40GDCF(9R2)P3a2r,t E1n6v0iraonndme7n9t2,Mo17noAgruagpuhst N1o.98495;,OPEaCriD, P19ri9n2c)i;plaesndofJGapoaond MLAaFboFr,ato5r9y NPorahcStaicne,, (NOotCifDiEca/tGiDon (N9o2.) 3328,50E,nvAigrroincmuletnutraMloPnroogdruacpthioNnoB.u4r5c,asP,ad1s0, A1u9g9u2s);t a1n9d84,JaTpahnedMatAeFsoF,fa5l9laNuodihtSsaann, dNinostipfeicctaitioonnsaNnod.t3h8e5d0a,tAegthriactualtnuyraflinPdriondguscwteiornerBeuproeratue,d f1o0 tAheuSgtuustdy1D9i8r4e.ctTohreanddatLeasboofraatlloaruydMiatsnaangdeminesnptewcetiroenas sanfdoltlhoewsdates that any findings were reported to the Study Director and Laboratory Management were as follows: Activity ACTIVITY TT&eesstAtuSSaulubybstsittcaaannlcceeSaPPmrrpeeplp.i. ng & Analytical Sampling Maix Fortification Matrix Fortification FF&eeeeAddnaCClooynntsisuucmamlpptStiaioomnnpling & Analytical Sampling AAnDnaralalyfytttiiccRaaellpoDDraatttaa aanndd Draft Report DBBriiooallfootggyyRDeDapoatrtaat.aanndd Draft Report FinalReport Final Report DATECONDUCTED ~~ STUDYDDDAAITTREEECRRTEEOPPROORRMTTEAEDDNATTOOG::EMENT DATE CONDUCTED STUDY DIRECTOR MANAGEMENT AApprirlil2222,,11999999 AApprirlil2222,,11999999 AApprrlil2222,,11999999 AApprrlil2222,,11999999 AApprirlil2233,,11999999 AApprirlil2233,,11999999 April 27, 1099 April27,1999 AApprrlil2277,, 11999999 MMaayy 4,,11999999 1Juullyy77,,88,,99,, 11999999 JJuullyy9,, 11999999 JJuullyy l166,,11999999 AAuugguus~t2266,,2277,,3300,,3311,,11999999 AAuugguusstt3311,,11999999 AApprriill l177-1188,,22000000 AAppriflill188:,22000000 SSeepptteemmbberer1133,,11999999 AAprpidlll199,,22000000 SusaSn.ugCoolenman. Coleman SSSeeunnsiaioonrrLQQ.uuaCaloliitlteyymAAassnssuurraannccee RReepprreesseennttaattiivvee pate___ DATE 44-19-00 22880000..00000033 33MMAA0000223303995588 WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LL'TrD0 a-4- "RREEPPOORRTT AAPPPPRROOVVAALL PROJECTNO. 454103 PROJECT NO.: 454-I03 SSPPOONNSSOORR:: 33MM CCoorrppoorraattiioonn "TTIITTLLEE:: PPFFOOSS:: AADDieiettaarryyLLCCS500 SSttuuddyywwitithhtthhee NNoorrtthheerrnn BBoobbwwhihtiete: W `WIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL LLTTDD.. PPRROOJJEECCTT NNOO~:: 445544--110033 33MM LLAABB RREEQQUUEESSTT NNOO..: UU22772233 SSTTUUDDYYDDIIRREECCTTOORR:: Alert: nly dvds : Sean Gallagher bee Scan P. Gallagher I Date `SSeenniioorr BBiioollooggiisstt,, AAvviiaann TTooxxiicolloogy `CCHHEEMMIISSTTRRYYPPRRIENNCCIIPPAALLIINNVVEESSTTIIGGAATTOORR:: Rank. ViaHoveFni,D 4 LLU | Ph.D SScciieennttiisstt,, AAnnaallyyttiiccaall Chemistry BaeA200 RREEPPOORRTTAAPPPPRROOVVEEDDBBYY:: \ SEs, Ton Bs fot foe JJoo~a-n--BB.. BBeeaavveerrss " ' ~ ' DDaattee: DDiirreeccttoorr,, AAvviiaann TTooxxiiccoollooggyy ction, Pa) Willard B. Nixon, Ph.l~. MMaannaaggeerr,, AAnnaallyyttiiccaall CChheemmiis-t~rryy LA Date 22880000..00000044 aMAD0233959 3MA00233959 W WIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTD us. -5- PROJECT NO: 454-103 PROJECT NO.: 454-103 TABLEOF CONTENTS TABLE OF CONTENTS TITLE PAGE ................................................................................................................... Page I GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT... Pie 2 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT ............................. Page 2 `QUALITY ASSURANCE STATEMENT... 38 3 QUALITY ASSURANCE STATEMENT ........................................................................ Page 3 RREEPPOORRTTAAPPPPRROOVAVLA.L............................s..v.m..o..r.e..s..s.n..s..n............... : ......................-- ...................C . PagX e 4 T`ABT LEOA OFFCCB OONTNL ETNETNE ST.S........................................................................................-- ........... Page 5 T`abTlesaaannddbAAPl pPpEeBenIdCis cEeSs ............................o..o..v..v..r..r..n..e......................................................c.o.re.r.e.n.e...... PPAaGgEe 66 SUMMARY ..................................................................................................................... Page 7 INTRODUCTION ............................................................................................................ Page 8 OBJECTIVE .................................................................................................................... Page 8 MMAATTEERRIIAALLSSAANNDD MMEETTHHOODDSS ................c.e.r.e.r..s.e.e.r.e.s..e.s....................................S..--.................... Page 8 TestSUbSIRE...cc. FE Page 3 Test Substance................................................................................................................ Page 8 Treatment Group... res Treatment Groups .......................: ................................................................................... Page 9 Duration Ft Tes. BE Page 9 Duration of the Test ........................................................................................................ Page 9 TestBids. Page 9 Test Birds ....................................................................................................................... Page 9 Animal Dic... CII Page 10 Ammal Diet .................................................................................................................... Page 10 Dict Preparation III Page 10 Diet Preparation.............................................................................................................. Page 10 Dict Sampling TI Page 10 Diet Sampling ................................................. : ............................................................... Page 10 AnalyticalMethod CI rent Analytical Method ........................................................................................................... Page 11 `Housing andEnitoarmental Condions sss PAGE 13 Housing and Environmental Conditions ........................................................................... Page 13 Observations... ere Page 14 Observations ................................................................................................................... Page 14 AnimalBody WeightFeedConsumpion. ores PAGE 14 Animal Body Weights/Feed Consumption........................................................................ Page 14 GOSSNCCTODY orto PA8E 14 Gross Necropsy .............................................................................................................. Page 14 StatisticalAnalyses... Page 14 Statistical Analyses ......................................................................................................... Page 14 RESULTS... -- -- on RESULTS ............................... i ........................................................................................ PPaAgGeE1S5 Dict Aualyss SR Page 15 Diet Analysis .................................................................................................................. Page 15 Mortalites and Clic Observations. I ee 1s Mortalities and Clinical Observations .............................................................................. Page 15 Body Weight andFeed Consumpticn...... JT) Body Weight and Feed Consumption ............................................................................... Page 17 Gross Necropsy... = =] Gross Necropsy .............................................................................................................. Page 17 CONCLUSION. CONCLUSION -- i. .................................................................... 2 ........................................... PPaaggee 1185 REFERENCES ................................................................................................................ Page 19 22880000..00000055 M3MA0A000223333996600 W WIILLDDLLIIFF'EE IINNTTEERRNNAATTIIOONNAALL,, LL-TFDD "-66 - TTAABBLLEEcoOOnFtFiCCnuOOeNNdTT-EENNTTSS - continued - TABLES AND APPENDICES TABLES AND APPENDICES PROJECT NO: 454-103 PROJECT NO.: 454-103 TABLE 1: TABLE 1: TTAABBLLEE2:2: TABLES: TABLE 3: TABLE4: TABLE 4: TABLES: TABLE 5: CCSutumumudlyualwatitiitvvhee PMMFoOroStrat.al.lii.tt.yy.ff.rroomm 2a Norhem Northern BBoobb-- wwhhiittee AA-- ccuuttse DD-- iieetaarryy -- TTooxxiicciittyy A2 Study with PFOS ....................................................................................... Page 20 MMSetaeuadnnyBBwoiodtdhyyPWWFeeiOigg.hhtt.(g.g))ffrroomm aa NNoorsrtthheerornnBBomobbwwhmihtiteeAmAccuut--teeDDi--icettaa--ryy TTo_oxxiicci--ittyy 21 Study with PFOS ....................................................................................... Page 21 MM`Aeceauatnne FFDeeieeaddrCCyoonnTsosuuxmimpcpitttiiyooSnntu((gdb/yibwririddt/dhaayPy)F)OfrSroo.mm aa NNoorrtthheerrnn BBoobbwwhhiittee Page 23 Acute Dietary Toxicity Study with PFOS ................................................... Page 23 GG`Arrcoouuutepp DGGirrtooassrssyPPaTatothhxooillcookggiyicScaatlluOOdbybswsceirrvtvahattPiiooFnnOssSffr.roo.mm.aa.NN.o.orr.tth.heemr BBoobbwwhhiittee Pages Acute Dietary Toxicity Study with PFOS ................................................... Page 25 CNCourmthuuelrantimvBeoMbMowuorhtritatalelliitAyyc(u(aEEtsesttiiDmtmieaatttaeeriddyCCTuuomvmxuliuacltatetiyivvSeetuDDdooyssceW,,itmmhgg/P/kkFggO)S).ff.rr.oo.mm..aa........ Page 26 Northern Bobwhite Acute Dietary Toxicity Study with PFOS ..................... Page 26 APPENDIXE: APPENDIX I: CertioffAnialcysaist..e. Page27 Certificate of Analysis ......................................................................... Page 27 APPENDIX II: APPENDIX II: Dict Formulation. smmm----------ig 24 Diet Formulation ................................................................................. Page 28 AAPPPPEENDNIDXIlXIII:: AAnnaallyyttiiccaall MMeethotds ahannddoRREdessuulsltss....................: cn .......................................... PPaAgeG2E9 AAPPPPEENDNIDXIIIXVV:: Diet Diet Prepastin......... -- Preparation .................................................................................. 1 Page 43 APPENDIX V: APPENDIX V: CACucmuuutmleuaDltaiitcivtvaeerMMyoTrooarxtliaicltiitytyybbSyytPPueednny ffWrrooimm a a NNoorrtthheeFrrnnOBBSoob.bwwh.hi.ittee Pagedd Acute Dietary Toxicity Study with PFOS ............................................ Page 44 APPENDIX VI: II`nAncdduiitvveiiddDuuiaaelltBBarooyddyTyoWWxeieciiiggthhyttssSt((ug)d) yffrrwoiomtmhaaPNNFooOrrtSthheerrnn Bobwhite Bobwhite Page 45 Acute Dietary Toxicity Study with PFOS ............................................ Page 45 APPENDIX VII: SFNtoeuredtdyhCweoirnntshBuomPbpFwthOiiSotne(AgcbuitredDdiaeyt)arbyyTPoevnicfirtyom a P26 58 APPENDIX VII: Feed Consumption (g/bird/day) by Pen from a Northern Bobwhite Acute Dietary Toxicity Study with PFOS ................................................................................ Page 58 AAPPPPEENNDDIIXX VVIIILI:: CChhaannggsess 0to Protocol... Protocol ............................................................................ PPaaggee 6611 AAPPPPEENDNIDXIIXXX:: PersvolovendinntheeStuldy' Personnel Involved in the Study~ smn .......................................................... PBaAgeG6G2 22880000..00000066 3MMAA002032333996611 W WIlLLDDLLII FFEE IINNTTEERRNNAATTIIOONNAALL,, LL-TrDD 1. -7- PROJECT NO: 454-103 PROJECT NO.: 454-103 SSUUMMMMAARRYY SSPPOONNSSOORR:: 33MM CCoorrppoorraattiioonn TTEESSTT SSUUBBSSTTAANNCCEE:: PPFFOOSS. `WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL LLTTDD,. PPRROOJJEECCTT NNOO:.: 445544--110033 SSTTUUDDYY:: PPFFOOSS:: AADDiieettaarryyLLCCS5O0SSttuuddyywwiitthhtthheeNNoorrtthheerrnnBBoobbwwhiittee RREESSUULLTTSS:: ."?;hThee ddiieettaarryy LLCCSS00vavlaluueeffoorrnnoortlthheermn bboobbwwhhiittse eexxppoosseeddttoo PPFROOSS wweass ddeteermtineedttroobbmee22i2200nppppemmada..i.wwiitthh aa 9955%%ccoonnfidfencieinidnteterervvalanl oocff116e644 ppppmm a~ii..1to028.9p2ppp8mma9ai.i.. TThheessllooppee oofftthheeccoonnccecntnrattirona-treospno-nrseeccsuurprvvoeeuwwsaaess 77..000055a~nnddtthheecehih-sqiuarsovvaqalluueuewwaaass0r0..00e2233..TThhee nnoommorotratalliittyyceoonnceevnattrraattiioonn wwaass 7733..22ppppmm aai.i.. BBasaseedduuppoonnttrreeaattmmeennttrerellaatteedd mmorotratlailtityy,,sisgignnssooff ttooxxiicciittyyananddefefffeeccttssuuppoonnbbooddyy wweeiigghhtt ggaaiinnaatttthhee 114466ppppmmala.j.tetesstt `concentration, the no-observod-<ffectconcentration was73.2 ppm a. concentration, the no-observed-effect concentration was 73.2 ppm a;i. TTEESSTT DDAATTEESS:: HHaattcchh -- AApprriill !122,, 11999999 AAcccclliimmataitioonn--AApprriill 1122:-2222,, 11999999 EExxppeerriimmeennttaall SSttaarrtt --AApprriill 2222,, 11999999 EExxppeerriimmeennttaall TTeenrmmiinnaattiioon-n- MMaayy 1144,, 11999999 NNCOOOMNMCIIENNNAATLLRTTAEETSSITTONS: 0, 18.3, 36:6, 73.2, 146, 293, 586 and 1171ppmai CONCENTRATIONS: 0, 18.3, 36.6, 73.2, 146, 293, 586 and 1171 ppm a.i. TTEESSTTAANNIIMMAALLSS:: Norther Bobwhite (Colirus irginians) Northern Bobwhite (Colinus virginianus) AAGGEETTEESSTTAANNIIMMAALLSS:: 1100ddaayyssooffaaggeoaat teessttnitnsittiaitoionn. `SSOOUURRCCEETTEESSTT AANNIIMMAALLSS:: WW8i5ill9dl8liiCfeeoIInmntmteeemrranctaeitiooDnnraaillvLLedtd.. PPrroodduuccttiioonn FFlloocckk Easton, Maryland 21601 8598 Commerce Drive Easton, Maryland 21601 SSTTUUDDYY CCOOMMPPLLEETTIIOONN:: AApprriill 2266,,22000000 22880000..00000077 33MMAA0000223303996622 WILDLIFE INTERNATIONAL, Lo WILDLIFE INTERNATIONAL, LTD se -8- PROJECTNO. 454.103 PROJECT NO.: 454-103 INTRODUCTION INTP~ODUCTION his say was conduciad by Wide nemanal Lid for 3M Corpor at the Wid This study was conducted by Wildlife International Ltd. for 3M Corporation at the Wildlife oermtionl Ld. vin ilo ls in Eon, Mayland Tho nif postion ofts fs vac International Ltd. avian toxico!ogy facility in Easton, Maryland. The in-life portion of the test was ccoonndduucctteedd ffrroomm AApprriill 2222,, 11999999 ttoo MMaayy 1144,, 11999999.. RRaaww ddaattaa ggeenneerraatteeddaatt WWilidldlliiffee IInntteermnaattiioonnaall LLttdd.. Wide ematical Lid. si aanndd aacocoppyyofofththeefifinnaallrereppoorrttarareeffiilleedduunnddeerrPProrojjeecctt NNuummbbeerr 445544--110033iinnaracrchhiivveess llooccaatteeddoonntthhee Wildlife International Ltd. site. OOBBJJEECCTTIIVVEE he bjuive ofis study vas to caus the toxic of3 test substance the Norm Bobwhite The objective of this study was to evaluate the toxicity of a test substance to the Northern Bobwhite (Coli virgins) adsiscedthroughted fx ve ey An LCSO val il be calli, (Colinus virginianus) administered through the diet for five days. An LC50 value will be calculated, irpovite if possible. Themethods usedinconductMiMnAgtA TEsRTLtME d,SyAR AaNNrDDIbMMaA EsEeTTdL HHOuOpS DDonSSprosedursspc inteUS, EnviTrhonememntetahlodPsrucseod ninAgency Seis 0 ~ Bcloial Eos conducting this study are based upon Test ridlns OPPTS procedures specified in NuthemUbe.Sr. E15nv0i2ro2n0m0en(1ta),l Scion 712 of Protection Agency tSherieEsnv8i5c0n-meEnctollogPicraol tEoffnectsAgTeensyt'GsuiRdeegliinsetsstiOoPn.PTGSuiNdluimnebse,r P8e5s0t.i2c2id0e0 A(1s),seSencttionGui7d1e-l2ineosf, tFhIeFERnAviSruobndmiveinstiaolnPBro,teHcatzioanrdAEgvenacsyi'sonR;egWisitrfatieonand Aqui Guidelines, Organs 2; OECD Guideline Pesticide Assessment Guidelines, 2F0I5F,RAGuSuibdnivisfioornTeEs,tiHngazoafrdChEevmailcuaaltsio,nA: vWiainldlDifeiaynd Toi Aquatic sOrtgan(i3sm; san(d2)u; pOonECADSTGMuidSetlainneda2r0d5E,EGS7u.id8e7l,in"eStfoanrdTaredstiPngacoifsCfhoemiCcoanlsd,ucAtviinagnSDubieatcauryteTDoixciacirtyy oxi Tos with Avian Spc" 4). Test (3); and upon ASTM Standard E857-87, "Standard Practice for Conducting Subacute Dietary Toxicity Tests with Avian Species" (4). TTeessttSSuubbssttaannccee "hetest substance wasrcsivdfom 3M Corporation an Octobe 29, 1998 andwasassigned The test substance was received from 3M Corporation on October 29, 1998 and was assigned Wiis Itraional Lid. Hentiaton Number 4675 up rcp. Th est substance was white Wildlife International Ltd. Identification Number 4675 upon receipt. The test substance was white powder enedas; FC-95,Lo N21. Th reported pury of ettssa was 8.9%, with powder identified as: FC-95; Lot No.:217. The reported purity othe test substance was 98.9%, with an epirditae ofi20n08, Followingtest termination, hetest material ws rear, The rss an expiration date of 2008. Following test termination, the test material was reanalyzed. The results ofreanalysis dicate est sbsans pryof 049%. ltt oncntruions avebin adjoe0d of reanalysis indicate a test substance purity of 90.49%. All test concentrations have been adjusted to et the purty repre on he new Corlte of Analysis (Appendix. The tt substance is reflect the purity reported on the new Certificate of Analysis (Appendix I). The test substance was oro ndeaint conto stored under ambient conditions. Wile terns Ld. dentition mune 4526 spon ect. Tho tra Sandgara,n "TThheeinintteemmaallststaannddaarrddwwaassrerecceeiivveeddffrroomm 33MM CoCroproproartatiioonnoonn JJuullyy22,, 11999988aannddwwaassasassisiggnneedd. Wildlife hltemational Ltd. identification number 4526 upon receipt. The internal standard, a granular 22880000..00000088 awsoozsases 3MA00233963 WW ILILDDLILIFFEEIINNTTEERRNNAATTIIOONNAALL,, 0 LTD 5 -9- ROCTNO:dsl PROJECT NO.: 454-103 Number, 27619:97:2. The standardwas ford nde ambit contiins m`maatteerriiaall,, wwaass iiddeennttiiffiiecdd 2ass:: 11HH,, 11HH,, 22HH,, 22HH PPeerrfflluuoomrcooccttaannee SSuullffoonniiec AAcciidd,, CChheemmiiccaall AAbbssttrraacctt Number: 27619-97-2. The standard was stored under ambient conditions. TrTerea"atTtmmheentntetGsGrrocoouunpspisssted of agem seisofsve ts.conection and contol group. Thirty sortThebteostwcionesisctleidckosfwaegreoamsestirgicnesdertioestohef soevnernltegsrtocuopnacenndtraetinonnsorantdeabcoonbtrboligrocuhpi.cksTwheirrtey northern bobwhite chicks were assigned m the control group and ten northern bobwhite chicks were asasssiiggnneeddttoo eeaacchh oofftthhee ttrreeaattmmeenntt ggrroouuppss.. TThhee bbiirrddss wweerreesosorrtteeddbbyy wweiegighhtt,, tthheenn cchhoosseenn ndiscrminstlyfrom wii cach repressed weightclass fo placement focomoand iment indiscriminately from within each represented weight class for placement into control and treatment ggrroouuppss.. TThheebbiirrddsswwereree hhoouusseeddiinnbrboroooddiinnggpepeanss ccoonnttaaiinniinngg ffiivveecchhiicckkss eeaacchh.. NNomoimninaallddiieettaarryy cocenenions usd nti dy ers O, 18.3, 366, 7332, 146,293, 55and 117 parts pe ition concentrations used in this study were O, 18.3, 36.6, 73.2, 146, 293, 586 and 1171 parts per million activ ingprpmea. dofiPOnS.The teary seniins wessibleSed uponknw active ingredient (ppm a.i.) of PFOS. The dietary concentrations were established based upon known ttooxxiicciittyyddaattaaaannddiinnffoorrmmataitioonnssuupppplliieeddbbyytthheeSSppoonnssoorr.. Eachgroupwasfothesppropriate et ciel itforfivedays.Duringthe xposprod tecEonatcrholggrrooupupwaessfiedvtdheunaprparotpdrifatedt.est Foorlcloownitrnogltdeiet fiovredfiavye deaxyps.osDurpienrgitohdeaelxlpgorsuorpespweeriroed thveecnountronl grourbpasarelacieivketdfourntterreatedddayfse.edO. nFDolalyow5,ingfthoeffitvhe-sduaryvievxipnogsturreeatpmeerniotdnadllcgornoturposl iwcerse given untreated basal diet for three days. On Day 8, half of the surviving treatment and control birds `wweerreecueuththaanniizzeeddaannddlliivveerrttiissssuuee,, bblloooodd,, aannddbbiillee ssaammpplleesswweerreeccoolllleecctteeddfoforraannaallyyssiiss.. TThheereremmaiainniinngg `bbiirrddsswwerereefefeddbbaassaallrraattiioonnunutnitill DDaayy2222.. OOnnDDaayy 2222,,tthheesseebbiirrddsswweerreecuetuhthaanniizzeeddaannddalalssoossaammpplleedd ffoorrlilivveerrwweiegighhtt,, bblloooodd,,aannddbiblilee.. "Toprayphase ofthis et a the duis ers: DDuuratrioanooftftthiheeToTeesnstt The primary phases of this test and their durations were: 11.. AAccclcimlaitiomna-- t1100iddaoayynss.. 2 Epon5daays. 2. Exposure - 5 days. 3. Postexposurcshc3.r1a1t7idaoyns 3. Post-exposure observation - 3 or 17 days TTeessttBBiirrddss All sorter bobwiit (Calne virgins) wre 10 daysofageand appeartedob ingood All northern bobwhite (Colinus virginianus) were 10 days of age and appeared to be in good health at nioif thenet. The bids were obtainedfom Wife Inematinal Li. Prodction health at initiation of the test. The birds were obtained from Wildlife International Ltd. Production Fk,Eason, MDandwer ached on April 12,199. Bis ang inweight rom1810 23 grams Flock, Easton, MD and were hatched on April 12, 1999. Birds ranged in weight from 18 to 23 grams a cst ition.Thebids sedi his studywer immatarsad coud otbs ifrenatdby sex at test initiation. The birds used in this study were immature and could not be differentiated by sex. 22880000..00000099 awsoozsases 3MA00233964 W WIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDD 10- - 10 - PPRROOJJEECCTT NNOO..: 445544--110033 AANllibdirdsswwereereffrroommtthheesseammeebhaattcchh,, ppeenn--reraereadraaennddpphehneontoytpyipciaclalllyyininddisisttiinnggouiisshhaabbileeffrroommwwilildd bbiirrddss.. Al All bbiinrddsswwerereeacaccclliimmaatteedd fo to th the ccaaggiinngg and and ffaacciilietisesffrroommtthheeddaayyooffhhaattcchhuunntiil niniitiattioanootfftithheeoteensstt.. AnAinimmaall DDiieett ThTrhorouugghhoouuttaacccclliimmaattiioonnaannddtetessttiinnggalalll etesstt bbiirrddsswwerereeffoedd aaggaammeebibrirdd rraattiioonnffoorrmmuullataetdettdoo W Wiidldilfifee IInntteermnaattiioonnaall LLutbd'.'ss ssppeecciiffiiccaattiioonnss ((AAppppeennddiixx IIID).. TThhee cchhiicckkss wweerree ggiivveenn aa vviittaammiinn `ssuupppplleemmeenntt iinn tthheeiirr wwaatteerr ffrroomm tthhe ddaayy tthheeyy wweerree hhaattcchheedd uunnttiill tthhe iinniittiiaattiioonnoofftthhee teests.t. W Waatteerr ffrioomm ththeettoowwnnooffEEaassttoonn ppuubblliiccwwaatteerr ssuupppplIyy,,aannddfefeeeddwweerreepprrovoidevdaaidddilibbeiihmdummdduurriinnggacacclcilimmataitioonn aanndd tteessttiinngg..TThheebbiirrddss rreecceeiivveeddnnoo ffoorrmm ooffanatnitbibiioottiiccmmedeidciactaitioonndduurriinnggaacccclliimmaattiioonn oorrtteessttiinngg.. DDiisett PPrrseppaarraattiioonn "TThheetteessttsusubbssttaanncceewwaassmmixixeeddddiirrceecltlyyininttootthhee rraattiioonn.. MMiixxiinnggwwaassddoonneewwitithhaaHHoobbaarrttmmixixeerr (Model (Model NNuummbbeerr AASS220000TT),). All All ddiiectaarryy est test ccoonncceennttrraattiioonnss wweerree aaddjjuusstteedd ttoo 110000%% PPFOOSS bbaasseedd uuppoonn the the rreeppoorrtteedd ppuurriityyooftf hthee tteesstt ssuubbssttaannccee.. AAllll ddiieettaarryy ccoonncceennttrraattiioonnss aanndd tthhee LLCCS500 vvaalluuee aarree rreeppoorrtteedd aass ppppmm aai..i. iinn tthhee ddiieet.t. NNoommininaall ddiiectaarryy tteesstt ccoonncceennttrraattiioonnss uussedd iinn tthhiissssttuuddyywweerree 1188..33,, 3366..66,, 7733..22,, 114466,,229933,, 558866,, 11117711ppppmm a.a.i. A(AppppeenndidxIiIVV)x). DDieiett SSaammplpilinngg SSaammpplleessoofftthheetetessttddiieettsswweerreecoclollleecctteeddttoo vveerriiffyytthhee tteessttccooncnecnetnrtraattiioonnssaaddmimniinsistteerreeddaannddttoo ccoonfnifrirmmththeesastbaibliltityyaannddhhoommogoegneenietityyooffththee etessttsusbusbtsatanncceeiinnththeeddiicettss., HHoommogoengeiteyonoffetthhieetetesystt ssuubbssttaanncceeiinntthheedideiettwwaasseevvaalluuaatteeddbbyycoclollelcetcitinngg isixxssaammplpelessffrroommtthhee 1188..33ppppmm a.i.aai...i.,tteessttdidciettaanndd ssiixx ssaammpplleessffrroommtthhee 11117711 ppppmmaia..i. tteessttdidcitetaattpprrepearpataiorn oaonntDDiaayoy0n0.. HHomoomgoegnesnietityyssaammplpeless wweerree. coclolllesctteddfforommtthheettoopp,,mrindidddlleeaannddbobtotttoommoofftthhee clef~aannddririgghhttsescetctiioonnssoofftthheemmixixiinnggvevsessseell.. TThhee. hhoommooggeennceiittyy ssaammpplleess also also sseerrvveedd as as vveerriiffiiccaattiioonn ssaammplpelse.s. OOnnee vveerriiffiiccaattiioonn ssaammppllee wwaass ccoolllleecctteedd frfxoommtthheecocnotnrtroolldideiett and and two two vvereiriffiiccaattiioonnsasammplpleesswwerereecoclollleecctteeddfrfroommceaacchhreremmaianiniinngg ttrreeaattmmeenntt groupat preparoantDiaoy0n. Attheend oftheexposu priod(Dy5), nesamwaspcollleceted fgrroomutphaetcpornetpraorlaatinondtownoDsaaymp0l. esAwtethreeceonldlocifethdefreoxmpocsaucrehtpreeraiotdme(nDtagyro5u)p, otnoedestaemrpmilenewsatsabciollilteycotefd tfrhoemtetshtescubosnttraonlceanidn ttwheo dsiaemt pulnedserweterestccoolnldeicttieodnsf.romTheeachstatbreilaittmy esnatmpglroeuspwteoredectoelrlmecinteedsftarboimlityfeeodf rthemeatiensitngsiubnsttahnecefeiendetrhse adfiettrubnedienrgteasttamcobnideintitontse.stpTehen csotanbdiilittiyonssafmoprlefsivweedraeysc.olleScatemdplfreosm wfeereed remaining in the feeders after being at ambient test pen conditions for five days. Samples were ttrraannssffeerrrreeddiimmmemdeidaitaetellyyttoo WWiillddlifeeInIntteemrantaitoionnaallLLutdd..aananlaylytiticcaall cchheemmisisttrryy.. 22880000..00001100 33MMAA0000223333996655 W WIlLLDDLLI! FF'EE IINNTTEERRNNAATTII OONNAALL,, LLToD --1n1-- PROJECT NO. 454-103 PROJECT NO.: 454-103 Analytical Method AnaTlyhteicmael tMheothdoudsedfortheanalysis of the avian dict samples wasbaseduponmethodology deveTlohpeedmsetthWodiluseedInftoermatthieonaalnaLliyds.isenodf etnhteitlaevdia"nMedtiheot dsaOmutplliense wfoarsthbeaDseedterumpionnatimoneothfodPoFloOgSy developed at Wildlife International Ltd. and entitled "'Method Outline for the Determination of PFOS iinn AAvviiaann FFeeeedd"",. AAvivaiann ddiieettssaammplpelesswweerree eexxttrraacctteeddwwitithhmmetehtahnaonlo,l. MMeetthhaarnoollvwaassaaddddeeddttoo aarereqquiusisiittee qquuaannttiityyooff ffeoeodd ccoonnttaaiinneedd iinn aa FFrreenncchh--ssqquuaarree lgalassss bbooutltlee.. BBootttlleess wweerree ccaappppeedd aanndd sshhaakkeenn oonn a sshhaakkeerr ttaabbllee. `Sampleswerevacuum feredusing qualitative le paper."Thoretainedfoodwasrinsedthreetimes wSiatmhpmleesthwaenroelvnatcouutm he flrs. Thefila wastransferredto filtered using qualitative filter paper. The raevtaoilnuemdetfereidcwflaasskriannseddbtrhoreueghtitmteos with methanol into the filtrate. The filtrate was transferred Co a volumetric flask and brought to volumo vith methanol. As appropriate, samples were futher diluted with methanol. Each sample vthoelunmweaswidtihltmeedthwaintohl.a A50s%apmpertohparnioalte:, s5a0m%plNesANwOepreurfuerthewratdeilrutseodluwtiiothn cmonetthaainnionl.g 0E.1ac0h0 msagm4plHe tPhFenOSwa(stedrinluatledstwainthdaard5)0%anmd e0th.a0n5o%l f:o5rm0i%c NacAidN(Ov/pvu)reso twhaatttehresyofleultliownitchoinnttahineincagli0b.rIa0t0ionmrgan4gHe oPfFtOheS P(iFnOteSrmneatlhsotdaonldoagrdy)./LAamnedt0h.o0d5f%lowfocrhmarict aiscpirdo(vvi/dve)dsiontAhpaptethnedyixfeIll,wFiitghiunrethe| calibration range of the PFOS methodology. A method flowchart is provided in Appendix III, Figure 1. `CCoonncceennttrraattiioonnss ooff PPFFOOSS iinntthhoessttaannddaarrddssaannddssaammpplleesswweerreededteteerrmmiinneeddbbyyrreevveerrsseedd--pphhaasseehhiigghh performance liquid chromatography using a Hewlett-Packard Model 1100 High Performance Liquid pCehrrfoormmtaongcreaplihqu(iHdPcLhCr)omwiattohgaraPpehrykiuns-iEnlgmearHAePwle1t0t-0PLaCckMaardssMSopdeecltr1o1m0e0teHr iegqhuiPpeprefdorwmiatnhcae PLeriqkuinidCEihmreormaTtoorgbroalpohn(SHprPaLyC)iownitshouarcPe.erkiHn-PELlmCesreApaPrIat1i0o0nLsCweMreasascShpiecvteromuseitnegr eaquKiepypsetdonweitBhetaasPielrkCinieanEallmyteircTalurcboollounmSnpr(a1y00iomnmsouxrce2.mmHPILDC,se3papruamtipoanrstiwcelreesiasc)h.ievTedheusiinngstaruKmeenytstpoanreamBetetearssilaCrles asnaulytmicaml caiolnurAmpnipe(nz1d0i0exImd,mTaxbl2e r1.am I.D., 3 lxm particle size), The instrument parameters are summarized in Appendix 11I~ Table 1. CCaalliibbrraattiioonn ssttaannddaarrddss ooff PPFFOOSS pprreeppaarreedd iinn aa 5500%% mmeetthhaannooll : 5500%% NNAANNOOppuurree wwaatteerr ssoolluuttiioonn ccoonnttaaiinniinngg 00..110000 mmgg 44HH PPFFOOSS G(inntteermnaall sstaasnddaarrdd))L/L. aanndd 00..0055%% ffoorrmmiicc aacciidd ((vw/vv)),, rraannggiinngg iinn cocnocnecnetnrtarattiioonnffrroomm 00..00002222991t0o00..00445577mmggaa..i./Lvweerreeaannaallyyzzeeddvwitithhttheesasammplpeless.. TThheessamaeaanmnddmmoeosstt prominent peak response for PFOS was wilized to monitor PFOS in all calibration, quality contro, parnodmstinuednytspaemapklerse.sponNsoeaftotrePmpFtOwSawsamsaudtielitzeodqutaontmiotnyitoPrFPOFSOoSntinhealblacsailsiobrfatiionndi, vqiudaulailtyicsoonmterroilc, acnomdposntuendtys.samLpilneesa. r Nreogreasttseimonpteqwuaatsiomnsadweertoe qgeunaenrtaiftyedPuFsOinSg opneatkhearbeaasirsesopfoninsdeivriadtuioasl (isPoFmOeSric ciontmepmaolnensttsa.ndaLridn)eavrerresgusrestshieonreesqpueacttiiovnes cwoenrceengteranteiroantedrautsioinsg(pPeFaOkSa:reainrteespmaolnsestraantdiaorsd)(PoFfOSthe icnatleimbraaltiosntanstdaanrdda)rdsv,ersuAs tthyepicraelspceacltiibvreaticoonncceunrtvreatioisn prraetsieonste(dPFinOSApp' enindtiexrnaIlI,stFaingduarred)2. ofTthhee cocnacleibnrtartaiotnionstoanfdaPrFdOs.SinAthtyepsicaamlplceaslibwraastidoentecrumrvieneidsbyprseusebsntteidtutiinngAthpepepnedaixkaIrIeI,a Friegsuproens2e.ratTihoes cinotnocetnhetraatpioplnicoafblPeFOliSneainr trheegresassmiponleseqwuaatisond.eterRmeipnreedsebntyatsiuvbestiitounticnhgrtohmeatpoegarkamasreaofrelsopwonasendrahtiiogsh cailnitobrathteionapsptlaicnadbalredslainreeaprrerseegnretsesdioinnApeqpueantdioinx.ITReFpirgeusreensta3taivnedi4o,nrecshpreocmtiavteolgyrams of low and high calibration standards are presented in Appendix III, Figures 3 and 4, respectively. 22880000..00001~1 33MMAA0000223303996665 WWIlLLDDLLII FFEE IINNTTEERRNNAATTII OONNAALL,, LLToD n- - 12- PROJECT NO: 454-103. PROJECT NO.: 454-103 "TThheemmetethhoodd ilinmiittooffquqaunatnittitaattiioonn((LLOOQQ))ffoor tthheesseeaannaallysyesswweaass sseet aattt 11.5 .15ppppmm tthhee pprroodduuccttoofftthhee lloowweesstt ccaalliibbrraattiioonnssttaannddaarrddaannaallyyzzeedd ((00..0000222299mmagLa.i.)/L) aanndd tthhee aa..i. ccaallcuullataetdaeasds ddiilluuttiioonn ffaaccttoorrooff tthhee mmaattrriixx bbllaannkk ssaammpplleess ((550000).). werTeTowwbosoemrmavtaertdriiaxxtbblolraaaanbkkosvaseammtphpleleesLswOweQerrdoeuaraninanalglyyszzeaedmdtptolodeeadtneetaerlrmymisnienesep(poAospsspsieibbnlldeeiixnintIteer,rffTeearreebnnlcceee2ss)...NNAooniiinnmttteeerrffeifrreecnrnceceses inthe feed appeared at approximatelythesameretention timea the peak of ierestbutitwaswell were observed at or above the LOQ during sample analyses (Appendix III, Table 2). An interference in the feed appeared at approximately the same retention time as the peak of interest but it was we!1 bbeelloowwtthhee LLOOQQ.. Figures. AArerperperseesnetnattaitivveecchhrroommatatooggrraammooff aammaatrxixbbllaannkkiiss pprreessceantteeddiinnAApppepennddiixx1II,I, Figure 5. AAvviiaann ssaammpplleess ttoo ddiieett wwaass ddeetteerrmmiinnee ffoorrttiiffiieedd aatt 44..5577,, 118833 tthhee mmeeaann pprroocceedduurraall aanndd 11883300 ppppmm aai..i. rreeccoovveerryy ((AAppppeennddiixx aanndd aannaallyyzzeedd IIIII,, TTaabbllee 33)).. ccoonnccuurrrreennttllyy wwiitthh tthhee SSaammppllee ccoonncceennttrraattiioonnss vweerrsennoottccoorrrreecctteeddffoorrtthhoemmeeaannpprroocceedduurraall rreceovceryooovff9e944.r.77%%y.. mate:fortifiiscpraesteniteodinnAppendixII Figure. AA rreepprreesseennttaattiivveecchhrroommataotoggrraamm ooffaa matrix fortification is presented in Appendix III, Figure 6. AAnn eexxaammppllee ccaallccuullaattiioonn 18.3 ppm a. in avian die. iiss pprreessecntteedd ffoorr .s*aammppllee nnuummbbeerr 445544--110033--22,, nnoommiinnaall ccoonncceennttrraattiioonn ooff 18.3 ppm a.i. in avian diet. Inniitaiall WWeiegighhtt:: 1100.005g FFiinaall VVoolhummeae:: 220000 mmLL DDiilluuttiioonn FFaaccttoo:r: 110000 ((inniteerrmmeedidaiatteedidliuluttiioonn ffacctotrxo ffiinnaall ddiiltuitoionnffaaccttoorr)) PPFROOSSPPaeakkAArreeaa:: 111133556688 IInntteerrnnaallSSttaannddaarrddPPaeakkAAsreea:: 441133116600 PPeeaakk AAvreea RRaatiio:: 00.22778499 CCaalliibbrraattiioonn ccuurrvve eeqquuaattiioonn.. Stop: 277397 ISnltoeprec:ept2:.77030917594 CInutrervceepst:we0i.g0h1t8e9d4(1) Curve is weighted (l/x). PROS ng 41) aor stn PFOS (rag a.i.iL) at instrument PEK 56 00 V-nercp) 15. Conenration = (Peak area ratio - (Y-intercept)) I.S, Concentration TS T ee Slope 22880000..00001122 33MMAA0000223333996677 W WIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,~ LT LTD ne - 13 - PROJECT NO. 454-103 PROJECT NO.: 454-103 _ a7-ooism xor0ms (0.2749 -- 0.01894) 0.100 mg/L 22.7777339977 = ooo main = 0.00923 mg a.i./L Note: LS. =n sandr Note: I.S. = internal standard. } PPFFOOSS((ppppmmaai.i).)iinnssaammppllee. PFOS (ng) at nstrument x Final Volume 1 x Dilution Factor Ce m PFOS (ragv a.i./L) at ine strument x m Final Volum& e (L) x Dile ution Factor Initial Weight (Kg) 0009 x0200100 0.00923 x 0.200 x 100 om 0.01 - 18s ppmai 18.5 ppm a.i. Perntof Nomin. alConti}n Percent ofNominal Concentration = L PEO E GemD) nS se 10 PFOS (ppm a.i.) in sample PFOS (ppm a.i.) nominal 100 is mio 18.5 x 100 = 101% 18.3 During acclimationasdtesting, all HHouosuisinnggaannddEEnvnivriroonnmmeennttaallCConodnidtitiioonnss bids were housedindoors i baer of themosaialy contrDolurlingbroaocdcilinmgapteionnsmanandutfeasctitnugr,edabllybBierdsscownereslhoPursoedducitnsdoCoros (inMobdateterNioe.s Bo735thQe)r.mEosatacthicpaelIny contro!led brooding pens manufactured by Beacon Steel Products Co. (Model No. B735Q). Each pen hEhxaatddefrflnloaoloorwrsasplpaalcc,eetthhalatitnmmgeseaaasnsuudrreefddlaooarppspwpreorrxiemocoaxntesiltyrm7u7c22atXeXtd9e9o0f0lgcacmyml.v.aCneCiizeledidinnsgghtheleiiwgghihttrwwaaanss saahppepeprtoirxnoigm,xaBitemilyac2t233wececlnrm.ye. Esoxrtteerdnalbywawllesi,ghcte,ilintghsenancdhoflsoeonrs iwndeirseccriomnisntartuecltyedfoofmgalwviatnhiiznedcsatceehl wreiprreeasncndtesdheewteinigg.htBciredssswefroer asosrstiegdnmbenyt wtoeigphents,. theEnacchhogsernoupiondfiscbriirmdsinawtaelsy ifdreonmtiflwedithbiyn peeanchnurmebpererseannteddtewsteigcohntcecnltarsastifoonr Iansdsiigvnidmuaelntbto ipweenrrse. ddeEnacishedgrboyuploofbabnidrdss was identified by pen number and test concentration. Individual birds were identified by leg bands. Duringthe estthe averageemperatrinthobroodingcompartmentofthepanswas 3HC +2C (SD).DurAivnegrtahgeeteasmtbthieenarvoeroamgteetmepmepreartautruerefion thheisbrsotouddiyngwacsom2p7a3rtmCen=t o1f2thCe p(eSnDs)wwaitsh3a8Cav*er2agCe e(SlDti).veAhvuemriadgietyaomfbi3en1t r&%oom14te%m(pSeDr)a.tureThfoerpthhoistospteurdiyodw(amsai2n7t.a3inCed+by1.2aCtm(SDc)lowcki)thvaansasviexrtaegeen Hreoluartsiveofhulimghitdiptyerodfa3y1d%uri+ng14a%cei(SmDat).on anThe pthhorotoupgehroiuotd t(hme atienstt.ainTehdebyligahttismuercelocwka)swoarscssicxetenetn hlioguhtrss owfhilicghhtcploesreldyayapdpurroinxgimaactcelimnoatoionn-aayndsuthnlrioguhgt.houtThthee bteisrtd.s Twehreeliegxhptossoeudrcteowaans falvueorraegseceonft lights which closely approximate noon-day sunlight. The birds were exposed to an average of 22880000..00001133 MA0023356 3MA00233968 W WIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTD -14- - 14 - PROJECTNO4.54-103 PROJECT NO.: 454-103 aapppprrooxxiimmaatteellyy 113399 lluuxx ooff iilllluummiinnaattiioonn.. Housing and husbandry practices vere based on gidelins Housing and husbandry practices were based on guidelines eessttaabblliisshheeddbbyytthheeNNataitioonnaall RReesseeaarrcchh CCoouunncciill ((55)).. OObbsseerravtatiioonnss During acclimation al birds were observed daily. Birdsexhibiting abnormal behavior physical During acclimation injurywenortuseed. aFlollbliorwdisnwgetreesotbisneirtvieadtidoanialyn. dBcoirndtsineuxihnibgiutintgiabtneorrmminaaltiboenh,availorboirrdpshywseirceal tinypjuicryallwyeorbesenrovtedusaetd.leasFtotllwoiwceindgailtye.st Ainirtieactoiornd waansd mcaoinnttinaiuninegd oufntailll tmeorrmtailniattyio,ns,iganlsl obfirdtosniwcietrye, atynpdiacbanlloyrmoablserbveeldaviaotrl.east twice daily. A record was maintained of all mortality, signs of toxicity, and abnormal behavior. . mm AniImndailvBiodduaylWboedigyhwtse/FigehedtsCwoenrseummepatisounredatthe initiation ofthe test, on Day 5,Day 8, and on DaysIn1d5ivaindudaDlabyod2y2 fwoerigahltls rwemearienimngeasbuirdesd, aAtvtehreaigneitfiaotidoncoonfsuthmepttiesotn,voanluDesadyur5i,ngDatyhe8e,xpaonsduroen period(Days0-5)andthepos:xposureobservationperiod (Days6-8)were determinedbypen for Days 15 and Day 22 for all remaining birds. Average feed consumption values during the exposure period (Days 0-5) and the post-exposure observation period (Days 6-8) were determined by pen for eachtreatmentgroupandthe controlgroup. Additionally,feed consuwamsdpettermiinoedfnor Deaaychs tr8e.a1t5maenntdgr1o5u:p22afnodrtthheecroenmtaroilnignrgoutpr.eatAmdednittioannadllcyo, nftereodlbcionndssu.mptFieoendwcaos ndsetuermmpintediwoafonsr dDetaeyrsmi8n-e1d5byanmdeas1u5r-2i2ngftohrethcehanregmoiainnitnhge wtreeiagtmhteonft tahnedfeceodntprroelsebnitredds. to tFehedbircdosnsouvmeprtioangiwveans dpeteerrmoiinfetodimdbe.y mTheaesaucrcinugratchyeocfhafneegdecoinnsthuempwtieiognhvtaloufetshmeafyeehdavpreebseeennteadffteocttheedbbyirtdhseouvnaervoaidgaibvleen period of time. The accuracy of feed consumption values may have been affected by the unavoidable wwastaageosoffffeteeeddbabyythtgheebbieirrddss.. GrGorossss NNeeccrrooppssyy Alltestbirdsthatdieddurin thecourseofthetestandallbirdsremainingatthe terminationwere subAjellcttetesotdbi3rgdrsothsastnedcierdopdsuyri.ngAtdhdeitcioounraslelyo,fltihveerteswstearnedwaelligbihreddsarnemdaliinvienrgtaitssthuee,tbelrmooinda,taionndwbiekree wesurbejeccotleldecttoedafgroromsbsirndescreouptshya.nizAeddodintioDnaalyly8,alnivder2s2,waenredwwheeignhpeodssainbdlolfivreormttihssousee,thbaltooddi,edadnudribnilge thweecreoucroslleeocftedthferosmtudbyi.rds euthanized on Day 8 and 22, and when possible from those that died during the course of the study. SStataitsisttiicaall AAnanlalyysseess Moralitydaawereanalyzedusingthecompute programof CE.Stephan (6). Theprogramwas desigMneodrttaolitcyadlaatlaawteetreheanLaClySzOedvaulsuiengatnhedtchome9pu5te%rcpornofgirdaemncoefiCnt.Eer.vSaltebpyhapnro(6b)i.taTnahleypsriosg,rmaomvwinags daveesirgangeed mteothcoaldcuolratethtehebiLnCom5i0alvaplruoebaanbdiltheme9t5h%od c(o7n8fi9d)e.nceTinntethrivsalstbuydyp,rotbheit LaCnaSlDysivsa,lmueovviansg average method detemined using or the thpreobbiitnommetihaoid.proTbhaebilsiltyopemofetthhoed c(o7n,c8e,9n)t.ratIinon-thriesspsotnusdeyc,urthveeaLnCd50resvulatlsuoefwthaes gdetoermoinedodfuifsiintegtteshsetparroobirtempoerthtoedd.. ThBeodsylowpeeiogfhtthdeactoanwceenrtreatcioonm-preasrpeodnsebycuDrvuemaentd'srestuelststuosfinthge TgoOoXdnSeTssATofsoffittwaterset(1a0r,e11)r.epoNrotedst.atisBtiocadlyanwaleyigsehst wdeartea awperpe lctooifmeepedadrceodnsbuympDtiuonnndeattt'as.test using TOXSTAT software (10,11). No statistical analyses were applied to feed consumption data. 22880000..00001144 33MMAA0000223333996699 WILDLIFE WILDLIFE IINNTTEERRNNAATTIIO ONNAALL,, LLTTD0 S15. -15- PROJECT NO: 454-103 PROJECTNO.: 454-103 RREESSUULLTTSS DDiitetAAnnaallyyssiiss AAviavndidiciett sasaammpnplleess wwerereeccoolllecltedefcrfrootmmetthhdee 1188.33 aansdd 11117711ppppmm aa..i. teesstt ccoonncceenntrtartiaotnsiaoanndsd aannaallyyzzeedd ttoo eevvaalluuaattee hhoommooggeenneeiittyyoofftthhee tteesstt ssuubbssttaannccee iinn tthhee aavviiaann ddiieet.t. TThhee aannaallyyssiissoofftthhessee ssaammpplleess aallssoo sseerrvveedd aass vveerirfiicaftiiocnoaoftfittoeessntt ssuubbssttaannccee ccoonncceenntrtraattiioonnss,. RReessuullttiinngg mmeeaann mmeeaassuurreedd ccoonncceenntrtraattiioonnss,, ssttaannddaarrdd ddeevviiaavtiioonnssaannddccooetteicifenftsoioffvcvaraiiraiatetiioonnn(t(CCVVs))ffoorr tthheeseetetesstt ccoonncceennttrraattiioonnss wweerree 11995.5+ +2.21.133 ppppmm aai..i. ((CCVV == 1100.99%%)) aanndd 11119966 + 7700.22 ppppmm aai.i.. ((CCVV == 55.8877%%)),, rreessppeeccttiivveellyy ((AAppppeennddiixx IIl,l, TTaabbllee 44)).. CCoonnttrrooll aavviiaann ddiicett ssaammpplleess ccoolllleecctteedd dduurriinngg tthhee tfesstt sshhoowweedd nnoo iinntteerrffeerreenncceess aabebvetothheevLLOOeQQ. SSaammpplleessccoollleecctteeddudruirninggtthheetteetst t0ovveerriiffyy thhee 3366..66,,7733..22,, 114466,, 229933aanndd558866ppppmmaa..i. est substance concentrations had mean measured concentrations of 402, 74.5, 174, 291 and t5e5s7t pspumbstaan,cerescpoenctcievneltrya.tionThsesheadvamlueeasnrempreeasseunrteedd 1c1o0n,ce1n0t2r,ati1o1n9s, 9o9f.34a0n.2d, 9714..65%, o1f7t4h, en29o1minaanld cc5oo3nn7ccepenpntmtrraatatii.ooi.nn,ss,r,esrrepesesppceeticcvtteiivlvyee.llyy (Appendix TT, Table 5). These values represented (Appendix III, Table 5). 1AA1n0na,ally1ys0si2iss, oo1ff19aa,vvii9aa9nn.3ddiiaetntdssaa9mm1p.p6lle%ess occooflltllheeeccttenedodmffrironoammI ffeeeeddeerrss aaftlterrbbeeningghheelldd aattaammbbiieenntt tteemmppereartauturere ffoorr fiivvee ddaayyss aavveerraaggeedd I1!0311,, 112222,, 110044,, 110011,, 110099,, 111144 and 102% ofthe Da0valyues orthe 18.3,36.6, 73.2, 146, 293, 586and 1171 ppmai. estsubstance concentrations, respectively (Appendix I, and 102% of the Day 0 values for the 18.3, concentrations, respectively (Appendix III, Table 36.6, Table 6). Arepresentative chromatogram of a testsample 73.2, 146, 293, 586 and 1171 ppm a.i. test substance 6). A representative chromatogram of a test sample iisssshhoowwnniinnAApppepndiexIInI,I,FdFiiggiaurroex77.. MMorotalitries atannddCaClilnilniiccaaillOObsbiesrevrcvaattiisoonnss OOnneeiinncciiddetnatallmmorotratlailtityyooccccuurrrreeddiinntthheccaonnrtorollggrroouuppdduuririnnggtthheeccoouurrsseoofftthheessttuuddyy(T(Tabalbele11 aanndd AAppppeennddiixx VV)).. OOnn tthhee mmoornirngoioffnDDagayy 55,, oonnee bbiirdd wwaass nnootiedd wwiitthh a2 bbrrookkeenn lleegg aanndd wwaass ssuubbssecqauueenltlyy eenuthtanhizaedononinDzDaaeyyd66.. AAdddidtiitioonnalalllyy,, ttwwoobbiirrddssiin tthheccoonnttroollggrroouuppwwerereeininteterrmmitittteennttllyynnootteedd wwiitthhffooootk Teleisoionnssaasssociastedwowitithhcccaagigeemmaataetetaagggrgeersessisdioonn.. OOththeerrwwisisee,,aallll ccoonnttrroollbbiirrddsswweereennoorrmmaalliinnaappppeeaarraannccee aannddbbeehhaavviioorrtthhrroouugghhoouutttthheetteesstt.. NNootrtreeaattmmeennttrreellaatteeddmmorotratlailtiitieessoorrovoevretrtssiiggnnssooffttooxxiicciittyywweerreeoobbsseerrvveeddiinntthhee 1188..33,, 3366..66,, oorr 7733..22ppppmmaai.i.. ttrreeaattmmeennttggrroouuppss.. OOnneebbiirdirin htdhee 1188..33ppppmmaa..i.ttrreeaattmmeennttggrroouuppwwaassnnootteeddaasslluammseffrroomm DDaayy 6G tthhrroouugghh DDaayy 88,, aandd bblloooodd ooff udnedteeterrmmiinneedd oorriiggini wwaassnnootteeddoon1~ tthhee uutn~ddeerrssiiddee ooffoovnee bbiirrdd frroomm t5h0e7733..22 ppppmm aai..i. ttreartmeentaggrrootuuppmoonneDDanayyt2222.. OOthter~hviese,raalllwl bbiiirrddsssiinentth,hee 1188..33,, 3366..66aanndd 773.23ppppmm2aa.ii. t{rrecaattmmeenn:t ggrroouuppss wweerree nnoorrmmaall iinn aappppeeaarraannccee aanndd bbeehhaavviioorr tthhroruoguhgohuotttuhhete tfeesstt ppeerriioodd. 22880000..00001155 3MMA0A000223333997700 WILDLIFE INTERNATIONAL, LTD "16-16- PROJECT NO.: 454-103 TThheerreewwaass 1111%% ((11ooff 99))mmorotratlailtityyiinntthhee 114466ppppmmaa.i.i.. ttrreeaattmmeennttggrroouupp,, 8800%% ((88ooff 1100))mmorotratlailtityyiinn the295pm a. amengroup and 100% (10 of 10) moray in the 586and 1171 ps i. the 293 ppm a.i. treatment group and 100% (10 of 10) mortality in the 586 and 1171 ppm a.i. eament groups. Tn te 146 ppm ai. eaten group, ca bid was ulaized on Day 3 afr treatment groups. In the 146 ppm a.i. treatment group, one bird was euthanized on Day 3 after sAusdsiadbnrtohikeeoennvlagesgs.Thil s inciydental, monlywasno sed nthe cautionofthe LCSD abe sustaining a broken leg. This incidental mortality was not used in the calculation of the LCS0 value. one catmenteed moraliinthye 146 pm a. eames group, a bird Additionally, there was one treatment-related mortality in the 146 ppm a.i. treatment group, a bird founddead on temori ofDay7. Clasignsofoxsy wre cerein issen roop found dead on the morning of Day 7. onDay 5, whentwobirds displayed Clinical signs wing droop. Aolf tlooxtihcei~rjbwirerdesaotbstehrivsedteisntcthoisntcreeantmternat twgrieorouenp. on Day 5, when two birds displayed wing droop. All other birds at this test concentration were oma in appasand belo orsdustinof test. normal m appearance and behavior for the duration ofthe test. D1Iaan5t,thhy6eea22n99a9d3p7.pppSmmigaa.n.is.otrfreeaitamreyn.t ggwrreoourupepthftheetrreewcweheerre oeiiggohhntttterhsekamtmmoemnenittn-rgeloaatfteeDddammyoorr4ttaanalidtlie,cs,oooncscirumrirdinntgoobonen cDiatyse5d, 6tharnodug7h. aefxthriboitcend tohfrDouagyh ttShhifeegomnrmsatomohrifnitnsnoiggxnigcoolifeftbyDDiwaadyeyrse8ufrfoivfroirsrvtittonhhgboessueisrninvngeglldleeDoabbnyiirt2ddh2eue.muttbohSraainnngiainnzesgeoddooffonnDDaDayayi4ytya,i8n,danancndcodntltthihnoruuaoueuddggfhhteolttedbhhdeee aapcpfotenpetreaiannrroaaotsnnic,oeon,f, DrrTeeaoddywuueccr8eeddfloirmrrbeetaahccwettiisoeoinnneglttesoo bissarttndiimdmsuuulcliriovniv((isissonoiuguonnunddnstiaalnnDddThaoymmoo2tst2iiino.ognnl))eS,,igrllneeemsttahhoiaanfrriggtynyo,g,xibcwviiitinyrngg.ianpcddplurreodaooreppde,,da0llorossusshffalevoodeff coordination, lower limb weakness and convulsions. The single remaining bird appeared to have reecionvearteidoannd was somalin appearance and bavi from th fem of Day 9 un st recovered and was normal in appearance and behavior from the afternoon of Day 9 until test termination. Jathe 556 ppm a.ramengroupmorales vers frst nk cnDay 3and contined 0be In the 586 ppm a.i. treatment group mortalities were first noted on Day 3 and continued to be cbs trough Day7, at which point all ids hdGd.Over igs of ey wee fs bev observed through Day 7, at wb2ch point all birds had died. Overt signs of toxicity were first observed entheafenaon ofDay 2 and connedhoughthe moming of Day 7, whenthe fal bids were on the afternoon of Day 2 and continued through the morning of Day 7, when the final birds were foJousndofdcedo.Singns dofowieicnyoabscrtvodaomonng b,irdsinthe 586 pr a. resent rpinched 3 rfuofufnldeddaepapde.araSnigcnes,roefdutocxeidcitryoscotbisoenrtveodstaimmuolnig(bsiorudnsdianntdhmeo5ti8o6n)p,plmethaa.ir.gtyr,edaetpmreenstsigorno,upwiinngcldurdoeodp,a ruffled appearance, reduced reaction to stimuli (sound and motion), lethargy, depression, wing droop, lin wesless, ower li rig, prostateposte, and comin. loss of coordination, lower limb weakness, lower limb rigidity, prostrate posture, and convulsions. Jnthe 11po7ma1. reamentgrow 100% mortality had cardbyth mingofDey4. In the 1171 ppm a.i. treatment group 100% mortality had occurred by the morning of Day 4. Signsof toxicity inth 1171 ppm i. traimens group wer iskobservedonthe aro of Dy2, Signs of toxicity in the 1171 ppm a.i. treatment group were first observed on the afternoon of Day 2, viththe fsmorals noted onth morn of Dy 3. Signs offoxy hsv protodes with the first mortalities noted on the morning of Day 3. Signs of texicity observed prior to death includfefid appearerduac nrceaceti,on to simul (sound 30 motion, sy, depression, included a ruffled appearance, reduced reaction to stimuli (sound and motion), lethargy, depression, loss of coordina,win drop, and owes im weskocs adrig loss ofcoordination, wing droop, and lower limb weakness and rigidity. 22880000..00001166 awsoozsasnt 3MA00233971 W WIILLDDLLIIFFIEE II NNTTIEERRNNAATTIIOONNAALL,, LLTTDD 1- - 17- PROJECTNO; s54-108 PROJECTNO.: 454-103 BBooddyyWWeiegighhttaannddFFeeeedd CCoonnssuummppttiioonn `Whencompared othecontrlgroup,therewere noapparent rcamentrete ffectsuponbody weigWhthaemnocnogmbpiadresditon tthheeco1n8.tr3o,l 3g6r.o6 uopr, th7e3re2wpeprme anio.atprpeaartenmet ntrtegartomuepns.t relHateod ewffeecttshvuerpeeonwrabso,ady wcoenicgehnttraamtioonngrebsirpdosnsimvethreedu1c8t.i3o,n 3i6n.6boodry 7w3ei.2ghptpgmaina.io.rtbreoadtymweenitgghrtoulopsss. inHtohewe1v4e6r,, 2th9e3reanwda5s86a pcopnmcenait.ratitorneartemsepnotnsgirvoeupresdudcutrioinnginthbeodeyxpwoseuigrhet pgeariinodor(bDoadyys w0e-i5g)ht(lToasbslein 2thean1d46A,p2p9e3ndainxd V5D8.6 Dipfpfmerean.ci.estrferaotmmtenhtecgornotuopls gdruoruinpgwetrheesteaxtpiostsiucraellpyesriigondifi(cDanatyast0p-5<) 0(.T0a5bfloer2thean1d46Appppmendaii.x lVevIe)l. aDnidffaetrenpc<e0s.0fr1ofmorthtehec2o9nt3roalngdro5u8p6wpeprme staai.tisteivceasll.y sAigsntiaftiicsatnitcaaltlyps<ign0i.0f5icfaonrtt(hpe<01.4061)prpemducat.ii.onleivenl baoncdyawteipg<h0t.0g1aifnocronthtein2u9e3danbde5o8b6seprpvmedaa.ti.tlheeve1ls4.6 pApmstaati.stficsatllcyonsicgennitfircaatnitnt(pr<o0u.0g1h)Draedyucotiofnthine bstouddyy. At weight tghaein2c9o9ntipnpumed atio. becoonbcseenrtvreadtiaont athe s1t4at6isptpicmallay.i.sitegnsitfciocanncten(trpa<ti0o0n1)thrmoeuaghn Dwaeyig8htofltohses csotnutdiyn.ue Athtrtohuegh2D9a3 yppmoftah.ie. sctoundcyenatnradtioanmaarksetadtirsetidcuacltlyionsiignnwiefiicganht g(api<a0w.0a1s) nmoteeadnthwroeiugghht Dlaosys c1o5ontfintuheedstthurdoyu.gDhuDeayot8otoaflthmoertsatluidtyy,abnoddaywmeairgkhetd erfefdeuctcstcioonulindnwoteibgehtdegtaeinrmwiansednoftretdhtehro1u17g1hpDpay a1i5. olfetvheeldsutruidnyg. tDhueeetxoptoostuarlempeorrtiaolidtyo,xbfoodytwheei5gh8t6effaencdts11co7u1ldpnpomt baei.dteterrmeianetdgmfroeornuthptesf1o17r1popsptm- a.i. level during the exposure period or for the 586 and 1171 ppm a.i. treatment groups for post- exepxopossuurreeppeerriioodd.. "Therewerenoapparcatratmentrelatedeffectsuponfodconsumptionat the18.3,36.6,73.2 or 146pTphemre awie.retensotacppaorennt trecatmee(nTntarbetllaete3rd aefanfedcttAspupipeonnodfiexenVdIcsDon.sumHptioon atwthaeer1e8d.uv3c,t3ioe6n.6,ir7n3f.2e,eodr c1o4n6supmppmtioan.wi.atsesntotceodnacenttrhaetio2n9s3,(T5a8b6lae n3d a1n1d71Appppemnadiix. tVrIeIa).tmenHtogwroevueprs, adurriendgutchtieonexpinosfuereed period (Days 0-5). There were notreatmentrelatedcffects on feed consumption in any ofthe consumption was noted at the 293, 586 and 1171 ppm a.i. treatment groups during the exposure period (Days 0-5). There were no treatment-related effects on feed consumption in any of the ssuurrvviivviinngg ttrreeasttmmeenntt ggrroouuppss dduurriinngg tthhee Day Day 6-8 6-8 postxposure post-exposure period. period. In the 293 ppm ai. teatiment In the 293 ppm a.i. treatment grouponlyonebirdsurvivedtoDay22.Thereductionin feedconsumption observeda th 293ppm gair.oupesotncolyncoennetrbaitridosnurdvuirviendgbtootDhatyhe22D.ayThe8-r1e5duacntidon15in-2f2eepd coonsusmpttioenpeorbxisoedrpvsewdaosattsthheeur2e9s3rulptpeomf haa.vi.integsctacoynocneentbriartdioinn dtuhoripnegn,baoth threasDnoatyco8n-s1i5dearneddt1o5b-e22trpeoasttm-eenxtpoesluarteedp.eriods was the result of having only one bird in the pen, and was not considered to be treatment related. GGrorossss NNeeccrrooppssyy DDuurriinngg tthhee ccoouurrsseeoofftthhee tteesst,, aalll bbiirrddss tthhaatt ddiieedd were were subjected subjected to to a a gross gross necropsy. necropsy. Necropsy Necropsy esas forbirdsfounddeadwero similar. Commonobservationsincladd thincondition,lossof rmeussuclltse fmoarssb,irdaslteforeudndspdleeeand cwoleorre, saimutiollayrs.isCoofmtmisosnuesobasnedrvpaatiloensorignacnlsu.dedDetthainilscoofndtihtieon,nelcorsospsoyf fimnudsicnlgesamrassp, ralteersedeinsnpTlaetbelneecd4o.lor, autolysis of tissues and pale organs. Details of the necropsy findings are presented in Table 4. HAofth surviving birds were subjcted to gross necropsy on Day and the remaining birds wereHnaeislroopfstihcedosunrviDvainyg 2b2,irdfsolwloewriengsutbejsetctteedrmtionagtrioossn.nOecnropDsayyo8n, Donayeb8iranddintthehere7m3.a2inipnpgmbirad.s rweeartemneenetrgorposuiepdwaonsnDoatyed2w2i,tfholiaos_lwiginhgtytepsat otelrimvienra.tDionu. (O0tnheDiasyol8at, eodnneatbuirrde oifnthtihsefi7n3d.i2ngp,pimtwaa.is. treatment group was noted with a slightly pale liver. Due to the isolated nature of this finding, it was 22880000..00001177 3MMA0A000223333997722 WILDLIFE INTERNATIONAL, LTD WILDLIFE INTERNATIONAL, LTD se -18- PROJECTNO 454-103 PROJECT NO,: 454-103 notcontsbieredlateedtortreeatmednt. Thesingebirdeuthanizedfomthe293 ppma. reament gnrootucpownasisdeorebdsetrovebdearelnaetcerdotposytreoahtmaevnetw. itThhe sJiancgkleofbmirudsceluethmaansizsedanfrdogmentehreal29t3hinpnpemss.aS.ii. ntrceeattmheesnet fignrodiunpgwscaosrroeblsaetrevdedwiatt hnaecnriopmspyatcothuapvoenwboitdhyaWlaecikghotfpmotuescdlaetmtahsisscaonndcegnetnreartailotnh,intnhessf.indSiinngcsewtheersee fcindoingns costrorebileatterddeawtemitehnrtarneelimadtpeadc.t Nuepcornopbsoydryeswuletigswhterneotuendreamtatrhkiasbcloenfcoernatlraltoioen,rtbhierdfisneduitnghsanwiezreed ocnonDsiadeyr8e.dSitomibleartlrye,atDmaeyn2t 2renleatcerdo.psNyefcirnodpisnygsrewseurletsunwreermearuknarbelmeafrkoaballel fboirrdasll other birds euthanized on Day 8. Similarly, Day 22 necropsy findings were unremarkable for all birds. CCOONNCCLLUUSSIIOONN "TThheeddieiettaarryy LLCCSS00vvaalluueef~oorrnnoorrtthheerrnn bboobbwwhhiittes eexxppoosseedd ttoo PPFFO0Sgwwaassdedteeterrmmiinneeddttoo bbee 222200ppppmmaa..i. wwitithh aa 9955%%cocnofnifiddeenncceeinintteerrvvaallooff 116644ppppmm aa...i. 1t0o 228899 ppppmm a.a.i. TThheeslslooppeeoofftthheecocnocnecnenttrraattiioonnresponsecurvewas 7.008andthechi-vsalquewuaas0r.02e3.The nomartalit concentrationvas r7e3.s2pponpsemcuir.veBwasased7u.p00o5ntarnedattmheenterhei-lsaqtueadrmeorvtaalluietyw, assig0n.s0o9f3.toxTichietynaonmdeofrfteaclittsyucpoonncebnotrdaytiwoeniwgahst g7a3i.2ntpptmhea1.i4.6pBpamsead. upao.nttersetactomnceennttrrealtaitoend,mthoerntoal-ictyb,sesrigvnds-eofffteocxticcointyceanntdraetfifeocntwsausp7o3n2bopdpymwae.ight gain at the 146 ppm a.i.a.i test concentration, the no-observed-effect concentration was 73.2 ppm a.i.. 22880000..00001188 M3MA0A000223333997733 WILDLIFE INTERNATIONAL, LTD WILDLIFE INTERNATIONAL, LTD --1199-- PROJECT NO: 454-103 PROJECT NO.: 454-103 RREEFFEERREENNCCEESS 1. 1. U(UdS.rSa..t)EE,nnOvviPirrPooTnnSmmeeNnnuttamalbl ePPrrroo8tt5ee0cc.tt2ii2oo0nn0:AAggAeevnniccayyn,,Di11s999t96a6.r.y STSoeexrriiieceisst88y55T00e~-stEEccoollooggiiccaall Effects Effects Test Test Guidelines Guidelines (draft), OPPTS Number 850.2200: Avian Dietary Toxicity Test. 2. 2. SUUuS.bS.d.ivEEinsnvivoiinrroonnEmm,eenHntataazllarPPdrroottEeevccattliiuooanntioAAng:geennccyy..Wild11i99j88e22..andPPeesAsttqiiuccaiiddteeicAAssOssreegssassnmmiesenmntst,GGuuisidudebelslioinnceesssi,,onFFII7F1F-RR2AA, Environmental ProtectionAgency,Office of PesticidePrograms. Washington,D.C. Subdivision E, Hazard Evaluation: Wildlife and Aquadc Organisms, subsection Environmental Protection Agency, Office of Pesticide Programs. Washington, D.C. 71-2, 33.. OOTersrgtg.aanniOizzEaatCtiioDonnGuffoiodrrelEEiccnooefnnooormmiTicecstCCioonugopopeferraCathtieioomnincaaalnnsdd. DDGeuevivedeelllooippnmmeee2nn0t5t... P11a99r88s4,4.. Avian Dietary Toxicity Avian Dietary Toxicity Test. OECD Guideline for Testing of Chemicals. Guideline 205. Paris. 44.. AA`SmmueberarciiucctaaenDnieSStooaccriiyeettTyyoxffioocrirtyTTTeeessstttiinnsggwiaatnnhddAvM MiaataneterSirpiaeacllisse..s. 11A99S8877T..M SSStttaaannndddaaarrrdddESPPSrr7aac-ct8tii7cc.ee AffoonrrnuCCaolonndBduoucoctktiinnoggf `SAuSbTacMutSetaDnideatradrsy,TVoolx.ici1t1y.0T4e.stsPhwiliathdeAlpvhiiaan, PA.Species. ASTM Standard E857-87. Annual Book of ASTM Standards, Vol. 11.04. Philadelphia, PA. 55.. NNWaaatstihiooinnnagaltlonRR,eessDee.aaCrr.cchhNaCCtoiouounnncaiclliAl..cad11e9999m66y.. PreGGssuu.iiddee13ff5oorrpp.tthhee CCaarree aanndd UUsseeooffLLaabboorraattoorryy AAnniimmaallss. Washington, D.C. National Academy Press. 125 pp. 66.. SCStoteemppmhhuaannni,,caCCtE.iE.o.n 11997788.. UUS.S.. EEPPAA,, EEnnvviirroonnmmeennttaall Research Research Laboratory, Laboratory, Duluth, Duluth, MN. MN. Personal Personal Communication 7. 7. HSSatteezppahhraadnn,,ECCv.a.EEl.u.at11i99o77n7s7.,.MMeAtmeehthrooiddcssafnfoorrSCCocaailclctucylautfinolrgaaTtnneLsLitCCinSn50gg0.. aPPnadaggeesMsat66e55r--i88al44sI,lnnAAqquPuuaabt.tiicTNTooox.xiiccSoolTlooPggyy6aa3n4n.dd Philadelphia, PA. Hazard Evaluations, Philadelphia, PA. American Society for Testing and Materials. Pub. No. STP 634. 6. Finney, DJ. 1971. StatisticalMethods inBiologicalAssay,20ded.,GriffinPress, London. 8. Finney, D.J. 1971. Statist~calMethods in Bio!ogicalAssay, 2nd ed., Griffin Press, London. 5. Thompson, W.R. 1947. BacieriologcalReviews, Vol 2, No2: 115-145. 9. Thompson, W.R. 1947. Bacter~otoglcalReviews, Vol 2, No.2:115-145. 1100..wDDiuutsnhnnaeetCt,o, nCCt.rWWo.l. Jo11u9r95.555.A.meArA. MMSutluatltsii.pplleeAsCCsooocmm.pap5ra0ir:si1soo0n9n'6'ss-1PP1rr2oo1cceedduurree for for Comparing Comparing Several Several Treatments Treatments with a Control. Jour. Amer. Statis. Asso. 50:1096-1121. 11. Gulley, D.D. 1990, TOXSTATRese 3.2. The UniveofrWsyoimitngy. 11. Gulley, !).I). 1990. TOXSTAT Release 3.2. The University of Wyoming. 22880000..0000~199 33MMAA0000223303997744 W WIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDD 20- - 20 - PROJECT NO.: 454-103 PROJECT NO.: 454-103 TTAABBLLE E|1 CCuummautlaitivvee MMorotratalliittyyffiroomm a a NNoorrtthheerrnBBoobbwwhihitteeAAccuuttee DDiiectaarryy TTooxxiicciittyy Sudy Study with with PFOS PFOS --Fp-- anen-- al Group Experimental TNoo.DEDxeepaaoddsPPueerrreNNPoeor.EiExoxdppoosseedd NNooP.oDDseteaEadxdpPPoeesrruYe Nreoo. PEEexxrpipe ooodsed Group Exposure Period Post-Exposure Period - emai) (ppm a.i.) BDa5y 00 DDaHyT1 DDany72 DDany 33 DDawyi4 Dus Day 5 DuDaey 6 Dl Day 7 DDaayv8 t -- Contra Control 0 0 00/30 030/30 O00/300 O0/3300 0%0/30 0%0/30 101/30 11/8300 101/30 Treatment Treatment 183 wo ono ono G0 ol ol ol 010 0M0 18.3 366 wo om ow 010 oo oo 0 0 00 36.6 2 ol ol oo ono 00 ol 00 010 010 73.2 146 oo ono ono we oo 09 w ww 146 2 on wo ono ow om 20 40 810 80 293 8 op om ow M0 20 so so 100 1080 586 wm oo ono 00 0 M00 wi 00 1010 1010 1171 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/I0 0110 0/10 0/10 0110 0/10 0f10 1/102 0!10 1/10 3110 0/10 0/10 0!10 0/9 0/10 2/10 10/10 0/10 0110 0/10 0/9 2!10 5/10 10/10 0/10 0110 0/10 0/9 4/10 8/10 10/10 0/10 0110 0/10 119 8/10 10/10 10/10 0/10 0/10 0/10 1/9 8J10 10/10 1~/10 TAve ied BET5018 357Gin: eraG10 po a1 02007e m 1 ee The LC50 value was calculated to be 220 ppm a.i. with a 95% confidence interval of 164 ppm a.i. to 289 t,pm a.i.. Tonle ecard aothe chlo eset gros aDay 0Dy22. ~ - No mortalities occurred in any ofthe control or treatment groups from Day 8 to Day 22. Bidcanis onday 3sles staining sben cg 2 _ Bird euth~nized on day 3 aRer sustaining a broken leg. 22880000..00002200 3MMA0A000223333997755 W WIlLLDDLLI! FFEE IINNTTEERRNNAATTIIOONNAALL,, LL-nD::) 21-21 - PROJECT NO.: 454-103 PROJECT NO.: 454-103 TTAABBLLEE22 PPaaggee!1 MMeeaannBBooddyyWWeieigghhtt ((gs)) ffrroomm aa NNoorrtthheerrnn BBoobbwwhhiittee AAccuuttee DDiieettaarryy TToorxiicciittyy SSttuuddyy vwiitthh PPFEOOSS Bem Experiment~l Group Group Goma) (ppm a.i.) EExxppoossuurree PPeerriioCoddhangs Dio Day 0 Dus Day 5 Day0S Changet Day 0-5 PoPosstt-EExxppoossuurrCsehPPaenergie?odd DDanys8 Duss Change~ Day 5-8 Toul Total Clam Change1 Contra Control 0 Men 0 ww s 1 0 Mean 20 30 10 38 8 18 1s 3 5 2 + SD 1 4 3 5 2 4 Treament Treatment 1188.33 EMMyaeann a211ao3m1 1u1 3 405 29 220 3 SD 1 4 3 5 2 3 ss36.6 EMMYeeann 220EREa31 u11n2 393 58 1 1129 SD 2 3 2 3 1 2 B273.2 EMMYeeawn 02012w3n0 99 1 7373 77 1 11266 SD 1 2 1 3 1 2 6 Man 0 mp 3 ee 1 146 Mean 3 2s 3 3 2 SD 20 27* 7* 2 3 3 33* 6** 3 2 13"* 4 2 Mem 0 ae ae ean an 293 Mean 12 2 a + 5 SD 20 18"* -2** 1 2 2 18"* 4 -2** 4 -1"* 5 $6 Mean 0 ae an - - . 586 Mean EY 2 2 : : : SD 20 16"* -4** 1 2 2 wn ven 0 - - . . 1171 Mean 2O -- Ey 1 : : : : : SD 1 Vien hen Fs alcutod spayTos 5 se odywelts wg AVRO Soywes (5 Append VD. 1Mean change is calculated separately from the mean body weights using individual body weights (See Appendix VI). )=Nodtsvidoe tomrt (-) = No data available due to mortality. Staal diff em he contolgop a: p<0.05 Dunne' est). *Statistically different from the control group at p < 0.05 (Durmett's t-test). Sinisa fre oe th oil ropa p00](Dues ist). **Statistically different from the control group at p < 0.01 (Dunnett's t-test). 22880000..00002211 3MMA0A000223333997766 WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LTD LTD -n- - 22 - PROJECTNO. 454-103 PROJECT NO.: 454-103 TTAAPaBBgLLeEE222 Page 2 MMeeaann BBooddyy WWeeiigghhtt (2g)) ffroomm aa NNoorrtthheerrnnBBoobbwwhihtitce AAccuutteeDDieiettaarryy TTooxxiicciittyy SSttuuddyy wwiitthh PPFFOOSS Emme Experimental Group Group Gpmai) (,ppm a.i.) CCoonnt0trrooll 0 Treatment Treatment #318.3 3636.6 B273.2 us146 3293 FMME eeamn SD Mem Mean SD BM M)eeamn SD M MB)eeamn SD Mem Mean 0SD M p5eeann --PPoCe-- sht-maEnxwgp?oesuererPeeriG odd a args Toul Days Day1s Daysds Changet Day 8-15 Duyn Day 22 Deyis: Change~ Day 15-22 Change(22 Total Change1(8-22) Day 8 Day 15 w37` o5190 2o3z5 6 10 5 2 as 82 22 45 5 3 8 13 3 8 040` 6o8e8 2o4w3 6 8 3 3s83 oe655 226 3 5 2 3s 53 w604 2ou42 3 4 2 3w43*oos583 o2u41 3 3 1 221 335 : u14 a 2 a 87 23 47 7 3 3 7 3 3 89 7 2x42 55000 7 2 4 7E94l 202 4"4 2 4 2 2 7"92 221 2 a451 2 2 1 B5l5z 20 EF 34 SD 586 Mean : : . : : SD um Men - - - . - . 1171 Mean - - e E) i : e : : - re: e SD - - - iSha eed SyTo hm oly eign vn Oyweg (5s Append VD) tMean change is calculated separately from the mean body weights using individual body weights (See Appendix VI). ne cold notbe calsiedsaiscally ihDunes 5. 2n=l. could not be evaluated statistically with Dunnett's t-test. =Nodata ealde to matty, (-) = No data available due to mortality. "Saistaly dillcnt ftohe cmone prop at p< 005 Duna' es) *Statistically different fi:om the control group at p < 0.05 )(Dunnett's t-test). 22880000..00002222 33MMAA0000223303997777 | WILDLIFE INTERNATIONAL, Lo WILDLIFE INTERNATIONAL, LTD a. - 23 - PROJECTNO. 456-103 PROJECT NO.: 454-103 TABLE TABLE 3 PPaaggee 1| Mean Feed Consumption ghia) rm a Noster Bbwhic Ace icary Mean Feed Consumption (g/bird/day) from a Northem Bobwhite Acute Dietary `TTooxxiicciittyy SSttuuddyy wwiitthh PPFFOOSS emer Experimental GGrroouupp ((ppppmm aa.ii.)) Ey Control .0 Ee EExxppoossuurree PPeerriioodd Day 0-5 j5o s: Mean 9 SD 2 Tepe PPosotst]-Exposure PPeerriioodd Day 6-8 0)10 2 -- Treatment 13 , W 18.3 9 11 366 ' u 36.6 8 12 na 0 i 73.2 10 13 wo s 0 146 9 10 s , 293 5 9 0 ` 1 586 6 19 wm ` . 1171 4 TE (-) = No data available due to mortality. 22880000..00002233 awsoozsas7s 3MA00233978 WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIO ONNAALL,, LLTTDD _ 24- - 24 - _ PROJECTNO.: 454-103 PROJECT NO.: 454-103 TTAAPaBBgLLeEE?S3 Page 2 MMeeaann FFeecedd CCoonnssuummppttiioonn ((Tggo/bnbiiirrcdid/tddyaaySy)t)uffdrroyommwiaathNNoPorrOtthhSeerrn BBoowbwhihtitse AAccuuttee DDiieettaarryy Toxicity Study with PFOS Experimental Group Group pms) (ppm a.i.) -- Control 0 MMEeaeann SD ow PPoosstt-EExe xppoossuurree PPeerriioowdd nwm Day 8-15 Day 15-22 929 1131 2 1 T~catment 18318.3 16636.6 273.2 6146 ES293 ss586 mm1171 10 2 10 12 1 15 14 15 5 15 n13 u15 11 14 8 9 8 - - - - O Nodamedem (-) = No data available due to mortality. 22880000..00002244 3MMA0A000223333997799 W WIILLDDLLIIFFEE INTERNATIONAL, INTERNATIONAL, LD LTD 2- 255.- PROJECTNO. 454-103 PROJECT NO.: 454-103 Group GroTTsAsABPBaLLthEEo4l4ogical Observations ffrroomm aaNNorotrthheerrGnnrBoBouobpbwwGhhiriottseesAAPccauutthteeolDDoigiceittcaaarrlyyOTToboxsxieicrcviitatyytioSSnttusuddyy wwiitthh PFOS PFOS Died tha ieddurinhge uresofthe sy e me rVile,e Female, aud Undecrmited Birds that died during the course ofttae study Male, FemalPe,RaMnAd LUndetermined FFiinnddiinngg CNoe l Control B N1W4ee6 N29=3PPMW A.I.N5e8l6o TN1eW17l1o N=I N =2 N =8 N =10 N =10 AAAbbbdddooommmiininnaaalll avy, some ccaavviittyy,, asoomle aauuttootllhyysrsioissughout o 0 o o 0 0 2 02 2 2 1 ` 4 1 1 Abdominal cavity, autolysis throughout ECmroocpicsaepdty Crop, empty 00 0 o0 20 2 0 0 2 s1 5 5 2 52 0 1 0 1 2 0 05 08 EFFGrrmLaacactcwuuiararceetetdddellmeopggty GGi.Iz.atrnadcctoenmtepnttys bile stained o1 0 0 o1 00 0 1 21 0 1 51 0 0 0 1 Gizzard contents bile stained HeHeaarr,,panotleeriorportion matted whitecoor Heart, anterior portion mottled white cotor 0 0 0 o HIneeasrti,npaallceontents ack 0 o 0 0 0 0 20t1 5 o o2 00 I 1 0 o o0 2 2 o1 IKnetse)s,tipnraolmcionnetnetnts tar-like KiKKLdieindveenel,ye,pypsrs,lo,pempaaalilnneeedntmote o0 00 0 0 0 0 0 1 o1 2 5 3 2 0 1o10 0 0 0 1 00 2 0 o0 Liver, pale and mottled Loss of muscle mass Loss of muscle mass Muscular skeletal, pale Muscular-skeletal, pale 0 o 00 o1 0 1 0 4 4 o .0 707 0 9 09 0 0 01 0 3 0 0 0 0 Small in stature SSpmlaclnl,inblsatactkure SSppllecean,, bdalarckk 0 0 0 o 0 o 0 0 03 00 0 1 o1 0 0 0 2 Splecn, Spleel~, Spleen. Splca, dgraerky ggrreeyy-brown 0 o00 " 0 0 0 0 0 o 0 o 0 1 o1 02 0 1 0 o 0 0 0 1 0 o 1 1 SSSSpppplllleeeeeeennnn,,,,pgpsamralealeeyl-lbrown SSpplleeeenn,, ssmmaallll and pale 0 0 00 0 0 0 o o1 0 0 00 0 3 1 1 01 Spleen, small and pale TNhoit nRemarable Thin 000 " 00o0 000' 34`o 2o020 Not Remarkable 0 0 1 0 22880000..00002255 33MMAA0000223333996800 = 3 i 58588 58 88 838 2% 3% 5% 5y 8 2i gE EER $58813% 5 85 55 2% % 55% 8 i , [715 sussss 55 88 53 83 28 53 8 ; 3 i o3f *gg eg SggE BgEg g80g EE 1i i: 585888 285 85 5 5% 2% 8% EH"yy Ei 588885 $8 83 8% 58 5% 8% 885 3 [Fi fiz 2 3ig gregsRiiicicdl | E3 3 88558 2 88 88 8% 8% 2% R33 Bl ii sssgss ss $3 eggs 3% 5% se 5% 88% 938835);81i .74 zs fl :Zz co 32 88 38 82 38 29% 1 e i 2 || ssssss 5 85 85 88 28 332 Ll = qi 5 | 555388 33 55 83 535 5% 8% sefs1115 23 Z H2 45 ~emeee mn mma ma ma ma me 13i 13 gZ z i1E:Ed3E: cocccoyiz=z 982 R8F Hii E8883EEl E[Zl i EY Z i5 4 FEE 3 z8J 22880000..00002266 awaoozsaset 3MA00233981 WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDO -2217-- PPRROOJJEECCTTNNOO..:: 445544--110033 = APPENDIIX APPENDIX I CCeerrttiifficattee OOff AAnnaallyyssiiss FC1~-C9-595,, LLoott 221177 DE AST Resinsre M Mara~hr99,~c2200h 0000 Em ee-- s ------ Anioneore notnETgveeii,PRib e Nr m[m | o"ea| [emi CER, Er Cm 22880000..00002277 amaoozsasez 3MA00233982 W WIlLLDDLLII FFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDD --228 -- PPRROOJJEECCTTNNOO..:: 445544--110033 AAPPPPEENNDDIIXX IIIf WILDLIFE INTERDDNIIAEETTTIFFOOONRRAMLMUULLTLDAA,TTIGIOOANNME BIRD RATION WILDLIFE INTERNATIONAL LTD. GAME BIRD RATION1 IINNGGRREEDDIIEENNTTSS SaFFiiynneeBCCeaoonmmMMMeeaealal,l48% Protein PrWW SohotheyeeaBaittenMaBMniaiddsMddesesal, 48% Protein APAArglgowtlaeaiayyn MSBSppaeeesc,ceiiaal2l,,066%00%%PrPPortrooictneiinn ADGDrrlrifoieaeuddlnfaWWdhMLheieeymyael,s 20% tone Protein GEMEeaartsoshttumimnoaadnnniLnCCiemaalPlePPsrhhtoooenssme + Liquid VMGViLi~ttataFhmmiiooinnrmtiaannn(eddFPMeMrimenimacnlreixearals+l)PPLrrieeqmmuiiidxx ((ssecebbeellooww)) GL Ferm (Fermateo)2 TSoalutllodized Salt Iodized Total PPEERRCCEENNTT ((%%)) 4340.688533 30666..05650050 4346..00000000 0232..55050000 0000..36965000 0000..23335252 100000..10210500 100.00 V`VITITAAMMIINN AANNDD MMIINNEERRAALL PREMIX PREMIX AMOUNTADDED PERTON AMOUNT ADDED PER TON VVViiitttaaammmiiinnn ADDy3 RNiVRibaiiobtcafoimlfnlaianvviAinn NPVPaaiinanatctnooitnithhaeennBiiicc; AAcciidd BiVFFoooilttliaiiccmnAAincciiBdd12 PTyBPhryiiiordaitodimnxoiixnnignee TVVVihaitiaanammmiiiiannneKEE (Menadions Vitamin K (Menadione ZMianngcanese Manganese lCZCooiopnnppc peeerr IIISooreodlnnienneium. Selenium Dimethylpyrimidinol Dimethylpyrimidinol Bisulfite) Bisulfite) 202,000000,00,000000000 LLI.UCC...U. 7,000,000 4606 ggLgrrrUaaa.mmmsss 4110800 mgggrrreaaasmmmsss 660064004812mmmrmageegggrsssssams 20,001110..2221g.ggrrraaammmsss 20,01000255.88Ig.rUggar.rmaasmmss 144077268gggrrrgaaarmmmasssms 51611..585grgggarrrmaaasmmmsss 11558122 mgsgrams mgs 1"TThhoeegguf9ua%arracanrntuteedeeeddiaeannra.allyyssiiss iis aammiininmuimomoff2u217m%% pprrootteeiinn,, aammiinniimuummooff 22.5%.c5%rcurudde fefaatt aannddaa mmaaxxiimmuumm: 2*FFeeomrfme5ne%tnatatctirioounndeBByfyi-b-PPerrro.odduuccttss ((SSoourucreooffUUrindidenetnitiffiieodd GGrroowwtthh FFaaccttoorsrs).). 22880000..00002288 33MMAA0000223333996833 WWIlLLDDLLII FFEE IINNTTEERRNNAATTIIOONNAALL,, LL'TIDD 9- - 29 - PPRROOJJEECCTTNNOO..:: 445544--110033 AAPPPPEENNDDIIXX IIIII ANALYTICAL METHODS AND RESULTS ANALYTICAL METHODS AND RESULTS 22880000..00002299 M3MA0A002023333996844 WILDLIFE INTERNATIONAL, Lo WILDLIFE INTERNATIONAL, LTD -30- 30 - PPRROOJJEECCTT NNOO.:.: 445544--110033 - INSTRUMENT, AANNALAYLYTITICCAALLCCOOLLUUMMNN:: OVEN TEMPERATURE: O~NTEMPE~TURE: STOP TIME: STOP TIME: FLOWRATE: FLOW RATE: MOBILE PHASE: MOBILE PHASE: IINNJJEECCTTIIOONN VVOOLLUUMMEE:: PFOS RETENTION TIME: PFOS RETENTION TIME: IRINENTTTEEERNRNTNAIAOLLNSSTTTIAAMNNEDD:AARRDD RETENTION TIME: PFOS MONITORED MASS: PFOS MONITORED MASS: IIMNNOTTNEEIRRTNNOAARLLESDSTTMAAANNSDDSA:ARRDD. MONITORED MASS: AAPPPPEENNDDIIXX IIIllI TTaabbllee 1 TTyyppiiccaall LLCC//MMSSOOpepreartaitioonnaallPPaarraammeteteerrss HHaePewewirleektnti-PPnaac-ckEkaalrrmdAdePM MrIoodd1e0sl0L11C1100M00aHHsisiggShhpPPeceetrfrroorfmmeotaernrcmeeqLaLuiniiqqpcuupiieedddwCCihhtrrhoommaaaiPtoeorggkrriaanpp-hhElwwmiiethrh T(aTSuuPIrreMhbr)ook.lIioonnn-ESSplpmeraaeyyr fon source. API 100LC ion source. Operated in selective ion monitoring mode Mass Spectrometer equipped with a Perkin-Elmer Operated in selective ion monitoring mode KeKyesytstoonneeBBeteatsaislil CCy~s8ccoolluunmma ((110000 mmumm xx 22mmamn IL.DD.,,33 ugmanppaasrtitcleessiizzee)) 3E0XC 10.0 minutes 10.0 minutes 0220 minute 0.220 mL/minute 7722..00%%MMetehthaannoo!l :: 2288.00%% NNAANNOOppuurreeWWataeterrccoontnataiinniinngg00..11%% FFoorrmmiiccAAcciidd 2255.00~4L Approximately 7.0 misutes Approximately 7.0 minutes AApppprroosxiimmaatteellyy 44..8 mmiinnuutteess 449988..66 aaminuu. 267 amu 426.7 ainu 22880000..00003300 33MMAA0000223393998855. r WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LL-Io"D 1s -31- PPRROOJJEECCTT NNOO..:: 445544--110033 AAPPPPEENNDDIIXX IIlI Tabie2 Table 2 `Mtr Matrix BBllaannkkss AAnnaallyyzzeedd CCoonnccuurrrreennttllyyDDuurriinnggSSaammpl~llee Asalysis Analysis J 5SN4au2nm1tb0ee5rr) SSaammppilee Tipe MTieeassurreeddGpPCCoFrooOmonnsSaca!~e)rntaraitioonn ooff (ppm a.i.) MAB-1 (454-103-) MatixBank Type Matrix Blank << 1LoOQQ MAB- 1 MAMABB2-2 Tt of quan(C00)wes M Maattirixx BBllaannkc 1p1p 5a3 Ease upon procs of the <10Q os<tLOcaQlfbration Sandard any 00072 mL) ae - ~ The limit of quantitation (LOQ) nnzlyzed (0.00229 mg a.i./L) and he dilion ctor was 1.15 ppm a.i. the dih,tion factor of themst banksamples GOD) based upon the product of the lowest of the matrix blank samples (500). calibration standard 22880000..00003311 M3MA0A002023333996866 WWIiLLDDLLII FFEE IINNTTEERRNNAATTIIOONNAALL,, LTD LTD ne - 32 - PROJECT NO. 454-103 PROJECT NO.: 454-I03 APPENDIX IT APPENDIX III Ties Table 3 e MaMtartirixx FFoorrttiiffiiccaattiioe onnssAAnanlaylyzzeeddCConocnucrurrernetnltlyy DusuirinnggSSaammpn plleeAAnnaalyyssiss Sample Number ppm Percent. Sample Number @stei03) Forel Nica Recovered (454-103-) y wen aw we MAS-1A MASA i in 105 MAS-4A Concentrations of PFOS (ppm a.i.) Fortified Measured 4.57 4.54 4.57 4.79 Percent Recovered 99.2 105 Masa 183 16 251 MAS-2 183 176 96.1 MASS 15 EH MAS-5 183 162 88.3 MMaAAsSSs-S3 111585533000 171155177666 268617.1 --MAS-6 1830 TT Sunda1r7d1D6eviM m ateiaonn -= e99934..77 Standard Deviation = oNv=e CV = 6.99 `n7.38 "Nootte::RR0eldsmusltsaa5nlddcccloorrcertecciitoioonnnsssToforVraneheewws tohealstvesbbuebssotaarnnoceuPnpduuerrditfyyoWweererpeerGtgoeinnnegcriapteeuddpGuassni:ngNgN=MAaEcQQuuaann veron6 version 13 1.5 savare software and manual calculations. Values have been rounded for reporting purposes. 22880000..00003322 M3MA0A002023393998877 g 1 i : ol 5 gi g . t z Be |i oe | 3gf185 = i88 |i 8 i i fi EEE] 5 i g I: : 1 Ja Resdl i 1AEE338882EREaFuEEg!EY !i g #23 HH b&Z fie H+iie = i1:E 33b.I $3 22880000..00003333 -- 3MA00233988 z : : iy g 3 les oss oot 11 2 is ; : Hi3 oeo sensi ii i3 i Bl : E 1 dill i pepe. 1 il ween] : tl $i i i i3 B|f cocE oeog eel ec |e b g | Ii J : Lh 15 ~ o~ RL i Bilees = = 5 sfifdl Ld 3 I 14 s J 2800.0034 -- 3MA00233989 . 2 z i pillrre eee i g i 2 HTdsgigLLe.rred { I i3 3 i3 f3i8 fea2 3= 225223 99 88) iH i i z sil 3 3 Hie on ove zn mm on ol i Tei Hl i i He: 282g 38% i ! [322 ; Plz ere eee " 3 Z BE z EByl-.v orz ioz s2 e oz og i iif z 1iz 228 23 22880000..00003355 M3MA0A000223333999900 W WIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDD 36 - 36 - PPRROOJJEECCTTNNOO:.: 445544--110033 APPENDIX III METHOD METHOD OOUUTTLLIININNEE FAFOVORIRATTNHHDEEIAAENTNAALLYYSSIISS OF OF PFOS PFOS IN AVIAN DH~T Peps ition pls he sed vindstockwin ey mi edie Prepare matrix fortification samples in the desired avian feed stock using the dry mix technique. yivTies Dry Mix Technique rihiha le n WIA n rr1 a (is, ih For the high-level matrix fortification sample, weigh the requisite qumtity of Wildlife International Ltd. (WIL) ration into a weigh boat. pitty es108 se Weigh the requisite quantity oftest substance (PFOS) into a beaker. lt Wn ct re be Add ofthe WIL ration and the te~t substance to a larger beaker. mtbr int FS wit cn foci rs dies rrbe - Rinse the beaker that contained the PFOS with small portions ofthe remaining ration and ~rmasf'er all portions to the larger beaker. bnct il o te essr Wig Mix the contents ofthe larger beaker well and tr0msfer the mixRtre to a Waxing blender. Bio inn 3 ls seg 1 ioe cr scedovesofde ede Blend the mixlure for ~5 minutes stopping at 1 minute intervalz to scrape dovm the zides ofthe Mender. Duin hid il rt ed oboe,pa= rhreSeebscons in fb During the third interval transfer the fort~ed feed to a beaker, mix well and return the mixture to the blender to complete mixing in the specified time. cpa es tiebr ys Bs ic Prepare the next two matrix fortlfieation levels by eeria| dilutions. Fontesect sad gh oe ms caocr ehopeteulyof ridmixigh Follow the same procedure described for the high-level matrix fortification except weigh the appropriate quantity of fortified matrix (kigh ~r ide fr mid-level) raffler than the test substan,. Weigh 10gspe thsish, sic voito sRdemsepineel.ih bossad si 6.rcha Weigh 10-g samples ofthe matrix blank, matrix fortification and test samples into weigh boats and transfer to 16-oz. French-square glass bottles. Record the weights. o mrs0ii f oe df fr et Fe For each sample, measure 100 mL~ ofmethanol with a graduated cylinder and transfer volume to the French-square bottle. Cp dp t ir. rd ie 0s 70 Cap bottles and place on shaker ~able. Allow.the samples to shake for a minimum of 30 minutes at 250 rpm. Vil choc i gli ho ie od 3i ml hh Vacumn filter each sample with qualitative filter paper and rinse retained feed 3 time~ with methanol Inlo tho filtrate. Tic let.20 ml.voli lk sin sk eli ihml. Tramer the filtrate to a 200-mL volumetric flask and bring the flask to volume with methanol. Fp spc it)fo bin sort` cess of LEMS oils. Prepare appropriate dilution(s) to bring final con~ntration into the calibration range ofthe LCMS methodology. me eitds ees Use methanol for intermediate dilutions, ifrequired. Frfli 00hS08NANO wrsoln sin 100 mg 4 FOS(trlsit). I~, ~ It fi,~M ct;h,t;t~,, ,,~ ~;oo/~ m~lhannl : gOo/~ NlAMOnur~ water solution conlalnlncr O. 100 m~ 4H PFOS (internal standard3/L Tom formed and 0.05% formic acid (v/v). Amp sd bestsrsorLOMSslic. Ampulate and submit samples for LC/MS analysis. FFiigguurre e11.. AAnanlayltyiticcaallmmetethhooddfflloowwcchhaarrtt ffoorrththeeaannaallyyssiiss ofPROSinavian of PFOS in avian dit. diet. 22880000..00003366 33MMAA0000223333999911 WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDD 3- 3774APPENDIX TIL APPENDIX III PROJECT NO: 454-103 PROJECT NO.: 454-103 11.5500 o 1001.00 . 050 0.50 000 Herre 0000 0100 0200 0300 0400 0500 0.00 ......... 0.000 0.100 0.200 0.300 0.400 0.500 CCoonncceenntrtraattiioonn ((RRaattico)) FFiigguurree22.. CAAutrtyyvppieiccaiasllcwceaialgibhltraeitdio(bn1cc)urur.vraveeftfoorriPPEoFOOSnS.. SSllooppee = = 277397;Itcrcept 2.77397; Intercept = = 0.01894; 0.01894; 1 r = = 0.9981. 0.9981. Curve is weighted (l/x). 22880000..00003377 33MMAA0000223333999922 WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDO --3388-APPENDIIX APPENDIX III PPRROOJJEECCTTNNOO..: 445544--110033 10lO0o9O58O" 77(7 o60~ 550. 4~ 5 220" 10" a2 Tho01 1 .~0 inonchy: 1995 cps 187 intensity: 1995 cps 67 us145 73s61 2.55 = 122 .= = ___,.,. ..... Shr91 3.81 Gs 121 5.06 Hsrm\, ..~ 151 6.32 w7es aas ~,,, :~ _-~'__.__-~--: -~_ 161 7.57 2";1 8.83 sT~ aen Time FFiigguurreed3.. AA rreepprreesseennttaattiivvee fioonncchhrroommaattooggrraammooffaa llooww--lleevveell ((00..0000222299 mmgg a.) a.i./L) PROS stand. PFOS standard. 22880000..00003388 3MMA0A000223333998933 WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDD 39- - 39 - APPENDIIXL APPENDIX III PPRROOJJEECCTTNNOO..:: 445544--110033 10 166 iinnttoonnssiityy:;2266995544cps 66 100" a90- 580- 77O" o 55O" 40" 53O" 22~ + o 0 ," 24 as 75 ~, , TEL er 81 0 2s 3m 1.30 ' 6'1 ' 2.55 ' 9'1 ' 3.81 144 144 2 ,~._~, aE aL 121 Gs 6% 5.06 , ~ 151 6.32 190208 __ 230 ~.19,02,04, .230. isi eli | Sm 181 7s ss Tee 7.57 211 8.83 Scan Time FFiigguurreed4.. AArreepprreesseennttaattiivveeiioonncchhrroommaattooggrraammooffaa hhiigghh-lveevlel((00..00445577mmgg 2a./i.1/L))PPRFOOSSstsatannddaarrdd.. 22880000..00003399 M3MA0A002023333998944 WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LTD LTD 40- - 40 - AAPPPPEENNDDIXIIIXITI PPRROOJJEECCTTNNOO:.: 445544--110033 100 sol a 24 s0 . "1w8o0 86 nintteonnssiyty::223300ccppss 50 50 0 #4 144 144 20 216 E 1LA1 Ady AS MAALpt E Le 2h STU i^^ f ~T'e'lF V " VTen" tl us 1N~_~kl igiII t..t~t~ w.i I,#~Vaiwr v" \It gn 3T1o S6h1 9S1 m See 121 5.06 ee151 6.32 Te181 7.57 es 211 8.83 Tm~ Time 1 2.55 3.81 FFiigguurerSeS.. A"ATrehreperpaerresrseeonntwatiatntidivivceeaccthhersrootmmhaeatrtootggrrtaaommonofftaaimmmaetaortrfiiPxx ObblaSarnkk sample(454-103-MAB-D). sample (454-103-MAB-I). The arrow indicates the retention time of PFOS. 22880000..00004400 M3MA0A002023333998955 W WIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTDD a-4n1eAPPENDITXL APPENDIX HI PPRROOJIEECCTTNNOO:.: 445544--110033 To 168 Inintetennsistityy::66223388ccopss 166 100" 59O" 88O" 77Oa 5 rt 4~ 3 144 2 144 1 2244 112222 S311%1616218 99a11s 11s32o11s 1165511% 117885117 228211111s3 | TSScmeamen 1.30 2.55 3.81 5.06 6.32 7,57 8.83 Time FFiigguurrees6.. AArerperperseesnetnatattiivvee cchhrroommaattooggrraammooff a mmatartriixx ffoorrtiiffiiccaattioonnssaammppllee (454-103-MAS-14) (454-103-MAS-1A). 22880000..00004411 33MMAA0000223393999966. W WIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTDD a- - 42 AppENDDCI APPENDIX III PPRROOJJEECCTTNNOO..:: 445544--110033 To 167 InInttoennsshityy:: 7248cps 7248 cps lOO- 67 99O- o80- 27o- 6of 0- 550- 44(> 3(> 145145 22(7 ' 1(> 22..4 4s 63 0~s4 123 11907 7222:29 o 3Th T2en5 9a1m EtGLn0a /*J~aG_8_m.~ 1178811s Z2a1is1m TsSceeann 120 2.55 8.81 5.06 6.82 7.57 8.88 Time Figure? Arepressive chiomtogramofa est simple (54.1032) Figure 7. A representative chromatograrn of a test sample (454-103-2). 22880000..00004422 M3MAA000022333998977 `WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDD -643.DIETAAPPPPPREEENPNDAIDRXIATITXVVION DIET PREPARATION PPRROOJJEECCTT NNOO..:" 445544--110033 WWeiegighhttaannddvvoolluummeeooffccoonsntsitittuueennttssuusseedd to0 pprreeppaarree teessttddiieests:: ConNNcoeonrmtmriaantalilons | Concentrations pom0ai (ppm a.i.) 1803 335616826..63 279113443.662 258963 1151787161 SubTTseetssattnce g Substance 0.18-18 00003..713628638138288 0211..9.744126628553299 58230 2.9123. 5.g239 1111.66448833 RBBaaatssiaaoll n 5R0a(0gtgi)0o0n 9888990999099909...6808 8888999999999998..3536 8888999099907784...1152 888999898848...244 DiDeiettsswweerreepprrepearepdaaassfrofollleloowwdss:: 5000.0gofbasal ration wasweighedinto taedHobartmixingbow. 5000.0 g of basal ration was weighed into a tared Hobart mixing bowl. TThhee tteessttssuubbssttaanncceewwaasswweieigghheedd iinnaattaarreeddwweieigghhbbooaatt.. Approximately 100gofbasal raion was ake romthemixingbowelandplacedin aWaring. blender. Approximately blender. 100 g of basal ration was taken from the mixing bowl and placed in a Waring TThheevtiteetshstttshsueubbrssittaannneccaeelwswaobasseaiadnddgdepelddattcooettdhhieebntblhleenendbdeleerrnaadnneddrt.thheewweeiigghhbbooaattwwaassrriinnsseeddwwiitthhaaddddiittioonnaall rraattiioonn,, with the rinse also being placed in the blender. <TThhee`bbblowlelen.nddTeerhrcecoobnlnteteennndttessrwwweearrseerbbillneesnnedddevddifftoohrrasadppdppirrotoxoxinmiamaltaeatletyliyo6n6,00wsiseetcchoonnhddesrsiaannnsddeattrrsaannossbffeeerrirrneegddttpooltathhceeemdmiinxixtiihnnegg mmiixxiibbnnooggwwl.l. The blender was rinsed with additional ration, with the rinse also being placed in the bowl. Thebowlwasplacedon aHobartmixerandthecontentsweremixedforapproximatelysixminutes "Theremainingration 2addedtothebowl andthe contentsweremixedfor six more minutes The bowl was placed on a Hobart mixer and the contents were mixed for approximately six minutes. The remaining ration as added to the bowl and the contents were mixed for six more minutes. Thedietwastransferredtoa labelledpaperfeed bag. The diet was transferred to a labelled paper feed bag. 22880000..00004433 33MMAA0000223393999988. : Eif 555885 $538582895AREY g EERE sssses sssssssssessss g 4 Ee 2 B53 ss5s%s sssssssessegsy| 2 : dE sss233 s3s3ssssessngy 3 Fs 5 8 |E ssssss sssssssegsssey i 3 , EL. i2 3 aul 88358 3558888%2882858 : g HgAl veer vegsrsgozzgses E[l0 Fite 58558 85855585855883 2 f HH d 5 4 [5 ssssss sssssssssssess Ef i S52 J& 2 of 58855 58825558855538 iF 25%) 2 5 3 Eg | oreo memamcmnanann [i3 : Z g 4 2 w Pooooood=sSE dRRERnEEnS E] IJJ 3 2 z 22880000..00004444 aMooz0asss 3MA00233999 WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLToD PROJECTNO. 454-103 PROJECT NO.: 454-103 a-4s5e- TEExperimental Group. Group Goma) (ppm a.i.) 0 0 o 0 AAPPPPEENNDDIIXX VVII InIdnidviviidduu`aAaclluBBtoeoddDyiyeW WtearieyigghThtotsxsi(5cg)i)ftfyrrooSmmtudaayNNwooirrttthhheePrrFnnOBSBo.bowbhwihtiete Acute Dietary Page | Toxicity Study Page 1 with PFOS Bird Bird DDsayy00 211 119 3323 2222110 Me545an 22zTan32n sDMean 221 SD 2 211 1i9 3423 F22Y200 4 5 222 sM MeDe5aann 22Ta12in SD 1 21 1 I19 332 3 20 0220 Me545an 2T22T22 sDMean 121 SD 1 211 1119 3+2 3 19 2119 Me45san T2221H2 soMean SD 120 1 DyS _ CDChhwaa0nnggSee Day 5 Day 0-5 xz2un4 s75 27 232 n11710 333392 111606 631 410 6 4 2226 757 Fy2259 1559 333 331] 19319 329 3 28 2 222%3 446+ 5222096 936 9 333550 11H38 529 38 5 3 2n8 99 5 5232070 1n118 EY33532 11 TM111 330 3 !10 1 DDaayys8 2327 "234024 35453900095 9 333555 Fi33357 43 3%39 338 3 3230 29 2532 "33470 534 5 3337 3233656 544E32] 339 4 CDhCaahyannsggsee Day 5-8 73 3 113700 11185 38 3 1909 0108s 18580 19 1 77 5 3373 755525 2 859 16 58 i0 1510 29 2 CTThoaountalgl e Change 158 2211114s 221i78s 718 7 111665 211"157 21 n717 217 2 1115 11292s 111358s 413 4 111368 16 2"17 21 2W21 219 2 22880000..00004455 33MMAA0000223344000000 WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDD PPRROOJJEECCTT NNOO.:.: 445544--110033 a6. - 46 - Teena Experimental Group. Group Gpmal) (ppm a.i.) o o GTooaw! AAPPPPEENNDDIIXX VVII InIdnidviviiddu"uaAalcluBBtoeoddDyiyeWWteaeriiygghThtotssx(i5(cg)i)ftfyrrooSmmtudaayNNoworirttthhhePerFnnBOBoSbowbhwihtiete Acute Dietary Pags2 Toxicity Study Page 2 with PFOS BBiirrd Dayo Day 0 211 21199 2 3 s3 20 2221 Me455an 2222200 soMean SD 120 1 21 1 2118 4235 21 2119 Me455an 2222120 sMMSeDeDarn 220022 soMean SD 120 1 Days ClDCuhnna0nggsee Day 5 Day 0-5 P 5 28 9 7 7 27 7 2 i 32 11 0 1 30 10 a n 34 12 0 0 30 10 3 2 3 2 0 n 30 12 3s 1 35 14 10 29 10 n 32 11 2 0 32 10 2 i 32 11 2 2 2 2 0 10 30 10 4 5 4 3 DDauys8 333s5 36 "-44 4u4 s40 5 339 %46 337 a41 242 a41 33 3%38 55 DCCruhaasnnggsee Day 5-8 7 79 122 9 11010 210 2 n99 15918 115900 5911 28 2 CTThoaountalgl e Change 111666 16 3-23 2ni2o 419 4 2221 22i158s Fa2200y 2T133S 418 4 22880000..00004466 33MMAA0000223344000011 WILDLIFE INTERNATIONAL, tid WILDLIFE INTERNATIONAL, LTD PPRROOJJEECCTT NNOO..: 445544--110033 a1 - 47 - AAPPPPEENNDDIIXX VVIl IInnddiivviiddu`uAaaclluBBteoodDdiyyeWWtaereiyigghThtotsssi((g2c))iyffrroSomtmudaayNNwooirrttthhheePrrRnnOBBSoobbwwhhiittee Acute Dietary Page Toxicity Study with PFOS Page 3 eee------------------------------------------------------------ ExGppGGerrmrioomauuieppn)tal Bid Dwy0 Days CChDhayann0ggSee Days CDhCahaanynggsee ChTTaoontalgle (ppm a.i.) Bird Day 0 Day 5 Day 0-5 Day 8 Day 5-8 Change 183 1 1 % 7 2 5 15 18,3 1 19 26 7 34 8 15 2 1 5 " 6 2 19 33 14 39 6 20 3 2 n 6 3s 5 u 3 21 27 6 35 8 14 soz 4 2 " n 4 22 34 12 46 12 24 son 56 " " 1 5 22 36 14 46 10 24 Man TTT 90 Mean 21 31 11 40 9 19 ) 2 4 4 2 5 SD 2 4 4 6 2 5 183 18.3 Gop Group Toul Total 1 1 5 38 0 1 1 19 28 9 38 10 !9 2 1 s 5 5 2 19 28 9 37 9 118 32 3 9 1 3 20 29 9 38 9 18 soz 5 n " 0 2 4 22 34 12 44 !0 22 so 3s B " 5 zn 5 22 35 13 44 9 22 M MsaeDann TW H202 3313 120 R 430 91 2B02 EM MSoeeDa)nn 23112i 3 3431 2 i311 3 540 529 1 2 7320 SD 1 4 3 5 2 3 228800001.00004477 33MMAA0000223344000022 WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LL-1o-D a. - 48 - PPRROOJJEECCTTNNOO:.: 445544--110033 Individual Body WeiAAghPPtPPsEE(NNg)DDfIIrXXomVVIaINorthem Bobwhite IndividAAucaculutBteeoDdDiyieeWttaaerryiygTThotoxsxi(icgci)ittyfyroSSmttuuddayyNwwoiirttthhhePPrFFnOOBSSobwhite PPaaggee 4 --_---- eee -- ExGGpprGemrorioamwuipe)ntal Bid Do (ppm a.i.) Dus ChDCawhna0ngsge_e Dws _ DCChhuaasnnggsee CTThoaountaglle Bird Day 0 Day 5 Day 0-5 Day 8 Day 5-8 Change 366 36.6 --_-- 11 1 18 21 2 19 3 3 119 4 + 221 s 5 221 sMMeDeamn TH201 SD 1 2 33 9 26 8 35 9 17 30 un Ii 339 9 9 2 229 3 8 20 1s 10 37 53 8 5 n18 30 9 38 8 3 5 ws 9 217 34 13 43 9 22 3 W3w05 W 2 120 m 3 385 5 0 91 m a 192 3 2 3 1 2 368 36.6 1 1 1 2 10 Ed 8 1 2 19 29 10 2 10 37 Ed 2 8 5 19 518 32 =29 n10 37 8 8 3 18 21 +2 32 3s 111 "40 8 5 219 22 35 13 44 9 5 36 n" s 222 23 36 13 44 8 21 _-- sMeDm Tm A212 325 112 0 4W+w0 08 w2m02 L TTGGoorouotauplp MMaSsSeDDDann 2 220222e 5 3311333 21W 212 3 m3b 349 a 08s 1 2 h1922 2 3 1 2 22880000..00004488 33MMAA0000223344000033 W WIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDD - 949.- PPRROOJJEECCTT NNOO.:.: 445544--110033 =---- Experimemal "Group, Group @pmai) (ppm a.i.) 273.2 AAPPPPEENNDDIIXX VVII IInnddiivviidduu`aAalcluBBteooddDyyiaW WereiyigghThtotsxsi((cggi))tfyfrroSomtmudaayNNoworirttthhheePrrFnnOBBoSobbwwhihittee Acute Dietary Pages Toxicity Study Page 5 with PFOS Bid Bird Dyo Day 0 1 1s 1 18 2 1 2 19 3 3 20 4 2 4 21 s 2 5 21 Mew Mean 20 sD 1 SD 1 Days CChDhaawnn0ggSee Dns CDChuhaaynn3gg8ee Day 5 Day 0-5 Day 8 Day 5-8 CTThooataaml e Change 226 3g 232 56 114 226 73 7 33 7 7 114 229 9 5 336 7 7 1166 232 1uw1 339 7 7 is 18 30 9 ' 3% 36 5 6 115s T2B93H s 91 m 353 7 71 6125 3 1 3 1 2 752 1 1 1185 z27 59 334 77 1166 .73.2 2 2 3 u 5s 7 1s 2 20 31 11 38 7 18 3 2 20 99 337 7 7 16 16 3 21 30 4 221 232 n11 339 77 1185 4 s2 2 0 a 5 22 32 10 41 9 MMseeDann 22200 2320 1101 33838 771 19 n117 TGoGruroolwup MMsSeeDDaan 220201 33022 59 1 1 337373 7711 T162o1 Total SD 1 2 1 3 1 2 22880000..00004499 33MMAA0000223344000044. W WIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDO --5s00-- PROJECTNO: 454-103 PROJECT NO.: 454-103 AAPPPPEENNDDIIXX VVII IInnddiivviiddu"uAaaclluBBteoodDdiyycWWteaieriygghThtotssri((g2c))iyffrroSomtmudaayNNwooirrttthhhePerFnrOBBSoobbwwhhiittee Acute Dietary Pago Toxicity Study with PFOS Pago 6 rs Experimental GGrpomuapi.) Group Did Dny0 Dns ChDCahnnangsgee Dys CChDhauanyngsge8e CTThooautnalle (ppm a.i.) Bird Day 0 Day 5 Day 0-5 Day 8 Day 5-8 Change 16 11 119 28 9 335 17 116 146 21 2 4 28 9 32 4 B13 2 -19 sow u 4 . - - 3 + 20 24 oz El 4 n 3 s 3 s 5 4 22 33 511 238 5 ` 16 5 MMe)eann 23 28 5 32 4 9 E 2 5 3 m 3 3 1 W3 L 21 28 8 34 14 SD 2 3 3 3 3 us 1 1 u 6 a 7 5 146 1 18 24 6 31 7 2 1 7 3 3s 5 16 2 19 27 8 35 8 3 1 7 5 El " 3 19 27 8 33 6 a - - - - - 4 21 5s 0 u 1 EY ` s 5 23 24 1 28 4 Gow EMMoee)ann Tm202 E2d 6T 3 w=3 I2 T5e Group Toa! 5) 2 3 5 3 2 : Total Mean 20 26 6 32 6 SD 2 2 3 3 2 Mean 20 27* 7* 33* 6** SD 2 3 3 3 2 Emme (-) = No data available due to mortality. Sisal differfom he contl groupsp< 0.05(Dimmers est). *Statistically different from the control group at p < 0.05 (Durmett's t-test). Stuiscaly ifn Gotheeatsc ros L001(Don's sh) **Statistically different from the control group at p < 0.01 (Dmmett's t-test). 13 16 14 5 12 5 13"* 4 22880000..00005500 33MMAA0000223344000055 WWIILLDDLLIIFFEE II NNTTEERRNNAATTII OONNAALL,, LLTTDD -s5i1 e- PPRROOJJEECCTT NNOO.:.: 445544--110033 AAPPPPEENNDDIIXX VVII IInnddiivviidduu`AaaclluBBteooDddiyyetWWaereiyigghThtotxssi((cggi))tyffrroSomtmuadaNyoNwroitrtthhheePrrFnOBBSoobbwwhhiittee Acute Dietary Page Toxicity Study Page 7 with PFOS ------------------------------------------ Experimental GGrmoauipl) Group Bid Dyo Dys ChCaDhnwangOgSse Duns ChCDahuanyng3gese CTThaoataalle (ppm a.i.) Bird Day 0 Day 5 Day 0-5 Day 8 Day 5-8 Change 2293 Tom 2 1 18 20 2 2 221 ' 5 21 n 2 - - . 2 19 FI) 117 2-2 - . . 3 20 + on -18 --2 - - . 4 22 som 0 2 - . . 5 22 20 -2 Maw Mean w 202 TH2T T 21- : : SD 2 Ee 1 1 1 o 293 1 19 19 0 rw . - 2 19 sow 2 o 3 20 20 0 sow w 3 4 20 17 -3 som 1 5 5 21 16 -5 Gop MMeaawn TH201 DW 2E Z 2 Group Toa 3 1 2 2 Total Mean 20 SD 1 Mean 20 SD 1 18 2 18"* 2 -2 2 -2** 2 ER(-) = No data available due to mortality. Sinica ile oe he contdgo tp < 005 Due' 0) *Statistically different fi-om the control group at p < 0.05 (Dunnett's t-test). "Suisealydlls rn he cont ro tp 001 (Desist). **Statistically different front the control group at p < 0.01 (Durmett's t-test). 1s + " -15 .-4 e - - - - - - - -- - - B 15: F-42 A: - 418"* 4-2** -E1s"e* 4 4 -- 5 22880000..00005511 33MMAA0000223344000066 WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDD 52- -52- PRPROOJJEECCTTNNOO.:.: 445544--110033 AAPPPPEENNDDIXIVXI InIdnidviviidd"uuAaaclluBBtoeoddDyyiWtWeairegyighThtotssx(i5g)i) yffSrrooimmndaayNNwoiorrttthhheePrrnFBOBoSoobwwihiitte Acute Dietary Pag8e Toxicity Study Page 8 with PFOS Tepermenael e Experimental GGortou)p Group (ppm a.i.) st6 586 BiBird DDawyo0 1s 21 m18w 2som20 s3 om21 sm 4 21 MMeD e5aann 22T220H SD 2 Dus CrCahDnawnggsee Day 5 "Day 0-5 5. a- 13 -7 3 18 -3 . - - " un 5 17 -5 i 5 16 -5 3 2 3 2 bus ClCaDhnaynsggtee Day 8 Day 5-8 CTTohoutamll Change -. . -. . -. . -- - 3 S-- M . > : : 555 rw 3 a - ' . 586 19 16 -3 2 0 - - - - - 2 20 " sw 3 4 - - - 3 20 16 -4 sm - - - - . 4 19 som . - - ' . 21 GTooupl M[ S eno-- ir 30 ie 2 a=2 =a "g: 7&: Group Total Mean 20 SD 1 Mean 20 SD 1 16 0 16"* 2 -4 1 -4"* 2 C25 EiE tera bR o col ropa <001 Dost es) (-) = No data available due to mortality. **St_~ti~tically different from the control group at p < 0.01 (I)l~nett's t-test). 22880000..00005522 3MMA0A000223344000077 WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTDD se -53 - AAPPPPEENNDDIIXX VVII InIdnidviivdidu"uaAalcluBBtoeoDddiyyeWtWeairegyihgTthostvs(i(2cgi))tfyfrroSomtmuadayNNwoiortrththheePrrFnBOBoSbowbhwihtitee Acute Dietary Pages Toxicity Study Page 9 with PFOS PPRROOJJEECCTTNNOO..;: 445544--110033 Experimental pcGroup Gpmai) (ppm a.i.) n11m71 Bid Dayo Bird Day 0 cage Diys Day0s Day Day 5 Chazzge Day 0-5 Day 8 Dayss Chm~ge Day 5-8 ----Change Total Change ! 1s - . - . . 1 18 ER) - - - , . 2 20 sow - - - , 3 20 om - -- - . 4 21 soa - -- . Man5 Mean T 22110 : H : : T : : SD 1 wm 1 1s - - - 1171 1 18 - - 2 ow - = - 2 20 - - 3 ow - - - 3 20 - - + =n - - - 4 22 - som . - - 5 21 - BEMean 20 1 : - - SD 1 Gow Mean 20 B : 5 Group Mean 20 Total 1 - : Total SD 1 RETR (-) = No data available due to mortality. . - . o - . . . 3 ---::fen :> - 22880000..00005533 33MMAA0000223344000088 W WIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LL-TrDO san - 54 - PPRROOJJEECCTT NNOO:.: 445544--110033 EExxGppereroriiummepen.nttaall pmGroup (ppm a.i.) 0 0 0 0 GTGoorouwulp Total AAPPPPEENNDDIIXX VVII IInnddiivviidduu"AaaclluBBteooDddiyyetW WaereiyigghThottvssi((cggi))tyffrroSomtmudaayNNwooirrttthhheePrrFnnOSBB,oobbwwhhiittee Acute Dietary Page 10 Toxicity Study Page 10 with PFOS BBiirdd DDauys8 21 1 3z2n7 +253 "243024 Me455an 40 9050 sDMean 539 SD 9 211 333ss5 .233 35 5337 4 5 343 sM MeDe5ann 33T983E SD 3 211 3.3200 hl323 29 3=32 Me545an "433078 MMseSeDaDann 33475s soMean 537 SD 6 DDauyyi1s5 242 s656 &69 a63 778 a62 114 s55s5 k&65A34 974 59 a361 8 si5ts4 5545554 5a56558 E5]585 I5o9 10 ChCDahynasngglees Day 8-15 2115s 22 3227 222223385 5 220 i]18 227 331 220 223 66 22146 16 2i22 18 2228 B2r525 523 5 Dy Day 22 a671 77 3292 11a080444 1684 16 77744 317085345 2103 80 wn83 12 779 63 n77 275 995 378 I11 5)82 113 CDlCaaytmninsggzee: Day 15-22 1919 221 223 221 226 222 33 119 221 221 229 221 222 44 225 1s18 523 220 727 723 34 722 33 CTThoaonutaglle Change p33H4 4u55300 as45st44 545 8 33399 39 &848 a60 i449s15 9 3a49 3r345l4 38 "555 w44s45538 8 22880000..00005544 33MMAA0000223344000099 W WIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LTD LTD Lss- -55 - APPENDIX VI IInnddiivviiddu`uaAallctBBsooddDyyieWWteaeiriygghThtotssxi((cg8i))tffyrooSmtmudaayNNwooirrttthhhePerFnnOBBSoobbwwhhiittee Acute Dietary Page 11 Toxicity Study Page 11 with PFOS PPRROOJJEECCTTNNOO:.: 454-103 454-103 TEexGpreoprwimn,eental @pnal) Group (ppm a.i.) BdBird Dus Day 8 183 11 u34 18.3 23 2 39 5 3s 3 35 + a6 4 46 55 %46 MMseeoawn 4W05m SD 6 Dyls CDhCuahyaninglgees Day 15 Day 8-15 s 1 53 19 6 2% 65 26 2s 60 25 3 z 73 27 n 2 71 25 &@ 64 24 5 5 8 3 own Day 22 Ed 78 8s 85 58%6 9 95 292 8w77m 7 Change Change yim Day 15-22 225 20 226 222 221 2B33m 3 cTounl Total Change " [346 551 @49 a46 4a7 3 3 _--Experimental Ex(pGGperrmroiomauupiepntal Bid Day8 Dayls CDhCaahynagnglees. Day22 DCaChyhaalnnsgge2e2 (ppm a.i.) Bird Day 8 Day 15 Day 8-15 Day 22 Day 15-22 TCToootatamalgl e Change 66 11 33s5 060 225 8811 221 4%6 36.6 2 6 2% 9 22%6 5522 2 39 65 26 91 3 Ed 6 2% 9 2% 5s4t 3 37 65 28 91 26 4 3 6 = 5 2 a45 4 5 38 a 61 7 23 83 22 o7 2 5s4t 5 43 72 29 97 25 MMSeeDan.n 33 83 6& 55 262288%97 # 224 0503 SD 3 5 2 7 2 4 22880000..00005555 33MMAA0000223344001100 WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LtZTTOD -56- - 56 - PPRROOJIEECCTTNNOO.:.: 445544--110033 --_E--xGpr--eori--umpental Gpmai) Group (ppm a.i.) 7273.2 AAPPPPEENNDDIIXX VVII IInndciivviidd"uuAaaclluBBteoodDdiyyeW WtaereiyigghThottsssic((iggt))yffrroSomtmuadayNNwooirrttthhheePrrFnnOBBSoobbwwhhiittee Acute Dietary Page 12 Toxicity Study Page 12 with PFOS Bid Days Dls CDhCaahyannglgees Dy CDhCaahyanlnggseez Bird Day 8 Day 15 Day 8-15 Day 22 Day 15-22 _TTCahoataalml e Change 1 1 232 s5t4 222 %76 222 "44 2 5 EY 2 nom 77 19 "44 2 33 5 3% @58 %25 on a 3 ` 336 662 326 7w9 oz17 43 E) 4 s 3369 565 26 zn 7 1 87 22 @48 5 M M(a5en)nu 36 59 23 78 19 42 Tm H5 4 @2 w 4 2 #@2 35 60 24 79 20 44 SD 3 4 2 4 2 2 EpxGpererorimumpenetal GpGuraolup) Bid Duy8 Duis CDhCaahyansngglees Dwy2 CDhCaahynaigngsse.z (ppm a.i.) Bird Day 8 Day 15 Day 8-15 Day 22 Day 15-22 CTThoaountalgl e Change 146 ! 5s EY 2 a 5 81 23 " 46 146 1 35 58 23 2 2 El 2 nz as 2 32 55 23 77 22 45 3 - ` 3 @ 2 5 1 " 4 38 63 25 81 18 43 s ES 5 2 wa a 5 32 57 25 78 21 46 Mw m WW ma BL Mean 34* 58 24 79 21 45 sD 3 3 1 2 2 1 SD 3 3 1 2 2 1 Nod el Bom (-) = No data available due to mortality. Smita ie rom hclgop tp < 005 (Duetsst) *Statistically different from the control group at p < 0.05 (Durmett's t-test). 22880000..0000,5566 3MMAA00Z032434001111 W WIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDO 57. -57 - PPRROOJJEECCTTNNOO..:: 445544--110033 AAPPPPEENNDDIIXX VVII IInnddiivviidduu`AaaclluBBteooddDyiyeW WteaeiriygghThtotssxi((cggi))tffyrroSomtmudaayNNwooirrttthhhePerFrnOBBSoobbwwhhiittee Acute Dietary Page 13 Toxicity Study Page 13 with PFOS = Group, Experimental Change (ppmGroup (ppm a.i.) Bid Bird DDaayyo0 DDaayys5 Dayos Ctmnge Day 0=5 DDawy g8 DCChhaaaynnggsee Day 5-8 ChTTaoonutgalel . Change 22993 1 1 221 335 114 5555 020 u34 2 - - - - e . 2 5 -- . . - e . 3 4 - - . - - . . 5 - . - . . . 5 . Maw E Wm B mW Mean 21 35 14 55 20 34 sD . : i . : SD - - Team (-) = No data available due to mortality. 22880000..00005577 33MMAA0000223344001122 W WIlLLDDLLII FFEE IINNTTEERRNNAATTIIOONNAALL,, LLTfDD PPPR.OOJJEECCTT NNOO:.: 445544--110033 -ss- -58 - AAPPPPEENNDDIIXX VVIII FFeecedd CCoonnssuummptpitioonDnig(cgbt!iabrirrdydd/Tadoyaxy)i)cbbiyytPyPeSentnuffdrryoommwitaahNNPoorFrttOhhSeerrnnBBoobwbhiite AAccuuttee Dietary Page 1 Toxicity Study Page 1 with PFOS oem Go Experimental Group Opmai) Pa (ppm a.i.) Pen EExwpoosuwrenPfeeriodd | Day D0w-O5s FoBpousFed Pest-Exposure Period DDwaey 6s-8 Conral Control 1 1 9 s 212 2 2 7 7 212 5 3 8 s ?7 4 n 4 12 212 5 5 5 9 9 ' 66 010 9 ' MM)eman T 29 y w12m0 SD 2 2 22880000..00005588 M3MA0A002023344001133 W WIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDD PPRROOJJEECCTTNNOO.:.: 445544--110033 -59- AAPPPPEENNDDIIXX VVIIII FFeeeedd CCoonnssuummppttiioonnD(i(bget/ibarirrddy/dTdaoayxyi))cbbityyyPPSeennudrfryoommwitaahNNPoorFrttOhhSeermn BBoobbwwhhiittee AAccuuttee Experimental Gop Dietary To:deity Study with PFOS EPxapogseu?rs Period Page 2 Post Exposure Fol E~erimental Group E~osure Period Post-Exposure Period G(pppmmaai.i).) rPean TDT mDayo 0-5 s Dm Day 6-8 183 18.3 21 1 599 1u10n0 Mea2 n 58 1i1T Mean 9 11 366 36.6 211 188 514 Mea2 n 7 5 79 Mean 8 12 na 1 10 73.2 1 10 12 2 10 1 2 10 14 Mean 0 Mean 10 13 16146 21 1 1077 158 Mea2 n Mean 510 9 To12 10 2293 21 1 4 4 Me2n 36 Mean 5 st586 21 1 `6 5 Mea2 n <5 Mean 6 mm 1 4 1171 1 4 2 4 Mean T Mean 4 Toma Ey - No data available due to adult mortality. 18 5 510 9 116 222 i19 . A-SA 22880000..00005599 M3MA0A002023344001144 WILDLIFE INTERNATIONAL, LTD WILDLIFE INTERNATIONAL, LTD PROJECT NO: 454-103 PROJECT NO.: 454-103 60- - 60 - AAPPPPEENDNIDXIVVIXII FeFeeeddCCoonnssuummptpitioonnD(i(gcgbt/ibarirrdyd/T/ddaayyo))bbxyyPPSeieunndffcyroowmmiitaahNNtoProFtryOthheSerrnnBBoobbwwhihtiteeAAccuuttee Dietary Pages Toxicity Study Page 3 with PFOS EemenatGon Experimental Group Gomai) (ppm a.i.) Beowsted | PowBpeuered Exposure Period mT wEs CT twBE Pen Day 8-15 Post-Exposure Period Day 15-22 Control Con~ol 1 1 2 2 5 EM M)ee3 awn SD 0 1 10 14 0 10 12 v 2 7 12 3B 9 13 2 i 2 1 me Gop Experimental Group Goma) (ppm a.i.) Pen 183 1 18.3 366 1 36.6 m2 1 73.2 6 1 146 253 1 293 Epeerad Exposure Period Day 8-15 10 10 u14 513 u11 5 Petite Post-Exposure Period 7 Day 15-22 212 15 15 1s 15 u14 5 22880000..00006600 M3MA0A002023344001155 W WIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDD PPRROOJJEECCTT NNOO..:: 445544--110033 --6611-- AAPPPPEENNDDIIXXVVIIIIII CCHAHNGAESTTNOOPPGRROOTETOOCCSOOLL 1w.1a.TsThhceleparrpiorfotioteocdtcoololwiwnadasiscaaamtmeetenhndedecedodtltlooeicintndidoiniccaaottfeetlhtihavaetrtbibflirleeowmwiblilillrbdbesectochloaltlledeccitteeedddfdfrurrooimmnaglatllhlsesttuucddoyyubrbisirerddossft.. ThTehhseetpuprdroyot.tooccooll was clarified to indicate the collection of liver from birds that died during the course of the study 2t2u..BbBelslo.ooToddhsseaapmmrpopltleeosscowwleerirenedcicocolalllteeeccdttteehddaotonnhDeDpaaayryi n88aiaznnedddvDDaauayyt2a2i2nieinnrnsnoownn-o-huhelepdpabareriinunisizzeeeddd.5$mmllbboororosisliliiccaattee lasstest glass test tubes. The protocol indicated that heparinized vacutainers would be used. P33.p. TThaheep,prroo0tto0occ,ooll1w8wa.a3s,saa3m6me.e6n,nd7de3ed.dt2t,ooc1ch4h6aa,nn2gg9oe3tt,hheet58tee6tsatcncodoncn1ec1net7nr1tpraatptiimoonnsas.fifr.roommTe0s0,t,c2200o,,44n00,c,880e0,,n1166t00w,,re66r4a4e00ctahanaindndog11e2n2d88ts00o rpefplmecat.ti.h,etotes0t,su1b8s.3t,a3n6c.e6p,ur7i3t.2y,g1i4v6e,ni29t3h,e5n8e6wacnedti1f1i7c1a ppomfaan.ai.lysTiess.t concentrations were changed to reflect the test substance purity given in the new certificate of analysis. n44o,.tTThrheeectoteermmdpeepdrearattuurreessffrroommsesveevrearlalbbrrooooddeerruunniitssoonnDDaayy44,, 77,, 88,, aannddaalll bbrrooooddeerruunniittssoonnDDaayy2222wwereree. not recorded 5t5..TThrheeaafefttgeerrrnnoaoouoopnntoanobbsAmesperrrvivaelat2tii6oo,nnnss19wot9er9ro.e inadvertently inadvertently notrecordedfor2birdsinthe640ppm not recorded for 2 birds in the 640 ppm 2.1 a.i. treatment group on April 26, 1999. 22880000..00006611 33MMAA0000223344001166. Lo ' WWIILLDDLLIIFFEE IINNTTEERRNNAATTIIOONNAALL,, LLTTDO -"6622APPENDIX IX APPENDIX IX PERSONNEL INVOLVED IN THE STUDY PERSONNEL INVOLVED IN THE STUDY PPRROOJJEECCTT NNOO.:.: 445544--110033 "TThhee ffoolllloowwiinnggkkeeyyWWiildliife IInntteernrantiaotnailoLLndtad..ppeerrssoonnnneellwwerereeininvvoollvveeddiinntthheeccoonndduucctt oorr mmaannaaggeemmeennttoofftthiiss ssttuuddyy:: 1)(1) MMaarrkk JTaabbeerr,, Wii Wildlie Toxicaogis Toxicologist 0) JJooaannnn'BB.. BBeeaavveerrss,, DDiirreeccttoorr,, AAvviiaann TTooxxiicccollooggyy ((23)) Sean P. Gallagher, Senior Diclgist ((3@)) CSocuarntPo.eyGCaallsaegyh,erM,.SS,eniSoraBiisoloBigaisotgist: ((45)) WCiolulratrndeyB.CNa-iqxeoyn,,MP.hS..,DS,eMnaionraBgieo,loAgnisatlytical Chemiscy ((56)) W TiilmlaortdhBZy..NKiexnodna,lP,hS.Dup.e,rMviasnoarg,eArn,aAytniaclyatlicCalhCmihsemeyistry ((67)) RaymonLd. Van Hoven, PhD, Timothy Z. Kendall, Supervisor, Raymond L. Van Hoven, Ph.D., Sietist Analytical Scientist Chemistry )(7) Ellen Mak, Chemist (8) Ellen Mank, Chemist 22880000..00006622 mA00234017 3MA00234017