Document Z4y8kYQL8Y14QRD0mZ3vyNMVV

-1 Perfluorooctanesulfonate: Complete Cross-Fostering Study in Rats + Serum Analytical FINAL REPORT PROTOCOL 418-014 ORAL (GAVAGE) CROSS-FOSTERING IN RATS STUDY OF PFOS SPONSOR'S STUDY NUMBER: T-6295.13 FINAL REPORT DATE-:-23 JULY 1999 PROTOCOL 418-014 ORAL (GAVAGE) CROSS-FOSTERING IN RATS STUDY OF PFOS SPONSOR'S STUDY NUMBER: T-6295.13 TABLE OF CONTENTS SUBJECT 1. SUMMARY AND CONCLUSION A. Methods B. Results C. Conclusion ii. DESCRIPTION OF TEST PROCEDURES A. Conduct ofStudy A.1. Sponsor A.2. TestingFacility A.3. Study Number A.4. Sponsoes Study Number A.5. Purpose ofthe Study A.6. Study Design A.7. RegulatoryCompliance PAGE 1-1 1-1 1-4 1-6 11-1 11-1 11-1 11-1 11-1 11-1 11-1 11-1 11-1 SUBJECT A.8. Ownership ofthe Study A.9. Study Monitor A.10. AlternateStudy Monitor A.11. Study Director A.12. TechnicalPerformance A.13. Report Preparation A.14. Report Review A.15. Date ProtocolSigned A. 16. Dates ofTechnicalPerformance A.17. Records Maintained B. Test ArticleInformation B. 1. Description B.2. Lot/BatchNumber B.3. Date Received and Storage Conditions B.4. SpecialHandling Instructions B.5. AnalysisofActivity C. VehicleInformation C. 1. Description C.2. Lot Numbers C-3. Dates Received and Storage Conditions C.4. SpecialHandling Instructions C.5. Analysisof Purity ii PAGE 11-2 11-2 11-2 11-2 11-2 11-2 11-2 11-2 11-3 11-3 11-3 11-3 11-3 11-3 11-4 11-4 11-4 11-4 11-4 11-4 11-4 11-4 SUBJECT D. Test ArticlePreparationand Storage Conditions D. 1. Sample Information D.2. AnalyticaRlesults E. Test System E.1. Species E.2. Strain E.3. Supplier(Source) E.4. Sex E.5. RationaleforTest System E.6. Test System Data E.7. Breeder Male Rat Data E.8. Method of Randomization E.9. System of Identification F. Husbandry F.1. Research FacilitRyegistration F.2. Study Rooms F.3. Housing F.4. Lighting F.5. Sanitization F.6. Feed F.7. Feed Analysis F.8. Water PAGE 11-5 11-5 11-5 11-5 11-5 11-5 11-6 11-6 11-6 11-6 11-6 11-6 11-7 11-8 11-8 11-8 11-8 11-8 11-8 11-8 11-9 11-9 I SUBJECT F.9. Water Analysis F.10. Bedding F.11. Bedding Analysis G. Methods G.l. Dosage Administration G.2. RationaleforDosage Selection G.3. Route ofAdministration G.4. RationaleforRoute ofAdministration G.5. Frequency ofAdministration G.6. Length ofStudy G.7. Method ofStudy Performance H. Gross Necropsy H. 1. Fo Generation Rats H.2. Fl GenerationPups 1. StatisticAanlalyses Ill. RESULTS A. MortalityC,linicaalnd Necropsy Observations B. Body Weights C. Absolute(g/day)and Relative(g/kg/dayF)eed Consumption Values D. NaturalDeliveryand LifteOrbservations D-1. NaturalDeliveryObservations D.2. Cross-FosteringLifteOrbservations iv PAGE 11-9 11-9 11-9 11-10 11-10 11-10 11-11 11-11 11-11 11-11 11-11 11-15 11-15 11-16 11-17 111-1 111-1 111-1 111-2 111-2 111-2 111-2 SUBJECT E. ClinicaOlbservationsfrom Birthto Day 21 Postpartum and Necropsy Observations F. ElectronMicroscopicEvaluationof the Liverand Lung from Day I Postpartum Pups F.1. Liver F.2. Lung REFERENCES APPENDIX A - REPORT FIGURES PAGE 111-3 111-4 111-4 111-4 111-5 Figure1. Matemal Body Weights A-1 Figure2. Matemal Body Weights - Rats Assigned toCross Fostering A-2 APPENDIX B - REPORT TABLES Table 1. Table 2. Table 3. Table 4. Table 5. Table 6. Table 7. Table 8. ClinicaOlbservations- Summary - Fo Generation Female Rats B-1 Necropsy Observations- Summary - Fo Generation Female Rats B-4 Body Weights - Precohabitation-Summary - Fo Generation Female Rats B-5 Body Weight Changes - Precohabitatio-nSummary - Fo GenerationFemale Rats B-6 Matemal Body Weights - Gestation- Summary - Fo Generation Female Rats B-7 Matemal Body Weight Changes - Gestation-Summary - Fo Generation Female Rats B-8 Matemal Body Weights - Lactation-Summary - Fo Generation Female Rats B-9 Matemal Body Weight Changes - Lactation-Summary - Fo Generation Female Rats B-10 v SUBJECT Table 9. AbsoluteFeed Consumption Values (g/day)Precohabitation-Summary -Fo Generation Female Rats PAGE B-1 1 Table 10. RelativeFeed Consumption Values (g/kg/day)Precohabitation- Summary -Fo Generation Female Rats B-12 Table 11. Matemal Absolute Feed Consumption Values (g/day)- Gestation-Summary - Fo Generation Female Rats B-13 Table 12. Matemal RelativeFeed Consumption Values (g/kg/day-) Gestation- Summary - Fo GenerationFemale Rats B-14 Table 13. Matemal AbsoluteFeed Consumption Values (g/day)- Lactation- Summary - Fo GenerationFemale Rats B-1 5 Table 14. Matemal RelativeFeed Consumption Values (g/kg/day)Lactation- Summary - Fo GenerationFemale Rats B-1 6 Table 15. NaturalDeliveryObservations- Summary - Fo Generation Female Rats B-1 7 Table 16. LifteOrbservations(NaturallDyeliveredPups) - Summary - Fl GenerationLifters B-20 Table 17. ClinicaOlbservationsfrom Birthto Day 21 Postpartum - Summary - Fl GenerationPups B-24 Table 18. Necropsy Observations- Summary -Fl GenerationPups B-25 Table 19. ClinicaOlbservations- IndividuaDlata - Fo Generation Female Rats B-26 Table 20. Necropsy Observations- IndividuaDlata - Fo Generation Female Rats B-30 Table 21. Body Weights - Precohabitation- IndividuaDlata - Fo GenerationFemale Rats B-33 Table 22. Matemal Body Weights - Presumed Gestation- Individual Data - Fo Generation Female Rats B-45 Table 23. Matemal Body Weights - Lactation-IndividuaDlata - Fo Generation Female Rats B-53 vi SUBJECT Table 24. Feed Consumption Values -Precohabitation- Individual Data - Fo GenerationFemale Rats PAGE B-59 Table 25. Matemal Feed Consumption Values - Presumed Gestation- IndividuaDlata - Fo GenerationFemale Rats B-63 Table 26. Matemal Feed Consumption Values - Lactation- Individual Data - Fo GenerationFemale Rats B-71 Table 27. NaturalDelivery,ImplantationSites,and Pup Viabilitaynd Sex - IndividuaDlata -Fo Generation Female Rats/ Fl GenerationLifters B-74 Table 28. DurationofGestationand Average Durationof Delivery (Minutes)Per Pup Per Lifte-rIndividuaDlata - Fo GenerationFemale Rats B-77 Table 29. Pup Body Weight LifteArverages from Birthto Day 21 Postpartum - IndividuaDlata -Fl Generation Lifters B-80 Table 30. Pup Body Weights from Birthto Day 21 Postpartum - IndividuaDlata - Fl GenerationPups B-83 Table 31. Pup VitalStatusand Sex from Birthto Day 21 Postpartum - IndividuaDlata - Fl GenerationPups B-99 Table 32. ClinicaOlbservationsfrom Birthto Day 21 Postpartum IndividuaDlata - Fl GenerationPups B-1 02 Table 33. Necropsy Observations- IndividuaDlata - Fl Generation Pups B-1 04 APPENDIX C - PROTOCOL AND AMENDMENT C-1 to C-41 APPENDIX D - DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY D-1 APPENDIX E - TEMPERATURE REPORTS AND RELATIVE HUMIDITY E-1 to E-7 APPENDIX F - PATHOLOGY REPORT F-1 to F-46 APPENDIX G - STATEMENT OF THE STUDY DIRECTOR G-1 vii SUBJECT APPENDIX H - QUALITY ASSURANCE STATEMENT UNIT FINAL REPORT PAGE H-1 to H-5 viii 418-014:PAGE I- TITLE: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS ARGUS RESEARCH LABORATORIES, INC. PROTOCOL NUMBER: 418-014 ';PONSOR'S STUDY NUMBER: T-6295.13 1. SUMMARY A. MethodSa AND CONCLUSION Seventy-eight virgin female CH:CD@BR VAF/Plus@ (Sprague-Dawley) rats were assigned totwo dosage groups (Groups Iand 11)4,2 ratsinthe control group and 36 ratsinthe treatedgroup. The ratswere administeredthe test articleP,FOS (FC-95),orthe vehicle0,.5% Tween@ 80, orally(viagavage),once dailybeginning42 days beforecohabitatiotnhroughday 21 postpartum(DL 21). The dosage volume was 5 mL/kg. Dosages were adjusteddailyforbody weight changes and given atapproximatelythe same time each day. Rats were assignedto cross-fosterinogr pharmacokineticsample collection depending on the orderof deliveryand the availabiliotfyratsforcross-fostering. Twenty-fivelifterpser dosage group were assignedto cross-fostedngon DL 1. The lifterwsere assigned such thatfoursubsetswere obtainedamong the two dosage groups as describedinthe tablebelow. Dosage Group 1 1 11 L 11 Number of Mated Female Rats 13 12 12 13 Dosage (mg/kg/day) 0 (Vehicle) 0 (Vehicle) 1.6 1.6 Number of Litters Reassigned 13 12 12 13 Reassigned Litter Type Subset (Dam Treatment) Number Treated A Untreated B Treated c Untreated D Eightfemale ratsfrom the controlgroup and two female ratsfrom the treated group were assigned to pharmacokineticsample collectiofnollowingdelivery. These ratswere notcross-fostered. a. Detaileddescriptionosf allprocedures used inthe conduct ofthisstudy are providedinthe appropriatesectionsofthisreportand inAPPENDIX C (PROTOCOL AND AMENDMENT). 418-014:PAGE 1-2 A.I. Fo Generation Rats Assianed to Cross-Fostering: The Fo generationfemale ratswere observed forviabilitaytleasttwiceeach day ofthe study and forclinicaolbservationsofeffectsofthe testarticlea,bortions, premature deliverieasnd/ordeaths beforeand approximatelyone hour postdosage. Body weightswere recorded dailyduringthe dosage periodand at sacrificeF.eed consumption valueswere recordedweekly tocohabitationd,aily duringthe gestationperiodand on DLs 1,4, 7, 10 and 14. The female ratswere evaluatedfordurationofgestationl,engthof parturition, liftesrizesand pup viabiliattybirth.Pups thateitherappeared stillbomor that died beforeinitieaxlaminationofthe liftefrorviabiliwteyre examined forvital statusat birth.Matemal behaviorofthe dams was evaluateddailywhen the Pups were examined duringthe 21 -day postpartum period.Observed maternal behaviorwas recordedon DLs 1,4,7, 14 and 21. Twenty-four-hoururineand fecalsamples were collectedfrom DLs 21 to22 and blood samples were collectedon DL 22, viathe inferiovrena cava,from the matemal rats.The bloodsamples were centrifugedT.he urineand fecal samples and the serum were shippedto the Sponsor. AllFo generationratsassigned tocross-fostedngwere sacrificeodn DL 22 and a gross necropsy ofthe thoracic,abdominal and pelvicviscerawas performed. The number and distributiofnimplantationsiteswere recorded.A liversection (righltateralobe)from each dam was collectedand shipped tothe Sponsor. Rats thatdid notdelivera liftewrere sacrificeodn DG 25 and examined forgross lesions.A blood sample was collectedpriorto sacrificveiathe inferiovrena cava; samples were centrifugedand serum samples were shipped tothe Sponsor. A liversection(righltateralobe)from each dam was retainedand shipped tothe Sponsor. Dams with no survivingpups were sacrificeadfterthe lastpup was found dead, missingor presumed cannibalized. A.2. Fl Generation Pups Assigned to C ross-Fostering: Day 1 oflactatio(npostpartum)was definedas the day ofbirth.Each liftewras evaluatedforviabilitaytleasttwiceeach day ofthe postpartum period.The pups presentineach liftewrere counted once each day. Physicalsignswere recordedforthe pups once each day duringthe postpartum period.Pup body weightswere recordedon DLs 1 (birlh4), 7, 14 and 21 and atsacrifice. Pups found dead or sacdficedbecause of moribunditywere examined forgross lesionsand forthe cause ofdeath or the modbund condition.For allpups found dead on DLs 2 to4, the lungs and the liverwere preserved. Pups withgross 418-014:PAGE 1-3 lesionsfound on DLS 2 to4 were preserved and forpups found dead on DLs 5 to21, the lungs,liverand gross lesionswere preserved. On DL 4, cross-fostereFdl generationlifterwsere culledtofivemale and five female pups per liftewrh,ere possible.Pups withgross lesionswere preserved. The lungs and the livewrere preserved.The lungs and the livewrere collected from the firstten culledpups from each dosage group (irrespectiovfeliftefrr)om each dosage group on each day (controlpoolonlyon DL 1).The lungsand the livewrere retained.Remaining culledpups were sacrificeadnd discarded withoutevaluation. On DL 21, allpups were sacrificevdiadecapitatioannd examined forgross lesions.Gross lesionswere retained.Six lifterpser subset were randomly selectedforcollectioonf pooled samples (perlifteorf)bloodand liver.Blood samples were collectedviathe inferiovrena cava from each pup, pooled (per littearn)d the samples were centrifuged.The serum samples were shippedto the Sponsor. A.3. Fo Generation Rats Assigned to Pharmacokinetic Sample Collection: Viabilitaynd clinicaolbservations,body weights,feed consumption values and deliveryobservationswere performed as previouslydescribedforFo generation dams assigned tocross-fostering. Pharmacokineticsamples were collectedfrom eightand two Fo generationrats inGroups Iand 11,respectivelyo,n DL 14. Milksamples were collected approximatelytwo tosixhours postdosage on DL 14. The samples were shippedto the Sponsor. Followingcompletionofmilksample collectiofne,male ratswere sacrificeadnd a gross necropsy ofthethoracic,abdominal and pelvic viscerawas performed. The number and distributiofnimplantatiosniteswas recorded. Blood samples were collectedviathe inferiovrena cava and centrifugedand the serum was shipped tothe Sponsor. A livesrectionand the milk-secretingglands from the axillaryt,horacic,abdominal and inguinalregions ofeach dam (lefstideonly)were collectedand shippedtothe Sponsor. A.4. Fl Generation Pups Assigned to Pharmacokinetic Sample Collection: Viabilitaynd clinicaolbservationsb,ody weightsand lifteorbservationswere performed as previouslydescribedforFl generationpups assigned tocrossfosteringO.n DL 14,lifterasssignedto the pharmacokineticsample collection were sacrificeadnd blood samples were collectedviathe inferiovrena cava from each pup, pooled (perlifterc)e,ntrifugedand the serum was shipped tothe Sponsor. The liverfrom each pup was collectedp,ooled (perlittera)n,d shipped to the Sponsor. 418-014:PAGE 1-4 B. Results AllFo generationfemale ratssurvivedto scheduled sacrificeA.lladverse clinical and necropsy observationswere consideredunrelatedtothe administratioonf PFOS. Body weight gainswere reduced inthe 1.6mg/kg/day dosage group duringthe precohabitatioannd gestationperiods,as compared tothe controlgroup values. Matemal body weight gainswere variablethroughoutthe lactatiopneriodand body weight gains forthe entirelactatiopneriodwere generallycomparable among the fourcross-fosteresdubsets.Absolutebody weightswere slightly reduced inthe 1.6mg/kg/day dosage group beginningatthe end ofthe precohabitatiopneriodand continuingthroughthegestationand lactation periods. Absoluteand relativfeeed consumption valueswere generallyreduced forrats administeredthe 1.6mg/kg/day dosage ofPFOS duringthe precohabitation, gestationand lactatiopneriods,as compared tothe controlgroup values. The durationofgestationtended to be reduced inthe 1.6mg/kg/day dosage group. As observed ina previouslyconducted study(ArgusResearch LaboratoriesI,nc.,Protocol418-008),thisreductionwas associatedwithminimal preimplantatiolnoss;the average number ofimplantatiosnitesper dam was slightlryeduced,resultingina slightlryeduced deliveredliftesrize.The duration ofdeliveryper pup (inminutes)tended to be reduced inthe 1.6mg/kg/day dosage group. The liveliftesrizebeforecross-fostednginthe 1.6mg/kg/day dosage group was alsoslightlryeduced. The gestationindex was 100% ineach dosage group;allpregnantratsdeliveredliveoffspring. Pup mortalitwyas greatestinpups from testarticle-treatdeadms fosteredby test article-treatdeadms (subsetC). Pup mortalitwyas also increasedinpups from testarticle-treatdeadms fosteredby vehicle-treateddams (subsetA). As a resultofthese increasesinpup mortalityt,he viabiliitnydices(number of live pups on DL 4 preculling/numbeorf pups cross-fostereDdL1) were reduced,and numbers ofsurvivingpups per liftearnd liveliftesrizesatweighingwere reduced on DL 4 precullinignthese two subsets. Pup mortalitiynpups from vehicletreateddams fosteredby testarticle-treatdeadms (subsetD) was comparable to thatinpups from vehicle-treateddams fosteredby vehicle-treateddams (subsetB). These observationsindicatethatinuteroexposure tothe testarticle isresponsibleforpup mortalitya,nd thatpotentiaelxposure tothe testarticlveia maternalmilkdidnot adverselyaffecttheviabilitoyfthe pups. Pup body weights were reduced on DL 1 inboth subsetsofdams administeredPFOS (subsetsC and D),as compared to pup body weightsinthe subsets ofdams administered the vehicle(subsetsA and B). Potentiaelxposure to the testarticlveiamatemal milkafterDL1 reduced pup body weights,regardlessofinuteroexposure tothe 18-014:PAG E 1-5 testarticloervehicle.AfterDL 1,pup body weightswere somewhat reduced in subsetsD (pupsfrom vehicle-treateddams fosteredby testarticle-treatdeadms) and A (pups from testarticle-treatdeadms fosteredby vehicle-treatdeadms), as compared to the pups from vehicle-treateddams crossfosteredtovehicletreateddams (subsetB). AfterDL 1,pup body weightreductionswere greatest insubset C (pups from testarticle-treatdeadms fosteredby testarticle-treated dams). Two lifterisnsubset A (pups from testarticle-treatdeadms cross-fosteretdo vehicle-treateddams) and one lifteirnsubsetC (pups from testarticle-treated dams cross-fosterewdithtestarticle-treatdeadms) had pups thatdidnot nurse; pup mortalitwyas greatestinthese two subsets.There was no milkinthe stomach of 57.1%, 100% and 87.0% ofthe pups thatwere found dead and necropsiedinsubsets A, C and D, respectively. Evaluationofthe liversofpups thatdied orwere sacrificeodn DL 1 by electron microscopy revealedincreasednumbers of peroxisomes withinhepatocytesof pups from dams treatedwith 1.6mg/kg/day PFOS, compared withcontrols. Quantitatioonf peroxisomes indicatedslightlmyore than twicethe number of peroxisomes intreatedpups. Differencesincellulamrembranes or mitochondria were notdiscerniblbeetween controland treatedpups inthe samples examined. Evaluationofthe lungsof pups thatdied orwere sacrificeodn DL 1 withelectron microscopy revealedType 11pneumocytes containinglamellarbodies inboth controland 1.6mg/kg/day dosage group pups, withan increasednumber of Type 11pneumocytes and lamellarbodies inlungsfrom treatedpups. Although some lamellarmaterial(surfactantw)as identifiabwliethinalveolarlumina,no significandtifferencweas noted between the controlgroup and 1.6mg/kg/day dosage group. C. Conclusion 418-014:PAGE 1-6 The 1.6 mg/kg/day dosage of PFOS caused reductionsinbody weight gain and reduced feed consumption values forthe Fo generationdams, as occurredina previousmultigeneratiosntudy withthe testarticl(eArgus Research LaboratoriesI,nc.,Protocol418-008). As previouslyobserved,the 1.6 mg/kg/day dosage of PFO.S administeredtothe Fo generationdams inthis study also reduced the durationsofgestationand parturitiocna,used preimplantatiolnossand reductionsinliftesrizeand pup growth. Cross-fostedngof pups from testarticltereateddams and vehicletreateddams revealedthatthe effectson pup mortalittyhatoccurredinthe previously conducted study(418-008),can be attributetdoinuteroexposure tothe test articlea,nd thatpotentiaelxposure to the testarticlveiamatemal milkdid not adverselyaffectthe viabilitoyfthe pups. Exposure tothe testarticlveiaboth in uteroexposure and matemal milkcaused reductionsinpup growth. MildredS. ChristianP,h.D.,Fellow/A,TS Date ExecutiveDirectorof Research . ... ........2 3- ITL/i-) Y Alan M. Hoberman, Ph.D.,DABT Date Directorof Research @aero ondc G. York,P D., ABT Date AssociateiiDireryecco@rttoor'roopff eeParch andc Study Director li. DESCRIPTION OF TEST PROCEDURES A. Conduct of Slgdy: 418-014:PAGE 11-1 A.l. Sponso 3M Corporate Toxicology,3M Center Building220-2E-02, St.Paul,Minnesota 55144-1000 A.2. Testing Fa@iii Argus Research Laboratories,Inc.,905 Sheehy Drive,BuildingA, Horsham, Pennsylvania 19044-1297 A.3. Study Number: 418-014 A.4. Sponsor's Study Number: T-6295.13 A.5. Purpose of the Stud The purpose ofthisstudywas toevaluatesurvivalofFl generationpups followingPFOS treatmentofCrl:CD@BR VAF/Plus@ Fo generationfemale rats duringpremating,gestationand lactationF.l generationpups were cross-fostereduringthe lactatiopneriodtodifferentiabteetween effectson pups exposed tothe testarticlienuteroand pups exposed tothe testarticlveia maternalmilk.Selectedpharmacokineticsamples were collectedfrom Fo generationfemale ratsand Fl generationpups. A.6. Study Design: The requirementsofthe U.S. Food and Drug Administratio(nFDA)(')were used as the basisof studydesign. A.7. Regulato[yCompliance: The studywas conducted incompliancewithGood LaboratoryPractice(GLP) regulationosfthe U.S.Food and Drug Administratio(nFDA)(')t,he Japanese Ministroyf Healthand Welfare(MHW)(')and the European Economic Community (EEC)(4).There were no deviationsfrom the GLP regulationtshataffectedthe qualityor integritoyf the study. QualityAssurance Unitfindingsderivedfrom the 418-014:PAGE 11-2 inspectionsduringthe conduct ofthisstudy are documented and have been providedto the Study Directorand the TestingFacilitMyanagement. A.8. Ownership of the Study: The Sponsor owns the study. Allraw data,analyses,reportsand preserved tissuesare the Oropertyofthe Sponsor. A.9. Study Monitor: Marvin T. Case, D.V.M.,Ph.D. A.10. AlternateStudy Monito Andrew M. Seacat, Ph.D. A.11. Study Director: Raymond G. York,Ph.D.,DABT (AssociateDirectorofResearch) A.12. Technical Performance: John F. Barnett,B.S.(DirectorofLaboratoryOperations) MargaretM. Martin(Research Associate) CorrieL.Pabst (QualityControlAssociate) A.13. Report Preparation: Raymond G. York,Ph.D.,DABT Jo Ann Frazee,M.S. (StudyCoordinator) Susan K. Bradshaw, B.S.(Data Management Specialist) Karen G. Parker,A.A. (ReportAdministrator) A.14. Report Review: Alan M. Hoberman, Ph.D.,DABT (Directoorf Research) MildredS. ChristianP,h.D.,Fellow,ATS (ExecutiveDirectorof Research) A.15. Date Protocol Signed: 23 October 1998 A.16. Dates of Technical Performance: 418-014:PAGE 11-3 Rat Arrival 27 OCT 98 Dosage Period(42 days beforecohabitation untilDL a 21) 02 NOV 98 -28 JAN 99 CohabitationPeriod 13 DEC 98 PM - 19 DEC 98 AM Day 0 of Presumed Gestation(DG 0) DeliveryPeriod 14 DEC 98 - 19 DEC 98 04 JAN 99 -09 JAN 99 DL 1 Culling(ControlPool) 04 JAN 99 -09 JAN 99 DL 4 Culling(Cross-FosteredLftters) 08 JAN 99 - 11 JAN 99 DG 25 Sacrifice 08 JAN 99 - 12 JAN 99 DL 14 PharmacokineticGroup Sacrifice 17 JAN 99 -22 JAN 99 Fl GenerationPups Sacrific(eDL 21) 25 JAN 99 -28 JAN 99 Fo GenerationDams Sacrific(eDL 22) 26 JAN 99 -29 JAN 99 A.17. Records Maintained: The originarleport,raw data and reservesamples ofthe bulktestarticlaend vehiclecomponents are retainedinthe archivesofArgus Research Laboratories, Inc.Any preservedtissuesare retainedinthe archivesof theTestingFacilitfyor one year afterthe mailingofthedraftfinalreport,afterwhich the Sponsor will decide theirfinaldispositionA.llunused preparedtestarticlfeormulationwsere discardedattheTestingFacilityU.nused bulktestarticlweas returnedtothe Study Monitor. B. Test ArticleInformation: B.I. Description: PFOS (FC-95)-a light-colorpeodwder B.2. Lot/BatchNumber: 217 (Expiratiodnate: May 2000) B.3. Date Received and Storage Conditions: The testarticlweas receivedon 21 October 1998, and storedat room temperature. a. DL isused as an abbreviationforday of lactatioonr day postpartum. 418-014:PAGE 11-4 B.4. Special Handling Instructions: Standard safetyprecautions(use of protectiveclothing,gloves,dust-mist respiratoarnd safetygoggles)were taken when handlingthe bulktestarticlaend prepared formulations. B.5. Analysis of Activi Informationregardingthe identitys,trength,compositionand purityofthe test articlieson filweiththe Sponsor. C. Vehicle Information: C.I. Description: 0.5% Tween@ 80 inReverse Osmosis Membrane Processed DeionizedWater (R.O.DeionizedWater) C.2. Lot Numbers: Tween@ 80 - M03HO5, L06662 and M29477 C.3. Dates Received and Storage Conditions: The Tween@ 80 was receivedon 3 December 1998 (lotM03HO5), 1 September 1998 (lotL06662) and 17 September 1998 (lotM29477), from J.T.Baker, PhillipsburNge,w Jersey,and was storedatroom temperature.The R.O. deionizedwater isavailablefrom a continuoussource atthe TestingFacility and ismaintainedatroom temperature. C.4. Special Handling Instructions: Standard safetyprecautions(use of protectivcelothingg,loves,dust-mist respirators,afetygoggles or safetyglassesand a face-shieldw)ere taken when handlingthe vehicle. C.5. Analysisof Purit Neitherthe Sponsor northe Study Directorwas aware ofany potential contaminantslikeltyo be presentinthe vehiclethatwould interferweiththe resultsof thisstudy. 418-014:PAGE 11-5 D. Test ArticlePreparation and Storage C )nditions: Suspensions were prepared dailyat the Testing Facilitayt concentrationsof 0 and 0.32 mg/mL. Records are maintainedinthe raw data todocument how thetestarticlfeormulationswere prepared. Prepared suspensionswere stored at room temperature. D.I. Sample Information: Sample Type Concentration(allevels) Test ArticleReserve VehicleComponents Reserve R.O. DeionizedWater Tween(D 80 LotL06662 Lot M29477 Lot M03HO5 Size 2 mL* Ig Date Retained 01 NOV 98 27 JAN 99 28 OCT 98 5mL 28 OCT 98 5mL 28 OCT 98 5mL 10 NOV 98 5 mL 1 09 DEC 98 Shipping/Storage Conditions Shipped To Date Shipped Frozen on dry ice Sponsor 02 NOV 98 27 JAN 99 Room temperature TestingFacility 25 NOV 98 Archives Room temperature TestingFacility Archives 25 NOV 98 25 NOV 98 25 NOV 98 27 JAN 99 a. Duplicatesamples were taken from thefirsatnd lastpreparationon theday prepared. One setof samples was sentforanalysis;theremainingsamples were retainedat theTestingFacilitays backups. Backup samples were stored(-70*Cor below)and willbe discardedattheTestingFacilituypon requestoftheSponsor. D.2. AnalylicalResults: Data verifyintghe stabiliotfythe testarticlienthe vehiclefor48 hours under the conditionsofadministratioanre on filewiththe Sponsor. Homogeneity ofthe prepared formulationsison filweiththe Sponsor. Resultsofthe concentration analyses have notbeen received. E. Test System: E.l. Species: Rat E.2. Strain: CH:CDOBR VAF/Plus@ (Sprague-Dawley) E.3. Supplier (Source 418-014:PAGE 11-6 Charles River Laboratories,Inc.,Portgage, Michigan E.4. Sex: Female (Male ratsofthe same source and strainwere used onlyas breeders and were not consideredpartof theTest System). E.5. Rationale forTest System: The CD:CD@BR VAF/Plus@ (Sprague-Dawley) ratwas selectedas the Test System because: 1)thisstrainof ratwas used inthe reproductivaend developmentaltoxicitsytudies;2) historicadlata and experienceexistatthe TestingFacilit5-v7);and 3)the testarticliespharmacologicallyactiveinthe speciesand strain. E.6. Test System Data: Number ofRats Approximate Date ofBirth Approximate Age atArrival Weight (g)on the Day afterArrival Weight (g)atStudy Assignment E.7. Breeder Male Rat Data: 86 23 AUG 98 66 days 179-226 200-244 Number of Rats Approximate Date ofBirth Approximate Age atArrival Weight (g)on the Day AfterArrival Weight (g)atCohabitation E.8. Method of Randomization: 110 13 JAN 98 77 days 317-352 514-866 Upon arrivalm,ale and female ratswere assigned toindividuahlousing on the basisofcomp uter-generatedrandom units.Afteracclimation7,8 virginfemale ratswere selectedforstudy on the basisofphysicalappearance and body weightsrecordedduringacclimation.The female ratswere assigned todosage groups (42 inthe controlgroup and 36 inthe treatedgroup)based on computergenerated (weight-orderedr)andomizationprocedures. Withineach dosage group, consecutiveorderwas used to assignfemale ratsto cohabitatiownithbreeder male rats,one male ratper female rat.Rats were assigned tocross-fosterinogr pharmacokineticcollectiodnepending on the order 418-014:PAGE 11-7 of deliveryand the availabiliotfyratsforcross-fosteringE.ightfemale ratsfrom the controlgroup and two female ratsfrom the treatedgroup were designatedfor pharmacokineticsample collectiofnollowingdeliveryt;hese ratswere not crossfostered'. Twenty-fivelifterpser dosage group were cross-fostereodn DL 1b.The lifter-s were assigned such thatfoursubsetswere obtainedamong the two dosage groups,as describedinthe tablebelow. Subset assignments were made based on orderofdeliveries. Dosage Group 1 1 11 11 Number of Mated Female Rats 13 12 12 13 Dosage (mg/kg/day) 0 (Vehicle) 0 (Vehicle) 1.6 1.6 Number of Litters Reassigned 13 12 12 13 Reassigned Lifter Type Subset (Dam Treatment) Number Treated A Untreated B Treated c Untreated D On DL 4,cross-fostereldifterwsere culledtofivemale and fivefemale pups per liftewrh,ere possible,using a tableof random units. E.9. System of Identification: E.9.a.Fo Generation Rats: Rats were permanently identifieudsingMonel@ self-piercinegar tag (Oey Band and Tag Co.,Inc.,No. MSPT 20101). Male ratswere givenuniquepermanent identificatinounmbers upon assignment totheTestingFacilityb'rseeder male rat population.Female ratswere assigned temporary numbers at receiptand given unique permanent identificatinounmbers when assigned tothe study. Cage tags were marked withthe study number, permanent ratnumber, sex,testarticle identificatiaonnd dosage level. E.9.b.Fl Generation Pups: Pups were not individualliydentifiedduringlactationa;llparameters were evaluatedinterms ofthe Iifter. a. See APPENDIX D (DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILTTY), items 1 and 2. b. See APPENDIX D, item 3. F. Husbandrv: 418-014:PAGE ii-8 F.I. Research FaclillyRegistration: USDA RegistratioNno. 23-R-099 under the Animal WelfareAct,7 U.S.C.2131 etseq. F.2. Study Rooms: The study rooms were maintainedunder conditionsof positiveairflowrelativteo a hallwayand independentlysuppliedwitha minimum often changes per hour of 100% freshairthathad been passed through99.97% HEPA filtersR.oom temperatureand humiditywere monitoredconstantlythroughoutthe study. Room temperaturewas targetedat640F to 790F (180C to260C);room humidity was targetedat30% to 70%. See APPENDIX E (TEMPERATURE AND RELATIVE HUMIDITY REPORTS). F.3. Housina: Fo generationratswere individuallhyoused instainlessteel,wire-boftomed cages, except duringthe cohabitationand postpartum periods.Duringthe cohabitationpedod, each pairof ratswas housed inthe male rat'scage. Beginningno laterthan DG 20, Fo generationfemale ratswere individually housed innestingboxes, except duringthe collectioinntervaflorurineand fecal samples. Duringthiscollectioinnterva(lDLs 21 to 22),the female ratswere housed individualliynmetabolism cages. Each dam and assigned liftweerre housed ina common nestingbox duringthe postpartum period.Allcage sizes and housing conditionswere incompliance withthe Guide forthe Care and Use of LaboratoryAnimals F.4. Lighting: An automatically-controlflleudorescentlightcyclewas maintainedat 12-hours light1:2-hoursdark,witheach dark periodbeginningat 1900 hours EST. F.5. Sanitization: Cage pan linerwsere changed approximatelythreetimes each week. Cages were changed approximatelyevery otherweek. Bedding was changed as often as necessary to keep the ratsdry and clean. F.6. Feed: Ratswere givenad libituamccesstoCertifieRdodent Diet@#)5002 (PMI NutritioInnternationaSlt,.Louis,Missouri)inindividuafleeders. 11-9 F.7. Feed Analysis: Analyses were routinely performed by the feed supplier. No contaminants at levelsexceeding the maximum concentrationlimitfsorcertifiefdeed or deviationsfrom expected nutritionraelquirementswere detectedby these analyses. Copies ofthe resultsof thefeed analysesare availableinthe raw data. Neitherthe Sponsor nor the Study Directorwas aware ofany potential contaminantslikeltyo have been presentinthe feed thatwould have interfered withthe resultsofthisstudy. F.8. Water: Localwater thathad been processed by passage througha reverseosmosis membrane (R.O.water)was availableto the ratsad libitumfrom an automatic wateringaccess system and/orindividuawlater boftles.Chlorinewas added to the processed water as a bactedostat. F.9. Water Analysis: The processed water isanalyzedtwiceannuallyforpossiblechemical contamination(LancasterLaboratoriesL,ancaster,Pennsylvania)and monthly forpossiblebacteriaclontamination(AnalyticaLlaboratoriesI,nc.,Chalfont, Pennsylvania).Copies ofthe resultsof thewater analysesare availableinthe raw data. Neitherthe Sponsor nor the Study Directorwas aware ofany potential contaminantslikeltyo have been presentinthe water thatwould have interfered withthe resultsofthisstudy. F.10. Beddinci: Bed o'cobs@ was used as the nestingmaterial(Andersons'IndustriaPlroducts Groups, Maumee, Ohio). F.11. Bedding Analysis: Neitherthe Sponsor nor the Study Directorwas aware ofany potential contaminants likeltyo have been presentinthe bedding thatwould have interferewdiththe resultsofthisstudy.Analyses forpossiblecontaminationare conducted annuallyand documented inthe raw data. G. Methods: G.I. Dosage Administration: 418-014:PAGE 11-10 Dosage Group Dosage (mg/kg/day)l Concentration(mg/mL) Dosage Volume (mL/kg) 0 (Vehicle) 0 5 1.6 0.32 5 a. The test articlewas considered 100% pure for the purpose of dosage calculations. Dosage Group 1 @77 171 Rats Dosed UntilCohabitation Number of Rats Assigned Numbers 42 18901 - 18942 36 1 18943 - 18978 Rats Assigned to Pharmacokinetic Evaluation Number Assigned Numbers 0 1 18910,18911,18913, 8 18915.18926,18927, 18939,18940 2 18956,18971 Dosage Group Subset Number Dams Assigned Dam and LifteNrumbers* Lifters 18903,18907,18908,18909,18918,18920, 18976,18963,18953,18961,18946,18952, I A 18924,18929,18934.18936,18937,18941, 18972,18950,18959,18977,18958,18951, 1 18942 18965 1 B 18901,18904,18906,18912,18917,18919, 18931,18932,18938,18935,18922,18923, 18923,18922,18935,18938,18932,18931 18919,18917,18912,18906,18904,18901 11 18943,18947,18948,18957,18960,18964, 18968,18973,18970,18969,18974,18967, c 18967,18974,18969,18970,18973,18968 18964.18960,18957,18948,18947,18943 11 18976.18963.18953,18961,18946,18952. 18903,18907,18908,18909,18918,18920, D 18972,18950,18959,18977,18958,18951, 18924.18929,18934,18936,18937,18941, 1 18965 18942 1 a. Dams and theircross-fostereldifterasre listedin consecutiveorder. G.2. Rationale forDosage Selection: Dosages were selectedon the basisofa previoustwo-generationstudy conducted withthe testarticl(eArgus Research LaboratoriesI,nc., Protocol418-008). Inthisstudythe Fo generationmatemal and paternalnoobservable-effect-le(vNeOlEL) ofPFOS was 0.1 mg/kg/day (0.4mg/kg/day and higherdosages caused reductionsinbody weightgain and reduced feed consumption values). The Fo generationreproductiveNOEL was greaterthan 3.2 mg/kg/day;no effectson mating,fertiliotryestrouscyclingoccurred.The NOEL forviabiliatnyd growth inthe Fl generationoffsprinwgas 0.4 mg/kg/day (1.6mg/kg/day and higherdosages caused preimplantatiolnossand reductionsinliftesrize,pup viabilitgyr,owth and survival). 418-014:PAGE 11-11 The Fl generationmatemal and paternalNOEL ofPFOS was 0.1 mg/kg/day (0.4mg/kg/day dosage caused reductionsinbody weightgain and reduced feed consumption values).The Fl generationreproductiveNOEL was greaterthan a dosage of0.4 mg/kg/day;no effectson mating or fertiliotcycurred.The NOEL forviabilitaynd growth inthe F2 generationoffspringwas 0.1 mg/kg/day (0.4mg/kg/day dosage caused stillbirtahnsd reductionsinliftesrize,pup viabilitgyr,owth and survival). G.3. Route of Administration: Oral(gavage) G.4. RationaleforRoute ofAdministration: The oral(gavage)routewas selectedforuse because: 1)thiswas the routeof administratioinnthe developmental and reproductivetoxicologystudies;and 2) it isone ofthe possibleroutesofhuman exposure. G.5. Freguency ofAdministration: G.5.a.Fo Generation Female Rats: Female ratswere giventhe testarticloer the vehicleonce dailybeginning 42 days priorto cohabitatiotnhrough DL 21. Dosages were adjusteddailyfor body weight changes and given atapproximatelythe same time each day. G.5.b.Fl Generation Pups: Fl generationpups were not directlgyiven the testarticleb,ut may have been possiblyexposed tothe testarticldeuringmaternalgestation(inuteroexposure) or viamatemal milkduringthe lactatiopneriod. G.6. Length of Stud Approximately3 months G.7. Method of Study Performance: G.7.a.Cohabitationand Cross-FosteringProcedures: Twenty-fiveratsassigned tothe controlgroup were cohabitedone day priorto cohabitatiofnorratsassigned tothe treatedgroup and the cohabitatiopneriod consistedof a maximum of sixdays'. The cohabitationperiodforthe remaining a. See APPENDIX D, item4. 418-014:PAGE 11-12 ratsinthe controlgroup and the ratsassignedtothe treatedgroup was a maximum offivedays. During cohabitationb,oth controland PFOS-treated groups were co-housed withuntreatedmale breeders,one male ratper one female rat.Female ratswithspermatozoa observed ina smear ofthe vaginal contentsand/ora copulatoryplugobserved insituwere consideredtobe at DG 0 and assigned toindividuahlousing. Severaldams assignedtothe controlgroup thatdeliveredon the firsdtay of parturitionccurred were allowedtodeliverand retaintheirnaturalpups until needed forcross-'fosterinpgurposes. This provideda poolofcontroldams availableforimmediate cross-fosterinogf pups from PFOS-treated dams, thus preventingthese pups from nursingfrom theirPFOS-treated dams. Startingwiththe deliverieosfthe dams on the second day,allpups were removed immediately(as soon as parturitiownas completed)from their respectivedams and placed witheitheranotherdam assignedtothe control group or a PFOS-treated dam. When a dam assignedtothe controlgroup was not availablet,hen a dam from the controlpoolwas used. The liftefrom that controlpooldam was then sacrificevdiadecapitation. This cross-fosterinpgrocedure resultedinfourgroups of 12 or 13 dams/pups, i.e.C,ontrol/PFOS (SubsetA),Control/Contro(lSubsetB),PFOS/PFOS (SubsetC) and PFOS/Control (SubsetD). This procedurealloweddistinctiotnos be made between prenataland postnataleffectsofthe continuousmatemal PFOS treatment. G.7.b.Fo Generation Rats Assigned to Cross-Fostering: The Fo generationfemale ratswere observed forviabiliatty leasttwiceeach day ofthe study and forgeneralappearance atleastonce duringthe acclimation period.The ratswere alsoexamined forclinicaolbservationsof effectsofthe testarticlea,bortions,premature deliverieasnd/ordeaths beforeand approximatelyone hour postdosage. Body weightswere recordedat leastonce duringthe acclimationperiod.Body weightswere recordeddailyduringthe dosage periodand at sacrificeF.eed consumption values were recordedat leastonce duringthe acclimationperiod, weekly tocohabitationd,ailyduringthe presumed gestationperiodand on DLs 1, 4, 7,10 and 14. Feed consumption was nottabulatedafterDL 14,when itwas expected thatthe pups began toconsume matemal feed. The female ratswere continualleyvaluated(24 hoursper day) forclinical observationsduringparturitio[nperformedunder red lighctonditionsduringthe dark period(timeeach pup was observed was recorded)]d,urationofgestation (DG 0 tothe time the firsptup was delivered)l,engthof partuitio(ntimeof 418-014: PAG E i'-13 observationoflastpup minus the time ofobservationofthe firsptup dividedby N-1 pups ineach lifterl)i,ftesrizes(definedas allpups delivereda)nd pup viabiliattybirth.Pups thateitherappeared stillbomor thatdied beforeinitial examinationofthe liftefrorviabiliwteyre examined forvitasltatusat birth.The lungswere removed and immersed inwater. Pups withlungsthatsank were consideredstillbomp;ups withlungs thatfloatedwere consideredlivebomand to have died shortlyafterbirth.Matemal behaviorofthe dams was evaluateddaily when the pups were examined duringthe 21-day postpartumperiod.Observed maternalbehaviorwas recordedon DLs 1,4, 7, 14 and 21. Deviationsfrom expected maternalbehaviorwere recorded,ifand when present,on allother days duringthe postpartum period. Fo generationfemale ratswere housed individualilnymetabolism cages for collectioonf urineand fecalsamples from DLs 21 to22. Followingthe 24-hour collectioinntervals,amples were collectedintocentrifugteubes,placedon dry ice,storedfrozen(-700Cor below) and shipped tothe Sponsor foranalysis. On DL 22, bloodsamples (approximately4 mL each)were collectedfrom the matemal ratsviathe infedorvena cava and transferreidntoserum separator tubes afterremoval from metabolism,caging. The samples were spun ina refrigeratecdentrifuge.The serum was transferreidntopolypropylenetubes labeledwiththe study number, ratidentificatidoant,e ofcollections,tudyday and collectiotnimepoint.Allsamples were immediatelyfrozenon dry ice,stored frozen(-700C or below) and shipped tothe Sponsor foranalysis. G.7.c.Fl Generation Pups Assi-cinedto Cross-Fostering: Day 1 oflactatio(npostpartum)was definedas the day of birthand was alsothe firsdtay on which allpups ina liftewrere individualwleyighed. Each liftewras evaluatedforviabiliattyleasttwiceeach day ofthe postpartum pedod. Dead pups observed atthese timeswere removed from the nestingbox. The pups presentineach liftewrere counted once each day. Physicalsigns (includinggrossexternalphysicalanomalies)were recordedforthe pups once each day duringthe postpartum period.Pup body weightswere recordedon DLs 1 (birth4), 7, 14 and 21 and atsacrifice. On DL 4,cross-fostereFdl generationlifterwsere culledtofivemale and five female pups per liftewrh,ere possible. 4'.8-014:PAGE 11-14 G.7.d.Fo Generation Rats Assi-aned to Pharmacokinetic SamDle Collection: Viabilitaynd clinicaolbservations,body weights,feed consumption values and deliveryobservationswere performed,as previouslydescribedforFo generation dams assigned to cross-fostering. Blood,milkand liversamples were collectedfrom eightand two Fo generation ratsinGroups Iand 11,respectivelyd,esignatedforpharmacokineticsample collectiopner dosage group on DL 14. Milksamples were collected approximatelytwo to sixhours postdosage on DL 14. Each dam was removed from the nestingbox and individualhloyused forapproximatelyfourhours. The dam was injectedintravenouslwyithone unitofoxytocin(20 USP units/mL,Lot Number 52064, expiratiodnate July,2000, manufactured by Osborn) approximatelyfiveminutes beforemilksamples (atleast100 pL per sample) were collected.The samples were immediatelyfrozenon dry ice,storedfrozen (-700Cor below)and shipped tothe Sponsor. Followingmilksample collectionb,lood samples (approximatel4y mL each) were collectedfrom the matemal ratsviathe inferiovrena cava and transferreidnto serum separatortubes. The samples were spun ina refrigeratecdentrifuge. The serum was transferreidntopolypropylenetubes labeledwiththe study number, ratidentificatidoant,e ofcollections,tudy day and collectiotnimepoint. Allsamples were immediatelyfrozenon dry ice,storedfrozen(-700C or below) and shipped tothe Sponsor. Followingcollectioonf milkand blood samples, a liversection(rightlaterallobe) and the milk-secretingglands from the axillaryt,horacica,bdominal and inguinal regionsofeach dam (lefstideonly)were collecteds,toredfrozen(-700Cor below) and shippedto the Sponsor. G.7.e.Fl Generation Pups Assigned to Pharmacokinetic Sample Collection: Viabilitaynd clinicaolbservations,body weight and lifteorbservationswere performed as previouslydescribedforFl generaton pups assigned tocrossfostering. Caps and labeledtubes were weighed (combined weight,tothe nearest0.001 g) beforeand afterretentionof pooled pup samples forsubsequent use in pharmacokineticanalyses. Blood samples were collectedviathe inferiovrena cava from each pup on DL 14, pooled (perliftearn)d transferreidntoserum separatortubes. The samples were spun ina refrigeratecdentrifuge.The serum was transferreidnto polypropylenetubes labeledwiththe study number, pup identificatidoant,e of 418-014:PAGE 11-15 collections,tudyday and collectiotnimepoint.Allsamples were immediately frozenon dry ice,storedfrozen(-700Cor below) and shippedtothe Sponsor for analysis. The liverfrom each pup was collected,pooled (perlifters)t,oredfrozen(-700Cor below) and shippedto the Sponsor foranalysis. H. Gross Necropsya: AllFo generationratswere sacrificebdy carbon dioxideasphyxiationa,nd a gross necropsy ofthe thoracic,abdominal and pelvicviscerawas performed. Gross lesionswere retainedinneutralbuffered10% formalinforpossiblefuture evaluation.Representativephotographs of matemal gross lesionsare available inthe raw data. H.I. Fo Generation Rats: The female ratswere sacrificeodn DL 22. The number and distributionf implantatiosniteswere recorded.A liversection(righltateralobe)from each dam was collecteds,toredfrozen(-700C or below) and shipped tothe Sponsor foranalysis. Followingcompletionof milksample collectioonn DL 14,female ratsassignedto pharmacokineticcollectiownere sacrificedB.lood and livesramples were collectedas previouslydescribed.The number and distributioofnimplantation siteswas recorded. Rats thatdidnotdelivera liftewrere sacrificeodn DG 25 and examined forgross lesions.Uteriwere stainedwith 10% ammonium sulfideto confirmthe absence of implantatiosnites('). Dams withno survivingpups were sacrificeadfterthe lastpup was found dead, missingor presumed cannibalized.A blood sample (approximately4 mL) was collectedp,riorto sacrificev,iathe infedorvena cava and transferredintoserum separatortubes.The samples were spun ina refrigeratecdentrifuge.The serum was transferreidntoa polypropylenetube labeledwithstudynumber, rat identificatidoant,e of collectiosnt,udy day and collectiotnimepoint.The samples were immediatelyfrozenon dry ice,and storedfrozen(-700C or below) and a. A tableof random unitswas used to selectone Fo generationcontrol group female ratand one Fl generationcontrolgroup male and female pup from which alltissuesexamined at necropsy were retained,inorder to providecontroltissuesforpotentiaclomparative histopathological evaluations. 418-014:PAGE 11-16 shipped tothe Sponsor foranalysis.A livesrection(righltateralobe)from each dam was collecteds,toredfrozen(-700C or below) and shippedtothe Sponsor foranalysis. H.2. Fl Generation Pups: Pups thatdied beforeexaminationofthe liftefrorpup viabiliwteyre evaluatedfor vitasltatusat birtha,s previouslydescribed.Pups withgross lesionswere preservedinBouin'ssolutionforpossiblefutureevaluation.The lungswere preservedinBouin'ssolutiona,nd the livewras preservedinneutralbuffered 10% formalinforpossiblefutureevaluation. Pups from dams assigned tothe controlpooland culledon DL 1 and allother pups culledon DL 4 were sacrificevdiadecapitationT.he lungsand the liver were collectedfrom the firstten culledpups from the controlpool(irrespectivoef liftearn)d the firstten pups thatwere found dead (no autolysisf)rom each dosage group on DL 1. The lungswere retainedinBouin'ssolutiona,nd the liverwsere retainedinneutralbuffered10% formalinforpossiblefuture evaluation.Remaining culledpups were sacrificeadnd discardedwithout evaluation. Pups found dead or sacrificebdecause of moribunditywere examined forgross lesionsand forthe cause of death or the moribund condition.For allpups found dead on DLs 2 to4, the lungswere preservedinBouin'ssolutiona,nd the liver was preservedinneutralbuffered10% formalin,forpossiblefutureevaluation. Pups withgross lesionsfound on DLs 2 to4 were preservedinBouin'ssolution forpossiblefutureevaluation.For allpups found dead on DLs 5 to 21,the lungs, livearnd gross lesionswere preserved inneutralbuffered10% formalinfor possiblefutureevaluation. On DL 14, lifterassigned tothe pharmacokineticsample collectioannd their respectivedams were sacrificebdy carbon dioxideasphyxiation.Individual, pooledsamples (perlifteorf)bloodand livewrere collectedas previously described.The pups were examined forgross lesions. On DL 21,allpups were sacrificevdiadecapitationand examined forgross lesions.Gross lesionswere retainedinneutralbuffered10% formalin.Six lifters per subsetwere randomly selectedforcollectioonfpooled samples (perlifteorf) blood and liver.Blood samples were collectedviathe inferiovrena cava from each pup, pooled (perliftearn)d transferredintoserum separatortubes. The samples were spun ina refrigeratecdentrifuge.The serum was transferretdo polypropylenetubes labeledwiththe study number, ratidentificatidoant,e of collections,tudy day and collectiotnimepoint.Allsamples were immediately frozenon dry ice,storedfrozen(-700Cor below)and shipped tothe Sponsor for analysis. 418-014:PAGE 11-17 Allpups selectedforcontinuedobservationwere sacrificevdiadecapitatioannd examined forgross lesions. 1. StatisticaAlnalyses: Averages and percentages were calculated.Liftevralueswere used where appropriate. 111. RESULTS 418-014:PAGE 111-1 A. Mortaliiy. Clinical and Necrot)sy Observations and 2: Individual Data - Tables 19 and 20) (Summaries - Tables I All rats survived to scheduled sacdfice. Alladverse clinicaolbservationsdudng the precohabitationg,estationand lactatiopneriodswere consideredunrelatedto thetestarticlbeecause the incidenceswere notdosage-dependent and/orthe observationsare commonly seen inratsinthe laboratoryenvironment. These observationsincluded chromorhinorrhea,scalytaila,brasionon the head, neck,taialnd/orforelimb, missing,broken and/ormisalignedincisorsl,ocalizedalopeciaon the head, limbs,neck and/orback,chromodacryorrhea,scab on the head ortail, dehydration,perinealhernia,broken or swollenforepaw digitand protruding xiphoid. The only necropsy observationwas an inguinalmass (presumed to be a galactocelei)none controlgroup dam assignedto pharmacokineticsample collectioonn lactatiodnay 14 (DL 14). B. Body Wei-ciht(sFic3uresI and 2a Summaries -Tables 3 through 8: IndividualData - Tables 21 through 231 Body weightgainsdudng the precohabitatiopneriodwere reduced as compared to the controlgroup values,beginningon days 15 to 22 ofthe study (DSs 15 to 2) and continuingthroughDSs 29 to 36. Rats inboth dosage groups lost weight on DSs 36 to43; the body weight losswas greaterinthe 1.6mg/kg/day dosage group. Although calculatedfora seven-day intervalt,he weight losses generallyoccurredon days 42 to 43 ofthe studyand were associatedwith movement ofthe male breederratsintothe animal room and mating activitiyn the animal room when the ratswere cohabitedon day 42. As a resultofthese changes, body weightgainsforthe entireprecohabitatiopneriod(DSs 1 to43) were reduced and absolutebody weightswere slightlryeduced on DS 43 (96.6% ofcontrol)inthe 1.6mg/kg/day dosage group. Matemal body weight gains continuedtobe reduced inthe 1.6mg/kg/day dosage group inthefirsttwo weeks of thegestationpedod (days 0 to 14 of gestationD,Gs 0 to 14). As a resultb,ody weightgainswere reduced forthe entiregestationperiod(DGs 0 to20) inthe 1.6mg/kg/day dosage group. Matemal body weightstended to be reduced inthe 1.6 mg/kg/day dosage group throughoutgestation. Reduced matemal body weights persistedinthe two subsets ofrats administered1.6mg/kg/day PFOS dudng the lactatiopneriod(subsetsC and D), 418-014:PAGE 111-2 as compared tothe ratsinthe subsetsadministeredthe vehicle(subsetsA and B). Lactationbody weightshad the expected degree ofvariabilibtuytwere comparable between subsetsA and B (vehiclecontrolgroups)and between subsetsC and D (testarticle-treatgerdoups). Matemal body weightgainsforthe entirelactatiopneriod(DLs 1 to 22) were essentiallcyomparable among thefour subsets. C. Absolute (g/day)and Relative(g/kg/day)Feed Consumption Values (Summaries - Tables 9 through 14: IndividualData -Tables 24 throucih26) Absolute (g/day)and relativ(eg/kg/day)feed consumption valueswere generally reduced forratsadministeredthe 1.6mg/kg/day dosage of PFOS duringthe precohabitationg,estationand lactatiopneriods,as compared to the controlgroup values. D. Natural Delive!yand LifterObservations (Summaries -Tables 15 and 16: IndividualData -Tables 27 throuah 31) D.l. Natural Delive[y Observations The durationofgestationtended to be reduced inthe 1.6mg/kg/day dosage group, bothwhen the durationofgestationwas calculatedinwhole days and to the nearesttenthofa day. As previouslyobserved,thisreductionwas associatedwithminimalpreimplantatiolnoss;the average number of implantatiosnitesper dam was slightlryeduced, resultinigna slightlryeduced deliveredliftesrize.The durationofdeliveryper pup (inminutes)tended to be reduced inthe 1.6 mg/kg/day dosage group. The livelittesrizebeforecrossfosteringinthe 1.6mg/kg/day dosage group was alsoslightlryeduced,an observationrelatedtototalliftesrizeand associatedwithstillbirths. The gestationindexwas 100% ineach dosage group;allpregnantratsdelivered liveoffspring. D.2. Cross-Fostered LifterObservations Aftercross-fosterinogn DL 1,liveliftesrizeswere comparable among the four subsets. Pup mortalitwyas greatestinsubset C (pups from testarticle-treatdeadms fosteredby testarticle-treatdeadms); 19.2% ofthe pups were found dead or presumed cannibalizedon DLs 2 to 4. Pup mortalitwyas alsoincreasedin subsetA (pups from testarticle-treatdeadms fosteredby vehicle-treatdeadms) on DLs 2 to4; 9.0% of these pups were found dead or presumed cannibalized. As a resultofthese increasesinpup mortalityt,he viabilitiyndices(number of 418-014:PAGE III-'O livepups on DL 4 preculilng/number ofpups cross-fostereodn DL1) were reduced,and numbers ofsurvivingpups per liftearnd liveliftesrizesat weighing were reduced on DL 4 precullinignthese two subsets. Pup mortalitiynsubset D (pups from vehicle-treateddams fosteredby testarticle-treatdeadms) was comparable to thatinsubset B (pups from vehicle-treateddams fosteredby vehicle-treateddams). These observationsindicatethatinuteroexposure to the testarticliesresponsibleforpup mortalitya,nd thatpotentiaelxposure to thetest articlveiamatemal milkdid notadverselyaffectthe viabiliotfythe pups. Pup body weight/liftweerre reduced on DL 1 inboth subsetsof pups from dams administeredPFOS (subsetsA and C),as compared to pup body weights inthe subsetsof pups from dams administeredthe vehicle(subsetsB and D). Potentialexposure tothe testarticlveiamatemal milkafterDL1 reduced pup body weights,regardlessofinuteroexposure to thetestarticloer vehicle.After DL 1,pup body weightswere somewhat reduced insubsets D (pups from vehicle-treateddams fosteredby testarticle-treatdeadms) and A (pupsfrom test article-treatdeadms fosteredby vehicle-treateddams), as compared tothe pups from vehicle-treateddams fosteredby vehicle-treateddams (subsetB). After DL 1,pup body weight reductionswere greatestinsubset C (pups from test article-treatdeadms fosteredby testarticle-treatdeadms). Sex ratiosand the lactatioinndex(number oflivepups on DL 21/number of live pups on DL 4 postcullingw)ere comparable among the fourcross-fostered subsets. E. ClinicalObservations from Birthto Day 21 Postpartum and Necropsy Observations (Summaries -Tables 17 and 18: IndividualData Tables 32 and 33) Two lifterisnsubset A (pups from testarticle-treatdeadms fosteredby vehicletreateddams) and one lifteirnsubset C (pups from testarticle-treatdeadms fosteredby testarticle-treatdeadms) had pups thatdid notnurse;pup mortality was greatestinthese two subsets. There was no milkinthe stomach of 57.1%, 100% and 87.0% of the pups thatwere found dead and necropsiedinsubsetsA, C and D, respectively. Allotherclinicaalnd necropsy observationsinthe Fl generationpups were consideredunrelatedtothe testarticlbeecause: 1)the incidenceswere not dosage-dependent;or 2)the observationoccurredinonlyone lifterC.linical observationsincludedblacktipoftaill,aboredbreathingd,ark incolor,missing hindpaw digitand bent tail.One pup insubset A had slightdilatioonfthe renal pelvisatnecropsy on DL 21. 418-014:PAGE 111-4 F. Electron MicroscORic Evaluation of Liver and Lun-c3from Day I POStRartum Pups (APPENDIX G) F.l. Liver Ultrastructruallyt,here were increased numbers of peroxisomes within hepatocytesofpups from dams treatedwith1.6 mg/kg/day PFOS, compared withcontrols.Quantitationofperoxisomes indicatedslightlmyore than twicethe number ofperoxisomes intreatedpups. Differencesincellulamrembranes or mitochondda were notdiscerniblbeetween controland treatedpups inthe samples examined. F.2. L"n Type 11pneumocytes containinglamellarbodieswere identifieidnboth control and 1.6mg/kg/day dosage group pups; however,thereappeared to be an increasednumber ofType 11pneumocytes and lamellarbodies inlungsfrom treatedpups. Althoughsome lamellarmaterial(surfactantw)as identifiable withinalveolarlumina,no significandtifferencweas noted between the control group and 1.6 mg/kg/day dosage group. REFERENCES 418-014:PAGE 111-5 1. U.S. Food and Drug Administratio(n1994). InternationCaolnference on Harmonisation;Guidelineon detectionoftoxicittyo reproductionfor medicinalproducts.FederalRegister,September 22, 1994, Vol.59, No. 183. 2. U.S. Food and Drug Administration.Good LaboratoryPractice Regulations;FinalRule. 21 CFR Part58. 3. Japanese Ministryof Health and Welfare (1997). Good Laboratory Practice Standard forSafety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. 4, European Economic Community (1989). Council decisionon 28 July 1989 on the acceptance by the European Economic Community of an OECD decisionlrecommendation on compliance withprinciplesof good laboratorypractice.OfficiaJlournal of the European Communities: Legislation3.2 (No.L 315; 28 October):1-17. 5. Christian,M.S. and Voytek, P.E. (1982). In Vivo Reproductive and Mutagenicity Tests. Environmental Protection Agency, Washington, D.C. NationalTechnicalInformationService,U.S. Department of Commerce, SpringfieldV,A 22161. 6. ChristianM,.S. (1984).Reproductivetoxicitaynd teratologeyvaluationsof naltrexone(ProceedingsofNaltrexoneSymposium, New York Academy of Sciences,November 7, 1983),J.Clin.Psychiat.45(9):7-10. 7. Lang, P.L. (1988). Embryo and Fetal Developmental Toxicity(Teratology) Control Data in the Charles River Crl.,CD@BR Rat. Charles River LaboratoriesI,nc.,Wilmington,MA 01887-0630. (Data base providedby Argus Research LaboratoriesI,nc.) 8. Instituteof Laboratory Animal Resources (1996). Guide forthe Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 9. Salewski,E. (1964).F6rbemethode zum makroskopischen Nachweis von lmplantationsstellaemn Uterus der Rafte. Arch. Pathol.Exp. Pharmakol. 247:367. APPENDIX A REPORT FIGURES 450 400 350 300 250 200 DAY OF STUDY MATERNAL BODY WEIGHTS Figure 1 29 36 43 DAY OF GESTATION 14 20 MATERNAL BODY WEIGHTS RATS ASSIGNED TO CROSS-FOSTERING Figure2 380 370 360 &350 340 v 330 320 -A 310 10 DAY OF LACTATION a. Vehiclecontrolgroup daM3 crossfosteredwith 1.6mgtkg/daydosagegroup litters. b. Vehiclecontrolgroup dams cross-fosterewdithvehiclecontrolgroup I'lers. c 1.6=da day = groupdam s cros-sfosterewdith 1.6mg/kg/daydosage Imup litters. d@ 1.6 group dams cross-fosterewdith vehiclecontrolgroup liev's. 14 22 APPENDIX B REPORT TABLES PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 1 (PAGE 1): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii DOSAGE (MG/KG/DAY) 0 (VEHICLE) 1.6 --------------------------------------------------------------------------------------------------------------- PRECOHABITATION (DAY I TO 42 OF STUDY):A MAXIMUM POSSIBLE INCIDENCE 1764/ 42 1512/ 36 MORTALITY 0 0 CHROMORHINORRHEA 1/ 1 3/ 2 SCALY TAIL 0/ 0 35/ 1 ABRASION b 23/ 2 7/ 1 LOCALIZED ALOPECIA: TOTAL HEAD LIMBS NECK BACK 104/ 5 49/ 3 57/ 2 32/ 1 11/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 INCISORS: TOTAL MISALIGNED MISSING/BROKEN 16/ 3 14/ 2 2/ 1 0/ 0 0/ 0 0/ 0 CHROMODACRYORRHEA 5/ 1 0/ o HEAD: SCAB 2/ 1 0/ 0 ---------------------------------------------------------------------------------------------------------------- MAXIMUM POSSIBLE INCIDENCE = (DAYS x RATS)/NUMBER OF RATS EXAMINED PER GROUP. NIN = TOTAL NUMBER OF OBSERVATIONS/NUMBEROF RATS WITH OBSERVATION. a. Day 42 of study was the first day of cohabitation for rats assigned from Group I. b. Located on the head, neck, tail or left forelimb. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE I (PAGE 2): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) ii 1.6 -------------------------------------------------------------------------------------------------------------- PRESUMED GESTATION: a MAXIMUM POSSIBLE INCIDENCE 896/ 40 752/ 34 MORTALITY 0 0 LOCALIZED ALOPECIA: TOTAL LIMBS NECK BACK UNDERSIDE 21/ 2 13/ 1 0/ 0 8/ 1 0/ 0 SO/ 4 32/ 3 18/ 1 0/ 0 B/ 1 SCALY TAIL 0/ 0 21/ 1 INCISORS: MISALIGNED 39/ 2 0/ 0 CHROMORHINORRHEA 4/ 2 0/ 0 CHROMODACRYORRHEA 15/ 1 0/ 0 DEHYDRATION 1/ 1 0/ 0 PERINEAL HERNIA 1/ 1 0/ 0 -------------------------------------------------------------------------------------------------------------- MAXIMUM POSSIBLE INCIDENCE = (DAYS x RATS)/NUMBER OF RATS EXAMINED PER GROUP. NIN = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. a. Restricted to rats with a confirmed mating date. PROTOCOL 418-014.- ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN PATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE I (PAGE 3): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- SUBSET DOSAGE DOSAGE GROUP (MG/KG/DAY) A Ia 0 (VEHICLE) B Ib 0 (VEHICLE) c 11 c 1.6 --------------------------------------------------------------------------------------------------------------- LACTATION: MAXIMUM POSSIBLE INCIDENCE 286/ 13 264/ 12 246/ 12 MORTALITY 0 0 0 SCALY TAIL 0/ 0 0/ 0 0/ 0 RIGHT FOREPAW: THIRD DIGIT BROKEN 0/ 0 0/ 0 0/ 0 LOCALIZED ALOPECIA: LIMBS 0/ 0 0/ 0 0/ 0 PROTRUDING XIPHOID 0/ 0 IG/ 1 0/ 0 TAIL: ABRASION 0/ 0 11/ 1 0/ 0 TAIL: SCAB 0/ 0 4/ 1 0/ 0 RIGHT FOREPAW: THIRD DIGIT SWOLLEN 0/ 0 2/ 1 0/ 0 INCISORS: MISALIGNED 22/ 1 0/ 0 0/ 0 --------------------------------------------------------------------------------------------------------------- MAXIMUM POSSIBLE INCIDENCE = (DAYS x RATS)/NUMBER OF RATS EXAMINED PER GROUP. NIN = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. a. Vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. Vehicle control group dams cross-fostered with vehicle control group litters. C. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross-foatered with vehicle control group litters. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 2 (PAGE 1): NECROPSY OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS ---------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii DOSAGE (MG/KG/DAY) 0 (VEHICLE) 1.6 ---------------------------------------------------------------------------------------------------------------- RATS EXAMINED a N 41b 36 MORTALITY N 0 0 APPEARED NORMAL N 40 36 LEFT INGUINAL AREA: MASS N 1 0 ---------------------------------------------------------------------------------------------------------------- a. Refer to the individual clinical observations table (Table B15) for external observations confirmed at neci b. Excludes values for dam 18933, which was discarded without further evaluation. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NTJMBER: T-6295.13) TABLE 3 (PAGE 1): BODY WEIGHTS - PRECOHABITATION - SUMMARY - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii DOSAGE (MG/KG/DAY) 0 (VEHICLE) 1.6 --------------------------------------------------------------------------------------------------------------- RATS TESTED N 42 36 BODY WEIGHT (G) DAY 1 DAY a MEAN+S.D MEAN+S.D 231.4 + 8.6 246.5 + 10.0 231.2 + 9.2 246.8 + 10.5 DAY 15 MEAN+S.D 257.9 + 11.5 257.9 + 14.1 DAY 22 MEAN+S.D 266.3 + 14.2 26S.4 + 11.9 DAY 29 MEAN+S.D 274.7 + 14.7 270.5 + 12.7 DAY 36 MEAN+S.D 285.7 + 16.5 278.3 + 14.7 DAY 43a MEAN+S.D 283.9 + 12.2 274.4 + 15.9 [ lilb --------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY [ I = NUMBER OF VALUES AVERAGED a. Last value recorded before cohabitation. b. Excludes values for rats assigned to cohabitation on day 42 of study. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 4 (PAGE 1): BODY WEIGHT CHANGES - PRECOHABITATION - SUMMARY - Fo GENERATION FEMALE RATS ---------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 1 11 DOSAGE (MG/KG/DAY) 0 (VEHICLE) 1.6 ---------------------------------------------------------------------------------------------------------------- RATS TESTED N 42 36 BODY WEIGHT CHANGE (C) DAYS I - 8 MEAN+S.D. +15.1 + 7.0 +15.6 + 8.2 DAYS 8 - 15 MEAN+S.D. +11.4 + 6.3 +11.0 + 7.5 DAYS 15 - 22 MEAN+S.D. +8.4 + 7.7 +7.5 + 4.9 DAYS 22 - 29 MEAN+S.D. +8.3 + 4.8 +5.1 + 5.9 DAYS 29 - 36 MEAN+S.D. +11.0 + 6.1 +7.8 + 5.6 DAYS 36 - 43& MEAN+S.D. -2.6 + 7.7 -3.9 + 6.2 1 li)b DAYS 1 - 43a MEAN+S.D. +53.0 + 11.4 +43.2 + 14.3 ( 17)b ---------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF STUDY [ I = NUMBER OF VALUES AVERAGED a. Last value recorded before cohabitation. b. Excludes values for rats assigned to cohabitation on day 42 of study. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 5 (PAGE 1): MATERNAL BODY WEIGHTS - GESTATION - SUMMARY - Fa GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii DOSAGE (MG/KG/DAY) 0 (VEHICLE) 1.6 --------------------------------------------------------------------------------------------------------------- RATS TESTED N 42 36 PREGNANT N 35 27 MATERNAL BODY WEIGHT (G) DAY 0 MEAN+S.D. 295.1 + 16.9 283.9 + 14.2 DAY 7 MEAN+S.D. 326.8 + 18.9 307.0 + 16.2 DAY 14 MEAN+S.D. 359.5 + 22.0 337.8 + 17.6 DAY 20 MEAN+S.D. 442.0 + 34.0 421.8 + 25.5 -------------------------------------------------------------------------------------------7-------------------- DAY - DAY OF GESTATION PROTOCOL 418-014: OPAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 6 (PAGE 1): MATERNAL BODY WEIGHT CHANGES - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii DOSAGE (MG/KG/DAY) 0 (VEHICLE) 1.6 --------------------------------------------------------------------------------------------------------------- RATS TESTED N 42 36 PREGNANT N 35 27 MATERNAL BODY WEIGHT CHANGE (G) DAYS 0 7 MEAN+S.D. +31.7 + 6.2 +23.1 + 6.7 DAYS 7 14 MEAN+S.D. +32.7 + 7.4 +30.7 + 8.7 DAYS 14 20 MEAN+S.D. +82.5 + 20.0 +84.1 + 17.1 DAYS 0 20 MEAN+S.D. +146.9 + 24.8 +137.9 + 19.9 --------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF GESTATION PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PPOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 7 (PAGE 1): MATERNAL BODY WEIGHTS - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS ---------------------------------------------------------------------------------------------------------------- SUBSET A B c DOSAGE GROUP Ia Ib II c DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0 (VEHICLE) 1.6 ---------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING N 13 12 12 INCLUDED IN ANALYSES N lle 12 12 MATERNAL BODY WEIGHT (G) DAY I MEAN+S.D. 334.8 + 20.1 331.8 + 29.9 326.6 + 20.0 DAY 4 MEAN+S.D. 335.7 + 17.6 346.7 + 29.5 326.1 + 16.1 DAY 7 MEAN+S.D. 347.6 + 11.7 359.6 + 24.9 [ illf 334.2 + 18.2 DAY 10 MEAN+S.D. 356.3 + 15.0 365.8 + 26.8 ( illf 347.6 + 16.8 DAY 14 MEAN+S.D. 368.9 + 10.5 367.9 + 30.1 1 ii)f 350.7 + 18.3 1 ilif DAY 22 MEAN+S.D. 362.0 + 9.1 366.4 + 26.0 354.7 + 19.1 --------------------------------------------------------------------------------------------------------------- DAY = DAY OF LACTATION I I = NUMBER OF VALUES AVERAGED a. vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. Vehicle control group dams cross-fostered with vehicle control litters. c. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. e. Excludes values for dams which had litters that were not cross-fostered. f. Excludes values for dam 18974, which had no surviving pups on day 4 of lactation. g. Excludes values that were associated with interrupted water access. PROTOCOL 418-014: OPAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NU14BER: T-6295.13) TABLE 8 (PAGE 1): MATERNAL BODY WEIGHT CHANGES - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS ---------------------------------------------------------------------------------------------------------------- SUBSET A B c DOSAGE GROUP Ia Ib 11 c DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0 (VEHICLE) 1.6 --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING N 13 12 12 INCLUDED IN ANALYSES N Ile 12 lif MATERNAL BODY WEIGHT CHANGE (G) DAYS 1 - 4 MEAN+S.D. +0.9 + 14.8 +14.9 + 12.4 +3.0 + 7.3 DAYS 4 - 7 MEAN+S.D. +11.9 + 10.7 +12.9 + 13.0 +8.1 + 6.7 DAYS 7 - 10 MEAN+S.D. +8.6 + 15.3 +6.2 + 8.6 +13.4 + 9.0 DAYS 10 - 14 MEAN+S.D. +12.6 + 16.6 +2.1 + 13.7 +3.1 + 9.1 DAYS 14 - 22 MEAN+S.D. -6.9 + 13.7 -1.5 + 25.2 +4.0 + 16.7 DAYS 1 - 22 MEAN+S.D. +27.2 + 15.1 +34.7 + 17.5 +31.6 + 14.6 --------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF LACTATION a. Vehicle control group dams cross-foatered with 1.6 mg/kg/day dosage group litters. b. vehicle control group dams cross-fostered with vehicle control litters. c. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. e. Excludes values for dams which had litters that were not cross-fostered. f. Excludes values for dam 18974, which had no surviving pups on day 4 of lactation. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 9 (PAGE 1): ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - PRECOHABITATION - SUMMARY - Po GENERATION FEMAL ---------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii DOSAGE (MG/KG/DAY) 0 (VEHICLE) 1.6 -----I ----------------------------------------------------------------------------------------------------------- RATS TESTED N 42 36 FEED CONSUMPTION (G/DAY) DAYS 1 8 MEAN+S.D. 19.5 + 1.7 19.7 + 2.0 DAYS 8 15 MEAN+S.D. [ 40]a 20.1 + 1.7 ( 34]a 19.7 + 1.6 ( 41]a DAYS 15 22 MEAN+S.D. 19.7 + 1.8 18.9 + 1.3 DAYS 22 29 MEAN+S.D. t 41]a 19.6 + 2.2 [ 3i)a 18.7 + 1.3 DAYS 29 - 36 MEAN+S.D. ( 4@la 19.6 + 2.1 t 341& 18.3 + 1.6 DAYS 36 - 43b MEAN+S.D. [ 40]a 18.3 + 2.4 ( 3i]a 17.2 + 1.5 DAYS I - 43b MEAN+S.D. [ 15)a,c 19.8 + 1.6 [ 3i]a 18.7 + 1.3 I lila,c ( 33)a ---------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF STUDY ( ) = NUMBER OF VALUES AVERAGED a. Excludes values that were incorrertly recorded or not recorded, as well as those associated with spillage. b. Last value recorded before cohabitation. c. Excludes values for rats assigned to cohabitation on day 42 of study. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 10 (PAGE 1): RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - PRECOHABITATION - SUMMARY - Fo GENERATION P&M ----------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii DOSAGE (MG/KG/DAY) 0 (VEHICLE) 1.6 ----------------------------------------------------------------------------------------------------------------- RATS TESTED N 42 36 FEED CONSUMPTION (G/KG/DAY) DAYS 1 - 8 DAYS 8 - 15 MEAN+S.D. MEAN+S.D. 81.6 + 6.1 ( 40)a 79.7 + 5.4 82.4 + 8.1 I 34)a 78.0 + 5.2 DAYS 15 - 22 MEAN+S.D. [ 41)a 75.0 + 5.1 72.1 + 4.6 DAYS 22 - 29 DAYS 29 - 36 DAYS 36 - 43b DAYS I - 43b MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. ( 41)a 72.7 + 6.9 ( 40)a 70.0 + 5.8 t 40]a 63.8 + 7.7 ( 15]a,c 74.2 + 4.9 t 34)a 69.7 + 3.9 [ 341& 66.8 + 4.6 [ 34)a 61.9 + 3.9 [ 3i)a 71.7 + 3.8 15)a,c 33]a ----------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF STUDY I I = NUMBER OF VALUES AVERAGED a. Excludes values that were incorrectly recorded or not recorded, as well as those associated with spillage. b. Last value recorded before cohabitation. c. Excludes values for rats assigned to cohabitation on day 42 of study. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR-S STUDY NUMBER: T-6295-13) TABLE 11 (PAGE 1): MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - GESTATION - SUMMARY - Fo GENERATION FE ---------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii DOSAGE (MG/KG/DAY) 0 (VEHICLE) 1.6 ---------------------------------------------------------------------------------------------------------------- RATS TESTED N 42 36 PREGNANT N 35 27 MATERNAL FEED CONSUMPTION (G/DAY) DAYS 0 7 DAYS 7 14 MEAN+S.D. MEAN+S.D. 24.2 + 2.7 [ 33]a 26.3 + 3.0 21.1 + 1.5 23.7 + 2.0 DAYS 14 20 MEAN+S.D. 24.8 + 2.6 25.4 + 2.5 DAYS 0 20 MEAN+S.D. 25.1 + 2.2 23.3 + 1.5 ---------------------------------------------------------------------7--------------------------7---------------- DAYS = DAYS OF GESTATION ( I - NUMBER OF VALUES AVERAGED a. Excludes values that were associated with spillage. PROTOCOI, 418-014: ORAI, (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 12 (PAGE 1): MATERNAL RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - GESTATION - SUMMARY - Fo GENEPATI -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii DOSAGE (MG/KG/DAY) 0 (VEHICLE) 1.6 -------------------------------------------------------------------------------------------------------------- PATS TESTED N 42 36 PREGNANT N 35 27 MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 0 - 7 DAYS 7 - 14 MEAN+S.D. MEAN+S.D. 76.9 + 5.8 1 33]a 76.7 + 7.8 71.2 + 4.2 73.8 + 4.7 DAYS 14 - 20 MEAN+S.D. 62.8 + 4.6 67.6 + 6.3 DAYS 0 - 20 MEAN+S.D. 72.0 + 4.1 70.7 + 3.4 -------------------------------------------------------------------------------------------7------------------- DAYS = DAYS OF GESTATION ( I = NUMBER OF VAIUES AVERAGED a. Excludes values were associated with spillage. PROTOCOL 418-014: OPAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 13 (PAGE 1): MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - LACTATION - SUMMARY - Fo GENERATION FE ---------------------------------------------------------------------------------------------------------------- SUBSET A B c DOSAGE GROUP Ia Ib II c DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0 (VEHICLE) 1.6 ---------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING N 13 12 12 INCLUDED IN ANALYSES N Ile 12 lif MATERNAL FEED CONSUMPTION (G/DAY) DAYS 1 4 MEAN+S.D. 19.1 + 4.0 24.9 + 3.6 18.8 + 3.3 DAYS 4 7 MEAN+S.D. 38.8 + 2.8 42.9 + 3.7 34.5 + 6.4 DAYS 7 10 DAYS 10 14h MEAN+S.D. MEAN+S.D. 45.0 + 3.3 1 10)g 57.$@,1, 3.9 51.3 + 4.6 [ lllg 58.2 + 4.5 41.2 + 3.6 52.0@t,@ 5.8 DAYS I 14h MEAN+S.D. 41.4 + 2.5 45.4 + 3.3 38.0 + 4.4 10]g ---------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF LACTATION ( I = NUMBER OF VALUES AVERAGED a. vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. Vehicle control group dams crosa-fostered with vehicle control litters. C. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. e. Excludes values for dams which had litters that were not cross-fostered. f. Excludes values for dam 18974, which had no surviving pups on day 4 of lactation. g. Excludes values that were associated with interrupted water access. h. Because it is presumed that the pups begin to consume maternal feed after day 14 of lactation, maternal fee were not tabulated on days 14 to 21 of lactation. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PPOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 14 (PAGE 1): MATERNAL RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - LACTATION - SUMMARY - Fo GENERATIO --------------------------------------------------------------------------------------------------------------- SUBSET A B c DOSAGE GROUP Ia Ib II c DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0 (VEHICLE) 1.6 --------------------------------------------------------------------------------------------------------------- PATS ASSIGNED TO CROSS-FOSTERING N 13 12 12 INCLUDED IN ANALYSES N lle 12 lif MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 1 4 MEAN+S.D. 57.5 + 12.5 73.8 + 11.6 58.4 + 11.1 DAYS 4 7 MEAN+S.D. 113.2 + 9.3 122.0 + 15.1 104.4 + 18.6 DAYS 7 10 DAYS 10 14h MEAN+S.D. MEAN+S.D. 127.2 + 10.4 [ 10]g 159.7 + 12.0 141.9 + 14.0 [ lljg 159.9 + 10.8 121.2 + 12.9 150.7 + 15.8 DAYS 1 14h MEAN+S.D. 118.8 + 8.9 128.5 + 11.4 [ 10]g 113.8 + 12.6 [ lolg --------------------------------------------------------------------------------------------------------------- DAYS - DAYS OF LACTATION I I = NUMBER OF VALUES AVERAGED a. vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. vehicle control group dams cross-fostered with vehicle control litters. C. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. e. Excludes values for dams which had litters that were not cross-fostered. f. Excludes values for dam 18974, which had no surviving pups on day 4 of lactation. g. Excludes values that were associated with interrupted water access. h. Because it is presumed that the pups begin to consume maternal feed after day 14 of lactation, maternal fe were not tabulated on days 14 to 21 of lactation. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PPOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 15 (PAGE 1): NATURAL DELIVERY OBSERVATIONS - SUMMARY - Po GENERATION FEMALE RATS ---------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii DOSAGE (MG/KG/DAY) 0 (VEHICLE) 1.6 ---------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING N 25 25 PREGNANT N(t) 25(100.0) 25(100.0) DELIVERED LITTERS N(%) 25(100.0) 25(100.0) INCLUDED IN ANALYSES N 23a 23a DURATION OF GESTATION IN DAYS b MEAN+S.D. 22.0 + 0.5 21.6 + 0.1 NUMBER DAYS DAYS OF DAMS DELIVERING 21.1 22.0 22.1 23.0 N(%) N(t) 14( 60.9) 9( 39.1) 23(100.0) 0( 0.0) DURATION IN WHOLE OF GESTATION DAYS c MEAN+S.D. 22.4 + 0.5 22.0 + 0.0 NUMBER OF DAMS DELIVERING DAY 22 DAY 23 N(%) N(%) 14( 60.9) 9( 39.1) 23(loo.0) 0( 0.0) GESTATION INDEX d t 100.0 100.0 NIN 23/ 23 23/ 23 ---------------------------------------------------------------------------------------------------------------- DAY(S) = DAY(S) OF GESTATION a. Excludes values for dams which had litters that were not cross-fostered. b. Calculated as the time (to the nearest tenth of a day) elapsed between confirmed mating (arbitrarily define the time the first pup was delivered. c. Calculated as the time (in days) elapsed between confirmed mating (arbitrarily defined as day 0) and the ti the first pup was delivered. d. Number of rats with live offspring/number of pregnant rats. PROTOCOL 418-014: OPAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 15 (PAGE 2): NATURAL DELIVERY OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii DOSAGE (MG/KG/DAY) 0 (VEHICLE) 1.6 -------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING N 25 25 PREGNANT N(t) 25(100.0) 25(100.0) DELIVERED LITTERS N(t) 25(100.0) 25(100.0) INCLUDED IN ANALYSES N 23a 23a OBSERVED DURATION OF DELIVERY PER PUP PER LITTER (MINUTES) TOTAL DELIVERY (MINUTES) b PERIOD OBSERVED MEAN+S.D. TOTAL OR PARTIAL PERIOD OBSERVED (MINUTES) b DELIVERY MEAN+S.D. IMPLANTATION SITES PER DELIVERED LITTER N MEAN+S.D. DELIVERED LITTER SIZE MEAN+S.D. DAMS WITH STILLBORN PUPS N(t) 11.6 + 4.8 22) 11.6 + 4.8 407 17.7 + 1.8 16.4 + 1.8 3( 13.0) 9.1 + 3.1 9.1 + 3.1 368 16.0 + 1.9 15.0 + 1.8 1( 4.3) PUPS FOUND DEAD (PRECROSS-FOSTERING) LIVE LITTER SIZE c NIN(96) MEAN+S.D. 1/373( 0.3) 16.2 + 1.8 1/344( 0.3) 14.9 + 1.8 FOSTER LITTER SIZE MEAN+S.D. 15.2 + 1.9 15.6 + 1.9 --------------------------------------------------------------------------------------------------------------- ( I = NUMBER OF VALUES AVERAGED a. Excludes values for dams which had litters that were not cross-fostered. b. Calculated as the time (in minutes) elapsed between delivery of the first and last pups, divided by n-1 pu c. Excludes values for dams with all pups dying prior to cross-fostering. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 15 (PAGE 3): NATURAL DELIVERY OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- SUBSET A B c DOSAGE GROUP Ia Ib II c DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0 (VEHICLE) 1.6 --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING N 13 12 12 PREGNANT N(t) 13(100.0) 12(100.0) 12(100.0) DELIVERED LITTERS N(t) 13(100.0) 12(100.0) 12(100.0) INCLUDED IN ANALYSES N lle 12 12 LIVE LITTER SIZE e MEAN+S.D. 16.4 + 1.6 15.9 + 2.1 14.8 + 1.9 FOSTER LITTER SIZE MEAN+S.D. 15.1 + 1.7 15.9 + 2.1 14.8 + 1.9 DAMS WITH ALL PUP DYING DAYS 1-4 POSTPARTUM (AFTER CROSS-FOSTERING) N(l) 0( 0.0) 0( 0.0) 1( 8.3) DAMS WITH ALL PUPS DYING DAYS 5-21 POSTPARTUM N(t) 0( 0.0) o( o.0) 0( 0.0) --------------------------------------------------------------------------------------------------------------- a. Vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. vehicle control group dams cross-fostered with vehicle control litters. C. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. e. Excludes values for dams with all pups dying prior to cross-fostering. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 16 (PAGE 1): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - Fl GENERATION LITTERS --------------------------------------------------------------------------------------------------------------- SUBSET DOSAGE DOSAGE GROUP (MG/KG/DAY) A Ia 0 (VEHICLE) B Ib 0 (VEHICLE) c II c 1.6 --------------------------------------------------------------------------------------------------------------- LITTERS ASSIGNED TO CROSS-FOSTERING N 13 12 12 INCLUDED IN ANALYSES N Ile 12 12 PUPS CROSS-FOSTERED N MEAN+S.D. 166 15.1 + 1.7 191 15.9 + 2.1 177 14.8 + 1.9 PUPS FOUND DEAD OR PRESUMED CANNIBALIZED DAY 1 DAYS 2- 4 DAYS 5- 7 DAYS 8-14 DAYS 15-21 NIN(T) NIN(vi) NIN(%) NIN(%) NIN(%) 0/166( 15/166( 1/109( 0/108( 0/108( 0.0) 9.0) 0-9) 0-0) 0.0) 0/191( 3/191( 0/120( 0/120( 0/120( 0-0) 1.6) 0.0) 0.0) 0.0) 0/177( 34/177( 0/107( 0/107( 0/107( 0.0) 19.2) 0.0) 0.0) 0.0) VIABILITY INDEX f % NIN 91.0 151/166 98.4 188/191 80.8 143/177 LACTATION INDEX g & 99.1 100.0 100.0 NIN 108/109 120/120 107/107 --------------------------------------------------------------------------------------------------------------- DAY(S) = DAY(S) POSTPARTUM a. Vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. vehicle control group dams cross-fostered with vehicle control litters. C. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. e. Excludes values for dams which had litters that were not cross-fostered. f. Number of live pups on day 4 (preculling) poatpartum/number of pups cross-fostered on day I postpartum. g. Number of live pups on day 21 postpartum/number of live pups on day 4 (postculling) postpartum. PROTOCOL 416-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PPOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 16 (PAGE 2): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - Fl GENERATION LITTERS ------------------------------------------------------------------------------------------------------------------ SUBSET A B c DOSAGE GROUP Ia Ib II c DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0 (VEHICLE) 1.6 ------------------------------------------------------------------------------------------------------------------ LITTERS ASSIGNED TO CROSS-FOSTERING N 13 12 12 INCLUDED IN ANALYSES N lle 12 12 SURVIVING PUPS/LITTER f DAY 1 DAY 4 PRECULLING DAY 4 POSTCULLING MEAN+S.D. MEAN+S.D. MEAN+S.D. 15.1 + 1.7 13.7 + 2.3 9.9 + 0.3 15.9 + 2.1 15.7 + 1.8 10.0 + 0.0 14.8 + 1.9 11.9 + 4.6 8.9 + 2.9 DAY 7 MEAN+S.D. 9.8 + 0.6 10.0 + 0.0 8.9 + 2.9 DAY 14 MEM+S. D. 9.8 + 0.6 10.0 + 0.0 8.9 + 2.9 DAY 21 MEAN+S.D. 9.8 + 0.6 10.0 + 0.0 8.9 + 2.9 -------------------------------------------------------------------------------------------:----------------------- DAY - DAY POSTPARTUM [ I - NUMBER OF VALUES AVERAGED a. vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. Vehicle control group dams cross-fostered with vehicle control litters. C. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. e. Excludes values for dams which had litters that were not cross-fostered. f. Average number of live pups per litter, including litters with no surviving pups. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 16 (PAGE 3): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - Fl GENERATION LITTERS -------------------------------------------------------------------------------------------------------------- SUBSET DOSAGE DOSAGE GROUP (MG/KG/DAY) A Ia 0 (VEHICLE) a Ib 0 (VEHICLE) c II c 1.6 -------------------------------------------------------------------------------------------------------------- LITTERS ASSIGNED TO CROSS-FOSTERING N 13 12 12 INCLUDED IN ANALYSES N lle 12 12 PERCENT MALE PUPS PER NUMBER OF PUPS SEXED DAY 1 MEAN+S.D. DAY 4 PRECULLING MEAN+S.D. DAY 4 POSTCULLING MEAN+S.D. DAY 7 MEAN+S.D. DAY 14 MEAN+S.D. DAY 21 MEAN+S.D. LIVE LITTER SIZE AT WEIGHING 46.9 + 9.7 46.7 + 11.0 48.7 + 6.4 49.3 + 7.4 49.3 + 7.4 49.3 + 7.4 48.4 + 14.4 49.1 + 14.3 52.5 + 8.7 52.5 + 8.7 52.5 + 8.7 52.5 + 8.7 48.8 + 16.6 49.4 + 21.5 1 illf 46.0 + 16.9 ( illf 46.0 + 16.9 f ilif 46.0 + 16.9 ( ly1f 46.0 + 16.9 ilif DAY 1 MEAN+S.D. 15.1 + 1.7 15.9 + 2.1 14.8 + 1.9 DAY 4 PRECULLING MEAN+S.D. 13.7 + 2.3 15.7 + 1.8 13.0 + 2.9 DAY 4 POSTCULLING MEAN+S.D. 9.9 + 0.3 10.0 + 0.0 1 ll)f 9.7 + 0.6 DAY 7 MEAN+S.D. 9.8 + 0.6 10.0 + 0.0 [ ilif 9.7 + 0.6 DAY 14 MEAN+S.D. 9.8 + 0.6 10.0 + 0.0 [ illf 9.7 + 0.6 DAY 21 MEAN+S.D. 9.8 + 0.6 10.0 + 0.0 f ilif 9.7 + 0.6 ( ilif ----------------------------------------------------------------------------------------------------------- DAY = DAY POSTPARTUM ( I - NUMBER OF VALUES AVERAGED a. Vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. vehicle control group dams cross-fostered with vehicle control litters. c. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. e. Excludes values for dams which had litters that were not cross-fostered. f. Excludes values for dam 18974, which had no surviving pups on day 4 postpartum. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 16 (PAGE 4): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - Fl GENERATION LITTERS --------------------------------------------------------------------------------------------------------------- SUBSET A a c DOSAGE DOSAGE GROUP (MG/KG/DAY) Ia 0 (VEHICLE) Ib 0 (VEHICLE) II c 1.6 --------------------------------------------------------------------------------------------------------------- LITTERS ASSIGNED TO CROSS-FOSTERING N 13 12 12 INCLUDED IN ANALYSES N lle 12 12 PUP WEIGHT/LITTER (GRAMS) DAY lg MEAN+S.D. 5.6 + 0.3 6.2 + 0.5 5.7 + 0.2 DAY 4 PRECULLING MEAN+S.D. 7.1 + 0.5 8.3 + 0.8 7.2 + 0.8 DAY 4 POSTCULLING MEAN+S.D. 7.3 + O.S 8.5 + 0.8 ( lljf 7.3 + 0.8 DAY 7 MEAN+S.D. 12.3 + 0.9 14.1 + 0.9 ( ilif 11.6 + 1.3 DAY 14 MEAN+S.D. 26.7 + 1.7 29.0 + 1.5 t ilif 24.6 + 2.4 DAY 21 MEAN+S.D. 41.9 + 1.8 46.3 + 2.5 t ll)f 38.9 + 3.8 ( illf --------------------------------------------------------------------------------------------------------------- DAY = DAY POSTPARTUM [ I = NUMBER OF VALUES AVERAGED a. vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. Vehicle control group dams cross-fostered with vehicle control litters. C. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. e. Excludes values for dams which had litters that were not cross-fostered. f. Excludes values for dam 18974, which had no surviving pups on day 4 postpartum. g. Day 1 pup weights for Subsets A and C reflect pups of test article treated dams. Day 1 pup weights for Su reflect pups of vehicle treated dams. The effects of cross-fostering are not reflected in these pup body PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 17 (PAGE 1): CLINICAL OBSERVATIONS FROM BIRTH TO DAY 21 POSTPARTUM - SUM14ARY - Fl GENERATION PUPS --------------------------------------------------------------------------------------------------------------- SUBSET A B c DOSAGE GROUP Ia Ib 11 c DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0 (VEHICLE) 1.6 LITTERS EXAMINED (N) 13 12 12 --------------------------------------------------------------------------------------------------------------- TRANSIENT CLINICAL OBSERVATIONS: e TOTAL FREQUENCY (DAYS x PUPS)/LITTERS WITH OBSERVATIONS TIP OF TAIL, BLACK NIN 0/ 0 15/ 1 0/ 0 LABORED BREATHING NIN 0/ 0 0/ 0 0/ 0 DARK IN COLOR NIN 0/ 0 0/ 0 0/ 0 NOT NURSING NIN 2/ 2 0/ 0 2/ 1 PERSISTENT CLINICAL OBSERVATIONS: e TOTAL FREQUENCY (DAYS x PUPS)/LITTERS WITH OBSERVATIONS RIGHT HINDPAW, SECOND DIGIT MISSING NIN 0/ 0 0/ 0 21/ 1 TAIL, BENT NIN 0/ a ii/ 1 0/ 0 --------------------------------------------------------------------------------------------------------------- NIN - TOTAL FREQUENCY (DAYS x PUPS)/LITTERSWITH OBSERVATIONS. a. Vehicle control group dame cross-fostered with 1.6 mg/kg/day dosage group litters. b. vehicle control group dams cross-fostered with vehicle control litters. C. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. e. Tabulation restricted to adverse observations; all other pups appeared normal. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 18 (PACE 1): NECROPSY OBSERVATIONS - SUMMARY - Fl GENERATION PUPS --------------------------------------------------------------------------------------------------------------- SUBSET A B c MATERNAL DOSAGE GROUP Ia Ib 11 c MATERNAL DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0 (VEHICLE) 1.6 -------------------------------------------------------------------------------------------------------------- LITTERS EXAMINED N 13 12 12 PUPS FOUND DEAD e,f (AFTER CROSS-FOSTERING) N 7 0 18 NO MILK IN STOMACH g N(t) 4( 57.1) 0( 0.0) 18(loo.0) --------------------------------------------------------------------------------------------------------------- PUPS SACRIFICED AND NECROPSIED ON 4 OR 21 POSTPARTUM f LITTERS EXAMINED N 9 8 7 PUPS EVALUATED N 79 75 73 APPEARED NORMAL LITTER INCIDENCE PUP INCIDENCE N(t) N(%) 8( 88.9) 78( 98.7) 8(100.0) 75(100.0) 7(100.0) 73(100.0) KIDNEYS: RIGHT, PELVIS, SLIGHT DILATION LITTER INCIDENCE N(%) 1( 11.1) 0( 0.0) 0( 0.0) PUP INCIDENCE N(t) 1( 1.3) 0( 0-0) 0( 0-0) --------------------------------------------------------------------------------------------------------------- a. Vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. Vehicle control group dams crosa-foatered with vehicle control litters. C. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. e. Restricted to the pups in which complete necropsies were performed. Complete necropsies were not performe autolysis or cannibalization precluded evaluation. f. Refer to the individual pup clinical observation table (Table B32) for external clinical observations con g. Analysis restricted to pups found dead and necropsied. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 19 (PACE 1): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- RAT # DESCRIPTION --------------------------------------------------------------------------------------------------------------- 18901 NO ADVERSE FINDINGS 18902 NO ADVERSE FINDINGS 16903 NO ADVERSE FINDINGS 18904 DG( 14 CHROMORHINORRHEA 18905 NO ADVERSE FINDINGS 18906 DS( 11- 17) LOCALIZED ALOPECIA: HEAD DS( le ALOPECIA NO LONGER APPARENT 18907 NO ADVERSE FINDINGS 18908 NO ADVERSE FINDINGS 18909 NO ADVERSE FINDINGS 18910 NO ADVERSE FINDINGS 18911 DS( 12- 20) INCISORS: MISALIGNED DS( 21 ) INCISORS: REALIGNED DG( 0- 17) INCISORS; MISALIGNED 18912 NO ADVERSE FINDINGS 18913 NO ADVERSE FINDINGS 18914 NO ADVERSE FINDINGS 18915 NO ADVERSE FINDINGS 18916 NO ADVERSE FINDINGS 18917 DS( 11- 19) NECK: ABRASION (0.5 CM IN DIAMETER) DS( 20 ) ABRASION HEALED 18918 DG( 7- 14) LOCALIZED ALOPECIA: BACK DG( is ) ALOPECIA NO LONGER APPARENT 18919 DS( 8- 22) LOCALIZED ALOPECIA: HEAD DS( 23 ) ALOPECIA NO LONGER APPARENT 18920 DS( 10- 44) LOCALIZED ALOPECIA: LIMBS DG( 0- 12) LOCALIZED ALOPECIA: LIMBS DG( 13 ALOPECIA NO LONGER APPARENT 18921 NO ADVERSE FINDINGS 18922 DL( 8- 18) TAIL: ABRASION (1.0 CM IN LENGTH) DL( 19- 22) TAIL: SCAB (0.5 CM X 0.3 CM) DL( 21- 22) RIGHT FOREPAW: THIRD DIGIT SWOLLEN a 18923 NO ADVERSE FINDINGS 18924 NO ADVERSE FINDINGS 18925 DS( 29- 39) LOCALIZED ALOPECIA: BACK 18926 DG( 21 ) PERINEAL HERNIA DL( 1- 14) PERINEAL HERNIA 18927 NO ADVERSE FINDINGS --------------------------------------------------------------------------------------------------------------- DS . DAY OF STUDY DG - DAY OF PRESUMED GESTATION DL - DAY OF LACTATION a. observation confirmed at necropsy. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 19 (PAGE 2): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Po GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- RAT # DESCRIPTION --------------------------------------------------------------------------------------------------------------- 18928 18929 18930 18931 18932 18933 18934 18935 18936 DS( 20 DS( 41- 42) DS( 43 DS( 5- 14) DS( 5- 31) DS( 11- 43) DS( 15- 16) DS( 17 ) DS( 19- 43) DG( 0) DS( 38- 43) DS( 38- 43) DS( 39- 43) DG( 0) DG( 0- 11) DG( 0- 20) NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMORHINORRHEA NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS INCISORS: MISSING/BROKEN INCISORS: GROWN IN NO ADVERSE FINDINGS HEAD: ABRASION (0-5 CM IN DIAMETER) LOCALIZED ALOPECIA: HEAD LOCALIZED ALOPECIA: NECK HEAD: SCAB (0.3 CM IN DIAMETER) SCAB HEALED LOCALIZED ALOPECIA: LIMBS ALOPECIA NO LONGER APPARENT CHROMODACRYORRHEA INCISORS: MISALIGNED TAIL: ABRASION (0.3 CM FROM TIP) ABRASION HEALED CHROMODACRYORRHEA INCISORS: MISALIGNED DG( 2 DG( 6 CHROMORHINORRHEA DEHYDRATION DG( 13- 15) CHROMODACRYORRHEA DG( 17- 18) CHROMORHINORRHEA DL( I- 22) INCISORS: MISALIGNED a 18937 NO ADVERSE FINDINGS 18938 DL( 7- 22) PROTRUDING XIPHOID a 18939 NO ADVERSE FINDINGS 18940 NO ADVERSE FINDINGS 18941 NO ADVERSE FINDINGS 18942 NO ADVERSE FINDINGS --------------------------------------------------------------------------------------------------------------- DS = DAY OF STUDY DG - DAY OF PRESUMED DL - DAY OF LACTATION a. Observation confirmed at necropsy. GESTATION PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 19 (PAGE 3): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 11 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- PAT # DESCRIPTION --------------------------------------------------------------------------------------------------------------- 18943 DS( 36- 42) LEFT FORELIMB: ABRASION (0.4 CM IN DIAMETER) DS( 43 ) ABRASION HEALED DS( 43- 45) LOCALIZED ALOPECIA: LIMBS DG( 0- 4) LOCALIZED ALOPECIA: LIMBS DG( 5 ALOPECIA NO LONGER APPARENT 18944 NO ADVERSE FINDINGS 18945 NO ADVERSE FINDINGS 18946 18947 NO ADVERSE NO ADVERSE FINDINGS FINDINGS 18948 18949 NO ADVERSE NO ADVERSE FINDINGS FINDINGS 18950 18951 NO ADVERSE NO ADVERSE FINDINGS FINDINGS 18952 DS( 17 CHROMORHINORRHEA 18953 NO ADVERSE FINDINGS 18954 NO ADVERSE FINDINGS 18955 NO ADVERSE FINDINGS 18956 NO ADVERSE FINDINGS 18957 NO ADVERSE FINDINGS 18958 DL( I- 22) DL( 16- 22) RIGHT FOREPAW: FOURTH DIGIT BROKEN a LOCALIZED ALOPECIA: LIMBS a 18959 18960 DG( 10- 17) NO ADVERSE FINDINGS LOCALIZED ALOPECIA; UNDERSIDE 18961 18962 18963 DS( 8- 44) DG( 0- 20) DL( I- 22) NO ADVERSE NO ADVERSE SCALY TAIL SCALY TAIL SCALY TAIL FINDINGS FINDINGS a 18964 NO ADVERSE FINDINGS 18965 NO ADVERSE FINDINGS 16966 NO ADVERSE FINDINGS 18967 NO ADVERSE FINDINGS 18968 NO ADVERSE FINDINGS 18969 NO ADVERSE FINDINGS 18970 NO ADVERSE FINDINGS --------------------------------------------------------------------------------------------------------------- DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL - DAY OF LACTATION a. Observation confirmed at necropsy. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 19 (PACE 4): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE PATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 11 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- RAT # DESCRIPTION --------------------------------------------------------------------------------------------------------------- 18971 18972 18973 18974 DS( 27- 28) DG( 5- 22) DG( 15- 22) DL( I- 2) DL( 3) DL( I- 2) DL( 3 CHROMORHINORRHEA LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: ALOPECIA NO LONGER LOCALIZED ALOPECIA: ALOPECIA NO LONGER NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NECK LIMBS LIMBS APPARENT NECK APPARENT 18975 DG( 7- 25) LOCALIZED ALOPECIA: LIMBS 18976 NO ADVERSE FINDINGS 18977 DL( I- 3) PROTRUDING XIPHOID a 18978 NO ADVERSE FINDINGS --------------------------------------------------------------------------------------------------------------- DS - DAY OF STUDY DL - DAY OF LACTATION DG - DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 20 (PAGE 1): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP PAT DAY OF PREGNANCY DOSAGES DOSAGE (MG/KG/DAY) NUMBER NECROPSY STATUS ADMINISTERED OBSERVATIONS a --------------------------------------------------------------------------------------------------------------- I 0 (VEHICLE) 18901 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18902 1 DG 25 NP 69 ALL TISSUES APPEARED NORMAL. 18903 DL 22 p 86 ALL TISSUES APPEARED NORMAL. 18904 DL 22 p 85 ALL TISSUES APPEARED NORMAL. 18905 DS 72 NP 71 ALL TISSUES APPEARED NORMAL. 18906 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18907 DL 22 p 86 ALL TISSUES APPEARED NORMAL. 18908 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18909 DL 22 p 85 ALL TISSUES APPEARED NORMAL. 18910 DL 14 p 76 ALL TISSUES APPEARED NORMAL. 18911 DL 14 p 80 ALL TISSUES APPEARED NORMAL. 18912 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18913 DL 14 p 80 ALL TISSUES APPEARED NORMAL. 18914 DG 25 NP 68 ALL TISSUES APPEARED NORMAL. 18915 DL 14 p 76 ALL TISSUES APPEARED NORMAL. 18916 DG 2S p 70 ALL TISSUES APPEARED NORMAL. UTERINE CONTENTS: ONE IMPLANTATIONS 18917 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18918 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18919 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18920 DL 22 p 86 ALL TISSUES APPEARED NORMAL. 18922 DL 22 p 85 ALL TISSUES APPEARED NORMAL. 18923 DL 22 p 87 ALL TISSUES APPPARED NORMAL. 18924 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 1892S DG 26 NP 69 ALL TISSUES APPEARED NORMAL. 18926 DL 14 p 81 LEFT INGUINAL AREA: 14ASS (2.5 CM X ALL OTHER TISSUES APPEARED NORMAL. --------------------------------------------------------------------------------------------------------------- P PREGNANT NP - NOT PREGNANT DS DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION a. Refer to the individual clinicalobeervations table (Table 19) for external observations confirmed at necr b. The fetus had a large, black right!hindlimb and a black tail. Evidence of compression on the abdomen and c. First observed at necropsy. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 20 (PAGE 2): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS ---------------------------------------------------------------------------------------------------------------- DOSAGE GROUP RAT DAY OF PREGNANCY DOSAGES DOSAGE (MG/KG/DAY) NUMBER NECROPSY STATUS ADMINISTERED OBSERVATIONS a --------------------------------------------------------------------------------------------------------------- I 0 (VEHICLE) 18927 DL 14 p 81 18928 DS 72 NP 71 18929 DL 22 p 88 ALL TISSUES ALL TISSUES ALL TISSUES APPEARED APPEARED APPEARED NORMAL. NORMAL. NORMAL. 18930 DG 25 NP 72 ALL TISSUES APPEARED NORMAL. 18931 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18932 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18933 DL 9 p 72 NECROPSY OBSERVATIONS WERE NOT RECOR 18934 DL 22 p 86 ALL TISSUES APPEARED NORMAL. 18935 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18936 DL 22 p 86 ALL TISSUES APPEARED NORMAL. 18937 DL 22 p 88 ALL TISSUES APPEARED NORMAL. 18938 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18939 DL 14 p 77 ALL TISSUES APPEARED NORMAL. 18940 DL 14 p 82 ALL TISSUES APPEARED NORMAL. 18941 DL 22 p 88 ALL TISSUES APPEARED NORMAL. 18942 DL 22 p 88 ALL TISSUES APPEARED NORMAL. ---------------------------------------------------------------------------------------------------------------- P PREGNANT NP - NOT PREGNANT DS DAY OF STUDY DG - DAY OF PRESUMED GESTATION DL = DAY OF LACTATION a. Refer to the individual clinical observations table (Table 19) for external observations confirmed at necrc PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 20 (PAGE 3): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Po GENERATION FEMALE RATS -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP RAT DAY OF PREGNANCY DOSAGES DOSAGE (MG/KG/DAY) NUMBER NECROPSY STATUS ADMINISTERED OBSERVATIONS a -------------------------------------------------------------------------------------------------------------- ii 1.6 18943 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18944 DG 25 NP 71 ALL TISSUES APPEARED NORMAL. 16945 DS 72 NP 71 ALL TISSUES APPEARED NORMAL. 18946 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18947 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18948 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18949 DG 25 NP 71 ALL TISSUES APPEARED NORMAL. 18950 DL 22 p 88 ALL TISSUES APPEARED NORMAL. 18951 DL 22 p 88 ALL TISSUES APPEARED NORMAL. 18952 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18953 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18954 DG 25 NP 69 ALL TISSUES APPEARED NORMAL. 18955 DG 25 NP 69 ALL TISSUES APPEARED NORMAL. 18956 DL 14 p 82 ALL TISSUES APPEARED NORMAL. 18957 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 189se DL 22 p 88 ALL TISSUES APPEARED NORMAL. 18959 DL 22 p 85 ALL TISSUES APPEARED NORMAL. 18960 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18961 DL 22 p 86 ALL TISSUES APPEARED NORMAL. 18962 DG 25 NP 69 ALL TISSUES APPEARED NORMAL. 18963 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18965 DL 22 p 88 ALL TISSUES APPEARED NORMAL. 18966 DS 72 NP 71 ALL TISSUES APPEARED NORMAL. 18967 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18968 DL 22 p 86 ALL TISSUES APPEARED NORMAL. 18969 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18970 DL 22 p 86 ALL TISSUES APPEARED NORMAL. 18971 DL 14 p 81 ALL TISSUES APPEARED NORMAL. 18972 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18973 DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18974 DL 4 p 69 ALL TISSUES APPEARED NORMAL. 18975 DG 25 NP 71 ALL TISSUES APPEARED NORMAL. 18976 DL 22 p 86 ALL TISSUES APPEARED NORMAL. 18977 DL 3 p 69 ALL TISSUES APPEARED NORMAL. UTERINE CONTENTS: 2 FETUSES.B 18978 DG 25 NP 69 ALL TISSUES APPEARED NORMAL. --------------------------------------------------------------------------------------------------------------- P PREGNANT NP - NOT PREGNANT DS DAY OF STUDY DG - DAY OF PRESUMED GESTATION DL = DAY OF LACTATION a. Refer to the individual clinical observations table (Table 19) for external observations confirmed at necr b. Appeared normal for their developmental ages and moderate degree of autolysis at time maternal death occur PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PAGE l)- BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Po GENERATION FEMALE RATS ---------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY ---------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 a 9 10 11 12 13 14 ---------------------------------------------------------------------------------------------------------------- 18901 221. 222. 229. 229. 232. 238. 250. 241. 239. 246. 252. 256. 255. 258 18902 238. 244. 244. 233. 239. 239. 249. 237. 246. 250. 250. 248. 252. -253 18903 227. 233. 232. 240. 237. 247. 257. 253. 246. 252. 260. 262. 256. 265 18904 18905 228. 226. 228. 227. 232. 234. 235. 238. 239. 243. 241. 244. 248. 248 214. 2oa. 218. 224. 223. 221. 236. 229. 226. 223. 230. 233. 236. 231 18906 254. 249. 255. 260. 268. 256. 275. 271. 273. 269. 275. 280. 283. 280 18907 226. 230. 236. 240. 238. 243. 250. 249. 247. 250. 255. 256. 253. 258 18908 233. 222. 226. 221. 234. 233. 238. 234. 237. 232. 236. 240. 236. 237 18909 226. 228, 222. 231. 237. 238. 234. 245. 244. 247. 240. 248. 250. 251 18910 237. 241. 241. 235. 237. 236. 247. 238. 234. 245. 241. 242. 242. 237 18911 234. 223. 237. 238. 239. 234. 249. 249. 249. 244. 253. 262. 251. 247 16912 232. 224. 232. 240. 242. 235. 248. 250. 249. 243. 253. 257. 255. 251 18913 214. 220. 225. 221. 226. 232. 237. 235. 242. 232. 240. 247. 252. 248 18914 240. 241. 250. 255. 255. 253. 266. 272. 270. 267. 273. 272. 278. 277 18915 230. 228. 234. 238. 241. 244. 248. 250. 258. 253. 256. 260. 264. 272 18916 222. 231. 236. 234. 235. 247. 255. 248. 246. 254. 256. 254. 249. 259 18917 219. 225. 230. 233. 231. 241. 247. 243. 244. 250. 257. 256. 258. 257 18918 233. 247. 238. 232. 24S. 253. 256. 239. 248. 251. 253. 240. 255. 260 18919 228. 227. 232. 234. 234. 234. 246. 243. 244. 239. 247. 247. 248. 250 18920 229. 238. 240. 242. 240. 251. 259. 252. 252. 258. 260. 262. 260. 26S 18921 228. 221. 231. 236. 237. 230. 240. 243. 238. 232. 242. 242. 239. 240 18922 235. 243. 246. 239. 243. 243. 252. 251. 252. 254. 260. 261. 258. 262 18923 224. 230. 229. 234. 232. 235. 246. 242. 238. 236. 242. 243. 245. 246 18924 242. 238, 242. 248. 250. 2SO. 259. 256. 260. 255. 267. 267. 268. 260 18925 233. 223. 236. 239. 239. 237. 243. 250. 243. 241. 244. 246. 248. 246 18926 241. 241. 244. 251. 256. 2S4. 257. 256. 257. 260. 261. 265. 268. 268 ---------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PACE 2): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 11 12 13 1 --------------------------------------------------------------------------------------------------------------- 18927 231. 224. 230. 235. 230. 225. 240. 236. 237. 229. 238. 239. 239. 23 18928 245. 234. 242. 248. 251. 246. 260. 256. 263. 249. 252. 260. 256. _25 18929 223. 218. 222. 224. 225. 225. 237. 234. 231. 233. 239. 241. 240. 23 18930 234. 235. 236. 244. 246. 250. 249. 253. 258. 262. 256. 262. 267. 27 16931 234. 228. 240. 246. 247. 244. 264. 258. 257. 251. 260. 261. 262. 25 18932 229. 234. 242. 240. 238. 247. 258. 243. 243. 249. 255. 259. 252. 25 18933 221. 220. 216. 218. 219. 211. 225. 226. 227. 231. 229. 233. 233. 23 18934 234. 241. 240. 232. 246. 250. 256. 247. 253. 260. 265. 258. 267. 27 18935 228. 226. 228. 229. 232. 236. 246. 244. 244. 239. 241. 243. 245. 25 18936 234. 239. 234. 241. 244. 245. 244. 243. 254. 252. 250. 256. 260. 26 18937 224. 226. 225. 230. 232. 234. 236. 235. 238. 239. 241. 240. 240. 24 18938 246. 234. 249. 253. 254. 248. 260. 259. 261. 253. 262. 273. 271. 26 18939 230. 231. 225. 235. 243. 241. 240. 245. 244. 250. 248. 252. 257. 25 18940 243. 238. 238. 247. 255. 259. 256. 258. 265. 265. 259. 270. 278. 27 18941 228. 231. 237. 238. 237. 232. 246. 245. 247. 249. 243. 248. 253. 25 18942 245. 250. 256. 261. 258. 249. 256. 257. 254. 247. 255. 257. 258. 25 --------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PAGE 3): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAY 17 18 19 20 21 22 23 24 25 26 27 28 29 3 --------------------------------------------------------------------------------------------------------------- 18901 257. 259. 265. 260. 260. 266. 267. 267. 265. 270. 274. 277. 275. 28 18902 258. 261. 260. 252. 257. 266. 269. 261. 265. 270. 271. 268. 274. _28 18903 262. 268. 271. 267. 266. 277. 278. 275. 275. 282. 288. 285. 278. 28 18904 254. 255. 250. 257. 255. 258. 2S3. 253. 257. 254. 261. 261. 262. 26 18905 241. 235. 242. 239. 243. 238. 246. 246. 242. 238. 246. 252. 248. 24 18906 293. 289. 300. 300. 301. 299. 307. 306. 308. 298. 309. 316. 305. 30 18907 259. 264. 264. 267. 266. 271. 271. 272. 271. 275. 278. 281. 279. 28 18908 240. 243. 250. 249. 248. 247. 248. 252. 250. 253. 257. 264. 261. 25 18909 257. 260. 256. 251. 260. 262. 268. 255. 268, 267. 267. 266. 264. 26 18910 249. 246. 246. 247. 250. 255. 252. 250. 256. 251. 2Sl. 253. 258. 26 18911 259. 253. 263. 266. 261. 262. 271. 272. 269. 271. 281. 273. 279. 27 18912 262. 260. 265. 272. 271. 269. 266. 273. 278. 281. 279. 275. 284. 28 18913 259. 250. 259. 263. 258. 259. 259. 260. 263. 256. 266. 271. 266. 26 18914 286. 286. 290. 294. 295. 293. 292. 296. 299. 296. 302. 302. 302. 30 18915 278. 277. 270. 279. 280. 285. 278. 281. 289. 290. 284. 290. 293. 29 18916 259. 262. 273. 268. 266. 269. 271. 268. 260. 266. 269. 278. 269. 27 18917 257. 261. 266. 2S6. 262. 269. 268. 266. 265. 268. 274. 275. 276. 27 18918 249. 257. 262. 262. 248. 245. 256. 255. 258. 250. 262. 262. 265. 26 18919 257. 255. 258. 259. 259. 260. 263. 265. 268. 264. 266. 271. 268. 26 18920 262. 270. 269. 265. 267. 270. 275. 271. 271. 275. 277. 271. 270. 27 18921 250. 246. 250. 2S4. 254. 248. 252. 2S4. 254. 248. 260. 262. 262. 25 18922 263. 272. 276. 274. 270. 274. 278. 276. 278. 280. 282. 286. 288. 28 18923 250. 249. 249. 256. 253. 257. 253. 254. 256. 256. 259. 260. 259. 26 18924 276. 276. 284. 286. 286. 282. 289. 291. 287. 283. 296. 298. 293. 29 18925 257. 256. 248. 254. 257. 260. 249. 252. 255. 264. 266. 261. 258. 26 18926 273. 274. 271. 278. 280. 282. 276. 278. 288. 280. 284. 288. 291. 28 --------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PACE 4): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS ---------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY ---------------------------------------------------------------------------------------------------------------- DAY 17 18 19 20 21 22 23 24 25 26 27 28 29 30 ---------------------------------------------------------------------------------------------------------------- 18927 241. 241. 244. 243. 243. 242. 246. 243. 243. 240, 242. 246. 244. 243 18929 265. 262. 268. 271. 269. 266. 276. 273. 277. 268. 280. 284. 278. 277 18929 245. 246. 252. 252. 255. 253. 261. 258. 260. 256. 260. 264. 260. 257 10930 275. 272. 269. 270. 277. 277. 275. 281. 283. 283. 280. 286. 21. 288 18931 264. 261. 270. 265. 273. 268. 266. 268. 270. 270. 278. 280. 278. 278 18932 262. 266. 267. 267. 263. 266. 270. 270. 265. 270. 278. 278. 274. 279 18933 246. 247. 240. 248. 249. 254, 252. 254. 258. 254. 247. 256. 262. 260 18934 280. 284. 281. 276. 288. 292. 290. 283. 294. 296. 295. 292. 297. 304 18935 254. 256. 263. 269. 267. 263. 267. 264. 271. 262. 270. 275. 274. 272 18936 262. 267. 260. 265. 272. 272. 268. 275. 276. 274. 271. 284. 282. 285 18937 240. 246. 250. 250. 247. 246. 249. 249. 254. 247. 246. 249. 252. 256 18938 279. 272. 277. 278. 276. 278. 278. 283. 283. 276. 284. 290. 290. 279 18939 265. 263. 263. 268. 270. 269, 268. 271. 279. 277. 272. 278. 280. 278 18940 286. 286. 279. 282. 293. 293. 279. 285. 301. 294. 284. 295. 300. 302 18941 257. 263. 260. 254. 255. 262. 266. 269. 270. 262. 272. 27S. 275. 274 18942 263. 262. 266. 268. 271. 262. 269. 272. 271. 262. 275. 274. 272. 266 ---------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER! T-6295.13) TABLE 21 (PAGE 5): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAY 33 34 35 36 37 38 39 40 41 42 43a --------------------------------------------------------------------------------------------------------------- 18901 284. 284. 293. 294. 291. 294. 295. 293. 288. 290. b 18902 279. 283. 285. 279. 285. 286. 282. 278. 277. 282. b 18903 282. 292. 296. 295. 291. 297. 296. 294. 287. 295. 299. 18904 267. 269. 265. 268. 271. 272. 270. 271. 271. 272. b 18905 252. 252. 260. 258. 260. 251. 258. 259. 255. 250. b 18906 320. 317. 325. 328. 331. 320. 324. 332. 326. 319. b 18907 287. 287. 299. 288. 286. 287. 292. 291. 282. 284, b 18908 261. 261. 266. 266. 262. 257. 264. 264. 262. 258. b 18909 274. 275. 274. 281. 287, 283. 276. 281. 280. 280. b 18910 260. 264. 273. 273. 269. 263. 269. 269. 268. 267. b 18911 283. 277. 292. 286. 291. 277. 287. 291. 280. 280. b 18912 278. 286. 290. 287. 290. 286. 293. 296. 294. 287. 288. 18913 277. 270. 286. 289. 285. 271. 280. 286. 284. 277. 283. 18914 310. 307. 316. 318. 317. 311. 314. 318. 316. 314. b 18915 297. 299. 296. 301. 304. 302. 299. 305. 310. 308. b 18916 278. 277. 288. 288. 286. 290. 292. 290. 281. 290. 287. 18917 275. 282. 286. 289. 283. 280. 287. 284. 280. 286. b 18918 275. 273. 268. 262. 276. 278. 276. 266. 260, 271. 268. 18919 273. 274. 275. 280. 279. 274. 279. 278. 276. 272. 272. 18920 273. 285. 285. 280. 278. 281. 285. 281. 278. 285. 287. 18921 268. 262. 275. 273. 273. 266. 268. 271. 269. 263. 262. 18922 302. 306. 302. 303. 305. 305. 303. 300. 309. 310. b 18923 265. 267. 273. 268. 269. 266. 273. 273. 268. 271. 268. 18924 297. 298. 303. 307. 305. 297. 307. 311. 307. 301. b 18925 260. 273. 279. 272. 267. 270. 276. 274. 266. 276. b 18926 292. 293. 293. 299. 300. 299. 300. 299. 302. 301. 303. --------------------------------------------------------------------------------------------------------------- DAY - DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. Rat assigned to cohabitation on day 42 of study. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PAGE 6); BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAY 33 34 35 36 37 38 39 40 41 42 43a --------------------------------------------------------------------------------------------------------------- 18927 250. 247. 254. 258. 259. 252. 258. 259. 255. 251. b 18928 283. 281. 292. 297. 289. 285. 292. 294. 289. 284. b 18929 266. 266. 268. 273. 274. 268. 274. 273. 270. 267. b 18930 294. 292. 291. 295. 299. 298. 298. 299. 304. 303. 290. 18931 283. 285. 290. 294. 290. 283. 287. 297. 288. 287. 289. 18932 277. 281. 291. 291. 286. 288. 290. 290. 286. 292. b 18933 268. 264. 261. 262. 272. 272. 268. 271. 278. 269. b 18934 303. 310. 312. 305. 315. 315. 315. 308. 318. 323. b 18935 278. 272. 285. 286. 285. 283. 286. 285. 285. 278. 278. 18936 286. 289. 285. 291. 293. 262. 263. 263. 263. 267. 271. 18937 255. 260. 259. 254. 264. 267. 263. 262. 259. 260. b 18938 297. 290. 293. 304. 303. 298. 296. 296. 300. 298. 294. 18939 287. 285. 280. 294. 290. 290. 289. 291. 299. 292. b 18940 308. 303. 298. 302. 314. 307. 298. 305. 310. 313. 299. 18941 276. 283. 284. 278. 284. 286. 289. 285. 285. 284. 288. 18942 275. 273. 282. 282. 287. 276. 282. 282. 281. 278. b -------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. Rat assigned to cohabitation on day 42 of study. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR-S STUDY NUMBER: T-6295.13) TABLE 21 (PAGE 7)! BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 11 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 11 12 13 1 --------------------------------------------------------------------------------------------------------------- 18943 218. 217. 227. 226. 232. 224. 234. 236. 234. 229. 238. 240. 236. 23 18944 256. 266. 271. 270. 263. 267. 266. 263. 261. 268. 270. 268. 262. 27 18945 234. 231. 229. 238. 238. 244. 236. 242. 246. 247. 240. 249. 253. 24 18946 236. 238. 245. 248. 238. 236. 241. 244. 242. 238. 246. 247. 248. 24 18947 211. 221. 228. 230. 228. 239. 241. 245. 239. 246. 250. 253. 252. 25 18948 250. 246. 261. 257. 261. 255. 264. 268. 261. 250. 253. 270. 262. 26 18949 228. 221. 216. 224. 231. 229. 226. 232. 237. 237. 233. 245. 246. 24 18950 234. 230. 236. 243. 245. 239. 249. 251. 257. 248. 257. 262. 264. 25 18951 227. 227. 225. 229. 233. 237. 229. 236. 239. 232. 228. 232. 240. 23 18952 238. 244. 249. 251. 244. 259. 258. 261. 255. 265. 268. 269. 262. 27 18953 235. 228. 237. 242. 241. 237. 242. 244. 240, 240. 249. 250. 248. 24 18954 238. 233. 234. 241. 245. 247. 241. 249. 250. 249. 247. 245. 251. 25 18955 225, 225. 220. 228. 233. 236. 227. 233. 231. 234. 232. 240. 242. 23 18956 241. 239. 235. 243. 242. 246. 239. 247. 251. 250. 252. 255. 261. 26 18957 226. 237. 242. 240. 234. 249. 249. 249. 242. 252. 258. 254. 254. 25 18958 232. 234. 229. 241. 247. 246. 245. 254. 258. 259. 251. 260. 264. 26 18959 222. 219. 226. 228. 231. 230. 230. 241. 240. 244. 240. 237. 247. 24 18960 246. 245. 255. 254. 262. 261. 269. 272. 272. 272. 275. 279. 277. 27 18961 222. 225. 226. 222. 234, 236. 235. 231. 236. 241. 244. 237. 245. 24 18962 225. 231. 232. 241. 236. 246. 252. 253. 252. 259. 260. 259. 258. 26 18963 217. 227. 230. 226. 221. 233. 241. 238. 233. 240. 242. 241. 232. 24 18964 237. 240. 240. 234. 244. 247. 244. 237. 243. 240. 242. 242. 248. 25 1896S 224. 224. 224. 232. 230. 233. 229. 236. 232. 232. 236. 237. 238. 23 18966 238. 239. 244. 242. 2S3. 246. 248. 254. 254. 259. 252. 252. 256. 26 18967 226. 223. 235. 238. 236. 233. 246. 251. 249. 244. 2S7. 259. 258. 25 18968 239. 246. 247. 243. 255. 2S6. 259. 254. 266. 264. 268. 262. 270. 27 --------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PAGE 8): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 11 1.6 MG/KG/DAY --------------------- ----------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 a 9 10 11 12 13 1 --------------------------------------------------------------------------------------------------------------- 18969 229. 225. 235. 242. 237. 236. 243. 246. 247. 244. 252. 259. 256. 25 18970 224. 234, 235. 238. 233. 240. 248. 245. 243. 245. 254. 254. 249. 25 18971 224. 222. 231. 233, 23S. 234. 243. 243. 245. 239. 245. 252. 250. 24 18972 234. 239. 245. 247. 243. 250. 253. 250. 254. 259. 260. 264. 262. 26 18973 230. 227. 236. 240. 238. 236. 243. 247. 244. 242. 249. 252. 249. 24 18974 236. 240. 250. 250. 246. 254. 262. 263. 2S9. 263. 271. 272. 269. 27 18975 234. 231. 245. 242. 246. 242. 254. 256. 257. 250. 263. 266. 266. 26 18976 232. 240. 247. 249. 246. 253. 246. 229. 242. 241. 241. 243. 249. 25 18977 222. 229. 226. 224. 234. 236. 236. 235. 238. 242. 243. 240. 245. 24 18978 234. 244. 243. 240. 252. 252. 254. 251. 255. 253. 254. 252. 261. 26 --------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PAGE 9): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS ---------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 11 1.6 MG/KG/DAY ---------------------------------------------------------------------------------------------------------------- DAY 17 18 19 20 21 22 23 24 25 26 27 28 29 30 ---------------------------------------------------------------------------------------------------------------- 18943 240. 23S. 241. 244. 243. 246. 249. 252. 250. 248. 252. 252. 250. 249 18944 272. 272. 277. 274. 276. 280. 287. 284. 278. 287. 290. 290. 290. 288 18945 253. 247. 247. 251. 255. 251. 246. 250. 257. 253. 250. 253. 254. 256 18946 253. 256. 260. 263. 260. 264. 261. 269. 266. 264. 269. 272. 272. 269 18947 258. 269. 268. 265. 262. 269. 268. 265. 262. 264. 265. 268. 262. 269 18948 266, 260. 266. 267. 268. 271. 273. 278. 279. 272. 280. 279. 279. 274 18949 250. 247. 247. 250. 259. 252, 253. 257. 259. 258. 253. 262. 264. 258 18950 266. 261. 270. 270. 270. 266. 268. 268. 271. 266. 270. 275. 272. 267 18951 243. 238. 236. 242. 243. 244. 241. 246. 249. 246. 246. 251. 254. 256 18952 270. 283. 287. 277. 268. 286. 284. 283. 276. 283. 287. 289. 282. 292 18953 257. 254. 254. 258. 262. 259. 264. 267. 269. 269. 278. 278. 279. 274 18954 253. 256. 261. 259. 258. 257. 258. 261. 258. 257. 266. 261. 261. 259 18955 243. 244. 240. 246. 252. 253. 251. 255. 258. 258. 254. 263. 263. 263 18956 268. 266. 263. 269. 267. 267. 265. 268. 272. 270. 271. 272. 280. 276 18957 254. 264. 264. 260. 255. 266. 263. 262. 258. 265. 268. 265. 263. 273 18958 274. 270. 265. 269. 275. 274. 268. 272. 275. 276. 268. 278. 280. 279 18959 243. 249. 253. 253. 250. 250. 252. 254. 2S6. 250. 250. 255. 257. 256 18960 285. 282. 296. 288. 290. 288. 293. 294. 296. 291. 299. 303. 296. 297 18961 251. 253. 252. 249. 252. 258. 2S2. 244. 252. 257. 258. 258. 261. 262 18962 269. 273. 276. 272. 272. 27B. 271. 273. 269. 282. 287. 287. 283. 290 18963 241. 252. 253. 249. 241. 254. 256. 254. 248. 251. 259. 257. 248. 259 18964 255. 258. 258. 250. 261. 260. 258. 246. 257. 263. 263. 261. 263. 262 18965 242. 239. 237, 246. 242. 242. 238. 244. 242. 244. 242. 248. 245. 247 18966 259. 265. 267. 268. 270. 267. 270. 272. 269. 271. 274. 275. 274. 276 18967 262. 261. 264. 266. 266. 266. 274. 275. 273. 272. 274. 276. 277. 276 18968 279. 281. 280. 270. 275. 280. 276. 273. 285. 279. 284. 278. 286, 289 ---------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014! ORAL (GAVAGE) CROSS-FOSTERINO STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PAGE 10.): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE PATS ---------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 11 1.6 MG/KG/DAY ---------------------------------------------------------------------------------------------------------------- DAY 17 18 19 20 21 22 23 24 25 26 27 28 29 30 ---------------------------------------------------------------------------------------------------------------- 18969 260. 258. 267. 264. 265. 266. 271. 274. 271. 271. 274. 276. 279. 270 18970 256. 261. 265. 264. 258. 268. 271. 266. 263. 272. 274. 272. 269. 275 18971 253. 2SO. 25S. 259. 262. 262. 260. 261. 264. 263. 268. 268. 267. 264 18972 271. 273. 278. 276. 276. 279. 284. 282. 201. 278. 285. 289. 286. 288 18973 260. 254. 257. 259. 259. 261. 262. 262. 265. 262. 266. 270. 267. 261 18974 266. 274. 274. 276. 272. 280. 284. 282. 279. 281. 289. 283. 278. 288 18975 281. 273. 280. 286. 285. 282. 202. 291. 290. 287. 290. 296. 291. 288 18976 258. 260. 262. 261. 260. 271. 268. 264. 265. 271. 270. 270. 267. 280 18977 253. 254. 250. 248. 2S8. 263. 255. 252. 261. 263. 262. 255. 265, 264 18978 266. 268. 264. 261. 274. 275. 266. 266. 273. 273. 270. 269. 272. 27 --------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 410-014: OPAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PAGE 11): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP Il 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAY 33 34 35 36 37 38 39 40 41 42 43a --------------------------------------------------------------------------------------------------------------- 18943 257. 252. 263. 262. 259. 250. 258. 257. 251. 248. 2S2. 18944 296. 295. 295. 304. 309. 302. 300. 305. 303. 298. 292. 18945 257. 255. 257. 259. 267. 264. 256. 261. 262. 261. 253. 18946 271. 278. 275. 280. 281. 274. 277. 283. 282. 280. 281. 18947 265. 274. 276. 274. 267. 275. 276. 273. 267. 273. 271. 18948 282. 284. 288. 289. 286. 280. 280. 290. 281. 279. 288. 18949 271. 263. 260. 269. 272. 265. 267. 270. 271. 268. 263. 18950 270. 266. 265. 273. 276. 272. 266. 270. 271. 271. 269. 18951 255. 256. 251. 260. 258. 255. 250. 253. 252. 252. 245. 18952 288. 295. 301. 299. 291. 295. 299. 294. 284. 291. 288. 18953 18954 283. 268. 280. 270. 283. 268. 287. 264. 288. 266. 280. 265. 280. 266. 286. 262. 287. 258. 283. 261. 280. 254. 18955 262. 263. 261. 265. 266. 258. 257. 259. 259. 259. 253. 18956 286. 281. 283. 283. 287. 283. 279. 282. 288. 283. 275. 18957 271. 275. 276. 273. 271. 275. 277. 280. 272. 278. 275. 18958 285. 280. 279. 282. 284. 284. 279. 285. 284. 281. 270. 18959 261. 261. 264. 264. 266. 272. 268. 260. 270. 274. 271. 18960 309. 303. 314. 315. 310. 307. 313. 313. 308. 307. 310. 18961 264. 270. 270. 265. 270. 269. 264. 262. 266. 270. 269. 18962 288. 288. 293. 288. 290. 289. 293. 289. 287. 291. 290. 18963 253. 263. 267. 266. 263. 265. 265. 263. 256. 258. 259. 18964 267. 271. 268. 265. 267. 266. 262. 258. 260. 261. 252. 18965 252. 246. 247. 250. 248. 249. 246. 246. 252. 248. 240. 18966 280. 280. 278. 274. 284. 282. 282. 279. 282. 287. 281. 18967 283. 284. 288. 292. 294. 287. 292. 291. 290. 285. 288. 18968 292. 299. 296. 286. 295. 292. 294. 289. 293. 299. 286. --------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN PATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PAGE 12); BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP Il 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAY 33 34 35 36 37 38 39 40 41 42 43a --------------------------------------------------------------------------------------------------------------- 18969 276. 275. 286. 286. 286. 281. 288. 291. 287. 285. 284. 18970 272. 279. 285. 282. 277. 280. 281. 281. 274. 281. 282. 18971 272. 270. 280. 280. 277. 271. 276. 276. 274. 275. 275. 18972 291. 289. 295. 297. 296. 293. 297. 298. 295. 289. 291. 18973 268. 268. 276. 276. 277. 269. 274. 275. 270. 269, 275. 18974 282. 284, 285. 282. 284. 286. 292. 284. 278. 286. 288. 18975 302. 295. 308. 309. 305. 299. 301. 302. 305. 301. 301. 18976 275. 281. 279. 281. 278. 279. 283. 278. 274. 280. 280. 18977 271. 272. 276. 267. 274. 276. 275. 267. 272. 276. 273. 18978 278. 277. 275. 272. 284. 276. 276. 265. 276. 276. 275. --------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 22 (PAGE 1): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS ----------------------- ---------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY ---------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 10 ---------------------------------------------------------------------------------------------------------------- 18901 p 18902 NP 18903 P 18904 P 18905 NP 293. 288. 296. 276. MATING 309. 311. 300. 299. 307. 313. 280. 286. NOT CONFIRMED 312. 303. 316. 292. 321. 308. 325. 294. 316. 314. 329. 299. 323. 316. 333. 301. 327. 318. 330. 307. 326. 317. 335. 304. 330. 321. 342. 308. 328. 320. 346. 314. 18906 p 334. 348. 359. 366. 370. 373. 378. 377. 384. 386. 392. 18907 P 301. 317. 320. 319. 318. 330. 329. 327. 333. 336. 342. 18908 P 276. 287. 262. 287. 292. 291. 304. 302. 307. 308. 316. 18909 p 290. 300. 305. 304. 308. 310. 317. 320. 321. 325. 330. 18910 p 270. 280. 287. 286. 293. 295. 299. 301. 304. 306. 311. 18911 P 293. 308. 308. 310. 318. 316. 322. 327. 330. 335. 333. 18912 P 302. 312. 316. 319. 321. 328, 328. 337. 335. 340. 348. 18913 P 291. 299. 306. 306. 300. 300. 302. 314. 309. 313. 326. 18914 NP 307. 317. 332. 329. 335. 342. 350. 350. 350. 356. 36S. 18915 P 320. 326. 327. 335. 335. 340. 346. 345. 354. 358. 367. 18916 P a 286. 296. 296. 296. 310. 306. 308. 310. 317. 323. 321. 18917 P 298. 308. 305. 308. 314. 317. 323. 326. 328, 330. 334. 18918 P 279. 292. 293. 299. 301. 312. 319. 322. 321. 331. 332. 18919 P 286. 300. 300. 304. 306. 315. 320. 320. 329. 326. 341. 18920 P 294. 299. 303. 307. 310. 31S. 316. 319. 317. 322. 327. 18921 NP 269. 266. 262. 265. 267. 263. 259. 262. 263. 263. 260. 10922 P 310. 320. 318. 325. 329. 336. 341. 345. 352. 357. 367. 18923 P 276. 282. 287. 291. 296. 298. 299. 302. 301. 305. 310. 18924 P 323. 333. 341. 346. 355. 347. 351. 360. 356. 360. 364. 18925 NP 284. 278. 284. 268. 284. 275. 289. 285. 287. 280. 283. ---------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Dam 18916 did not deliver a litter; only one dead fetus was present in utero on day 25 of gestation. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NLTMBER: T-6295.13) TABLE 22 (PAGE 2); MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE PATS ----------- -------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 10 --------------------------------------------------------------------------------------------------------------- 18926 P 18927 P 18928 NP 317. 273. MATING 320. 327. 288. 293. NOT CONFIRMED 328. 298. 332. 303. 332. 304. 337. 304. 340. 315. 344. 313. 345. 355 317. j20 18929 P 292. 302. 305. 308. 320. 317. 323. 330. 329. 33S. 349 18930 NP 312. 324. 329. 332. 334. 334. 336. 338. 337. 344. 343 18931 P 298. 300. 310. 314. 320. 322. 330. 336. 331. 336. 348 18932 P 290. 294. 304. 307. 312. 315. 316. 316. 321. 328. 330 18933 P 288. 289. 293. 300. 306. 303. 304. 312. 319. 322. 330 18934 P 315. 327. 336. 340. 341. 351. 349. 354. 361. 368. 368 18935 P 295. 292. 297. 306. 319. 321. 330. 326. 327. 330. 336 18936 P 270. 272. 279. 278. 277, 278. 277. 291. 305. 313. 312 18937 P 265. 274. 284. 289. 294. 300. 296. 301. 304. 300. 309 18938 P 306. 317. 328. 329. 328. 334. 335. 341. 343. 351. 358 18939 P 302. 310. 313. 323. 324. 326. 326. 330, 334. 336. 346 18940 P 329. 333. 340. 346. 357. 369. 363. 360. 362. 370. 376 18941 P 300. 308. 31S. 326. 327. 333. 33S. 338. 343. 347. 356 18942 P 294. 302. 305. 312. 315. 321. 321. 330. 325. 328. 337 --------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT DAY = DAY OF PRESUMED GESTATION (VALUES EXCLUDED FROM AVERAGES) ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: OPAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 22 (PAGE 3): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAY 13 14 is 16 17 18 19 20 21 22 23 --------------------------------------------------------------------------------------------------------------- 18901 p 18902 NP 18903 P 18904 P 18905 NP 344. 353. 358. 374. 379. 400. 417. 436. 455. 325. 315. 310. 309. 309. 307. 300. 304. 313. 314. io6 361. 367. 380. 392. 402. 426. 452. 470. 488. 322. 326. 335. 356. 371. 387. 406. 427. 450. 304. MATING NOT CONFIRMED 18906 P 422. 414. 426. 440. 460. 483. 499. 523. 526. 18907 P 354. 363, 376. 380. 396. 414. 436. 4S6. 478. 18908 p 333. 339. 348. 363. 377. 397. 413. 443. 459. 18909 p 18910 p 349. 320. 356. 319. 362. 324. 378. 333. 394. 342. 395. 346. 414. 355. 427. 357. 445. 369. 429. 18911 p 355. 3S4. 365. 374. 392. 409. 424. 455. 464. 329. 18912 P 366. 373. 374. 388. 406. 427. 444. 467. 480. 18913 P 343. 343. 352. 368. 383. 404. 409. 434. 443. 457. 18914 NP 340. 347. 341. 342. 346. 349. 346. 340. 350. 347. 351 18915 P 389. 397. 406. 426. 436. 452. 478. 491. 525. 18916 P a 331. 339. 340. 334. 334. 340. 347. 341. 341. 336. 322 18917 P 347. 358. 365. 372. 386. 396. 412. 428. 444. 438. 10918 p 18919 p 359. 366. 354. 367. 366. 366. 369. 382. 378. 394. 395. 413. 413. 434. 425. 454. 442. 463. 431. 18920 P 348. 356. 364. 372. 383. 406. 428. 434. 451. 18921 NP 18922 P 265. 374. 259. 383. 268. 394. 267. 404. 263. 415. 258. 431. 263. 442. 270. 271. 268. 277 465. 477. 18923 P 32S. 333. 341. 348. 359. 376. 394. 405. 412. 18924 P 393. 38B. 394. 396. 403. 421. 442. 468. 476. 18925 NP 280. 288. 294. 289. 287. 286. 267. 298. 298. 294. 292 --------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT DAY = DAY OF PRESUMED GESTATION (VALUES EXCLUDED FROM AVERAGES) ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Dam 18916 did not deliver a litter; only one dead fetus was present in utero on day 25 of gestation. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN PATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 22 (PAGE 4): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAY 13 14 15 16 17 18 19 20 21 22 23 --------------------------------------------------------------------------------------------------------------- 18926 P 373. 370. 375. 389. 406. 420. 443. 450. 465. 18927 P 339. 339. 336. 353. 364. 385. 402. 429. 443. 18928 NP 18929 P 18930 NP 18931 P 18932 P 18933 P 18934 P 18935 P 18936 P HATING NOT CONFIRMED 370. 375. 373. 382. 405. 423. 437. 470. 478. 477. 338. 329. 329. 335. 335. 328. 334. 335. 327. 322. 336 357. 357. 360. 371. 389. 405. 405. 428. 434. 348. 354. 366. 368. 383. 398. 421. 434. 457. 449. 337. 340. 350. 370. 384. 396. 421. 442. 468. 384. 385. 401. 410. 413. 434. 455. 475. 480, 358. 359. 370. 382. 392. 417. 425. 450. 457. 319. 330. 343. 352. 357. 370. 389. 398. 413. 18937 P 18938 P 323. 378. 324. 382. 328. 385. 343. 401. 359. 417. 374. 438. 394. 446. 418. 472. 427. 474. 433. 18939 P 358. 360. 366. 376. 389, 39S. 409. 428. 462. 18940 P 390. 392. 393. 408. 425. 435. 456. 468. 469. 18941 P 372. 370. 377. 393. 400. 424. 436. 466. 470. 18942 P 353. 364. 365. 373. 386. 401. 420. 437. 445. 310. - --------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 22 (PAGE 5): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- PAT # DOSAGE GROUP 11 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 10 --------------------------------------------------------------------------------------------------------------- 18943 P 18944 NP 18945 NP 261. 307. MATING 272. 270. 319. 322. NOT CONFIRMED 272. 326. 274. 326. 276. 322. 276. 334. 276. 340. 282. 329. 282. 295 333. i40 18946 P 18947 P 18948 P 18949 NP 18950 p 18951 p 18952 P 18953 P 18954 NP 18955 NP 18956 P 18957 P 18958 P 18959 P 18960 P 18961 p 18962 NP 18963 P 18964 P 18965 P 18966 NP 291. 295. 302. 303. 304. 298. 311. 323. 314. 315. 320 282. 286. 293. 292. 297. 300. 304. 309. 313. 318. 321 292. 300. 304. 302. 313. 314. 317. 327. 325. 329. 342 278. 279. 280. 284. 284. 280. 279. 287. 283. 277. 278 287. 290. 294. 296. 299. 296. 303. 300. 309. 307. 317 264. 264. 272. 273. 273. 275. 280. 284. 285. 289. 294 296. 307. 307. 310. 316. 319. 323. 320. 330. 329. 331 295. 305. 306. 309. 304. 308. 316. 316. 320. 320. 329 266. 268. 266. 261. 258. 263. 265. 263. 261. 265. 263 261. 266. 267. 264. 269. 272. 268. 264. 267. 271. 271 288. 296. 298. 293. 295. 290. 296. 299. 303. 305. 309 289. 288. 293. 295. 296. 297. 301. 301. 308. 313. 317 289. 304. 306. 308. 311. 310. 310. 313. 316. 322. 330 270. 273. 280. 283. 280. 287. 284. 290. 293. 296. 301 316. 331. 327. 336. 338. 334. 337. 341. 338. 346. 352 273. 279. 201. 291. 298. 294. 302. 304. 302. 304. 314 290. 283. 287. 288. 282. 281. 282. 284. 280. 275. 281 266. 268. 277. 281. 288. 292. 293. 296. 302. 305. 313 259. 267. 272. 272. 272. 276. 279. 284. 291. 287. 294 264. 268. 269. 271. 272. 278. 275. 280. 281. 285. 293 MATING NOT CONFIRMED 18967 P 285. 282. 299. 303. 307. 302. 301. 306. 309. 31Q. 323 --------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NLTMBER: T-6295.13) TABLE 22 (PAGE 6): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - FO GENERATION FEMALE RATS -------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 11 1.6 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 1 -------------------------------------------------------------------------------------------------------------- 18968 p 18969 P 18970 P 18971 P 18972 P 18973 P 18974 P 18975 NP 302. 302. 307. 306. 311. 310. 318. 328. 327. 331. 32 294. 306. 306. 305. 310. 311. 310. 322. 316. 322. 33 286. 292. 295. 297. 294. 298. 301. 300. 310. 309. 31 282. 288. 288. 290. 291. 303. 300. 309. 303. 304. 31 298. 309. 309. 314. 315. 315. 321. 324. 335. 328. 33 268. 279. 276. 282. 280. 288. 288. 293. 291. 294. 30 297. 305. 305. 306. 312. 319. 319. 319. 330. 328. 33 313. 315. 318. 324. 315. 316. 325. 330. 326. 326. 33 18976 P 292. 296. 295. 294. 302. 306. 306. 311. 318. 320. 32 18977 P 18978 NP 280. 285. 283. 289. 294. 299. 308. 315. 316. 315. 32 283. 285. 284. 288. 290. 292. 296. 298. 302. 301. 29 -------------------------------------------------------------------------------------------------------------- P = PREGNANT NP - NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 22 (PAGE 7): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS -------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP ir 1.6 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAY 13 14 15 16 17 18 19 20 21 22 2 -------------------------------------------------------------------------------------------------------------- 18943 P 18944 NP 18945 NP 302. 307. 324. 339. 354. 374. 394. 417. 424. 353. 336. 334. 327. 328. 326. 321. 325. 327. 325. 32 MATING NOT CONFIRMED 18946 P 18947 P 18948 P 18949 NP 18950 P 18951 P 18952 P 18953 P 18954 NP 18955 NP 18956 P 18957 P 18958 P 18959 P 18960 P 18961 P 18962 NP 18963 P 18964 P 18965 P 18966 NP 335. 334. 343. 365. 375. 394. 418. 444. 450. 327. 342. 346. 356. 374. 394. 405. 412. 431. 362. 359. 359. 385. 402. 424. 443. 458. 470. 279. 281. 275. 280. 272. 281. 277. 280. 271. 272. 27 318. 335. 341. 355. 375. 394. 410. 419. 442. 306. 309. 317. 330. 346. 359. 375. 393. 406. 340. 354. 359. 364. 386. 402. 420. 430. 446. 348. 351. 349. 366. 370. 401. 413. 444. 442. 255. 265. 264. 265. 259. 255. 263. 266. 254. 266. 25 273. 272. 270. 272. 268. 266. 296. 269. 273. 274. 26 320. 324. 330. 337. 353. 366. 380. 395. 412. 416. 330. 338. 34S. 350. 369. 389. 401. 413. 420. 346. 349. 356. 367. 38S. 403. 421. 426. 443. 320. 320. 332. 341. 351. 361. 376. 394. 406. 367. 362. 371. 388. 406. 425. 438. 449. 461. 333. 328. 340. 352. 357. 382. 401. 417. 436. 279. 280. 284. 284. 285. 288. 288. 290. 288. 293. 29 329. 339. 345. 355. 365. 380. 378. 391. 406. 304. 309. 319. 323. 338. 361. 374. 391. 399. 310. 316. 332. 345. 371. 384. 407. 411. MATING NOT CONFIRMED 18967 P 345. 349. 352. 368. 388. 406. 425. 445. 457. -------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 416-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 22 (PAGE 8): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS ---------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 11 1.6 MG/KG/DAY ---------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAY 13 14 is 16 17 18 19 20 21 22 23 ---------------------------------------------------------------------------------------------------------------- 18968 P 18969 P 342. 34S. 351. 350. 366. 364. 373. 372. 387. 386. 407. 415. 426. 434. 447. 459. 452. 472. 18970 P 327. 333. 334. 346. 362. 374. 391. 406. 406. 18971 P 18972 P 322. 365. 320. 365. 324. 365. 319. 383. 332. 396. 340. 422. 345. 437. 354. 460. 355. 468. 356. 318. 18973 P 321. 321. 329. 341. 355. 387. 398. 422. 420. 18974 P 354. 367. 374. 378. 396. 410. 433. 440. 469. 18975 NP 18976 P 18977 P 326. 338. 341. 315. 343. 345. 316. 352. 358. 324. 360. 370. 317. 372. 380. 313. 389. 394. 318. 404. 414. 329. 423. 430. 309. 456. 461. 309. 297. 18978 NP 278. 281. 291. 284. 274. 287. 291. 288. 283. 295. 294. ---------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT DAY = DAY OF PRESUMED GESTATION (VALUES EXCLUDED FROM AVERAGES) ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: ORAL IGAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 23 (PAGE 1): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP I a 0 (VEHICLE) MG/KG/DAY SUBSET A --------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 11 ------------------------------------------------------------------------------------------------------------- :-- 18903 353. 342. 337. 333. 338. 341. 336. 345. 346. 365. 363 18907 337. 334. 329. 328. 345. 338. 346. 341. 353. 366. 357 18908 318. 311. 324. 321. 318. 319. 328. 336. 339. 342. 341 18909 319. 319. 317. 321. 337. 336. 347. 362. 345. 359. 350 18918b 300. 309. 321. 341. 345. 343. 353. 360. 350. 356. 347 18920 325. 325. 320. 319. 338. 339. 345. 342. 307.c 345. 360 18924 351. 343. 318. 356. 361. 360. 369. 368. 370. 379. 387 18929 330. 337. 344. 348. 353. 354. 350. 354. 365. 371. 356 18934 18936b 18937 362. 353. 358. 365. 365. 373. 363. 369. 367. 335. 369 315. 302. 310. 314. 303. 306. 326. 335. 332. 343. 333 294. 303. 304. 324. 328. 340. 344. 358. 361. 342. 342 18941 3S4. 356. 348. 358. 358. 360. 356. 357. 365. 371. 383 18942 340. 332. 337. 320. 345. 347. 340. 354. 347. 344. 350 --------------------------------------------------------------------------------------------------------------- DAY 14 is 16 17 18 19 20 21 22 --------------------------------------------------------------------------------------------------------------- 18903 374. 386. 378. 388. 383. 399. 390. 377. 364. 18907 355. 346. 353. 370. 358. 351. 355. 349. 358. 18908 364. 370. 368. 368. 367. 379. 374. 379. 372. 18909 353. 373. 371. 384. 373. 376. 379. 386. 353. 18918 377. 363. 374. 382. 386. 388. 397. 365. 360. 18920 360. 371. 366. 363. 367. 369. 374. 370. 362. 18924 370. 388. 404. 394. 403. 396. 398. 389. 365. 18929 383. 388. 383. 390. 394. 380. 379. 380. 354. 18934 370. 380. 381. 390. 381. 381. 372. 361. 375. 18936 330. 350. 348. 352. 313. 335. 326. 349. 353. 18937 381. 371. 377. 368. 377. 369. 392. 384. 346. 18941 382. 383. 375. 376. 403. 391. 374. 371. 373. 18942 366. 378. 383. 372. 361. 371. 372. 364. 360. --------------------------------------------------------------------------------------------------------------- DAY - DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. Litter was not croBB-fostered; values excluded from group averages. c. Water access was interrupted; value excluded from group averages. PROTOCOI, 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 23 (PAGE 2): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Po GENERATION FEMALE PATS ------------------------------------------------------------------------------------------------------------ RATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP I a 0 (VEHICLE) MG/KG/DAY SUBSET B -------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 1 -------------------------------------------------------------------------------------------------------------- 18901 313. 338. 343. 351. 366. 368. 370. 355. 359. 375. 37 18904 311. 314. 313. 314. 330. 345. 353. 362. 353. 341. 31 18906 399. 394. 407. 420. 426. 420. 417. 391. 405. 430. 41 18912 347. 355. 355. 361. 363. 362, 368. 374. 376. 383. 36 18917 325. 331. 338. 343. 349. 352. 357. 362. 362. 370. 36 18919 338. 324. 344. 355. 3S4. 357. 357. 349. 369. 359. 35 18922 366. 362. 370. 363. 367. 341. 372. 377. 371. 374. 37 18923 280. 292. 300. 307. 314. 317. 330. 323. 330. 344. 34 18931 324. 322. 331. 340. 345. 358. 356. 361. 362. 354. 35 18932 320. 322. 337. 344. 342. 334. 341. 348. 355. 359, 35 18935 320. 313. 320. 316. 339. 334. 319. 327. 328. 323. 32 18938 338. 339. 346. 346. 3Sl. 366. 375. 370. 382. 378. 38 -------------------------------------------------------------------------------------------------------------- DAY 14 is 16 17 18 19 20 21 22 -------------------------------------------------------------------------------------------------------------- 18901 384. 376. 375. 372. 388. 385. 395. 380. 350. 18904 315. 333. 343. 350. 352. 352. 351. 358. 356. 18906 436. 438. 456. 457. 441. 443. 448. 437. 406. 18912 379. 382. 372. 378. 380. 389. 398. 385. 345. 18917 370. 366. 377. 364. 374. 372. 383. 377. 376. 18919 372. 380. 374. 374. 380. 385. 366. 380. 375, 18922 377. 387. 389. 385. 389. 390. 392. 401. 420. 18923 331. 339. 340. 343. 339. 346. 334. 334. 324. 18931 366. 367. 363. 362. 372. 373. 375. 370. 358. 18932 353. 352. 367. 362. 375. 363. 368. 353. 368. 18935 351. 343. 346. 352. 348. 352. 333. 340. 349. 18938 381. 388. 397. 390. 395. 397. 383. 365. 362. -------------------------------------------------------------------------------------------------------------- DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Vehicle control group dams cross-fostered with vehicle control litters. b. Water access was interrupted; value excluded from group averages. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 23 (PAGE 3): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP II a 1.6 MG/KG/DAY SUBSET C --------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 11 --------------------------------------------------------------------------------------------------------------- 18943 18947 291. 297. 307. 310. 325. 320. 324. 322. 326. 331. 316 324. 318. 325. 322. 334. 338. 326. 344. 348. 344. 347 18948 345. 344. 343. 341. 346. 350. 353. 364. 357. 363. 363 18957 318. 316. 313. 312. 317. 326. 325. 323. 332. 345. 337 18960 345. 349. 341. 352. 351. 354. 361. 364. 380. 376. 380 18964 313. 302. 313. 312. 319. 304. 317. 325. 334. 330. 327 18967 331. 327. 335. 342. 336. 331. 338. 361. 345. 360. 348 18968 331. 327. 331. 332. 324. 340. 348. 344. 337. 347. 358 18969 336. 334. 334. 338. 342. 339. 352. 347. 358. 351. 354 18970 324. 311. 316. 324. 326. 329. 332. 345. 345. 359. 348 18973 296. 292. 294. 302. 300. 290. 300. 304. 312. 318. 317 18974 360. 353. 338. 336. NO SURVIVING PUPS ON DAY 4 OF LACTATION --------------------------------------------------------------------------------------------------------------- DAY 14 15 16 17 le 19 20 21 22 --------------------------------------------------------------------------------------------------------------- 18943 344. 334. 362. 358. 353. 356. 289.b 322. 339. 18947 359. 359. 368. 376. 372. 368. 367. 371. 339. 18948 374. 378. 385. 372. 380. 378. 326. 292.b 353. 18957 349. 361. 357. 360. 366. 366. 353. 352. 350. 18960 381. 380. 380. 395. 399. 397. 407. 396. 390. 18964 329. 343. 348. 348. 353. 351. 363. 361. 329. 18967 346. 374. 374. 379. 383. 383. 383. 357. 366. 18968 350. 364. 363. 377. 381. 373. 388. 385. 374. 18969 360. 369. 365. 371. 372. 371. 361. 368. 376. 18970 3SO. 358. 364. 374. 370. 371. 376. 344.b 343. 18973 316. 314. 315. 324. 327. 332. 332. 317. 343. 18974 NO SURVIVING PUPS ON DAY 4 OF LACTATION --------------------------------------------------------------------------------------------------------------- DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day b. Water access was interrupted; value excluded from group averages. dosage group litters. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 23 (PAGE 4): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS ---------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING ---------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP II a 1.6 MG/KG/DAY SUBSET D ---------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 11 ---------------------------------------------------------------------------------------------------------------- 18946b 325. 321. 320. 322. 329. 334. 342. 320. 343. 346. 352. 18950 332. 326. 319. 317. 328. 344. 332. 342. 351. 335. 337. 18951 299. 298. 300. 300. 307. 305. 305. 309. 311. 311. 309. 18952 328. 331. 333. 335. 350. 355. 350. 358. 355. 360. 360. 18953 325. 320. 323. 318. 316. 315. 321. 332. 327. 339. 348. 18958 333. 327. 327. 324. 316. 316. 321. 326. 329. 331. 329. 18959 307. 295. 291. 290. 291. 306. 315. 323. 318. 309. 318. 18961 327. 311. 315. 286. 306. 320. 320. 327. 324. 330. 341. 18963 298. 293. 298. 296. 309. 308. 311. 324. 324. 328. 340. 18965 286. 295. 305. 295. 308. 321. 323. 328. 330. 332. 322. 18972 349. 344. 3Sl. 3S6. 363. 370. 368. 366. 368. 368. 370. 18976 346. 323. 325. 328. 342. 348. 347. 349. 358. 349. 339. 18977b 354. 338. 334. NO SURVIVING PUPS ON DAY 3 OF LACTATION ---------------------------------------------------------------------------------------------------------------- DAY 14 15 16 17 18 19 20 21 22 ---------------------------------------------------------------------------------------------------------------- 18946b 350. 354. 357. 343. 351. 352. 354. 365. 338. 18950 353. 353. 365. 354. 362. 358. 365. 361. 361. 18951 334. 335. 338. 346. 346. 345. 320. 330. 347. 18952 360. 364. 351. 365. 348. 338. 352. 362. 344. 18953 340. 347. 356. 368. 358. 3S8. 363. 360. 360. 18958 344. 343. 352. 347. 368. 367. 359. 361. 371. 18959 310. 319. 319. 334. 336. 337. 340. 335. 346. 18961 334. 342. 341. 354. 354. 357. 354. 353. 333. 18963 346. 352. 352. 352. 351. 354. 362. 362. 340. 18965 334. 345. 347. 352. 344. 348. 348. 336. 337. 18972 385. 376. 386, 389. 393. 400. 328. 310. 380. 18976 370. 362. 361. 387. 368. 384. 373. 374. 390. 18977b NO SURVIVING PUPS ON DAY 3 OF LACTATION ---------------------------------------------------------------------------------------------------------------- DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. b. Litter was not cross-fostered; values excluded from group averages. PROTOCOL 418-014: OPAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 23 (PAGE 5): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Po GENERATION FEMALE RATS -------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO PHARMACOKINETIC EVALUATION -------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY -------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 1 -------------------------------------------------------------------------------------------------------------- 18910 311. 297. 296. 303. 313. 317. 330. 329. 330. 338. j2 18911 348. 347. 348. 352. 369. 380. 371. 370. 368. 355. 36 18913 320. 323. 328. 324. 348. 354. 336. 345. 339. 342. 35 18915 380. 365. 370. 370. 374. 379. 395. 406. 393. 396. 40 18926 371. 364. 366. 375. 376. 376. 374. 386. 397. 393. 40 18927 312. 309. 322. 322. 331. 330. 333. 337. 344. 348. 33 18939 341. 328. 339. 342. 350. 352. 359. 357. 352. 357. 36 18940 343. 357. 351. 370. 376. 3S3. 358. 365. 372. 373. 36 -------------------------------------------------------------------------------------------------------------- DAY 14 -------------------------------------------------------------------------------------------------------------- 18910 332. 18911 363. 18913 368. 18915 370. 18926 389. 18927 345. 18939 361. 18940 403. -------------------------------------------------------------------------------------------------------------- DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN PATS (SPONSOR'S STUDY NUMBER! T-6295.13) TABLE 23 (PAGE 6): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE PATS --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO PHARMACOKINETIC EVALUATION --------------------------------------------------------------------------------------------------------------- PAT # DOSAGE GROUP 11 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 11 --------------------------------------------------------------------------------------------------------------- 18956 320. 318. 319. 320. 323. 321. 333. 335. 286. 331. 337 18971 311. 311. 309. 312. 311. 311. 317. 315, 316. 318. 320 --------------------------------------------------------------------------------------------------------------- DAY 14 15 16 17 is 19 20 21 22 --------------------------------------------------------------------------------------------------------------- 18956 353. 18971 325. --------------------------------------------------------------------------------------------------------------- DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014! ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBERT T-6295.13) TABLE 24 (PAGE 1): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36- 43a -------------------------------------------------------------------------------------------------------------- 18901 18902 147. 146. 137. 143. 147. b 123. 135. 124. 140. 134. b 18903 18904 c 152. 144. 178. 158. 140. c 131. 128. 128. 125. b 18905 133. 138. 130. c 129. b 18906 144. 149. 150. c 160. b 18907 147. 148. 144. 147. 152. b 18908 116. 129. 126. 136. 122. b 18909 18910 132. 138. 133. 132. 133. b 122, c 130. 162. 135. b 18911 18912 18913 137. 136. 118. 133. 139. 139. 127. 139. iso. 129. 134. 129. 143. 140. 124. b 118. 124. 18914 152. 153. 157. 161. 126. b 18915 147. 148. 153. 104. 188. b 18916 145. 138. 136. 154. 125. 135. 18917 142. 148. 135. 134. 141. b 18918 127. 137. 114. 106. 121. 125. 18919 123. 129. 132. 133. 130. c 18920 136. 134. 126. 127. 128. 128. 18921 130. 128. 132. 132. 129. 119. 18922 129. 154. 146. 138. 148. b 18923 139. 127. 133. 132. c d 18924 159. 160. 165. 162. 159. b 18925 142. 144. 144. 133. 147. b 18926 143. 139. 137. 136. 134. 126. --------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. Rat assigned to cohabitation on day 43 of gestation. C. Spilled feed precluded the calculation of this value. d. value was not recorded. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 24 (PAGE 2): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36- 43a --------------------------------------------------------------------------------------------------------------- 18927 18928 18929 18930 18931 18932 18933 16934 18935 18936 18937 18938 18939 18940 115. 139. 121. 151. 162. 131. 108. 148. 148. 141. 136. 138. 137. 146. 115. 133. 126. 165. 162. 134. 124. 166. 146. 145. 145. 148. 145. iss. 113. 132. 127. 165. 153. 129. 131. 155. 145. 143. c 140. 141. 162. ill. 133. 134. 158. 150. 139. 122. 157. 141. 138. 125. 141. 142. 149. 115. 131. 125. 155. 161. 140. 123. c 134. 136. 120. 141. 142. 146. b b b 150. 151. b b b 134. 80. b 131. b 138, 18941 134. 131. 124. 129. 119. 126. 18942 145. 125. 129. 132. 130. b --------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. Rat assigned to cohabitation on day 42 of study. c. spilled feed precluded the calculation of this value. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN PATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 24 (PAGE 3): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 11 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAYS I- a 8- 15 IS- 22 22- 29 29- 36 36- 43a --------------------------------------------------------------------------------------------------------------- 18943 131. 126. 123. 126. 121. 109. 18944 150. 137. 133. 145. 144. 134. 18945 b 149. b 136. b b 18946 129. 126. 131. 130. 124. 122. 18947 182. 140. 147. b 131. 122. 18948 156. 131. 121. 128. 131. 122. 18949 113. 139. 134. 124. 123. 122. 18950 140. 140. 130. 121. Ill. 106. 18951 124. 121. 124. 125. 118. 101. 18952 145. 144. 137. 135. c 118. 18953 134. 138. 135. 132. 136. 122. 18954 127. 127. 118. 119. 115. 105. 18955 b 135. b b 124. 106. 18956 114. 129. 130. 130. 120. 114. 18957 142. 139. 129. 125. 136. 120. 18958 141. 142. 138. 124. 120. 114. 18959 133. 139. 130. 127. 127. 131. 18960 145. 154. 142. 142. 135. 125. 18961 130. 139. 132. 129. 138. 126. 18962 135. 134. 130. 134. 131. 124. 18963 138. 126. 124. 124. 125. 116. 18964 125. 120. 125. 122. 117. 105. 18965 113. 112. 115. 117. 106. 104. 18966 130. 132. 126. 126. 118. 121. 18967 ISO. 151. 147. 152. 147. 140. 18968 139. 157. 133. 136. 134. 127. --------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. Spilled feed precluded the calculation of this value. c. value appeared incorrectly recorded and was excluded from group averages. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER! T-6295.13) TABLE 24 (PAGE 4): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 11 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAYS I- 8 8- 15 15- 22 22- 29 29- 36 36- 43a --------------------------------------------------------------------------------------------------------------- 18969 151. 151. 149. 151. 141. 142. 18970 132. 133. 129. 129. 125. 119. 18971 148. 148. 134. 132. 124. 117. 18972 139. 159. 139. 142. 137. 130. 18973 137. 135. 130. 129. 125. 113. 18974 161. 146. 118. 124. 120. 121. 18975 152. 156. 154. 153. 156. 140. 18976 130. 129. 137. 130. 141. b 18977 134. 138. 139. 128. 142. b 18978 145. 145. 139. 126. 125. 128. --------------------------------------------------------------------------------------------------------------- DAY - DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. Spilled feed precluded the calculation of this value. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TA13LE 25 (PACE 1): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION F -------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAYS 0 - 1 1 - 2 2 - 3 3 - 4 4 - 5 5 - 6 6 - 7 7 - 8 8 - 9 9 - 10 10 --------------------------------------------------------------------------------------------------------------- 18901 p 18902 NP 18903 P 18904 P 18905 NP 26. 21. 21. 27. 19. 21. 26. 21. 26. 24. 2 23. 20. 21. 26. 26. 24. 25. 22. 23. 26. 1 25. 26. 30. 29. 29. 27. 23. le. 31. 29. 2 20. 21. 22. 22. 25. 24. 25. 20. 21. 23. 2 MATING NOT CONFIRMED 18906 P 18907 P 18908 p 18909 P 18910 p 18911 p 18912 P 18913 P 28. 31. 25. 29. 28. 28. 27. 31. 49. 6. 2 27. 26. 28. 25. 29. 19. 26. 21. 29. 39. 1 25. 22. 21. 22. 20. 26. 20. 28. 79. a 3 25. 23. 23. 21. 25. 24. 26. 25. 25. 24. 2 22. 24. 21. 24. 29. 26. b 23. 21. 18. 2 27. 26. 26. 23. 18. 23. 25. 27. 24. 21. 3 25. 24. 24. 24. 23. 22. 22. 28. 24. 23. 2 22. 24. 23. 20. 18. 20. 23. 20. 21. 23. 2 18914 NP 18. 23. 23. 25. 23. 28. 26. 28. 28. 27. 2 18915 P 25. 28. 27. 26. 30. 30. 29. 27. 29. 25. 3 18916 p 21. 17. 19. 24. 21. 20. 17. 25. 24. 22. 2 18917 P 26. 22. 20. 21. 26. 24. 24. 24. 26. 22. 2 18910 p 22. 19. 22. 20. 28. 26. 28. 24. 28. 27. 2 18919 p 25, 23. 21. b 23. 23. 20. 24. 26. 2S. 2 18920 P 21. 22. 22. 22. 25. 20. 22. 20. 24. 22. 2 18921 NP 18. 8. 17. 18. 16. 16. 14. 17. 17. 14. 1 18922 P 23. 23. 26. 28. 27. 29. 27. 28. 28. 30. 2 18923 P 22. 24. 23. 22. 24. 27. b 19. 22. 23. 2 18924 P 31. 31. 28. 30. 25. 27. 26. 37. 32. 26. 3 18925 NP 17. 23. 23. 22. 14. 25. 21. 22. 12. 23. 2 --------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED DAYS = DAYS OF PRESUMED GESTATION FROM AVERAGES) ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Value could not be calculated. b. Spilled feed precluded the calculation of this value. C. Value was not recorded. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 25 (PAGE 2): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION -------------------------------------------------------------------------------------------------------------- PAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY ------------------------------------------------------------------------------------------------------------ PREGNANCY STATUS DAYS 0 - 1 1 - 2 2 - 3 3 - 4 4 - 5 5 - 6 6 - 7 7 - 8 8 - 9 9 - 10 10 -------------------------------------------------------------------------------------------------------------- 18926 P 18927 P 18928 NP 19. 25. 25. 23. 22. 21. 26. 23. 22. 27. 26. 26. 26. 26. 24. 24. 23. 29. 23. 22. MATING NOT CONFIRMED 18929 P 24. 26. 24. 27, 23. 26. 22. 26. 29. 26. 18930 NP 27. 28. 29. 27. 28. 24. 32. 30. 30. 32. 18931 P 24. 27. 26. 29. 28. 30. 24. 31. 28. 30. 18932 P 19. 24. 24. 25. 22. 23. 25. 20. 28. 26. 18933 P 26. 25. 25. 25. 24. 24. 25. 29. 29. 24. 18934 P 27. 27. 29. 26. 30. 30. 38. 32. 28. 32. 18935 P 14. 18. 22. 28. 27. 26. 21. 19. 25. 24. 18936 P 16. 16. 16. 15. 13. 14. 24. 28. 22. 21. 18937 P 20. 26. 24. 23. 23. 24. 18. 26. 20. 23. 18938 P 23. 28. 25. 25. 27. 25. 27. 26. 25. 28. 18939 P 25. 26. 22. 26. 26. 24. 26. 2S. 26. 25. 18940 P 23. 26. 31. 30. 34. 23. 29. 29. 29. 33. 18941 P 23. 24. 27. 27. 24. 24. 23. 27. 27. 26. 18942 P 24. 23. 22. 24. 27. 24. 25. 24. 25. 24. -------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 25 (PAGE 3): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION I --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG /KG/DAY --------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAYS 13 - 14 14 - 15 15 - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 21 - 22 22 - 23 23 --------------------------------------------------------------------------------------------------------------- 18901 p 18902 NP 18903 P 18904 P 18905 NP 20. 23. 21. 20. 22. 25. 25. 29. 17. 15. 18. 11. 15. 11. 13. 16. 19. 17. 1 29. 26. 20. 28. 26. 30. 25. 26. 21. 30. 25. 19. 27. 21. MATING NOT CONFIRMED 18906 P 28. 15. 30. 31. 31. 28. 39. 18907 P 31. 28. 24. 28. 27. 27. 25. 18908 p 28. 19. 26. 25. 30. 20. 33. 18909 p 32. 25. 33. 31. 16. 28. 21. 22. 3. 18910 p 28. 23. 25. 28. 19. 24. 19. 19. 18911 p 22. 20. 25. 23. 27. 19. 32. 19. 18912 P 23. 20. 25. 29. 31. 21. 35. 18913 P 21. 17. 27. 27. 24. 20. 24. 22. 18914 NP a 19. 18. 23. 15. 19. 13. 18. 11. 22. 1 18915 P 35. 30. 39. 32. 29. 36. 29. 18916 P 18917 P 25. 21. is. 23. 21. 18. 13. 21. 14. 28. 25. 20. 26. 26. 24. 23. 29. 4. 1 18918 p 22. 28. 17. 25. 25. 25. 20. 23. 18919 p 26. 16. 27. 24. 32. 24. 39. 18920 P 24. 22. 18. 26. 27. 30. 21. 18921 NP 12. 17. 16. 9. 11. 16. 16. 11. 16. 23. 18922 P 25. 29. 31. 19. 29. 27. 27. 20, 18923 P 24. 24. 16. 24. 23. 22. 16. 18. 18924 P 28. 17. 27. 26. 33, 20. 36. 18925 NP 19. 25. 20. 12. 14. 18. 17. 16. 14. 16. --------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED DAYS = DAYS OF PRESUMED GESTATION FROM AVERAGES) ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Value was not recorded. PROTOCOL 418-014: OPAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 25 (PAGE 4): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION P --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAYS 13 - 14 14 - 15 15 - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 21 - 22 22 - 23 23 --------------------------------------------------------------------------------------------------------------- 18926 P 18927 P 18928 NP 16. 21. 26. 27. 23. 40. 32, 16. 22. 13. 22. 20. 27. 17. 34. 25. KATING NOT CONFIRMED 18929 P 31. 13. 27. 28. 27. 22. 36. 27. 18930 NP 10. 19. 20. 19. 12. 23. 19. 3. 11. 23. 1 18931 P 27. 14. 29. 27. 31. 20. 32. 18932 P 25. 24. 19. 23. 25. 26. 19. 28. 18933 P 25. 23. 32. 25. 25. 32. 28. 18934 P 27. 31. 27. 19. 26. 28. 27. 18. 18935 P 27. 23. 30. 28. 28. 21. 29. 18936 P 26. 29. 25. 24. 22. 27. 19. 19. 18937 P 20. 12. 21. 22. 24. 19. 28. 25. 18938 P 28. 13. 26. 26. 26. 20. 29. 18939 P 28. 24. 25. 31. 18. 23. 25. 28. 18940 P 17. 20. 22. 23. 17. 28. 26. 7. 18941 P 23. 19. 31. 26. 29. 22. 36. 29. 18942 P 27. 15. 17. 31. 26. 19. 28. 13. ---------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: OPAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 25 (PAGE 5): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION -------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 11 1.6 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAYS 0 - 1 1 - 2 2 - 3 3 - 4 4 - 5 5 - 6 6 - 7 7 - 8 8 - 9 9 - 10 10 -------------------------------------------------------------------------------------------------------------- 18943 P 18944 NP 18945 NP 20. 19. 18. 21. 19. 7. 19. 21. 23. 22. 26. 29. 25. 23. 21. 29. 23. 26. 26. 29. MATING NOT CONFIRMED 18946 P 18947 P 18948 P 18949 NP 18950 P 18951 P 18952 P 18953 P 18954 Np 18955 NP 18956 P 18957 P 18958 P 18959 P 18960 p 18961 P 18962 NP 18963 P 18964 P 18965 P 18966 NP 19. 22. 21. 22. 19. 23. 27. 19. 19, 23. 20. 21. 22. 20. 22. 22. 21. 23. 25. 25. 19. 27. 20. 20. 23. 27. 22. 26. 27. 24. 20. 20. 19. 19. 14. 18. 17. 22. 13. 18. 20. 19. 21. 23. 21. 17. 21. 27, 20. 22. 17. 20. 21. 20. 22. 21. 25. 26. 19. 27. 23. 20. 23. 24. 21. 24. 23. 21. 25. 21. 21. 23. 25. 22. 20. 25. 20. 21. 21. 23. 19. 15. 12. 14. 17. 17. is. 15. 17. 19. 20. 18. 14. 23. 24. 19. 13. 19. 18. 26. 21. 20. 17. is. 15. 20. 22. 23. 19. 21. 17. 23. 21. 21. 22. 22. 20. 26. 25. 23. 23. 22. 22. 23. 21. 20. 21. 20. 25. 26. 17. 20. 20. 18. 22. 19. 22. 23. 21. 24. 22. 22. 23. 23. 19. 22. 21. 25. 27. 21. 19. 22. 22. 24. 22. 23. 23. 21. 20. 27. 13. 18. 18. 12. 13. 17. 17. 15. 11. 18. 17. 20. 22. 22. 23. 22. 22. 23. 26. 25. 20. 21. 19. 20. 23. 20. 23. 24. 19. 23. 21. 22. 17. 21, 18. 17. 26. 18. 20. 24. MATING NOT CONFIRMED 18967 P 14. 24. 23. 25. 20. 18. 20. 30. 24. 27. -------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT DAYS = DAYS OF PRESUMED GESTATION (VALUES EXCLUDED FROM AVERAGES) ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 2S (PAGE 6): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION -------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 11 1.6 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAYS 0 - 1 1 - 2 2 - 3 3 - 4 4 - 5 5 - 6 6 - 7 7 - 8 8 - 9 9 - 10 10 -------------------------------------------------------------------------------------------------------------- 18968 P 18. 22. 18. 22. 19. 23. 25. 24. 24. 21. 18969 P 18970 P 28. 25. 21. 26. 21. 25. 25. 28. 23. 27. 20. 18. 22. 19. a 21. 22. 23. 25. 21. 18971 P 21. 17. 17. 20. 27. 20. 23. 20. 20. 23. 18972 P 20. 18. 22. 21. 23. 24. 23. 25. 24. 25. 18973 P 18. 18. 17. 17. 20. 18. 17. 19. 19. 23. 18974 P 23. 20. 18. 21. 24. 24. 25. 25. 25. 23. 18975 NP 22. 22. 22. 19. 19. 23. 23. 22. 24. 26. 18976 P 18. 20. 20. 24. 23. 23. 27. 22. 31. 20. 18977 P 19. 20. 21. 23. 24. 25. 26. 26. 22. 25. 18978 NP 17. 16. 20. 20. 20. 25. 21. 24. 20. 19. -------------------------------------------------------------------------------------------------------------- P - PREGNANT NP = NOT PREGNANT (VALUES DAYS = DAYS OF PRESUMED GESTATION EXCLUDED FROM AVERAGES) ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Spilled feed precluded the calculation of this value. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 25 (PAGE 7): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FE ---------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 11 1.6 MG/KG/DAY ---------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAYS 13 - 14 14 - 15 15 - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 21 - 22 22 - 23 23 ---------------------------------------------------------------------------------------------------------------- 18943 P 18944 NP 18945 NP 18946 P 18947 P 18948 P 18949 NP 18950 P 18951 P 18952 P 18953 P 18954 NP 18955 NP 18956 P 18957 P 18958 P 18959 p 18960 P 18961 p 18962 NP 18963 P 18964 P 18965 P 18966 NP 20. 24. 31. 30. 27. 26. 33. 18. S. 17. 15. 11. MATING NOT CONFIRMED 8. 21. is. 7. 12. 21 18. is. 26. 26. 23. 28. 32. 7. 29. 22. 32. 30. 25. 13. 23. 14. 32. 32. 36, 21. 39, 19. 9. 16. 9. 17. 12. 17. 9. 7. 18. 16 16. 22. 26. 30. 20. 35. 29. IS. 25. 26. 29. 22. 37. 28. 24. 26. 13. 26. 24. 23. 20. 24. 16. 26. 25. 28. 23. 34. 16. 15. 18. 7. 12. 16. 14. 10. 15. 14. 5 20. 14. 18. 10. is. 13. 18. 13. 18. 15. 7 16. 19. 21. 27. 23. 34. 30. 14, is. 25. 27. 17. 31. 27. 27. 20. 20. 26. 25. 27. 25. 38. 33. 13. 24. 23. 29. 15. 23. 29. 27. 19. 23. 14. 23. 25. 27. 24. 30. 19. 21. 24. 19. 28. 27. 24. 24. 17. 19. 18. S. 16. 15. 15. B. 18. 19. 8 28. 28. 19. 26. 27. 20. 15. 20. 25. 24. 22. 29. 28. 24. 23. 15. 26. 26. 30. 21. 34. 25. MATING NOT CONFIRMED 18967 P 28. 10. 27. 29. 29. 27. 39. ---------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING SI'IJDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 25 (PACE 8): MATERNAL FEED CONSUMPTION VAI,UES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION F --------------------------------------------------------------------------------------------------------------- PAT # DOSAGE GROUP 11 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAYS 13 - 14 14 - 15 15 - 16 16 - 17 17 - IS 18 - 19 19 - 20 20 - 21 21 - 22 22 - 23 23 --------------------------------------------------------------------------------------------------------------- 18968 P 23. 28. 29. 24. 29. 29. 24. 20. 18969 P 23. 20. 27. 31. 31. 21. 42. 18970 P 20. 21. 17. 24. 22. 26. 20. 10971 P 21. 14. 19. 20. 20. 18. 25. 21. 9. 18972 P 28. 18. 29. 27. 31. 25. 37. 18973 P 24. 15. 22. 21. 27. 22. 26. 18974 P 30. 29. 18. 30. 26. 30. 23. 18975 NP 18976 P 15. 10. 20. 16. 11. 14. 24. 14. 27. 24. 21. 25. 24. 23. 18. 9. 15. 2 18977 P 26. 31. 31. 19. 21. 28. 29. 28. 18978 NP 16, 26. 18. 3. 22. 21. 19. 6. 27. 22. --------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT DAYS = DAYS OF PRESUMED GESTATION (VALUES EXCLUDED FROM AVERAGES) ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 26 (PAGE 1): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RA -------------------------------------------------------------------------------------------------------------- PATS ASSIGNED TO CROSS-FOSTERING -------------------------------------------------------------------------------------------------------------- DAYS 1 - 4 4 - 7 7 - 10 10 - 14a -------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP I b 0 (VEHICLE) MG/KG/DAY SUBSET A -------------------------------------------------------------------------------------------------------------- 18903 47. 109. 130. 232. 18907 51. 117. 135. 235. 18908 48. 104. 138. 243. 18909 48. 121. 149. 207. 18918C 82. 145. 156. 264. 18920 63. 124. 116.d 226. 18924 48. 124. 134. 218. 18929 69. 127. 147. 257. 18934 49. 108. 114. 215. 18936c 46. 117. 148. 183. 18937 62. 123. 130. 250. 18941 61. 104. 132. 219. 18942 85. 120. 140. 239. -------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP I e 0 (VEHICLE) MG/KG/DAY SUBSET B -------------------------------------------------------------------------------------------------------------- 18901 18904 18906 18912 18917 18919 18922 18923 18931 96. 135. 174. 255. 71. 143. 160.d 200. 90. 128. 168. 248. 64. 122. 154. 246. 80. 134. 147. 240. 74. 130. 147. 227. 62. 117. 145. 227. 82. 149. 177. 242. 72. 130. 148. 244. 18932 1893S 74. 118. 145. 203. 60. 110. 133. 218. 18938 70. 128. 156. 243. -------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. it is presumed that the pups begin to consume the maternal feed after day 14 of lactation. b. vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. C. Litter was not cross-fostered; values excluded from group averages. d. Water access was interrupted; values excluded from group averages. e. vehicle control group dams cross-fostered with vehicle control litters. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN PATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 26 (PAGE 2): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS ----------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING ----------------------------------------------------------------------------------------------------------------- DAYS 1 - 4 4 - 7 7 - 10 10 - 14a ----------------------------------------------------------------------------------------------------------------- PAT # DOSAGE GROUP II b 1.6 MG/KG/DAY SUBSET C ----------------------------------------------------------------------------------------------------------------- 18943 18947 10948 18957 18960 18964 18967 18968 73. 123. 141. 240. 68. 113. 126. 211. 48. 95. 106. 193. 51. 112. 128. 208. 46. 102. 116. 191.c 54. 98. 126. 182. 68. 112. 138. 243. 50. 108. 125. 197. 18969 63. 116. 128. 231. 18970 54. 109. 114. 191. 18973 45. 51. 113. 182. 18974 31. NO SURVIVING PUPS ON DAY 4 OF LACTATION ----------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP II d 1.6 MG/KG/DAY SUBSET D ----------------------------------------------------------------------------------------------------------------- 18946e 189so 18951 29. 76. 77. 124. 68. 121. 135. 227. 67. 99. 115. 205. 18952 18953 18958 18959 74. 140. 138. 215. 59. 104. 127. 203. 51. 87. 119. 202. 46. 104. 135. 179. 18961 18963 64. 119. 134. 201. 48. 114. 127. 233. 18965 18972 82. 116. 133. 209. 77. 140. 148. 258. 18976 66. 127. 118. 213. 18977e NO SURVIVING PUPS ON DAY 3 OF LACTATION ----------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. It is presumed that the pups begin to consume the maternal feed after day 14 of lactation. b. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. C. Water access was interrupted; value excluded from group averages. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. e. Litters were not cross-fostered; values excluded from group averages. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NLTMBER: T-6295.13) TABLE 26 (PAGE 3): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RAI -------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO PHARMACOKINETIC EVALUATION -------------------------------------------------------------------------------------------------------------- DAYS 1 - 4 4 - 7 7 - 10 10 - 14 -------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 18910 61. 73. 114. 145. 18911 107. 149. 148. 251. 18913 68. 134. 140. 261. 18915 se. 138. 216. 246. 18926 97. 131. 181. 300. 18927 66. 119. 137. 231. 18939 86. 120. 154. 230. 18940 82. 106. 141. 26S. -------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 11 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 18956 73. 97. 122. 258. 18971 31. 44. 57. 77. --------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 27 (PAGE 1): NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY Fo GENERATION FEMALE PATS/Fl GENERATION LITTERS AND SEX - INDIVIDUAL DATA - -------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING -------------------------------------------------------------------------------------------------------------- LITTER DELIVERED TOTAL PUPS NUMBER OF LIVE FOSTER PU PAT/ LIVE STILL- TOTAL IMPLAN- CROSS- COMPLETION OF DAY POSTPA LITTER BORN BORN BORN TATIJDNS FOSTER FOSTERED 1 4& 4b 7 NUMBER N N N N LITTER N m F m p N F m p -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP I c 0 (VEHICLE) MG/KG/DAY SUBSET A -------------------------------------------------------------------------------------------------------------- 18903 18 0 18 le 18976 15 7 8 6 7 5 5 5 5 18907 19 0 19 20 18963 13 5 8 5 7 5 5 5 5 18906 16 0 16 17 189S3 16 4 12 3 12 3 7 3 7 18909 16 0 16 17 18961 16 a 8 7 8 5 5 5 5 18918d 15 0 is 16 18946 17 9 a 7 8 5 5 5 5 18920 17 0 17 17 18952 15 8 7 8 5 5 5 5 5 18924 16 0 16 17 18972 17 9 a 8 7 5 5 5 5 18929 18 0 18 18 18950 16 a 8 7 8 5 5 5 5 18934 16 0 16 le 18959 14 7 7 7 6 5 5 5 5 18936d 12(l) 0 12 15 18977 12 7 5 7 4 6 4 6 4 18937 14 1 15 18 l89s8 13 5 8 5 8 5 5 5 5 18941 14 0 14 17 18951 13 8 5 5 4 5 4 5 3 18942 17 0 17 17 18965 18 9 9 9 9 5 5 5 5 -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP I e 0 (VEHICLE) MG/KG/DAY SUBSET B -------------------------------------------------------------------------------------------------------------- 18901 16 0 16 17 18923 16 6 10 6 10 5 5 5 5 18904 is 3 18 20 18922 15 7 8 7 8 5 5 5 5 18906 16 0 16 19 18935 19 8 11 8 10 5 5 5 5 18912 is 1 16 16 18938 20 11 9 11 8 5 5 5 5 18917 14 0 14 1'7 18932 16 9 7 9 7 5 5 5 5 18919 17 0 17 18 18931 12 7 5 7 5 5 5 5 5 18922 15 0 15 18 18904 15 5 10 5 10 5 5 5 5 18923 17(l) 0 17 17 18901 16 6 10 6 9 5 5 5 5 18931 12 0 12 12 18919 17 7 10 7 10 5 5 5 5 18932 16 0 16 is 18917 14 6 8 6 8 5 5 5 5 18935 19 0 19 21 18906 16 7 9 7 9 5 5 5 5 18938 20 0 20 20 18912 15 13 2 13 2 a 2 8 2 --------------------------------------------------------------------------------------------------------------- M MALE F = FEMALE ( = NUMBER OF PUPS DYING PRIOR TO CROSS-FOSTERING ON DAY 1 POSTPARTUM a. Number of live pups preculling. b. Number of live pups postculling. c. Vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. Litter were not cross-fostered; values excluded from group averages. e. vehicle control group dams cross-fostered with vehicle control litters. PROTOCOL 418-014: OPAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 27 (PAGE 2): NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY Fo GENERATION FEMALE RATS/Fl GENERATION LITTERS AND SEX - INDIVIDUAL DATA - -------------------------------------------------- ---------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING -------------------------------------------------------------------------------------------------------------- LITTER DELIVERED TOTI%L PUPS NUMBER OF LIVE FOSTER PU RAT/ LIVE STILL- TOTAL IMPLAN- CROSS- COMPLETION OF DAY POSTPAI LITTER BORN BORN BORN TATibNs FOSTER FOSTERED 1 4a 4b 7 NUMBER N N N N' LITTER N m F m p N F m p ---------------------------------------- --------------------------------------------------------------------- MATERNAL DOSAGE GROUP II C 1.6 MG/KG/DAY SUBSET C -------------------------------------------------------------------------------------------------------------- 18943 16 0 16 16 18967 15 5 10 5 10 s 5 5 5 18947 15 0 15 16 18974 14 8 6 a 6 s 5 5 5 18948 17 0 17 19 18969 17 11 6 6 2 6 2 6 2 18957 15 0 15 15 18970 13 4 9 3 8 3 7 3 7 18960 15 0 is 17 18973 15 5 10 3 8 3 7 3 7 18964 10 0 10 13 18968 15 8 7 7 7 5 s 5 5 18967 15 0 15 17 18943 16 9 7 9 7 s 5 5 5 18968 15 0 15 16 18964 10 2 8 1 a 1 8 1 8 18969 17 0 17 19 18948 17 12 5 12 5 5 5 5 5 18970 13 0 13 13 18957 15 9 6 9 5 s 5 5 5 18973 18974 16(l) 0 16 14 0 14 16 18960 17 18947 15 10 5 10 4 6 4 64 is 6 9 -- - - - - -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP II d 1.6 MG/KG/DAY SUBSET D -------------------------------------------------------------------------------------------------------------- 18946e 17 0 17 17 18918 15 9 a 3 1 3 1 2 1 18950 16 0 16 17 18929 18 8 10 8 10 5 5 5 5 18951 13 0 13 13 18941 14 8 6 8 6 5 5 5 5 18952 15 2 17 17 18920 17 8 9 8 9 5 5 5 5 18953 16 0 16 18 18908 16 8 8 8 8 5 5 5 5 18958 13 0 13 14 18937 14 6 8 6 8 5 5 5 5 18959 14 0 14 15 18934 16 9 7 9 6 5 5 5 5 18961 16 0 16 16 18909 16 5 11 5 11 5 5 5 5 18963 13 0 13 13 18907 19 11 8 11 8 5 5 55 18965 18 0 18 18 18942 17 6 11 6 11 5 s 55 18972 17 0 17 18 18924 16 7 9 7 9 5 5 5 5 18976 15 0 15 15 18903 18 8 10 7 10 5 s 5 5 18977e 12 0 12 16 18936 12 7 4 -- - - - - -------------------------------------------------------------------------------------------------------------- M MALE F = FEMALE ( = NUMBER OF PUPS DYING PRIOR TO CROSS-FOSTERING ON DAY 1 POSTPARTUM a. Number of live pups preculling. b. Number of live pups postculling. C. 1.6 mg/kg/day dosage group dams cross-foatered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. e. Litter were not cross-fostered; values excluded from group averages. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 27 (PAGE 3): NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA Fo GENERATION FEMALE RATS/Fl GENERATION LITTERS -------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO PHARMACOKINETIC EVALUATION -------------------------------------------------------------------------------------------------------------- LITTER DELIVERED NUMBER OF LIVE PUPS AT RAT/ LIVE STILL- TOTAL COMPLETION OF DAY POSTPARTUM LITTER BORN BORN BORN 1 4 7 14 NUMBER N N N mF mF mF mF -------------------------------------------------------------------------------------------------------------- IMPLANTATIONS N TOTAL MATERNAL DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 18910 5 0 5 32 32 32 32 8 18911 16 0 16 11 5 11 5 11 4 11 4 16 18913 18 0 18 16 2 15 2 14 2 14 2 19 18915 is 2 17 4 11 4 11 4 11 4 11 17 18926 12 0 12 66 66 66 66 13 18927 17 0 17 89 88 88 87 17 18939 is 0 is 78 78 78 78 15 18940 16 0 16 10 6 10 6 10 6 10 6 18 -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP 11 1.6 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 18956 12 1 13 57 57 57 57 16 18971 1 2 3 -1 -1 -1 -1 3 -------------------------------------------------------------------------------------------------------------- M = MALE F = FEMALE PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 28 (PAGE 1): DURATION OF GESTATION AND AVERAGE DURATION OF DELIVERY Fo GENERATION FEMALE PATS (MINUTES) PER PUP PER LITTER - IN -------------------------------------------------------------------------------------------------------------- PATS ASSIGNED TO CROSS-FOSTERING -------------------------------------------------------------------------------------------------------------- RAT/ DURATION OF GESTATION DURATION OF GESTATION AVERAGE DURATION OF LITTER (DAYS) a (WHOLE DAYS) DELIVERY PER PUP NUMBER N N (MINUTES) b -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP I c 0 (VEHICLE) MG/KG/DAY SUBSET A -------------------------------------------------------------------------------------------------------------- 18903 21.6 22 16.9 18907 21.5 22 14.5 18908 21.7 22 8.5 18909 22.5 23 15.5 18918d 22.6 23 7.2 18920 21.6 22 9.1 18924 21.6 22 6.5 18929 22.4 23 6.7 18934 21.5 22 5.1 18936d 21.7 22 25.1 18937 22.5 23 23.2 18941 22.4 23 IS.3 18942 21.8 22 9.5 -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP I e 0 (VEHICLE) MG/KG/DAY SUBSET B -------------------------------------------------------------------------------------------------------------- 18901 18904 22.3 22.6 23 21.0 23 11.4 18906 18912 21.6 21.6 22 8.8 22 12.7 18917 22.5 23 13.4 10919 21.7 22 9.1 18922 21.6 22 13.3 18923 22.8 23 11.7f 18931 21.6 22 12.4 18932 22.5 23 7.2 18935 21.6 22 7.4 18938 21.6 22 6.4 --------------------------------------------------------------------------------------------------------------- a. Calculated to the nearest tenth of a day. b. Calculated as the time (in minutes) elapsed between delivery of the first and last pup, divided by n-I pul C. vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. Vehicle control group dams cross-fostered with vehicle control litters. e. Litters were not cross-fostered; values excluded from group averages. f. Partuition period only partially observed. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 28 (PAGE 2): DURATION OF GESTATION AND AVERAGE DURATION OF DELIVERY (MINUTES) PER PUP PER LITTER - IN Fo GENERATION FEMALE PATS --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING -------------------------------------------------------------------------------------------------------------- RAT/ DURATION OF GESTATION DURATION OF GESTATION AVERAGE DURATION OF LITTER (DAYS) a (WHOLE DAYS) DELIVERY PER PUP NUMBER N N (MINUTES) b -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP II c 1.6 MG/KG/DAY SUBSET C --------------------------------------------------------------------------------------------------------------- 18943 18947 21.5 21.5 22 4.9 22 10.8 18948 21.6 22 11.2 18957 21.6 22 11.1 18960 21.6 22 3.1 18964 21.8 22 9.9 18967 21.5 22 4.6 18968 21.7 22 9.5 18969 18970 21.6 21.7 22 11.2 22 8.3 18973 18974 21.5 21.5 22 11.0 22 8.6 --------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP Ii d 1.6 MG/KG/DAY SUBSET D --------------------------------------------------------------------------------------------------------------- 18946e 21.5 22 12.3f 18950 21.5 22 8.3 18951 21.6 22 11.2 18952 21.5 22 13.2 18953 21.7 22 14.1 18958 21.7 22 5.5 18959 21.5 22 9.2 18961 21.5 22 7.9 18963 21.6 22 11.2 18965 21.4 22 6.4 18972 21.7 22 4.4 18976 21.7 22 13.8 18977e 21.6 22 24.4 --------------------------------------------------------------------------------------------------------------- a. Calculated to the nearest tenth of a day. b. Calculated as the time (in minutes) elapsed between delivery of the first and last pup, divided by n-1 pup c. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dame cross-fostered with vehicle control group litters. e. Litters were not cross-fostered; values excluded from group averages. f. Partuition period only partially observed. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 28 (PAGE 3): DURATION OF GESTATION AND AVERAGE DURATION OF DELIVERY (MINUTES) PER PUP PER LITTER - IND Fo GENERATION FEMALE PATS --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO PHARMACOKINETIC EVALUATION --------------------------------------------------------------------------------------------------------------- PAT/ DURATION OF GESTATION DURATION OF GESTATION AVERAGE DURATION OF LITTER (DAYS) a (WHOLE DAYS) DELIVERY PER PUP @MBER N N (MINUTES) b --------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 18910 21.7 22 14.8 18911 21.8 22 14.9 18913 22.4 23 6.1 18915 21.6 22 30.4 18926 22.4 23 5.2 18927 22.3 23 9.5 18939 21.9 22 9.6 18940 21.7 22 9.2d --------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP 11 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 18956 22.5 23 8.2 18971 c 24 c --------------------------------------------------------------------------------------------------------------- a. Calculated to the nearest tenth of a day. b. Calculated as the time (in minutes) elapsed between delivery of the first and last pup, divided by n-1 pup C. Partuition period not observed. d. Partuition period only partially observed. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 29 (PAGE 1); PUP BODY WEIGHT LITTER AVERAGES FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GEN ---------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING ---------------------------------------------------------------------------------------------------------------- PAT/ FOSTER DAY 1 DAY 4a DAY 4b DAY 7 DAY 14 LITTER m F T m F T m F T m F T m F T m ---------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP I c 0 (VEHICLE) MG/KG/DAY SUBSET A ---------------------------------------------------------------------------------------------------------------- 18903 5.6 5.2 5.4 5.9 5.9 5.9 6.1 5.9 6.0 10.5 9.6 10.0 24.7 23.2 24.0 41. 18907 6.1 S.8 5.9 7.8 7.4 7.6 7.8 7.6 7.7 13.0 12.6 12.8 27.9 27.1 27.5 44. 18908 5.8 5.4 5.5 7.1 7.0 7.0 7.1 7.2 7.2 12.6 11.7 11.9 26.8 26.2 26.4 42. 18909 5.5 5.5 5.5 7.4 7.2 7.3 7.7 7.7 7.7 13.4 13.4 13.4 28.1 27.8 28.0 42. 18918d 5.6 5.1 5.3 10.0 9.2 9.6 10.0 9.1 9.6 16.5 15.0 15.8 33.0 30.6 31.8 50. 18920 5.3 5.2 5.3 6.7 7.0 6.8 7.3 7.0 7.1 12.3 11.8 12.0 25.0 24.5 24.7 41. 18924 5.7 5.5 5.6 7.5 6.7 7.1 7.5 7.3 7.4 12.5 12.0 12.3 25.9 25.3 25.6 41. 18929 5.6 5.3 5.4 7.6 7.2 7.4 8.1 7.4 7.8 13.7 12.5 13.1 30.2 27.9 29.0 47. 18934 5.8 5.4 5.6 7.4 7.0 7.2 7.6 7.0 7.3 13.0 12.4 12.7 27.6 26.3 27.0 42. 18936d 6.2 5.8 6.1 8.7 7.7 8.3 8.6 7.7 8.2 13.1 12.0 12.7 25.1 23.0 24.2 39. 18937 6.0 5.5 5.7 6.0 7.3 7.6 8.0 7.5 7.7 11.9 11.5 11.7 24.9 25.2 2S.1 40. 18941 6.2 5.9 6.1 8.2 6.9 7.6 8.2 6.9 7.6 12.8 12.7 12.8 28.2 29.2 28.6 44. 18942 5.3 5.1 5.2 7.0 7.0 7.0 7.2 7.3 7.3 12.4 12.5 12.4 28.0 28.4 28.2 41. ---------------------------------------------------------------------------------------------------------------- 14ATERNAL DOSAGE GROUP I e 0 (VEHICLE) MG/KG/DAY SUBSET B ---------------------------------------------------------------------------------------------------------------- 18901 6.7 6.2 6.4 9.4 8.6 8.9 9.5 8.5 9.0 15.6 14.3 15.0 31.7 30.1 30.9 50. 18904 5.9 5.6 5.7 7.6 7.0 7.3 8.0 7.4 7.7 14.2 12.6 13.4 28.9 26.5 27.7 44. 18906 5.8 5.5 5.6 8.2 7.6 7.9 8.4 8.1 8.3 13.5 13.4 13.4 28.7 28.3 28.5 49. 18912 5.5 5.2 5.4 6.8 6.2 6.6 7.0 6.3 6.6 12.5' 12.7 12.6 29.8 27.5 28.6 48. 18917 7.0 6.7 6.9 8@7 8.4 8.5 8.6 8.4 8.5 13.8 13.6 13.7 28.9 28.4 2B.6 45. 18919 6.4 6.3 6.4 9.5 9.3 9.4 9.3 9.3 9.3 15.6 1S.1 15.4 30.9 29.9 30.4 49. 18922 6.8 6.4 6.5 8.9 8.5 8.6 8.9 8.9 8.9 15.5 15.1 15.3 30.8 31.0 30.9 48. 18923 7.0 6.7 6.8 9.6 8.9 9.2 9.8 8.8 9.3 16.1 14.3 15.2 30.5 28.1 29.3 so. 18931 6.4 6.0 6.2 8.3 8.0 8.1 8.2 8.2 8.2 13.8 13.8 13.8 30.6 30.5 30.6 48. 18932 6.8 6.2 6.4 9.1 8.6 8.8 9.2 8.7 8.9 14.5 13.9 14.2 27.4 26.9 27.2 43. 18935 6.3 5.8 6.0 8.4 7.7 8.0 8.5 8.1 8.3 13.5 13.0 13.2 26.2 25.9 26.0 42. 18938 6.1 5.8 6.1 8.5 8.4 8.5 8.7 8.4 8.6 13.8 13.8 13.8 29.2 29.0 29.1 44. ---------------------------------------------------------------------------------------------------------------- M = MALE F = FEMALE ALL WEIGHTS WERE RECORDED a. Preculling T = TOTAL (SUM OF PUP WEIGHTS/NUMBER OF LIVE PUPS) DAY = DAY POSTPARTUM IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. b. Postculling C. Vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. Litters were not cross-fostered; values excluded from group averages. e. Vehicle control group dams cross-fostered with vehicle control litters. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 29 (PAGE 2): PUP BODY WEIGHT LITTER AVERAGES FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GE --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- RAT/ POSTER LITTER DAY I m F T DAY 4a m F T DAY 4b m F T DAY 7 m F T DAY 14 m F T m ------------------------------------------------------------------------------------------------------------ 7-- MATERNAL DOSAGE GROUP Il c 1.6 MG/KG/DAY SUBSET C --------------------------------------------------------------------------------------------------------------- 18943 18947 18948 18957 18960 18964 18967 18968 18969 18970 18973 18974 5.8 5.6 5.7 8.0 8.1 8.0 8.0 8.2 8.1 12.9 13.0 13.0 28.4 28.1 28.2 43 5.8 5.6 5.7 7.6 7.1 7.4 7.8 7.2 7.5 12.5 11.7 12.1 26.4 24.2 25.3 43 5.8 5.4 5.7 6.3 5.8 6.2 6.3 5.8 6.2 10.1 9.0 9.8 23.7 20.8 23.0 36 6.2 5.7 5.9 7.6 7.4 7.5 7.6 7.5 7.6 12.3 12.3 12.3 24.9 25.4 25.2 39 5.4 5.3 S.4 6.8 6.2 6.3 6.8 6.4 6.5 10.3 9.3 9.6 22.9 20.4 21.2 35 6.1 6.0 6.0 6.6 6.7 6.6 6.6 6.9 6.8 10.3 11.0 10.7 21.4 22.3 21.8 33 5.8 5.5 5.7 8.0 7.2 7.6 8.3 7.0 7.7 14.0 11.3 12.6 29.5 24.9 27.2 47 6.6 6.2 6.3 10.4 8.7 8.9 10.4 8.7 8.9 15.4 13.4 13.6 30.4 26.6 27.1 50 5.6 S.6 5.6 7.4 7.5 7.4 7.7 7.5 7.6 12.7 11.9 12.3 27.0 2S.6 26.3 40 5.7 5.4 5.6 7.1 5.6 6.6 7.4 S.6 6.5 12.6 9.4 11.0 25.1 20.0 22.5 39 5.6 5.4 5.6 7@4 7.2 7.3 7.5 7.2 7.4 11.4 10.9 11.2 23.0 22.9 23.0 36 S.7 5.3 5.4 NO SURVIVING PUPS --------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP II d 1.6 MG/KG/DAY SUBSET D --------------------------------------------------------------------------------------------------------------- 18946e 18950 18951 18952 18953 18958 18959 18961 18963 18965 18972 18976 18977e 6.9 6.4 6.7 7.1 6.3 6.7 7.8 7.5 7.6 6.4 5.9 6.1 7.0 6.5 6.8 7.2 6.4 6.7 6.0 S.4 S.0 6.3 6.1 6.2 5.8 5.4 5.6 6.0 5.7 5.8 6.4 5.8 6.0 6.2 5.9 6.0 6.7 6.1 6.5 6.3 5.2 6.0 6.3 5.2 6.0 12.4 7.8 10.8 30.3 19.3 26.6 48 8.1 7.3 7.7 8.4 7.6 8.0 13.3 12.3 12.8 26.9 25.4 26.1 44 8.8 8.4 8.6 9.0 8.4 8.7 12.6 11.2 11.9 24.2 21.6 22.9 39 8.2 7.4 7.8 8.7 7.6 8.2 14.8 13.3 14.0 30.7 28.1 29.4 47 8.2 8.2 8.2 8.6 8.5 8.6 12.7 11.9 12.3 25.4 24.8 25.1 41 9.2 8.1 8.6 9.1 8.4 8.7 12.9 12.3 12.6 24.7 24.0 24.4 40 8.2 7.6 8.0 8.0 7.7 7.8 13.0 12.5 12.7 27.1 26.2 26.6 42 8.3 7.7 7.9 8.3 8.5 8.4 13.3 13.1 13.2 26.3 25.7 26.0 42 7.1 6.4 6.8 7.3 6.6 6.9 12.2 11.7 12.0 26.8 25.9 26.4 43 8.1 7.6 7.8 8.2 7.7 8.0 12.7 12.3 12.5 25.6 24.9 25.2 44 9.0 8.1 B.S 9.0 8.3 8.7 14.8 14.0 14.4 31.4 29.9 30.6 46 7.4 7.1 7.2 7.6 7.2 7.4 12.8 12.5 12.6 26.2 24.8 25.5 41. NO SURVIVING PUPS ---------------------------------------------------------------------------------------------------------------- M= ALL a. b. MALE F= WEIGHTS WERE Preculling Postculling FEMALE RECORDED T = TOTAL (SUM IN GRAMS (G). OF PUP WEIGHTS/NUMBER MEAN LITTER WEIGHTS OF LIVE INCLUDE PUPS) ONLY DAY = DAY POSTPARTUM WEIGHTS OF LIVE PUPS. C. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. e. Litters were not cross-fostered; values excluded from group averages. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN PATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 29 (PAGE 3): PUP BODY WEIGHT LITTER AVERAGES FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GEN ---------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO PHARMACOKINETIC EVALUATION ---------------------------------------------------------------------------------------------------------------- PAT/ LITTER DAY 1 DAY 4 DAY 7 DAY 14 NUMBER m F T m F T m F T m F T ---------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY ---------------------------------------------------------------------------------------------------------------- 18910 7.4 7.1 7.3 a a a 17.S 16.4 17.1 33.8 32.1 33.1 18911 6.6 6.5 6.6 8.9 8.4 8.8 12.4 12.S 12.4 23.7 24.1 23.8 18913 6.2 5.7 6.2 8.0 7.6 8.0 10.9 10.6 10.9 21.2 19.6 21.0 1891s 6.4 6.1 6.2 a a a 12.6 13.0 12.9 24.5 2S.3 2S.0 18926 7.0 7.0 7.0 11.9 12.4 12.2 15.9 15.6 15.8 30.0 29.4 29.7 18927 6.6 6.1 6.3 8.8 8.2 8.5 12.3 10.8 ll.S 22.4 20.6 21.6 18939 6.4 6.0 6.2 a a a 12.9 12.5 12.7 23.6 22.6 23.1 18940 6.2 5.8 6.1 8.5 8.3 8.4 10.8 10.8 10.8 19.8 18.8 19.4 ---------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP 11 1.6 MG/KG/DAY ---------------------------------------------------------------------------------------------------------------- 189S6 6.4 6.0 6.2 9.3 B.8 9.0 13.1 12.1 12.5 24.2 23.4 23.8 18971 --- 7.6 7.6 --- 9.1 9.1 --- 9.2 9.2 --- 11.7 11.7 ---------------------------------------------------------------------------------------------------------------- M = MALE ALL WEIGHTS a. Values F = FEMALE T = TOTAL WERE RECORDED IN GRAMS were not recorded. (SLqi OF (G).,@@ PUP WEIGHTS/NUMBER MEAN LITTER WEIGHTS OF LIVE INCLUDE PUPS) ONLY DAY WEIGHTS = DAY POSTPARTUM OF LIVE PUPS. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 1)! PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 is 16 1 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP I a 0 (VEHICLE) MG/KG/DAY SUBSET A POSTPARTUM DAY ---------------------------------------------------------------------------------------------------------------- 18903 18907 5.6 6.1 5.3 5.5 5.6 5.3 5.7 4.9 5.0 5.4 5.4 5.5 5.6 5.2 5.0 6.2 6.0 5.8 6.1 6.3 5.9 5.8 5.7 5.8 5.9 S.8 5.5 6.2 18908 18909 18918b 18920 16924 18929 18934 6.0 5.9 6.0 5.4 5.8 5.2 5.1 5.5 5.5 5.8 4.9 5.4 5.1 5.4 5.3 5.8 5.2 5.7 5.4 5.2 6.0 5.3 5.8 5.6 5.5 5.2 5.5 5.2 5.6 5.6 5.7 5.4 S.2 5.3 5.5 5.5 5.7 5.4 5.5 5.3 6.7 5.5 5.1 5.0 5.1 5.0 5.0 4.9 5 5.6 5.1 5.3 5.2 5.3 5.6 5.2 5.4 5.2 5.0 5.5 5.4 5.0 5.1 5.2 5.6 S.3 5.8 5.9 5.7 6.0 5.9 5.8 5.0 5.5 5.7 5.4 5.5 5.4 5.0 5.7 5 5.6 6.1 5.6 5.5 5.5 5.6 5.5 5.6 5.4 5.5 4.9 5.0 5.3 5.5 5.3 5.2 5.6 5.7 5.6 6.0 5.8 6.2 5.7 5.3 5.6 5.3 5.5 5.3 5.4 5.5 18936b 6.4 6.3 5.9 6.6 6.0 6.2 6.4 S.9 5.7 5.9 5.9 5.8 18937 6.2 5.9 6.0 6.2 5.8 5.6 5.1 5.5 5.7 5.5 S.7 5.6 5.6 18941 6.3 5.9 6.3 6.4 6.8 6.3 6.0 5.5 5.6 6.0 5.7 6.6 5.8 18942 S.3 5.2 5.1 4.9 5.3 5.5 5.6 5.5 5.6 5.5 5.0 5.0 4.9 4.4 5.3 5.3 5 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY SUBSET B POSTPARTUM DAY --------------------------------------------------------------------------------------------------------------- 18901 18904 18906 18912 18917 18919 6.4 6.7 6.7 6.9 7.1 6.2 6.3 6.7 6.5 6.4 6.6 6.2 5.4 5.8 5.8 6.2 5.2 6.4 6.5 6.1 5.6 6.0 5.5 5.3 5.9 6.1 5.3 5.7 5.4 6.1 S.1 6.1 5.8 5.7 6.1 5.0 5.9 6.1 5.6 5.6 5.4 4.9 S.7 5.1 5.7 5.5 5.5 5 5.7 5.2 5.9 5.0 5.2 5.6 5.6 5.4 5.6 5.6 5.3 5.1 5.6 5.1 S.5 5.2 4 7.2 6.9 6.7 6.9 7.3 6.5 7.5 6.5 7.3 6.8 6.6 7.0 6.9 6.5 6.9 6.4 6.0 6.3 6.4 6.2 6.9 6.5 6.5 6.4 6.2 6.3 6.4 6.1 18922 18923 18931 18932 7.3 6.7 6.8 5.9 7.4 6.3 6.7 7.0 5.7 5.7 6.0 6.9 6.4 6.5 6.7 7.3 6.7 6.9 7.0 7.2 6.9 6.5 6.3 6.4 6.7 7.2 6.3 6.8 6.9 6.9 6.8 6.4 6.3 6.7 6.3 6.7 6.6 6.1 6.4 6.2 6@2 6.1 5.9 6.0 6.3 5.3 5.9 6 6.8 6.8 6.3 7.0 7.0 6.9 6.0 5.7 6.5 6.5 6.1 6.1 6.3 6.3 18935 6.3 6.3 6.2 6.3 6.5 5.9 6.6 5.7 5.7 6.0 5.5 6.4 6.1 4.7 5.8 6.0 18938 5.9 6.1 6.3 6.1 6.3 6.3 5.9 6.2 6.4 6.2 6.1 5.8 6.2 5.9 5.8 ---------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST LETTER M-MALE, F-FEMALE SECOND LETTER A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER, INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. Litters were not cross-fostered; values excluded from group averages. C. vehicle control group dams cross-fostered with vehicle control litters. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 2): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS ------------------------------------------------ -------------------------------------------------------------- PATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 1 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP II a ----------------------------------------------- 1.6 MG/KG/DAY SUBSET C POSTPARTUM DAY --------------------------------------------------------------- 18943 18947 18948 18957 18960 18964 18967 18968 5.7 S.8 6.1 5.7 5.8 5.7 6.0 5.4 5.7 5.3 5.7 5.6 5.6 5.5 5.8 S.6 5.4 6.1 5.9 S.9 5@B 5.8 6.2 5.4 5.7 5.6 5.7 5.5 5.7 6.1 5.5 5.9 5.6 5.9 5.8 S.S 6.1 5.8 6.2 5.6 5.8 4.4 5.5 5.9 5.2 5 6.2 6.7 6.2 5.6 6.1 5.8 5.8 5.9 5.8 5.6 5.7 5.5 5.5 5.2 5.5 5.4 5.2 5.7 5.3 5.2 5.6 5.4 5.4 5.2 5.3 5.7 5.5 4.7 6.0 6.4 6.2 6.3 6.0 5.9 6.0 5.9 6.1 5.8 5.6 6.5 5.8 5.8 6.1 6.2 6.2 5.8 5.9 5.9 5.9 5.8 5.5 5.5 5.7 5.5 5.6 5.8 S.4 5.1 5.2 7.3 6.0 6.3 6.1 6.0 5.6 6.6 6.9 6.1 5.9 18969 5.9 5.6 5.8 5.6 6.0 5.7 5.S 5.3 5.5 5.6 5.8 5.6 5.7 5.6 5.4 5.6 5 18970 5.7 5.5 5.8 5.7 5.8 5.8 5.6 5.4 6.0 5.6 5.3 5.4 5.6 5.1 5.3 18973 5.8 5.9 5.3 S.2 6.1 5.4 5.8 5.4 S.6 5.7 5.4 5.5 5.2 5.6 5.5 18974 5.6 5.5 5.9 5.9 5.6 S.S S.4 5.2 5.3 5.4 5.3 4.6 5.4 5.3 5.6 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP Il b 1.6 MG/KG/DAY SUBSET D POSTPARTUM DAY --------------------------------------------------------------------------------------------------------------- 18946c 7.0 7.2 6.6 6.8 6.7 7.0 7.3 6.6 6.6 6.4 6.6 6.1 6.2 6.4 6.7 d 18950 6.5 6.7 7.0 7.0 7.9 7.3 7.3 7.1 6.3 7.0 6.4 6.6 6.0 6.4 5.7 6.2 6 18951 8.4 7.7 7.2 7.8 7.9 7.8 7.3 7.9 7.4 7.6 7.6 7.8 7.0 7.7 18952 5.6 6.3 6.6 6.4 6.S 6.8 6.3 6.4 5.7 6.1 6.1 6.1 5.7 5.9 6.2 5.8 5 189S3 6.9 7.1 6.7 7.3 7.2 6.9 6.7 7.3 6.9 6.1 7.1 6.8 6.4 6.1 6.3 6.6 18958 7.1 7.4 6.8 7.1 7.3 7.2 6.3 5.5 6.4 6.8 6.7 6.5 7.2 6.0 18959 6.1 6.2 6.0 6.0 6.4 6.0 5.9 5.8 6.0 5.5 5.3 5.9 5.8 5.5 6.1 4.1 18961 6.1 6.2 6.9 6.1 6.2 6.5 5.8 6.1 6.4 6.3 6.3 6.4 6.2 5.4 5.5 6.0 18963 6.1 5.6 5.6 5.7 5.9 6.0 5.8 5.5 6.0 5.7 5.6 5.2 5.8 5.5 5.6 5.5 5 18965 6.4 6.0 6.1 5.9 5.8 5.9 5.5 5.9 6.1 5.4 5.2 5.3 6.0 5.9 5.8 5.9 6 18972 6.4 6.1 6.2 6.4 6.4 6.7 6.3 5.7 6.2 S.9 6.1 5.7 5.4 5.6 5.9 5.6 18976 6.2 6.0 6.1 5.9 6.0 6.3 6.3 6.5 5.8 5-9 6.2 5.9 5.4 5.7 6.0 6.1 5 18977c 6.7 7.1 6.1 6.5 6.4 7.0 7.0 6.1 5.9 6.3 6.1 FD 1 --------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST LETTER M-MALE, F-FEMALE SECOND NUMBER LETTER FOLLOWING A-ALIVE, "SECOND S-STILLBORN, D-DIED, LETTER" INDICATES THE C-CULLED, M-MISSING DAY POSTPARTUM THE (PRESUMED CANNIBALIZED) EVENT OCCURRED. a. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group'litters. c. Litters were not cross-fostered; d. Value was not recorded. values excluded from group averages. PROTOCOL 418-014: ORAL IGAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 3): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 1 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP I a 0 (VEHICLE) MG/KG/DAY SUBSET A POSTPARTUM DAY --------------------------------------------------------------------------------------------------------------- 18903 4.7 5.9 5.7 6.5 6.5 6.0 MM 3 6.8 6.5 6.5 5.3 6.0 5.8 4.6 FM 3 18907 8.0 6.7 8.8 7.3 8.4 7.7 8.1 5.3 8.2 7.4 6.7 8.1 FD 4 18908 7.0 6.7 7.5 MM 4 7.4 7.6 5.7 7.2 6.8 7.6 6.5 6.3 7.0 8.0 7.3 6.5 18909 7.6 7.7 S.3 7.9 7.6 7.8 7.8 MD 4 8.0 8.1 7.0 7.5 4.6 7.0 7.3 7.8 18918b 18920 10.5 9.2 9.7 9.5 10.6 10.1 10.4 MM 2 MM 2 9.7 9.0 9.3 9.2 9.0 9.5 9.0 a 7.2 4.1 6.9 7.4 7.1 6.2 7.7 7.0 7.0 7.6 7.1 7.1 6.2 FM 3 FD 2 18924 8.2 8.4 7.3 7.5 7.7 6.5 7.0 MM 3 7.2 7.0 7.7 7.6 6.8 5.4 7.3 4.9 FD 18929 5.6 7.0 8.6 8.0 7.5 7.9 8.7 MM 3 7.3 6.0 6.8 7.5 6.4 8.2 7.4 7.9 18934 8.0 7.5 6.2 7.1 8.3 7.0 7.4 6.8 7.1 7.4 7.5 7.1 5.9 FD 2 18936b 8.4 9.2 9.2 8.9 8.4 8.2 8.6 7.2 8.1 7.7 7.8 FM 2 18937 8.1 8.2 8.3 7.7 7.7 7.4 7.1 7.7 7.4 7.1 7.7 6.4 7.8 18941 6.7 8.2 8.2 7.1 9.0 MM 3 MM 3 MD 2 9.3 6.1 8.7 3.6 FD 2 18942 5.7 7.6 7.9 6.5 7.2 7.3 7.1 6.8 6.7 6.9 6.9 6.1 7.2 6.4 7.9 6.9 7 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP I C 0 (VEHICLE) MG/KG/DAY SUBSET B POSTPARTUM DAY --------------------------------------------------------------------------------------------------------------- 18901 8.7 8.8 9.5 10.7 9.5 9.2 8.3 8.3 8.6 8.4 9.6 8.8 8.9 8.0 7.9 9.3 18904 6.8 6.5 9.2 7.8 7.9 7.7 7.2 6.3 6.8 8.6 7.5 7.7 6.4 6.8 6.3 18906 18912 8.0 8.3 8.8 6.7 8.4 7.7 9.0 8.4 7.7 6.0 8.8 6.7 7.9 7.6 7.8 8.4 8 6.5 7.2 5.5 7.0 6.5 7.4 6.9 7.3 7.4 7.4 6.1 6.8 5.9 6.8 6.1 6.7 6 18917 9.3 9.0 9.0 8.2 7.1 9.1 8.7 8.3 9.3 8.9 8.4 8.6 8.1 7.7 8.5 8.5 18919 10.2 9.6 8.7 9.4 9.0 10.1 9.3 9.5 9.1 9.5 9.3 9.1 18922 8.1 10.1 8.8 8.7 8.7 8.0 8.5 8.3 8.7 9.2 9.4 9.5 8.4 7.3 7.9 18923 9.9 9.9 8.4 9.3 10.6 9.5 9.4 8.1 9.3 8.6 9.4 9.7 9.2 7.6 9.2 FM 3 18931 8.3 7.7 8.6 8.2 8.3 8.6 8.2 8.5 7.5 7.9 8.3 6.2 8.0 7.2 7.6 8.7 8 18932 9.3 8.9 9.8 9.0 8.8 9.0 8.5 9.2 8.5 8.4 8.5 8.5 8.7 8.2 18935 9.0 8.7 7.8 8.8 8.7 8.0 7.5 8.7 8.2 8.2 8.0 4.6 8.2 8.7 7.5 7.3 18938 8.5 9.5 8.5 8.9 8.5 8.1 8.2 9.1 8.1 7.7 9.1 8.6 7.4 8.2 8.5 --------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST LETTER M-MALE, F-FEKALE SECOND LETTER A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. Vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. Litters were not cross-fostered; values excluded from group averages. c. vehicle control group dams cross-fostered with vehicle control litters. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 4): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS ---------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING ---------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 is 16 17 ---------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP Il a 1.6 MG/KG/DAY SUBSET C POSTPARTUM DAY ---------------------------------------------------------------------------------------------------------------- 18943 18947 7.1 7.8 8.7 7.5 8.8 8.2 8.6 8.8 7.3 8.2 8.2 8.5 7.6 6.9 8.4 7.6 8.0 7.7 7.4 7.0 7.9 7.9 7.4 6.3 7.6 7.7 7.0 7.1 6.7 16948 18957 6.1 7.1 6.1 7.1 4.4 7.2 MM 4 MM 3 MD 2 MD 2 MD 2 6.5 5.1 FD 3 FD 3 FM 3 FM 8.8 7.0 7.1 MM 3 6.9 6.8 6.9 7.3 8.7 7.7 7.6 7.7 FM 3 10960 6.6 7.3 6.5 MM 4 MM 4 6.0 6.4 7.4 6.5 6.5 6.6 4.6 5.2 FM 4 FM 4 18964 18967 18968 6.9 6.6 6.6 6.5 7.1 6.2 MM 3 6.3 6.0 7.4 6.3 6.1 7.0 7.6 6.6 9.0 8.6 7.8 6.7 8.3 9.0 7.7 6.6 8.1 7.5 7.3 7.7 7.9 7.6 6.1 6.0 10.4 MD 2 9.3 8.3 9.4 9.4 9.1 8.5 7.6 8.0 18969 18970 7.4 7.9 6.8 7.2 7.6 7.6 6.8 7.3 7.4 7.5 6.9 8.2 7.9 7.5 7.8 8.2 6. 7.6 7.1 7.1 7.8 6.2 6.9 7.7 6.1 7.7 6.9 4.3 3.9 6.0 6.7 FM 3 18973 6.1 8.7 7.7 7.8 7.3 6.7 7.0 7.8 7.8 6.8 6.2 7.5 7.2 7.8 FD 2 18974 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FM 4 FM 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 ---------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP II b 1.6 MG/KG/DAY SUBSET D POSTPARTUM DAY ---------------------------------------------------------------------------------------------------------------- 18946c 18950 18951 18952 18953 18958 18959 18961 18963 18965 18972 18976 8.7 6.0 4.3 MD 4 MD 4 MM 3 MD 2 MD 2 MD 2 5.2 FM 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 7.3 7.9 7.5 8.3 8.7 7.8 8.3 9.1 6.7 7.2 6.7 8.8 7.4 7.4 7.3 6.3 7. 8.8 8.9 8.0 8.6 8.9 9.6 8.8 8.6 8.9 8.3 8.1 7.5 8.7 8.6 8.7 6.3 8.9 8.5 8.3 7.8 8.9 8.4 7.1 6.6 7.8 7.5 7.6 7.7 7.7 7.4 7. 8.0 8.7 5.5 8.8 8.3 8.4 9.1 8.9 8.8 7.4 8.0 9.0 7.8 7.8 8.3 8.9 9.3 7.8 10.0 8.9 9.5 9.S 7.8 8.1 8.7 8.4 8.0 6.9 8.5 7.7 8.4 8.0 8.0 8.1 7.8 8.5 8.0 8.3 8.8 8.0 7.3 7.3 8.4 7.8 7.0 FM 3 7.8 8.9 8.4 8.0 8.5 6.0 9.1 7.1 6.8 8.5 7.6 7.S 8.1 8.3 6.3 7.7 7.1 7.0 6.8 7.4 7.0 7.3 6.5 6.7 8.0 7.1 7.3 6.5 5.6 6.4 6.9 6.8 6. 8.2 8.5 8.4 7.3 8.1 8.0 7.5 8.2 8.2 7.4 7.3 6.3 7.2 7.2 7.6 8.0 B. 8.6 8.5 9.7 9.1 8.7 9.1 9.1 8.3 8.5 7.4 8.1 8.2 7.4 7.2 8.6 9.0 8.0 8.0 6.8 7.0 7.7 7.3 7.3 MD 2 7.6 6.7 6.6 6.4 7.0 7.4 7.4 7.4 7. 18977C MD 3 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FD 2 FD 2 FD 2 FD 2 FD 1 ---------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST LETTER SECOND LETTER M-MALE, F-FEMALE A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. c. Litters were not cross-fostered; values excluded from group averages. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 5): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - PI GENERATION PUPS --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 is 16 1 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY SUBSET A POSTPARTUM DAY --------------------------------------------------------------------------------------------------------------- 18903 MC 4 5.9 5.7 6.5 6.5 6.0 mm 3 6.8 FC 4 6.5 FC 4 6.0 5.8 4.6 FM 3 18907 8.0 6.7 8.8 7.3 8.4 7.7 8.1 FC 4 8.2 7.4 6.7 FC 4 FD 4 18908 7.0 6.7 7.5 MM 4 7.4 7.6 FC 4 7.2 FC 4 FC 4 6.5 FC 4 FC 4 8.0 7.3 6.5 18909 7.6 7.7 MC 4 7.9 7.6 MC 4 7.8 MD 4 8.0 8.1 7.0 7.5 FC 4 FC 4 FC 4 7.8 18918b 18920 10.5 MC 4 9.7 9.5 MC 4 10.1 10.4 MM 2 MM 2 FC 4 9.0 9.3 FC 4 FC 4 9.5 9.0 a 7.2 MC 4 6.9 7.4 7.1 MC 4 7.7 MC 4 7.0 7.6 7.1 7.1 6.2 FM 3 FD 2 18924 MC 4 8.4 7.3 7.5 MC 4 MC 4 7.0 MM 3 7.2 7.0 7.7 7.6 6.8 FC 4 7.3 FC 4 FD 18929 MC 4 MC 4 8.6 8.0 7.5 7.9 8.7 MM 3 7.3 FC 4 FC 4 FC 4 6.4 8.2 7.4 7.9 18934 8.0 7.5 MC 4 7.1 8.3 7.0 MC 4 FC 4 7.1 7.4 7.5 7.1 5.9 FD 2 18936b 8.4 9.2 MC 4 8.9 8.4 8.2 8.6 7.2 8.1 7.7 7.8 FM 2 18937 8.1 8.2 8.3 7.7 7.7 7.4 FC 4 7.7 7.4 7.1 7.7 FC 4 PC 4 18941 8.7 8.2 8.2 7.1 9.0 MM 3 MM 3 MD 2 9.3 6.1 8.7 3.6 FD 2 18942 MC 4 7.6 7.9 MC 4 MC 4 MC 4 7.1 6.8 6.7 6.9 FC 4 FC 4 7.2 FC 4 7.9 6.9 FC --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY SUBSET B POSTPARTUM DAY --------------------------------------------------------------------------------------------------------------- 18901 MC 4 8.8 9.5 10.7 9.5 9.2 8.3 8.3 8.6 FC 4 FC 4 FC 4 FC 4 FC 4 7.9 9.3 18904 MC 4 MC 4 9.2 7.8 7.9 7.7 7.2 FC 4 FC 4 8.6 7.5 7.7 PC 4 6.8 6.3 18906 MC 4 6.3 8.6 MC 4 MC 4 7.7 9.0 8.4 FC 4 FC 4 8.8 FC 4 7.9 7.6 7.8 8.4 FC 18912 MC 4 7.2 MC 4 7.0 6.5 MC 4 6.9 MC 4 MC 4 7.4 MC 4 6.8 FC 4 6.8 FC 4 6.7 FC 18917 9.3 9.0 MC 4 MC 4 7.1 9.1 8.7 MC 4 MC 4 8.9 FC 4 8.6 8.1 7.7 FC 4 8.5 18919 MC 4 MC 4 8.7 9.4 9.0 10.1 9.3 9.5 9.1 9.5 9.3 9.1 18922 8.1 10.1 8.8 8.7 0.7 FC 4 FC 4 FC 4 8.7 9.2 9.4 9.5 FC 4 FC 4 7.9 18923 9.9 9.9 MC 4 9.3 1,0.6 9.5 FC 4 FC 4 9.3 8.6 FC 4 FC 4 9.2 7.6 9.2 FM 3 18931 8.3 7.7 8.6 MC 4 8.3 MC 4 8.2 8.5 FC 4 7.9 FC 4 FC 4 FC 4 FC 4 7.6 8.7 8 18932 9.3 MC 4 9.8 9.0 .0.8 9.0 8.5 9.2 8.5 FC 4 8.5 FC 4 8.7 FC 4 16935 9.0 8.7 MC 4 8.8 8.7 MC.4 7.5 8.7 FC 4 8.2 FC 4 FC 4 FC 4 8.7 7.5 7.3 18938 8.5 9.5 8.5 MC 4 MC 4 8.1 MC 4 9.1 8.1 MC 4 9.1 8.6 MC 4 8.2 8.5 --------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST LETTER M-MALE, F-FEMALE SECOND NUMBER LETTER FOLLOWING A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. vehicle control group dams cross-fostered 1.6 mg/kg/day dosage group litters. b. Litters were not cross-fostered; values excluded from group averages. c. vehicle control group dams cross-fostered vehicle control litters. PROTOCOL 418-014: OPAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 6): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS -------------- ------------------------------------------------------------------------------------------------ RATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 is 16 1 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP Il a 1.6 MG/KG/DAY SUBSET C POSTPARTUM DAY --------------------------------------------------------------------------------------------------------------- 18943 7.1 7.8 8.7 7.5 8.8 FC 4 FC 4 8.8 7.3 8.2 8.2 FC 4 FC 4 FC 4 8.4 18947 7.6 8.0 MC 4 MC 4 MC 4 7.9 7.9 7.4 FC 4 7.6 7.7 7.0 7.1 6.7 18948 6.1 7.1 6.1 7.1 4.4 7.2 MM 4 MM 3 MD 2 MD 2 MD 2 6.5 5.1 FD 3 FD 3 FM 3 FM 18957 8.8 7.0 7.1 MM 3 FC 4 6.8 6.9 7.3 8.7 7.7 7.6 7.7 FM 3 18960 6.6 7.3 6.5 MM 4 MM 4: 6.0 6.4 7.4 6.5 6.5 6.6 FC 4 5.2 PH 4 FM 4 18964 6.9 6.6 MC 4 MC 4 7.1 6.2 MM 3 6.3 FC 4 7.4 FC 4 6.1 7.0 7.6 6.6 18967 9.0 8.6 MC 4 MC 4 8.3 MC 4 7.7 MC 4 8.1 7.5 FC 4 FC 4 7.9 7.6 6.1 6.0 18968 10.4 MD 2 9.3 8.3 9.4 9.4 9.1 8.5 7.6 8.0 18969 MC 4 7.9 MC 4 MC 4 MC 4 7.6 MC 4 7.3 MC 4 7.5 MC 4 8.2 7.9 7.5 7.8 8.2 6 18970 MC 4 MC 4 7.1 7.8 MC 4F,; 6.9 7.7 MC 4 7.7 6.9 4.3 3.9 6.0 6.7 FM 3 18973 MC 4 8.7 MC 4 MC 4 7.3p, 6.7 MC 4 7.8 7.8 6.8 6.2 7.5 7.2 7.8 FD 2 18974 MD 2 MD 2 MD 2 MD 2 MD 2: MD 2 FM 4 FM 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP II b 1.6 MG/KG/DAY SUBSET D POSTPARTUM DAY --------------------------------------------------------------------------------------------------------------- 18946c 8.7 6.0 4.3 MD 4 MD 4; MM 3 MD 2 MD 2 MD 2 5.2 FM 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 18950 MC 4 MC 4 7.5 8.3 8.r-.-MC 4 8.3 9.1 FC 4 7.2 6.7 8.8 FC 4 7.4 PC 4 FC 4 7 18951 8.8 8.9 MC 4 MC 4 8.9 9.6 8.8 MC 4 8.9 8.3 FC 4 7.5 8.7 8.6 18952 8.7 MC 4 8.9 8.5 8.3 MC 4 8.9 MC 4 FC 4 FC 4 7.8 FC 4 7.6 7.7 7.7 7.4 FC 18953 8.0 8.7 MC 4 8.8 MC 4, 8.4 9.1 MC 4 8.8 FC 4 8.0 9.0 FC 4 7.8 FC 4 8.9 18958 9.3 7.8 10.0 8.9 MC 4 9.5 7.8 8.1 8.7 FC 4 8.8 PC 4 8.5 FC 4 18959 MC 4 8.0 8.0 8.1 7.8 MC 4 8.0 MC 4 MC 4 8.0 FC 4 7.3 8.4 7.8 7.0 FM 3 18961 7.8 8.9 8.4 8.0 8.5 FC 4 9.1 FC 4 FC 4 8.5 FC 4 PC 4 8.1 8.3 8.3 FC 4 18963 MC 4 7.0 6.8 7.4 MC 4 7.3 MC 4 MC 4 8.0 MC 4 MC 4 6.S FC 4 FC 4 6.9 6.0 6 10965 8.2 8.5 8.4 MC 4 8.1 8.0 7.5 8.2 FC 4 FC 4 FC 4 FC 4 7.2 FC 4 7.6 8.0 FC 18972 8.6 MC 4 9.7 MC 4 8.7 9.1 9.1 8.3 FC 4 7.4 8.1 FC 4 FC 4 FC 4 8.6 9.0 18976 8.0 8.0 6.8 MC 4 7.7 MC 4 7.3 MD 2 7.6 6.7 FC 4 FC 4 FC 4 FC 4 7.4 FC 4 7 18977c MD 3 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FD 2 FD 2 FD 2 FD 2 FD 1 --------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST LETTER M-MALE, F-FEMALE SECOND LETTER A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. 1.6 mg/kg/day dosage group dame cross-fostered with 1.6 mg/kg/day dosage group litters. b. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. C. Litters were not cross-fostered; values excluded from group averages. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 7): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS -------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING -------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 a 9 10 11 12 13 14 15 16 -------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP I a 0 (VEHICLE) MG/KG/DAY SUBSET A POSTPARTUM DAY -------------------------------------------------------------------------------------------------------------- 18903 18907 MC 4 14.9 10.9 11.2 11.4 13.9 10.5 13.8 9.5 11.4 10.0 13.4 MM 3 12.0 10.2 FC 4 FC 4 11.8 8.1 11.5 FC 4 14.3 10.2 FC 4 11.5 FD 4 8.2 FM 3 18908 18909 18918b 18920 18924 18929 18934 18936b 12.8 12.2 12.7 MM 4 10.9 10.6 FC 4 12.4 FC 4 FC 4 11.7 FC 4 FC 4 12.7 12.1 11.3 14.0 13.0 MC 4 13.0 13.2 MC 4 13.6 MD 4 14.5 13.7 12.9 14.2 FC 4 FC 4 FC 4 11.8 16.7 MC 4 17.1 15.6 MC 4 16.7 16.5 MM 2 MM 2 FC 4 15.3 1S.6 FC 4 FC 4 13.7 15.1 1 12.6 MC 4 11.5 12.3 12.5 MC 4 12.4 MC 4 12.0 11.4 10.4 11.9 13.1 FM 3 FD 2 MC 4 13.6 12.6 11.0 MC 4 MC 4 13.0 MM 3 12.3 12.3 11.6 12.0 11.5 PC 4 12.7 FC 4 F MC 4 MC 4 14.2 13.9 13.3 14.6 12.5 MM 3 10.8 FC 4 FC 4 FC 4 12.2 13.0 12.5 13.8 14.5 11.9 MC 4 13.2 13.9 11.5 MC 4 FC 4 13.6 10.4 12.5 13.5 12.1 FD 2 13.5 13.1 MC 4 13.0 12.9 12.6 13.3 12.7 12.5 11.2 11.8 FM 2 18937 12.2 12.4 12.2 11.0 11.7 11.6 FC 4 11.7 11.2 11.1 11.8 FC 4 FC 4 18941 10.3 12.S 12.7 14.6 14.1 MM 3 MM 3 MD 2 9.1 13.7 15.2 FM 5 FD 2 18942 MC 4 12.5 12.7 MC 4 MC 4 MC 4 12.2 11.5 13.0 12.1 FC 4 FC 4 12.2 FC 4 12.2 13.5 F -------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP I c 0 (VEHICLE) MG/KG/DAY SUBSET B POSTPARTUM DAY -------------------------------------------------------------------------------------------------------------- 18901 MC 4 15.0 14.7 15.8 17.4 15.3 13.6 13.2 14.2 FC 4 FC 4 FC 4 FC 4 FC 4 15.5 14.9 18904 MC 4 MC 4 16.0 14.1 14.2 13.5 13.3 FC 4 FC 4 11.4 11.6 14.2 FC 4 12.3 13.3 18906 MC 4 13.7 12.6 MC 4 MC 4 13.7 13.9 13.6 FC 4 FC 4 13.8 FC 4 12.9 12.6 13.9 13.8 F 18912 MC 4 11.1 MC 4 13.5 12.3 MC 4 12.8 MC 4 MC 4 12.8 MC 4 12.1 FC 4 12.8 FC 4 12.8 F 18917 14.1 12.3 MC 4 MC 4 14.1 13.9 14.6 MC 4 MC 4 14.0 FC 4 14.6 13.0 13.1 FC 4 13.3 18919 MC 4 MC 4 14.0 17.7 14.7 16.1 15.7 15.4 14.7 15.1 15.4 15.0 18922 15.0 15.8 15.0 14.5 17.1 FC 4 FC 4 FC 4 16.0 16.4 15.0 13.7 FC 4 FC 4 14.3 18923 15.8 15.7 MC 4 16.3 17.6 15.1 FC 4 FC 4 15.2 14.5 FC 4 FC 4 13.9 12.8 15.0 FM 3 18931 12.5 14.2 14.1 MC 4 14.5 MC 4 13.7 14.5 FC 4 14.2 FC 4 FC 4 FC 4 FC 4 13.6 12.4 1 18932 15.3 MC 4 14.5 14.8 14.0 13.9 13.5 13.9 14.0 FC 4 13.7 FC 4 14.4 FC 4 18935 13.5 12.2 MC 4 13@B 14.1 MC 4 14.1 14.0 FC 4 11.9 FC 4 PC 4 FC 4 12.4 13.2 13.3 18938 13.2 12.8 14.7 MC 4 MC 4 14.5 MC 4 14.3 13.5 MC 4 13.4 14.2 MC 4 13.4 14.2 -------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST LETTER M-MALE, F-FEMALE SECOND LETTER A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. Litters were not cross-fostered; values excluded from group averages. c. vehicle control group dams cross-fostered with vehicle control litters. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 8): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 1 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP II a 1.6 MG/KG/DAY SUBSET C POSTPARTUM DAY --------------------------------------------------------------------------------------------------------------- 18943 14.1 12.0 12.4 13.6 12.6 PC 4 FC 4 12.9 14.7 12.7 12.5 FC 4 FC 4 FC 4 12.4 18947 12.2 13.3 MC 4 MC 4 MC 4 12.5 11.9 12.7 FC 4 12.7 12.7 11.0 11.7 10.3 18948 6.0 9.6 9.9 11.7 11.7 11.5 MM 4 MM 3 MD 2 MD 2 MD 2 9.7 8.2 FD 3 FD 3 FM 3 FM 18957 14.1 11.5 11.3 MM 3 FC 4 12.7 11.7 14.0 12.2 11.5 11.6 12.7 FM 3 18960 9.9 9.5 11.5 MM 4 MM 4 8.6 8.6 9.4 9.7 10.4 7.3 FC 4 11.2 FM 4 FM 4 18964 18967 11.0 9.7 MC 4 MC 4 9.9 10.6 MM 3 10.4 PC 4 11.5 FC 4 10.5 11.4 10.3 11.3 14.8 14.4 MC 4 MC 4 13.4 MC 4 13.4 MC 4 14.1 11.8 FC 4 FC 4 12.4 9.9 12.8 9.5 18968 15.4 MD 2 13.1 14.2 14.2 12.2 13.9 12.0 13.1 14.6 18969 MC 4 12.2 MC 4 MC 4 MC 4 12.6 MC 4 12.4 MC 4 12.6 MC 4 13.5 12.3 10.6 11.5 13.3 11 18970 MC 4 MC 4 13.3 13.4 MC 4 12.5 11.4 MC 4 12.3 10.6 11.5 6.9 6.3 11.8 FM 3 18973 MC 4 13.1 MC 4 MC 4 10.8 12.0 MC 4 9.9 12.0 10.5 10.7 12.7 10.8 9.4 FD 2 18974 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FM 4 FM 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP Il b 1.6 MG/KG/DAY SUBSET D POSTPARTUM DAY --------------------------------------------------------------------------------------------------------------- 18946c 10.1 14.6 MD 5 MD 4 MD 4 MM 3 MD 2 MD 2 MD 2 7.8 FM 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 18950 MC 4 MC 4 13.7 11.8 12.9 MC 4 14.3 13.8 FC 4 12.4 14.2 11.7 FC 4 11.8 FC 4 FC 4 11 18951 12.8 12.9 MC 4 MC 4 12.8 11.9 12.7 MC 4 11.8 10.0 FC 4 12.2 10.4 11.7 18952 14.6 MC 4 15.3 14.5 14.7 MC 4 14.7 MC 4 FC 4 FC 4 13.6 FC 4 13.5 13.3 13.5 12.4 FC 18953 11.7 12.7 MC 4 13.1 MC 4 13.1 12.7 MC 4 12.2 FC 4 11.8 12.S PC 4 11.7 FC 4 ll.S 189SB 11.0 12.8 13.6 13.6 MC 4 13.5 11.3 12.4 12.5 FC 4 12.4 FC 4 12.9 FC 4 18959 MC 4 13.3 12.2 13.1 13.1 MC 4 13.2 MC 4 MC 4 13.1 FC 4 12.3 12.4 13.1 11.5 FM 3 18961 14.2 13.7 13.2 12.4 12.9 FC 4 12.9 FC 4 FC 4 13.2 FC 4 FC 4 13.9 12.7 13.0 PC 4 18963 MC 4 13.6 12.5 10.3 MC 4 12.4 MC 4 MC 4 12.1 MC 4 MC 4 11.7 FC 4 FC 4 11.8 12.5 11 1896S 13.0 12.7 12.S MC 4 13.0 12.2 12.0 12.2 FC 4 FC 4 FC 4 FC 4 11.9 FC 4 13.2 12.0 FC 18972 13.4 MC 4 13.7 MC 4 15.7 15.5 15.7 12.0 FC 4 15.6 15.1 FC 4 FC 4 FC 4 14.3 12.8 18976 12.9 13.5 12.2 MC 4 11.8 MC 4 13.4 MD 2 11.4 11.4 FC 4 FC 4 FC 4 FC 4 13.7 PC 4 13 18977C MD 3 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FD 2 FD 2 FD 2 FD 2 FD 1 --------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST LETTER M-MALE, F-FEMALE SECOND LETTER A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. 1.6 mg/kg/day dosage group dams crosb-fostered with 1.6 mg/kg/day dosage group litters. b. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. c. Litters were not cross-fostered; values excluded from group averages. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 9): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS --------------------------------------------------------------------------------------------------------------- PATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 1 --------------------------------------------------------------------------------------------------------------- PAT/ LITTER MATERNAL DOSAGE GROUP I a 0 (VEHICLE) MG/KG/DAY SUBSET A POSTPARTUM DAY --------------------------------------------------------------------------------------------------------------- 18903 MC 4 26.5 23.4 23.2 26.2 24.4 MM 3 24.8 FC 4 22.4 FC 4 24.9 20.3 23.8 FM 3 18907 26.4 29.3 30.2 26.4 27.2 27.3 30.5 FC 4 26.4 2S.8 25.7 FC 4 FD 4 18908 25.8 27.2 27.5 MM 4 23.7 26.6 FC 4 25.3 FC 4 FC 4 28.0 FC 4 FC 4 26.1 25.4 28.1 18909 27.8 28.2 MC 4 28.1 28.8 MC 4 27.8 MD 4 29.8 26.8 28.1 29.4 FC 4 PC 4 FC 4 25.1 18918b 33.2 MC 4 33.9 31.0 MC 4 33.7 33.4 MM 2 MM 2 FC 4 32.3 30.3 FC 4 FC 4 30.6 28.7 31 18920 26.1 MC 4 25.4 26.4 24.3 MC 4 22.7 MC 4 24.9 26.7 24.4 24.4 22.0 FM 3 FD 2 18924 MC 4 23.5 27.2 25.5 MC 4 MC 4 25.9 MM 3 27.4 26.0 25.6 2S.8 25.1 FC 4 23.9 FC 4 FD 18929 MC 4 MC 4 33.0 29.8 29.4 31.9 26.8 MM 3 28.6 FC 4 PC 4 FC 4 27.3 26.3 29.4 27.8 18934 28.7 24.4 MC 4 28.0 30.1 26.9 MC 4 FC 4 29.1 24.8 28.0 22.5 27.0 FD 2 18936b 25.1 2S.5 MC 4 24.8 25.8 24.5 24.8 21.7 22.5 24.0 23.7 FM 2 18937 27.0 25.5 24.5 25.2 22.5 24.5 FC 4 23.5 27.1 25.6 25.4 FC 4 FC 4 18941 30.8 31.3 28.2 27.1 23.7 MM 3 MM 3 MD 2 33.3 22.5 31.7 FM 5 FD 2 18942 MC 4 26.4 29.3 MC 4 MC 4 MC 4 26.6 29.1 28.5 26.7 FC 4 FC 4 28.1 FC 4 28.8 31.0 FC --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP I C 0 (VEHICLE) MG/KG/DAY SUBSET B POSTPARTUM DAY --------------------------------------------------------------------------------------------------------------- 18901 MC 4 3S.0 29.9 31.2 30.5 31.7 28.9 27.8 31.0 FC 4 FC 4 FC 4 FC 4 FC 4 31.6 31.1 18904 MC 4 MC 4 27.2 28.1 32.1 28.5 28.6 FC 4 FC 4 27.1 29.1 25.8 FC 4 24.9 25.7 18906 MC 4 27.9 29.0 MC 4 MC 4 30.4 28.7 27.3 FC 4 FC 4 27.5 FC 4 28.3 28.7 29.5 27.5 FC 18912 MC 4 25.8 MC 4 30.8 30.0 MC 4 30.3 MC 4 MC 4 31.9 MC 4 29.9 FC 4 29.9 FC 4 27.9 FC 18917 29.5 29.7 MC 4 MC 4 28.2 26.7 30.3 MC 4 MC 4 29.6 FC 4 30.3 26.6 28.3 PC 4 27.3 18919 MC 4 MC 4 28.6 31.3 30.2 33.3 31.3 30.6 30.6 30.6 28.9 28.8 18922 32.4 29.3 31.4 29.9 30.9 FC 4 FC 4 FC 4 29.1 32.6 32.6 29.1 FC 4 FC 4 31.8 18923 31.6 28.5 MC 4 30.1 31.0 31.2 FC 4 FC 4 25.7 28.7 FC 4 FC 4 30.5 28.7 26.8 FM 3 18931 31.5 30.9 29.9 MC 4 29.4 MC 4 31.5 32.3 FC 4 29.5 PC 4 FC 4 FC 4 FC 4 30.5 31.8 28 16932 27.1 MC 4 27.2 28.2 27.4 27.2 26.4 26.7 26.6 FC 4 27.7 FC 4 27.1 FC 4 18935 27.4 26.8 MC 4 27.7 26.6 MC 4 22.4 25.6 FC 4 26.4 FC 4 FC 4 FC 4 26.1 26.7 24.5 18938 28.2 30.1 29.5 MC 4 MC 4 27.9 MC 4 28.5 32.0 MC 4 29.1 27.9 MC 4 28.9 29.0 ---------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST LETTER M-MALE, F-FEt4ALE SECOND LETTER A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. Vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. Litters were not cross-fostered; values excluded from group averages. C. Vehicle control group dams cross-fostered with vehicle control litters. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 10): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS ---------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING ---------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 ---------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP II a 1.6 MG/KG/DAY SUBSET C POSTPARTUM DAY 1 ---------------------------------------------------------------------------------------------------------------- 18943 18947 29.8 25.5 26.8 25.5 28.7 MC 4 27.2 MC 4 29.3 MC 4 FC 4 28.0 PC 4 27.0 27.9 25.9 31.4 FC 4 27.7 23.4 26.6 25.9 FC 4 22.3 FC 4 25.5 FC 4 23.8 27.1 18948 18957 18960 18964 18967 18968 23.4 23.6 26.3 27.0 27.5 24.1 24.0 MM 3 26.6 14.2 FC 4@ 24.1 MM 4 25.1 MM 3 23.9 MD 2 26.3 MD 2 24.6 MD 2 25.8 22.5 27.7 19.1 FM 3 FD 3 FD 3 FM 3 FM 25.2 22.1 21.4 MM 4 MM 4 24.1 19.1 19.2 21.2 22.1 16.4 FC 4 21.1 FM 4 FM 4 21.8 21.0 MC 4 MC 4 20.1 23.1 MM 3 20.9 FC 4 21.9 PC 4 22.9 20.7 23.8 22.3 30.1 29.9 MC 4 MC 4 29.8 MC 4 28.8 MC 4 28.8 28.8 FC 4 FC 4 25.7 23.3 21.6 24.9 30.4 MD 2 26.0 25.6 24.6 27.9 30.1 27.6 27.0 24.4 18969 18970 18973 18974 MC 4 26.4 MC 4 MC 4 MC 4 26.5 MC 4 27.6 MC 4 26.0 MC 4 28.3 24.3 25.6 27.9 24.9 25. MC 4 MC 4 25.5 26.1 MC 4 24.8 22.8 MC 4 26.2 24.4 24.0 22.1 15.1 14.3 FM 3 MC 4 26.3 MC 4 MC 4 22.4 23.9 MC 4 20.0 23.8 21.5 24.3 23.4 21.3 22.7 FD 2 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FM 4 FM 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 ---------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP II b 1.6 MG/KG/DAY SUBSET D POSTPARTUM DAY 1 ---------------------------------------------------------------------------------------------------------------- 18946c 18950 18951 35.4 MC 4 24.6 25.2 MC 4 23.6 MD 5 24.0 MC 4 MD 4 28.4 MC 4 MD 4 28.6 24.1 MM 3 MC 4 24.6 MD 2 25.0 24.3 MD 2 28.4 MC 4 MD 2 FC 4 20.6 19.3 23.0 21.6 FM 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 24.5 2S.3 PC 4 25.9 FC 4 FC 4 28. FC 4 23.4 22.7 19.9 18952 18953 18958 32.4 MC 4 28.7 31.5 29.8 MC 4 31.0 MC 4 FC 4 FC 4 28.0 FC 4 27.5 28.0 28.9 28.3 FC 25.9 22.0 MC 4 25.8 MC 4 25.5 27.6 MC 4 24.5 FC 4 25.5 24.5 FC 4 25.3 FC 4 24.3 26.3 21.5 24.2 25.1 MC 4 26.6 24.9 24.5 22.0 FC 4 24.0 FC 4 24.5 FC 4 18959 18961 18963 18965 18972 18976 18977c MC 4 26.4 MC 4 26.4 29.2 27.2 MD 3 26.9 26.9 28.7 24.9 MC 4 25.8 MD 2 26.1 26.8 26.5 25.8 29.2 24.7 MD 2 27.6 25.6 27.5 MC 4 MC 4 MC 4 MD 2 27.5 25.6 MC 4 25.3 33.8 26.2 MD 2 MC 4 FC 4 24.8 25.5 33.7 MC 4 MD 2 27.3 26.1 MC 4 24.7 31.2 27.0 MD 2 MC 4 FC 4 MC 4 24.3 30.7 MD 2 FD 2 MC 4 FC 4 26.7 FC 4 FC 4 24.3 FD 2 26.7 23.7 MC 4 FC 4 28.1 23.4 FD 2 FC 4 FC 4 MC 4 FC 4 33.6 FC 4 FD 2 26.0 FC 4 27.1 FC 4 FC 4 FC 4 FD 1 24.9 25.7 FC 4 24.7 FC 4 FC 4 2S.4 27.3 FC 4 FC 4 FC 4 FC 4 28.0 25.6 24.8 26.1 30.6 25.1 FM 3 FC 4 25.7 26. 24.6 FC 26.4 FC 4 26. ---------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST LETTER M-MALE, F-FEMALE SECOND LETTER A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. c. Litters were not cross-fostered; values excluded from group averages. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 11): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS ---------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING ---------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 ---------------------------------------------------------------------------------------------------------------- PAT/ LITTER MATERNAL DOSAGE GROUP I a 0 (VEHICLE) MG/KG/DAY SUBSET A POSTPARTUM DAY 2 --------------------------------------------------------------------------------------------------------------- 18903 18907 MC 4 41.6 44.6 46.3 41.8 4S.8 40.2 43.3 39.6. 39.2 43.7 40.5 MM 3 42.1 37.3 FC 4 FC 4 46.1 38.7 42.1 FC 4 42.3 40.1 FC 4 39.5 FD 4 35.5 FM 3 18906 18909 18918b 18920 18924 40.9 42.9 42.4 MM 4 42.6 42.4 FC 4 36.6 FC 4 FC 4 39.3 FC 4 PC 4 37.3 39.9 41.3 43.2 43.9 MC 4 44.1 41.8 MC 4 40.6 MD 4 41.1 38.9 37.6 43.5 FC 4 PC 4 FC 4 41.2 53.9 MC 4 S7.2 53.2 MC 4 56.0 29.7 MM 2 MM 2 FC 4 49.3 51.5 FC 4 PC 4 47.5 51.2 50. 43.5 MC 4 43.2 41.4 37.9 MC 4 40.8 MC 4 40.6 42.9 41.2 40.6 37.3 FM 3 FD 2 MC 4 42.6 39.8 40.4 MC 4 MC 4 39.2 MM 3 44.0 40.9 41.3 41.0 41.7 PC 4 39.6 FC 4 FD 18929 MC 4 MC 4 49.2 47.3 42.7, 46.7 50.1 MM 3 41.2 FC 4 FC 4 FC 4 45.2 42.2 43.2 47.0 18934 43.5 41@5 MC 4 38.6 44.1 42.5 MC 4 FC 4 43.3 40.8 40.9 38.9 37.3 FD 2 18936b 30.7 39.8 MC 4 39.5 40.4 41.6 39.2 35.6 35.1 39.2 36.4 FM 2 18937 18941 42.5 41.7 37.5 40.3 40.1 45.4 39.9 47.6 41.7 47.3 43.3 MM 3 FC 4 MM 3 41.1 MD 2 40.2 48.8 39.3 48.0 41.1 39.4 FC 4 FM 5 PC 4 FD 2 18942 MC 4 43.0 42.9 MC 4 MC 4 MC 4 40.0 41.7 40.7 41.0 FC 4 FC 4 41.8 PC 4 44.5 39.2 FC ---------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY SUBSET B POSTPARTUM DAY 2 ---------------------------------------------------------------------------------------------------------------- 18901 MC 4 50.2 49.9 48.9 S4.6 46.6 44.2 47.S 42.5 FC 4 FC 4 FC 4 FC 4 FC 4 48.8 48.8 18904 MC 4 MC 4 41.1 44.5 45.2. 48.7 42.6 FC 4 FC 4 39.8 41.3 42.2 FC 4 43.9 40.2 18906 MC 4 49.6 53.0 MC 4 MC 4 50.6 48.3 48.2 FC 4 FC 4 48.8 FC 4 49.8 47.3 46.8 46.8 FC 18912 MC 4 48.7 MC 4 44.3 50.9. MC 4 48.4 MC 4 MC 4 50.9 MC 4 49.6 FC 4 47.1 FC 4 43.3 FC 18917 42.3 43.4 MC 4 MC 4 47.2 47.3 46.4 MC 4 MC 4 45.7 FC 4 48.5 45.8 43.9 FC 4 44.6 18919 MC 4 MC 4 47.2 45.4 51.8 51.1 49.5 49.4 50.0 49.1 46.7 43.9 18922 49.2 48.2 51.1 49.7 45.3 FC 4 FC 4 FC 4 50.6 45.3 50.8 45.0 FC 4 FC 4 50.2 18923 49.6 50.6 MC 4 50.3 50.2 51.4 FC 4 FC 4 44.7 49.3 FC 4 FC 4 44.9 46.2 45.5 FM 3 18931 50.5 50.2 45.9 MC 4 49.5, MC 4 48.2 50.9 FC 4 46.9 FC 4 PC 4 FC 4 FC 4 48.4 44.0 48. 18932 43.4 MC 4 43.8 44.3 44.4 42.8 44.0 43.7 42.2 FC 4 44.2 FC 4 43.2 FC 4 18935 44.2 42.2 MC 4 44.0 45.2 MC 4 36.1 41.1 FC 4 41.0 FC 4 FC 4 FC 4 42.9 39.9 42.8 18938 45.4 44.8 43.2 MC 4 MC 4- 43.5 MC 4 46.6 45.9 MC 4 43.0 43.3 MC 4 45.5 42.0 ---------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST LETTER M-MALE, F-FEMALE SECOND LETTER A-ALIVE, S-STILLBORN, D-DIED, NUMBER FOLLOWING "SECOND LETTER' INDICATES THE C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) DAY POSTPARTUM THE EVENT OCCURRED. a. Vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. Litters were not cross-fostered; values excluded from group averages. c. Vehicle control group dams cross-fostered with vehicle control litters. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 12): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS ---------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING ---------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 ---------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP 11 a 1.6 MG/KG/DAY SUBSET C POSTPARTUM DAY 2 ---------------------------------------------------------------------------------------------------------------- 18943 42.8 42.3 45.8 39.8 44.7 FC 4 FC 4 41.8 41.6 42.1 37.9 FC 4 FC 4 FC 4 46.4 18947 45.4 41.5 MC 4 MC 4 MC 4 40.8 45.0 42.7 FC 4 41.9 38.3 36.1 40.4 41.7 18948 40.7 36.5 37.9 40.7 39.5 25.8 MM 4 MM 3 MD 2 MD 2 MD 2 36.3 30.9 FD 3 FD 3 FM 3 FM 18957 42.5 39.1 37.4 MM 3 PC 4 41.4 40.1 38.0 38.8 37.7 43.9 39.0 FM 3 18960 34.1 39.5 34.1 MM 4 MM 4 33.4 35.6 36.9 27.0 33.2 30.4 PC 4 30.3 FM 4 FM 4 18964 33.1 32.0 MC 4 MC 4 35.2 35.0 MM 3 34.3 FC 4 35.1 FC 4 33.7 35.7 35.6 34.8 18967 45.8 46.5 MC 4 MC 4 47.9 MC 4 47.8 MC 4 47.7 40.7 FC 4 FC 4 39.3 39.2 45.1 41.1 18968 50.7 MD 2 38.0 44.1 40.1 45.7 40.5 43.5 43.1 40.2 18969 MC 4 40.9 MC 4 MC 4 MC 4 39.2 MC 4 42.4 MC 4 40.0 MC 4 40.2 37.6 37.2 40.2 38.0 38. 18970 MC 4 MC 4 39.8 41.4 MC 4 40.6 37.0 MC 4 40.7 37.0 27.0 37.5 36.1 27.2 FM 3 18973 MC 4 37.2 MC 4 MC 4 42.2 33.1 MC 4 34.1 37.4 35.3 38.0 37.8 37.4 33.2 FD 2 18974 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FM 4 FM 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 ---------------------------------------------------------------------------------------------------------------- PAT/ LITTER MATERNAL DOSAGE GROUP Il b 1.6 MG/KG/DAY SUBSET D POSTPARTUM DAY 2 ---------------------------------------------------------------------------------------------------------------- 18946c 55.0 42.S MD 5 MD 4 MD 4 MM 3 MD 2 MD 2 MD 2 36.3 FM 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 18950 MC 4 MC 4 47.3 42.8 39.9 MC 4 47.7 44.6 FC 4 40.3 37.7 37.0 FC 4 44.1 FC 4 FC 4 39. 18951 39.7 40.3 MC 4 MC 4 38.9 40.4 37.9 MC 4 34.5 38.5 FC 4 36.3 38.7 34.6 18952 49.8 MC 4 48.7 46.7 44.3 MC 4 46.1 MC 4 FC 4 FC 4 43.6 FC 4 43.1 44.3 41.6 44.4 FC 18953 44.8 37.3 MC 4 41.0 MC 4 44.0 40.9 MC 4 40.4 FC 4 42.9 38.6 FC 4 39.0 FC 4 40.7 18958 42.8 41.4 42.1 34.2 MC 4 40.6 41.5 39.0 39.9 FC 4 40.5 FC 4 36.4 FC 4 18959 MC 4 42.6 42.6 43.6 43.1 MC 4 42.3 MC 4 MC 4 39.8 FC 4 40.5 45.6 43.7 40.9 FM 3 18961 42.4 40.8 42.5 42.4 41.9 FC 4 41.0 FC 4 FC 4 42.6 FC 4 FC 4 39.8 36.8 40.5 FC 4 18963 MC 4 42.8 46.2 43.0 MC 4 41.6 MC 4 MC 4 42.4 MC 4 MC 4 40.7 FC 4 FC 4 42.8 41.7 41. 18965 45.5 44.1 43.0 MC 4 43.6 43.6 43.6 41.2 FC 4 FC 4 FC 4 FC 4 42.7 FC 4 43.0 41.0 FC 18972 43.6 MC 4 49.7 MC 4 42.0 46.8 48.5 43.4 FC 4 44.5 47.0 FC 4 FC 4 FC 4 42.7 40.7 18976 40.3 42.4 42.5 MC 4 40.7 MC 4 42.6 MD 2 39.0 42.3 FC 4 FC 4 FC 4 FC 4 39.9 FC 4 39. 18977c MD 3 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FD 2 FD 2 FD 2 FD 2 FD 1 ---------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST LETTER M-MALE, F-FEMALE SECOND LETTER A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. c. Litters were not crosa-fostered; values excluded from group averages. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 13): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS ----------------------------- I ---------------------------------------------------------------------------------- RATS ASSIGNED TO PHARAMACOKINETIC EVALUATION ---------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 ---------------------------------------------------------------------------------------------------------------- RAT/ LITTER 14ATERNAL DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY POSTPARTUM DAY I ---------------------------------------------------------------------------------------------------------------- 18910 7.3 7.4 7.6 7.2 7.0 18911 6.6 5.9 7.1 7.0 7.0 6.2 7.5 6.3 6.8 6.4 6.3 6.8 6.2 6.4 6.3 6.6 18913 6.2 6.8 6.8 6.5 5.3 6.9 6.4 6.2 6.1 5.9 5.9 5.6 6.3 5.9 7.0 6.0 6. 18915 6.3 6.3 6.8 6.0 5.9 6.1 6.1 5.5 6.5 5.5 6.2 6.5 6.0 6.3 6.4 FS FS 18926 7.2 7.2 7.4 7.4 6.2 6.7 7.2 6.7 6.7 7.2 7.4 6.5 18927 6.5 6.1 7.1 6.8 6.8 6.1 7.2 6.4 6.8 5.1 6.3 6.4 5.8 6.2 6.2 5.5 6. 18939 6.0 6.4 6.3 6.0 6.6 6.8 6.6 5.9 5.9 5.7 6.0 5.9 6.4 5.8 6.1 18940 6.9 6.2 6.5 6.3 6.2 6.2 6.5 6.1 5.5 5.8 6.0 5.8 5.6 6.1 6.2 5.4 ---------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP 11 1.6 MG/KG/DAY POSTPARTUM DAY 1 ---------------------------------------------------------------------------------------------------------------- 18956 6.7 6.4 6.2 6.3 6.6 ms 6.1 6.0 6.9 6.3 6.7 4.3 6.1 18971 ms 7.6 FS ---------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST SECOND LETTER LETTER M-MALE, F-FEMALE A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. PROTOCOL 418-014: OPAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 14): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM INDIVIDUAL DATA - Fl GENERATION PUPS ------------- - - ----------------------- RATS ASSIGNED TO PHARAMACOKINETIC EVALUATION -------------------------------------------- ---------------------------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 is 16 1 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY POSTPARTUM DAY 4 --------------------------------------------------------------------------------------------------------------- 18910 NO PUP WEIGHTS RECORDED 18911 18913 18915 8.4 9.5 9.5 9.1 10.3 7.9 7.4 7.9 9.5 9.3 9.5 8.8 8.6 8.7 8.8 7.2 9.9 7.4 8.7 7.6 7.9 6.5 6.7 7.2 8.2 8.6 8.2 7.2 8.6 8.8 8.8 MD 4 7 NO PUP WEIGHTS RECORDED 18926 18927 18939 10.6 11.6 15.7 13.0 11.4 9.3 11.9 12.9 13.9 11.4 11.5 12.6 8.4 8.8 8.6 9.1 8.8 9.3 8.3 9.2 8.7 7.5 8.2 7.0 9.0 8.0 7.6 9.3 FM NO PUP WEIGHTS RECORDED 18940 9.5 8.5 9.2 7.1 8.5 8.2 8.5 8.2 9.1 7.8 7.4 8.9 8.7 8.5 7.0 9.3 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP 11 1.6 MG/KG/DAY POSTPARTUM DAY 4 --------------------------------------------------------------------------------------------------------------- 18956 18971 8.8 9.3 9.2 9.6 9.6 ms ms 9.1 FS 9.1 8.9 9.2 6.7 8.9 9.6 9.4 --------------------------------------------------------------------------------------------------------------ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST SECOND NUMBER LETTER LETTER FOLLOWING M-KALE, F-FEMALE A-ALIVE, S-STILLBORN, D-DIED, "SECOND LETTER" INDICATES THE C-CULLED, M-MISSING DAY POSTPARTUM THE (PRESUMED CANNIBALIZED) EVENT OCCURRED. 7 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 15): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO PHARAMACOKINETIC EVALUATION --------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 1 --------------------------------------------------------------------------------------------------------------- PAT/ LITTER MATERNAL DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY POSTPARTUM DAY 7 ---------------------------------------------------------------------------------------------------------------- 18910 18.8 16.5 17.1 16.0 16.9 18911 18913 18915 18926 13.2 14.1 13.6 16.4 12.0 12.6 12.4 15.9 13.7 9.7 11.3 16.1 9.9 10.0 13.3 16.6 11.2 9.3 12.4 15.4 9.9 11.1 13.5 14.9 11.7 9.0 10.6 15.4 13.7 11.7 9.2 14.8 12.7 12.1 14.2 15.9 13.8 11.5 14.9 16.2 14.5 13.2 13.1 15.1 13.0 12.7 12.0 12.3 FM 7 9.7 10.4 7.3 MD 6 MD 4 10. 14.1 13.4 14.5 12.6 FS Fl 16.5 18927 12.3 12.0 12.2 12.7 11.7 12.9 11.9 12.4 10.7 10.8 12.4 9.4 10.2 11.0 12.4 9.1 FM 18939 12.2 13.5 14.1 12.9 13.2 12.7 12.0 12.4 11@9 12.9 13.3 12.2 13.1 13.3 11.1 18940 10.3 12.2 11.8 11.1 8.5 12.1 10.8 10.0 10.3 11.0 12.0 10.3 11.6 12.0 10.8 7.9 ---------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP Il 1.6 MG/KG/DAY POSTPARTUM DAY 7 ---------------------------------------------------------------------------------------------------------------- 18956 18971 13.4 13.2 13.0 13.2 12.7 ms ms 9.2 FS 12.9 12.4 12.4 13.3 12.1 11.8 9.7 ---------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST LETTER SECOND LETTER M-MALE, F-FEMALE A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 16): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO PHARAMACOKINETIC EVALUATION --------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 1 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY POSTPARTUM DAY 14 --------------------------------------------------------------------------------------------------------------- 18910 35.7 32.7 32.9 33.7 30.5 18911 18913 18915 18926 26.7 19.8 24.6 29.7 25.3 24.2 23.5 31.3 24.0 21.9 25.2 31.3 22.1 20.5 24.6 29.4 17.8 20.2 25.9 28.8 23.0 19.9 24.8 29.5 26.8 23.3 20.5 29.9 19.2 23.0 27.8 29.2 24.2 25.1 26.9 27.8 25.3 20.8 20.7 32.0 26.4 24.1 27.8 30.0 22.7 20.4 25.2 27.8 25.0 18.7 25.3 25.2 14.9 27.8 23.4 FM 7 ND 6 MD 4 21 25.2 FS p 18927 18939 22.6 23.2 23.8 21.7 22.5 20.8 23.7 21.3 21.2 22.6 20.6 19.S 22.7 18.0 20.0 FMIO FM 24.9 23.1 23.7 21.6 26.0 23.2 23.1 22.1 25.0 24.1 23.0 21.4 24.8 21.5 18.8 18940 18.3 16.0 22.3 24.S 16.6 20.2 17.4 18.9 23.7 20.3 13.1 20.6 20.0 19.2 19.0 20.9 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP 11 1.6 MG/KG/DAY POSTPARTUM DAY 14 --------------------------------------------------------------------------------------------------------------- 18956 18971 24.1 23.7 24.4 24.1 24.9 ms ms 11.7 FS 23.7 22.9 24.9 23.7 24.3 23.6 20.7 --------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST LETTER M-MALE, F-FEMALE SECOND LETTER A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 31 (PAGE 1): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATI -------------------------- -------------------- --- ------------------------- -------------------------------- RATS ASSIGNED TO CROSS-FOSTERING ------------------------------------------------- -- --------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 is 19 20 -------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP I a 0 (VEHICLE) MG/KG/DAY SUBSET A -------------------------------------------------------------------------------------------------------------- 18903 MC 4 M A M A M A M A M A MM 3 F A FC 18907 M A M A M A M A M A F A F A FC 4 F 18908 M A M A M A MM 4 F A F A FC 4 F A FC 18909 M A M A MC 4 M A M A MC 4 M A MD 4 F 18918b M A MC 4 M A M A MC 4 M A M A MM 2 MM 18920 M A MC 4 M A M A M A MC 4 M A MC 4 F 18924 MC 4 M A M A M A MC 4 MC 4 M A MM 3 M 18929 MC 4 MC 4 M A M A M A M A M A MM 3 F 18934 M A M A MC 4 M A M A M A MC 4 FC 4 F 18936b M A M A MC 4 M A M A M A M A F A F 18937 M A M A M A M A M A F A FC 4 F A F 18941 M A M A M A M A M A MM 3 MM 3 MD 2 F 18942 MC 4 M A M A MC 4 MC 4 MC 4 M A M A M --------------------------------------------------- 4 F A FC 4 F A F A F A FM 3 A F A F A FC 4 FD 4 4 FC 4 F A FC 4 FC 4 F A F A F A A F A F A F A FC 4 FC 4 FC 4 F A 2 FC 4 F A F A FC 4 FC 4 F A F A F A A P A F A F A F A FM 3 FD 2 A F A F A F A F A FC 4 F A FC 4 FD 4 A FC 4 FC 4 FC 4 F A F A F A F A A F A P A F A F A FD 2 A F A F A FM 2 A F A F A FC 4 FC 4 A F A F A FM 5 FD 2 A F A FC 4 FC 4 F A FC 4 F A F A FC 4 F A --------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP I C 0 (VEHICLE) MG/KG/DAY SUBSET 8 -------------------------------------------------------------------------------------------------------------- 18901 MC 4 M A M A M A M A M A F A F A F A FC 4 FC 4 FC 4 FC 4 FC 4 F A F A 18904 MC 4 MC 4 M A M A M A M A M A FC 4 FC 4 F A F A F A FC 4 F A F A 18906 MC 4 M A M A MC 4 MC 4 M A M A M A FC 4 FC 4 F A FC 4 F A F A F A F A FC 4 PC 4 FD 4 18912 MC 4 M A MC 4 M A M A MC 4 M A MC 4 MC 4 M A MC 4 F A FC 4 F A FC 4 F A FC 4 F A F A FM 4 18917 M A M A MC 4 MC 4 M A M A M A MC 4 MC 4 F A FC 4 F A F A F A FC 4 F A 18919 MC 4 MC 4 M A M A M A M A M A F A F A F A F A F A 18922 M A M A M A M A M A FC 4 FC 4 FC 4 F A F A F A P A FC 4 FC 4 F A 18923 M A M A MC 4 M A M A M A FC 4 FC 4 F A F A FC 4 FC 4 F A F A F A FM 3 18931 M A M A M A MC 4 M A MC 4 M A F A FC 4 F A FC 4 FC 4 FC 4 FC 4 F A F A F A 18932 M A MC 4 M A M A M A M A F A F A F A FC 4 F A FC 4 F A FC 4 1893S M A M A MC 4 M A M A MC 4 M A F A FC 4 F A PC 4 FC 4 FC 4 F A F A F A 18938 M A M A M A MC 4 MC 4 M A MC 4 M A M A MC 4 M A M A MC 4 F A F A -------------------------------------------------------------------------------------------------------------- FIRST LETTER M-MALE, F-FEMALE SECOND LETTER A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. Vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. Litters were not cross-fostered; values excluded from group averages. c. Vehicle control group dams cross-foatered with vehicle control litters. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 31 (PAGE 2): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATIO --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP II a 1.6 MG/KG/DAY SUBSET C --------------------------------------------------------------------------------------------------------------- 18943 M A M A M A M A M A FC 4 FC 4 F A F A F A F A PC 4 FC 4 FC 4 F A 18947 M A M A MC 4 MC 4 MC 4 M A M A M A FC 4 F A F A F A F A F A 18948 M A M A M A M A M A M A MM 4 MM 3 MD 2 MD 2 MD 2 F A F A FD 3 FD 3 FM 3 FM 3 18957 M A M A M A MM 3 FC 4 F A F A F A F A F A F A F A FM 3 18960 M A M A M A MM 4 MM 4 F A F A F A F A F A F A FC 4 F A FM 4 FM 4 18964 M A M A MC 4 MC 4 M A M A MM 3 M A FC 4 F A FC 4 F A F A F A F A 18967 M A M A MC 4 MC 4 M A MC 4 M A MC 4 M A F A FC 4 FC 4 F A F A F A F A 18968 M A MD 2 F A F A F A F A F A F A F A F A 18969 MC 4 M A MC 4 MC 4 MC 4 M A MC 4 M A MC 4 M A MC 4 M A F A F A F A F A F A 18970 MC 4 MC 4 M A M A MC 4 M A M A MC 4 M A F A P A F A F A F A FM 3 18973 MC 4 M A MC 4 MC 4 M A M A MC 4 M A M A M A F A F A F A F A FD 2 18974 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FM 4 FM 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER DOSAGE GROUP II b 1.6 MG/KG/DAY SUBSET D --------------------------------------------------------------------------------------------------------------- 18946c M A M A MD 5 MD 4 MD 4 MM 3 MD 2 MD 2 MD 2 F A FM 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 18950 MC 4 MC 4 M A M A M A MC 4 M A M A FC 4 F A F A F A FC 4 F A FC 4 FC 4 P A FC 4 18951 M A M A MC 4 MC 4 M A M A M A MC 4 F A F A FC 4 F A F A F A 18952 M A MC 4 M A M A M A MC 4 M A MC 4 FC 4 FC 4 F A FC 4 F A F A F A F A FC 4 189S3 M A M A MC 4 M A MC 4 M A M A MC 4 F A FC 4 F A F A FC 4 F A FC 4 F A 18958 M A M A M A M A MC 4 M A F A F A P A FC 4 F A FC 4 F A FC 4 189S9 MC 4 M A M A M A M A MC 4 M A MC 4 MC 4 F A FC 4 F A F A F A F A FM 3 18961 M A M A M A M A M A FC 4 F A FC 4 FC 4 F A FC 4 FC 4 F A F A F A FC 4 18963 MC 4 M A M A M A MC 4 M A MC 4 MC 4 M A MC 4 MC 4 F A FC 4 FC 4 F A F A F A F A FC 4 18965 M A M A M A MC 4 M A M A F A F A FC 4 FC 4 FC 4 FC 4 F A FC 4 F A F A FC 4 18972 M A MC 4 M A MC 4 M A M A M A F A FC 4 F A F A FC 4 FC 4 FC 4 F A F A 18976 M A M A M A MC 4 M A MC 4 M A MD 2 F A F A FC 4 FC 4 FC 4 FC 4 F A FC 4 F A F A 18977c MD 3 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FD 2 FD 2 FD 2 FD 2 FD I --------------------------------------------------------------------------------------------------------------- FIRST LETTER M-MALE, F-FEMALE SECOND LETTER A-ALIVE, S-STILLSORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. C. Litters were not cross-fostered; values excluded from group averages. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 31 (PAGE 3): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATIC ---------------------------------- -------------------------------------------------------------------------- RATS ASSIGNED TO PHARMACOKINETIC EVALUATION --------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 18910 M A MA M A FA FA 18911 M A M A M A M A M A M A M A M A M A M A M A F A F A F A F A FM 7 18913 M A M A M A M A M A M A M A M A M A M A M A M A M A M A MD 6 MD 4 F A F A 18915 M A M A M A M A F A F A F A F A F A F A FA F A F A F A FA FS FS 18926 M A M A M A M A M A M A F A F A F A FA F A F A 18927 M A M A M A M A M A M A M A M A F A F A F A F A P A F A F A FM10 FM 4 18939 M A M A M A M A M A M A M A F A P A F A F A F A FA F A FA 18940 M A M A M A M A M A M A M A M A M A M A F A F A F A F A FA FA --------------------------------------------------------------------------------------------------------------- RAT/ LITTER 14ATERNAL DOSAGE GROUP 11 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 18956 M A M A M A M A M A M S F A F A F A FA F A F A F A 18971 M S PA F S --------------------------------------------------------------------------------------------------------------- FIRST SECOND NUMBER LETTER LETTER FOLLOWING M-MALE, F-FEMALE A-ALIVE, S-STILLBORN, D-DIED, "SECOND LETTER" INDICATES THE C-CULLED, M-MISSING DAY POSTPARTUM THE (PRESUMED CANNIBALIZED) EVENT OCCURRED. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 32 (PAGE 1): CLINICAL OBSERVATIONS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PU ------------------------------------------- -------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING ---------------------------------------------------------------------------------------------------------------- SUBSET MATERNAL DOSAGE GROUP FOSTER MATERNAL DOSAGE (MG/KG/DAY) LITTER DAY(S) POSTPARTUM OBSERVATIONS a ---------------------------------------------------------------------------------------------------------------- A Ib 0 (VEHICLE) 18920 3 1/14 PUPS: NOT NURSING. B Ic 0 (VEHICLE) c II e 1.6 18941 18932 18968 18974 4 7-10 11-21 I- 2 3-21 2 1/9 PUPS: NOT NURSING. 1/10 PUPS: 1/10 PUPS: TIP OF TAIL, BLACK. TIP OF TAIL, BLACK. TAIL, BENT 2.5 CM FROM BASE OF TAIL).D 1/10 PUPS: 1/9 PUPS: RIGHT HINDPAW, RIGHT HINDPAW, SECOND DIGIT MIS SECOND DIGIT MISS 2/2 PUPS: NOT NURSING. D ii f 1.6 18950 7-15 2/10 PUPS: TIP OF TAIL, BLACK. 18953 1 1/16 PUPS: LABORED BREATHING. DARK IN COL ---------------------------------------------------------------------------------------------------------------- a. Tabulation restricted to adverse observations; all other pups appeared normal. b. vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. c. Vehicle control group dams cross-fostered with vehicle control group litters. d. observation confirmed at necropsy. e. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. f. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. PROTOCOL 418-014: OPAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 32 (PAGE 2): CLINICAL OBSERVATIONS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION -------------------------------------------------------------------------------------------------------------- PATS ASSIGNED TO PHARMACOKINETIC EVALUATION -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP LITTER MATERNAL DOSAGE (MG/KG/DAY) NUMBER DAY(S) POSTPARTUM OBSERVATIONS a -------------------------------------------------------------------------------------------------------------- I 0 (VEHICLE) 18911 14 1/15 PUPS: MOUTH, MASS (0.2 CM X 0.2 C14 18913 6 1/17 PUPS: NOT NURSING. -------------------------------------------------------------------------------------------------------------- a. Tabulation restricted to adverse observations; all other pups appeared normal. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 33 (PAGE 1): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fl GENERATION PUPS -------------------------------------------------------------------------------------------------------------- PUPS DYING PRIOR TO CROSS FOSTERING -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP LITTER MATERNAL DOSAGE (MG/KG/DAY) NUMBER DAY(S) POSTPARTUM OBSERVATIONS a -------------------------------------------------------------------------------------------------------------- I 0 (VEHICLE) 18904 1 3 PUPS: STILLBORN. 18912 1 18923 1 1 PUP: 1 PUP: STILLBORN. ALL TISSUES FOUND DEAD. 18937 1 1 PUP: STILLBORN. ALL TISSUES 1.6 18952 1 1 PUP: STILLBORN. ALL TISSUES 18973 1 1 PUP: FOUND DEAD. NO MILK IN BACK: RAISED AREA (2.0 0.3 CM), CONTAINED GAS. -------------------------------------------------------------------------------------------------------------- a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation individual pup clinical observations table (Table 32) for external observations confirmed at necropsy. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 33 (PAGE 2): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fl GENERATION PUPS ------------------------------------------------- ------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- SUBSET MATERNAL DOSAGE GROUP FOSTER MATERNAL DOSAGE (MG/KG/DAY) LITTER DAY(S) POSTPARTUM OBSERVATIONS a --------------------------------------------------------------------------------------------------------------- A Ib 0 (VEHICLE) 18903 4,21 ALL PUPS APPEARED NORMAL. 18907 4 21 1 PUP: FOUND DEAD. NO MILK IN S TISSUES APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 18908 4 1 PUP: FOUND DEAD. AUTOLYSIS PR EVALUATION. 18909 4 18918 21 ALL PUPS APPEARED ALL PUPS APPEARED NORMAL. NORMAL. 18920 2 4 1 PUP: FOUND DEAD. NO MILK IN S TISSUES APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 18924 4 21 1 PUP: FOUND DEAD. NO MILK IN S TISSUES APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 18934 2 21 1 PUP: FOUND DEAD. NO MILK IN S TISSUES APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 18936 21 ALL PUPS APPEARED NORMAL. 18941 2 2 PUPS: FOUND DEAD. ALL TISSUES 18942 4 ALL PUPS APPEARED NORMAL. --------------------------------------------------------------------------------------------------------------- a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. individual pup clinical observations table (Table 32) for external observations confirmed at necropsy. b. Vehicle control group dams cross-fostered 1.6 mg/kg/day dosage group litters. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 33 (PAGE 3): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fl GENERATION PUPS --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- SUBSET MATERNAL DOSAGE GROUP FOSTER MATERNAL DOSAGE (MG/KG/DAY) LITTER DAY(S) POSTPARTUM OBSERVATIONS a --------------------------------------------------------------------------------------------------------------- B Ib 0 (VEHICLE) 18901 18904 4,21 4 ALL PUPS APPEARED ALL PUPS APPEARED NORMAL. NORMAL. 18906 4 1 PUP: FOUND DEAD. AUTOLYSIS PR EVALUATION. 18917 21 ALL PUPS APPEARED NORMAL. 18922 21 ALL PUPS APPEARED NORMAL. 18923 21 ALL PUPS APPEARED NORMAL. 18931 21 ALL PUPS APPEARED NORMAL. 18932 4 ALL PUPS APPEARED NORMAL. 18935 21 ALL PUPS APPEARED NORMAL. --------------------------------------------------------------------------------------------------------------- a. complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. individual pup clinical observations table (Table 32) for external observations confirmed at necropsy. b. vehicle control group dams cross-fobtered vehicle control group litters. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 33 (PAGE 4): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fl GENERATION PUPS ---------------------------------- ----- -------------I ------------------------- ---------------------------- RATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- SUBSET MATERNAL DOSAGE GROUP FOSTER MATERNAL DOSAGE (MG/KG/DAY) LITTER DAY(S) POSTPARTUM OBSERVATIONS a --------------------------------------------------------------------------------------------------------------- c Il b 1.6 18943 21 10947 4 ALL PUPS APPEARED ALL PUPS APPEARED NORMAL. NORMAL. 18948 2 3 3 PUPS: 2 PUPS: FOUND DEAD. NO MILK IN TISSUES APPEARED NORMAL. FOUND DEAD. AUTOLYSIS P EVALUATION. 19857 4,21 ALL PUPS APPEARED NORMAL. 19860 19864 21 4,21 ALL PUPS APPEARED ALL PUPS APPEARED NORMAL. NORMAL. 19868 2 1 PUP: FOUND DEAD. NO MILK IN S TISSUES APPEARED NORMAL. 19870 21 ALL PUPS APPEARED NORMAL. 18973 2 4,21 1 PUP: FOUND DEAD. NO MILK ALL PUPS APPEARED NORMAL. IN S 18974 2 13 PUPS: FOUND DEAD. NO MILK IN OTHER TISSUES APPEARED --------------------------------------------------------------------------------------------------------------- a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. individual pup clinical observations table (Table 32) for external observations confirmed at necropsy. b. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. PROTOCOL 418-014; ORAL (GAVAGE) CROSS-FOSTERING SI'UDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 33 (PAGE 5): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fl GENERATION PUPS -------------------------------------------------- ------------------------------------------------------------ RATS ASSIGNED TO CROSS-FOSTERING --------------------------------------------------------------------------------------------------------------- SUBSET MATERNAL DOSAGE GROUP FOSTER MATERNAL DOSAGE (MG/KG/DAY) LITTER DAY(S) POSTPARTUM OBSERVATIONS a --------------------------------------------------------------------------------------------------------------- D II b 1.6 18946 2 2 PUPS: FOUND DEAD. ALL TISSUES 7 PUPS: FOUND DEAD. NO MILK IN TISSUES APPEARED NORMAL. 5 1 PUP: FOUND DEAD. ALL TISSUES 18951 18952 18953 18958 18961 18963 18965 18972 21 21 4,21 4,21 4,21 4 4 21 ALL PUPS APPEARED ALL PUPS APPEARED ALL PUPS APPEARED ALL PUPS APPEARED ALL PUPS APPEARED ALL PUPS APPEARED ALL PUPS APPEARED ALL PUPS APPEARED NORMAL. NORMAL. NORMAL. NORMAL. NORMAL. NORMAL. NORMAL. NORMAL. 18976 2 1 PUP: FOUND DEAD. NO MILK IN S TISSUES APPEARED NORMAL. 18977 2 11 PUPS: FOUND DEAD. NO MILK IN OTHER TISSUES APPEARED 3 1 PUP: FOUND DEAI). NO MILK IN S TISSUES APPEARED NORMAL. --------------------------------------------------------------------------------------------------------------- a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. individual pup clinical observations table (Table 32) for external observations confirmed at necropsy. b. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR-S STUDY NUMBER: T-6295.13) TABLE 33 (PAGE 6): NECROPSY OBSERVATIONS INDIVIDUAL DATA - F] GENERATION PUPS ----------------- ----------------- -- -------------1-@.----@ --------------------------- - --------- RATS ASSIGNED TO PHARMACOKINETIC EVALUATION --------------- ---------------------- MATERNAL DOSAGE GROUP LITTER ----------------- - - ------------------------------------------- MATERNAL DOSAGE (MG/KG/DAY) NUMBER DAY(S) POSTPARTUM OBSERVATIONS a --------------------------------------------------------------------------------------------------------------- I 0 (VEHICLE) 18910 14 18911 14 ALL PUPS APPEARED ALL PUPS APPEARED NORMAL. NORMAL. 18913 4 14 1 PUP: FOUND DEAD. AUTOLYSIS PR EVALUATION. ALL PUPS APPEARED NORMAL. 18915 1 14 2 PUPS: ALL PUPS STILLBORN. APPEARED NORMAL. 18926 14 18927 14 18939 14 18940 14 ALL PUPS APPEARED ALL PUPS APPEARED ALL PUPS APPEARED ALL PUPS APPEARED NORMAL. NORMAL. NORMAL. NORMAL. 1.6 18956 14 ALL PUPS APPEARED NORMAL. 18971 14 ALL PUPS APPEARED NORMAL. ---------------------------------------------------------------------------------------------------------------- a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. individual pup clinical observations table (Table 32) for external observations confirmed at necropsy. APPENDIX C PROTOCOL AND AMENDMENT 418-014:PAGE C-1 'i@-JPPJMMIC,A Argus Research Laboratories,Inc. 905 Sheehy Drive,BuildingA Horsham, PA 19044 Telephone: (215)443-8710 Telefax:(215)443-8587 PROTOCOL 418-014 SPONSOR'S STUDY NUMBER: T-6295.13 STUDY TITLE: Oral(Gavage)Cross-FostednSgtudyofPFOS inRats PURPOSE: The purposeofthistudyistoevaluatpeupsurvivaolfFl ratsfollowingPFOS treatmentof CD:CDOBR VAF/PlusO female ratsduringpremating,gestationand lactationF.l pups willbe cross-fosteredduringthe lactatiopneriodto differentiavtiea matemal milkthe dffferencefsorinutero exposure and exposure throughmatemal milk.Selected pharmacokineticsamples willbe collectefdrom Fo generationfemalesand Fl generationpups. TESTING FACILITY: Argus Research LaboratoriesI,nc. 905 Sheehy Drive,BuildingA Horsham, Pennsylvania 19044-1297 Telephone: (215)443-8710 Telefax: (215)443-8587 STUDY DIRECTOR: Raymond G. York,Ph.D.,DABT AssociateDirectorofResearch SPONSOR: 3M CorporateToxicology 3M Center,Building220-2E-02 St.Paul,Minnesota 55144-1000 STUDY MONITOR: MarvinT. Case, D.V.M.,Ph.D. Telephone: (651)733-5180 Telefax: (651)733-1773 ALTERNATE STUDY MONITOR: Andrew M. Seacat,Ph.D. Telephone: (651)575-3161 Telefax: (651)733-1773 418-014:PAGE C-2 Protocol418-014 Page 2 REGULATORY CITATIONS: U.S. Food and Drug Administration (1994). International Conference on Harmonisation; Guidelineon detectionof toxicittyo reproductionformedicinalproducts. Federal Register,September 22, 1994, Vol.59, No. 183. U.S. Food and Drug Administration.Good Laboratory PracticeRegulations;FinalRule. 21 CFR Part58. Japanese Ministryof Health and Welfare (1997). Good Laboratory Practice Standard forSafetyStudies on Dnigs, MHW Ordinance Number 21, March 26, 1997. European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decisionlrecommendation on compliance withprinciplesof good laboratorypractice.OfficiaJlournal of the European Communities: Legislation.32 (No. L 315; 28 October): 1-17. REGULATORY COMPLIANCE: This study willbe conducted incompliance with the Good Laboratory Practice(GLP) regulationscitedabove. Allchanges or revisionsof thisprotocolshallbe documented, signed by the Study Directorand the Sponsor, dated and maintained with the protocol. The QualityAssurance Unit(QAU) willauditthe protocol,the raw data and the report, and willinspectcriticaplhases of the study inaccordance with the Standard Operating Procedures of Argus Research Laboratories,Inc. The finalreportwillincludea statement signed by the Study Directorthatthe report accuratelyreflecttshe raw data obtained during the performance of the study and that allapplicableGLP regulationswere followed inthe conduct of the study. Should significandteviationsfrom GLP regulationsoccur, each willbe described indetail, togetherwithhow the deviationmight affectthe qualityor integritoyf the study. SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE: See ATTACHMENT 1 to the protocol. 418-014:PAGE C-3 Protocol418-014 Page 3 TEST ARTICLE AND VEHICLE: Identification: Test Article: Name: PhysicalDescription: Lot/BatchNumber SpecificGravity: Purity: ExpirationDate: PFOS (Synonym: FC-95). Light-coloredpowder. 217. -0.6. 98.9%. May 2000. Informationon the identityc,omposition,strengthand purityof the testarticleison file withthe Sponsor. Vehicle: 0.5% Tween@ 80 in Reverse Osmosis Membrane Processed Deionized Water (R.O. Deionized Water). Supplier and lotidentificatioofnTween@ 80 to be documented in the raw data. Neitherthe Sponsor nor the Study Directorisaware of any potentialcontaminants likely to be present inthe vehiclecomponents thatwould interferewiththe resultsof this study. Therefore,no analyses otherthan those mentioned inthisprotocolwillbe conducted. Safety Precautions: Gloves, mask, appropriateeye protectionand a unifonrdlabcoat are to be wom during formulationpreparationand administration.The Matedal Safety Data Sheet (MSDS) is attachedtothe protocol(ATTACHMENT 2). Stora-ge: Bulk Test Article: VehicleComponents: Prepared Vehicle: Prepared Formulations: Room Room Room Room temperature. temperature. temperature. temperature. 418-014:PAGE C-4 Protocol418-014 Page 4 Alltestarticleshipments to the Testing Facilitsyhould be addressed to the attentionof JulianGulbinski,Manager of Formulations,at the previouslycitedaddress and telephone number. Shipments should includeinformationconcerning storage conditionsand shipping cartonsshould be labeled appropriately.The recipientshould be notifiedin advance of shipment. FORMULATION: Freciuency of Preparation: Formulations(suspensions)willbe prepared dailyatthe Testing FacilityD.ata verifying the stabilitoyf the test articleinthe vehiclefor48 hours under the conditionsof administrationare on filewith the Sponsor. Detailedpreparationprocedures are attached to thisprotocol(ATTACHMENT 3). Adaustment for Purily: The testarticlweillbe considered 100% pure forthe purpose of dosage calculations. Testing FacilityReserve Samples: The Sponsor willreserve a sample (1 g) of each lotof the bulktestarticleused during the course ofthisstudy. The Testing Facilitwyillreserve a sample (5 mL) ofeach lotof the vehiclecomponents used during the course of thisstudy. Samples willbe stored under the previouslycitedconditions. ANALYSES: Samples additionalto those described below may be taken ffdeemed necessary during the course of the study. Bulk Test Article Samr)ling: No analyses of the bulk testarticlweillbe conducted during the course of thisstudy. Informationon the stabilitoyf the bulk testarticlieson filewiththe Sponsor. Analyses of Prepared Formulations: Homogeneity and stabilitoyf prepared formulationsison filewith the Sponsor. However, recordswillbe maintained to document how the testarticleformulationswere prepared. 418-014:PAGE C-5 Protor,o4l1M14 Page 5 ConcentrationofTest ArticlFeormulations: Concentration of the prepared formulationswillbe verifieduring the course of this study. Duplicatesamples (2 mL each) willbe taken from the firsatnd lastpreparation on the day prepared. One sample of each set willbe shipped foranalysis;the remaining samples willbe retainedat the Testing Facilitays backup samples. Backup samples willbe stored frozen (-700C or below) and discarded at the Testing Facility upon request of the Sponsor. Shipping Instructions: Samples to be analyzed willbe shipped (frozenon dry ice)to: Kds J. Hansen, Ph.D. 3M Environmental Technology and 935 Bush Avenue Building2-3E-09 St.Paul,Minnesota 55133-3331 Telephone: (612) 778-6018 Telefax: (612) 778-6176 Safety Services The recipientwillbe notifiedin advance of sample shipment. DISPOSITION: Prepared formulationswillbe discarded at the Testing FacilityA.llremaining bulk test articlweillbe returned to the Study Monitor at the previouslycitedaddress upon completion of allwork with the testarticle. TEST SYSTEM: Species/Strain and Reason for Selection: The Crl:CD@BR VAF/Plus@ (Sprague-Dawley) ratwas selected as the Test System because: 1) thisstrainof ratwas used inthe reproductiveand developmental toxicity studies;2) historicadlata and experience existatthe TestingFacilit,/'a-n'd);3)the test articleispharmacologicallyactiveinthe species and strain. 418-014:PAGEC-6 Protoco4l18-014 Page 6 Number: Initialpopulation acclimated: Population selectedforstudy: Population selectedfor pharmacokinetic sample collectio(nday 14 postpartum): Body Wei-cjhtand Age: 86 virginfemale rats. 54 mated female rats(30 inthe controlgroup and 24 inthe treatedgroup). 12 mated female rats(sixper dosage group). Female ratswillbe ordered tohave body weights of200 g to 225 g each at receipt,at which time they willbe expected to be at least60 days of age. Actual body weights will be recorded the day afterreceiptand willbe documented inthe raw data. The weight range willbe included inthe finalreport. Sex: Female ratswillbe given the testarticle.Male ratsof the same source and strainwill be used only as breeders and are not considered partof the Test System. Source: Chades River Laboratories,Inc. The ratswillbe shipped in filterecdartons by airfreightand/or truckfrom Charles River Laboratories,Inc.,to the Testing Facility. Identification: Fo Generation: Rats are permanently identifieudsing Monel@ self-piercinegar tags (Gey Band and Tag Co., Inc.,No. MSPT 20101). Male ratsare given unique permanent identification numbers upon assignment tothe Testing Facilit)b(rseeder male ratpopulation. Female ratsare assigned temporary numbers at receiptand given unique permanent identificationnumbers when assigned to the study. Fl Generation: Pups willnot be individualliydentifieddudng lactationa;llparameters willbe evaluated in terms of the lifter. 418-014:PAGE C-7 Protocol418-014 Page 7 ANIMAL HUSBANDRY: All cage sizes and housing conditions are in compliance with the Guide for the Care and Use of Laboratofy AnimaIS44@. Housina: Fo Generation Rats/Fl Generation Lifters: Fo generationratswillbe individuallhyoused in stainlesssteel,wire-boftomed cages, except duringthe cohabitationand postpartum periods. During cohabitation,each pair of ratswillbe housed inthe male rat'scage. Beginning no laterthan day 20 of presumed gestation,Fo generation female ratswillbe individuallhyoused in nesting boxes, except during collectionintervalsforurineand fecalsamples. During these collectionintervalst,he female ratswillbe housed individualliyn metabolism cages. Each dam and assigned littewrillbe housed in a common nesting box during the postpartum period. Nesting Material: Bedding material(bed-o'cobs(D)willbe provided. Bedding willbe changed as oftenas necessary to keep the animals dry and clean. Analyses forpossiblecontamination are conducted annuallyand documented inthe raw data. Room Air,Temperature and Humidi The animal room isindependently suppliedwith at leastten changes per hour of 100% freshairthathas been passed through 99.97% HEPA filter(sAiroClean@ room). Room temperature willbe maintained at640F (180C) to 79OF (260C) and monitored constantly.Room humiditywillalso be monitored constantlyand maintained at 30% to 70%. ligblt: An automaticallycontrolled12-hour light1:2-hour dark fluorescentlightcycle willbe maintained. Each dark periodwillbegin at 1900 hours EST. Diet: Rats willbe given CertifiedRodent Diet@ #5002 (PMI NutNtion Internationala)vailable ad libitumfrom individuafleeders. 418-014:PAGE C-8 Pmtocol 418-014 Page 8 Water: Water willbe available ad libitum from individualboftles attached to the cages or from an automatic watering access system. Allwater willbe from a localsource and passed through a reverse osmosis membrane before use. Chlorinewillbe added to the processed water as a bactedostat;processed water isexpected to contain no more than 1.2 ppm chlorineat the time of analysis.Water isanalyzed monthly forpossible bacterialcontamination and twiceannually forpossiblechemical contaminabon. Contaminants: Neitherthe Sponsor nor the Study Directorisaware of any potentialcontaminants likely to be present inthe certifiediet,the drinkingwater or the nesting materialat levelsthat would interferweith the resultsof thisstudy. Therefore,no analyses other than those routinelyperformed by the feed supplieror those mentioned in thisprotocolwillbe conducted. RANDOMIZATION, COHABITATION AND CROSS-FOSTERING PROCEDURES: Upon arrivalm,ale and female ratswillbe assigned to individualhousing on the basis of computer-generated random units.Afteracclimation,78 virginfemale ratswillbe selectedforstudy on the basis of physicalappearance and body weights recorded during acclimation.The female ratswillbe assigned to dosage groups (42 inthe controlgroup and 36 inthe treatedgroup) based on computer-generated (weightordered) randomization procedures and treatedwitheitherthe vehicleor the testarticle for42 days priorto cohabitation. Within each dosage group, consecutive order willbe used to assign female ratsto cohabitationwith breeder male rats,one male ratper female rat.The ratsassigned to the controlgroup willbe cohabited one day priorto cohabitationforratsassigned to the treatedgroup. The cohabitationperiodwillconsistof a maximum of fivedays. Female ratswith spennatozoa observed in a smear of the vaginalcontents and/or a copulatory plug observed in situwillbe considered to be at day 0 of presumed gestationand assigned to individualhousing. Mated female rats(those with confirmed evidence of mating)willbe assigned as follows:of the 42 female ratsassigned to the controlgroup, 30 mated female ratswill be assigned to the controlgroup forcross-fostedngpurposes; of the 36 female rats assigned to the treatedgroup, 24 mated female ratswillbe assigned to the treated group forcross-fostedngpurposes. An additionalsixmated female ratsfrom each dosage group willbe designated forpharmacokinetic sample collection(as described below) followingdelivery,but these ratswillnot be cmss-fostered. Finallya,ny remaining female ratswith confirmed evidence of mating thatwere assigned to the controlgroup priorto cohabitationwillbe assigned to the controlpool forcmss-fostering 418-014:PAGE C-9 Protocol418-014 Page 9 purposes. Any remaining female ratswith confirmed evidence of mating thatwerc@, assigned to the treatedgroup priorto cohabitationwillbe sacrificedand discarded withoutfurtherevaluationat the discretionof the Study Directorand the Study Monitor. Day 1 of lactation(postpartum) isdefinedas the day of birthand isalso the firstday on which allpups in a IKterare individuallwyeighed (pup body weights willbe recorded afterallpups in a littearre deliveredand groomed by the dam). Twelve dams and theirrespectivelitterwsillbe selectedand assigned to the pharmacokineticsample collectio(nsixdams and litterpser dosage group). These litterwsillnot be cross-fostered. Up to 24 lifterpser dosage group willbe reassigned forcross-fosteringon day 1 postpartum. The litterwsillbe assigned such thatfour subsets are obtained among the two dosage groups as described inthe table below. Dosage Group 1 1 It 11 Nurnber of Mated Female Rats Dosage Number of (mgvkgtday) UttersReassi ned Reassigned UtterType (Dam Treatment) 12 0 (Vehicle) 12 12 0 (Vehicle) 12 12 1.6 12 Treated Untreated Treated 12 1.6 1 12 1 Untreated Subset Number A B c D Detailedprocedures forreassignment of lifte6forcross-fostedngare attached to this protocol(ATTACHMENT 4). On day 1 postpartum, pups from dams assigned to the controlpool willbe culled, sacdficedvia decapitationand handled as described inATTACHMENT 4. On day 4 postpartum, cross-fosteredlifterwsillbe culledto fivemale and fivefemale pups per liftewrh,ere possible. Pups not selectedforcontinued evaluationwillbe sacdficedviadecapitation;the lungs (saved inBouin's solutiona)nd the liver(saved in neutralbuffered10% formalin)willbe collectedfrom the firsten culledpups from each dosage group (irrespectivoef liftedre)termined to be atday 4 postpartum and preserved forpossiblefuturehistopathologicaelvaluation.Remaining culledpups will be sacrificevdia decapitationand discardedwithout necropsy evaluation. 418-014:PAGE C-10 Protocol418-014 Page 10 ADMINISTRATION: Route and Reason for Choice: The oral(gavage) route was selectedforuse because: 1)thiswas the routeof administrationin the developmental and reproductivetoxicologystudies;and 2) itisone of the possibleroutes of human exposure. Method and Frequency: Fo Generation Female Rats: Female ratswillbe given the testarticleor the vehicleonce dailybeginning 42 days priorto cohabitationthrough day 21 postpartum. Dosages willbe adjusted dailyfor body weight changes and given at approximately the same time each day. Fl Generation Pups: Fl generation pups willnot be directlygiven the testarticleb,ut may be possibly exposed to the testarticleduring matemal gestation(inuteroexposure) or via matemal milkduring the lactationperiod. Rationale for Dosage Selection: Dosages were selectedon the basis of a previous study conducted with the testarticle (Argus Research Laboratories,Inc.,Protocol418-008). Dosage Levels,Concentrations and Volumes: Dosage Number ofMated Dosage Group FemaleRats (-g/k day) 1 30+61 0 (Vehicle) 11 24+61 1.6 Concentrabon (mg/mL) 0 0.32 Dosage Volume (mL/kg) ArgusBatchNumber 5 B-418-014-A(Day.Mn 0 -YYre)., 5 1 B:i@@ nth" The testarticlewillbe considered 100% pure forthe purpose of dosage calculations. a. Samples foranalysis of pharmacokinetics willbe collected from sixdams and #*r respecbve litterosn day 14 postpartum. These dams and tieirrespective litterwsillbe sac7ificW followingsample collection. TESTS, ANALYSES AND MEASUREMENTS - Fo GENERATION: Viability: AllPeriods: At leasttwice daily. 418-014:PAGE C-1 1 Protocol418-014 Page 11 Clinical Observations and/or General Appearance: Acclimation Period: At leastonce. Dosage Period: Twice daily.Priortoadministratioannd once approximatelyone hour postdosage. Matemal Behavior Days 1,4,7, 14 and 21 postpartum.Observed abnormal behaviorwillbe recordeddaily. Clinicaolbservationsmay be recorded more frequentlythan citedabove, ifdeemed appropriateby the Study Directorand/or Study Monitor. Body Weiahts: AcclimationPeriod: At leastonce. Dosage Period: Daily. Sacrifice: Terminal weight. Feed Consumption Values (recordedand tabulated): AcclimationPeriod: At leastonce. Dosage Period: Weekly to cohabitation.Dailyduringpresumed gestationand days 1,4, 7,10 and 14 postpartum. Feed consumption willnotbe tabulatedafterday 14 postpartum,when itisexpected thatpups willbegin to consume matemal feed. Feed consumption values may be recorded more frequentlyifitisnecessary to replenishthefeed. These intervalwsillnot be tabulated. Matin-qPerformance: Mating willbe evaluateddailyduringthe cohabitationperiodand confirmedby observationofspermatozoa ina smear ofthe vaginalcontentsand/ora copulatoryplug observed insitu. 418-014:PAGEC-12 Protocol418-014 Page 12 NaturalDelivery: Female rats willbe evaluated for ClinicaOlbservations During Parturitio[ntobe performed under red light conditionsduringthe dark period(timeeach pup isobserved willbe recorded)]. Durationof Gestation (day 0 of presumed gestationto the time the firsptup is observed). Length of Parturitio(ntimeof observationof lastpup minus the time of observationof the firstpup dividedby N-1 pups in each lifter). LifteSrize (definedas allpups delivered). Pup Viabilitayt Birth. Pharmacokinetic Sample Collection: Caps and labeledtubes willbe weighed (combined weight,to the nearest.001 gram) before and afterretentionof pooled pup samples forsubsequent use in pharmacokinetic analyses. These weights willbe documented inthe raw data,and copies of these weights willbe includedwith the packing listpriorto shipment. Blood, Milk and LiverSamr)les - Fo Generation Rats: Blood, milkand liversamples willbe collectedfrom sixmatemal ratsdesignated for phannacokinetic sample collectionper dosage group on day 14 postpartum. The time of sample collections(bloodand milk)willbe recorded inthe raw data. Milk samples willbe collectedapproximately two to sixhours postdosage on day 14 postpartum. Each dam willbe removed from the nestingbox and individuallhyoused forapproximatelyfour hours. The dam willbe injectedintravenouslywith one unitof oxytocinapproximately fiveminutes before milksamples (atleast100 pL per sample) are collected.The samples willbe immediately frozen on dry iceand maintained frozen (-70*C orbelow) untilshipment to the Sponsor foranalysis. Followingmilksample collectionb,lood samples (approximately4 mL each) willbe collectedfrom the matemal ratsviathe inferiovrena cava and transferredintoserum separatortubes. The samples willbe spun in a refrigeratecdentrifuge.The serum will be transferredintopolypropylene tubes labeledwith the study number, animal identificatiodna,te of collections,tudy day and collectiotnimepoint.Allsamples willbe immediatelyfrozen on dry iceand maintained frozen (-700C or below) untilshipment to the Sponsor foranalysis. 418-014:PAGE C-1 3 Protocol418-014 Page 13 Followingcollectioonf milkand bloodsamples,a liversection(righltaterallobe)and the milk-secretinglands from the axillaryt,horacic,abdominal and inguinalregionsoi each dam (lefstide only)willbe collected,frozenand stored(-700C or below) untilshipment to the Sponsor foranalysis. Blood and LiverSamples - Fl Generation Pur)s: Blood samples willbe collectedviathe inferiovrena cava from each pup on day 14 postpartum, pooled (per liftera)nd transferredintoserum separatortubes. The samples willbe spun in a refrigeratecdentrifuge.The serum willbe transferredinto polypropylene tubes labeledwith the study number, animal identificatiodna,te of collections,tudy day and collectiotnimepoint.Allsamples willbe immediately frozen on dry iceand maintained frozen (-700C or below) untilshipment tothe Sponsor for analysis. The liverfrom each pup willbe collected,pooled (perlifterf)r,ozenand stored(-700C or below) untilshipment to the Sponsor foranalysis. Shipping Instructions: Allsamples willbe maintained frozen(-70'C or below) untilshipment foranalysis. Samples willbe shipped on frozen on dry icevia overnightmail.A packing liswtillbe includedwith the samples and sent to KrisJ. Hansen, Ph.D., at the previouslycited address. Both the recipientand the Study Monitorwillbe notifieidn advance of sample shipment. Urine and Fecal Sample Collection: Fo generation female ratswillbe housed individualliyn metabolism cages for collection of urineand fecalsamples from days 21 to 22 postpartum. Followingthe 24-hour collectionintervals,amples willbe collectedintocentrifugetubes, placed on dry ice and storedfrozen(-70'C or below) untilshipment foranalysis. Shipping Instructions: Allsamples willbe maintained frozen(-70'C or below) untilshipment foranalysis. Samples willbe shipped on frozen on dry iceviaovernightmail. A packing liswtillbe includedwith the samples and sent to KrisJ. Hansen, Ph.D., at the previouslycited address. Both the recipientand the Study Monitorwillbe notifiedinadvance of sample shipment. METHOD OF SACRIFICE - Fo GENERATION RATS: Rats willbe sacrificedby carbon dioxide asphyxiation. 418-014:PAGE C-14 Protocol418-014 Page 14 NECROPSY -Pa GENERATION RATS: Gross lesions will be retained in neutral buffered 10% formalin for possible future evaluation (a table of random unitswillbe used to selectone controlgroup ratfrom which alltissues examined at necropsy willbe retained,inorder to provide control tissuesfor any possible histopathologicalevaluations of gross lesions).Unless specificalcliytedbelow,allothertissueswillbe discarded. Rats Not Deliverin-q a Lifte Rats that do not deliver a litterwill be sacrificed on day 25 of presumed examined forgross lesions. Uteriwillbe stained with 10% ammonium the absence of implantationsftes(5). gestation and sulfideto confirm Scheduled Sacirifice- Rats Assi-qned to Pharmacokinetic Samg)le Collection: On day 14 postpartum, milk and blood samples, a liversection (rightlaterallobe) and the milk-secretingglands from the axillaryt,horacic,abdominal and inguinalregions of each dam (leftside only)willbe collectedas described previously,frozen and stored (-700C or below) untilshipment to the Sponsor foranalysis. Following the completion of pharmacokinetic sample collection,Fo generation female ratswillbe sacrificed,and a gross necropsy of the thoracic,abdominal and pelvicviscera willbe performed. The number and distributionof implantation siteswillbe recorded. Scheduled Sacrifice - Day 22 Postpartum: On day 22 postpartum, blood samples willbe collectedfrom the Fo generation female ratsfollowingremoval from metabolism caging. Blood samples (approximately 4 mL each) willbe collectedfrom the matemal rats via the inferiovrena cava and transferredintoserum separator tubes. The samples willbe spun ina refrigeratedcentrifuge. The serum willbe transferredintopolypropylene tubes labeled with the study number, animal identificationd,ate of collection,study day and collectiontimepoint. Allsamples willbe immediately frozen on dry ice and maintained frozen (-700C or below) untilshipment to the Sponsor for analysis. The female ratswillthen be sacrificed,and a gross necropsy of the thoracic,abdominal and pelvicviscera willbe performed. The number and distributioonf implantation sites willbe recorded. A liversection (rightlaterallobe) from each dam willbe collected, frozen and stored (-700C or below) untilshipment to the Sponsor for analysis. 418-014:PAGE C-15 Protocol418-014 Page 15 Dams with No SurvivinciPups: Dams with no surviving pups willbe sacrificed afterthe lastpup isfound dead, missing or presumed cannibalized. Priorto sacrificea, blood sample (approximately4 mL) willbe collectedfrom the matemal ratviathe inferiovrena cava and transferredintoserum separatortubes. The sample willbe spun in a refrigeratecdentrifuge.The serum willbe transferredintoa polypropylene tube labeledwith the study number, animal identificatiodna,te of collections,tudy day and collectiotnimepoint. The sample willbe immediately frozen on dry iceand maintained frozen (-700C or below) untilshipment to the Sponsor for analysis. A gross necropsy of the thoracic,abdominal and pelvicviscerawillbe performed. A liversection(rightlateralobe)from each dam willbe collected,frozen and stored (-700C or below) untilshipment to the Sponsor foranalysis.Postpartum data forthese dams willbe excluded from summary tables. Rats Found Dead or Moribund: Rats thatdie or are sacrificedbecause of moribund condition,abortionor premature deliverywillbe examined forthe cause of death or moribund conditionon the day the observationismade. The ratswillbe examined forgross lesions.A liversection(right lateralobe)from each dam willbe collected,frozenand stored(-700C or below) until shipment to the Sponsor foranalysis. Pregnancy statusand uterinecontents of female ratswillbe recorded. Aborted fetuses and/or deliveredpups willbe examined to the extent possible.Uteriof apparently nonpregnant ratswillbe stainedwith 10% ammonium sulfideto confin-nthe absence of implantationsiteS(5). Shipping Instructions: Allsamples willbe maintained frozen (-70*C or below) untilshipment foranalysis. Samples willbe shipped frozen on dry ice viaovernightmail. A packing liswtillbe includedwith the samples and sent to KrisJ. Hansen, Ph.D., at the previouslycited address. Both the recipientand the Study Monitor willbe notifiedinadvance of sample shipment. TESTS, ANALYSES AND MEASUREMENTS - Fl GENERATION: Viability: Postpartum Period: Litterswillbe observed fordead pups at leasttwice daily.The pups ineach liftewrillbe counted once daily. 41M14:PAGE C-16 Protor-o4l18-014 Page 16 Clinical Observations and/or General Appearance: Postpartum Period: Once daily. Clinicalobservationsmay be recorded more frequentlythan citedabove, ifdeemed appropriateby the Study Directorand/or the Study Monitor. Body Wei-ghts: Postpartum Period: Sacrifice: Days 1 (birth)4, 7, 14 and 21 postpartum. Terminal weight. METHOD OF SACRIFICE - Fl GENERATION RATS: As previouslycitedforFo generation rats.Culled pups willbe sacrificevdia decapitation. NECROPSY - Fl GENERATION RATS: Gross lesionswillbe retainedin neutralbuffered10% formalinforpossiblefuture evaluation(a tableof random unitswillbe used to selectone controlgroup ratof each sex from which alltissuesexamined at necropsy willbe retained,in order to provide controltissuesforany possiblehistopathologicaelvaluationsof gross lesions).Unless specificallcyitedbelow, allothertissues willbe discarded. As described previously,caps and labeled tubes willbe weighed (combined weights, to the nearest.001 gram) before and afterretentionofpooled pup samples. These weights willbe documented inthe raw data,and copies of these weights willbe includedwith the packing listpriorto shipment. Pups Found Dead on Day 1 Postpartum.: Pups thatdie before examination of the liftefrorpup viabilitwyillbe evaluated forvital statusat birth.The lungs willbe removed and immersed inwater. Pups with lungs that sinkwillbe identifieads stillbom;pups with lungs thatfloatwillbe identifieads livebom, and to have died shortlyafterbirth.Pups with gross lesionswillbe preserved in Bouin's solutionforpossiblefutureevaluation.The lungs willbe preserved in Bouin'ssolution and the liverwillbe preserved in neutralbuffered10% formalinforpossiblefuture evaluation. 418-014: PAGE C-1 7 Protocol4lMl4 Page 17 Pups Found Dead or Moribund on Days 2 to 21 POStRartum: Pups found dead or sacrificed because of moribundity willbe examined for gross lesionsand forthe cause of death or the moribund condition.Pups with gross lesions found on days 2 to 4 postpartum willbe preserved in Bouin'ssolutionforpossiblefuture evaluation;gross lesionsof pups found on days 5 to 21 postpartum willbe preserved in neutralbuffered10% formalin. For allpups found dead on days 2 to 4 postpartum, the lungs willbe preserved in Bouin'ssolutionand the liverwillbe preserved in neutralbuffered 10% formalinfor possiblefutureevaluation.For allpups found dead on days 5 to 21 postpartum, the lungs and the liverwillbe preserved in neutralbuffered10% formalinforpossiblefuture evaluation. Pups Not Selected for Continued Observation - Days I and 4 Postg)artum: Allpups culledon day 1 postpartum (pups from dams assigned to the controlpool)and day 4 postpartum (allother culledpups) willbe sacrificedviadecapitation.The lungs and the liverwillbe collectedfrom the firsten pups culled(irrespectivoef liftefrr)om each dosage group on each day (controlpool only on day 1 postpartum);the lungs will be retainedinBouin'ssolution,and the liverswillbe retainedinneutralbuffered 10% formalinforpossiblefutureevaluation.Remaining culledpups willbe sacrificedand discarded withoutevaluation. Scheduled Sacrifice- LittersAssigned to Pharmacokinetic Sample Collection: On day 14 postpartum, lifterasssigned to the pharmacokinetic sample collectiownillbe sacrificewdiththeirrespectivedams. Individualp,ooled samples (perlifteorf) blood and liverwillcollectedas described previously.The pups willbe examined forgross lesions. Scheduled Sacrifice- LittersAssigned to Sacrificeon Day 21 Postpartum,: On day 21 postpartum, six lifterpser subset willbe randomly selected forcollectioonf pooled samples (perliftero)f blood and liver. Blood samples willbe collectedviathe inferiovrena cava from each pup, pooled (per liftera)nd transferredintoserum separator tubes. The samples willbe spun ina refrigeratecdentrifuge.The serum willbe transferredintopolypropylene tubes labeled withthe study number, animal identificatiodna,te of collections,tudy day and collection timepoint.Allsamples willbe immediately frozen on dry iceand maintained frozen (-70'C or below) untilshipment tothe Sponsor foranalysis. 418-014:PAGE C-18 Protocol418-014 Page 18 The pups willbe examined forgross lesions.The livefrrom each pup willbe collected, pooled (perlitterf)r,ozen and stored(-70'C or below) untilshipment tothe Sponsor for analysis. Allotherremaining pups willbe sacrificedand discarded withoutfurtherevaluation. ShiPRin-qInstructions: Allsamples willbe maintained frozen (-70*C or below) untilshipment foranalysis. Samples willbe shipped frozen on dry iceviaovernightmail. A packing liswtillbe includedwith the samples and sent to KrisJ. Hansen, Ph.D., at the previouslycited address. Both the recipientand the Study Monitorwillbe notifiedinadvance of sample shipment. STATISTICAL EVALUATION: Averages and percentages willbe calculated.Liftervalues willbe used where appropriate.Additionalprocedures and/or analyses may be performed, ifdeemed appropriate. DATA ACQUISITION, VERIFICATION AND STORAGE: Data willbe hand- and/or computer-recorded. Records willbe reviewed by the Study Directorand/or appropriatemanagement personnel within21 days aftergeneration. All originalrecords willbe stored inthe archivesof the Testing Facility.Alloriginadlata will be bound and indexed. A copy of allraw data willbe supplied to the Sponsor upon request. Preserved tissueswillbe stored at the Testing Facilitayt no charge forone year aftermailingof the draftfinalreport,afterwhich time the Sponsor willbe contacted to determine the dispositionof these materials. 418-014:PAGE C-19 Protocol41" 14 Page 19 RECORDS TO BE MAINTAINED: Protocol and Amendments. Test ArticleV,ehicle and/or Reagent Receipt,Preparation and Use. Animal Acquisition. Randomization Schedules. Mating History. Treatment (ifprescribedby StaffVeterinarian). General Comments. ClinicalObservations and/or General Appearance. Tissue and Sample Collection,Processing and Shipment. Cap and Labeled Tube Weights. Body Weights. Feed Consumption Values. NaturalDeliveryObservations. LitteOrbservations. Gross Necropsy Observations. Organ Weights (ifrequired). Photographs (ifrequired). Study Maintenance (room and environmentalrecords). Feed, Water and Bedding Analyses. Packing and/or Shipment Lists. KEYPERSONNEL: ExecutiveDirectorof Research: MildredS. Christian,Ph.D., Fellow,ATS Directorof Research: Alan M. Hoberman, Ph.D., DABT Associate Directorof Research and Study Director Raymond G. York, Ph.D., DABT Directorof Laboratory Operations: John F. Barnett,B.S. Manager of Study Coordination: ValerieA. Sharper, M.S. Manager of Animal Operations and Member, InstitutionAanlimal Care and Use Committee: Dena C. Lebo, V.M.D. Manager of Regulatory Compliance: Barbara J. Patterson,B.A. Consultant,VeterinaryPathology: W. Ray Brown, D.V.M., Ph.D.,ACVP 418-014:PAGE C;-20 Pmtocol4lMl4 Page 20 FINAL REPORT: A comprehensive draftfinalreport willbe prepared on completion of the study and will be finalizedfollowingconsultationwith the Sponsor. The reportwillincludethe following: Summary and Conclusion. Experimental Design and Method. Evaluationof Test Results. Appendices: Figures,Summary and IndividuaTlables Summarizing the Above Data, Protocoland Associated Amendments and Deviations,Study Directoes GLP Compliance Statement, Reports of SupportingData (ifappmpdate) and QAU Statement. INSTITUTIONAL ANIMAL CARE AND USE COMMITTEE STATEMENT: The procedures described inthisprotocolhave been reviewed by the Testing Facilitys InstitutionAanlimal Care and Use Committee. Allprocedures described inthisprotocol thatinvolvestudy animals willbe conducted in a manner to avoid or minimize discomfort,distressor pain to the animals. The Sponsoes signaturebelow documents the factthatinformationconcerning the necessityforconducting thisstudy and the factthatthisisnot an unnecessarily duplicativestudy may be obtained from the Sponsor. No alternative(invitro) procedures were availableformeeting the stated purposes of the study. .418-014:PAGE C-21 Protocol418-014 Page 21 REFERENCES: 1 Christian, M.S. and Voytek, P.E. (1982). In Vivo Reproductive and Mutagenicity Tests. Envimnmental Protection Agency, Washington, D.C. National Technical Information Service, U.S. Department of Commerce, SpringfieldV,A 22161. 2. Christian,M.S. (1984). Reproductive toxicityand teratologyevaluations of naftrexone(Pmceedings ofNaftrexoneSymposium, New York Academy of Sciences, November 7, 1983), J. Clin.Psychiat.45(9):7-10. 3. Lang, P.L. (1988). Embryo and Fetal Developmental Toxicity(Teratology) Control Data in the Chartes River Crt.CDOBR Rat. Charles River Laboratories, Inc.,Wilmington, MA 01887-0630. (Database provided byargus Research Laboratories,Inc.) 4. Instituteof Laboratory Animal Resources (1996). Guide forthe Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 5. Salewski, E. (1964). F5rbemethode zum makroskopischen Nachweis von lmplantationsstellenam Uterus der Ratte. Arch. Pathol. Exp. Pharmakol. 247:367. PROTOCOL APPROVAL: FOR THE TESTING FACILITY 0- 4r, /- Alan M. Hoberman, Ph.D.,DABT Directorof Research ond G. York,@h@., DABT AssociateDirectorof Research Study Director Dena C. Lebo,V.M.D. Member, InstitutioAnnailmal Care and Use Committee FOR THE SPONSOR MarvinT. Case, D.V.M.,Ph.D. Study Monitor 418-014:PAGE C-22 Protocol418-014 Page 22 2 -2 Date -f-1':F 7-3- Oc--r-9 f3 Date 9-Z Date Date 418-014:PAGE C-23 ATTACHMENT I SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE ATTACHMENT I 418-014:PAGE C-24 Protocol418-014 Page 1 of 2 STUDY SCHEMATIC CROSS-FOSTERING STUDY8 Staqof Dosage Femal Rat Premabng Pedod (42Dap) Endof Dosage ScheduW Saoifcdel, Cohabbton Pe@@ (5dap) Presumed Gestbon Pe@od Pos@atm/ Ladaton Pe@odb Dosage Period. a. For additionaldetailssee "Tests,Analyses and Measurements" sectionof the protocol. b. Fl generationpups willbe cross-fosterebdeginningon day 1 postpartumand continuinguntilday 21 postpartum. C. Fl generation pups willbe sacrificedon day 21 postpartum. Fo generationdams willbe sacrificedon day 22 postpartum. d. Samples foranalysisof pharmacokinetics willbe collectedfrom sixdams and their respectivelifterosn day 14 postpartum. These lifterwsillnot be cross-fosteredand willbe sacrificedfollowingsample collection. 418-014:PAGEC-25 ATTACHMENT 1 Pmtocol 418-014 Page 2 af2 27 OCT 98 02 NOV 98 SCHEDULER Animals Anive - AcclimationBegins. Dosage Period - Female Rats (42 days priorto cohabitationuntilday 21 postpartum). 13 DEC 98 PM - 19 DEC 14 DEC 98 19 DEC 98 98 AM 04 JAN 99 13 JAN 99 04 JAN 99 12 JAN 99 08 JAN 99 16 JAN 99 08 JAN 99 13 JAN 99 17 JAN 99 26 JAN 99 24 JAN 99 - 02 FEB 99 25 JAN 99 - 03 FEB 99 Cohabitation Period. FirstPossible Day 0 of Presumed Last Possible Day 0 of Presumed Gestation. Gestation. FirstPossible Delivery(Day 21 of presumed gestation). Last Possible Delivery(Day 25 of presumed gestation). FirstPossible Day 1 Postpartum CullingControl Pool Only. Last Possible Day 1 Postpartum CullingControl Pool Only. FirstPossible Day 4 Postpartum CullingCross-Fostered Litters. Last PossibleDay 4 Postpartum CullingCross-Fostered Litters. FirstPossible Day 25 of Presumed Female Sacrifice. Last Possible Day 25 of Presumed Female Sacrifice. Gestation Gestation FirstPossible Day 14 Postpartum Pharmacokinetic Sacrifice. Last Possible Day 14 Postpartum Pharmacokinetic Sacrifice. Scheduled Sacrifice- Fl Generation Pups (Day 21 postpartum). Scheduled Sacrifice- Fo Generation Dams (Day 22 postpartum). 08 JUN 99 DraftFinalReport. a. The study initiatiodnate isthe day the Study Directorsignsthe protocol. 4'18-014:PAGE C-26 ATTACHMENT2 MATERIAL SAFETY DATA SHEET 418-014-.PAGE C--/-7 MATERIAL SAFETY DATA SKUT 3H 3H Center st. Paul, Minnesota SSI"-1000 1-600-364-3S77 or (612) 737-SSOI (24 hours) CDpyright, igge, Minnesota Mining and Manufacturing Company. All rights reserved. Copying and/or downloading of this information for the purpose of properly utilizing 314products is allowed provided that: 1) th* information is copied in full with no changes unless prior agreement is obtained from 3M, and 2) neither the copy nor the original is resold or otherwise distributed with the intention of earning a profit thereon. IDIVISION: 3H CHEMICALS -MADE NAME: FC-96 FLUDW Brand Fluorochomical Surfactant ID NUMBER/U.P.C.: 98-0207-0103-7 00-51135-09064-1 98-0207-0104-5 98-0211-0888-5 ZF-0002-10"-l 00-SI13S-09362-7 - 98-0211-3916-1 ISSUED: January 29, 1998 SUPERSEDES: November 05, 1997 IDOCUMENT: 10-3796-9 00-5113S-09OSS-8 00-51135-02311-2 ----------------------------------------------------------------------------- 1 . INGREDIENT C.A.S. NO. PERCENT ----------------------------------------------------------------------------- POTASSIUM POTASSIUM POTASSIUM POTASSIUM POTASSIUM PERFLUOROALKYL PERFLUOROALKYL PERFLUOROALKYL PERFLUOROALKYL PERFLUOROALKYL SULFONATE ...... 279S-39-3 82 SULFORATE ...... 3871-99-6 3 SULFONATE ...... 29420-49-3 3 SULFONATE ...... 60270-S5-5 2 SULFONATE ...... 3872-2S-1 1 - as .a -7 .6 -3 ----------------------------------------------------------------------------- 2. PHYSICAL DATA ----------------------------------------------------------------------------- BOILING POINT: ...............N./.A VAPOR PRESSURE: ..............N./.A VAPOR DENsrTY: ...............N./.A EVAPORATION RATE: .............N/.A SOLUBILITY IN WATER: ..........s.light SPECIFIC GRAvrTY: .............ca.. 0.6 Water-I PERCENT (Bulk) VOLATILE: .............0.% PH: ........................7..-.6. (0.1% Aqueous) VISCOSITY: ..................N./.D. MELTING POINT: ...............N./.D A-PPEARANCE MD ODOR: Light colored, free flowing powder. - ---------------------------------------------------------------------------A.bbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 418-014:PAGE C-28 MMS: Fr,.95FLUORAD Brand FluorochesUal Surfactant January 29, 1998 PAGE 2 ------------------------------------------------------------------------- 3. FIRE AND EXPLOSroN mAzARD DATA ------------------------------------------------------------------------- FLASH POINT:...............N.on.e.. FLAM4ABLE LIMITS - LF-L:.......N./A FLAMMABLE LIMITS - UEL: .......N./A AuToiGNrTION TEMPERATURE: ......N/A EXTINGUISHING MEDIA: Water, Carbon dioxide, Dry chemical, Foam SPECIAL FIRE FIGHTING PROCEDURES: Wear full protectiveclothing,includinghaimt, self-contained, positive pressure or pressure demand breathing apparatus,bunker coat and pants,bands around arms, maint and logs, face mask, and protectivecoveringfor exposed areas of the head. UNUSUAL FIRE AND EXPLOSION HAZARDS: See hazardousDecomposition section for products of combustion. ------------------------------------------------------------------------4. REACTIVITY DATA ------------------------------------------------------------------------- STABILITY: Stable INCOMPATIBILITY - MATERIALS /CONDITIONS TO AVOID: Not applicable. HAZARDOUS POLYMERIZATIDN: Hazardous polymerization will not occur. HAZARDOUS OECOMPOSITION PRODUCTS: Carbon Monoxide and Carbon Dioxide, Oxides of Sulfur, Hydrogen Fluoride,Toxic Vapors, Gases or Particulates. ------------------------------------------------------------------------S. ENVIRONMENTAL INFORMATION ------------------------------------------------------------------------- SPILL RESPONSE: Observe precautionsfrom other sections. Vacuum, use wet sweeping compound or water to avoid dusting. CAUTIONI A vacuum cleaner could be an ignitionsource. Clean up residuewith water. Place in an approved metal container. Seal the container. REC"ENDF-D DISPOSAL: Do not release to waterways or sewer. Do not use in productsor processesthat could result in aqU2tiCconcentrationsgreater than 1/10 of the lowest EC50 or LCSO concentration. Incineratein an industrialor commercialfacilityin the presenceof a combustible material. Combustionproducts will includeHF. Disposal alternative: Disposeof waste productin a facilitypermittedto ------------------------------------------------------------------------Abbreviations:N/D - Not Determined N/A - Not Applicable CA - Approximately 418-014:PAGE C-29 "SOS: FC.gs prLuoRADBrand Fluorochenical Surfectant isnuary 29, 1998 PAGE 3 ------------------------------------------------------------------------5. EWIROWIMAL INFOR14ATION (continued) ------------------------------------------------------------------------- accept chemical waste. ENVIROWIMAL DATA: 96-Mr. Aquatic Fish LCSD, Fathead Kinnow(Pimephalesproselas)-38moil, BluegiU Suntish(Lopomismacrochirus)-W mg/l, Rainbow Trout(Salmo gairdneri)-Ilmg/l; 46-Mr. ECSO, DaphnisKagna a 50 ag/l; COD-.004 gig; B0020- Nil. REGULATORY INFORMATION: volatile Organic Compounds: N/A. VOC Less M20 & Exempt Solvents: N/A. Since regulationsvary, consult applicableregulationsor authorities before disposal. U.S. EPA Hazardous Waste Number- None (Not U.S. EPA liazardous). This productcomplieswith the chemicalregistrationrequirementsof TSCA, EINECS, CDSL, AICS, MITI and Korea. EPCRA K4ZARD CLASS: FIRE MAZARD: No PRESSURE: No REACTIVITY: No ACUTE: Yes CHRONIC: Yes ------------------------------------------------------------------------6. SUGGESTED FIRST AID ------------------------------------------------------------------------- EYE CONTACT: Immediatelyflush eyes with large amountsof water for at least 15 minutes.Get immediate medical attention. SKIN CONTACT: Immediatelyflush skin with large amounts of water. Remove contaminatedclothing.If irritationpersists,call a physician.Wash contaminatedclothing before reuse. INHALATION: If signs/symptomsoccur, remove person to fresh air. If signs/symptomscontinue, call a physician. IF SWALLOWED: Drink two glassesof water. Call a physician. ------------------------------------------------------------------------7. PRECALRTIONARY INFORMATION ------------------------------------------------------------------------- EYE PROTECTION: Avoid eye contact. Wear vented goggles. ------------------------------------------------------------------------Abbreviations:N/D - Not Determined N/A - Not Applicable CA - Approximately 418-014:PAGE C-30 NDS: FC-95FLUORAD Brand fluorochosical Surfactant January 29, 1998 PAGE 4 ------------------------------------------------------------------------- 7. PREcAuriomARY INFORPATION (continued) ------------------------------------------------------------------------------ SKIN PROTECTION: Avoid skin contact. Wear appropriategloveswhen handlingthis material. A pair of gloveamad* from the followingmaterial(s)are recommended: butyl rubber. Use one or more of the following personalprotectionitems as necessaryto prevent skin contact: head covering,coveralls. Protectivegarments(otherthan gloves)should be made of either of the followingmaterials: polyethylenep/olyvinylidenechloride (Saranex). RECOMMENDED VENTILATION: Use withappropriatelocal exhaustventilation. Use in a wallventilatedarea. Providesufficientventilatlonto maintain emissionsbelow recommended exposurelimits. If exhaust ventilation is not adequate,use appropriaterespiratoryprotection. RESPIRATORY PROTECTION: Avoid breathingof dust. Select one of the followingHIO&H approved respiratorsbased on airborne concentrationof contaminantsand in accordancewith OSHA regulations: half-maskdust and mist respirator, half-masksuppliedair respirator,full-facedust and mist respirator, full-facesuppliedair respirator. PREVENTION OF ACCIDENTAL INGESTION: Do not eat, drink or smoke when using this product. Wash exposed areas thoroughlywith soap and water. Wash hands after handling and before eating. RECOMMENDED STDRAGE: Keep containerdry. Keep container closed when not in use. FIRE AND EXPLOSION AVOIDANCE: Nonf lammable. OTHER PRECAUTIONARY INFORMATIDN: No smoking:Smoking while using this productcan result in contamin2tiOnOf the tobacco and/or smoke and lead to the formation of the hazardousdecomposition products mentionedin section 4 of this MSDS. HMIS HAZARD RATINGS: HEALTH: 2 FLAMMABILITY: 0 REACTIVITY: 0 PERSONAL PROTECTION: X (See precautions, section 7.) EXPOSURE LIMITS INGREDIENT VALUE UNIT TYPE AUTH SKIN* -------------------------------------------------------------------------- POTASSIUM PERFLUOROALKYL SULFONATE ... 0.1 MG/M3 TWA 3M y POTASSIUM PERFLUDROALKYL SULFONATE ... 0.1 MG/M3 TWA 3M y POTASSIUM PERFLUDROALKYL SULFONATE ... 0.1 MG/M3 TWA 3M y POTASSIUM PERFLUOROALKYL SULFONATE ... 0.1 MG/M3- TWA 3M y -------------------------------------------------------------------------- Abbreviations:N/D - Not Determined N/A - Not Applicable CA - Approximately Ml 4:PAGE C-3", MDS: FC.gs pLuoM Brand FluorochosiomaSurfactent January 29, IM PAGE 5 E)"SURE LIMITS (continued) INGREDIENT VALUE UNIT TYPE AUTH SKIN* -------------------------------------------------------------------------- POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 NGIN3 TWA 3H y * SKIN NOTATION: Listed substances indicated with 'Y' under SKIN refer to the potentialcontributionto the overallexposure by the cutaneousroute Including mucous membrane and eye, either by airborneor, more particularly, by direct contactwith the substance. Vehicles can alterskin absorption. SOURCE OF EXPOSURE LIMIT DATA: - 3M: 3H Recommended Exposure Guidelines -------------------------------------------------------------------------8. HEALTH HAZARD DATA -------------------------------------------------------------------------- EYE CONTACT: Mild Eye Irritation:signalaymptom3can include redness,swelling, pain, and tearing. SKI N CONTACT: Mild SkinIrritation(afterprolongedor repeatedcontact): signs/symptomscan include redness,swelling, and itching. May be absorbedthrough the skin and persist in the body for an extended time. INKALATION: May be harmful if inhaled. May be absorbed by inhalationand persist in the body for an extended time. Single overexposure, above recommended guidelines, may cause: Irritation(upper respiratory):signs/symptomscan include soreness of the nose and throat, coughing and sneezing. IF SWALLOWED: Ingestionis not a likelyroute of exposureto this product. Illnessmay result from a single swallowingof a moderatequantityof this material. May be harmful if swallowed. f4UTAGENICITY: Mutagenicityassays indicatethe productis not outagenic. -------------------------------------------------------------------------Abbreviations:N/D - Not Determined N/A - Not Applicable CA - Approximately 418-014:PAGE C-32 pu=: FC-96 FLUORAD Brand Fluorochosical Surfactant January 29, 1998 PAGE 6 ------------------------------------------------------------------------B. HEALTHHAZARDDATA (continued) ------------------------------------------------------------------------- REPRODUCTIVE/DEVELOPMENTAL TOXINS: Not toratogenicin the rat at oral done& belowmaternallytoxic levels. OTHER HEALTH HAZARD INFORMATION: This product is not known to contain any substancesregulatedunder CaliforniaProposition65. A Product Toxicity SummaryShoot is available. ------------------------------------------------------------------------SECTION CHANGE DATES ------------------------------------------------------------------------- HEADING SECTION CHANGED SINCE November 06, 1997 ISSUE ------------------------------------------------------------------------Abbreviations:N/D - Not Determined N/A - Not Applicable CA - Approximately ------------------------------------------------------------------------- The informationin thisMaterial Safety Data Sheet (MSDS)is believedto be correct as of the date issued. 3M MAKES NO WARRANTIES, EXPRESSED OR IMPLIED, INCLUDING, BUT NOT LIMITED TO, ANY IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE OR COURSE OF PERFORMANCE OR USAGE OF TRADE. User Is responsible for determining whether the 3M productis fit for a particularpurposeand suitablefor user's method of use or application. Giventhe varietyof factorsthat can affect the use and applicationof a 3H product,some of which are uniquelywithin the user'sknowledgeand control,it is essentialthat the user evaluatethe 3M product to determinewhetherit is fit for a particularpurposeand suitablefor user'smethodof use or application. 3M provides informationin electronicform as a serviceto its customers. Due to the remote possibilitythat electronictransfermay have resulted in errors, omissionsor alterationsin th13 information,3M makes no representationsas to its completenessor accuracy. In addition, informationobtainedfrom a database may not be as current as the informationin the MSDS available directly from 3M. 418-014:PAGE C-33 ATTACHMENT 3 TEST ARTICLE PREPARATION PROCEDURE 418-014:PAGE C-34 ATTACHMENT TEST ARTICLE PREPARATION Protocol418-014 Version:418-014 (22OCT 98) Page I of 2 PROCEDURE Test Article: PFOS. Vehicle: 0.5% Tween(D 80 in R.O. Deionized Water. A. Purpose: The purpose of thisprocedure isto providea method forthe preparation of dosage suspensions of PFOS and the vehiclefororaladministrationto ratson Argus Study 418-014. B. General Information: 1 . Allsuspension containerswillbe labeledand colorcoded. Each labelwill specifythe protocolnumber, testarticliedentificatioAnr,gus batch number, concentration,dosage level,preparationdate,expirationdate and storage conditions. 2. Suspensions willbe prepared: X Daily - Weekly - For- days ofuse 3. Vehicle willbe prepared: Daily X Weekly - For days of use 4. Suspensions willbe prepared at a finaldosage volume of 5 mukg. 5. Safety: X Gloves, lab coat,goggles or safetyglasses and faceshield x Dust-Mist Respirator - Half-Face Respirator - Full-Face Respirator/PositivPeressure Hood - Tyvek Suit/Apron 6. Dosage solutionsadjustedforFree base and % Purity. Yes X No (Calculationsbased on 100%) Free Base Purity 7. Sampling requirements: Cited in protocol. 8. Storage: Cited inprotocol. 418-014:PAGE C-35 ATTACHMENT 1 TEST ARTICLE PREPARATION Pmtocol 418-014 Version:418-014 (22 OCT 98) Page 2 of2 PROCEDURE NOTE: Once the finalvolumes are achieved,stirbarsare tobe added tothe containersm;ixingshouldoccurduringsamplingand/oradministration. C. PreparationofVehicle: 1 . Add the requiredamount ofR.O. deionizedwatertoan appropriately labeledcontainer.Heat thewaterto50"C, :t5'C,add the required amount ofTween(D 80 and mix untiulniform.(See TEST ARTICLE CALCULATIONS forexactquantities.) D. Test ArticlSeuspension Preparation: 1 To preparethe suspensionforthe highdosage group,add the required amount oftestarticl(eSee TEST ARTICLE CALCULATIONS) intoan appropriatelsyized,labeledcontainer.QS ad tothe requiredamount with vehiclea,dd a stirbar and heatthe mixtureto800C, *50C, ina water bath forapproximately30 minutesor untitlhetestarticldeissolves. 2. Once the testarticlheas dissolvedr,emove the suspensionfrom the water bathand spinthesuspensionwhileitcools.(Be surethereisa visible vortex,thiswillachievethe desiredemulsion.Thismay be preparedthe day beforeuse.) Writtenby:. Approved by- Date: 23-cr-;--')6 Clarification-:/No Y s (See attachedclarificatfioornm.) Initial/Date 418-014:PAGE C-'-"-2 ATTACHMENT 4 CROSS-FOSTERING PROCEDURE 418-014:PAGE C-37 ATTACHMENT 4 CROSS-FOSTERING PROCEDURE Protocol418-014 Page 1 of 1 The cohabitatiopneriodwilbleginone day earliefrortheratsassignedtothecontrol group. Dams assignedtothe controlgroup thatmate on the firsdtay willbe allowedto deliverand willretaintheirnaturalpups untinleeded forcross-fostednpgurposes. This willprovidea poolofcontroldams availablfeorimmediate cross-fostedntgopups from PFOS-treateddams, thus preventingthesepups from nursingfrom theirPFOS-treated dams. On the remainingdays ofcohabitationb,oth controland PFOS4reated groups willbe co-housed withmale breeders,one male rattoone female rat.Startinwgiththissetof deliveriesa,llpups willbe removed immediately(as soon as parturitiiosncomplete) from theirrespectivdeams and placedwitheitheranotherdam assignedtothe control group ora PFOS-treateddam. Ifa dam assignedtothe controlgroup isnotavailable, then a dam from the controlpoolwillbe used. The liftferom thatcontrolpooldam will then be sacrificevdiadecapitation. The firstten pups from the controlpool(irrespectiovfeliftedre)terminedtobe atday 1 postpartumwillbe decapitateda,nd the lungs(savedinBouin'ssolutiona)nd the liver (saved inneutralbuffered10% formalinw)illbe collecteadnd preservedforpossible futurehistopathologiceavlaluationT.he remainingpups from the controlpooldams will be sacrificevdiadecapitatioannd discardedwithoutnecropsyevaluation. Thiscross-fosteripnrgocedurewillresultinfourgroups of 12 dams/pups, i.e., Control/Contr(oSlubsetA),Control/PFOS(SubsetB),PFOS/Control(SubsetC) and PFOSIPFOS (SubsetD). Thisprocedurewillallowdistinctiotnosbe made between prenataland postnataleffectsofthecontinuousmaternalPFOS treatment. 418-014:PAGE C-38 ioPiP-LMF-,DICA Argus Reseamh LaboratoriesI,nc. 905 Sheehy Drive,BtildingA Horsham, PA 19044 Telephone:(215)443-8710 Telefax:(215)443-8587 PROTOCOL 418-014 Oral(Gavage) Cross-FosterinSgtudyofPFOS inRats SPONSOR'S STUDY NUMBER: T-6295.13 Amendment 1 - 4 January 1999 Cross-FosteringProcedure (Paragraph3 ofAttachment4 tothe protocol): The firsttenpups fromthecontrolpool(irrespectiovfelittearn)d thefirstten pups thatarefound dead (noautolysipsresent)perdosage group determinedto be atday 1 postpartumwillbe decapitatedatapproximately1 to 3 hours after birtha,nd the lungsand the livewrillbe retained.Several(minimum oftwo) sections(approximately1 mm thick)ofthelungand the livewrillbe removed usinga scalpeland willbe retainedinMcDowell-Trump'sFixative[preparation procedureprovidedby theSponsor isdocumented inthe raw data (storedunder refrigeratiionnscintillatviioanls()seeattached)f]orfutureevaluation.The remaininglungand liversamples willbe retainedinBouin'ssolutionor neutral buffered10% formalinr,espectivelfyo,rpossiblefutureevaluation.The remainingpups from the controlgroup dams willbe sacrificevdiadecapitation and discardedwithoutnecropsyevaluation. The livesrectionsinMcDowell-Trump'sFixativweillbe shipped(on coldpacks), forevaluationby electronmicroscopy,to: Jeanne deward PathologyAssociatesInternational 4915 D ProspectusDrive Durham, NorthCarolina27713 Telephone: (919)544-5257 Telefax: (919)544-3218 The remainingsamples willbe shipped(ambientconditionst)othe KrisJ.Hansen, Ph.D.,atthe address citedinthe protocolforhistopathological and biochemicalevaluation. 418-014:PAGE C-39 Reason forChange: Protocol418-014 Amendment 1 Page 2 The Sponsor has requested that these additionalsamples be retained for continued evaluation of the reasons forreduced pup survivalobserved in previous studies. 2. Cross-Fostering Procedure (Paragraph 4 of Attachment 4 to the protocol): The fourthparagraph of the Cross-Fostering procedure has been changed to read: This cross-fosteringprocedure willresultinfour groups of 12 dams/pups, i.e.,Control/Control(Subset B), Control/PFOS (Subset A), PFOS/Control (Subset D) and PFOS/PFOS (Subset C). This Procedure willallow distinctions to be made between prenatal and postnatal effectof the continuous maternal PFOS treatment. Reason forChange: This correctsthe protocol. a4l 0 Alan M. Hoberman, Ph.D., DABT Date Directorof Research "nd G. York, DABT Date hV-' Associate Director esearch and Study Director Dena C. Lebo, V.M.D. Date Chairperson, InstitutionaAlnimal Care and Use Committee Marvin T. Case, D.V.M., Ph.D. Study Monitor Date 418-014:PAGE C40 FIXATIVE PREPARATION Protocol418-014 Version:418-014 (29 Dec 98) Page 1 of 2 PROCEDURE Fixative McDowell-Trump's Fixative A. Purpose: The purpose ofthisprocedureistoprovidea method forthe preparation ofMcDowell-Trump's Fixativfeortissuesamples forstudy418-014. B. General Information: 1. Allsolutioncontainerswillbe labeled.Each labelwillspecifythe protocolnumber, fixativiedentificatiporne,parationdate,expiration date and storageconditions. 2. Fixativewillbe prepared: - Daily Weekly x Once 3. Safety: x Gloves,labcoat,gogglesorsafetyglassesand face shield x Dust-MistRespirator x Half-FaceRespirator Full-FaceRespirator/PositiPvreessureHood Tyvek Suit/Apron X Other:Chemical Fume Hood 4. Dosage suspensionsadjustedfor% Activity/PuriotryCorrection Factor: - Yes X _ No (Calculatiobnassed on 100%) 5. Sampling requirements:Citedinprotocol. 7. Storage:Room temperature 4IM14:PAGE C-41 FIXATIVE PREPARATION PROCEDURE Protoco4l18-014 Version:418-014 (29 Dec 98) Page 2 of 2 C. Preparationof Fixative: McDowell-Trump's Fixative 1 Add 860 mLs of R.O. Deionizedwater intoan appropriately labeledcontainer. 2. Add 100 mLs of37% Formaldehyde tothe containerand stiruntil uniform. 3. Add 40 mLs ofGluteraldehydetothe containerand stiruntil uniform. 4. Add 11.6g of Sodium Phosphate Monobasic to the container and stiruntil uniform. 5. Add 2.70g of Sodium Hydroxide to the vesseland stiruntiluniform. [NOTE: The finalsolutionshould have a pH of approximately7.3.) Written By: Approvebd42--l-@ Date: Clarification:,,/No _Yes [seeattachedclarificatifoonrm] Initials/Date: 2e, APPENDIX D DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY TEMPERATURE APPENDIX E AND RELATIVE HUMIDITY REPORTS 418-014:PAGE E-1 ARGUS Temperature and RelativeHumidity Report Location:Room 04 ProtocolNumber: 41"14 Range of Dates: 27-Oct-1998 13:15 to 23-Nov-1998 15:49 Target Range: Species: Rat Total Number of Days: Total Number of Houm: Total Number of Data Points: Temperature 64OF to 79OF 28 650.51 634 RelativeHumidity 30% to70% 28 650.51 634 Mean (tSD): Maximum: Median: Minimum: Number of Points in Range Number of Points High (%): Number of PointsLow (%): 69.1 76.4 68.4 66.5 634 0 0 1.9) (100.0) (0.0) (0.0) 53.2 66.2 57.2 28.6 633 0 1 8.0) (99.8) (0.0) (0.2) Report Generated: 04-Feb-1999 at 17:26 COMMENTS: REVIEWED BY: DATE: @-091 Cumulative by Location(v04.01.97) 418-014:PAGE E-2 ARGUS Temperature and RelativeHumidity Report Location:Room 27 ProtocolNumber: 418-014 Range of Dates: 23-Nov-1998 15:49to 31-Dec-I998 14:10 Target Range: Species:Rat TotalNumber of Days: TotalNumber of Hours: TotalNumber ofData Points: Temperature 64*F to 79*F 39 910.01 913 RelativeHumidity 30% to70% 39 910.01 913 Mean (SD): Maximum: Median: Minimum: Number of PointsinRange (%): Number of PointsHigh (%): Number of PointsLow 70.9 (:0t.7) 57.1 (:6t.1) 72.6 69.2 71.0 58.3 69.0 36.0 913 (100.0) 913 (100.0) 0 (0-0) 0 (0-0) 0 (0-0) 0 (0-0) Report Generated:04-Feb-1999 at17:28 COMMENTS: REVIEWED BY: DATE: Cumulativeby Lor-atio(nvD4.01.97) 418-014:PAGE E-3 ARGUS Temperature and RelativeHumidity Report Location:Room 35-37 Protocol Number: 418-014 Range of Dates: 31-Dec-I998 14:10 to 18-Jan-I999 08:45 TargetRange: Species:Rat TotalNumber ofDays: TotalNumber ofHours: TotalNumber ofData Points: Temperature 64*F to79OF 19 426.24 427 RelativeHumidity 30% to70% 19 426.24 427 Mean (SD): Maximum: Median: Minimum: Number of PointsinRange Number ofPointsHigh (%): Number ofPointsLow (%): 75.6 (1.7) 50.1 7.4) 78.3 56.1 76.4 51.6 68.5 5.0 427 (100.0) 417 (97.7) 0 (0-0) 0 (0-0) 0 (0-0) 10 (2.3) Report Generated:04-Feb-1999 at17:30 COMMENTS: REVIEWED BY: DATE: Cumulativeby Location(v04.01.97) 418-014:PAGE E-4 ARGUS Temperature and RelativeHumidity Report Location:Room 27 Protocol Number: 418-014 Range of Dates: 18-Jan-I999 08:45 to 29-Jan-I999 09:57 TargetRange: Species:Rat TotalNumber ofDays: TotalNumber ofHours: TotalNumber ofDataPoints: Temperature 64OF to79OF 12 265.0 266 RelativeHumidity 30% to70% 12 265.0 266 Mean (SD): Maximum: Median: Minimum: Number ofPointsinRange Number ofPointsHigh Number ofPointsLow 70.4 0.5) 54.3 72.0 70.4 70.4 56.4 69.1 33.1 266 (100.0) 265 0 (0.0) 1 0 (0.0) 0 8.3) (99.6) (0.4) (0-0) Report Generated:04-Feb-1999at17:33 COMMENTS: REVIEWED BY: DATE:. Cumulativeby Location(v04.01.97) 418-014:PAGE E-5 ARGUS RelativeHumidity DeviationsReport Location:Room 04 Protocol Number: 418-014 Range of Dates: 27-Oct-1998 13:15 to 23-Nov-1998 15:49 Humidity Target Range: Species: rat Date 03-Nov-1998 Time 14:00 R.H. 28.6 L 30% to 70% Date Time R.H. H = Value out of range -High L = Value out of range -Low R.H. = RelativeHumidity Report Generated: 13-May-1999 at10:45 L--,"These deviationsdid notadverselyaffectthe outcome orinterpretationfthe study. The followingdeviation(si)mpacted on the outcome of thestudy as described: Study Director Date: @9 t cj DeviatbiyoLnoscat(ivo0n4.01.97) 418-014:PAGE E-6 ARGUS RelativeHumidity DeviationsReport Location:Room 35-37 ProtocolNumber: 418-014 Range of Dates: 31 -Dec-I998 14:10 to 18-Jan-I999 08:45 Humidity Target Range: Species: rat Date 05-Jan-I999 05-Jan-1999 05-Jan-1999 06-Jan-1999 06-Jan-I999 06-Jan-I999 06-Jan-1999 06-Jan-1999 06-Jan-1999 06-Jan-1999 rime 21:00 22:00 23:00 00:00 01:00 02:00 03:00 04:00 05:00 06:00 R.H. 5.8 L 5.1L 5.0 L 5.1 L 5.1 L 5.2 L 5.3 L 5.5 L 5.6 L 5.6 L 30% to 70% Date Time R.H. H Value out of range - High L Value out of range - Low R.H.= RelativeHumidity Report Generated: 13-May-1999 at10:50 These deviationsdid not adverselyaffectthe outcome or interpretatiofnthe study. The followingdeviation(si)mpacted on the outcome ofthestudy as described: Study Director: Date: 13-Md@"/Qo) Deviationsby Location(v04.01.97) 418-014:PAGE E-7 ARGUS RelativeHumidity DeviationsReport Location:Room 27 Protocol Number: 418-014 Range of Dates: 18-Jan-1999 08:45 to 29-Jan-1999 09:57 Humidity Target Range: Species: rat Date 24-Jan-1999 Time 06:00 R.H. 70.4 H 30% to 70% Date Time RH. H = Value out of range -High L = Value out of range -Low R.H. = RelativeHumidity Report Generated: 13-May-1999 at 10:54 VI These deviationsdidnot adverselyaffectthe outcome or interpretationfthe study. The followingdeviation(si)mpacted on the outcome ofthe study as described: Study Director ots, .ks",4. u Date: -96 Deviationsby Location(v04.01.97) 418-014:PAGE D-1 DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 1 . Two extrafemale from thetreatedgroup (Group 11)were used for pharmacokineticsample collectioonn day 14 of lactation (DL 14),22 January 1999. These two female ratsinturnreceivedall collectionasnd were sacrificeodn DL 14. This deviationdid notadversely affectthe outcome ofthe studybecause the pharmacokineticsamples were not analyzed. 2. Eightfemale ratsratherthan sixfemale ratsfrom the controlgroup (Group 1) were used forpharmacokineticsample collectioonn DL 14,17 January 1999, 21 January 1999 and 22 January 1999. The two extrafemale ratsreceived allsample collectionasnd were sacrificeodn DL 14. Thisdeviationdid not adverselyaffectthe outcome ofthe study because the pharmacokinetic samples were not analyzed. 3. A totalof25 lifterfsrom the control(Group 1)and treated(Group 11)groups were cross-fostererdatherthan 24 liftersT.his deviationdidnot adversely affectthe outcome ofthe studybecause only one additionaliftewras added toeach dosage group. 4. Twenty-fivefemale ratsratherthan allofthe ratsinthe controlgroup (Group 1)were placed intocohabitatioonn 13 December 1998. The cohabitationperiodforthese ratslastedsixdays ratherthan fivedays.This deviationdid not adverselyaffectthe outcome ofthe studybecause it providedsufficienmtated ratsforcross-fostering. Alldeviationsare documented inthe raw data. . 'ACI-11 - -----2-,3 R .York, D., BT Date AssociateDirectorof Rese rch and Study Director APPENDIX F PATHOLOGY REPORT 418-014:PAGE F-1 Pathology Associates intemational A Company of Science ApplicationsInternationalcorporation An PATHOLOGY REPORT (ANCELLARY STUDY) ELECTRON NUCROSCOPIC EVALUANON OF LRVER AND LUNG IN CRL:CDOBR VAF/PLUSO RATS) ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS CLIM STUDY NUM[BER 418-014 PAI STUDY NUM[BER EM 99.46 Prepared by: James B. Nold, D.V.M., Ph.D.,Diplomate, A.C.V.P. Pathology AssociatesInternational 4915-D Prospectus Drive Durham, NC 27713 April5,1999 PREPARED FOR 3M CORPORATE TON3COLOGY, 3M CENTER, ST. PAUL, MN 55144-1000 4915 D ProspectusDrive9 Durham, North Carolina27713 9 (919)544-5257 o (919)544-3218 FAX .418-014:PAGE F-2 PathologyNarrative TransmissionElectronNEcrograph InterpretatiFoonrms TransmissionElectronNEcrographs QualityAssurance Statement Section I 11 m IV 418-014:PAGE F-3 1. PathologyNarrative 418-014:PAGE F-4 PATHOLOGY (ANCELLARY REPORT STUDY) AncillaryPathology Report Primedica Argus Study No. 418-014 ELECTRON NUCROSCOPIC EVALUATION OF LIVER AND LUNG IN CRL:CDOBR VAF/PLUS@ RATS) ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS CLIENT STUDY NUMEBER 418-014 PAI STUDY NUMEBER EM 99.46 SUNEM9@URY Ultrastructuraltlhye,rewere increasednumbers of peroxisomeswithinhepatocytesof neonatalratpup liversfrom dams treatedwith 1.6mg/kg/day (bLigh-doseP)FOS compared with controls.Quantitatioonfperoxisomesindicatedslightlmyore thantwice thenumber of peroxisomesin treateadnimals.Differencesin cellulamrembranes or mitochondriawere notdiscerniblbeetween controland treatedanimalsin thesamples examined. Type 11pneumocytes containinglamellarbodieswere identifieidnboth controlsand high-doseanimals;however, thereappearedto be an increasednumber of type H pneumocytes and lamellarbodiesin lungsfrom treatesdamples. Although some lamellarmaterial(surfactanwta)s identifiabwliethinalveolarlurr@nan,o significant differencewas notedbetween controland treatedsamples. PROCEDURES The objectiveof thisstudywas toexamine by electronmicroscopytheliversand lungs from selectedday-I postpartumratpups inan oral(gavage)cross-fosterisntgudyof PFOS. ln additionto a generalqualitativeevaluationfordifferencebsetween controland treatedanimals,theultrastructureanldpointsincludedquantitatioonf peroxisomeswithin hepatocytes,membrane and mitochondrialchanges inhepatocytesa,nd the quantitatioonf surfactanltamellarbodiesinthelung. For thecross-fosterisntgudy,66 femaleCrl:CDOBR VAF/PLUSO ratswere giventhe testarticloer thevehicleonce dailybeginning42 days priorto cohabitatiownith a male throughday 21 postpartumaccordingtoText Table 1. Text Table 1. Dosage Levels,Concentrationsand Volumes Dosage Group Number of Mated Female Rats 30 + 6' PFOS Dosage (mg/kg/day) 0 (Vehicle) PFOS Concentration (MR/ML) 0 Dosage Volume (MLAG) 5 24 + 6a 1.6 0.32 5 a Samples foranalysisof pharmacokinctics were collectedfrom 6 dams and theirrespectivelitterosn day 14 postpartum. These dams and theirrespectivelitterwsere sacrificefdollowingsample collection. 1-1 418-014:PAGE F-E AncillaryPathology Report Primedica Argus Study No. 418-014 The firsttenpups from thecontrolpool(irrespectiovfelittearn)d thefirst10 pups that were founddead (no autolysipsresentf)rom thetreatedgroup determinedtobe atday I postpartumwere decapitateadtapproximately1-3hoursafterbirth,and thelungsand liverwere collected.Samples of lungand liverwere thin-sliceadnd placedinMcDoweUTrump fixativ(eMcDowell EM, 1976),and submittedtothislaboratorfyorelectron microscopy processingand evaluation. Per sponsor request,tissuesamples from selectedpups were processedand evaluated ultrastructura(lTleyxtTable 2).Pup numbers were assignedby thislaboratorybecause submittedsamples were identifiebdy dam number only.The fivedigitnumber isthe dam number; thesuffixI or 2 denotesthepup number from thatdam's litter. Text Table 2. Animals SelectedforElectronMicroscopy Control(0 mg/kg/day) 18904-1 18904-2 18915-1 18915-2 18912-1 High-Dose (1.6mg/kg/day) 18952-1 18952-2 18956-1 18973-1 The lungsand liverswere thenprocessedintoSpuff'sresinforultrastructural examination.Thick (1g)sectionswere cut,stainedwithtoluidinbelue,and examined to selectrepresentativaereasforfurtherelectronmicroscopyprocessing.Thin sections (approximately90 nm) were cut,mounted on 200-mesh coppergridss,tainedwith 5% methanolicuranylacetateand Reynold'sleadcitratea,nd examined on a Zeiss900 transmissioenlectronmicroscope.Centrilobulahrepatocytesw,here clearlyidentifiable inliversectionsw,ere preferentiayexamined. Representativeelectron photorr@crographsof liverand lungwere takenand significanutltrastructurfaelatures were summarized foreach photographand animalon a designatedtransn-Lissieolnectron micrograph interpretatifoonrm. The number of peroxisomesinhepatocyteswas manually counted foreach centerphotographedhepatocyteand recorded. RESULTS AND DISCUSSION Individualinterpretatioonfslightand electronmicrographsforeach animal are shown in Section11.Selectedrepresentativeelectronphotomicrographswere labeledand are shown inSectionM. Fixationof thetissueswas lessthanoptimalforelectronmicroscopy.This was especially evidentin thelivercompared withthelung.Lung isinberentllyessdense thanliverand allowsbetterinfiltratioofnthefixativientothetissue.Suboptimalfixatiownas characterizebdy discontinuousplasma membranes, lossof detailof cytoplasmic organellesd,ilatatioonf endoplasmicreticulumand thenuclearenvelope,and dilatation of niitochondriaclristae.Fixationh,owever,was stilclonsideredadequateto assessfor 1-2 418-014:PAGE F-6 AncillaryPathology Report Primedica Argus Study No. 418-014 thepresenceofperoxisomesinthehepatocytesand lamellarbodiesinthelungand to make generalcomparisonsbetweencontroland treatedsamples. LRVER Control (0 mg/kgtday). Subcellular features of control hepatocytes are demonstrated in Figures I and 2. Hepatocytescharacteristicalcloyntainnumerous mitochondriaand prominent endoplasmic reticulum.In thesecellsrough endoplasmic reticulumwas evident,but smooth endoplasmic reticulumwas not because of suboptimalfixation. Nlitochondriagenerallyappeared dark and contained scattereddense depositsand some dilatedcristaeb,oth artifactdsue tosuboptimalfixation.Dilatationof endoplasniic reticulumand thenuclearenvelope were additionalartifactosf poor fixation.Fine granularmaterialinthe cytoplasm isconsistentwith glycogen particlesP.eroxisomes were identifieadnd quantitatedinfrom representativheepatocytes.For thecontrollivers, the average number of peroxisomes counted per hepatocytephotograph was 7 peroxisomes per cell(TextTable 3). Fetaland neonatalratliverisa hematopoietic tissue,and numerous hematopoieticcellswere evidentwithin theliversinusoids. Text Table 3. Quantitationof HepatocellularPeroxisomes Control(0 mg/kg/day) Number of Peroxisomes Animal Number per Hepatocvte 18904-1 4.6 18904-2 6.2 18912-1 9.0 18915-1 8.8 18915-2 6.4 L -Mean Control 7.0 Hil2h-D se (1.6mg&g/day) Animal Number 18952-1 18952-2 18956-1 18973-1 Number of Peroxisomes per Hepatocvte 16.0 17.0 17.25 14.0 1 1 1 Mean High@Dose = 16. High-Dose (1.6mg/kg/day). The averagenumber of peroxisomesper hepatocyte(14.7) was higherin theliigh-dosaenimalscompared withcontrols(Text Table 3). Otherwise, generalultrastructurmaolrphology was similarin high-doseliverscompared with control livers.Figures3-5 illustrartepresentativheepatocytesfrom severalhigh-doseanimals. Differencesin mitochondriaand cellulamrembranes were not discerniblbeetween highdose and controllivers.However, suboptimalfixationof thetissueprecludeddetailed evaluationof theseorganelles. LLTNG Control (0 mglkg/day). Lung samples were atelectatiacnd not inflatedwith fixative. Thereforemost alveoliand alveolarseptawere collapsedwith poor delineationof alveolarand airway spaces. Figures6-8 illustratseome normal cellularitaynd ultrastructurpaullmonary anatomy. Type 11pneumocytes (PN2) were infrequently identifieldiningalveolit,erminalbronchioleso,r withincollapsedsepta.Type 11 pneumocytes.were identifiedby thepresence of lamellarbodies,which arecytoplasmic 1-3 418-014:PAGE F-7 AncillaryPathology Report PrimedicaArgus Study No. 418-014 vacuolescontainingelectron-densleaminatedmembrane-likeprofilesL.ainellarbodies representthe materialsecretedintothealveolarspacesas surfactantS.urfactantor lamellarmaterialwithinairwayspaceswas very rarelyidentifieidncontrollungs.The typeIpneumocyte (PN 1)isch-grateristicalflyatteneadnd linesthealveolarspace as a thinmargin of cytoplasm.Type Ipneumocytes differentiaftreom typeIIpneumocytes. Many pneumocytes appearedto be cuboidalwithoutdistinctivleamellarbodies,and may representa transitionstage between PN2s and PN Is,especiallysincethese lungs were neonatal and had not been fullyexpanded with air.Additionally,some cuboidal epithelial cellsrepresentedterminal bronchiolarepithelium. Type H pneumocytes and larneuarbodies were infrequentlyidentifiedin these control lungs and no reliablequantitationcould be assessedof them. Some lamellar-likebodies were also presentwithin capillarylumina. lEgh-Dose (1.6mgtkgtday). Ulautructural pathology of the high-dose lungs was essentiallysimilarto controlsamples. There appeared, however, to be increased numbers of type IIpneumocytes and lameuar bodies;atleast,identifyingthem forillustrative purposes was easierthan in controlsamples. The collapsednatureof most of the alveoli and septa made definitivequantitationimpractical.Some lameuar bodies were identified within capillarylurainaand only a small number were seen within alveoli.Figures 9-12 illustratseome representativelung fieldsfrom the high-dose animals. CONCLUSION Ultrastructuraelvaluationof liverand lungs of ratpups from dams treatedwith 0 (control)or 1.6 mg/kg/day (higb-dose)PFOS identifieidncreasednumbers of peroxisomes within hepatocytes of high-dose ratpup livers.Quantitationof peroxisomes indicatedslightlymore than twice the number of peroxisomes in treatedanimals. Differences in cellularmembranes or mitochondria were not discerniblebetween control and treatedanimals in the samples examined. In lung samples the presence of type II pneumocytes and lameuar bodies was evaluated.Type IIpneumocytes containing lamellarbodies were identifiedin both controlsand high-dose animals; however, there appeared to be an increased number of type H pneumocytes and lamellarbodies in lungs from treatedsamples. Some lamellarbodies and lamellarmyelin-likefigureswere extracellulaarnd within capillarylumina. Although some lamellarmaterial(surfactant) was identifiablweithin alveolarlumina, no significandtifferencewas noted between control and treatedsamples. SIGNATURE OF AUTHOR 9, n 49PA '4ames B. Nold, D.V.M., Ph.D., Diplomate, A.C.V.P. @x Date' 1-4 418-014:PAGE 1'7-8 AncillaryPathologyReport PrimeciicAargus Study No. 418-014 REFERENCES McDowell, EM and Trump, BF: H,@stologifcixativfeorroutinediagnosticlightand electronmicroscopy.Arch.PathoL Lab.Med, 100:405-414, 1976. -.1'8t--014:PAGE F-9 11.TransmissionElectronNficmgraphInterpretatiFoonrms' 1Fon-nsaretypographicallcyon-ectedversionsofsigned/daterdaw datasheetsmaintained inarchivedstudyfile. 418-014: PAGE F-1C TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-014:EM-99.46 Animal No.: 18904-1 TreatmentGroup: -0 mg/kg Control Sex: non arr)licable Species/StraiCnr:l:CDSBR VAF/Pluse Rat Tissue-:Liver BlockNo(s).: 99.46-1 Z15683,Zl5684, Photo/NegativNeo(s).:Z15686-88 SignificLaenstion(schecokne):_ Yes X No InterpretiPnagthologis(tsignaturaend date): Features: Z15683:-Hepatocytes,everalerythrocyteisnsinusoida,nd nucleusof probableendotheliaclellon rightsideofphotograph.Six(6)peroxisomesevidentin largecentrahlepatocyte.DilatedblebsinRER arepseudo-peroxisomes.Abundant fine granularmaterialg,lycogen,incytosol.Dense mitochondriawithsome granulardeposits and dilatecdristaeP.oor definitioonfendotheliac.e1lls. Z 15684: Severalhepatocytincucleipresentb,utpoorcytoplasmicmargindefinition. Some swellingof n-LitochondrDiial.atedblebsinRER. Bilecanaliculuisnlowerleft comer. Two (2)peroxisomesincentraclell.Inadjacentwo cellsonlysingle peroxisomesareclearlyevident. Z 15686: Singlehepatocytesurroundedby erythrocytesN.o cleardefinitionf endotheliaclells.Distendedcytosolwithglycogen-likgeranularparticlesA.bundant mitochondriand RER. Mitochondriahave some granulardensitieasnd dilatecdristae. DilatedblebsinRER. Ten (10)peroxisomesevident. Z15687: Singlehepatocyte.GranulocyticceU (hematopoietipcr)esentinbottomleft comer. DistendedcytosolwithgranularmaterialD.ense mitochondriwaithsome granulardepositsT.wo (2)peroxisomesevidentincentrahlepatocyte. Z15688: Hepatocyte.ErythroidprecursorpresentinlowerleftsinusoidD.ilatedblebsin RER. Cytosolexpandedwithfinegranularmaterialp,robablyglycogen.Three(3) peroxisomespresent. Conclusions:Normal hepatocytes2.3/5= 4.6peroxisomesperhepatocytevident. Plasma membranes poorlydefined.Dense rr@tochondriwaithsome granulardepositsand dilatecdristaeS.uboptimalfixation. 418-014:PAGE F-11 TRANSNUSSION ELECTRON NUCROGRAPH ENTRPRETATION StudyNo.: -418-014:EM-99.46 Animal No.: 18904-1 TreatmentGroup: 0 mg/kg Control Sex: non at)t)licable Species/StrainC:rl:CDSBR VAFfPlusM) Rat Tissue: Lung BlockNo(s).: 99.46-2 Z15689 - Photo/NegativNeo(s).:Z15693 SignificanLtesions(checkone): Yes x No InterpretiPnagthologis(tsignaturaend date): Features:Z15689: Capillarwyithdme endotheliacleU nuclei.Inlumen ofcapillariys a lamellar-libkoedy. SeveraltypeI pneumocytes(PN I)presentsurroundingcapillary. No pneumocyte lamellarbodiespresent. Z15690: Type 2 pneumocyte(PN2)presentc,ontaininmgultiplleamellabrodies(-7). BeneaththePN2 isa capillarcyontaininagn erythrocytep,lateleatn,d endotheliaclell nuclei.PN I torightsideofphotographwithmore translucenctytoplasm. Z 15691: Collapsedalveolarsepta.On topisa PN 1 withflattenecdytoplasmand large round nucleus.Otherceuslessclearldyefined,butappeartobe primarilyPN Isin collapsedsepta.No pneumocyte lamellarbodiesevident. Z15692: Collapsedalveolasrepta.On topisathin-layerePdN1, and inthebottom left islargerprofiloefa PN 1. Centrallayppearstobe a capillarlyumen containingsome lamellardebrisand two leucocytesA.lsosome lipidebrisispresentinthecapillary lumen.Endotheliaclellcytoplasmisevidentb,utno endotheliaclellnucleus.No pneumocyte lamellarbodiesidentified. Z15693: Collapsedalveolasrepta.At topisa profiloefa PNl inthealveolasrpace. Beneathitisathin-layerePdNI, andtotheleftsideofphotoisa PN1. Subjacentnuclei areprobablyendotheliaclellnuclei.At bottomlefitsan erythrocyteJ.usttotherightisa collapsedalveolaind thin-linPeN 1.No pneumocyte lamcuarbodiesidentified. Conclusions: Normal lungtissueF.ixationadequate.Only one PN2 identified. 418-014:PAGE F-12 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-014:EM-99.46 Animal No.: 18904-2 TreatmentGroup: 0 mvAg/ Control Sex: non at)vlicable Species/StraiCnr:l:CIDSBRVAF/Pluse) Rat Tissue: Liver BlockNo(s).: 99.46-3 Z15694 - Photo/NegativNeo(s).:Z15698 SignificanLtesions(checkone): Yes x No InterpretiPnagthologis(tsignaturaend date): Features: Z15694: Hepatocyte.Multiplemitochondrias,ome RER and multiple roundedblebsofRER. Cytoplasmfullofgranularglycogen.PoorendotheliacleR definitioinnsinusoidsN.ucleatederythrocytienbottom leftM.itochondriaappeardense withsome granuladrepositsanddilatecdristaeT.hree(3)peroxisomesidentified. Z15695: Hepatocyte.DilatedRER. Multipledensen-,Litochondwriitahgranular depositsand dilatecdristaeC.ytoplasmicmarginsand endotheliaclellspoorlydefined. Six(6)peroxisomesidentified. Z15696: Hepatocyte.Many dilatebdlcbsofRER. Dense rnitochondrwiiathgranular depositsand dilatecdristaeT.op lefthepatocytehasswollen,lessdense,mitochondria. Twelve (12)peroxisomesidentified. Z15697: Severalpoorly-fixehdepatocytesM.any lightand darkdilatebdlebsofRER, which aredifficutlotdifferentiaftreom peroxisomes.Five(5)peroxisomesidentifieidn singlemore-circumscribehdepatocyte. Z15698: Hepatocyte.Largeclearareasincellcontainfinegranulamrateriaclonsistent withglycogen.Five(5)peroxisomesevident. Conclusions:Normal hepatocytesS.uboptimalfixation3.1/5= 6.2peroxisomesper cell. 418-014:PAGE F-13 TRANSMISSION ELECTRON MICROGRAPH DrrERPRETATION StudyNo.:-418-014,EM-99.46 Animal No.: 18904-2 TreatmentGroup: -0 mg/kg Control Sex:-non applicable Species/StraiCnr:l:CD(DBRVAF/PlusS Rat Tissue: Lung BlockNo(s).: 99.46-4 15699- Photo/NegativNeo(s).:-Z 15704 SigrlifiLceasniton(schecokne):_ Yes InterpretiPnagthologis(tsignaturaend date): x No Features: Z15699: Alveoluscontaininsgeveralcuboidalpneumocytecells.The lower cellappearstocontaina couplesmalllamellarbodies,probablya PN2. The othertwo largecuboidalcellsdo notcontainany lameuarbodies,butappeartobe pneumocytes which haven'tflatteneodutintoPNls orareterminablronchiolaerpitheliaA.t theleft and bottom arecapillarileisnedby endotheliaclells.The flattenePdN I atthebottom containsa segment ofluminalsurfacewithsome denseelectron-densmeaterialS.ome dilatecdristaienmitochondria. Z15700: Alveolarspacewithcapillary/erythroattylteeftandpneumocytesaroundtop and right.Top leftpneumocyte containsthree"empty"vacuolesp,ossiblyextruded lamellarbodies.PN I atrightisslightlmyore electrondense. Z15701: MultiplePNIs lininaglveolasrpaced.No lameuarbodiesorPN2s evident. Capillaryatlowerrightwithendotheliaclells. Z 15702: Celltypecontainingcentralarneuabrody notclearldyiscernibleI.tappearsto be outsidethecapillaraynd endotheliaclellattopleftb,utcompressedbetweenthe pneumocyte on rightand lowercentraclell.Thisisan areaofcollapsedalveolarsepta. Z15703: Alveolusand linincgeus.LargePNI appearstocontaina singlelamellarbody in centerof photograph.Capillaraynd endotheliumatleftof photograph.Collapsed alveolarseptainbottom centerand right.Dilatedcristaiensome n-iitochondria. Z15704: Alveoluswithlininpgneumocytes.CeU inlowerleftcomer containspartiallyempty vacuolecontainingsome lamellarmaterialD.ensercellatcenterleftcontainsa coupleclearvacuoles,althoughthiscellislinedby a flattenePdN 1. Conclusions:Normal lungand alveoli.A few lainellabrodiesidentified. @,4 -81-1014: PAG E F-14 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-014:EM-99.46 Animal No.: 18912-1 TreatmentGroup:-0 mg/kg.Control Sex: non applicable Species/StraiCnr:l:CDMBR VAF/Plusa Rat Tissue: Liver BlockNo(s).: 99.46-5 Z15773 - Photo/NegativNeo(s).:Z 15777 SignificLaenstion(scheckone):_ Yes X No InterpretiPnagthologis(tsignaturaend date): Features:Z15773: Hepatocyte.Some segmentsofhematopoieticcellsarepresentin theupperrightcomer. Endotheliaclelldetailasreindistincatn,dfixatioinspoor.There isdilatioannd blebbingoftheRER and some dilatioonfthenuclearenvelope.Six(6) peroxisomescounted. Z15774: Hepatocytesurroundedby some hematopoieticcellsM.arked dilatioannd blebbingof RER. Four (4)peroxisomescounted. Z15775: Hepatocytewithcanaliculuastthebottomand sinusoidaelrythrocytaettop right.Endotheliacleuspoorly-defineadnd fixatioinssuboptimalE.leven(11) peroxisomescounted. Z15776: Hepatc>cytiespoorly-fixeadnddisruptedM.arked dilatioanndblebbingofthe RER. Nfitochondriaaredenseand containsome granulardepositsand dilatedcristae. Seven (7)peroxisomescounted. Z15777: Hepatocyteappearssimilatrothosedescribedabove. Itispoorlyfixedand RER and nuclearenvelopearedilatedS.eventeen(17)peroxisomescounted. Conclusions:SuboptimalfixationN.ine (9)peroxisomes/hepatocyt(eaverageof5 cells) 418-014:PAGE F-'15 TRANSMISSION ELECTRON AACROGRAPH INTERPRETATION StudyNo.: 418-014:EM-99.46 Animal No.: 18912-1 TreatmentGroup:-0 mglkg.Control Sex: non aprlicable Species/StraiCnr:l:CDMBR VAF/PlusS Rat Tissue-:Lung BlockNo(s).: 99.46-6 Z15768 - Photo/NegativNeo(s).:Z15772 SignificLaenstion(scheckone):- Yes InterpretiPnagthologis(tsignaturaend date): X No Features:Z15768: A row ofcuboidalepithelicaelllsp,robablybronchiolaerpithelium. The centralsmauer cellcontainseveralempty vacuoleswhich may have been lamellar bodies,suggestiveofa PN2. Some distendedcristaienmitochondria. Z 15769: Again,severalcuboidalepitehliiclellsc,onsistenwtithbronchiolaerpithelium. The two centraclellscontaina few lam'ellarbodiesand areprobablyPN2s. The smaller darkercellappearstobe a basalcell. Z15770: Alveoluswithlinincgellsand underlyincgollapsedalveolasrepta.The small darkliningcellcontainsfourlamellabrodiesandisa PN2. The othertwo liningcellsare PNIS. Z15771: Area seems tobe a terminablronchiolejunctioniwnigthan alveolaarreawitha flattenePdNI (lowerright)C.entertopisa PN2 withfivelamellabrodies.Itappearsto squeezeinbetweentwo cuboidalepithelicaelllsA.t therightand leftarecapillarieTsh.e rightone containsa lamellarbody initslumen. Z15772: Two largecuboidalepithelicaelllsorpneumocytesand a PNl liningcell (bottom).Some dilatecdristaienniitochondrnioated. Conclusions: Appearsnormal.A few random PN2s withlamellabrodiesnoted. 418-014:PAGE F-16 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-014-E.M-99.46 Animal No.: 18915-1 TreatmentGroup: 0 mg/kg.Control Sex: non gpvlicable Species/StraiCnr:l:CDSBRVAFOus(S Rat Tissue: Liver BlockNo(s).: 99.46-7 Z15763 - Photo/NegatiNvoe(s).:-ZI5767 SignificLaenstion(schecokne):_ Yes X No InterpretiPnagthologis(tsignaturaend date): Features:Z15763: Hepatocyteinphotographhasa largeareaofcytoplasmfillewdith abundantglycogen-likmeaterialP.lasmamembranes and endotheliacleus arepoorlydefineddue topoor fixationN.Etochondriaaredenseand containsome densedeposits and dilatedcristaeT.here isdilatioonftheR.ER,showing asround blebson cross-section. A bilecanaliculuisspresentinlowerleftF.ourteen(14)peroxisomescounted. Z15764: Hepatocyte,similatroZ15763. Five(5)peroxisomescounted. Z15765: Hepatocyte.Poor fixatiocnlearleyvidencedby lossofendotheliaclellslining sinusoidsE.ight(8)peroxisomescounted. Z 15766: Six(6)peroxisomescounted.Much ofthecytoplasmcontainsfinedispersed granulesp,robablymostlyglycogen,butalsoconsistenwtithpoorfixation. Z15767: Hepatocytesin-Liltaorthosedescribedabove. Many dilatedblebsofRER. Eleven(11)peroxisomescounted. Conclusions:Suboptimalfixatioonftissue.Number ofperoxisomescounted: 8.8/cel(lfivecounted). 418-014:PAGE F-17 TRANSMISSION FLECTRON MICROGRAPH WMRPRETATION Study No.: -418-014: EM-99.46 Animal No.: --18915-1 Treatment Group: 0 mg/kg. Control Sex: non avrlicable Species/StrainC:rl:CDeBR VAF/Pluse Rat Tissue:- Lung Block No(s).:- 99.46-8 Z15758 - Photo/NegativeNo(s).:-15762 SignificanLtesions (checkone): yes InterpretinPgathologist(signatureand date): X No Features: Z15758: Collapsed alveolarsepta.Air spaceatbottom islinedby a thin electron-densecytoplasmicmargin containinga few organellesand a largedilatedbleb. Two capillarieisnthe collapsedareacontainsome lamellarstructuresfreewithin the lumina. Lamellar bodies withinpneumocytes or freeintheairspacesare not evident. Z15759: Alveolusand septa,mostly collapsed. PNIs lineairspaces.Capillaryattop leftappearstocontaina lameuar body withintheendothelialcellcytoplasm. Smaller dark nucleus inthe centerappearstobe a nucleus of a PN I. Z15760: Area of collapsedsepta,with airspaceto left.The spaceislinedby thinmargin of PN1. Subjacentto thisspace isa capillarycontaininga largelamellarbody within the lumen. At top rightisa cell,probablya PN2, which containsa singlelamellarbody inits cytoplasm. The clearceU in thecenterappears tobe a pneumocyte. Z 1576 1: Photograph containsseveralcapwaries in thecollapsedsepta.The largelumen atthebottom containssome pale-staininlgipidmaterialintermixedwith endothelialcell cytoplasm. Air space islinedby flattenePdN I cytoplasm.No lameuar bodies noted in pneumocytes. This profileinlumen may be tubularsurfactant(lamellarbody). Z15762: Centraldark cellappearsto be a PN2 with a few lameuar bodies in its cytoplasm. FlattenedPN I cytoplasm linestheremainderof theairspace.Two capillary lumina,dorsaltoand to therightof thisPN2, containsome lamellarmaterial.A focusof lamellarmaterialisinthe airway lumen. Conclusions: Normal fetallung.One PN2 was identifiewdith some lamellarbodies. Other lamellarlikestructureswere freewithincapillaryluniina,and a singlefocusof lamellarmaterialwas presentinairway lumen. 418-014:PAGE F-18 TRANSAUSSION ELECTRON AUCROGRAPH DMRPRETATION StudyNo.:-418-014:EM-99.46 Animal No.: 18915-2 TreatmentGroup:0 mg/kg.Control Sex: non aRRlicable Species/StrainC:rl:CM@OBRVAF/Plus(D Rat Tissue: Liver BlockNo(s).9:9.46-9 Z15753 - Photo/NegativNeo(s).:Z15757 SignificLaenstion(schecokne):_ Yes x No InterpretiPnagthologis(tsignaturaend date): Features:Z15753: Hepatocytewithnumerousaated blebsofRER present.Some dilatationostedinnuclearenvelope.Abundant granularglycogenincytoplasm. Nfitochondriaappeardensewithsome granulardensitieasnd some dilatedcristaeN.ine (9)peroxisomescounted. Z15754: Hepatocytesurroundedby erythrocyteasnd hematopoieticcellsE.ndothelial cellsarepoorlydefineddue topoorfixationH.epatocytecontainsmany dilatesdegments and blebsofRER and nuclearenvelope.Five(5)peroxisomescounted. Z15755: Largecentralhepatocytewithabundantglycogen.BlebbingofRF-R abundant. Two (2)peroxisomescounted. Z 15756: Fieldwithseveralhepatocytesh,owever,onlyone attopcenterwithnucleusisa completecell.Cellshas numerous dilatebdlebsofRER, some darkand some light stainingS.ome dilatecdristaneotedinrriitochondriFao.ur(4)pcroxisomescounted. The cellatlowerrightappearstohavemore pcroxisomes,butisan incompletecellfor counting. Z 15757: Severalhepatocytesa,lthoughcentercellwithnucleusisonlycompleteceU. It has densemitochondriawithgranulardensitieasnd numerous dilatebdlebsof RER. Twelve (12)peroxisomescounted. Conclusions:Averagenumber ofperoxisomes= 6.4(5ceuscounted).Suboptimal fixation. 418-014:PAGE F-19 TRANSAUSSION ELECTRON AUCROGRAPH DFMRPRETATION StudyNo.: 418-014:EM-99.46 AnimalNo.:- 18915-2 TreatmentGroup:-0 my Ag. control Sex: non Mlicable SPecics/StraiCnr:l:CDOBR VAF/Pl,,s(v Rat Tissue: Lung BlockNo(s).-: 99.46-10 Z15748 - Photo/NegativNeo(s).:-Z15752 SignificaLnetsions(checkone): Yes InterpretiPnagthologis(tsignaturaend date): x No Features:Z 15748: Two alveolwiithinterveninsgeptum andcapillaryP.N Is linethe septum,and thereisa largecellattachedtotheright.Thiscellhasminimalcytoplasmic organeues,mightbe a lymphocyte. Some lameuarmaterialispresentwithinthe capillarlyumen,butisnotpresentinpneumocytesin thisphotograph. Z15049: Alveolus,liningcellsa,nd collapsedsepta.Liningpneumocytecontainsa singlelamellarbody initscytoplasmjustbeneaththenucleus.At centratlopareseveral lamellarbodieswith cytoplasmof probablyPN2s. Z15750: CuboidalepitheliuomfprobablybronchioleC.entralliysa PN2 with5 lamellar bodies. Z15751: Alveolusand lininPgMls. At lowerrightisa capillarwyitha lamellabrody in thelumen ofthecapillaryA.t toplefti,ncollapsedseptaairea,isa PM2 withseveral larnellabrodies. Z15752: Alveoluson leftwithlininpgneumocytesisfreeoflamellabrodies.At topright isa singlecellwiththreevacuoleswhich appeartohavelainellasrtructureisnthem.Two capillariaetscenterand lowerright. Conclusions:SeveralPN2 and lamellarbodiesidentifiedS.ome lamellarbodieswere in capillarlyumina. 418-014:PAGE F-20 TRANSMISSION ELECTRON MICROGRAPH MMR,'.DRETATION StudyNo.:-418-014:EM-99.46 Animal No.:- 18952-1 TreatmentGroup: 1.6mg/kg. high-dose Sex: non applicable Species/StrainC:rl*CD(E)BVRAF/pluse Rat Tissue-:Liver BlockNo(s).: 99.46-15 Z15722 - Photo/NegativNeo(s).:Z15726 SignificanLtesions(checkone): X Yes No InterpretinPgathologis(tsignaturaend date): Features:Z 15722: Hepatocyte.Cytoplasmcontainsabundantfinegranularmaterial, glycogen.Some dilatedblebsof RER. NEtochondriaappeardense withgranulardeposits and some dilatedcristaeT.wenty (20)peroxisomesidentifiedA.djacentendotheliaclell ispoorly-define(dpoorfixation). Z15723: SimilartoZ15722. Numerous rounddilatebdlebsofRER. Bile canaliculus presentin bottom of photograph. Ten (10) peroxisomes counted. Z15724: SinusoidcontainingseveralerythrocytesS.egment of hematopoietic granulocyticcellintopleft.Endotheliaclelland lipid-containipnegrisinusoidaclellare poorlyfixed.Two incompletehepatocyteson right.A few peroxisomesarenoted,but not counted. Z15725: Hepatocyte.Bilecanaliculuasttop;sinusoidinlowerrightofphotograph. Numerous dilatedblebsof RER. Fifteen(15)peroxisomescounted. Z15726: Hepatocyte.Bilecanaliculaittop rightand left.Numerous, variably-sized dilatedblebsofRER. Nineteen(19)peroxisomescounted. Conclusions: Increasednumbers of peroxisomes. Average 16 peroxisomes/hepatocyte (4 cellscounted).Suboptimalirixation. 418-014:PAGE F-21 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.:-418-014:EM-99.46 Animal No.:- 18952-1 TreatmentGroup: 1.6mg/kg.high-dose Sex: non gprlicable Species/StraiCnr:l:CDO13RVAF/Pluse Rat Tissue: ung BlockNo(s).: 99.46-16 Z15717 - Photo/NegativNeo(s).:Z15721 SignificaLnetsions(checkone): x Yes No InterpretiPnagthologis(tsignaturaend date): Features: 15717: AlveoluswithliningceflsT.wo largePNIs arepresentand appearto have a PN2 betweenthem. However,a PN1 marginstilsleparatetshePN2 from the sinusoid.SeverallamellarbodiesarepresentassociatewdiththePN2, and some ofthem appeartobe extracellulaTrh.e PNI on theleftcontainsabundantfinegranularmaterial in itscytoplasm,consistenwtithglycogen. Z 15718: Collapsedareaof alveolasrepta.Two centraclellscontainlamellarbodies, three(3)inone,and seven(7)intheother.Both cellsappeartobe PN2s,although normal pulmonary architecturiesunclearbecauseofthecollapsedstate. Z15719: Two cellswithabundantcentrioleasppeartorepresenbtronchiolacrellsa,nd thecentrioleasreassociatewdithciliaC.entrallayretwo cuboidalcells.One contains multiple(5)lamellabrodies.Thisccilappearstobe a PN2. A largelamellabrody is associatewdiththeleftcellb,utitmay be extracellulaBro.th cellscontainabundant granularmaterialc,onsistenwtithglycogen. Z 15720: Alveolusand linincgells.Two cellscontainlamellabrodies,sixinone and eightintheother.Some lamellarmaterialappearstobe extracebular. Z15721: Alveolus,fivelinincgellsa,nda capillariyntoprightofphotograph.Thisis possiblyajunctionwitha terminablronchiole.Two ofthepneumocytescontainmultiple lamellarbodies,and appeartobe PN2s. Conclusions:There appeartobe increasendumbers ofPN2s containinglarnellabrodies. Some cxtracellullaarmellarmaterialnoted. 418-014:PAGE F-22 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: -418-014,EM-99.46 Animal No.:-18952-2 TreatmentGroup: 1.6my,/kg.tligh-dose Sex: non avrlicable Species/StrainC:RECDSBR VAF/PlusS Rat Tissue: Liver BlockNo(s).: 99.46-17 Z15712 - Photo/NegativeNo(s)-.Z:15716 SignificaLnetsions(checkone): x Yes No InterpretiPnagthologis(tsignaturaend date): Features:Z15712: Hepatocyte.Poorly-defmedsinusoidasnd endotheliumattopright and bottom ofphotograph.The hepatocytecytoplasmappearsvoluminouswithabundant finegranularmaterialconsistenwtith glycogen.Nutochondriaappeardense and.have some granulardepositsanddilatecdristaeS.ome dilatebdlebsofRER. Eight(8) peroxisomesidentified. Z15713: Hepatocyte.HematopoieticceU torightB.ilecanaliculupsresenitnlower right.Abundant finegranularmaterialg,lycogen,incytoplasm.Appear tobe increased numbers of peroxisomes.Twenty-seven(27)peroxisomespresent. Z15714: Hepatocyte.Many dilatebdlebsofR.ER andabundantglycogenparticleisn cytoplasm.Fourteen(14)peroxisomesidentifieidnthiscell. Z 15715: Hepatocyte.Many dilatebdlebsofRER and abundantglycogenparticleisn cytoplasm.Mitochondriawithsome granuladrepositasnd dilatecdristaeN.ineteen(19) peroxisomesidentifieidnthiscell. Z15716: SinusoidwithliningendotheliacleH. Cellulafrixatioinspoor.Adjacent hepatocytecsontainmany mitochondriaR,ER and glycogen-likpearticlesD.ilatedblebs of RER arepresent.Multipleperoxisomesarepresentb,utnotcountedbecausewhole hepatocytesarenot inphotograph. Conclusions:Hepatocytesappeartocontainincreasednumbers ofperoxisomes,average 17/hepatocyt(e4cellscounted).Cellscontainabundantglycogen.Suboptimalcellular fixation. 418-014:PAGE F-23 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014:EM-99.46 Animal No.: 18952-2 Treatment Group: 1.6 mg/kg. high-dose Sex: non apiplicable Species/StraiCnr:l:CD(!)BRVAF/Plus(!D Rat Tissue: Lung Block No(s).: 99.46-18 Z15707 - Pboto/NegativeNo(s).: Z15711 SignificantLesions (check one): x Yes No InterpretingPathologist(signatureand date): Features: 15707: SinglePN2 liningalveolus.Cellcontainsmultipleriiitochondrairaound the nucleus,and alsomultipleclearvacuoles,which are mostly empty of contents.These were probably lainellabrodies which were extrudedintothe alveolarspace. These vacuoles are lined by a singlemembrane, whereas the mitochondriahave a double membrane. 15708: Collapsedalveolarsepta.A thinlayeredPN I coversthe surface.Beneath isan endothelialcellwith many pinocytoticvesicleso,ne a singlelamellarbody. Centrallyisan area of flattenedbasement membrane materialcontainingmany couagen fibrilsA. dark ceH ispresent intheupper right,which containsmany niitochondriaT.o the leftof thiscellare two lamellar bodies,although they appear tobe ina cytoplasmicextensionof an unidentifiedcell. Z15709: A nucleatedPN2 containingmultiplelamellarbodies(5)appearstobe overlainby a PN I liningthealveolus. Z 157 10: Alveolar space in top rightislinedby a thinPN I ceR. Adjacent to thisPN I and covered by it'scytoplasm isan extracellulaarreawithseveral(3)lamellarbodies. A capillaryis dorsalcontainingan erythrocyteand endothelialceU. Severalcellswithincollapsedseptaare not clearlyidentifiablaes to celltype. Z15711: A singledark PN2 issqueezed between two PNis. Itscytoplasm containsseveral empty vacuoles,which have apparentlyextrudedlamellarbodies. To the rightof thiscellare two largelamellarbodies withinsome lipid-likmeaterial.These bodiesmay be intheadjacent PN 1,macrophage, or may be extracellularA. smallcapillaryisinthelower rightmargin of the photograph. Conclusions: There appear to be increased numbers of type H pneumocytes (PN2s), and more lainellabrodies were seen in thissample. Otherwise,cellularmorphology appeared essentiallynormal. Fixationappeared satisfactory. 418-014:PAGE F-24 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-014:EM-99.46 Animal No.: 18956-1 TreatmentGroup: -1.6mg/kz.hieh-dose Sex: -- non ar)Rlicable Species/Stracirni:ocpebgvAF/pluse Rat Tissue: Liver BlockNo(s).: 99.46-13 Z15732 - Photo/NegativNeo(s).:Z15736 SignificaLnetsion(scheckone)X:- Yes No InterpretiPnagthologis(tsignaturaend date): Features:Z15732: Hepaticsinusoicdontaininngucleatederythroyctaend a couple matureerythrocytesS.ome lamellardebrisispresentinthesinusoidT.wo endotheliaclell nucleiarepresent.There areno completehepatocytesinthisphotograph. Z15733: Hepatocytecontainsabundantgranularparticlecsonsistenwtithglycogen,and numerous RER palisadesN.fitochondriaappeardenseand havesome granulardeposits anddilatedcristaeE.leven(11)peroxisomescounted.A bilecanaliculuisspresent between two hepatocytes. Z15734: Hepatocyte.SimilartoZ15733. Numerous dilatedblebsofRER arepresent. Twenty-four(24)peroxisomes. Z15735: Hepatocytewithsimilamrorphologyasdescribefdorabovecells.Sixteen(16) peroxisomescounted. Z15736: Hepatocyte.DilatedblebsofRER presenlNfitochondriaaredensewith granulardepositsand dilatecdristaeE.ighteen(18)peroxisomes. Conclusions:Increasednumbers ofperoxisomesperhepatocyte.17.25peroxisomes perhepatocyte(averageoffourcountedcells)S.uboptimalfixation. 418-014:PAGE F-25 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-014:EM-99.46 Animal No.: 18956-1 TreatmentGroup:-1.6mg&g. hilh-dose Sex:-non gpvlicable Species/StraiCnr:l:CMSBR VAF/Pluse Rat Tissue-:Lunp, BlockNo(s).: 99.46-14 Z15727 - Photo/NegativNeo(s).:Z15731 SignificanLtesions(checkone): x Yes No InterpretiPnagthologis(tsignaturaend date): Features:15727: Alveoluswithadjacenctollapsedsepta.The two bottompneumocytes arePN2 withlamellarbodies.Some ofthebodiesarepartialleympty,and inone cell appeartobe extrudingmateriaelxtracellulawriltyhotherlipidmaterial.At thetopisa largePN I.At theleftisa capillarwyitha nucleatederythrocyte. Z15728: Collapsedalveolasrepta.Severalcellscontainlamellabrodies.At leftone larnellabrody appearstobe withinthelumen ofa capillary. Z 15729: Somewhat T-shapedcapillarcyontaininagn electron-denseerythrocyteT.he endotheliaclellnucleusisventratlotheerythrocyteW.ithinthelumen ofthecapillary areseveralameuarbodies.At leftoffieldisthethincytoplasmiclininogfa PN I.The othercellsappeartobe sectiontshroughpneumocytesatvariousangles. Z 15730: Alveolarlumen and linincgells.Centralliysa smau PN2 containinag couple smalllamellabrodiesnexttoitsnucleus.The dark-stainicnygtoplasmtotherightofthis cellappearstobe an extensionfrom an alveolamracrophage.To theleftarea couple pneumocytes,and theycontaina couplelamellarbodies. Z15731:. Alveolarlumen and liningceus.TopleftceUisa PN2 withtwo lamellar bodies.A rightcentralp,ale-staininpgn,eumocyte alsocontainseveralsmalllarnellar bodies.The septaarecollapsedinthisarea. Conclusions: Increasednumber ofpneumocytescontaininlgamellarbodies.Some lameuar bodiesnotedin capillaries. 418-014:PAGE F-26 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.:-418-014,EM-99.46 Animal No.: 18973-1 TreatmentGroup:-1.6miz/kg.high-dose Sex:-non gpvlicable Species/StraiCnr:l:CDSBR VAF/Pluse Rat Tissue: Liver BlockNo(s).-:99.46-11 Z15742 - Photo/NegativNeo(s).:-1Z5747 SignificaLnetsions(checkone): x Yes No InterpretiPnagthologis(tsignaturaend date): Features:Z15742: Hepatocytecontainasbundantfinegranularmateriaclonsistenwtith glycogeninthecytoplasm.There arenumerous dilatebdlebsofRER on crosssection. Mitochondriaappeardense and have some granulardeposits.Seventeen(17) peroxisomescounted.Some lamellamraterianlotedextracellulaarnldy possiblywithin clearspacesin theleftportionof thehepatocyte. Z15743: Hepatocyte,overalslimilartoZ15742,althoughfixatioanppearsbetterN.ine (9)peroxisomescounted.Lamellarmateriaalttopappearstobe extracellular. Z15744: Hepatocyte.At leftisa hematopoieticcellinsinusoidandatupperright appearstobe a dilatebdilecanaliculus.Abundant areaofglycogenincytoplasm. Nineteen(19)peroxisomescounted. Z15745: Sinusoide,ndotheliaclelland incompleteportionsofa couplehepatocytes. Centralliysa clearcytoplasn-Lairceawithmany dilatebdlebsofRER. A few peroxisomesarepresent.No peroxisomecountmade. Z1546: Hepatocytecontainsabundantfinegranularmateriaclonsistenwtithglycogenin thecytoplasm.There arenumerous dilatebdlebsof RER on crosssection.Nfitochondria appeardense and have some granulardepositsand some dilatecdristaeE.leven (11) peroxisomescounted. Z15747: No photograph.Negativewas toodark.With fivephotographsalready evaluatedi,tisnotnecessarytoretakeortrytoprintthisimage. Conclusions: Appear tobe increasedperoxisomesperhepatocyte.Average 14/cel(lfour cellscounted.Suboptimalfixation. 418-014:PAGE F-27 TRANSNUSSION ELECTRON NUCROGRAPH INTERPRETATION StudyNo.: 418-014:EM-99.46 Animal No.: 18973-1 TreatmentGroup: 1.6mg/kizh,igh-dose Sex: non at)iplicable Species/StrainC:rl:CDOBR VAF/Plus(D Rat Tissue: Lung BlockNo(s).: 99.46-12 Z15737 - Photo/NegativeNo(s)Z.1:5741 SignificanLtesions(checkone): X Yes No InterpretiPnagthologis(tsignaturaend date): Features:Z15737: Collapsedalveolasrepta.Centralliysa PN2 withapproximately eightlamellarbodies,of which severalappeartobe releasintgheircontentsintothe extracellulsaprace.Ventrallayrealsosegmentsofa cell'csytoplasmthatappearsto containsome lamerarbody vacuoles.A couplePNIs arepresentadjacentothePN2, and a capillarwyitherythrocytaend endotheliaclellispresentinlowerleftcentralareaof photograph. Z15738: AlveoluswithlinincgellsT.he largenucleusbelongstoa PNl whose cytoplasmlinestheairspace.BeneaththisceU andjusttotherightareseverallamellar bodies,althoughit'sina subjacenctapillarTyo. themiddleleftisa capillarwyithan eyrthrocyttehatdoes containsome lamellardebrisinthelumen. Z15739: Alveolusand collapsesdepta.SimilartoZ15738,some lamellarbodiesappear tobe withinthelumen ofa capillaryP.NIs Linethealveolarspace. Z 15740: Largecapillarwyithseveralerythrocytetsotheright.Leftofthisisa cellular extensionp,robablya PN2, thatcontainsa distincltamellabrody. A thinlayerofPN I linesthealveolarspace. Z15741: AlveolarspacecontainsabundantlarnellamraterialS.everalPNls arepresent alongtheliningb,ut no PN2 arepresent.Lamellarbodiesarepresentinone capillary lumen and insome interstitsipaalcesintherightsideofthephotograph. Conclusions:Appear tobe increaseldamellarbodiesinthetissuecompared withcontrol sample,althoughmany were incapillarlyuminaand some inalveolarand interstitial spaces. 418-014:PAGE F-28 M. TransmissionElectronNficrographs 418-014:PAGE F-29 NOTE: The originagllossyfigurpehotographs(FiguresI-12)were deliveretdo Dr.Marvin T.Case withthedraftpathologyreporton March 19,1999. 418-014:PAGE F-3C) AncillaryPathology Rcport Primedica Argus Study No. 418-014 FigureLegends Figure1.Liver.HepatocytsehowingnormalcytoplasmiocrganefleWsh.itespotswithin rriitochondrrieapresenstome dilatatiofncristaeS.ome peroxisomesareidentifiedT.o therightisan endotheliaclelland erythrocyteosfadjacenstinusoid.Animal No. 189041,0 mg/kg/day(control)1,1,20OXmagnification. Figure2. Liver.Hepatocyte.Dilatatioonfnuclearenvelopeandroughendoplasmic reticulum(blebbingi)ndicatessuboptimalfixationM.tochondriaaredenseand show some granulardepositsand dilatedcristaeS.ome representatipveroxisomesarelabeled. Hematopoieticcellsarepresentinsinusoidatupperrightofphotograph.Animal No. 18912-1,0 mg/kg/day(control)I,1,20OXmagnification. Fizure3. Liver.CellulamrorphologyissimilatroFigure2. Some peroxisomesare labeleda,nd appearmore numerous thanincontrols.A bilecanaliculuisspresentin upper rightofphotographand hematopoieticcellsarepresentinsinusoidatupperleft. Animal No. 18952-2,1.6mg/kg/day(high-dose)I,1,20OXmagnification. Figure4. Liver.Dilatatioonfroughendoplasmicreticulumisevident.Some peroxisomes,increasedinnumber,arelabeled.Animal No. 18956-1,1.6mg/kg/day (high-dose)1,1,20OXmagnification. Figure5. Liver.Hepatocytedemonstratinsgomewhat bettefrixatiownithmore even dispersioonforgancllesC.learspaceattopofcellw,hich containsome membranous debrisa,ppearstobe extracellulianrtheperisinusoidsaplace.Animal No. 18973-1,1.6 mg/kg/day(high-dose)I,1,20OXmagnification. Figure6. Lung. Type IIpneumocyteispresentalongthealveolawrall,adjacentotypeI pneumocyte. Lamellarbodieswithinthecytoplasmidentiftyhiscellsasa typeIl pneumocytc. Animal No. 18904-1,0 mg/kg/day(control1)5,,680Xmagnification. Figure7. Lung. Threecuboidalpneumocyteslinethisairwayspace.A flattenetdypeI pneumocyte isevidentalongtheventraplortionoftheairwayspace.Two smalllamellar bodiesinone cuboidalcellsuggestthisisa type11pneumocyte which has secretedmost of its lainellar bodies and isdestinedtobecome a typeIpneumocyte. Animal No. 189042,0 mg/kg/day(control)1,1,20OXmagnification. Figure8. Lung. A more dcnsely-stainitnygpe]Ipneumocyte,containintgwo lamellar bodies,appearssqueezedbetweentwo typeI pneumocytes.Some lamellarmaterialis alsopresentwithinthecapillarlyumen oftheseptalwallbothabove and totherightof thetypeIlpneumocyte. AnimalNo. 18915-1,0 mg/kg/day(control1)1,,20OX magnification. 418-014:PAGE F-31 AncillaryPathology Report Primedica Argus Study No. 4 18-014 Figure9. Lung. Dense-stainincgellisa type11pneumocytepositionebdetweentwo type Ipneumocytes.Four distincltamellarbodiesarepresentinthecytoplasmof thetypen pneumocyte,althoughthelameflarmaterialispartiallwyashed out inthesevacuoles.To therightofthiscellaretwo darklameflar-rybpoediessurroundedby some lipid.This appearstobe withina cellina collapsedareaofsepta,and may be a macrophage. Animal No. 18952-2,1.6mg/kg/day(high-dose)1,1,20OXmagnification. Figure10.Lung. Collapsedareaofseptashowingtwo typeIIpneumocytescontaining lamellabrodies.A typeI pneumocyteistotheleftofthesecells.Capillarieasnd adjacent alveolacrellsarepresentdorsallaynd totherightofthephotograph.Animal No. 189521,1.6mg/kg/day(high-dose)1,1,20OXmagnification. Figure11.Lung. Centralliysa cellcontaininsgeveralamellar-liskteructurewshich appearstobe a type11pneumocyte.Glycogen-likmeateriaalppearstofilmluch ofthe cytoplasm.Adjacentceus totherightcontaincentrioleasnd represenctiliated bronchiolacrells.Cuboidalepithelicaelllsaretotheleftand ventratlothetypeH pneumocyte. Animal No. 18952-1,1.6mg/kg/day(high-dose)1,1,20OXmagnification. Figure12.Lung. Alveolarspacecontainsabundantlamellar-limkaeterialw,hich is consistenwtithsurfactanstecretedfrom typeH pneumocytes.A typeIpneumocyte lines thespaceon theleftand rightT.wo cuboidalcellcsontaininglycogen-likgeranular materialappeartobe pneumocytes,probablytypeIIswhich havereleasedtheirlamellar bodies.Animal No. 18973-1,1.6mg/kg/day(high-dose)1,1,20OXmagnification. 418-014:PAGE F-32 AncillaryPathology Report Primedica Argus Study No. 418-014 IndexofLabelsforUltrastructuSrtarluctures Abbreviation Structure A Alveolarspace BC Bilecanaliculus BE Bronchiolaerpithelium CL Capillarlyumen Cap Capillary CN Centrioles CP Cuboidalpneumocyte D Desmosome DD Dense deposits DC Dilatedcristae DN Dilatednuclearenvelope DR DilatedRER EC Endotheliaclell ER Erythrocyte G Glycogen HC Hematopoieticells L Lipiddroplet LB Lamellarbody m NEtochondria N Nucleus p Peroxisome PN1 Type Ipneumocyte PN2 Type IIpneumocyte PM Plasma membrane RER Rough endoplasrrdcreticulum 418-014:PAGE F-33 -P '.DC EiC' 418-014:PAGE F-34 ell HC 'DD D .1 lp 418-014:PAGE F-1-35 G DR @N -fez.-. D14 DR 418-014:PAGE F-36 sk RER, AP ,L. ib 67 lot- IL r lit VP.- 418-014:PAGE F-37 'E IV lot Is*. L 14 CL 39 418-014:PAGE F-38 4r Ir A Nl . ir- 418-014:PAGE F-39 !Pic SUL Jr. A dr PNI '41 -A 'k 418-014:PAGE F-40 L 63 418-014:PAGE F-41 'LLNd op -lk 418-014:PAGE F-42 FrA ap PNi I'@ f4,h ' IL let CA 4 18-014:PAGE F-43 .0w CN CN tit.. Yr ntW.. JA@ ziv@ Lo fv 4:@4* Ob 418-014:PAGE F-45 IV. QualityAssuranceStatement AL-ffff-ii 418-014:PAGE F-46 Pathology Associates international A Company of Science ApplicationsIrltemationaClorporation "=6"0,0"0-@wdq@o QUALITY ASSURANCE STATEMENT Thiselectronmicroscopystudyhas been inspectedand auditedby theQuality Assurance Unitas requiredby theGood LaboratoryPracticesRegulationspromulgated by theU.S.Food and Drug AdministratioPnA.I has a functioninagnd responsive QualityAssurance Unitwhichreportsdirectltyomanagement. The followinigsa record ofinspections/audiatnsd theiresultinrgeports: Date of lnspect*on Phase Inspected Date FindonasRel2orted toManagement and StudyDirector 2/18,19/99 3/16/99 3/17/99 4/5/99 Processingand Embedding Data Audit DraftReportAudit FinalReportAudit 2119/99 3/17/99 3/17/99 4/5/99 n deward, B.S.,RQAP-GLP alitAyssurance Specialist Z,<Z:r U-afe Sponsor: 3M CorporateToxicology TestArticleP:FOS StudyNumber: 418-014,PAIStudyEM-99.46 4915D ProspectusDriveo Durham,NorthCarolina277139 (919)544-5257 (919)544-3218FAX STATEMENT APPENDIX G OF THE STUDY DIRECTOR iriPMMEDIC.,k *18-014P:AGE G-I Argus Resear-chLaboratories.Inc. 905 Sheehy Drive.BuildingA Horsham, PA 19044 Telephone:(215)443-8710 Telefax:(215)443-8587 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS SPONSOR'S STUDY NUMBER: T-6295.13 STATEMENT OF THE STUDY DIRECTOR Thisfinalreportaccuratelyreflecttshe raw data obtainedduringthe performance ofthe study. No deviationsfrom the U.S. Food and Drug Administratio(nFDA) Good LaboratoryPracticeRegulations;FinalRulea ,the Japanese Ministryof Healthand Welfare(MHW) Good LaboratoryPracticeStandard forSafety Studieson Drugs' and the European Economic Community (EEC) Council decisionon 28 July 1989 on the acceptance by the European Economic Community of an OECD decisionlrecommendationon compliance withprinciples ofgood laboratorypracticeocccurredthataffectedthe qualityor integritoyfthe study. 4.tE - ---------------------k@4nt;!@nd-G@.. Y@orrk, .,DABT AssociateDirectorof Research and Study Director 99 Date ---------------------- a. U.S. Food and Drug Administration.Good LaboratoryPractice Regulations;FinalRule. 21 CFR Part58. b. Japanese Ministryof Health and Welfare (1988). Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. C. European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decisionlrecommendation on compliance with principles of good laboratory practice. Official Journal of the European Communities: Legislation3.2(No.L 315; 28 October):1-17. APPENDIX H QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT iriPMMEDIC,-A, 418-014:PAGE H-1 Argus Researrh Laboratories,Inc. 905 Sheehy Drive.BuildingA Horsham. PA 19044 Telephone: (215)443-87 10 Telefax:(215)443-8587 QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT Study Director:Raymond G. York,Ph.D.,DABT ExecutiveDirectorof Research: MildredS. ChristianP,h.D.,Fellow,ATS Protocol418-014: Oral(Gavage) Cross-FostedngStudy ofPFOS inRats Sponsors Study Number: T-6295.13 The draftprotocolforthisstudywas auditedforadherence to U.S. Food and Drug Administratio(nFDA) Good LaboratoryPracticeRegulations,Japanese Ministryof Healthand Welfare (MHW); Good LaboratoryPracticeStandard for SafetyStudieson Drugs, and European Economic Community (1989)council decisionon 28 July1989 on the acceptance by the European Economic Community ofan OECD decision/recommendationon compliance withprinciples of good laboratorypracticeon 19 OCT 98. Criticaplhases ofthisstudywere inspectedninetimes;study information and raw data were auditedtwice(see tables1 and 2 fordates and phases/data). The draftfinalreportand the raw data forthisstudy were compared and auditedforaccuracy,foradherence to protocolrequirements,and foradherence to U.S. Food and Drug Administratio(nFDA) Good LaboratoryPractice Regulations,Japanese Ministryof Healthand Welfare(MHW); Good Laboratory PracticeStandard forSafetyStudieson Drugs, and European Economic Community (1989)councildecisionon 28 July1989 on theacceptance by the European Economic Community ofan OECD decision/recommendationon compliance withprincipleosfgood laboratorypracticebetween 21 MAY 99 and 10 JUN 99, and forrevisionsrequestedby the Sponsor on 20 JUL 99, 22 JUL 99, and forfinalizatioonn 23 JUL 99. 418-014:PAGE H-2 Thisstudy was conducted accordingto U.S. Food and Drug Administratio(nFDA) Good LaboratoryPracticeRegulations,Japanese Ministry ofHealthand Welfare (MHW); Good LaboratoryPracticeStandard forSafety Studieson Drugs,and European Economic Community (1989)councildecision on 28 July1989 on the acceptance by the European Economic Community ofan OECD decision/recommendationon compliance withpdnciplesofgood laboratorpyractice. 16, Nancy 0ng 1 -wski QualityAssurance Manager Date Heather L.RaVuttino,M.S. Date QualityAssurance Supervisor and PrincipaAluditor TABLE 1 CRITICAL PHASES INSPECTED 418-014:PAGE H-3 Test ArticlAedministration-Gavaae Date of inspection:03 NOV 98 Date resultsreportedto the Study Directorand Management: 06 NOV 98 Test ArticlePrei)aration Date ofinspection:03 NOV 98 Date resultsreportedtothe Study Directorand Management: 06 NOV 98 Cohabitation Date of inspection:17 DEC 98 Date resultsreportedtothe Study Directorand Management: 18 DEC 98 NaturalDelivery/LiftOebrservations Date of inspection:06 JAN 99 Date resultsreportedtothe Study Directorand Management: 19 JAN 99 Blood Collectio-n Dams and Pur)s Dates of inspection:28 JAN 99, 28 JAN 99 Dates resultsreportedto the Study Directorand Management: 02 FEB 99,02 FEB 99 Fecal and Urine Collection 418-014:PAGE H-4 Date of inspection:28 JAN 99 Date resultsreportedtothe Study Directorand Management: 02 FEB 99 Scheduled Sacrific-eDams and Pur)s Dates of inspection:28 JAN 99,28 JAN 99 Dates resultsreportedtothe Study Directorand Management: 02 FEB 99,02 FEB 99 418-014:PAGE H-5 TABLE 2 RAW DATA AUDIT(S) The followingstudy informationand raw data were audited from 26 FEB 99 to 10 MAR 99: Animal receiptr,andomization,physicalexaminationand In-lifteransactionrecord. Feed consumption. Cohabitation. Naturaldeliveryobservations. Lifteorbservations. Pup body weights and status. Table of random units. Milkcollectiodnata and packing lists. Blood and livecrollectiodnata and packing lists. Urine and fecalcollectiodnata and packing lists. Necropsy. Tissue packing lists. Male breeder colonyrecords. General comments. Study maintenance records. Temperature and relativheumidityreports. Feed, water and bedding analyses. Editrequests. Dosage volumes. Data review pages. acclimation. The resultsofthisauditwere reportedtothe Study Directorand Management on 12 MAR 99. The followingstudy informationand raw data were audited from 26 FEB 99 to 10 MAR 99: Vehiclereceiptp,reparationand use. Test articlreeceipt,preparationand use. Test articlpeacking lists. The resultsofthisauditwere reportedtothe Study Directorand Management on 12 MAR 99. PathologyAssociateslntemational A Company ofScienceApplicatioInnsternatioCnoarlporation An Emplovee-Ovoed CCmoan@ PATHOLOGY REPORT (ANCILLARY STUDY) ELECTRON MICROSCOPIC EVALUATION OF LIVER AND LUNG AND LIGHT MICROSCOPIC EVALUATION OF LIVER IN CRL:CDOBR VAF/PLUS@ RATS) ORAL (GAVAGE) PRARMACOKINETIC STUDY OF PFOS IN RATS CLIENT STUDY NUMBER 418-013 PAI STUDY NUMBER 99-11(EM 99.39) Prepared by: James B.Nold,D.V.M.,Ph.D.,Diplomate,A.C.V.P. PathologyAssociatesInternational 4915-D ProspectusDrive Durham, NC 27713 April5, 1999 PREPARED FOR 3M CORPORATE TOXICOLOGY, 3M CENTER, ST. PAUL, MN 55144-1000 4915 D ProspectusDriveo Durham, NorthCarolina27713 o (919)544-5257o (919)544-3218FAX PathologyNarrative LightMicroscopy and TransmissionElectron Micrograph InterpretatiFoonrms TransmissionElectronMicrographs QualityAssurance Statement Section I H III IV I. PathologyNarrative PATHOLOGY (ANCILLARY REPORT STUDY) AnciiiaT-NP@athology Report PrimedicaArgus No. 418-013 ELECTRON MICROSCOPIC EVALUATION OF LIVER AND LUNG AND LIGHT MICROSCOPIC EVALUATION OF LIVER IN CRL:CD@BR VAF/PLUS@ RATS) ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS CLIENT STUDY NUMEBER 418-013 PAI STUDY NUMEBER 99-11 (EM 99.39) SUND4MARY Ultrastructureavlaluationof liverand lungsofratpups from dams treatedwith 0 (control)0,.4,1.6,or 3.2mg/kg/day (high-dose)PFOS identifieidncreasednumbers of peroxisomeswithinhepatocytesof high-doseratpup livers.Quantitationof peroxisomes indicatedatleasttwice thenumber ofperoxisomes inanimalstreatedwith 0.4 mg/kg/day and tripltehe number inthosegiven 1.6or 3.2 mg/kg/day. Differencesincellular membranes or n-@tochondriawere notdiscerniblebetween controland treatedanimals in the samples examined; however, suboptimalfixationprecludeda detailedevaluationof theseorganelles.Inlung samples,thepresenceof typeIIpneumocytes and lamellar bodieswas evaluated.No significanutltrastructurdailfferencewsere identifiebdetween lung from controland treatedrats.Random type11pneumocytes containinglamellar bodieswere identifieidnalllungsexamined ultrastructurally. PROCEDURES The objectiveof thisstudywas toexamine by electronmicroscopy theliversand lungs from selectedday-21 gestationratfetusesinan oral(gavage)pharmacokineticstudyof PFOS. In additiontoa generalqualitativeevaluationfordifferencesbetween controland treatedanimals,theultrastructureanldpointsincludedquantitatioonf peroxisomes within hepatocytes,membrane and mitochondrialchanges inhepatocytes,and thequantitatioonf surfactanltamellarbodiesinthe lung.Lightmicroscopyof paraffin-embeddedliver sectionswas alsoperformed. For the pharmacokineticstudy,90 female Crl:CD@BR VAF/PLUS@ ratswere given the testarticloer thevehicleonce dailybeginning42 days priortocohabitationwith a male through day 20 postpartumaccordingtoText Table 1. Dosage Group IV v AncillaryPathologyReport PrimedicaArgus No. 418-013 Text Table 1. Dosage Levels,Concentrationsand Volumes Number of Mated Female Rats 16 16 1 16 16 PFOS Dosage (mg/kg/day) 0 (Vehicle) 0.1 0.4 1.6 3.2 PFOS Concentration (MGJML) 0 0.02 0.08 0.32 0.64 Dosage Volume (mIJkg) 5 5 5 5 5 At Caesarean-sectioning on day 21 of presumed gestation, lung and liver samples were collectedfrom three fetusesper littefrrom five litterpser dosage group. Samples of lung and liverfrom selectedfetusesforGroups I,IH, FV, and V were thin-slicedand placed in McDowell-Trump fixative(McDowell EM, 1976) and submitted to thislaboratoryfor electronmicroscopy processing and evaluation. Sections of liverfrom these same animals were irnmersion-fixedin neutralbuffered formalin and submitted along with electronmicroscopy samples. Per sponsor request, tissuesamples from selectedpups were processed and evaluated (Text Table 2). The fivedigitnumber isthe dam number; the suffixes1-4 denotes the pup number from thatdam's litter. Text Table 2. Animals SelectedforElectronMicroscopy Group I Control(0mg/kg/day) 19504-3 19505-1 19506-4 19509-1 19516-2 1 Group III 0.4 mg/kg/day 19534-1 19536-3 19543-4 19546-1 19546-2 Group IV 1.6mg/kg/day 19560-1 19560-2 19560-3 19562-2 19562-3 Group V 3.2 mg/kg/day 19568-2 19569-1 19572-1 19577-4 19579-3 The lungs and liverswere then processed intoSpurr's resinforultrastructural examination. Thick (1g) sectionswere cut,stainedwith toluidineblue,and examined to selectrepresentativeareas forfurtherelectronmicroscopy processing. Thin sections (approximately 90 nm) were cut,mounted on 200-mesh copper grids,stainedwith 5% methanolic uranyl acetateand Reynold's lead citrate,and examined on a Zeiss 900 transmission electron microscope. Centrilobularhepatocytes, where clearlyidentifiable in liversections,were preferentiallyexamined. Representative electron photomicrographs of liverand lung were taken and significantultrastructurafleatures were summarized for each photograph and animal on a designated transmission electron micrograph interpretationform. The number of peroxisomes in hepatocytes was manually counted for each centerphotographed hepatocyte and recorded. For lightmicroscopy, formalin-fixedliversamples were processed and embedded in paraffin,cut at approximately fivemicrons, stainedwith hematoxylin and eosin,and evaluated. 1-2 AncillaryPathology Report PrimedicaArgus No. 418-013 RESULTS AND DISCUSSION Individualinterpretatioonfslightmicroscopy slidesand electronmicrographsforeach animal are shown in SectionII.Selectedrepresentativeelectronphotomicrographswere labeledand are shown in Section1111. Lii!htMicroscopy A summary of the incidenceand severityof lightmicroscopic findingsin the liveris shown inText Table 3. There was an increasedincidenceand severityof eosinophilic granularitoyf thecytoplasm of hepatocytesintreatedratscompared with controlrat livers.The severityof thisfindingincreasedina dose-relatefdashion.Hepatocellular eosinophiligcranularitwyas diffuselydistributeidnmost animalsgiven 1.6or 3.2 mg/kg/day of PFOS, but was predominantlyperiportailn I animalgiven 1.6and in3 animals given 0.4 mg/kg/day. Cytoplasmicclearingofhepatocyteswas prominent in controllivers.This morphologicaldescriptioinsusuallyassociatedwith the normal accumulationof glycogen inhepatocytes.In thisstudy,theincidenceand severityof cytoplasmicclearingwas almostinverselyproportionaltotheoccurrenceof eosinophilic gDranularity. The liverisa hematopoieticorgan inratfetusesand neonates.Thus, largenumbers of hematopoieticcellsarepresentwithinthesinusoidsthroughouttheliver.Extramedullary hematopoiesis(EMH) occurredinallanimals,includingcontrolsa,nd was uniformly graded as moderate tomoderately-severe.The severityof EMH was increasedinthe treatedgroups. Random mitosesof hepatocytesarenormal inratliverse,speciallyin fetusesand neonates.There appeared tobe a slightincreaseinhepatocellulamritotic figuresinthe animalsgiven 3.2mg/kg/day PFOS. Text Table 3. Incidenceand Severityof LightMicroscopicLiverFindin-s Microscopic Finding (numberexamined) Eosinophiglriacnularciyttyo,plasmic, increased minimal slight moderate Cytoplasmic clearing minimal slight moderate Extramedullaryhematopoiesis moderate moderateIy-severe Mitotic fi-ures,increased minimal slicyht PFOS Dose Levels(mg/kg/day) 0 0.4 1.6 3.2 (5) (5) (5) (5) 2 2 1 - - 3 3 1 1 4 1 2 - - 4 1 5 - I 5 4 1 - - 1 4 5 2 1 2 2 - - 2 Electron Microscog)v AncillarPyathologRyepon PrimedicAargusNo. 418-013 Fixatiofnthetissuweass lessthan optimal forelectronmicroscopy. This was especially evident in the livercompared with the lung. Lung isinherentlylessdense than liverand allows betterinfiltratioonf the fixativeinto the tissue.Suboptimal fixationwas characterizedby clumping of largeorganelles,discontinuous plasma membranes, loss of detailof cytoplasmic organelles,dilatationof endoplasmic reticulum and the nuclear envelope, granular deposits in the mitochondrial matrix, and dilatationof mitochondrial cristae.The poor fixationof membranes made ultrastructuraalssessment of these structuresimpossible. Fixation,however, was stilclonsidered adequate to assess forthe presence of peroxisomes in the hepatocytes and lamellar bodies in the lung and to make general comparisons between control and treatedsamples. LIVER Control (0 mg/kg/day). Subcellularfeaturesof controlhepatocytesare demonstrated in Figures I and 2. Hepatocytes characteristicallcyontain numerous mitochondria and prominent endoplasmic reticulum. Clumping of the large organelles(n-titochondria, rough endoplasmic reticulum and peroxisomes) isindicativeof poor fixation.In these cellsrough endoplasmic reticulum was evident,but smooth endoplasmic reticulum was not because of suboptimal fixation.Mitochondria generally appeared dark and contained scattereddense deposits and some dilatedcristae,both artifactsdue to suboptimal fixation.Dilatationof endoplasmic reticulum and the nuclear envelope were additional artifactosf poor fixation.The fine granular materialin the cytoplasm isconsistentwith glycogen particles.Peroxisomes were identifiedand quantitatedin from representative hepatocytes. For the control livers,the average number of peroxisomes counted per hepatocyte photographed was 4.2 peroxisomes per cell(Text Table 4). Fetal and neonatal ratliverisa hematopoietic tissue,and numerous hematopoietic cellswere evident within the liversinusoids. Text Table 4. Quantitationof HepatocellularPeroxisomes 0 Animal Number 19504-3 19505-1 19506-4 19509-1 19516-2 Mean = Average g/kg Number' of Peroxisomes 3.2 3.0 4.4 5.2 5.0 4.2 0.4mg/kg Animal Number 19534-1 19536-3 19543-4 19546-1 19546-2 Number of Peroxisomes 1-2.0 12.2 8.0 13.0 5.4 Mean = 10.1 I.. mg/kg Animal Number Number of Peroxisomes 19560-1 11.6 19560-2 12.8 19560-3 10.2 19562-2 10.6 19562-3 20.0 Mean = 13.0 3.2 mg/kg Animal Number of Number Peroxisomes 19568-2 6.0 19569-1 10.6 19572-1 13.2 19577-4 11.2 19579-3 21.8 1 Mean 12.6--1 number of peroxisomes per hepatocyteforfivehepatocytescounted per animal. 0.4 mg/kg/day. The average number of peroxisomes per hepatocyte (10.1) was higher in the animals given 0.4 mg/kg/day PFOS compared with controls(Text Table 4). A representativehepatocyte isshown in Figure 3. Qualitatively,therewere no identifiable 1-4 AncillaryPathology Report PrimedicaArgus No. 418-013 ultrastructurdailfferencebsetween the hepatocytestreatedwith0 (control)and 0.4 mg/kg/day. 1.6mg/kg/day. For animalsgiven 1.6mg/kg/day,theaveragenumber of peroxisomes per hepatocytewas 13.0.Figure4 illustrataesrepresentativheepatocytefrom this treatmentgroup. Otherwise,generalultrastructurmaolrphology was similarinhigh-dose liverscompared with controllivers. High-Dose(3.2mgtkg/day). The averagenumber ofperoxisomes per hepatocyte(12.6) was higherinthehigh-doseanimalscompared withcontrols(Text Table4). Figures5 and 6 illustrartepresentativheepatocytesfrom high-doseanimals.Differencesin mitochondriaand cellulamrembranes were notdiscerniblbeetween treatedand control livers.Suboptimal fixationh,owever, ofthe tissuesprecludeddetailedevaluationof these organelles. LLTNG 0 mgtkg/day (Control). Lung samples were atelectatiacnd not inflatedwith fixative. Therefore,most alveoliand alveolarseptawere collapsedwith poor delineatioonf alveolarand airway spaces.Figures7-9 illustrastoeme normal cellularitaynd ultrastructurpaulmonary anatomy. Type 11pneumocytes (PN2) were randomly identified liningalveolit,erminalbronchioleso,r withincollapsedsepta.Type IIpneumocytes were identifiebdy thepresenceoflamellarbodies,which arecytoplasmicvacuolescontaining electron-densleaniinatemdembrane-like profiles.Lamellarbodiesrepresentthe material secretedintothealveolarspaces as surfactantO.ccasionallysome lamellarbody vacuoles were empty, as ifthe lamellarmaterialhad washed out or been secreted.Surfactantor lamellarmaterialwithinairway spaceswas only occasionallyidentifieidncontrollungs or treatedlungs.The typeI pneumocyte (PN I)ischaracteristicaflllayttenedand linesthe alveolarspace as a thinmargin of cytoplasm. Type I pneumocytes differentiatferom type Ilpneumocytes. Many pneumocytes appearedtobe cuboidalwithoutdistinctivleamellar bodies,and may representa transitiosntagebetween PN2s and PN Is,especiallysince theselungs were fetaland had not been expanded withair.Additionallys,ome cuboidal epitheliaclellsrepresentedterminalbronchiolarepithelium. Type IIpneumocytes and lamellarbodieswere randomly identifieidnthesecontrollungs. Some lamellar-likbeodieswere alsopresentwithincapillarlyumina,althoughinthe controlsectionsexamined no lamellarbodieswere clearlyidentifieidnthealveolar lumina. 0.4 mg/kg/day. Qualitativelyt,helung samples from animalsgiven 0.4 mg/kg/day appeared essentialltyhe same ascontrolsamples. A representativseample isshown in Figure 10. Random typeIIpneumocytes containinglamellarbodieswere identifieda,nd only occasionallysome lamellarmaterialwas seen withinalveolarlumina. 1-5 Anciiiar,P.athologyReport PrimedicaArgus No. 418-013 1.6mgtkg/day. FiguresII and 12 illustrastoeme representativueltrastructurfaelatures from ratfetalung samples given 1.6mg/kg/day. These were consideredessentially similartothecontroland othertreatedsamples. Lamellar bodieswere identifiewdithin typeH pneumocytes and,occasionallyw,ithinalveoli. 3.2 mg/kg/day (High-Dose). Ultrastructurpaalthologyof thehigh-doselungs was essentiallsyimilartocontrol,low- and mid-dose samples. The collapsednatureof most of thealveoliand septamade definitivqeuantitatioinmpracticalS.ome lamellarbodies were identijfiweidthincapillarlyumina and only a smallamount was seen withinalveoli. Figures 13 and 14 illustrastoeme representativleungfieldsfrom the high-doseanimals. CONCLUSION Ultrastructureavlaluationof liverand lungs ofratpups from dams treatedwith 0 (control)0,.4,1.6,or 3.2mg/kg/day (high-dose)PFOS identifieidncreasednumbers of peroxisomes withinhepatocytesof ratpup liversfrom dams given 1.6or 3,2 mg/kg/day. Quantitationof peroxisomesindicatedatleasttwice thenumber ofperoxisomes in animals treatedwith 0.4 mg/kg/day and triplethenumber inthosegiven 1.6or 3.2 mg/kg/day. Differencesincellulamrembranes or mitochondriawere not discernible between controland treatedanimalsin thesamples examined; however, suboptimal fixationprecludeda detailedevaluationoftheseorganelles. In lung samples,thepresenceof typeIlpneumocytes and lamellarbodies was evaluated. No significanutltrastructurdailfferencewsere identifiebdetween lung from controland treatedrats.Random typeIIpneumocytes containinglamellarbodieswere identifieidn alllungsexamined ultrastructuralAllyt.hough some lamellarmaterial(surfactantw)as morphologicallyidentifiablweithinalveolarlumina,no significandtifferencewsere noted between controland treatedsamples. SIGNATURE OF AUTHOR @ ,v-@eQ-@ @-@amesB. Nold, D.V.M., Ph.D., Diplomate,A.C.V.P. Date' REFERENCES McDowell, EM and Trump, BF: Histologicfixativfeorroutinediagnosticlightand electronmicroscopy.Arch. Pathol.Lab. Med., 100:405-414,1976. 1-6 11.Light Microscopy and TransmissionElectron Micrograph InterpretatioFnorms 1 I Forms aretypographicallcyorrectedversionsof signe(i/daterdaw datasheetsmaintained in archivedstudy file. LIGHT NHCROSCOPIC EVALUATION Study No.: 418-013 (PAI 99-111 Species/StrainC:rl:CD@BR VAF/Plus@ Rats,Term Fetuses Animal No: seebelow Sex: -n/a InterpretinPgathologis(tsignaturaend date): Tissue: Liver Animal Dose Number MicroscopicFindingsand Comments 0 mg/kg 19504-3 1. Extramedullarhyematopoiesi(sEMH), diffusem,oderate 2. Cytoplasmiclearingd,iffusem,oderate .............................i....@.:..n.@.g.t...i..c...f...i..g..u...r.e.s.m.,i.n.i..m.a.l.,.m.u.l.t..i.f.o.c.a..l........................................................... 19505-1 1. EMH, diffusem,oderate ...........................................q.y...@g.pl.a.s.m.i.c..cl.e.a.r.i.n..g..,..d..i..f..f..u..s..e.,...m..o..d..e..ra.t.e........................................................... 19506-4 1. EMH, diffusem,oderate 2. Cytoplasmicclearingd,iffusem,oderate ...............................3....E..oW.s..i.@n.op.h.i..l.i.g.c.r.a.n..u.laritcyy,.!pIpMTi.c.,......i.n.rce..a.s.e.d..,p.e.r..i.p.o.r..t.a.ml.i,.n.i..m.a.l............................. 19509-1 E iffusme;,@@ 2. Cytoplasmicclearingd,iffusem,oderate ...............................3....E..o.sino.p.h.i..l..ig.c.r.a.n.u.l..a.r...i..tc...y..,!..g..p..1.@@!.@.!..i..c..,.r.e.ia.ns.c.e.d.,..p..e..r..i..p.o.r.t.a.l..m.,.i..n...i..m..a..l..................... 19516-2 1. EMH, diffusem,oderate 2. Cytoplasmicclearingd,iffusem,oderate 3. Mitotifciguresm,inimal,multifocal 0.4mg/kg 19534-1 1.EMH, diffusem,oderate 2. Cytoplasmicclearingd,iffusem,inimal ...................................@..o.s...i..!..o!..p.h..i.l.i.cg..r..a..n..u..l...a..r.i.t.y,.@@.y.@.p.p.l.a..si.mn.ic.cr.,e..a.s.e.d.d,.i.f.f.u.s..e.m,.i.n.i.m.a.l................................... 19536-3 1.EMH, diffusem,oderate 2. Cytoplasmicclearingd,iffuses,light .........................................3...E..o.s.i.n.o..p...h..i...l..i..gc..r...a..n...u.l.a.r.i..!.y....,...c.y..@..p..p...l..a.i.sn.m.ci.rc.e,.a.s.e.d.p,.e.r...i...p...o.r.t..a..l.s..,.l..i..e..-...@..!................. 19543-4 1.EMH, diffusem,oderate 2. Cytoplasmiclearingd,iffuses,light .............................................................. ...y....c.y.@gp l..a.s.m.i.c..,i.n.c.r..e.a.s.e.d.p.,@..T.i.P.o..r..t..a..,......s...l...g.......@..@...................... 19546-1 1. EMH, diffusem,oderate 2. Cytoplasmicclearingd,iffuses,light ..................i..3....E.o.s.i.n..o..p...h..i..l...i.g..c.r...a..n...u..l..a..r.y.i....t...@..@....y9...1tp...l..a.s..m.i.c.,i.n..c.r.e.a.s..e.dd.,i..f.f.u.s.e.m.,i.n..i.m.a.l.................................. 19546-2 1. EMH, diffusem,oderatey]-severe 2. Cytoplasmicclearingd,iffuses,light 3. Eosinophiligcranularitcyy,toplasmici,ncreasedp,eriportasll,ight 4. Mitoticfiguresm,inimal,multifocal Page Iof 3 Animal Dose Number MicroscopicFindingsand Comments 1.6mg/kg 19560-1 1. EosinophiIicgranularityc,ytoplasmic,increased,moderate, diffuse .................................2....E..M.H.,...d..i.f.f.u.s..e.m,.o.d..e.r.a.t..el.y...s..e...v...e..r..e......................................................................................... 19560-2 1. EMH, diffusem,oderately-severe 2. Cytoplasmic clearingd,iffusem,inimal 3. Eosinophilicgranularityc,ytoplasmic,increasedd,iffuse,slight ..................................4.........M...i..t..o...t...i..c.f..i.gY..r..e...s..,.m...i..n...i..m...a..l..,.m..u.l.t.i..f.o.c.a.l................................................................. 19560-3 1. EMH, diffusem,oderate 2. Cytoplasmic clearingd,iffusem,oderate 19562-2 3. Eosinophilicgranularityc,ytoplasmic,increasedp,eriportal,minimal n @gtic f.i.g.u.r..e.s.m,.i.n..i.m.a.l.,..m.u.l.t.i..f.o.c.a..l................................................................ 1 . EMH, diffusem,oderately-severe 2. Cytoplasmic clearingd,iffuses,light ..l.@@@inophilicgranulari!._y@,ylp.p1@@Tic increased,diffuse,slight .....................4...............................I......... .................................. ............................................................................... 19562-3 1. EMH, diffusem,oderately-scvere 2. Cytoplasmic clearingd,iffusem,inimal 3. Eosinophilicgranularityc,ytoplasmic,increasedd,iffuse,slight 3.2 mg/kg 19568-2 1 . Eosinophilicgranularityc,ytoplasmic,increased,moderate, diffuse 2. EMH, diffusem,oderately-severe .....................................M.@.g.t.i...c.....f.i.g.u..r.e.s.s,.l..i.g..h...t..m,..u..l..t.i.f..o.c.a.l................................................................... 19569-1 1. Eosinophilicgranularityc,ytoplasmic,increased,slight diffuse 2. EMH, diffuse,moderately-severe ......................-.-.-.-.-.-.--. 19572-1 ..3....M..i.t.o.t..i.c.f.i..g.u..r.e.s.s,.l..i..g.h.t.m,.u.l.t..i.f.o.c..a.l....................................................................................... 1. Eosinophilicgranularityc,ytoplasmic,increased,moderate,diffuse 2. EMH, diffuse,moderately-severe .....................................M.@.9.t...i.@.....f.igu.r.es.,..m.i.n.i.ma.l.,....m...u...l...t...i.f.o...c...al.................................................. 19577-4 1. Eosinophilicgranularityc,ytoplasmic,increased,moderate, diffuse 2. EMH, diffuse,moderately-severe .....................................n.@.9.t...i.c..f.i.g.u.r.e.s.,..m.i.n.i.m.a.l......m..u...l...t...i...f.o...c..a...1................................................. 19579-3 1. Eosinophilicgranularityc,ytoplasmic,increased.moderate,diffuse 2. EMH, diffusem,oderately-severe Page 2 of 3 Incidenceand SeverityofLightMicroscopicLiverFindings MicroscopicFinding (numberexamined) Eosinophilicgranularityc,ytoplasmic, increased minimal slight moderate Cytoplasmic clearing minimal slight moderate Extramedullaryhematopoiesis moderate moderately-severe Mitotic figures,increased minimal slightI PFOS Dose Levels(mg/kg/day) 0 0.4 1.6 3.2 (5) (5) (5) (5) 2 2 1 - - 3 3 1 - - 1 4 - 1 2 - - 4 1 5 - 1 5 4 1 - - 1 4 5 2 1 2 2 -I - 2 Page 3 of 3 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.:418-013E,M-99.39 Animal No.: 19504-3 TreatmentGroup: 0 mg/kg,Control Sex:-non apj2licable Species/StrCrali:nCD:SBR VAF/Plus@ Rat Tissue: Liver BlockNo(s).: 99.39-1 Z15778 - Photo/NegativNeo(s).:Z15782 SignificanLtesions(checkone): Yes x No InterpretinPgathologis(tsignaturaend date): 3-10-99 Features: Z15778. Liver.Hepatocyteadjacentocentravleinatleftand bottom. Bile canaliculuasttop.Cytoplasmicorganelleasreclumped togetherwithinterveninsgpaces of finegranularmaterial.Mitochondriaappeardense and have some dilatedcristaeF.ive (5)peroxisomescounted. Z15779. Liver.HepatocyteexhibitinsgimilacrelluladretailassZ15778. Zero (0) peroxisomesclearlyidentified. Z15780. Liver.Hepatocyteappearssimilartoabove exampleswithclumped organelles. A hematopoieticellisinthetoprightsinusoidand a bilecanaliculuisspresentinthe lowerleft.Mitochondriaappeardenseand have some dilatedcristaeT.hree (3) peroxisomescounted. Z15781. Liver.Hepatocyteand adjacentsinusoidcontainingtwo nucleatedred blood cells.Mitochondriaappeardense withsome granulardepositsand dilatedcristae.Few numbers of peroxisomesidentifieadl,thoughpoor fixatiomnakes theirclearidentification difficulFto.ur (4)peroxisomescounted. Z15782. Liver. Centerhepatocyteissurroundedby severalotherhepatocytesw,itha sinusoidcontaininga nucleatederythrocytienbottom rightand a bilecanaliculuisntop portionof photograph.Mitochondriaappeardense withsome granulardepositsand dilatedcristaeF.our (4)peroxisomescounted. Conclusions: Normal hepatocyte.SuboptimalfixationA.verage 3.2peroxisomesper hepatocyte. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013;EM-99.39 Animal No.: 19504-3 TreatmentGroup: -0 mg/kg,control Sex:-non al2l2licable Species/StraCirnl::CD@BR VAF/Plus@ Rat Tissue: Lung BlockNo(s).: 99.39-2 Z15783 - Photo/NegativNeo(s).:_Z15787 SignificanLtesions(checkone): Yes Interi)retPiantghologis(tsignaturaend date): X No Features: Z 15783. Lung. Dark lining cellis a type II pneumocyte (PN2) which contains four lamellar bodies. Adjacent lining cellsare cuboidal withabundant granular material in their cytoplasm consistentwith glycogen. Subjacent to the surfaceisa capillaryand endothelialcell and a cellwith an oblong nucleus and abundant lipid-likmeaterialin itscytoplasm, probably a macrophage. Z 1584. Lung. Centraldark liningcellisa PN2 with severallamellarbodies. A largearea of granularmaterialin thiscellisprobably glycogen.Adjacent liningcellscontainabundant fine granularmaterial,probably glycogen as well. Dorsally isan isolatedcell,with numerous mitochondria and some endoplasmic reticulumand severaldense granules,probably a macrophage. A capillaryand endothelialcellisto the rightof the PN2. Z15785. Lung. A clusterof cuboidalepitheliac]ellswith abundant granularmaterialin their cytoplasm, apparentlyglycogen. This may be a terminalbronchioleor an alveolus whose lining cellshave not flattenedout.Subjacent to theseisa capillaryl,iningendothelialcelland a macrophage, whose cytoplasm containsabundant lipid-likderoplets. Z 15786. Lung. An alveoluswith severalcuboidal liningcells.All appear to be unflattened PN Is. The centralcellcontainsseveralvacuoles which may representempty lamellarbodies suggestiveof a PN2. The long cleftisprobably an artifactD.orsally isa capillaryand a couple endothelialcells. Z 15787. Lung. Area of collapsedalveoliand capillariesE.rythrocyteswithin capillaries. FlattenedPNis linethe alveolarspaces. No lamellarbodies evident. Conclusions: Normallung. Few type 11pneumocytes and lamellarbodies identified. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.:418-013E;M-99.39 Animal No.: 19505-1 TreatmentGroup:-0 ma/kg,control Sex:- non applicable Species/StrCarli:nC:D@BR VAF/PlusS Rat Tissue: Liver BlockNo(s).: 99.39-3 Z15788 - Photo/NegativNeo(s).:_Z15792 SignificanLtesions(checkone): Yes -X No InterpretinPgathologis(tsignaturaend date): Features: Z15788. Liver.Hepatocytecontainsabundantfinegranularmaterialand some clumping of mitochondriaand endoplasmicreticulum.Mitochondriaaredense and containsome dilatedcristaeF.ew peroxisomesarenoted,four(4)counted. Z15789. Liver.HepatocyteappearssimilartoZ15788, withfinegranularcytoplasmic materialand clumpingof largerorganellesF.our (4)peroxisomescounted. Z15790. Liver.Hepatocyte.Ventralliysa largesinusoidwitha Kupffercellamong severalerythrocytesA. hematopoieticellison thelowerright.Some dilatedblebsof rough endoplasmicreticulum(RER) areevidentinthecenterhepatocyte.. Mitochondria appeardense withsome granulardepositsand dilatedcristaeO.ne (1)peroxisome counted. Z15791. Liver.Hepatocyteappearpartialldyetachedfrom adjacentcellsand suboptimallyfixed.. Mitochondriaappeardense withsome granulardepositsand dilated cristaeT.wo (2)peroxisomescounted. Z15792. Liver.Hepatocyteappearsessentiallsyimilartothosedescribedabove. Abundant finegranularmaterialincytoplasm.. Mitochondriaappeardense withsome granulardepositsand dilatedcristaeS.ome dilatedblebsof RER noted.Four (4) peroxisomescounted. Conclusions: Normal hepatocyte.SuboptimalfixationA.verage 3.0peroxisomesper hepatocyte. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013E,M-99.39 Animal No.: 19505-1 Treatment Group: -0 mg/kg. Control Sex: - non applicable Species/Stracriin::CDOBR VAF/Plus@ Rat Tissue: Lung Block No(s).: 99.39-4 Z15793 - Photo/Negative No(s).: Z15797 SignificantLesions (check one): Yes InterpretingPathologist(signatureand date): X No Features: Z 15793. Lung. Alveolus with liningcells.Two cuboidal pneumocytes with finegranular cytoplasm. Top cellhas an extended flattenedportion thatcontains a lamellar-likestructure,and may be a PN2 thatisbecoming a PN 1. Normal, more flattenedPN Is are present adjacent to these two centralcells. Z15794. Lung. Two of four cuboidal liningcellshave lamellarbodies characteristiocf PN2. Smaller and darker cellshas a microvillous border than contains a lamellar structure,probably alveolarsurfactant. Z 15795. Lung. Alveolus with liningcells.Dark cuboidal cellisa PN2 and contains several lamellar bodies. Some flattenedPN 1s are present as well as another cuboidal pneumocyte. A capillarycontainingerythrocytesiswithin the alveolarseptum. Z 15796. Lung. Alveolus and area of collapsed septa. Two PN2s are present,both containing two to six lamellarbodies. No alveolarluminal lamellar structuresare evident. Z15797. Lung. Alveolus and liningcells.The dark columnar cellisa PN2 and contains several lamellarbodies. The leftadjacent pneumocytes appear not to have flattenedout as more differentiatedPNIS. A second PN2 isjustto the right of the centralPN2, and contains several small lamellarbodies. Subjacent tothe alveolus isa capillarycontaining a leucocyte and endothelialcell. Conclusions: Normal lung. A moderate number of type 11pneumocytes were identifiablea,nd contained lamellarbodies. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013E,M-99.39 Animal No.: 19506-4 TreatmentGroup: 0 mg/kg,control Sex: -non al2i)licable Species/Strain:Crl:CDOBR VAF/PlusS Rat Tissue: Liver BlockNo(s).: 99.39-5 Z15808 - Photo/NegativNeo(s).: Z15812 SignificanLtesions(checkone): Yes InterpretinPgathologist(signaturaend date): x No Features: Z15808. Liver. Hepatocytecontainsabundant finegranularcytoplasmic materialand some clumping of predominantlymitochondriaand rough endoplasniic reticulum.Five (5)peroxisomescounted. Z15809. Liver. HepatocyteappearssimilartoZ15808. Some dilatedblebsof RER are noted.Six (6)peroxisomescounted. Z15810. Liver. Hepatocytehas a washed-out appearancetomuch of itscytoplasm,and issurroundedby hematopoieticcellsventrallyE.ndothelialcellsareincompletely defined.At thetopleftof thephotographisa bilecanaliculusT.wo (2)peroxisomesare counted. Z1581 1.Liver. Hepatocytewith prominentbilecanaliculuastlower leftand sinusoidon rightmargin of photograph. Mitochondriaaredense and containsome dense depositand dilatedcristae.Six (6)peroxisomescounted. Z15812. Liver.Hepatocytecontainsabundant washed-outcytoplasm and clumping of RER and mitochondria.At thetop aretwo hematopoleticcellsand a bilecanaliculusis presentinlower rightofphotograph.Three (3)peroxisomescounted. Conclusions: Normal liver.SuboptimalfixationA.verage 4.4peroxisomesper hepatocyte TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013;EM-99.39 Animal No.: 19506-4 Treatment Group: -0 me/kg, control Sex:-non api)licable Species/StraCirnl::CD@BR VAF/Plus@ Rat Tissue: Lung Block No(s).: 99.39-6 Z15818 - Photo/NegativeNo(s).: Z15822 SignificantLesions (check one): Yes InterpretinPgathologist(signatureand date): x No Features: Z15818. Lung. Dark cellwith round nucleusinthecenterisa PN2 which partially linesthealveolarspace. Severallamellarbodiesarewithinitscytoplasm. The largeclear cytoplasm representsan adjacentPNI. A capillarcyontainingan erythrocytewithintheseptum isimmediatelyadjacentthePN2. The remainder ofthetissueiscollapsedalveoliand septa. Z 15819. Lung. A thin-marginof PN I surroundsa protrudingseptum and capillariynthe middle ofthe photograph. Two endothelialcellsappear tolinethecapillaryw,hich containsan erythrocyte.To therightisa somewhat darkerPN2, which containsseverallamellarbodies. In the centralbottom isa largetypeIpneumocyte, with a largepolygonal nucleusand mostly clear cytoplasm. Z]5820. Lung. PNIs lineseveralseptainthisphotograph. Dorsally,thesepta]capillary containsa lymphocyte and a nucleatederythrocyte.In thecentercapillarya granulocyticcellis evident.No discerniblePN2s or lamellarbodiesareevident. Z 1582 1. Lung. Alveolusand liningcells.A PN2 ispresentliningthealveolus.Itisthedarker cellwith severaldilatedlamellarbodies.Most ofthe lamellarmaterialinthesebodies is missing. Two PN Isareevidenton therightmargin of thisalveolarspace. Large nucleuson the leftappearsto belong toan endothelialcelllininga septalcapillary. Z 15822.Lung. Alveolus and severalliningcells.Most appeartobe PN Is with abundant somewhat clearcytoplasm,which hasn'tcompletelyflattenedout. No definitivPeN2s are evident,althoughtwo small vacuolesina liningpneumocyte atthebottom of thephotograph may representlamellarbodies. A capillarycontainingtwo nucleatedcellsoccupiestherightside of theph tograph. Conclusions: Normal lung. Occasional type 11pneumocytes containing lamellarbodies identifiedN.o lamellarmaterialnoted withinalveolarspaces. TRANSNUSSION ELECTRON NHCROGRAPH INTERPRETATION StudyNo.: 418-013E,M-99.39 Animal No.: 19509-1 Treatment Group: 0 me/kg. control Sex: -non apl)licable Species/StraCriln::CDOBR VAF/Plus@ Rat Tissue: Liver Block No(s).: 99.39-7 Z15823 - Photo/Negative No(s).: Z15827 SignificantLesions (check one): Yes InterpretingPathologist(signatureand date): x No Features: Z15823. Liver. Hepatocyte bordered by sinusoidcontaining nucleated erythrocyte on leftand other hepatocytes on bottom and right.An endothelialcellisin the upper leftportion of the photograph. There isclumping of the mitochondria and RER with an interveningopen area or areas filledwith fine granular material.The RER is dilated. Mitochondria contain some granular deposits. Five (5)peroxisomes counted. Z15824. Liver. Hepatocyte. At the top isa nucleated erythrocyte.Sinusoidal endothelium ispoorly delineated.The RER shows some blebbing and dilatation,and mitochondria have some dense deposits and some dilatationof cristae.Five (5) peroxisomes counted. Z15825. Liver. Hepatocyte surrounded by hematopoietic cells.The RER and nuclear envelope shows some dilatationand blebbing. Mitochondria appear dense and have some granular deposits and dilatedcristae.Seven (7) peroxisomes counted. Z15826. Liver. Hepatocyte appears essentiallysimilarto above described cellswith clumping of major organelles. Five (5)peroxisomes counted. Z 15827.. Liver. Hepatocyte surrounded by hematopoietic cells.A singlelargervacuole is present in the cytoplasm and contains a flocculentlikematerial.Four (4) peroxisomes counted. Conclusions: Normal hepatocyte. Suboptimal fixation.Average 5.2 peroxisomes per hepatocyte. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013,EM-99.39 Animal No.: 19509-1 Treatment Group: -0 mg/kg, control Sex: -non al2j2licable Species/StraCirnl::CDOBRVAF/Plus@ Rat Tissue: Luniz Block No(s).: 99.39-8 Z15828 - Photo/NegativeNo(s).:_Zl5832 SignificantLesions(check one): Yes InterpretinPgathologist(signatureand date): x No Features: Z15828. Lung. Alveolusand liningcells.The darkercuboidalcellisa PN2 and containsseverallamellarbodies.An erythrocyteisimmediatelytotheleftof thePN2 and appearstobe withinthealveolus.At top leftisa pneumocyte which has two smalllamellar-like bodiesand isprobablya PN2 intransitiotnoa PN 1. A flattenedPN I isimmediatelyon top and totherightofthedark PN2. Beneath thesecellstheseptaarecollapsedtogether. Z15829. Lung. Alveolusand liningcells.The smallerdark cellisa PN2 with severallamellar bodies.To itsrightisaflattenePdNI and to itsleftisacuboidalPNI. Thelamellarormyelinlikematerialtothe rightofthePN2 appearstobe withina capillarlyumen. At thebottom right isa thin-walledseptum containinga capillaraynd erythrocyte. Z 15830. Lung. Two erythrocytesarewithinthecapillarylumen. The liningpneumocytes are flattenedand appear tocontainseveralvacuolesthatmay have containedlamellarbodies. Subjacentarecollapsedseptaand a mixture of septalcellsincludingpneumocytes and endothelialcells. Z 1583 1. Lung. Area of collapsedalveolarsepta.At theleftmiddle and rightmiddle-topare dark cellswhich containlameliarbodies and arePN2s. Other cellsarecompressed pneumocytes, endothelialcells,and blood cellswithincapillariesS.ome lamellardebrisappearsto be within capillarylumina. Z 15832. Lung. Area ofcollapsedsepta.The largeround clearcellisa cuboidalpneumocyte. Dorsalto thisareendotheliaclellslininga capillaryT.he lamellarstructuraeppearstobe within the lumen of thecapillary. Conclusions: Normal lung. Scattered type 11pneumocytes present. Some lamellar materialalsopresentwithincapillariesN.o lamellarbodies noted in alveolarspaces. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.:418-013E,M-99.39 Animal No.: 19516-2 TreatmentGroup:-0 mg/kg.control Sex: non applicable Species/StrCrali:nCD:OBR VAF/Plus@ Rat Tissue: Lung BlockNo(s).: 99.39-10 Z15838 - Photo/NegativeNo(s).Z:15842 SignificaLnetsion(scheckone):_ Yes InterpretinPgathologis(tsignaturaend date): x No Features: Z15838. Lung. Two pneumocytesatthebottomarePN2 and containsome lamellarbodies.Dorsaltothem arecollapsedseptacontainingprimarilypneumocytes and endotheliaclells. Z15839. Lung. Severalpneumocytes arepresent.Centralpneumocyte containsseveral lamellarbodiesand isa PN2. FlattenedPNIs linetheprotrudingseptum which contains a capillarye,ndotheliaclelland leucocytewithinthecapillary. Z15840, Lung. Singleseptumlinedby PNIs on eitherside.On therighttopisa PN2 with severallamellarbodies.The rightlowerpneumocyte alsocontainsa singlelamellar body. The septalcapillaraynd endotheliumarealsoevident. Z15841. Lung. Area ofcollapsedsepta.At rightaretwo pneumocyteswhich contain multiplelamellarbodies.Other compressed septa]cellsappeartobe pneumocytes and endotheliaclells. Z15842. Lung. Area of collapsedsepta.Severalcuboidalpneumocytesarepresent. Centrallyisa capillarcyontainingan erythrocyteN.ext toitisa couplelamellar structuretshatappeartobe withinthecapillarlyumen. Conclusions: Normal lung. Random typeIIpneumocytes containinglamellarbodies were identifiedN.o lamellarbodieswere clearlyidentifiewdithinalveolarspaces. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013E,M-99.39 Animal No.: 19516-2 Treatment Group: -0 miz/kg,control Sex: - non Lapi)licable Species/StraCirni::CDOBVRAF/PlusS Rat Tissue: Liver Block No(s).: 99.39-9 Z15833 - Photo/NegativeNo(s).: Z15837 SignificantLesions (check one): Yes -X No InterpretingPathologist(signatureand date): Features: Z15833 Liver. Hepatocyte with a sinusoidat lower rightand a bile canaliculuson the left.Cell has abundant washed-out cytoplasm and clumping of mitochondria and RER. There isdilatationof RER and the nuclear envelope. Mitochondria contain some granular deposits and some dilatedcristae.Plasma membranes and sinusoidalendothelialcellsare poorly delineated. Five (5)peroxisomes counted. Z15834. Liver. Dilationand blebbing of RER. Some granular deposits present in mitochondria. Nine (9)peroxisomes counted. Z 15835. Liver. Mitochondria contain some dense deposits and dilatedcristae.There is blebbing of the RER. Plasma membranes are poorly defined. Five (5)peroxisomes counted. Z15836. Liver. Hepatocellularmorphology essentiallysimilaras those described above. Four (4) peroxisomes counted. Z15837. Liver. Mitochondria contain some dense deposits and dilatedcristae.Plasma membranes are poorly defined. A multivesicularbody ispresent to the rightof the nucleus. Cell at lower leftappears to be an endothelialcell,but may be a hematopoietic cellin the sinusoids. Two (2) peroxisomes counted. Conclusions: hepatocyte. Normal liver.Suboptimal fixation.Average 5.0 peroxisomes per TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.:-418-01E3M-99.39 Animal No.: 19534-1 TreatmentGroup: 0.4m-g/kg Sex:-non al2RIicable Species/StrCrali:nCD:OBR VAF/Plus@ Rat Tissue: Liver BlockNo(s).: 99.39-31 Z15935 - Photo/NegativNeo(s).:--ZI5939 SignificanLtesions(checkone): X Yes No InterpretinPgathologis(tsignaturaend date): Features:Z15935. Liver.Hepatocytewithpoorly-definepdlasmamembrane and no surroundingendotheliaclells.Some hematopoieticellsarearound thishepatocyte. Mitochondriacontainsome granulardepositsand some dilatecdristae.RER isdilated and shows some blebbing.Four (4)peroxisomescounted. Z15936. Liver. Hepatocyteshows marked dilatioonf RER and poor delineatioonf the plasma membrane. The nuclearenvelopeisalsodilated.A largeclearvacuole-ipsresent intherightsideof thecell.Eighteen(18)peroxisomescounted. Z15937. Liver.Hepatocytewithclumpingof organelleasnd interveninwgashed-out areasof cytoplasm.Mitochondriacontainsome granulardepositsand some dilated cristae.RER isdilatedand shows some blebbing.Eleven(11)Peroxisomescounted. Z15938. Liver.Hepatocyteshows similarultrastructumroarlphology as thosedescribed above,with suboptimalfixationA. multivesiculabrody ispresenttotherightof the nucleus.Eleven (11)peroxisomescounted. Z15939. Liver.Hepatocytewithclumping oforganelleasnd interveninwgashed-out areasof cytoplasm.Plasma membrane isindistinctA.hematopoieticellisatthebottom ofthephotograph.Mitochondriacontainsome granulardepositsand some dilated cristaeR.ER isdilatedand shows some blebbing.Sixteen(16)peroxisomescounted. Conclusions:Thereappearstobe increasednumbers ofperoxisomesperhepatocyte compared withcontroliver.Average 12.0peroxisomesperhepatocyte.Cellularfixation suboptimal. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013EM-99.39 Animal No.: 19534-1 Treatment Group: 0.4 m@/kg Sex:-non applicable Species/StraiCnrl::CDOBR VAF/PlusS Rat Tissue: Lung Block No(s).:-99.39-32 Z15941 - Photo/NegativeNo(s).: 15945 SignificantLesions (check one): Yes -X No InterpretinPgathologist(signatureand date): Features: Z15941. Lung. Alveolusand liningcells.Alltheliningcellsappeartobe typeI pneumocytes (PNIs). Beneath thePNIs isa round topolygonalcellcontaining5 lamellarbodies initscytoplasm;thisisa type 11pneumocyte (PN2). This isprobablya liningcell,and theplane of sectionissomewhat flatthrough theseptalwall. ZI 5942. Lung. Alveolarspace and liningcells.The dark and somewhat flattenedliningcellisa PN2 and containsseveralvacuolesempty of contents.These appear tobe "empty" lamellar bodies.The lighter-stainipnngeumocyte atthefarleftalsocontainstwo similar-appearing vacuoles. Z15943. Lung. Alveolusand liningcells.The dark nucleusispartof a flattenepdneumocyte, but alsoattacheswithtwo adjacentmore lucentPNIS. No lamellarbodiesnoted inthis photograph,althougha vacuoleadjacenttothetop leftnucleusmay be an "empty" lamellar body. Septalwallsand alveoliarecollapsedand normal pulmonary architecturiesindistinct. Z15944. Lung. The dark liningcellon therightcontainsseverallamellarbodiesand isa PN2. The otherliningcells,althoughcuboidal,appear tobe pneumocytes which have not flattenedout inthecharacteristitcype Ipneumocyte fashion. ZI 5945. Lung. Alveolusand liningcells.The two dark liningcellsboth appearflatteneadlong the septalwall and arePNIS. No lamellarbodiesareseen intheseliningcells.The topcellin the photograph appearsdetached and surroundedby some extracellulalripid-likmeaterial.Its identityisunclear,but may be a macrophage orendothelialcell.A capillaryispresentatthetop left. Conclusions: Essentiallynormal fetallung. Random type 11pneumocytes with lamellar bodies identified.No remarkable abnormalitiesnoted. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-01E3M-99.39 Animal No.: 19536-3 TreatmentGroup: -0.4me/k2 Sex:-non a]2plicable Species/StrCrali:nCD:@BR VAF/PlusS Rat Tissue: Liver BlockNo(s).: 99.39-33 Z15946 - Photo/NegativNeo(s).:-Z15950 SignificanLtesions(checkone): X Yes No InterpretinPgathologis(tsignaturaend date): Features:Z15946. Liver.Hepatocyte.ThereisdilatatiofnRER and ofthenuclear envelope.Some granulardepositsareevidentwithinsome mitochondria.Four (4) peroxisomes counted. Z15947. Liver.Hepatocyte.Thereissome clumpingof organellesT.he RER and nuclearenvelopearedilated.Seventeen(17)peroxisomescounted. Z15948. Liver.Hepatocyte.Thereispoordefinitioonf theplasmamembrane. RER and thenuclearenvelopearedilatedF.ourteen(14)peroxisomescounted. Z15949. Liver.Hepatocyteappearssimilartothosedescribedabove.Eighteen(18) peroxisomes counted. Z15950. Liver.Hepatocyte.At thetopisan endotheliaclelland severaladjacent erythrocyteasnd one nucleatederythrocyteT.he plasma membrane of thehepatocyteis indistinacntd organelleasreclumped inthecytoplasm.RER isdilatedE.ight(8) peroxisomes counted. Conclusions: Increasednumber ofperoxisomesperhepatocytecompared withcontrol samples. Average 12.2peroxisomesperhepatocyte.Suboptimalfixation. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-013EM-99.39 Animal No.: 19536-3 Treatment Group: 0.4 ma/k2 Sex: -non gpl2licable Species/StraiCnr:l:CDOBRVAF/Plus@ Rat Tissue: Luniz Block No(s).: 99.39-34 Z15951 - Photo/Negative No(s).: Z15955 SignificantLesions (check one): Yes InterpretingPathologist(signatureand date): X -No Features: Z15951. Lung. Alveolus and areaof collapsedsepta.The bottom alveoluscontains debrisincludingsome lamellarmaterialconsistentwith secretedlamellarbodies (surfactant) from PN2s. The liningpneumocytes are allflattenedand appear to be PN Is. No PN2s are identified.At leasttwo capillarieasre present in thecollapsed septalarea. Z]5952. Lung. Alveolus and liningcells.PNIs linethe alveolarspace and are flattened.On the rightthe nucleus and cellbeneath the PN I containsa singledark lamellarbody, which suggest thiscellmaybe a PN2. Other cellsin the photograph are endothelialcells,granulocytein a capillaryand some nonidentifiedcells.No lamellarbodies are present in the alveolarlumen. Z 15953. Lung. The prominent dark lamellaror myelin-likebody inthe centerof the photograph iswithin a capillaryspace. Type I pneumocytes are flattenedand linethe alveolarspace. At the bottom of the photograph in thecollapsedseptalarea is a cellwhich containstwo lamellar bodies,a PN2. No abnormalitiesnoted. Z15954. Lung. Alveolus and liningcells.A small centraldark cellcontains3-4 lamellarbodies and isa PN2. The adjacentcelliscuboidal,but appears to be a pneumocyte which has not flattenedout. A capillarycontainingseveralerythrocytesand a liningendothelialcellis prominent in thecenterof the photograph. Some debrisispresentin the alveolarlumen, and some may be finelamellarmaterial.The nucleatedcellat the top containstwo lamellarbodies and isalso a PN2. Z15955. Lung. Lamellar bodies withina PN2 linethe alveolarspace. Collapsed alveoliand septafillthe top of the page. No abnormalitiesnoted. Conclusions: Essentiallynormal fetallung. Some type 11 pneumocytes identified containing lamellar bodies. Some lamellar bodies presentwithin alveoli. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.:-418-01E3M-99.39 Animal No.: 19543-4 TreatmentGroup: -0.4 mg/kg Sex:-non ai)i)licable Species/StrCrali:nCD:OBR VAF/Plus@ Rat Tissue: Liver BlockNo(s).: 99.39-35 Z15956 - Photo/NegativNeo(s).:-Zl5960 SignificanLtesions(checkone): x Yes No InterpretinPgathologis(tsignaturaend date): Features:Z15956. Liver.HepatocytewithclumpingofRER and n-iitochondria. Plasma membranes areindistinctC.ross-sectionosf RER appeardark. A bilecanaliculus ispresentatthe topright.Severalhematopoietic ellsarearound the periphery. Mitochondriacontainsome dilatecdristaeE.ight(8)peroxisomescounted. Z15957. Liver. HepatocytemorphologyappearssimilatroZ15956. Organellesare clumped in thecytoplasm;interveninsgpacescontainfinegranularmaterialwhich is probablyglycogenand cytosolidcebris.Five(5)peroxisomescounted. Z15958. Liver. Hepatocytewithbilecanaliculpiresentattopand bottomofcell. Fifteen(15)peroxisomescounted.Fixationisverypoor. Z15959. Liver.Hepatocytewithmarked clumping oforganellesT.wo hematopoietic cellsarepresentatthebottom of thephotograph.Three (3)peroxisomescounted. Z15960. Liver.Hepatocytesurroundedprimarilyby hematopoleticells.Sinusoidal endotheliaclellsareabsentdue topoor fixationH.epatocyticmitochondriacontainsome granulardepositsand dilatedcristaeN.ine (9)peroxisomescounted. Conclusions: Sectionofliverexhibitsuboptimaltopoorfixation.Numbers of peroxisomeperhepatocyteonlyveryslightlhyigherthancontrolvalues.Average 8.0 peroxisomes perhepatocyte. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013 EM-99.39 Animal No.: 19543-4 Treatment Group: 0.4 mg/kg Sex: -non applicable Species/StraiCnr:l:CDOBRVAF/Plus@ Rat Tissue: Lung Block No(s).: 99.39-36 Z15961 - Photo/Negative No(s).: Z15965 SignificantLesions (check one): Yes x No InterpretingPathologist(signatureand date): Features: Z 1596 1. Lung. Alveolus and liningcells.Lining cellsappear cuboidal,and may be a terminalbronchiole. The centralcuboidalcellcontainsseverallamellarbodies within a fine granularcytoplasmic background. On eithersidearetwo darker compressed cellswhich contain apicalvacuoles. Some lamellarmaterialispresent in the alveolarspace. Z15962. Lung. Severalcuboidalpneumocytes linethisairway or alveolarspace. They all containabundant granularmaterialin theircytoplasm,which isconsistentwith glycogen. No PN2s are evident. A flattenedPN I ispresentat the righttop of the alveolarspace. Two septal capillarieasrepresent,one dorsallyand one ventrally.Both contain some lamellarmyelin-like materialin theirlumina. Z15963. Lung. Photograph shows severalcuboidal pneumocytes, the top cellof which contains a clearvacuole suggestiveof an "empty" lamellarbody. A dark erythrocyteispresent in the capillaryto the leftof the liningcells. The cellat the bottom leftcontainsa few small lamellar bodies and isprobably a PN2. No lamellarbodies noted inalveoli,and no abnormalitiesnoted. Z 15964..Lung. Alveolus and liningcells.The liningcellsarepredominantly flattenedand appear to be differentiatePdN Is. Several mitochondria and a dark vacuole are present in the centralliningcell.The dark vacuole may representa depletedlamellarbody. The dark lamellar myelin-likestructurenear the top iswithina capillaryspace with a collapsed septum. Z 15965.,Lung. The cellatthe bottom of the photograph containsseverallamellarbodies and is a PN2 within the collapsedseptum. PN]s linethe airspace on the fight.Two capillaries containingdark erythrocytesare beneath the surfaceepithelium. No abnormalitiesare noted. Conclusions: Essentiallynormal fetallung. Several type Ilpneumocytes identifiedwith lamellar bodies. Also, some lamellarstructuresnoted within alveolarspaces. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013EM-99.39 Animal No.: 19546-1 Treatment Group: 0.4 m2/kiz Sex: -non applicable Species/StraCriln::CDOBR VAF/Plus@ Rat Tissue: Liver Block No(s).: 99.39-37 Z15966 - Photo/NegativeNo(s).: Z15970 SignificantLesions (check one): X Yes No InterpretingPathologist(signatureand date): Features: Z 15966. Liver. Hepatocyte. Plasma membrane isindistinctand cytoplasmic organelles are clumped together. Mitochondria show some dense deposits and dilated cristae.Fifteen(15) peroxisomes counted. Z15967. Liver. Hepatocyte appears morphologically similarto Z15966, with clumping of organelles,indistinctplasma membranes, and mitochondrial changes. RER appears dilated.Seven (7) peroxisomes counted. Z15968. Liver. Hepatocyte. A prominent bilecanaliculusispresentin the bottom right corner. Organelles are clumped with large interveningspaces of fine granular material. Mitochondria show some dense deposits and dilatedcristae.Twenty (20) peroxisomes counted. Z 15969., Liver. Hepatocyte appears essentiallysimilarto Z 15968. Fixation ispoor. Eleven (11) peroxisomes counted. Z15970. Liver. RER isdilatedand plasma membranes are indistinct.An endothelialcell is not present at sinus at top. Mitochondria show some dense deposits and dilatedcristae. Twelve (12) peroxisomes counted. Conclusions: Liver exhibitspoor fixation.There appears to be increased numbers of peroxisomes per hepatocyte. Average 13.0 peroxisomes per hepatocyte. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-01E3M-99.39 Animal No.: 19546-1 TreatmentGroup:-0.4 mg/kq Sex:-non applicable Species/StrCrali:nCD:OBR VAF/Plus@ Rat Tissue: Lung BlockNo(s).: 99.39-38 Z15971 - Photo/NegativNeo(s).: Z15975 SignificanLtesions(checkone): Yes InterpretinPgathologis(tsignaturaend date): x No Features: Z15971. Lung. Alveolusand liningcells.Severalliningcellscontain lamellarbodies,most predominantlythecentercellwithatleastninelamellarbodies. Thisisa distinctivPeN2. The threecellsatthetopof thephotographappeartobe pneumocytes whose alveoliareoutof planeof sectionor arecollapsed. Z15972. Lung. Photographslightloyutof focus.An unattachedPN2 ispresentinthe alveolarspace.Both sidesoftheseptum arelinedby flattenePdNIS. A capillaraynd endotheliaclellareevidentwithintheseptum. No lamellarmaterialispresentin the alveoli. Z15973. Lung. Alveoliand septum.The darkcellon thetopleftisa PN2 withseveral "empty" lamellarbodies.The otherliningcellsarePN Is. A capillarwyitha lun-tinal erythrocytiespresentinthecenterof theseptum.No lamellarmaterialispresentinthe alveoli. Z15974. Lung. Alveolusand liningcells.A flattenePdN I linesthebottom marginof thealveolus.Next toitisa cuboidalpneumocyte withseveralclearvacuoles,a PN2. The vacuolesare"empty" lamellarbodies.The centralcellwitha microvilloubsorder, dilatednuclearenvelopeand distendedendoplasmicreticulumisa cuboidalpneumocyte exhibitinsgome elementsof suboptimalfixationN.o lamellarbodiesnotedinthealveoli. Z15975. Lung. Cuboidalliningcellincenterrightand cellatcenterleftboth containone ortwo vacuoleswhich appeartobe lamellarbodies.These cellsareprobablyPN2s convertingtoPNis afterexpressintgheirlamella(rsurfacta)nmtaterial.Nolamellar materialnotedinthealveoli. Conclusions: Essentiallnyormal fetalung. ScatteretdypeIIpneumocytes withlamellar bodiesidentifiedN.o lamellarmaterialispresentinthealveoli. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013EM-99.39 Animal No.: 19546-2 Treatment Group: 0.4 mg/kg Sex: -non aprlicable Species/StraCirnl:CDOBRVAF/Plus@ Rat Tissue: Liver Block No(s).: 99.39-39 Z15976 - Photo/Negative No(s).:__ZI5980 SignificantLesions (check one): Yes -X No InterpretingPathologist(signatureand date): Features: Z15976. Liver. Hepatocyte exhibitsa poorly-definedplasma membrane. The organellesare clumped and the mitochondria show dilatedcristae.At top rightisan interstiticaelllsurrounded by collagen,probably a fibroblast.At the lower leftare some hematopoietic cellswithin a sinusoid. Four (4)peroxisomes counted. Z15977. Liver. Two hepatocytes with an interveningbilecanaliculus.Hematopoietic cellsare present in the upper rightand lower leftsinusoids. The largercytoplasmic organelles are clumped togetherand the cytosol appears washed-out or contains a fine granular material. The mitochondria show some dilatedcristae.The hepatocyte (bottom cell)contains seven (7) peroxisomes. Z15978. Liver. Central hepatocyte in photograph appears similarto those described above, fixationispoor. Two (2)peroxisomes counted. Z15979. Liver. Hepatocyte, whose morphology issimilarto those described above. Five (5) peroxisomes counted. Z15980. Liver. Hepatocyte exhibitsa poorly-defined plasma membrane. The organelles are clumped and the mitochondria show dilatedcristae.RER isdilated.Nine (9) peroxisomes counted. Conclusions: Essentiallynormal liver.Poor cellularfixation.Average 5.4 peroxisomes per hepatocyte. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013EM-99.39 Animal No.: 19546-2 Treatment Group: 0.4 mg/kg Sex: -non applicable Species/StraCirnl:CDOBRVAF/PlusS Rat Tissue: Lung Block No(s).: 99.3940 Z15981 - Photo/Negative No(s).:--Z 15985 SignificantLesions (check one): Yes InterpretingPathologist(signatureand date): X No Features: Z15981. Lung. Alveolus and liningcells. A partiallydetached pneumocyte ispresent in the airspace. Large rightcentralcellcontains a singlelamellar body near its top margin. The other dark organellesappear to be predominantly n-iitochondria.The cytoplasm of these cuboidal pneumocytes contains a finegranular material suggestive of glycogen. Z15982. Lung. Central cellcontains multiple lamellarbodies and isthe cytoplasm of a PN2. This is in an area of collapsedalveoliand septa. A capillaryand endothelialcell are atthe leftof the photograph. The two dark cellson eitherside of the PN2 are not clearlyidentifiableT.he bottom rightcellisa PNI. No abnormalitiesdescribed. Z15983. Lung. Two columnar cellson the rightboth contain lamellarbodies, although the majority of theircytoplasm consistof finegranular material. PN Is appear to be atthe bottom of the photograph and a capillaryand endothelialcellare atthe left. Z15984. Lung. An area of collapsed alveoliand septa. At the top and bottom are two round to polygonal cellsthatcontain some lamellarbodies consistentwith type Il pneumocytes (PN2s). Some lamellar materialappears to be extracellularadjacent to the bottom PN2. The centralcellsallappear to be PNIS. At the bottom rightisa capillary, endothelialcelland erythrocyte.No lamellarbodies noted in alveoli. Z 15985., Lung. At the top rightisa cuboidal PN2 liningthe alveolus. This cellcontains severallamellar bodies,and sitsnext to a more flattenedPN 1. Centrally isa capillary containing two largeendothelialcellnuclei and an erythrocyte. Conclusions: Essentiallynormal fetallung. Random type IIpneumocytes (PN2s) containing lamellar bodies were identified.Some extracellularlamellarmaterial identifiedoutside one type Ilpneumocyte. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013EM-99.39 Animal No.: 19560-1 Treatment Group: -1.6m-g/kg Sex: -non api)licable Species/StraCirnl::CDOBVRAF/Plus@ Rat Tissue: Liver Block No(s).: - 99.39-21 Z15884 - Photo/Negative No(s).:-Z15888 SignificantLesions (check one): x Yes No InterpretingPathologist(signatureand date): Features: Z15884 Liver. Hepatocyte. Plasma membranes and adjacentendothelial cellsare indistinctc,onsistentwith suboptimal fixation.Dilationof RER with some blebs. Some dilatedcristaein mitochondria. A singleclearvacuole ispresent in the cytoplasm on the rightside of the cell.Fifteen(15) peroxisomes counted. Z15885. Liver. Some granular depositspresent in mitochondria and dilatationof RER. Twenty-one (21)peroxisomes counted. Z15886. Liver. Hepatocyte in center of photograph with hematopoietic cellsat top. Some clumping of mitochondria and RER. Dilated RER. Six (6)peroxisomes counted. Z15887. Liver. Hepatocyte containing a singlelarge lipiddroplet indorsal cytoplasm. At top rightis a hematopoietic celland atlower leftisa bilecanaliculus.There is clumping of cytoplasmic organelles.RER and nuclear envelope appear dilated. Five (5) peroxisomes counted. Z15888. Liver. Hepatocyte similarto those described above. Some stainprecipitateis present on photograph. Eleven (11) peroxisomes counted. Conclusions: There appears to be increased numbers of peroxisomes per hepatocyte compared with controlsamples. Average 11.6 peroxisomes per hepatocyte. Suboptimal tissuefixation. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013 EM-99.39 Animal No.: 19560-1 Treatment Group: 1.6 mg/kg Sex: --non applicable Species/StraiCnr:l:CD@)BR VAF/Plus@ Rat Tissue: Lung Block No(s).: 99.39-22 Z15889 - Photo/Negative No(s).: Z15893 SignificantLesions (check one): Yes -X No InterpretingPathologist(signatureand date): Features: Z15889. Lung. Alveolus and liningcells.Top cellwith darker cytoplasm isa PN2 and containstwo lamellarbodies. The lower cellalsocontainstwo prominent vacuoles,which are probably expressed lamellarbodies even though the lamellarmaterialisno longer evident. The centralcellisa PN 1. Beneath the PN I isan extension of an unidentifiedcellout of plane of section.A capillaryispresentat the bottom leftof the photograph. No lamellarmaterialis present in the alveolarspace. Z 15890. Lung. Three liningcellsand a capillaryare evident.The centraldark cellwith an oblong nucleus isa PN2, and itcontains3 discerniblelamellarbodies. At the top leftand to the rightof the PN2 are PN Is with more lucentcytoplasm. The capillaryand liningendothelialcell are atthe top right.A dark erythrocyteispresentin the capillary. Z15891. Lung. Alveolus, liningcells,and capillariesC.entrallyisa PN2 somewhat sandwiched in between two flattenedPNIs liningthe alveolus. The PN2 contains several lamellarbodies withinitscytoplasm. Capillariecsontainingdark erythrocytesand linedby endothelium are presentat the upper leftand lower rightof the photograph. No lamellarmaterial ispresent in the alveolarspace. Z]5892. Lung. Featuresare similarto those describedforZ15891. A singlePN2 ispresent between two PN Is liningthe alveolus.The PN2 containsseverallamellarbodies.The cell beneath the PN2 has an irregularnucleus,some prominent RER and issurrounded by some collagen;thisappears to be a fibroblast. Z15893. Lung. Alveolus with severalcuboidalpneumocytes. There are not clearlydistinctive PN2s inthisphotograph. The capillaryat the bottom containsa lamellarmyelin-linestructure within itslumen. No lamellarmaterialispresentin the alveolarspace. Conclusions: Essentially normal lung. Several type II pneumocytes with lamellar bodies were identified.Fixation was considered acceptable. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.:-418-01E3M-99.39 Animal No.: 19560-2 TreatmentGroup:- 1.6miz/kiz Sex:-non apl)licable Species/StrainC:rl:CD@BR VAF/Plus(O) Rat Tissue: Liver BlockNo(s).-:00.39-23 Z15894 - Photo/NegativNeo(s).: Z15898 SignificanLtesions(checkone): x Yes No InterpretinPgathologis(tsignaturaend date): Features:Z15894. Liver.Hepatocytewithsurroundinhgematopoieticells.Thiscell appearspartialldyetachedfrom adjacenthepatocytesa,lthoughtwo hepatocytesare presentinthelowerrightofphotograph,allthreesharinga common bilecanaliculus. There isclumping of mitochondriaR,ER and some peroxisomes.Some dilatatioonf cristaiesnoted.Thirteen(13)peroxisomescounted. Z15895. Liver.HepatocyteappearssiniilarcteolldescribedinZ15894 withclumpingof organellesS.ome granulardepositsnotedin ntitochondriaF.ourteen(14)peroxisomes counted. Z15896, Liver.Hepatocytehasclumpingof organellewsithintervenincgytoplasmfilled withfinegranularmaterialp,robablypredominantlyglycogen.Mitochondriaappear denseand containsome granulardepositsand dilatedcristaeT.en (10)peroxisomes counted. Z15897..Liver.Hepatocyteappearssimilarto thosedescribedabove. Plasma membranes areindistinctA.hematopoieticellisatthebottom of thephotograph. Fourteen(14)peroxisomesarecounted. Z 15898.,Liver.Hepatocytemorphology essentiallsyame as above. A bilecanaliculus servesfourattachedhepatocytesinthetopcenterofthephotograph.Thir-tee(n13) peroxisomes counted. Conclusions: There appeartobe increasednumbers of peroxisomesperhepatocyte. Average 12.8per hepatocyte.Suboptimalfixatioonf tissue. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.:-418-013EM-99.39 Animal No.: 19560-2 Treatment Group: ---1.6mg/kiz Sex: non al2plicable Species/StraCirnl:CDOBRVAF/Plus@ Rat Tissue: Lung BlockNo(s).: 99.39-24 Z15899 - Photo/NegativNeo(s).: Z15903 SignificanLtesions(checkone): x Yes No InterpretinPgathologis(tsignaturaend date): Features: Z15899. Lung. Alveolusand liningcells.Centralcellisa PN2 with several basilarlamellarbodies.Adjacent liningcellsappeartobe PNIS. A freeerythrocyteis presentinthealveolarspace. At theleftofthe photographisa smallcapillarycontaining a lun-@nalerythrocyte.No abnormalitiesnoted. Z 15900. Lung. Alveolus and liningcells.At topcenterisa singlePN2 which contains fivelamellarbodies.Two lamellarmyelin-likestructureisnmiddle leftof photographare withincapillarlyumina. No lamellarmaterialisidentifiewdithinalveolarspaces. Z15901. Lung. Alveolus and liningcells.Nucleatedcellfreeinalveolusappearstobe a lymphocyte. Two centerliningcellsbothcontainlamellarbodiesand are PN2s even though one is"light"and one is"dark".Large cuboidalpneumocyte atlower left containssome lamellarbodies and connectstothe nextflattenedPN I segment. Z15902. Lung. A clusterofpneumocytes, fourof which containlamellarbodies characteristoifc PN2s. No famellarmaterialisidentifiewdithinalveolarspaces. Z15903. Lung. A group ofcuboidaland flattenepdneumocytes and ventralcapillary withina septalwall.Two cellscontaincleftsb,ut none containdefinitivleamellarbodies. No abnormalitiesnoted. Conclusions: Type Ilpneumocytes fairlyeasilyidentifieidnthisanimal,and atleast one clusterof PN2, possiblysuggestiveof increasednumbers of PN2. Otherwise ultrastructuramlorphology appearedessentiallsyimilartocontrolsections. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.:418-01E3M-99.39 Animal No.: 19560-3 TreatmentGroup: - 1.6mg/kg Sex: on aj2i)licable Species/Strcra]i:nCD:OBR VAF/Plus@ Rat Tissue: Liver BlockNo(s).: 99.39-25 Z15904 - Photo/NegativNeo(s).: Z15908 SignificanLtesions(checkone): x Yes No InterpretinPgathologis(tsignaturaend date): Features:Z15904. Liver.Hepatocyteborderedby hepatocyteson threesidesand sinusoidwithhematopoieticellon right.Bilecanaliculaittopand on left.There is clumpingof largerorganellewsithinthecytoplasm.Mitochondriacontainsome dilated cristaeT.en (10)peroxisomescounted. Z15905. Liver.Hepatocytewithclumpingof organellesT.he RER appearsdilated. Seven (7)peroxisomescounted. Z15906, Liver.Hepatocytewithhematopoieticellsalongtop,lefta,nd bottom.There isclumping of thelargerorganelles.Mitochondriacontainsome dense depositsand dilatedcristaeF.ifteen(15)peroxisomescounted.Some peroxisomesexhibitmarked degeneration. Z15907. Liver.Hepatocytemorphologyessentiallsyame asdescribedabove. Seven (7) peroxisomescounted. Z15908. Liver.Hepatocyteadjacentocentralvein.Appearssimilartopreviously describedcellswithclumpingofmitochondriand RER. Twelve (12)peroxisomes counted. Conclusiohs:Cellularfixatiosnuboptimaltopoor.There appearstobe slightly increasednumbers of peroxisomescompared withcontrolliver.Average 10.2 peroxisomes perhepatocyte. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.:418-01E3M-99.39 Animal No.: 19560-3 TreatmentGroup: - 1.6mg/kg Sex: -non ap,pli-cable Species/StrCrali:nCD:@BR VAF/PlusS Rat Tissue: Lung BlockNo(s).: 99.39-26 Z15910 - Photo/NegativNeo(s).: Z15914 SignificanLtesions(checkone): Yes InterpretinPgathologis(tsignaturaend date): x No Features:Z15910. Lung. Alveolusand liningcells.Allpneumocytesarecuboidalin thisphotograph.The secondcellfrom therightcontainsseverallamellarbodiesand isa PN2. The thirdcellalsocontainstwo smalllamellarbodies.No otherremarkable featuresnoted. Z1591 1. Lung. The darkcuboidallinincgellcontainsmultiplecytoplasmicvacuolesand appearstobe a PN2 althoughdefinitivleamellarstructureasrenotwithinthevacuoles. The pneumocyte attoprightalsocontains1-2vacuoleswhich probablyare/werelamellar bodies. Z15912. Lung. Alveolusand liningcells.Along thealveoluspneumocytesallappear flatteneadnd tobe PN Is. At bottom of photographincollapsedseptalareaisa cell containingmultiplelamellarstructurecsy,toplasmof a PN2. No lamellarstructureasre presentin the alveolarspace. Z15913. Lung. At topcentera largePN2 containsmultiplelamellarbodies.A darkthin PN I linestheremainderof theairwayspace.At bottom righta capillarcyontainsa large whitebloodcell.Dorsaltothisisan unidentifiecdellwithdilatedendoplasmicreticulum and nuclearenvelope. Z 15914. Lung. Alveolarspaceand liningcells.Withinthealveolusaremultiple extracellullaarmellarbodies.Liningpneumocytes inthisregionallappearto be PN Is. Severalcapillarieasrepresentinthephotograph. Conclusions: Essentiallnyormal fetallung.Some type11pneumocytes and lamellar bodiesidentifiedS.ome lamellarbodieswere occasionallpyresentinalveoli. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013EM-99.39 Animal No.: 19562-2 Treatment Group: 1.6mg/kg Sex: non apl)licable Species/StraCirln::CD@)BRVAF/Plus@ Rat Tissue: Liver Block No(s).: 99.39-27 Z15915 - Photo/NegativeNo(s).: Z15919 SignificantLesions (check one): X Yes No InterpretingPathologist(signatureand date): Features: Z15915. Liver. Hepatocyte and surrounding hematopoietic cells.Sinusoidal endothelium and plasma membranes are indistinctb,ut cytoplasmic organelles are fairly evenly dispersed. Dilated cristaeare present in the mitochondria. Nine (9)peroxisomes counted. Z15916. Liver.Hepatocyte and surrounding hematopoietic cells.Dilated cristaeare present in the mitochondria. Sinusoidal endothelium and plasma membranes are indistinct.Dilatationof RER. Eleven (11) peroxisomes counted. Z15917, Liver. Hepatocyte with clumping of organellesand washed-out appearance of much of the cytoplasm. Plasma membrane isindistinct.Twelve (12) peroxisomes counted. Z15918. Liver. Hepatocyte morphology similarto those described above. Mitochondria contain some dense deposits and dilatedcristae.Eight (8)peroxisomes counted. Z15919. Liver. Hepatocyte contains multiple clearvacuoles and marked dilationof RER. Mitochondria contain some dense depositsand dilatedcristae.Thirteen (13) peroxisomes counted. Conclusions: Poor cellularfixationof livertissue.There appears to be a slightincrease in the number of peroxisomes per hepatocyte. Average 10.6 peroxisomes per hepatocyte. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013EM-99.39 Animal No.: 19562-2 Treatment Group: -1.6 miz/kg Sex: - non ap]21icable Species/Strain:Crl:CDOBR VAF/Plus@ Rat Tissue-- Lung BlockNo(s).: 99.39-28 Z15920 - Photo/NegativeNo(s).:-Z 15924 SignificanLtesions(checkone): Yes InterpretinPgathologis(tsignaturaend date): X No Features: Z 15920.Lung. Alveolusand liningcells.At centerrightisa rounded liningcell which containstwo distincvtacuoles,and appearsto be a PN2. These appearconsistenwtith "empty" lamellarbody vacuoles.The liningcellson eithersideof thisround cellareflattened PNis. Similarlya,tleftcentrailsa cuboidalcellprotrudingintothelumen which containstwo vacuoleswhich appearedtobe empty lamellarbodies.The columnar cellatthe lower leftisa pneumocyte withoutremarkabledistinguishinfgeaturea,nd isprobablya PNI which hasn't flatteneodut. Z15921. Lung. Alveolusand collapsedsepta.At bottomrightisan ovalcellwhose cytoplasm containsone round vacuoleand multipleopen clefttshatappeartobe "empty" lamellarbodies. The darkoblongnucleusto itstopleftisan endotheliaclellofa capillarcyontaininga dark erythrocyteL.iningtheseptum on topof thisisa flattenePdNI. Anothercapillariysinthe centerof thecollapsedarea. Z]5922. Lung. Alveolusand liningcellsT.he centraldarkliningcellcontainsmany prominent mitochondriab,utalsocontainsa few vacuoleswhich may be lamellarbodies.A largerPN I is justtotheleftof thiscell.At thetopisan endotheliaclellnucleus. Z 15923. Luncr.In thecenterof thephotographisa scrolledlamellaorr myelin-likfeigurethat appearstobe extracellularwliythina capillarsypace. Dorsallyand ventrallayrecapillaries containinegrythrocyteasnd a lymphocyte(ventrallyT)o. itsrightisa liningPNI. Z15924. Alveolusand liningcells.A singledarktypeIIpneumocyte ispresentw,hich contains multiplefamellarbodies.Itisbounded on eithersideby flattenePdNIS. Capillarielsinedby endotheliaclellsand containinegrythrocyteasrepresentattoprightand lowerleft.No lamellar bodiesevidentwithinalveolarspaces. Conclusions: Essentiallynormal fetallung. Some random type IIpneumocytes containinglamellarbodieswere identifiedN.o lamellarbodieswere seen in alveolar spaces. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.:418-01E3M-99.39 Animal No.: 15962-3 TreatmentGroup: --1.6mg/kg Sex:-non aj2l2licable Species/StrCarli:nC:DOBR VAF/Plus@ Rat Tissue: Liver BlockNo(s).: 99.39-29 Z15925 - Photo/NegativNeo(s).: Z15929 SignificanLtesions(checkone): X Yes No InterpretinPgathologis(tsignaturaend date): Features: Z 15925 Liver.Hepatocytesurroundedby otherhepatocyteasnd a hematopoieticellon theright.There isclumpingof thecytoplasmicorganellesand largeareasofwashed outcytoplasm.Mitochondriaappeartohave some granular densitieasnd some dilatedcristae.There isdilatatioand blebbingof RER. Twenty-six (26)peroxisomescounted. Z15926. Liver.HepatocyteappearssimilartohepatocyteinZ15925. There are hematopoieticellson theleftand bottom. Twenty-three(23)peroxisomescounted. Z 15927, Liver.Hepatocytewithindistincyttoplasmicborders.There isdilatatioannd blebbingof RER. Mitochondriaappearto have some granulardensitieasnd some dilated cristaeT.wenty-one (21)peroxisomescounted. Z15928, Liver.Hepatocyteappearsessentiallsyimilartoabove cells.Fixationis suboptimalT.welve (12)peroxisomescounted. Z15929..Liver.Hepatocytewithindistinctytoplasmicborders.There isdilatationf RER. Mitochondriaappeartohave some granulardensitieasnd some dilatedcristae. Eighteen(18)peroxisomescounted. Conclusions: Increasednumbers ofperoxisomesperhepatocyte.Average 20.0 peroxisomesperhepatocyte.Poor,suboptimalfixation. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-013 EM-99.39 Animal No.: 19562-3 Treatment Group: -1.6 mg/kg Sex: - non ai)plicable Species/StraiCnr:l:CD@)BR VAF/Plus@ Rat Tissue: Lung BlockNo(s).: 99.39-30 Z15930 - Photo/NegativeNo(s).: Z15934 SignificantLesions (check one): Yes InterpretingPathologist(signatureand date): x No Features: Z15930. Lung. Alveolus and liningcells.Two PN2s with lamellarbodiesare present. One PN2 exhibitspseudopodia of cytoplasm extending intothe alveolus. The other PN2 contains severallargecytoplasmic vacuoles. Some small tubularsurfactantbodies are present in the alveolarlumen. Capillarieswith endothelialcellsand erythrocytesare present beneath the liningcells. Zi 593 1. Lung. Alveolus and liningcells.Centrallyisa dark PN2 which containsone distinct lamellarbody dorsalto itsnucleus. The cellimmediately on top of thisdark cellcontainsan open-faced nucleus but alsocontainsseverallamellarbodies,a PN2. The other liningcells appear to be PN Is. Z15932. Lung. Alveolus and liningcells.Within the alveolusisa freeand unattachedcell which isnot clearlyidentifiedas to celltype. All the liningpneumocytes appear to be flattened, but the one on the lower and middle rightappears to containsseveralvacuoles and one distinct lamellarbody, i.e.a, PN2 which has differentiateidntoa PNI. No lamellarbodies noted in alveolarlumen. Z15933. Lung. Alveolus and liningcells.The middle liningcellwith only a small portionof nucleus showing contains severalvacuoles,which appear to be "empty" lamellarbodies. The angular nucleus above itisa PN I nucleus. The largedorsalclearpneumocyte has not flattened and may contain a couple empty lamellarvacuoles. The oblong nucleus beneath the PN2 isan endothelialcellnucleus. Some small tubularstructuresin the lumen of the alveolusmay be releasedlamellarsurfactant. Z 15934. Lung. Alveolus,liningcellsand collapsedalveolarsepta. The liningpneumocytes all appear flattened.No lamellarbodies are present. At the top isa largeendothelialcellnucleus -and erythrocyteadjacentto it.No unusual findingsnoted. Conclusions: Several type 11pneumocytes with lamellar bodies seen. Also, some lamellar substance within alveolinoted. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013E;M-99.39 Animal No.: 19568-2 TreatmentGroup: -3.2miz/kg,high-dose Sex:-non arylicable Species/StrainC:rl:CDOBR VAF/Plus@ Rat Tissue: Liver BlockNo(s).: 99.39-11 Z15799 -Z15802, Photo/NegativeNo(s).Z:15909 SignificanLtesions(checkone): Yes -X No InterpretinPgathologis(tsignaturaend date): Features: Z15799 Liver.Hepatocytewithsinusoicdontaininhgematopoieticellat topand longitudinaslegment of bilecanaliculuastbottom right.Hepatocytecontainsone distincmtultivesiculabrody. Numerous mitochondriashow some slighdtilatioonf cristaea,nd thereissome dilatedblebbingof RER. Sinusoidalendotheliaclellattop is poorlydefined,suggestivoef suboptimalfixationF.ive(5)peroxisomescounted. Z15800. Liver.Hepatocytesurroundedby hematopoieticcells.Six (6)peroxisomes counted. Z15801. Liver.Hepatocytesurroundedby hematopoieticells.Plasma membrane of the hepatocyteisindistincatnd fixatioinspoor.. Mitochondriaappeardense withsome granulardepositsand dilatedcristaeD.ilatedblebsof RER arenoted.Three (3) peroxisomes counted. Z15802. Liver.SimilaroverallcellulamrorphologytoZ15801. Plasma membrane is indistinc.t.Mitochondriaappeardense withsome granulardepositsand dilatedcristae. Seven (7)peroxisomescounted. Z15909. Liver.Hepatocytewithgranularcytoplasmintermixedwithmitochondria, RER, and random peroxisomes.. Mitochondriaappeardense withsome granular depositsand dilatedcristaeN.ine (9)peroxisomescounted. Z15798: Liver.No negativen;o printdeveloped. Conclusions:Normal hepatocyte.SuboptimalfixationA.verage 6 peroxisomesper hepatocyte. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013E,M-99.39 Animal No.: 19568-2 Treatment Group: -3.2 mg/kg,-high-dose Sex:-non applicable Species/StraCriln::CD(bBRVAF/PlusS Rat Tissue: Lung BlockNo(s).: 99.39-12 Z15803 - Photo/NegativeNo(s).Z:15807 SignificanLtesions(checkone): Yes InterpretinPgathologis(tsignaturaend date): x No Features: Z15803. Lung. Air spacelinedby severalcuboidalepitheliaclells.These cellsappear tobe pneumocytes which have not flattenedout intodifferentiatePdN Is. They have finegranularcytoplasmwithoutmany distincotrganelles.Dorsallyaretwo blood vesselslinedby endotheliaclells.No lamellarbodiesnoted. Z 15804. Lung. Alveolusand liningcells.Two cuboidalpneumocytes arePN2s and containlamellarbodies.Two adjacentPNIs areassociatedwith thesecells.Adjacent tissueiscollapsedseptaand celltypesarelessclearlydefinable. Z15805. Lung. A clusterof cuboidalpneumocytes sin-tiltaorthosedescribedinZ15803. To the leftisa capillarylinedby an endotheliaclell.Within thecapillarayre two lamellar-liksetructuretshatappeartobe extracellularN.o PN2s or alveolarlamellar bodies noted. Z15806. Lung. Alveolus withliningcells.Pneumocytes arelow cuboidaland appear to be PN Is,althoughnotcompletelyflatteneodut. No PN2s noted. The celltothe right with an oblong nucleus appearstobe a fibroblastI.tcontainsnumerous RER and some coliagenfibrilasrepresentextracel,'ulary. Z15807. Lung. Alveoluswith two PNIS, althoughcellsarenotflattenedA.djacent tissueiscollapsedseptaand interstitiuNmo. PN2s or alveolarlamellarbodiesnoted. Conclusions: Occasionaltype 11pneumocytes noted. Some intravasculalramellar-like materialalsopresent.Many typeIpneumocytes have apparentlynot flatteneodut, probably normal fetallung. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013E,M-99-39 Animal No.: 19569-1 Treatment Group: 3.2mg&g, high-dose Sex:-non al2plicable Species/StrainC:rl:CDOBR VAF/PlusS Rat Tissue: Liver BlockNo(s).: 99.39-13 Z15843 - Photo/NegativeNo(s).-:Z 15847 SignificanLtesions(checkone): X Yes No InterpretinPgathologis(tsignaturaend date): Features: Z15843. Liver. Hepatocyteadjacentocentralvein.Abundant numbers of mitochondria and RER arepresent,but plasma membranes and sinusoidalendothelium arepoorlydelineated.Cristaearedilatedand broken up in some mitochondria. Multivesiculabrody present. Eleven (11)peroxisomescounted. Z15844. Liver.Hepatocytewith some rarefactioonfcytoplasm. DilatedRER are evidentand some granulardepositsarepresentinmitochondria.Eleven (11) peroxisomes arecounted. Z15845. Liver.Hepatocytewith hematopoieticcellinsinusoidatlower leftof photograph. Mitochondriain hepatocytecontaindilatedcristaeT.wo multivesicular bodiesarepresentinthecytoplasm. The RER appearsdilatedwith some blebbing. Cytosol appearsrarefiedand plasma membrane ispoorlydef-inedE.leven (11) peroxisomes counted. Z15846. Liver.Hepatocytesurroundedby hematopoieticells.Morphology similarto thosedescribedabove. A golgiassociatedwith multivesiculabrody ispresent.The mitochondriacontainmultiplegranulardepositsand dilatedcristae.Eleven (11) peroxisomes counted. Z15847. Liver. Hepatocyte,similartoabove descriptionsP.lasma membrane ispoorly definedand fixationissuboptimal.The mitochondriacontainmultiplegranulardeposits and dilatedcristae.Nine (9)peroxisomescounted. Conclusions: Hepatocytesappeartocontainslightlmyore peroxisomes. Average 10.6 peroxisomes per hepatocyte.Suboptimalfixation.No otherdifferencenotedcompared with control. I TRANSMISSION ELECTRON MICROGRAPH Study No.: -418-013: EM-99.39 Animal No.: 19569-1 Treatment Group: 3.2 mg/kg, high-dose Sex:-non al2l)licable INTERPRETATION Species/StrainC:rl:CDOBR VAF/PlusS Rat Tissue-: Lung BlockNo(s).: 99.39-14 Z15848 - Photo/NegativeNo(s).:Z15852 SignificanLtesions(checkone): Yes -X No InterpretinPgathologis(tsignaturaend date): Features: Z15848. Lung. Alveoluswith liningcells.Severalcuboidalpneumocytes are present.The centralcellhas two basilarvacuoleswhich probablyrepresentlamellar bodies,althoughthelamellarmaterialisabsentinthem. A smallportionof a septal capillaryispresentat left. Z15849. Lung. Sectionshows severalcuboidaltocolumnar epitheliaclells.The central cellappearstocontaintwo smallvacuoles,one with some lipid-likmeaterialp,ossiblya lamellarbody. To thelower rightof thiscellisa cellwithan irregulanrucleusand severallipidor lamellarvacuoles;thiscellappearstobe a macrophage. A capillaryisin the centerof thephotograph with an endothelialiningcells. Z 15850, Lung. A clusterofcuboidalpneumocytes,with mostly cytoplasmcontaininga finegranularmaterial.No characteristliacmellarbodiesarepresentinthesepneumocytes or inthealveolarspace.An interstiticaellcontainsa singlemyelin-likeor lamellarbody, but thiscellisnot clearlyidentifieads tocelltype. Z15851.Lung. Two PN2s arepresentand theycontainmultiplelamellarbodies.A fibroblasatppearstocontaintheelongatednucleustotherightof thesePN2s. Z15852. Lung. A clusterof pneumocytes orepitheliaclellsas describedinabove photographs.The topcellcontainsa couplelamellarbodiesand isa PN2. The cells appeartobe pneumocytes (PN Is)which haven'tflatteneodut,or PN2s intransitiotno PNIS. Conclusions: Essentiallynormal lung. Some random PN2s were identifiedT.here are numerous cuboidalpneumocytes which appear to not have flattenedout. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.:418-013E,M-99.39 Animal No.: 19572-1 TreatmentGroup: -3.2mg/kg, high-dose Sex:-non apt)licable Species/StrainC:rl:CDOBR VAF/Plus@ Rat Tissue: Liver BlockNo(s).: 99.39-15 Z15853 - Photo/NegativNeo(s).:-Z15857 SignificanLtesions(checkone): X Yes No InterpretiPnagthologis(tsignaturaend date): Features: Z15853. Liver.Plasma membranes arepoorlydefinedand many cytoplasmic detailasreapparentlylost.Membranes and cristaweithinmitochondriaareindistinct. Nine (9)peroxisomescounted. Z15854. Liver.SimilartoZ15853,plasma and cellulamrembranes arepoorlydefined due topoor fixationR.ER appearsmarkedly dilatedT.hirteen(13)peroxisomescounted. Z15855. Liver.Hepatocytewithhematopoieticellstotherightmarginand another hepatocyteand bilecanaliculuastthetop.RER profileasredilatedand plasma membranes areindistincTtw.elve (12)peroxisomescounted. Z15856. Liver.Hepatocellulamrorphology issimilartocellsdescribedabove. Fourteen (14)peroxisomescounted. Z 15857. Liver. Hepatocyteand surroundinghematopoieticells.Plasma and cellular membranes arepoorlydefined.Eighteen(18)peroxisomescounted. Conclusions: Hepatocytespoorlyfixed.There arehighernumbers ofperoxisomes withinhepatocytesascompared withcontrols.Average 13.2peroxisomesper hepatocyte. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-013;EM-99.39 Animal No.: 19572-1 TreatmentGroup: -3.2mg/kg, high-dose Sex:-non applicable Species/StraiCnr:l:CDOBR VAF/Plus@ Rat Tissue-:Lung BlockNo(s).: 99.39-16 Z15858 - Photo/NegativNeo(s).:-Z15863 SignificanLtesions(checkone): Yes InterpretinPgathologis(tsignaturaend date): X No Features: Z15858. Lung. AlveolusandlinincgellsC.entrallinincgellappearstocontainone lamellarbody totherightofitsnucleusa, PN2. To itsleftisa capillarcyontaininagn erythrocytaend liningendotheliaclell.Plasmaand cellulamrembranes arepoorlydefineddue to suboptimalfixationS.ome celluladrebrisispresentinthealveolasrpace. Z 15859.Lung. AlveolusandpredominantlcyuboidalinincgellsL.iningcellon rightand bottomrightbothcontainsome lamellabrody vacuolesand appeartobe PN2s. The othertwo pneumocytesappeartobe PNIS. No specifiacbnormalitiensoted. Z15860. Lung. Collapsedseptumand liningcells.Most appeartobetypeIpneumocytes, althoughnotflatteneodut.No lamellarbodiesidentifieFdi.xationispoor. Z15861. Lung. Largetopcentercellisa pneumocyte.At itsdorsalmarginisvacuolethatmay be an "empty"lamellabrody.Otherlinincgells.appetaorbe typeIpneumocytes.At bottom rightisa smallportionofa cellwithsome lysosomesthatappeartocontainsome crystalosr possiblylamelladrebris.Thiscellmay be a PN2 ora macrophage.The cellattoprightwith numerous cytoplasmicinclusionissa granulocytewithina capillaryi,tsendothelianlucleus directlbyelow thegranulocyte. Z15862. Lung. Collapsedsepta.Liningthisareseverafllattenepdneumocytes,bothdarkand light.The collapsedareacontainseveralight-stainpinegumocytesand endotheliaclells.A granulocyteispresentinthecapillarayttheright.The ovalnucleusnexttothisvesselappearsto belongtoa fibroblastN.o abnormalitiensoted.No lamellabrodiesclearleyvidentinthis photograph. Z15863.Lung. Collapsedseptalarea.The lamellamryelin-likmeaterialinthetopcenteris withina capillarlyumen. ne topairspaceislinedby a flattenePdNI. Some lamellamraterial may be presentinthealveolus.At thebottomleftisa darkercellwhich containsa lamellar body. Conclus ons: Essentiallynormal fetallung.Some lamellarbodies noted in type Il pneumocytes. Fixationwas suboptimal. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013E:M-99.39 Animal No.: 19577-4 Treatment Group: 3.2 mg@kg, high-dose Sex: -non aj2]21icable Species/StraCirnl:CDOBRVAF/PlusS Rat Tissue: Liver Block No(s).:-99.39-17 Z15864- Z15868 Photo/NegativeNo(s).: Z15940 SignificantLesions (check one): X Yes No InterpretingPathologist(signatureand date): Features: Z15864. Liver. Hepatocyte with surrounding erythrocytesin sinusoids. Plasma and cellularmembranes are indistincdtue to poor fixation.Mitochondria show some granular depositsand dilatedcristae.Three (3)peroxisomes counted. Z 15865. Liver. Hepatocyte surrounded by adjacent hepatocytes and hematopoietic cells. A bilecanaliculusispresent at the bottom of the photograph. Eleven (11) peroxisomes counted. Z15866. No negative. Z15867. Liver. The plasma membrane isindistinct.Eleven (11) peroxisomes counted. Z15568. Liver. Hepatocyte and adjacent hematopoietic cells.Dilationand blebbing of RER. Plasma membranes are indistinct.Sixteen (16) peroxisomes counted. Some of the peroxisomes show advanced degeneration. Z15940. Liver. Hepatocyte with poorly-defined plasma membranes. Mitochondria show dilatationof cristae.Fifteen(15) peroxisomes counted. Conclusions: Hepatocytes exhibitfeaturesof poor fixation.There appears to be a slight increase in the number of peroxisomes per hepatocyte. Average 11.2 peroxisomes per hepatocyte. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.: 418-013E,M-99-39 Animal No.: 19577-4 TreatmentGroup: 3.2mg/kg, hi-gh-dose Sex:-non a]pl2licable Species/StrCarli:nC:D@)BRVAF/PlusS Rat Tissue-:Lung BlockNo(s).: 99.39-18 Z15869 - Photo/NegativNeo(s).:Z15873 SignificanLtesions(checkone): Yes X No InterpretinPgathologis(tsignaturaend date): Features: Z15869. Lung. Alveolarspaceand liningcells.Centralpneumocyte on rightcontainsseverallamellarbodiesand isa PN2. A detachedcellisjustabove thisone and may be a pneumocyte or possiblya lymphocyte.At thetopleftisa capillarwyith an oblong endotheliaclellnucleus. Z15870. Lung. Area of collapsedalveolarsepta.Centrallyisa darkcellthatcontains severalcytoplasmicvacuoles,two of which containlamellarbody material.Two cell beneaththiscellalsocontainsimilarcytoplasmicvacuolesand may be PN2s. Other cells with abundantmore clearcytoplasmappeartobe typeI pneumocytes,althoughtheyare not flattened. Z 15871. Lung. Ventrallya PN I linestheairwayspace.The smallerdarkcelladjacentto and coveredby itcontainsmultiplesmallclearvacuoles,butlamellarmaterialisnot clearlyevident.Dorsallythetissuecontainscollapsedseptawitha mixtureof endothelial cellsc,apillarlyumen, pneumocytes,and interstitiuNmo. abnormalitiensoted. Z15872. Lung. Area ofcollapsedalveoliand septa.Centrallyaretwo PN2s containing vacuolesand lamellarbodies.Ventrallyisa capillarcyontaininga lamellar-typbeody withinitslumen. Z15873. Lung. Ventrallyareseveralcuboidalpneumocytes liningan alveolarspace. The centralcellcontainsa couplesmalllamellar-typsetructureisnitscytoplasm.To the rightand top right are capillaries containing some lamellar material in their lumina. At the top is a flattenedpneumocyte. Conclusions: Normal lung. Some type 11pneumocytes identified.Some lamellar structurespresent within capillarylumina. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.:418-01E3M-99.39 Animal No.: 19579-3 Treatment Group: 3.2mg/kg, high-dose Sex:-non ai)121icable Species/StrainC:rl:CDOBR VAF/Plus@ Rat Tissue: Liver Block No(s).: 99.39-19 Z15874 - Photo/NegativeNo(s).: Z15878 SignificantLesions (check one): X Yes No InterpretinPgathologist(signatureand date): Features: Z15874. Liver. Hepatocyte surrounded by hematopoieticcellat lower left and sinusoidcontainingerythrocytesat upper right.Endothelialcellispoorly delineated and hepatocellularplasma membranes areindistinctS.eventeen (17) peroxisomes counted. Z15875. Liver. Hepatocyte mixed with some hematopoieticcells.Hepatocellular boundaries are incomplete on thiscell,and peroxisomes are not counted for thiscell. Granular depositsare presentin some mitochondria. There appears to be abundant rough endoplasmic reticulum. Z15876. Liver. Hepatocyte with indistincptlasma membranes, but containsabundant mitochondria,endoplasmic reticulum,and peroxisomes. Thirty(30)peroxisomes counted. Z15877. Liver. Hepatocyte,similarin overallmorphology to those describedabove. Some granulardepositsarepresentwithinsome mitochondria. Endoplasmic reticulum and nuclearenvelope appear dilated.Twenty-four (24)peroxisomes counted. Z15878. Liver. Hepatocyte,similarin overallmorphology to those describedabove. Some granulardepositsarepresentwithinsome mitochondria. Endoplasmic reticulum and nuclearenvelope appear dilated.Sixteen(16)peroxisomes counted. Conclusions: Tissue fixationsuboptimal,but betterthan some previousanimals.There appears to be increasednumbers of peroxisomes per hepatocyte. Average 21.75 per hepatocyte(fourcellscounted). No otherchanges noted compared with controllivers. TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION StudyNo.:-4-18-013EM-99.39 Animal No.: 19579-3 TreatmentGroup: 3.2mg/kg, hi-gh-dose Sex:-non applicable Species/StraCirnl::CDOBR VAF/PliisS Rat Tissue: Lung BlockNo(s).: 99.39-20 Z15879 - Photo/NegativNeo(s).:-Z-15883 SignificanLtesions(checkone): Yes InterpretinPgathologis(tsignaturaend date): x No Features:Z15879. Lung. Alveolus,linincgellsandcollapseadlveolasreptaarea.The lining cellcontainsseveralamellabrodiesand isa PN2. Some miscellaneoucselluladrebrisappears tobe inthealveolarlumen,butno lamellarmaterialT.o thefightisa cellcontaininsge,veral cytoplasmicvacuolesand an irregulanrucleus.Thismay be a PN2, butalsomay be a macrophage. Othercellsappeartobe compressedpneumocytes. Z15880.Lung. Centralliysa finegranularmateriaclontaininmgultiplelamellarbodies.This appearstobe extracellulalrt,houghitappearsmixed withothercelluladrebrisand isprobablya ruptured(artifactuallPyN?2). Othercellslinintgheairwayappeartobe PN Is. Z]5881. Lung. Area ofcollapseadlveolaind septa.Centrallayreseveralcellswhich contain largevacuolescontainingsome scantmaterialT.hese may be PN2s, butalsomay be macrophages.At thetopan ellipticvaalcuoleappearstocontainsome lamellarmaterialT.hree dorsalnucleihave dilatednuclearenvelopess,uggestinsguboptimalfixation. Z15882.Lung. Thickened,collapsesdeptalwall,linedby pneumocytes.Some freelamellar materiaalppearsinthealveolalrumen on theleftmixed withsome celluladrebris.The central cellinthe"septum"appearstobe a PNI. The fightmarginislinedby a PN I.The darkseptal cellatthelowerrightcontainsome lamellabrodiesand isprobablya PN2. Z]5883. Lung. Area ofcollapseadlveolaind septa.SimilartoZ15880,thismightrepresenat rupturedPN2. At topinthelumen ismultiplefamellabrodies.The collapsedseptaand capillariaepspearessentialnloyrmal. Conclusions: Essentiallynormal lung. Some normal type 11pneumocytes identified. Some lamellarmaterialidentifiewdithinalveoli. Ell.TransmissionElectronMicrographs NOTE: The originagllossyfigurephotographs(FiguresI - 14)were deliveredto Dr. Marvin T. Case with thedraftpathologyreporton March 19, 1999. FigureLegends AncillarPyathologRyeport PfimedicAargusNo. 418-013 MagnificationP.rinmtagnificatiiosinndicatoend thebackofeachphotographA.ll reportfiguresare ata magnificationof 11,20OX. The measurement bar on each photomicrographequalsone micron (0.001mm). FigureI. Liver.Hepatocyteshowing normal cytoplasmicorganelles.Clumping of mitochondriaand rough endoplasmic reticulumreflectssuboptimalfixation.Some peroxisomes areidentifiedH.ematopoieticcellsarepresentventrallyA. bilecanaliculus ispresentatthejunctionof threehepatocytesinthetop left.Animal No. 19506-4,0 mg/kg/day (control)1,1,20OX magnification. Figure2. Liver.Hepatocyte.Dilatatioonf therough endoplasmicreticuluma,n indistincptlasma membrane, and organelleclumping indicatesuboptimalfixation. Mitochondria aredense and show some granulardepositsand dilatedcristae.Some representativpeeroxisomes arelabeled.An endothelialcellnucleus ispresentliningthe sinusoidin theupper leftcomer of thephotograph. Animal No. 19509-1,0 mg/kg/day (control)I,1,20OX magnification. Figure3. Liver.Cellularmorphology issimilartoFigure2. Some peroxisomesare labeledand appearmore numerous thanincontrols.The rough endoplasmic reticulum and nuclearenvelope aredilated.A bilecanaliculusispresentin upper leftof the photograph Animal No. 19534-1,0.4 mg/kg/day,11,20OX magnification. Figure4. Liver.Mitochondriashow dilatatioonf cristae.Some peroxisomes,increased innumber, arelabeled.Some hematopoieticcellsarepresentinthe adjacentsinusoid. Animal No. 19560-2,1.6mg/kg/day, 11,20OX magnification. Figure5. Liver.Hepatocytewith an indistincptlasma membrane. Cristaeare dilated withinmitochondria.Severalperoxisomesare labeled.Animal No. 19577-4,3.2 mg/kg/day (high-dose)1,1,20OX mao0nification. Fi.gure61 Liver. Hepatocyte whose plasma membrane isindistinctR.epresentative peroxisomes arelabeled.Animal No. 19579-3,3.2mg/kg/day (high-dose),11,20OX magnification. Figure7. Lung. Alveolusand liningcells.Two type Ilpneumocytes containinglamellar bodiesare present.The centraldarkcellmay be a typeIIpneumocyte which has expresseditslamellarbodies.Type I pneumocytes can be seen liningthe alveolusatthe top and bottom margin of theairspace. Subjacentalveoliarecollapsedand the representedcellsarepneumocytes and septa]cellsatdifferenftunctionsofcut. Animal No. 19505-1,0 mg/kg/day (control)1,1,20OX magnification. AncillaryPathologyReport PrimedicaArgus No. 418-013 Fijzure8. Lung. Alveoliand septum. A typeH pneumocyte ispresentliningthe right alveolus.This cellcontainsseverallamellarbodiescontainingfaintlaminatedmaterial. The otherliningcellsaretypeI pneumocytes. An endothelialcelland capillarayre presentinthe septum atthe topof thephotograph. Animal No. 18915-1,0 mg/kg/day (control)1,1,20OX magnification. Figure9. Lung. Alveolus and septa.The centralnarrow alveolusisbounded by two septacontainingcapillariewsith luminalerythrocytesA. lamellarmyelin-likestructuries presentin thelumen of the upper capillary.FlattenedtypeI pneumocytes linethe alveolus.No typeIlpneumocytes or lamellarbodiesareevidentinthisphotograph. Animal No. 19504-3,0 mg/kg/day (control)1,1,20OX magnification. Figure10. Lung. Alveolusand liningcells.A typeIIpneumocyte containingmultiple lamellarbodies ispresentcentrally.Much ofthe lamellarmaterialhas washed-out of the lamellarbodies. A second type IIpneumocyte ispresentnearthebottom, and containsa singlelamellarbody. The remainingcellsappeartobe typeI pneumocytes. Animal No. 19546-1,0.4 mg/kg/day, I1,20OX magnification. Figure 11. Lung. Alveolusand liningcells.At thetopisa cuboidaltype IIpneumocyte containingseverallamellarbodies. The adjacentliningcellsappeartobe pneumocytes which have not flatteneodut. The lamellarmyelin-likestructureasre extracellulawrithin capillaryspaces. Animal No. 19560-2,1.6mg/kg/day, 11,20OX magnification. Figure 12. Lung. Alveolusand liningcells.Alveolarspacecontainsabundant lamellarlikematerial,which isconsistenwtith surfactanstecretedfrom typeIlpneumocytes. The dark cellularsegment nearthebottom may be an extensionof a type11pneumocyte. The remaining liningcellsappeartobe typeI pneumocytes orcuboidalpneumocytes. Capillariewsith liningendotheliaclellsarepresentdorsallyand atthelower right. Animal No. 19560-3,1.6mg/kg/day (high-dose)I,1,20OX magnification. Figure13. Lung. Alveolusand liningcells.Two type IIpneumocytes containing lamellarbodies arepresent.A typeI pneumocyte ispresentliningthealveolusventralto thetype IIpneumocytes. Within theseptalwallcollagenfibersand a fibroblasatre evident.The materialinthealveolusisnondescriptcelluladrebris.Animal No. 19569-4, 3.2 mg/kg/day (high-dose),11,20OX magnification. Figure 14. Lung. Alveoliand septum. Both sidesof theseptum are linedby flattened pneumocytes. The largecentralcellinthe septum appearstobe a type Ipneumocyte. Two cellsventrallycontainlamellarbodiesand aretype11pneumocytes. Animal No. 19579-3,3.2 mg/kg/day (high-dose),11,20OX magnification. Abbreviation A BC BE CL Cap CN co CP D DD DC DN DR EC ER FB G HC L LB m N p PNI PN2 PM RER AncillaryPathology Report PrimedicaArgus No. 418-013 Index of Labels for Ultrastructural Structures Structure Alveolar space Bile canaliculus Bronchiolar epithelium Capillarylumen Capillary Centrioles Collagen Cuboidal pneumocyte Desmosome Dense deposits Dilated cristae Dilated nuclear envelope Dilated RER Endothelialcell Erythrocyte Fibroblast Glycogen Hematopoietic cells Lipid droplet Lamellar body Mitochondria Nucleus Peroxisome Type I pneumocyte Type IIpneumocyte Plasma membrane Rough endoplasmic reticulum ri O'D7c DD.: - cc 14 !kA"@ lo' DC p R Bc A, I-* ism P7 01 41. kr io 41 "6.4 4t Ir ;Pt, op 4L 14 4ri t@R--- --ft 7g. Ilk 46, PM ,03 10 .ere ep iRr. 10 IL A..Ll e3 60 Ir it 'Sid I I ZL; ip 44 LA VV I'll 360 Ilk LB PN A PN1 .4t IL tit 4k4 1 PN1 LB A Is F LB A PN1 or fir sr MOE 41 LB B CA 41 :IV. A 4@ J'At. Ilk' It PN 4r' -CP LB T-W L PN1 Ole lk lb @AIOL 41 PN c N 2,. CO PN1 LB A PLI J;.v p 1. pk le PNl@ A 'Ilk IL p L P- IV. QualityAssuranceStatement Patholgoy Associateslntemational A CompanoyfScienAcpeplicatIinotnesrnatCioornpaolration -&-@ mosompOF-Evarumr%mr& An Emplovee-O@ne0d0-canv QUALITYASSURANCESTATEMENT This electronmicroscopystudyhas been inspectedand auditedby the Quality Assurance Unitas requiredby the Good LaboratoryPracticesRegulationspromulgated by the U.S. Food and Drug Administration.PAI has a functioninagnd responsive QualityAssurance Unitwhich reportsdirectltyo management. The followingisa record ofinspections/auditasnd theirresultinrgeports: Date of Inspection Phase Inspected Date FindingsRel2orted to Managemen and Study Directo 2/22/99 Processingand Embedding 3/22,23, 24, 25/99 Data Audit 3/25/99 DraftReportAudit 4/5/99 FinalReport Audit 2/22/99 3/25/99 3/25/99 4/5/99 2@aeann@nee@dWeawradr,d, B.S.,ROAP-GLP Date QualityAssurance Specialist Sponsor: 3M Corporate Toxicology Test Article:PFOS Study Number: 418-013, PAI Study EM-99.39 4915 D ProspectusDrivea Durham, North Carolina27713 (919)544-5257 (919)544-3218 FAX