Document 44xQb5qmm4zGxMp7Xz6mBvM1N

FINAL REPORT /H&36-0553 received OPPT CBIC 2000FEB 22 AM||; 3* PROTOCOL 418-014 ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS SPONSOR'S STUDY NUMBER: T-6295.13 FINAL REPORT DATE: 23 JULY 1999 # 002S9- PROTOCOL 418-014 ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS SPONSOR'S STUDY NUMBER: T-6295.13 TABLE OF CONTENTS SUBJECT I. SUMMARY AND CONCLUSION A. Methods B. Results C. Conclusion II. DESCRIPTION OF TEST PROCEDURES A. Conduct of Study A.1. Sponsor A.2. Testing Facility A.3. Study Number A.4. Sponsor's Study Number A.5. Purpose of the Study A.6. Study Design A.7. Regulatory Compliance PAGE 1-1 1-1 I-4 I-6 11-1 11-1 11-1 11-1 11-1 11-1 11-1 11-1 11-1 omrzso J l# 3 SUBJECT A.8. Ownership of the Study A.9. Study Monitor A.10. Alternate Study Monitor A.11. Study Director A. 12. Technical Performance A.13. Report Preparation A.14. Report Review A.15. Date Protocol Signed A.16. Dates of Technical Performance A. 17. Records Maintained B. Test Article Information B.1. Description B.2. Lot/Batch Number B.3. Date Received and Storage Conditions B.4. Special Handling Instructions B. 5. Analysis of Activity C. Vehicle Information C.1. Description C.2. Lot Numbers C.3. Dates Received and Storage Conditions C.4. Special Handling Instructions C.5. Analysis of Purity PAGE 11-2 11-2 II-2 II-2 II-2 II-2 II-2 II-2 II-3 II-3 II-3 II-3 II-3 II-3 II-4 II-4 II-4 II-4 II-4 II-4 II-4 II-4 iMT W 0U .i9.JL SUBJECT D. Test Article Preparation and Storage Conditions D.1. Sample Information D.2. Analytical Results E. Test System E.1. Species E.2. Strain E.3. Supplier (Source) E.4. Sex E.5. Rationale for Test 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 Facility Registration F.2. Study Rooms F.3. Housing F.4. Lighting F.5. Sanitization F.6. Feed F.7. Feed Analysis F.8. Water m PAGE II-5 II-5 II-5 II-5 II-5 II-5 II-6 II-6 II-6 II-6 II-6 II-6 II-7 II-8 II-8 II-8 II-8 II-8 II-8 II-8 II-9 II-9 Q00S59 g SUBJECT F.9. Water Analysis F.10. Bedding F. 11.Bedding Analysis G. Methods G.1. Dosage Administration G.2. Rationale for Dosage Selection G.3. Route of Administration G.4. Rationale for Route of Administration G.5. Frequency of Administration G.6. Length of Study G. 7. Method of Study Performance H. Gross Necropsy H.1. Fo Generation Rats H. 2. F1 Generation Pups I. Statistical Analyses III. RESULTS A. Mortality, Clinical and NecropsyObservations B. Body Weights C. Absolute (g/day) and Relative (g/kg/day) Feed Consumption Values D. Natural Delivery and Litter Observations D.1. Natural Delivery Observations D.2. Cross-Fostering Litter Observations IV PAGE 11-9 II-9 II-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 III-2 III-2 IH-2 III-2 * * * * * Mv\H SUBJECT E. Clinical Observations from Birth to Day 21 Postpartum and Necropsy Observations PAGE III-3 F. Electron Microscopic Evaluation of the Liver and Lung from Day 1 Postpartum Pups III-4 F.1. Liver III-4 F.2. Lung III-4 REFERENCES III-5 APPENDIX A - REPORT FIGURES Figure 1. Maternal Body Weights A-1 Figure 2. Maternal Body Weights - Rats Assigned to Cross Fostering A-2 APPENDIX B - REPORT TABLES Table 1. Clinical Observations - Summary - Fo Generation Female Rats B-1 Table 2. Necropsy Observations - Summary - Fo Generation Female Rats B-4 Table 3. Body Weights - Precohabitation - Summary - Fo Generation Female Rats B-5 Table 4. Body Weight Changes - Precohabitation - Summary Fo Generation Female Rats B-6 Table 5. Maternal Body Weights - Gestation - Summary Fo Generation Female Rats B-7 Table 6. Maternal Body Weight Changes - Gestation - Summary Fo Generation Female Rats B-8 Table 7. Maternal Body Weights - Lactation - Summary Fo Generation Female Rats B-9 Table 8. Maternal Body Weight Changes - Lactation - Summary Fo Generation Female Rats B-10 v 000254 250 SUBJECT Table 9. Absolute Feed Consumption Values (g/day) Precohabitation - Summary - Fo Generation Female Rats PAGE B-11 Table 10. Relative Feed Consumption Values (g/kg/day) Precohabitation - Summary - Fo Generation Female Rats B-12 Table 11. Maternal Absolute Feed Consumption Values (g/day) Gestation - Summary - Fo Generation Female Rats B-13 Table 12. Maternal Relative Feed Consumption Values (g/kg/day) Gestation - Summary - Fo Generation Female Rats B-14 Table 13. Maternal Absolute Feed Consumption Values (g/day) Lactation - Summary - Fo Generation Female Rats B-15 Table 14. Maternal Relative Feed Consumption Values (g/kg/day) Lactation - Summary - Fo Generation Female Rats B-16 Table 15. Natural Delivery Observations - Summary - Fo Generation Female Rats B-17 Table 16. Litter Observations (Naturally Delivered Pups) - Summary - F1 Generation Litters B-20 Table 17. Clinical Observations from Birth to Day 21 Postpartum Summary - F1 Generation Pups B-24 Table 18. Necropsy Observations - Summary - F1 Generation Pups B-25 Table 19. Clinical Observations - Individual Data - Fo Generation Female Rats B-26 Table 20. Necropsy Observations - Individual Data - Fo Generation Female Rats B-30 Table 21. Body Weights - Precohabitation - Individual Data Fo Generation Female Rats B-33 Table 22. Maternal Body Weights - Presumed Gestation - Individual Data - Fo Generation Female Rats B^5 Table 23. Maternal Body Weights - Lactation - Individual Data Fo Generation Female Rats B-53 16V U ' . /U\ Ui J r l * i 4MB SUBJECT Table 24. Feed Consumption Values - Precohabitation - Individual Data - Fo Generation Female Rats PAGE B-59 Table 25. Maternal Feed Consumption Values - Presumed Gestation - Individual Data - Fo Generation Female Rats B-63 Table 26. Maternal Feed Consumption Values - Lactation - Individual Data - Fo Generation Female Rats B-71 Table 27. Natural Delivery, Implantation Sites, and Pup Viability and Sex - Individual Data - Fo Generation Female Rats/ F1 Generation Litters B-74 Table 28. Duration of Gestation and Average Duration of Delivery (Minutes) Per Pup Per Litter - Individual Data Fo Generation Female Rats B-77 Table 29. Pup Body Weight Litter Averages from Birth to Day 21 Postpartum - Individual Data - F1 Generation Litters B-80 Table 30. Pup Body Weights from Birth to Day 21 Postpartum Individual Data - F1 Generation Pups B-83 Table 31. Pup Vital Status and Sex from Birth to Day 21 Postpartum Individual Data - F1 Generation Pups B-99 Table 32. Clinical Observations from Birth to Day 21 Postpartum Individual Data - F1 Generation Pups B-102 Table 33. Necropsy Observations - Individual Data - F1 Generation Pups B-104 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 AND RELATIVE HUMIDITY REPORTS E-1 to E-7 APPENDIX F - PATHOLOGY REPORT F-1 to F-46 APPENDIX G - STATEMENT OF THE STUDY DIRECTOR G-1 VII 252- SUBJECT APPENDIX H - QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT PAGE H-1 to H-5 l*>3 Vili w in 418-014:PAGE 1-1 TITLE: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS ARGUS RESEARCH LABORATORIES, INC. PROTOCOL NUMBER: 418-014 SPONSOR'S STUDY NUMBER: T-6295.13 I. SUMMARY AND CONCLUSION A. Methods3 Seventy-eight virgin female Crl:CDBR VAF/Plus (Sprague-Dawley) rats were assigned to two dosage groups (Groups I and II), 42 rats in the control group and 36 rats in the treated group. The rats were administered the test article, PFOS (FC-95), or the vehicle, 0.5% Tween 80, orally (via gavage), once daily beginning 42 days before cohabitation through day 21 postpartum (DL 21). The dosage volume was 5 mL/kg. Dosages were adjusted daily for body weight changes and given at approximately the same time each day. Rats were assigned to cross-fostering or pharmacokinetic sample collection depending on the order of delivery and the availability of rats for cross-fostering. Twenty-five litters per dosage group were assigned to cross-fostering on DL 1. The litters were assigned such that four subsets were obtained among the two dosage groups as described in the table below. Dosage Group I I II II Number of Mated Fem ale Rats 13 12 12 13 Dosage (m g/kg/day) 0 (Vehicle) 0 (Vehicle) 1.6 1.6 Number of Litters R e a s s ig n e d 13 12 12 13 Reassigned Litter Type (Dam Treatment) Treated Untreated Treated Untreated Subset Number A B C D Eight female rats from the control group and two female rats from the treated group were assigned to pharmacokinetic sample collection following delivery. These rats were not cross-fostered. a. Detailed descriptions of all procedures used in the conduct of this study are provided in the appropriate sections of this report and in APPENDIX C (PROTOCOL AND AMENDMENT). O O O S S L fflW 418-014:PAGE I-2 A.1. Fo Generation Rats Assigned to Cross-Fostering: The Fo generation female rats were observed for viability at least twice each day of the study and for clinical observations of effects of the test article, abortions, premature deliveries and/or deaths before and approximately one hour postdosage. Body weights were recorded daily during the dosage period and at sacrifice. Feed consumption values were recorded weekly to cohabitation, daily during the gestation period and on DLs 1, 4, 7, 10 and 14. The female rats were evaluated for duration of gestation, length of parturition, litter sizes and pup viability at birth. Pups that either appeared stillborn or that died before initial examination of the litter for viability were examined for vital status at birth. Maternal behavior of the dams was evaluated daily when the pups were examined during the 21-day postpartum period. Observed maternal behavior was recorded on DLs 1,4, 7, 14 and 21. Twenty-four-hour urine and fecal samples were collected from DLs 21 to 22 and blood samples were collected on DL 22, via the inferior vena cava, from the maternal rats. The blood samples were centrifuged. The urine and fecal samples and the serum were shipped to the Sponsor. All Fo generation rats assigned to cross-fostering were sacrificed on DL 22 and a gross necropsy of the thoracic, abdominal and pelvic viscera was performed. The number and distribution of implantation sites were recorded. A liver section (right lateral lobe) from each dam was collected and shipped to the Sponsor. Rats that did not deliver a litter were sacrificed on DG 25 and examined for gross lesions. A blood sample was collected prior to sacrifice via the inferior vena cava; samples were centrifuged and serum samples were shipped to the Sponsor. A liver section (right lateral lobe) from each dam was retained and shipped to the Sponsor. Dams with no surviving pups were sacrificed after the last pup was found dead, missing or presumed cannibalized. A.2. F1 Generation Pups Assigned to Cross-Fostering: Day 1 of lactation (postpartum) was defined as the day of birth. Each litter was evaluated for viability at least twice each day of the postpartum period. The pups present in each litter were counted once each day. Physical signs were recorded for the pups once each day during the postpartum period. Pup body weights were recorded on DLs 1 (birth), 4, 7, 14 and 21 and at sacrifice. Pups found dead or sacrificed because of moribundity were examined for gross lesions and for the cause of death or the moribund condition. For all pups found dead on DLs 2 to 4, the lungs and the liver were preserved. Pups with gross 000 418-014:PAGE I-3 lesions found on DLs 2 to 4 were preserved and for pups found dead on DLs 5 to 21, the lungs, liver and gross lesions were preserved. On DL 4, cross-fostered F1 generation litters were culled to five male and five female pups per litter, where possible. Pups with gross lesions were preserved. The lungs and the liver were preserved. The lungs and the liver were collected from the first ten culled pups from each dosage group (irrespective of litter) from each dosage group on each day (control pool only on DL 1). The lungs and the liver were retained. Remaining culled pups were sacrificed and discarded without evaluation. On DL 21, all pups were sacrificed via decapitation and examined for gross lesions. Gross lesions were retained. Six litters per subset were randomly selected for collection of pooled samples (per litter) of blood and liver. Blood samples were collected via the inferior vena cava from each pup, pooled (per litter) and the samples were centrifuged. The serum samples were shipped to the Sponsor. A.3. Fo Generation Rats Assigned to Pharmacokinetic Sample Collection: Viability and clinical observations, body weights, feed consumption values and delivery observations were performed as previously described for Fo generation dams assigned to cross-fostering. Pharmacokinetic samples were collected from eight and two Fo generation rats in Groups I and II, respectively, on DL 14. Milk samples were collected approximately two to six hours postdosage on DL 14. The samples were shipped to the Sponsor. Following completion of milk sample collection, female rats were sacrificed and a gross necropsy of the thoracic, abdominal and pelvic viscera was performed. The number and distribution of implantation sites was recorded. Blood samples were collected via the inferior vena cava and centrifuged and the serum was shipped to the Sponsor. A liver section and the milk-secreting glands from the axillary, thoracic, abdominal and inguinal regions of each dam (left side only) were collected and shipped to the Sponsor. A.4. F1 Generation Pups Assigned to Pharmacokinetic Sample Collection: Viability and clinical observations, body weights and litter observations were performed as previously described for F1 generation pups assigned to crossfostering. On DL 14, litters assigned to the pharmacokinetic sample collection were sacrificed and blood samples were collected via the inferior vena cava from each pup, pooled (per litter), centrifuged and the serum was shipped to the Sponsor. The liver from each pup was collected, pooled (per litter), and shipped to the Sponsor. oooS B. Results All Fo generation female rats survived to scheduled sacrifice. All adverse clinical and necropsy observations were considered unrelated to the administration of PFOS. Body weight gains were reduced in the 1.6 mg/kg/day dosage group during the precohabitation and gestation periods, as compared to the control group values. Maternal body weight gains were variable throughout the lactation period and body weight gains for the entire lactation period were generally comparable among the four cross-fostered subsets. Absolute body weights were slightly reduced in the 1.6 mg/kg/day dosage group beginning at the end of the precohabitation period and continuing through the gestation and lactation periods. Absolute and relative feed consumption values were generally reduced for rats administered the 1.6 mg/kg/day dosage of PFOS during the precohabitation, gestation and lactation periods, as compared to the control group values. The duration of gestation tended to be reduced in the 1.6 mg/kg/day dosage group. As observed in a previously conducted study (Argus Research Laboratories, Inc., Protocol 418-008), this reduction was associated with minimal preimplantation loss; the average number of implantation sites per dam was slightly reduced, resulting in a slightly reduced delivered litter size. The duration of delivery per pup (in minutes) tended to be reduced in the 1.6 mg/kg/day dosage group. The live litter size before cross-fostering in the 1.6 mg/kg/day dosage group was also slightly reduced. The gestation index was 100% in each dosage group; all pregnant rats delivered live offspring. Pup mortality was greatest in pups from test article-treated dams fostered by test article-treated dams (subset C). Pup mortality was also increased in pups from test article-treated dams fostered by vehicle-treated dams (subset A). As a result of these increases in pup mortality, the viability indices (number of live pups on DL 4 preculling/number of pups cross-fostered DL1) were reduced, and numbers of surviving pups per litter and live litter sizes at weighing were reduced on DL 4 preculling in these two subsets. Pup mortality in pups from vehicletreated dams fostered by test article-treated dams (subset D) was comparable to that in pups from vehicle-treated dams fostered by vehicle-treated dams (subset B). These observations indicate that in tero exposure to the test article is responsible for pup mortality, and that potential exposure to the test article via maternal milk did not adversely affect the viability of the pups. Pup body weights were reduced on DL 1 in both subsets of dams administered PFOS (subsets C and D), as compared to pup body weights in the subsets of dams administered the vehicle (subsets A and B). Potential exposure to the test article via maternal milk after DL1 reduced pup body weights, regardless of in tero exposure to the 418-014:PAGE I-5 test article or vehicle. After DL 1, pup body weights were somewhat reduced in subsets D (pups from vehicle-treated dams fostered by test article-treated dams) and A (pups from test article-treated dams fostered by vehicle-treated dams), as compared to the pups from vehicle-treated dams cross fostered to vehicletreated dams (subset B). After DL 1, pup body weight reductions were greatest in subset C (pups from test article-treated dams fostered by test article-treated dams). Two litters in subset A (pups from test article-treated dams cross-fostered to vehicle-treated dams) and one litter in subset C (pups from test article-treated dams cross-fostered with test article-treated dams) had pups that did not nurse; pup mortality was greatest in these two subsets. There was no milk in the stomach of 57.1%, 100% and 87.0% of the pups that were found dead and necropsied in subsets A, C and D, respectively. Evaluation of the livers of pups that died or were sacrificed on DL 1 by electron microscopy revealed increased numbers of peroxisomes within hepatocytes of pups from dams treated with 1.6 mg/kg/day PFOS, compared with controls. Quantitation of peroxisomes indicated slightly more than twice the number of peroxisomes in treated pups. Differences in cellular membranes or mitochondria were not discernible between control and treated pups in the samples examined. Evaluation of the lungs of pups that died or were sacrificed on DL 1 with electron microscopy revealed Type II pneumocytes containing lamellar bodies in both control and 1.6 mg/kg/day dosage group pups, with an increased number of Type II pneumocytes and lamellar bodies in lungs from treated pups. Although some lamellar material (surfactant) was identifiable within alveolar lumina, no significant difference was noted between the control group and 1.6 mg/kg/day dosage group. 15* 418-014:PAGE I-6 C. Conclusion The 1.6 mg/kg/day dosage of PFOS caused reductions in body weight gain and reduced feed consumption values for the Fo generation dams, as occurred in a previous multigeneration study with the test article (Argus Research Laboratories, Inc., Protocol 418-008). As previously observed, the 1.6 mg/kg/day dosage of PFOS administered to the Fo generation dams in this study also reduced the durations of gestation and parturition, caused preimplantation loss and reductions in litter size and pup growth. Cross-fostering of pups from test article treated dams and vehicle treated dams revealed that the effects on pup mortality that occurred in the previously conducted study (418-008), can be attributed to in utero exposure to the test article, and that potential exposure to the test article via maternal milk did not adversely affect the viability of the pups. Exposure to the test article via both in utero exposure and maternal milk caused reductions in pup growth. Mildred S. Christian, Ph.D., Fellow, ATS Executive Director of Research Date Alan M. Hoberman, Ph.D., DABT Director of Research Date Study Director Z .l.:M L L z 3 .1 Date 25 e flti 418-014:PAGE 11-1 II. DESCRIPTION OF TEST PROCEDURES A. Conduct of Study: A.1. Sponsor: 3M Corporate Toxicology, 3M Center Building 220-2E-02, St. Paul, Minnesota 55144-1000 A.2. Testing Facility: Argus Research Laboratories, Inc., 905 Sheehy Drive, Building A, Horsham, Pennsylvania 19044-1297 A.3. Study Number: 418-014 A.4. Sponsor's Study Number: T-6295.13 A.5. Purpose of the Study: The purpose of this study was to evaluate survival of F1 generation pups following PFOS treatment of Crl:CDBR VAF/Plus Fo generation female rats during premating, gestation and lactation. F1 generation pups were cross-fostered during the lactation period to differentiate between effects on pups exposed to the test article in utero and pups exposed to the test article via maternal milk. Selected pharmacokinetic samples were collected from Fo generation female rats and F1 generation pups. A.6. Study Design: The requirements of the U.S. Food and Drug Administration (FDA)<1)were used as the basis of study design. A.7. Regulatory Compliance: The study was conducted in compliance with Good Laboratory Practice (GLP) regulations of the U.S. Food and Drug Administration (FDA)(2), the Japanese Ministry of Health and Welfare (MHW)<3) and the European Economic Community (EEC)<4). There were no deviations from the GLP regulations that affected the quality or integrity of the study. Quality Assurance Unit findings derived from the 160 000 418-014:PAGE II-2 inspections during the conduct of this study are documented and have been provided to the Study Director and the Testing Facility Management. A.8. Ownership of the Study: The Sponsor owns the study. All raw data, analyses, reports and preserved tissues are the property of the Sponsor. A.9. Study Monitor: Marvin T. Case, D.V.M., Ph.D. A.10. Alternate Study Monitor: Andrew M. Seacat, Ph.D. A.11. Study Director: Raymond G. York, Ph.D., DABT (Associate Director of Research) A.12. Technical Performance: John F. Barnett, B.S. (Director of Laboratory Operations) Margaret M. Martin (Research Associate) Corrie L. Pabst (Quality Control Associate) A.13. Report Preparation: Raymond G. York, Ph.D., DABT Jo Ann Frazee, M.S. (Study Coordinator) Susan K. Bradshaw, B.S. (Data Management Specialist) Karen G. Parker, A.A. (Report Administrator) A.14. Report Review: Alan M. Hoberman, Ph.D., DABT (Director of Research) Mildred S. Christian, Ph.D., Fellow, ATS (Executive Director of Research) A.15. Date Protocol Signed: 23 October 1998 418-014:PAGE II-3 A. 16. Dates of Technical Performance: Rat Arrival 27 OCT 98 Dosage Period (42 days before cohabitation until DLa21) 02 NOV 98 - 28 JAN 99 Cohabitation Period 13 DEC 98 PM - 19 DEC 98 AM Day 0 of Presumed Gestation (DG 0) 14 DEC 9 8 -1 9 DEC 98 Delivery Period 04 JAN 99 - 09 JAN 99 DL 1 Culling (Control Pool) 04 JAN 99 - 09 JAN 99 DL 4 Culling (Cross-Fostered Litters) 08 JAN 99-11 JAN 99 DG 25 Sacrifice 08 JAN 9 9 -1 2 JAN 99 DL 14 Pharmacokinetic Group Sacrifice 17 JAN 9 9 -2 2 JAN 99 F1 Generation Pups Sacrifice (DL 21) 25 JAN 99 - 28 JAN 99 Fo Generation Dams Sacrifice (DL 22) 26 JAN 99 - 29 JAN 99 A. 17. Records Maintained: The original report, raw data and reserve samples of the bulk test article and vehicle components are retained in the archives of Argus Research Laboratories, Inc. Any preserved tissues are retained in the archives of the Testing Facility for one year after the mailing of the draft final report, after which the Sponsor will decide their final disposition. All unused prepared test article formulations were discarded at the Testing Facility. Unused bulk test article was returned to the Study Monitor. B. Test Article Information: B.1. Description: PFOS (FC-95) - a light-colored powder B.2. Lot/Batch Number: 217 (Expiration date: May 2000) B.3. Date Received and Storage Conditions: The test article was received on 21 October 1998, and stored at room temperature. a. DL is used as an abbreviation for day of lactation or day postpartum. 418-014:PAGE II-4 B.4. Special Handling Instructions: Standard safety precautions (use of protective clothing, gloves, dust-mist respirator and safety goggles) were taken when handling the bulk test article and prepared formulations. B. 5. Analysis of Activity: Information regarding the identity, strength, composition and purity of the test article is on file with the Sponsor. C. Vehicle Information: C.1. Description: 0.5% Tween 80 in Reverse Osmosis Membrane Processed Deionized Water (R.O. Deionized Water) C.2. Lot Numbers: Tween 80 - M03H05, L06662 and M29477 C.3. Dates Received and Storage Conditions: The Tween 80 was received on 3 December 1998 (lot M03H05), 1 September 1998 (lot L06662) and 17 September 1998 (lot M29477), from J.T. Baker, Phillipsburg, New Jersey, and was stored at room temperature. The R.O. deionized water is available from a continuous source at the Testing Facility and is maintained at room temperature. C.4. Special Handling Instructions: Standard safety precautions (use of protective clothing, gloves, dust-mist respirator, safety goggles or safety glasses and a face-shield) were taken when handling the vehicle. C.5. Analysis of Purity: Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to be present in the vehicle that would interfere with the results of this study. 418-014:PAGE M-5 D. Test Article Preparation and Storage Conditions: Suspensions were prepared daily at the Testing Facility at concentrations of 0 and 0.32 mg/mL. Records are maintained in the raw data to document how the test article formulations were prepared. Prepared suspensions were stored at room temperature. D.1. Sample information: Sample Type Concentration (all levels) Test Article Reserve Vehicle Components Reserve R.O. Deionized Water Tween 80 Lot L06662 Lot M29477 Lot M03H05 Size 2 mL* 19 Date Retained 01 NOV 98 27 JAN 99 28 OCT 98 5 mL 5 mL 5 mL 5 mL 28 OCT 98 28 OCT 98 10 NOV 98 09 DEC 98 Shipping/Storage Conditions Frozen on dry ice Room temperature Shipped To Sponsor Testing Facility Archives Date Shipped 02 NOV 98 27 JAN 99 25 NOV 98 Room temperature Testing Facility Archives 25 NOV 98 25 NOV 98 25 NOV 98 27 JAN 99 a. Duplicate samples were taken from the first and last preparation on the day prepared. One set of samples was sent for analysis; the remaining samples were retained at the Testing Facility as backups. Backup samples were stored (-70C or below) and will be discarded at the Testing Facility upon request of the Sponsor. D. 2. Analytical Results: Data verifying the stability of the test article in the vehicle for 48 hours under the conditions of administration are on file with the Sponsor. Homogeneity of the prepared formulations is on file with the Sponsor. Results of the concentration analyses have not been received. E. Test System: E.1. Species: Rat E.2. Strain: Crl:CDBR VAF/Plus (Sprague-Dawley) 418-014:PAGE II-6 E.3. Supplier (Source): Charles River Laboratories, Inc., Portgage, Michigan E.4. Sex: Female (Male rats of the same source and strain were used only as breeders and were not considered part of the Test System). E.5. Rationale for Test System: The Crl:CDBR VAF/Plus (Sprague-Dawley) rat was selected as the Test System because: 1) this strain of rat was used in the reproductive and developmental toxicity studies; 2) historical data and experience exist at the Testing Facility5'7'; and 3) the test article is pharmacologically active in the species and strain. E.6. Test System Data: Number of Rats Approximate Date of Birth Approximate Age at Arrival Weight (g) on the Day after Arrival Weight (g) at Study Assignment 86 23 AUG 98 66 days 179 - 226 200 - 244 E.7. Breeder Male Rat Data: Number of Rats Approximate Date of Birth Approximate Age at Arrival Weight (g) on the Day After Arrival Weight (g) at Cohabitation 110 13 JAN 98 77 days 317 - 352 514 - 866 E.8. Method of Randomization: Upon arrival, male and female rats were assigned to individual housing on the basis of computer-generated random units. After acclimation, 78 virgin female rats were selected for study on the basis of physical appearance and body weights recorded during acclimation. The female rats were assigned to dosage groups (42 in the control group and 36 in the treated group) based on computer generated (weight-ordered) randomization procedures. Within each dosage group, consecutive order was used to assign female rats to cohabitation with breeder male rats, one male rat per female rat. Rats were assigned to cross-fostering or pharmacokinetic collection depending on the order 418-014:PAGE II-7 of delivery and the availability of rats for cross-fostering. Eight female rats from the control group and two female rats from the treated group were designated for pharmacokinetic sample collection following delivery; these rats were not crossfostered3. Twenty-five litters per dosage group were cross-fostered on DL 1b. The litters were assigned such that four subsets were obtained among the two dosage groups, as described in the table below. Subset assignments were made based on order of deliveries. Dosage Group I I II II Number of Mated Fem ale Rats 13 12 12 13 Dosage (m g/kg/day) 0 (Vehicle) 0 (Vehicle) 1.6 1.6 Number of Litters R e a s s ig n e d 13 12 12 13 R eassigned Litter Type (Dam Treatment) Treated Untreated Treated Untreated Subset Number A B C D On DL 4, cross-fostered litters were culled to five male and five female pups per litter, where possible, using a table of random units. E.9. System of Identification: E.9.a. Fo Generation Rats: Rats were permanently identified using Monel self-piercing ear tag (Gey Band and Tag Co., Inc., No. MSPT 20101). Male rats were given unique permanent identification numbers upon assignment to the Testing Facility's breeder male rat population. Female rats were assigned temporary numbers at receipt and given unique permanent identification numbers when assigned to the study. Cage tags were marked with the study number, permanent rat number, sex, test article identification and dosage level. E.9.b. F1 Generation Pups: Pups were not individually identified during lactation; all parameters were evaluated in terms of the litter. a. See APPENDIX D (DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY), items 1 and 2. b. See APPENDIX D, item 3. 418-014:PAGE II-8 F. Husbandry: F.1. Research Facility Registration: USDA Registration No. 23-R-099 under the Animal Welfare Act, 7 U.S.C. 2131 et seq. F.2. Study Rooms: The study rooms were maintained under conditions of positive airflow relative to a hallway and independently supplied with a minimum often changes per hour of 100% fresh air that had been passed through 99.97% HEPA filters. Room temperature and humidity were monitored constantly throughout the study. Room temperature was targeted at 64F to 79F (18C to 26C); room humidity was targeted at 30% to 70%. See APPENDIX E (TEMPERATURE AND RELATIVE HUMIDITY REPORTS). F.3. Housing: Fo generation rats were individually housed in stainless steel, wire-bottomed cages, except during the cohabitation and postpartum periods. During the cohabitation period, each pair of rats was housed in the male rat's cage. Beginning no later than DG 20, Fo generation female rats were individually housed in nesting boxes, except during the collection interval for urine and fecal samples. During this collection interval (DLs 21 to 22), the female rats were housed individually in metabolism cages. Each dam and assigned litter were housed in a common nesting box during the postpartum period. All cage sizes and housing conditions were in compliance with the Guide for the Care and Use of Laboratory Animals<8). F.4. Lighting: An automatically-controlled fluorescent light cycle was maintained at 12-hours light: 12-hours dark, with each dark period beginning at 1900 hours EST. F.5. Sanitization: Cage pan liners were changed approximately three times each week. Cages were changed approximately every other week. Bedding was changed as often as necessary to keep the rats dry and clean. F.6. Feed: Rats were given ad libitum access to Certified Rodent Diet #5002 (PMI Nutrition International, St. Louis, Missouri) in individual feeders. QflOgSO ouy 418-014:PAGE II-9 F.7. Feed Analysis: Analyses were routinely performed by the feed supplier. No contaminants at levels exceeding the maximum concentration limits for certified feed or deviations from expected nutritional requirements were detected by these analyses. Copies of the results of the feed analyses are available in the raw data. Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the feed that would have interfered with the results of this study. F.8. Water: Local water that had been processed by passage through a reverse osmosis membrane (R.O. water) was available to the rats ad libitum from an automatic watering access system and/or individual water bottles. Chlorine was added to the processed water as a bacteriostat. F.9. Water Analysis: The processed water is analyzed twice annually for possible chemical contamination (Lancaster Laboratories, Lancaster, Pennsylvania) and monthly for possible bacterial contamination (Analytical Laboratories, Inc., Chalfont, Pennsylvania). Copies of the results of the water analyses are available in the raw data. Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the water that would have interfered with the results of this study. F.10. Bedding: Bed o'cobs was used as the nesting material (Andersons' Industrial Products Groups, Maumee, Ohio). F.11. Bedding Analysis: Neither the Sponsor nor the Study Director was aware of any potential contaminants likely to have been present in the bedding that would have interfered with the results of this study. Analyses for possible contamination are conducted annually and documented in the raw data. 0 0 ( 1234. G. Methods: G.1. Dosage Administration: 418-014:PAGE IMO Dosage Group I II Dosage (mg/kg/day)* Concentration (mg/mL) Dosage Volume (mL/kg) 0 (Vehicle) 1.6 0 0.32 5 5 a. The test article was considered 100% pure for the purpose of dosage calculations. Dosage Group I Rats Dosed Until Cohabitation Number of Rats Assigned Numbers 42 18901 - 18942 II 36 18943- 18978 Rats Assigned to Pharmacokinetic Evaluation Number Assigned Numbers of Rats 18910, 18911, 18913, 8 18915, 18926, 18927, 18939, 18940 2 18956, 18971 Dosage Subset Group Number Assigned Dam and Utter Numbers* Dams Litters 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. 18942 18965 I 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 II C 18943, 18947, 18948, 18957, 18960, 18964, 18968, 18973, 18970, 18969, 18974, 18967, 18967, 18974, 18969, 18970, 18973, 18968 18964, 18960, 18957, 18948, 18947, 18943 II 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, 18965 18942 a. Dams and their cross-fostered litters are listed in consecutive order. G.2. Rationale for Dosage Selection: Dosages were selected on the basis of a previous two-generation study conducted with the test article (Argus Research Laboratories, Inc., Protocol 418-008). In this study the Fo generation maternal and paternal noobservable-effect-level (NOEL) of PFOS was 0.1 mg/kg/day (0.4 mg/kg/day and higher dosages caused reductions in body weight gain and reduced feed consumption values). The Fo generation reproductive NOEL was greater than 3.2 mg/kg/day; no effects on mating, fertility or estrous cycling occurred. The NOEL for viability and growth in the F1 generation offspring was 0.4 mg/kg/day (1.6 mg/kg/day and higher dosages caused preimplantation loss and reductions in litter size, pup viability, growth and survival). 418-014:PAGE 11-11 The F1 generation maternal and paternal NOEL of PFOS was 0.1 mg/kg/day (0.4 mg/kg/day dosage caused reductions in body weight gain and reduced feed consumption values). The F1 generation reproductive NOEL was greater than a dosage of 0.4 mg/kg/day; no effects on mating or fertility occurred. The NOEL for viability and growth in the F2 generation offspring was 0.1 mg/kg/day (0.4 mg/kg/day dosage caused stillbirths and reductions in litter size, pup viability, growth and survival). G.3. Route of Administration: Oral (gavage) G.4. Rationale for Route of Administration: The oral (gavage) route was selected for use because: 1) this was the route of administration in the developmental and reproductive toxicology studies; and 2) it is one of the possible routes of human exposure. G.5. Frequency of Administration: G.5.a. Fo Generation Female Rats: Female rats were given the test article or the vehicle once daily beginning 42 days prior to cohabitation through DL 21. Dosages were adjusted daily for body weight changes and given at approximately the same time each day. G.5.b. F1 Generation Pups: F1 generation pups were not directly given the test article, but may have been possibly exposed to the test article during maternal gestation (in utero exposure) or via maternal milk during the lactation period. G.6. Length of Study: Approximately 3 months G.7. Method of Study Performance: G.7.a. Cohabitation and Cross-Fostering Procedures: Twenty-five rats assigned to the control group were cohabited one day prior to cohabitation for rats assigned to the treated group and the cohabitation period consisted of a maximum of six days3. The cohabitation period for the remaining a. See APPENDIX D, item 4. 418-014:PAGE 11-12 rats in the control group and the rats assigned to the treated group was a maximum of five days. During cohabitation, both control and PFOS-treated groups were co-housed with untreated male breeders, one male rat per one female rat. Female rats with spermatozoa observed in a smear of the vaginal contents and/or a copulatory plug observed in situ were considered to be at DG 0 and assigned to individual housing. Several dams assigned to the control group that delivered on the first day of parturition occurred were allowed to deliver and retain their natural pups until needed for cross-fostering purposes. This provided a pool of control dams available for immediate cross-fostering of pups from PFOS-treated dams, thus preventing these pups from nursing from their PFOS-treated dams. Starting with the deliveries of the dams on the second day, all pups were removed immediately (as soon as parturition was completed) from their respective dams and placed with either another dam assigned to the control group or a PFOS-treated dam. When a dam assigned to the control group was not available, then a dam from the control pool was used. The litter from that control pool dam was then sacrificed via decapitation. This cross-fostering procedure resulted in four groups of 12 or 13 dams/pups, i.e., Control/PFOS (Subset A), Control/Control (Subset B), PFOS/PFOS (Subset C) and PFOS/Control (Subset D). This procedure allowed distinctions to be made between prenatal and postnatal effects of the continuous maternal PFOS treatment. G.7.b. Fo Generation Rats Assigned to Cross-Fostering: The Fo generation female rats were observed for viability at least twice each day of the study and for general appearance at least once during the acclimation period. The rats were also examined for clinical observations of effects of the test article, abortions, premature deliveries and/or deaths before and approximately one hour postdosage. Body weights were recorded at least once during the acclimation period. Body weights were recorded daily during the dosage period and at sacrifice. Feed consumption values were recorded at least once during the acclimation period, weekly to cohabitation, daily during the presumed gestation period and on DLs 4, 7, 10 and 14. Feed consumption was not tabulated after DL 14, when it was expected that the pups began to consume maternal feed. The female rats were continually evaluated (24 hours per day) for clinical observations during parturition [performed under red light conditions during the dark period (time each pup was observed was recorded)], duration of gestation (DG 0 to the time the first pup was delivered), length of partuition (time of oooet 1T| IMP 418-014:PAGE 11-13 observation of last pup minus the time of observation of the first pup divided by N-1 pups in each litter), litter sizes (defined as all pups delivered) and pup viability at birth. Pups that either appeared stillborn or that died before initial examination of the litter for viability were examined for vital status at birth. The lungs were removed and immersed in water. Pups with lungs that sank were considered stillborn; pups with lungs that floated were considered liveborn and to have died shortly after birth. Maternal behavior of the dams was evaluated daily when the pups were examined during the 21-day postpartum period. Observed maternal behavior was recorded on DLs 1,4, 7, 14 and 21. Deviations from expected maternal behavior were recorded, if and when present, on all other days during the postpartum period. Fo generation female rats were housed individually in metabolism cages for collection of urine and fecal samples from DLs 21 to 22. Following the 24-hour collection interval, samples were collected into centrifuge tubes, placed on dry ice, stored frozen (-70C or below) and shipped to the Sponsor for analysis. On DL 22, blood samples (approximately 4 mL each) were collected from the maternal rats via the inferior vena cava and transferred into serum separator tubes after removal from metabolism caging. The samples were spun in a refrigerated centrifuge. The serum was transferred into polypropylene tubes labeled with the study number, rat identification, date of collection, study day and collection timepoint. All samples were immediately frozen on dry ice, stored frozen (-70C or below) and shipped to the Sponsor for analysis. G.7.c. F1 Generation Pups Assigned to Cross-Fostering: Day 1 of lactation (postpartum) was defined as the day of birth and was also the first day on which all pups in a litter were individually weighed. Each litter was evaluated for viability at least twice each day of the postpartum period. Dead pups observed at these times were removed from the nesting box. The pups present in each litter were counted once each day. Physical signs (including gross external physical anomalies) were recorded for the pups once each day during the postpartum period. Pup body weights were recorded on DLs 1 (birth), 4, 7, 14 and 21 and at sacrifice. On DL 4, cross-fostered F1 generation litters were culled to five male and five female pups per litter, where possible. m t if f 418-014:PAGE 11-14 G.7.d. Fo Generation Rats Assigned to Pharmacokinetic Sample Collection: Viability and clinical observations, body weights, feed consumption values and delivery observations were performed, as previously described for Fo generation dams assigned to cross-fostering. Blood, milk and liver samples were collected from eight and two Fo generation rats in Groups I and II, respectively, designated for pharmacokinetic sample collection per dosage group on DL 14. Milk samples were collected approximately two to six hours postdosage on DL 14. Each dam was removed from the nesting box and individually housed for approximately four hours. The dam was injected intravenously with one unit of oxytocin (20 USP units/mL, Lot Number 52064, expiration date July, 2000, manufactured by Osborn) approximately five minutes before milk samples (at least 100 pL per sample) were collected. The samples were immediately frozen on dry ice, stored frozen (-70C or below) and shipped to the Sponsor. Following milk sample collection, blood samples (approximately 4 mL each) were collected from the maternal rats via the inferior vena cava and transferred into serum separator tubes. The samples were spun in a refrigerated centrifuge. The serum was transferred into polypropylene tubes labeled with the study number, rat identification, date of collection, study day and collection timepoint. All samples were immediately frozen on dry ice, stored frozen (-70C or below) and shipped to the Sponsor. Following collection of milk and blood samples, a liver section (right lateral lobe) and the milk-secreting glands from the axillary, thoracic, abdominal and inguinal regions of each dam (left side only) were collected, stored frozen (-70C or below) and shipped to the Sponsor. G.7.e. F1 Generation Pups Assigned to Pharmacokinetic Sample Collection: Viability and clinical observations, body weight and litter observations were performed as previously described for F1 generaton pups assigned to crossfostering. Caps and labeled tubes were weighed (combined weight, to the nearest 0.001 g) before and after retention of pooled pup samples for subsequent use in pharmacokinetic analyses. Blood samples were collected via the inferior vena cava from each pup on DL 14, pooled (per litter) and transferred into serum separator tubes. The samples were spun in a refrigerated centrifuge. The serum was transferred into polypropylene tubes labeled with the study number, pup identification, date of t il* m 418-014:PAGE 11-15 collection, study day and collection timepoint. All samples were immediately frozen on dry ice, stored frozen (-70C or below) and shipped to the Sponsor for analysis. The liver from each pup was collected, pooled (per litter), stored frozen (-70C or below) and shipped to the Sponsor for analysis. H. Gross Necropsy3: All Fo generation rats were sacrificed by carbon dioxide asphyxiation, and a gross necropsy of the thoracic, abdominal and pelvic viscera was performed. Gross lesions were retained in neutral buffered 10% formalin for possible future evaluation. Representative photographs of maternal gross lesions are available in the raw data. H.1. Fo Generation Rats: The female rats were sacrificed on DL 22. The number and distribution of implantation sites were recorded. A liver section (right lateral lobe) from each dam was collected, stored frozen (-70C or below) and shipped to the Sponsor for analysis. Following completion of milk sample collection on DL 14, female rats assigned to pharmacokinetic collection were sacrificed. Blood and liver samples were collected as previously described. The number and distribution of implantation sites was recorded. Rats that did not deliver a litter were sacrificed on DG 25 and examined for gross lesions. Uteri were stained with 10% ammonium sulfide to confirm the absence of implantation sites<9>. Dams with no surviving pups were sacrificed after the last pup was found dead, missing or presumed cannibalized. A blood sample (approximately 4 mL) was collected, prior to sacrifice, via the inferior vena cava and transferred into serum separator tubes. The samples were spun in a refrigerated centrifuge. The serum was transferred into a polypropylene tube labeled with study number, rat identification, date of collection, study day and collection timepoint. The samples were immediately frozen on dry ice, and stored frozen (-70C or below) and a. A table of random units was used to select one Fo generation control group female rat and one F1 generation control group male and female pup from which all tissues examined at necropsy were retained, in order to provide control tissues for potential comparative histopathological evaluations. 418-014:PAGE 11-16 shipped to the Sponsor for analysis. A liver section (right lateral lobe) from each dam was collected, stored frozen (-70C or below) and shipped to the Sponsor for analysis. H.2. F1 Generation Pups: Pups that died before examination of the litter for pup viability were evaluated for vital status at birth, as previously described. Pups with gross lesions were preserved in Bouin's solution for possible future evaluation. The lungs were preserved in Bouin's solution, and the liver was preserved in neutral buffered 10% formalin for possible future evaluation. Pups from dams assigned to the control pool and culled on DL 1 and all other pups culled on DL 4 were sacrificed via decapitation. The lungs and the liver were collected from the first ten culled pups from the control pool (irrespective of litter) and the first ten pups that were found dead (no autolysis) from each dosage group on DL 1. The lungs were retained in Bouin's solution, and the livers were retained in neutral buffered 10% formalin for possible future evaluation. Remaining culled pups were sacrificed and discarded without evaluation. Pups found dead or sacrificed because of moribundity were examined for gross lesions and for the cause of death or the moribund condition. For all pups found dead on DLs 2 to 4, the lungs were preserved in Bouin's solution, and the liver was preserved in neutral buffered 10% formalin, for possible future evaluation. Pups with gross lesions found on DLs 2 to 4 were preserved in Bouin's solution for possible future evaluation. For all pups found dead on DLs 5 to 21, the lungs, liver and gross lesions were preserved in neutral buffered 10% formalin for possible future evaluation. On DL 14, litters assigned to the pharmacokinetic sample collection and their respective dams were sacrificed by carbon dioxide asphyxiation. Individual, pooled samples (per litter) of blood and liver were collected as previously described. The pups were examined for gross lesions. On DL 21, all pups were sacrificed via decapitation and examined for gross lesions. Gross lesions were retained in neutral buffered 10% formalin. Six litters per subset were randomly selected for collection of pooled samples (per litter) of blood and liver. Blood samples were collected via the inferior vena cava from each pup, pooled (per litter) and transferred into serum separator tubes. The samples were spun in a refrigerated centrifuge. The serum was transferred to polypropylene tubes labeled with the study number, rat identification, date of collection, study day and collection timepoint. All samples were immediately frozen on dry ice, stored frozen (-70C or below) and shipped to the Sponsor for analysis. n A iy M f t M ill 418-014:PAGE 11-17 All pups selected for continued observation were sacrificed via decapitation and examined for gross lesions. I. Statistical Analyses: Averages and percentages were calculated. Litter values were used where appropriate. m III. RESULTS 418-014:PAGE 111-1 A. Mortality, Clinical and Necropsy Observations (Summaries - Tables 1 and 2: Individual Data - Tables 19 and 20) All rats survived to scheduled sacrifice. All adverse clinical observations during the precohabitation, gestation and lactation periods were considered unrelated to the test article because the incidences were not dosage-dependent and/or the observations are commonly seen in rats in the laboratory environment. These observations included chromorhinorrhea, scaly tail, abrasion on the head, neck, tail and/or forelimb, missing, broken and/or misaligned incisors, localized alopecia on the head, limbs, neck and/or back, chromodacryorrhea, scab on the head or tail, dehydration, perineal hernia, broken or swollen forepaw digit and protruding xiphoid. The only necropsy observation was an inguinal mass (presumed to be a galactocele) in one control group dam assigned to pharmacokinetic sample collection on lactation day 14 (DL 14). B. Body Weights (Figures 1 and 2; Summaries - Tables 3 through 8: Individual Data - Tables 21 through 23) Body weight gains during the precohabitation period were reduced as compared to the control group values, beginning on days 15 to 22 of the study (DSs 15 to 2) and continuing through DSs 29 to 36. Rats in both dosage groups lost weight on DSs 36 to 43; the body weight loss was greater in the 1.6 mg/kg/day dosage group. Although calculated for a seven-day interval, the weight losses generally occurred on days 42 to 43 of the study and were associated with movement of the male breeder rats into the animal room and mating activity in the animal room when the rats were cohabited on day 42. As a result of these changes, body weight gains for the entire precohabitation period (DSs 1 to 43) were reduced and absolute body weights were slightly reduced on DS 43 (96.6% of control) in the 1.6 mg/kg/day dosage group. Maternal body weight gains continued to be reduced in the 1.6 mg/kg/day dosage group in the first two weeks of the gestation period (days 0 to 14 of gestation, DGs 0 to 14). As a result, body weight gains were reduced for the entire gestation period (DGs 0 to 20) in the 1.6 mg/kg/day dosage group. Maternal body weights tended to be reduced in the 1.6 mg/kg/day dosage group throughout gestation. Reduced maternal body weights persisted in the two subsets of rats administered 1.6 mg/kg/day PFOS during the lactation period (subsets C and D), ---- trap 418-014:PAGE III-2 as compared to the rats in the subsets administered the vehicle (subsets A and B). Lactation body weights had the expected degree of variability but were comparable between subsets A and B (vehicle control groups) and between subsets C and D (test article-treated groups). Maternal body weight gains for the entire lactation period (DLs 1 to 22) were essentially comparable among the four subsets. C. Absolute q/davl and Relative q/kq/dav) Feed Consumption Values Summaries - Tables 9 through 14: Individual Data - Tables 24 through 26) Absolute (g/day) and relative (g/kg/day) feed consumption values were generally reduced for rats administered the 1.6 mg/kg/day dosage of PFOS during the precohabitation, gestation and lactation periods, as compared to the control group values. D. Natural Delivery and Litter Observations Summaries - Tables 15 and 16: Individual Data - Tables 27 through 31) D.1. Natural Delivery Observations The duration of gestation tended to be reduced in the 1.6 mg/kg/day dosage group, both when the duration of gestation was calculated in whole days and to the nearest tenth of a day. As previously observed, this reduction was associated with minimal preimplantation loss; the average number of implantation sites per dam was slightly reduced, resulting in a slightly reduced delivered litter size. The duration of delivery per pup (in minutes) tended to be reduced in the 1.6 mg/kg/day dosage group. The live litter size before crossfostering in the 1.6 mg/kg/day dosage group was also slightly reduced, an observation related to total litter size and associated with stillbirths. The gestation index was 100% in each dosage group; all pregnant rats delivered live offspring. D.2. Cross-Fostered Litter Observations After cross-fostering on DL 1, live litter sizes were comparable among the four subsets. Pup mortality was greatest in subset C (pups from test article-treated dams fostered by test article-treated dams); 19.2% of the pups were found dead or presumed cannibalized on DLs 2 to 4. Pup mortality was also increased in subset A (pups from test article-treated dams fostered by vehicle-treated dams) on DLs 2 to 4; 9.0% of these pups were found dead or presumed cannibalized. As a result of these increases in pup mortality, the viability indices (number of W 418-014:PAGE III-3 live pups on DL 4 preculling/number of pups cross-fostered on DL1) were reduced, and numbers of surviving pups per litter and live litter sizes at weighing were reduced on DL 4 preculling in these two subsets. Pup mortality in subset D (pups from vehicle-treated dams fostered by test article-treated dams) was comparable to that in subset B (pups from vehicle-treated dams fostered by vehicle-treated dams). These observations indicate that in tero exposure to the test article is responsible for pup mortality, and that potential exposure to the test article via maternal milk did not adversely affect the viability of the pups. Pup body weight/litter were reduced on DL 1 in both subsets of pups from dams administered PFOS (subsets A and C), as compared to pup body weights in the subsets of pups from dams administered the vehicle (subsets B and D). Potential exposure to the test article via maternal milk after DL1 reduced pup body weights, regardless of in tero exposure to the test article or vehicle. After DL 1, pup body weights were somewhat reduced in subsets D (pups from vehicle-treated dams fostered by test article-treated dams) and A (pups from test article-treated dams fostered by vehicle-treated dams), as compared to the pups from vehicle-treated dams fostered by vehicle-treated dams (subset B). After DL 1, pup body weight reductions were greatest in subset C (pups from test article-treated dams fostered by test article-treated dams). Sex ratios and the lactation index (number of live pups on DL 21/number of live pups on DL 4 postculling) were comparable among the four cross-fostered subsets. E. Clinical Observations from Birth to Dav 21 Postpartum and Necropsy Observations (Summaries - Tables 17 and 18: Individual Data Tables 32 and 33) Two litters in subset A (pups from test article-treated dams fostered by vehicletreated dams) and one litter in subset C (pups from test article-treated dams fostered by test article-treated dams) had pups that did not nurse; pup mortality was greatest in these two subsets. There was no milk in the stomach of 57.1%, 100% and 87.0% of the pups that were found dead and necropsied in subsets A, C and D, respectively. All other clinical and necropsy observations in the F1 generation pups were considered unrelated to the test article because: 1) the incidences were not dosage-dependent; or 2) the observation occurred in only one litter. Clinical observations included black tip of tail, labored breathing, dark in color, missing hindpaw digit and bent tail. One pup in subset A had slight dilation of the renal pelvis at necropsy on DL 21. wtnzsz* mv 418-014:PAGE III-4 F. Electron Microscopic Evaluation of Liver and Luna from Dav 1 Postpartum Pups (APPENDIX G1 F.1. Liver Ultrastructurally, there were increased numbers of peroxisomes within hepatocytes of pups from dams treated with 1.6 mg/kg/day PFOS, compared with controls. Quantitation of peroxisomes indicated slightly more than twice the number of peroxisomes in treated pups. Differences in cellular membranes or mitochondria were not discernible between control and treated pups in the samples examined. F.2. Lung Type II pneumocytes containing lamellar bodies were identified in both control and 1.6 mg/kg/day dosage group pups; however, there appeared to be an increased number of Type II pneumocytes and lamellar bodies in lungs from treated pups. Although some lamellar material (surfactant) was identifiable within alveolar lumina, no significant difference was noted between the control group and 1.6 mg/kg/day dosage group. 7% > m m REFERENCES 418-014:PAGE III-5 1. U.S. Food and Drug Administration (1994). International Conference on Harmonisation; Guideline on detection of toxicity to reproduction for medicinal products. Federal Register, September 22, 1994, Voi. 59, No. 183. 2. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. 3. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. 4. European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles of good laboratory practice. Official Journal of the European Communities: Legislation. 32 (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. National Technical Information Service, U.S. Department of Commerce, Springfield, VA 22161. 6. Christian, M.S. (1984). Reproductive toxicity and teratology evaluations of naltrexone (Proceedings of Naltrexone Symposium, 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:CDBR Rat. Charles River Laboratories, Inc., Wilmington, MA 01887-0630. (Data base provided by Argus Research Laboratories, Inc.) 8. Institute of Laboratory Animal Resources (1996). Guide for the Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 9. Salewski, E. (1964). Farbemethode zum makroskopischen Nachweis von Implantationsstellen am Uterus der Ratte. Arch. Pathol. Exp. Pharmakol. 247:367. APPENDIX A REPORT FIGURES o *i MATERNAL BODY WEIGHTS Figure 1 W E IG H T (G) 418-014: PAGE A-1 iMsooe 380 ; 370 i360 i -iI S -350 - j I MATERNAL BODY WEIGHTS RATS ASSIGNED TO CROSS-FOSTERING Figure 2 330 - 320 - ! -ij 310 --- - T .. .. 1 ) 10 DAY OF LACTATION 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-fostered with vehicle control group liners. l8 14 2I2 SUBSET A a SUBSET B b -o- SUBSETCc SUBSET Dd 418-014: PAGE A-2 APPENDIX B REPORT TABLES PROTOCOL 416-014: 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 DOSAGE (MG/KG/DAY) I 0 (VEHICLE) PRECOHABITATION (DAY 1 TO 42 OF STUDY):a MAXIMUM POSSIBLE INCIDENCE 1764/ 42 MORTALITY 0 CHROMORHINORRH EA 1/ 1 SCALY TAIL 0/ 0 ABRASION b 23/ 2 LOCALIZED ALOPECIA: TOTAL HEAD LIMBS NECK BACK 104/ 49/ 57/ 32/ 11/ 5 3 2 1 1 INCISORS: TOTAL MISALIGNED MISSING/BROKEN 16/ 3 14/ 2 2/ 1 CHROMODACRYORRHEA 5/ 1 HEAD: SCAB 2/ 1 MAXIMUM POSSIBLE INCIDENCE = (DAYS X RATS)/NUMBER OF RATS EXAMINED PER GROUP. N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. a. Day 42 of study was the first day of cohabitation for rats assigned from Group b. Located on the head, neck, tail or left forelimb. I. II 1.6 1512/ 36 0 3/ 2 35/ 1 7/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 418-014:PAGE B-1 PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 1 (PAGE 2) : CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) PRESUMED GESTATION: a MAXIMUM POSSIBLE INCIDENCE 896/ 40 MORTALITY 0 LOCALIZED ALOPECIA: TOTAL LIMBS NECK BACK UNDERSIDE 21/ 2 13/ 1 0/ 0 8/ 1 0/ 0 SCALY TAIL 0/ 0 INCISORS: MISALIGNED 39/ 2 CHROMORHINORRHEA 4/ 2 CHROMODACRYORRHEA 15/ 1 DEHYDRATION 1/ 1 PERINEAL HERNIA 1/ 1 MAXIMUM POSSIBLE INCIDENCE = (DAYS X RATS)/NUMBER OF RATS EXAMINED PER GROUP. N/N = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION, a. Restricted to rats with a confirmed mating date. II 1.6 752/ 34 0 50/ 4 32/ 3 18/ 1 0/ 0 8/ 1 21/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 418-014:PAGE B-2 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 1 (PAGE 3): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS RATS ASSIGNED TO CROSS-FOSTERING SUBSET DOSAGE GROUP DOSAGE (MG/KG/DAY) A Ia 0 (VEHICLE) B Ib 0 (VEHICLE) c II c 1.6 LACTATION : MAXIMUM POSSIBLE INCIDENCE 286/ 13 264/ 12 246/ 12 MORTALITY 000 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 16/ 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. N/N = 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 grou p dams cross-fostered with 1.6 mg/kg/day dosage gr ou p litters. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. D II d 1.6 267/ 13 0 22/ 1 22/ 1 7/ 1 3/ 1 0/ 0 0/ 0 0/ 0 0/ 0 418-014: PAGE B-3 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER; T - 6 2 9 S 1 3 ) TABLE 2 (PAGE 1): NECROPSY OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) ii 1.6 RATS EXAMINED a N 41b 36 MORTALITY N 00 APPEARED NORMAL N 40 36 LEFT INGUINAL AREA: MASS N i0 a. Refer to the individual clinical observations table (Table B15) for external observations confirmed at necropsy. b. Excludes values for dam 18933, which was discarded without further evaluation. 418-014: PAGE B-4 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 3 (PAGE 1) : BODY WEIGHTS - PRECOHABITATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) RATS TESTED N 42 BODY WEIGHT (G) DAY 1 M E A N + S .D 231.4 + 8.6 DAY 8 M E A N + S .D 246.5 4 10.0 DAY 15 M E A N + S .D 257.9 4 11.5 DAY 22 M E A N t S .D 266.3 4 14.2 DAY 29 M E A N + S .D 274.7 4 14.7 DAY 36 M E A N + S .D 285.7 4 16.5 DAY 43a MEAN+S.D 283.9 4 12.2 [ 17]b DAY = DAY OF STUDY [ ] = NUMBER OF VALUES AVERAGED a. Last value recorded before cohabitation. b. Excludes values for rats assigned to cohabitation on day 42 of study. II 1.6 36 231.2 4 9.2 246.8 4 10.5 257.9 14.1 265.4 4 11.9 270.5 4 12.7 278.3 4 14.7 274.4 4 15.9 ~ec30oo 418-014:PAGE B-5 9 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 418-014:PAGE B-6 <N CO DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) RATS TESTED N 42 BODY WEIGHT CHANGE (G) DAYS 1 - 8 M E A N + S .D . +15.1 7.0 DAYS 8 - 15 M E A N + S .D . +11.4 + 6.3 DAYS IS - 22 M E A N + S .D . +8.4 + 7.7 DAYS 22 - 29 M E A N + S .D . +8.3 + 4.8 DAYS 29 - 36 M E A N + S .D . +11.0 + 6.1 DAYS 36 - 43a DAYS 1 - 43a M E A N + S .D . M E A N + S .D . -2.6 + 7.7 f 17]b +53.0 + 11.4 l 17] b DAYS = DAYS OF STUDY [ ) = NUMBER OF VALUES AVERAGED a. Last value recorded before cohabitation. b. Excludes values for rats assigned to cohabitation on day 42 of study. II 1.6 36 +15.6 +11.0 + 7.5 +7.5 4.9 +5.1 + 5.9 +7.8 5-6 -3.9 + 6.2 +43.2 + 14.3 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 5 (PAGE 1): MATERNAL BODY WEIGHTS - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE (MG/KG/DAY) RATS TESTED N PREGNANT N MATERNAL BODY WEIGHT (G) DAY 0 M E A N + S .D . DAY 7 MEAN +S .D , DAY 14 M E A N + S .D . DAY 20 M E A N + S .D . DAY = DAY OF GESTATION I 0 (VEHICLE) 42 35 295.1 + 16.9 326.6 + 18.9 359.5 + 22.0 442.0 + 34.0 II 1.6 36 27 283.9 + 14.2 307.0 + 16.2 337.8 + 17.6 421.8 + 25.5 z o fre 418-014:PAGE B-7 C 01 * PROTOCOL 418-014: ORAL (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 DOSAGE (MG/KG/DAY) RATS TESTED N PREGNANT N MATERNAL BODY WEIGHT CHANGE (G) DAYS 0 - 7 M E A N + S .D . DAYS 7 - 14 M E A N + S .D . DAYS 14 - 20 M E A N + S .D . DAYS 0 - 20 MEAN+S. D. DAYS = DAYS OF GESTATION I 0 (VEHICLE) 42 35 +31.7 + 6.2 +32.7 + 7.4 +82.5 + 20.0 +146.9 + 24.8 II 1.6 36 27 +23.1 + 6.7 +30.7 + 8.7 +84.1 + 17.1 +137.9 + 19.9 9 Q B O OO 418-014:PAGE B-8 Sc u PROTOCOL 418-014: OR AL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 7 (PAGE 1): MATERNAL BODY WEIGHTS - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS SUBSET DOSAGE GROUP DOSAGE (MG/KG/DAY) A Ia 0 (VEHICLE) B Ib 0 (VEHICLE) C II c 1.6 RATS ASSIGNED TO CROSS-FOSTERING N 13 12 12 INCLUDED IN ANALYSES N lie 12 12 MATERNAL BODY WEIGHT (G) DAY 1 M E A N + S .D . 334.8 + 20.1 331.8 + 29.9 326.6 + 20.0 DAY 4 DAY 7 DAY 10 DAY 14 DAY 22 MEAN+S.D. M E A N + S .D . MEAN+S.D. MEAN+S.D. MEAN+S.D. 335.7 + 17.6 347.6 + 11.7 356.3 + 15.0 368.9 + 10.5 362.0 + 9.1 346.7 + 29.5 359.6 + 24.9 365.8 + 26.8 367.9 + 30.1 366.4 + 26.0 326.1 + 16.1 [ Ulf 334.2 + 18.2 [ 11) f 347.6 + 16.8 1 11) f 350.7 + 18.3 [ ll)f 354.7 + 19.1 DAY = DAY OF LACTATION [ ] = NUMBER OF VALUES AVERAGED a. Vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage gr ou p 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 grou p litters. e. Excludes values for dams which had litters that were not cr os s- f o s t e r e d . 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. D II d 1.6 13 lie 320.9 + 20.5 313.2 + 21.8 328.4 + 19.0 335.6 + 18.1 346.4 + 20.3 355.4 + 18.6 418-014:PAGE B-9 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 8 (PAGE 1): MATERNAL BODY WEIGHT CHANGES - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS SUBSET DOSAGE GROUP DOSAGE (MG/KG/DAY) A Ia 0 (VEHICLE) B Ib 0 (VEHICLE) C II C 1.6 RATS ASSIGNED TO CROSS-FOSTERING N 13 12 12 INCLUDED IN ANALYSES N lie 12 Ilf MATERNAL BODY WEIGHT CHANGE (G) DAYS 1 - 4 M E A N + S .D . +0.9 + 14.8 +14.9 + 12.4 +3.0 + 7.3 DAYS 4 - 7 M E A N + S .D . +11.9 + 10.7 +12.9 + 13.0 +8.1 + 6.7 DAYS 7 - 10 M E A N + S .D . +8.6 + 15.3 +6.2 + 8.6 +13.4 + 9.0 DAYS 10 - 14 M E A N + S .D . , +12.6 + 16.6 +2.1 + 13.7 +3.1 + 9.1 DAYS 14 - 22 M E A N + 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-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 gr ou p 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. D ii d 1.6 13 lie -7.7 + 14.8 +15.3 + 11.0 +7.2 + 7.2 +10.7 + 9.1 +9.0 + 15.8 +34.4 + 13.5 418-014PAGE B-10 to VA 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 - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) II 1.6 RATS TESTED N 42 36 FEED CONSUMPTION (G/DAY) DAYS 1 - 8 DAYS 8 - 15 DAYS 15 - 22 DAYS 22 - 29 DAYS 29 - 36 DAYS 36 - 43b DAYS 1 - 43b MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D, MEAN+S.D. .] 19.5 + 1.7 I 40] a 20.1 + 1.7 l 41] a 1 9 . 7 + 1.8 ( 41) a 19.6 + 2.2 l 40] a 19.6 + 2.1 l 40] a 18.3 + 2.4 1 15) a ,c 19.8 + 1.6 [ 15]a,c 19.7 + 2.0 [ 34] a 19.7 1.6 18.9 + 1.3 [ 34]a 18.7 + 1.3 ( 34] a 18.3 + 1.6 [ 34 ]a 17.2 + 1.5 l 33) a 18.7 + 1.3 [ 33] a DAYS = DAYS OF STUDY [ ] <= 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. 418-014:PAGE B-11 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 FEMALE RATS DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) II 1.6 RATS TESTED N 42 36 FEED CONSUMPTION (G/KG/DAY) DAYS 1 - 8 DAYS 8 - 15 DAYS 15 - 22 DAYS 22 - 29 DAYS 29 - 36 DAYS 36 - 43b DAYS 1 - 43b MEAN+S.D. M E A N + S .D . MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 81.6 + 6.1 ( 40) a 79.7 + 5.4 I 41) a 75.0 + 5.1 l 41) a 72.7 + 6.9 [ 401 a 70.0 + 5.8 [ 40) a 63.8 + 7.7 [ 15)a,c 74.2 + 4.9 [ 15}a,c 82.4 + 8.1 ( 34) a 78.0 + 5.2 72.1 + 4.6 [ 34 ]a 69.7 + 3.9 [ 34] a 66.8 + 4.6 [ 34]a 61.9 + 3.9 [ 33 ]a 71.7 + 3.8 ( 33] a DAYS = DAYS OF STUDY [ ] = NUMBER OF VALUES AVERAGED a. Excludes values Chat 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. oosooo 418-014-.PAGE B-12 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 FEMALE RATS DOSAGE GROUP DOSAGE (MG/KG/DAY) RATS TESTED N PREGNANT N MATERNAL FEED CONSUMPTION (G/DAY) DAYS 0 - 7 MEAN+S.D. DAYS 7 - 14 M E A N + S .D . DAYS 14 - 20 M E A N + S .D . DAYS 0 - 20 MEAN+S.D. DAYS = DAYS OF GESTATION l ] = NUMBER OF VALUES AVERAGED a. Excludes values that were associated with spillage. I 0 (VEHICLE) 42 35 2 4 . 2 + 2.7 ( 33 ]a 26.3 + 3.0 24.8 + 2.6 25.1 + 2.2 II 1.6 36 27 21.1 + 1.5 23.7 + 2.0 25.4 + 2.5 23.3 + 1.5 418-014:PAGE B-13 q cc ft 0! PROTOCOL 418-014: ORAL (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 GENERATION FEMALE RATS DOSAGE GROUP DOSAGE (MG/KG/DAY) RATS TESTED N PREGNANT N MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 0 - 7 MEAN+S.D. DAYS 7 - 14 M E A N + S .D . DAYS 14 - 20 M E A N + S .D . DAYS 0 - 20 MEAN+S.D. DAYS DAYS OF GESTATION ( ) = NUMBER OF VALUES AVERAGED a. Excludes values were associated with spillage. I 0 (VEHICLE) 42 35 76.9 + 5.8 [ 33] a 76.7 + 7.8 62.8 + 4.6 72.0 + 4.1 II 1.6 36 27 71.2 + 4.2 73.8 + 4.7 67.6 + 6.3 70.7 + 3.4 418-014:PAGE B-14 ri PROTOCOL 418-014: ORAL (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 FEMALE RATS SUBSET DOSAGE GROUP DOSAGE (MG/KG/DAY) RATS ASSIGNED TO CROSS-FOSTERING INCLUDED IN ANALYSES N N A Ia 0 (VEHICLE) 13 lie B Ib 0 (VEHICLE) 12 12 C II C 1.6 12 Ilf D II d 1.6 13 lie MATERNAL FEED CONSUMPTION (G/DAY) DAYS 1 - 4 M E A N + S .D . 19.1 + 4.0 24.9 + 3.6 18.8 + 3.3 21.3 + 3.9 DAYS 4 - 7 M E A N + S .D . 38.8 + 2.8 42.9 + 3.7 34.5 + 6.4 38.5 + 5.5 DAYS 7 - 10 DAYS 10 - 14h DAYS 1 - 14h M E A N + S .D . MEAN+S.D. MEAN+S.D. 45.0 + 3.3 [ 10) g 57.8 + 3.9 41.4 + 2.5 51.3 + 4.6 I nig 58.2 + 4.5 45.4 + 3.3 41.2 + 3.6 5 2 . 0 + 5.8 ( 10] g 38.0 + 4.4 [ 10] g 4 3 . 3 + 3.3 53.3 + 5.1 40.2 + 3.7 DAYS = DAYS OF LACTATION ( ] 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, were not tabulated on days 14 to 21 of lactation. maternal feed consumption values a e fro 418-014:PAGE B-15 c PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 14 (PAGE 1): MATERNAL RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS SUBSET DOSAGE GROUP DOSAGE (MG/KG/DAY) A 1a 0 (VEHICLE) B Ib 0 (VEHICLE) C II c 1.6 D II d 1.6 RATS ASSIGNED TO CROSS-FOSTERING N 13 12 12 13 INCLUDED IN ANALYSES N lie 12 Ilf lie MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 1 - 4 M E A N + S .D . 57.5 + 12.5 73.8 + 11.6 58.4 + 11.1 67.2 + 12.2 DAYS 4 - 7 M E A N + S .D . 113.2 + 9.3 122.0 + 15.1 104.4 + 18.6 119.1 + 12.3 DAYS 7 - 10 DAYS 10 - 14h DAYS 1 - 14h MEAN+S.D. MEAN+S.D. MEAN+S.D. 127.2 + 10.4 t 10] g 159.7 + 12.0 118.8 + 8.9 141.9 + 14.0 1 n)g 159.9 + 1 0 . 8 128.5 + 11.4 121.2 + 12.9 150.7 + 15.8 ( 10) g 113.8 + 12.6 l 10] g 129.8 + B .3 156.8 + 10.1 121.9 + 7.3 i DAYS = DAYS OF LACTATION ( ] = 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, were not tabulated on days 14 to 21 of lactation. maternal feed consumption values 418-014:PAGE B-16 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 15 (PAGE 1): NATURAL DELIVERY OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) II 1.6 RATS ASSIGNED TO CROSS-FOSTERING N 25 25 PREGNANT N(V) 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 OF DAMS DELIVERING DAYS 21.1 - 22.0 N (% ) DAYS 22.1 - 23.0 N (% ) 14( 60.9) 9( 39.1) 23(100.0) 0( 0.0) DURATION OF GESTATION IN WHOLE DAYS C M E A N + S .D . 22.4 + 0.5 22.0 + 0.0 NUMBER OF DAMS DELIVERING DAY 22 N (% ) DAY 23 N 1%) 14( 60.9) 9( 39.1) 23(100.0) 0( 0.0) GESTATION INDEX d % N/N 100.0 23/ 23 100.0 23/ 23 D A Y (S ) = D A Y (S ) OF GESTATION a. Excludes values ior 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 defined as 0 hour) 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 time (in days) the first pup was delivered. d. Number of rats with live offspring/number of pregnant rats. and 418-014: PAGE B-17 >-> PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 15 (PAGE 2): NATURAL DELIVERY OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE (MG/KG/DAY) I 0 (VEHICLE) II 1.6 RATS ASSIGNED TO CROSS-FOSTERING N 25 25 PREGNANT N (%) 25(100.0) 25(100.0) DELIVERED LITTERS N(%) 25(100.0) 25(100.0) INCLUDED IN ANALYSES N 23a 23a OBSERVED DURATION OF DELIVERY PER PUP PER LITTER (MINUTES) TOTAL DELIVERY PERIOD OBSERVED (MINUTES) b MEAN+S.D. TOTAL OR PARTIAL DELIVERY PERIOD OBSERVED (MINUTES) b M E A N + S .D . 11.6 + 4.8 ( 22) 11.6 + 4.8 9.1 + 3.1 9.1 + 3.1 IMPLANTATION SITES PER DELIVERED LITTER N M E A N + S .D . 407 17.7 + 1.8 368 16.0 + 1.9 DELIVERED LITTER SIZE MEAN+S.D. 16.4 + 1.8 15.0 + 1.8 DAMS WITH STILLBORN :PUPS N (%) 3( 13.0) M 4.3) PUPS FOUND DEAD (P R E C R O S S - F O S T E R I N G ) N/N(%) 1/373( 0.3) 1/344 ( 0.3) LIVE LITTER SIZE C M E A N + S .D . 16.2 + 1.8 14.9 + 1.8 FOSTER LITTER SIZE MEAN+S.D. 15.2 + 1.9 1 5 . 6 + 1.9 ( ] = 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 pups. c. Excludes values for dams with all pups dying prior to c r os s- fo st er in g. 418-014: PAGE B-18 * 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 DOSAGE GROUP DOSAGE (MG/KG/DAY) A Ia 0 (VEHICLE) B Ib 0 (VEHICLE) C II c 1.6 RATS ASSIGNED TO CROSS-FOSTERING N 13 12 12 PREGNANT N(%) 13(100.0) 12 (100.0) 12(100.0) DELIVERED LITTERS N (%) 13(100.0) 12(100.0) 12(100.0) INCLUDED IN ANALYSES N lie 12 12 LIVE LITTER SIZE e M E A N + S .D . 16.4 + 1.6 15.9 + 2.1 14.8 + 1.9 FOSTER LITTER SIZE M E A N + 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<%) 0( 0.0) 0( 0.0) 1< 8.3) DAMS WITH ALL PUPS DYING DAYS 5-21 POSTPARTUM N (% ) 0( 0.0) 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 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. D II d 1.6 13 13(100.0) 13(100.0) lie 15.1 + 1.7 16.4 + 1.6 0( 0.0) 0( 0.0) 418-014:PAGE B-19 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 - FI GENERATION LITTERS SUBSET DOSAGE GROUP DOSAGE (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 lie 12 12 PUPS CROSS-FOSTERED N M E A N + 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 VIABILITY INDEX f N / N (%) N / N (%) N / N (%) N / N (%) N / N (%) % N/N 0/1661 15/166( 1/109( 0/108( 0/1081 0.0) 9.0) 0.9) 0.0) 0.0) 91.0 151/166 0/191( 3/191( 0/120( 0/120( 0/1201 0.0) 1.6) 0.0) 0.0) 0.0) 98.4 188/191 0/177( 34/177( 0/107( 0/107( 0/107( 0.0) 19.2) 0.0) 0.0) 0.0) 80.8 143/177 LACTATION INDEX g t N/N 99.1 108/109 100.0 120/120 100.0 107/107 D A Y (S ) = D A Y (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 grou p dams cross-fostered with 1.6 mg/kg/day dosage grou p 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) postpartum/number of pups cross-fostered on day 1 postpartum. g. Number of live pup3 on day 21 postpartum/number of live pups on day 4 (postculling) postpartum. D II d 1.6 13 lie 181 16.4 + 1.6 0/181 ( 2/181( 0/110( 0/110( 0/110( 0.0) 1.1) 0.0) 0.0) 0.0) 98.9 179/181 100.0 110/110 8JKUTT 418-014:PAGE B-20 O VJ1 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 16 (PAGE 2): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - FI GENERATION LITTERS SUBSET DOSAGE GROUP DOSAGE (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 lie 12 12 SURVIVING PUPS/LITTER f DAY 1 M E A N + S .D . 15.1 + 1.7 15.9 + 2.1 14.8 + 1.9 DAY 4 PRECULLING M E A N + S .D . 13.7 + 2.3 15.7 + 1.8 11.9 + 4.6 DAY 4 POSTCULLING M E A N + S .D . 9.9 + 0.3 10.0 + 0.0 8.9 + 2.9 DAY 7 M E A N + S .D . 9.8 + 0.6 10.0 + 0.0 8.9 + 2.9 DAY 14 M E A N + 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 [ ] - 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. D II d 1.6 13 lie 16.4 + 1.6 16.3 + 1.6 10.0 + 0.0 10.0 + 0.0 10.0 0.0 10.0 + 0.0 418-014:PAGE B-21 1 t PROTOCOL 418-014: ORAL (GAVAGE) CROSS-POSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 16 (PAGE 3): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - FI GENERATION LITTERS SUBSET DOSAGE GROUP DOSAGE (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 lie 12 12 PERCENT MALE PUPS PER NUMBER OF PUPS SEXED DAY 1 MEAN+S.D. 46.9 + 9.7 48.4 + 14.4 48.8 + 16.6 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 46.7 + 11.0 48.7 + 6.4 49.3 + 7.4 49.3 + 7.4 49.3 + 7.4 49.1 + 14.3 52.5 + 8.7 52.5 + 8.7 52.5 + 8.7 52.5 + 8.7 49.4 + 21.5 ( lllf 46.0 + 16.9 1 lllf 46.0 + 16.9 l lllf 46.0 + 16.9 [ lllf 46.0 + 16.9 ( lllf DAY 1 M E A N + S .D . 15.1 + 1.7 15.9 + 2.1 1 4 . 8 + 1.9 DAY 4 PRECULLING DAY 4 POSTCULLING DAY 7 DAY 14 DAY 21 MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 13.7 + 2.3 9.9 + 0.3 9.8 + 0.6 9.8 + 0.6 9.8 + 0.6 15.7 + 1.8 10.0 + 0.0 10.0 + 0.0 10.0 + 0.0 10.0 + 0.0 13.0 + 2.9 [ lllf 9.7 + 0.6 [ lllf 9.7 + 0.6 [ lllf 9.7 + 0.6 ( lllf 9.7 + 0.6 I lllf DAY = DAY POSTPARTUM [ ] = 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 h a d litters that were not cross-fostered. f. Excludes values for dam 18974, which had no surviving pups on day 4 postpartum. D II d 1.6 13 lie 46.4 + 8.6 46.4 + 9.2 50.0 + 0.0 50.0 + 0.0 50.0 + 0.0 50.0 + 0.0 16.4 + 1.6 16.3 + 1.6 10.0 + 0.0 10.0 + 0.0 10.0 + 0.0 10.0 + 0.0 418-014:PAGE B-22 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 - FI GENERATION LITTERS SUBSET DOSAGE GROUP DOSAGE (MG/KG/DAY) A Ia 0 (VEHICLE) B ib 0 (VEHICLE) C II c 1.6 D II d 1.6 LITTERS ASSIGNED TO CROSS-FOSTERING N 13 12 12 13 INCLUDED IN ANALYSES N lie 12 12 lie PUP WEIGHT/LITTER (GRAMS) DAY lg MEAN+S.D. 5.6 + 0.3 6.2 + 0.5 5.7 + 0.2 6.3 + 0.6 DAY 4 PRECULLING DAY 4 POSTCULLING DAY 7 DAY 14 DAY 21 MEAN+S.D. MEAN+S.D. M E A N + S .D . M E A N + S .D . MEAN+S.D. 7.1 + 0.5 7.3 + 0.5 12.3 0.9 26.7 1.7 41.9 1.8 8.3 + 0.8 8.5 + 0.8 14.1 + 0.9 29.0 + 1.5 46.3 + 2.5 7.2 + 0.8 1 11] f 7.3 + 0.8 I 111 f 11.6 + 1.3 ! ll]f 2 4 . 6 + 2.4 [ lllf 38.9 + 3.8 [ H]f 7.9 + 0.6 8.1 + 0.6 12.8 + 0.8 26.2 + 2.2 41.9 + 2.1 DAY = DAY POSTPARTUM [ ] = 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 -f o s t er ed. 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 1pup weights forSubsets B and D reflect pups of vehicle treated dams. The effects of cross-fostering are not reflectedin thesepup body weights. 418-014: PAGE B-23 O oO 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 - SUMMARY - FI GENERATION PUPS SUBSET DOSAGE GROUP DOSAGE (MG/KG/DAY) LITTERS EXAMINED (N) A Ia 0 (VEHICLE) 13 B Ib 0 (VEHICLE) 12 C 11 c 1.6 12 TRANSIENT CLINICAL OBSERVATIONS: e TOTAL FREQUENCY (DAYS X PUPS)/LITTERS WITH OBSERVATIONS TIP OF TAIL, BLACK N/N 0/ 0 15/ 1 0/ 0 LABORED BREATHING N/N 0/ 0 0/ 0 0/ 0 DARK IN COLOR N/N 0/ 0 0/ 0 0/ 0 NOT NURSING N/N 2/ 2 0/ 0 2/ 1 PERSISTENT CLINICAL OBSERVATIONS: e TOTAL FREQUENCY (DAYS X PUPS) /LITTERS WITH OBSERVATIONS RIGHT HINDPAW, SECOND DIGIT MISSING N/N 0/ 0 0/ 0 21/ 1 TAIL, BENT N/N 0/ 0 11/ 1 0/ 0 N/N = TOTAL FREQUENCY (DAYS X PUPS)/LITTERS WITH OBSERVATIONS. a. Vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage grou p 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. D II d 1.6 13 18/ 1 1/ 1 1/ 1 0/ 0 0/ 0 0/ 0 418-014:PAGE B-24 f3 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 18 (PAGE 1): NECROPSY OBSERVATIONS - SUMMARY - FI GENERATION PUPS SUBSET MATERNAL DOSAGE GROUP MATERNAL DOSAGE (MG/KG/DAY) A Ia 0 (VEHICLE) B Ib 0 (VEHICLE) C II C 1.6 D II d 1.6 LITTERS EXAMINED N 13 12 12 13 PUPS FOUND DEAD e ,f (AFTER CROSS-FOSTERING) N 7 0 18 23 NO MILK IN STOMACH g N (%) 4( 57.1) 0( 0.0) 18(100.0) 20( 87.0) PUPS SACRIFICED AND NECROPSIED ON 4 OR 21 POSTPARTUM f LITTERS EXAMINED N 9 8 7 8 PUPS EVALUATED N 79 75 73 92 APPEARED NORMAL LITTER INCIDENCE PUP INCIDENCE N (%) N (%) 8( 88.9) 78( 98.7) 8(100.0) 75(100.0) 7(100.0) 73(100.0) 8(100.0) 92(100.0) KIDNEYS: RIGHT, PELVIS, SLIGHT DILATION LITTER INCIDENCE N(%) PUP INCIDENCE N (%) 1( 11.1) M 1.3) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) 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 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. Restricted to the pups in which complete necropsies were performed. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. f. Refer to the individual pup clinical observation table (Table B32) for external clinical observations confirmed at necropsy. g. Analysis restricted to pups found dead and necropsied. 418-014:PAGE B-25 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 19 (PAGE 1) : CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY RAT # DESCRIPTION 18901 18902 18903 18904 18905 18906 18907 18908 18909 18910 18911 18912 18913 18914 18915 18916 18917 18918 18919 18920 18921 18922 18923 18924 18925 18926 18927 DG ( 14 ) DS( 11- 17) DS ( 18 ) DS( 12- 20) DS( 21 ) DG ( 0- 17) DS( 11- 19) DS ( 20 ) DG ( 7- 14) DG ( 15 ) DS( 8- 22) DS ( 23 ) DS ( 10- 44) DG ( 0- 12) DG ( 13 ) DL ( 8- 18) DL( 19- 22) DL ( 21- 22) DS ( 29- 39) DG ( 21 ) DL ( 1- 14) NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMORHINORRHEA NO ADVERSE FINDINGS LOCALIZED ALOPECIA: HEAD ALOPECIA NO LONGER APPARENT NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS INCISORS: MISALIGNED INCISORS: REALIGNED INCISORS: MISALIGNED NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NECK: ABRASION (0.5 CM IN DIAMETER) ABRASION HEALED LOCALIZED ALOPECIA: BACK ALOPECIA NO LONGER APPARENT LOCALIZED ALOPECIA: HEAD ALOPECIA NO LONGER APPARENT LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS ALOPECIA NO LONGER APPARENT NO ADVERSE FINDINGS TAIL: AB RASION <1.0 CM IN LENGTH) TAIL: SCAB (0.5 CM X 0.3 CM) RIGHT FOREPAW: THIRD DIGIT SWOLLEN a NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: BACK PERINEAL HERNIA PERINEAL HERNIA NO ADVERSE FINDINGS DS DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION a. Observation confirmed at necropsy. 418-014:PAGE B-26 o o o a ii PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 19 (PAGE 2) : CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY RAT # DESCRIPTION 18928 18929 18930 18931 18932 18933 18934 18935 18936 18937 18938 18939 18940 18941 18942 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) DG( 2 ) DG( 6 ) DG( 13- 15) DG ( 17- 18) DL( 1- 22) DL( 7- 22) 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 CHROMORH INORRHEA DEHYDRATION CHROMODACRYORRHEA CHROMORHINORRHEA INCISORS : MISALIGNED a NO ADVERSE FINDINGS PROTRUDING XIPHOID a NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS DS * DAY OF STUDY DG - DAY OF PRESUMED GESTATION DL - DAY OF LACTATION a. Observation confirmed at necropsy. 418-014:PAGE B-27 000312 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 II 1.6 MG/KG/DAY RAT n DESCRIPTION 18943 18944 18945 18946 18947 18948 18949 18950 18951 18952 18953 18954 18955 18956 18957 18958 18959 18960 18961 18962 18963 18964 18965 18966 18967 18968 18969 18970 DS ( 36- 42) DS( 43 ) DS( 43- 45) DG ( 0- 4) DG( 5 ) DS ( 17 ) D M 1- 22) DL( 16- 22) DG ( 10- 17) DS( 8- 44) DG ( 0- 20) DL ( 1- 22) LEFT FORELIMB: ABRASION (0.4 CM IN DIAMETER) ABRASION HEALED LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS ALOPECIA NO LONGER APPARENT NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMORHINORRHEA NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS RIGHT FOREPAW: FOURTH DIGIT BROKEN a LOCALIZED ALOPECIA: LIMBS a NO ADVERSE FINDINGS LOCALIZED ALOPECIA: UNDERSIDE NO ADVERSE FINDINGS NO ADVERSE FINDINGS SCALY TAIL SCALY TAIL SCALY TAIL a NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION a. Observation confirmed at necropsy. 418-014:PAGE B-28 000313 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 19 (PAGE 4) : CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS DOSAGE GROUP II 1.6 MG/KG/DAY RAT # DESCRIPTION 18971 18972 18973 18974 18975 18976 18977 18978 DS< 27- 28) DG ( 5- 22) DG ( 15- 22) DL< 1- 2) DL( 3 ) DL( 1- 2) DL ( 3 ) DG ( 7- 25) DL ( i- 3) CHROMORHINORRHEA LOCALIZED ALOPECIA: NECK LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS ALOPECIA NO LONGER APPARENT LOCALIZED ALOPECIA: NECK ALOPECIA NO LONGER APPARENT NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS NO ADVERSE FINDINGS PROTRUDING XIPHOID a NO ADVERSE FINDINGS DS = DAY OF STUDY DG DAY OF PRESUMED GESTATION DL - DAY OF LACTATION a. Observation confirmed at necropsy. 418-014.PAGE B-29 000314 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 DOSAGE (MG/KG/DAY) RAT NUMBER DAY OF PREGNANCY DOSAGES NECROPSY STATUS ADMINISTERED OBSERVATIONS a I 0 (VEHICLE) 18901 - DL 22 p 87 ALL TISSUES APPEARED NORMAL. 18902 J DG 25 NP 69 ALL TISSUES APPEARED NORMAL. 18903 DL 22 P 86 ALL TISSUES APPEARED NORMAL. 18904 J; DL 22 P 85 ALL TISSUES APPEARED NORMAL. 18905 . 1 DS 72 NP 71 ALL TISSUES APPEARED NORMAL. 18906 ! DL 22 P 87 ALL TISSUES APPEARED NORMAL. 18907 I: DL 22 P 86 ALL TISSUES APPEARED NORMAL. 1B908 j, DL 22 P 87 ALL TISSUES APPEARED NORMAL. 18909 . V DL 22 P 85 ALL TISSUES APPEARED NORMAL. 18910 . DL 14 P 76 ALL TISSUES APPEARED NORMAL. 18911 ? i ; DL 14 P 80 ALL TISSUES APPEARED NORMAL. 18912 > J- DL 22 P 87 ALL TISSUES APPEARED NORMAL. 18913 4 DL 14 P 80 ALL TISSUES APPEARED NORMAL. 18914 , DG 25 NP 68 ALL TISSUES APPEARED NORMAL. 18915 J DL 14 P 76 ALL TISSUES APPEARED NORMAL. 18916 ! DG 25 P 70 ALL TISSUES APPEARED NORMAL. ) e UTERINE CONTENTS: ONE IMPLANTATIONS (ONE DEAD FETUS).b 18917 0 DL 22 P 87 ALL TISSUES APPEARED NORMAL. 18918 N. DL 22 P 87 ALL TISSUES APPEARED NORMAL. 18919 C N DL 22 P 87 ALL TISSUES APPEARED NORMAL. 18920 '1 : r DL 22 P 86 ALL TISSUES APPEARED NORMAL. 18922 N . DL 22 P 85 ALL TISSUES APPEARED NORMAL. 18923 X M DL 22 P 87 ALL TISSUES APPEARED NORMAL. 18924 ; DL 22 P 87 ALL TISSUES APPEARED NORMAL. 18925 <} B- DG 26 NP 69 ALL TISSUES APPEARED NORMAL. 18926 M. DL 14 P 81 LEFT INGUINAL AREA: MASS (2.5 CM X 2.0 CM X 1.5 CM).c ;-y 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 clinical ob se rv at io ns table (Table 19) for external observations confirmed at necropsy. b. The fetus had a large, black right: hindlimb and a black tail. Evidence of compression on the abdomen and hindlimbs. c. First observed at necropsy. 418-014:PAGE B-30 000315 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 DOSAGE (MG/KG/DAY) RAT NUMBER DAY OF PREGNANCY DOSAGES NECROPSY STATUS ADMINISTERED OBSERVATIONS a I 0 (VEHICLE) 18927 DL 14 P 81 ALL TISSUES APPEARED NORMAL. 18928 DS 72 NP 71 ALL TISSUES APPEARED NORMAL. 18929 DL 22 P 88 ALL TISSUES APPEARED 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 HERE NOT RECORDED. 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 necropsy. 418-014.PAGE B-: 000316 CO 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 - Fo GENERATION FEMALE RATS DOSAGE GROUP DOSAGE (MG/KG/DAY) RAT NUMBER DAY OF PREGNANCY DOSAGES NECROPSY STATUS ADMINISTERED OBSERVATIONS a 18943 DL 22 P 87 ALL TISSUES APPEARED NORMAL. 18944 DG 25 NP 71 ALL TISSUES APPEARED NORMAL. 18945 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. 18958 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 F E T U S E S .b 18978 DG 25 NP 69 ALL TISSUES APPEARED NORMAL. P = PREGNANT NP = NOT PREGNANT DS = DAY OF STUDY DG = DAY OP PRESUMED GESTATION DL = DAY OF LACTATION a. Refer Co the individual clinical observations table (Table 19) for external observations confirmed at necropsy. b. Appeared normal for their developmental ages and moderate degree of autolysis at time maternal death occurred. 418-014:PAGE B-32 000317 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PAGE 1) : BODY WEIGHTS - PRECOHABXTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT # DOSAGE GROUP I DAY 1 2 3 4 18901 18902 18903 18904 18905 18906 18907 18908 18909 18910 18911 18912 18913 18914 18915 18916 18917 18918 18919 18920 18921 18922 18923 18924 18925 18926 221. 238. 227. 228. 214 . 254. 226. 233. 226. 237. 234. 232. 214. 240. 230. 222. 219. 233. 228. 229. 228. 235. 224. 242. 233. 241. 222. 244. 233. 226 . 208. 249. 230 . 222. 228. 241. 223. 224. 220. 241. 228. 231. 225. 247. 227. 238. 221. 243. 230. 238. 223. 241. 229. 244 . 232. 228. 218. 255. 236 . 226. 222. 241. 237. 232. 225. 250. 234. 236. 230. 238. 232. 240. 231. 246. 229. 242. 236. 244. 229. 233. 240. 227. 224 . 260 . 240. 221. 231. 235. 238. 240. 221. 255. 238. 234. 233. 232. 234. 242. 236. 239. 234. 248. 239. 251. DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). 0 (VEHICLE) MG/KG/DAY 5 232. 239. 237. 232. 223. 268. 238. 234 . 237. 237. 239. 242. 226 . 255. 241. 235. 231. 245. 234. 240. 237. 243 . 232. 250. 239. 256. 6 238. 239. 247. 234 . 221. 256. 243. 233. 238. 236. 234. 235. 232. 253. 244 . 247. 241. 253. 234. 251. 230. 243. 235. 250. 237. 254. 7 250. 249. 257. 235. 236. 275. 250. 238. 234 . 247. 249. 248. 237. 268. 248. 255. 247. 256. 246. 259. 240. 252. 246. 259. 243. 257. 8 241. 237. 253. 238. 229. 271. 249. 234. 245. 238. 249. 250. 235. 272. 250. 248. 243 . 239. 243. 252. 243. 251. 242. 256. 250. 256. 9 239. 246. 246. 239. 226. 273 . 247. 237. 244 . 234 . 249. 249. 242. 270. 258. 246. 244. 248. 244 . 252. 238. 252. 238. 260. 243. 257. 10 246. 250. 252. 243 . 223. 269. 250. 232. 247. 245. 244. 243. 232. 267. 253 . 254 . 250. 251. 239. 258. 232. 254. 236. 255. 241. 260. 11 252. 250. 260. 241. 230. 275. 255. 236. 240. 241. 253. 253. 240. 273. 256. 256. 257. 253. 247. 260. 242. 260. 242. 267. 244. 261. 12 256 . 248. 262. 244. 233. 280. 256. 240. 248. 242. 262. 257. 247. 272. 260. 254. 256 . 240. 247. 262. 242. 261. 243. 267. 246. 265. 13 255. 252. 256. 248. 236. 283. 253. 236. 250. 242. 251. 255. 252. 278. 264 . 249. 258. 255. 248. 260. 239. 258. 245. 268. 248 . 268. 14 258. 253. 265. 248. 231. 280. 258. 237. 251. 237. 247. 251. 248. 277. 272 . 259. 257 . 260. 250. 265. 240. 262. 246 . 260 . 246 . 268 . IS 260. 256. 269. 245. 237. 284. 259. 242. 247. 248. 257. 257. 254. 280. 262. 265. 262. 262. 250. 267. 250. 272. 253. 273. 24 7. 265. 16 254 . 250. 267. 250. 236. 289. 258. 242. 251. 244. 258. 263. 257. 286. 270. 262. 262. 253 . 258. 270. 250. 268. 254. 275. 251. 270. 418-014:PAGE B-33 000318 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PAGE 2) : BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERA TI ON FEMALE RATS RAT tt DOSAGE GROUP I DAY 1 2 3 4 18927 18928 18929 18930 18931 18932 18933 18934 18935 18936 18937 18938 18939 18940 18941 18942 231. 245. 223. 234 . 234 . 229. 221. 234. 228. 234. 224 . 246. 230. 243. 228. 245. 224. 234. 218. 235. 228. 234 . 220. 241. 226. 239. 226. 234. 231. 238. 231. 250. 230 . 242 . 222. 236. 240 . 242 . 216. 240. 228. 234 . 225. 249. 225. 238. 237. 256. 235. 248. 224 . 244 . 246. 240. 218. 232. 229. 241. 230. 253. 235. 247. 238. 261. DAY - DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G) . 0 (VEHICLE) MG/KG/DAY 5678 230. 251. 225. 246. 247. 238. 219. 246. 232. 244 . 232 . 254. 243. 255. 237. 258. 225 . 246. 225. 250. 244 . 247. 211. 250. 236. 245. 234. 248. 241. 259. 232. 249. 240. 260. 237. 249. 264. 258. 225. 256. 246. 244 . 236. 260. 240. 256. 246. 256. 236. 256. 234. 253. 258. 243. 226. 247. 244 . 243 . 235. 259. 245. 258. 245. 257. 9 237. 263. 231. 258. 257. 243 . 227. 253. 244 . 254 . 238. 261. 244 . 265. 247. 254. 10 229. 249. 233. 262. 251. 249. 231. 260. 239. 252. 239. 253. 250. 265. 249. 247. 11 238. 252. 239. 256. 260. 255. 229. 265. 241. 250. 241. 262. 248. 259. 243. 255. 12 239. 260. 241. 262. 261. 259. 233. 258. 243. 256. 240. 273 . 252. 270. 248. 257. 13 239. 256. 240. 267. 262. 252. 233. 267. 245. 260. 240. 271. 257. 278. 253. 258. 14 232. 254. 236. 270. 256. 258. 233. 273 . 255. 261. 248. 269. 258. 278. 255. 254 . 15 242. 260. 244 . 264. 262. 267. 233. 272. 261. 255. 240. 272. 252. 269. 254. 261. 16 243 265 242 268 265 265 242 268 261 261 242 278 260 278 251 265 PROTOCOL 41B-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 I DAY 18901 16902 18903 18904 18905 18906 18907 18908 18909 18910 18911 18912 18913 18914 18915 18916 18917 18918 18919 18920 18921 18922 18923 18924 18925 18926 17 257. 258. 262. 254. 241. 293 . 259. 240 . 257. 249. 259. 262. 259. 286. 278. 259. 257. 249. 257. 262. 250. 263 . 250. 276. 257. 273. 18 259. 261. 268. 255. 235. 289. 264 . 243. 260. 246 . 253. 260 . 2S0 . 286. 277. 262 . 261. 257. 255. 270 . 246 . 272. 249. 276. 256. 274 . 19 20 265. 260. 271. 250. 242 . 300. 264 . 250. 256 . 246. 263 . 265. 259. 290. 270. 273. 266 . 262. 258. 269. 250. 276. 249. 284 . 248. 271. 260. 252. 267. 257. 239. 300. 267. 249. 251. 247. 266. 272. 263 . 294. 279. 268. 256. 262. 259. 265. 254 . 274. 256. 286. 254. 278. DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). 0 (VEHICLE) MG/KG/DAY 21 260. 257 . 266. 255. 243 . 301. 266 . 248. 260. 250. 261. 271. 258. 295. 280 . 266. 262. 248 . 259. 267. 254 . 270 . 253. 286 . 257. 280 . 22 266. 266. 277. 258. 238. 299. 271. 247. 262. 255. 262. 269. 259. 293 . 285. 269. 269. 245. 260 . 270. 248 . 274 . 257. 282. 260. 282. 23 267. 269. 278. 253 . 246. 307. 271. 248. 268. 252. 271. 266. 259. 292. 278. 271. 268. 256 . 263. 275. 252 . 278. 253. 289. 249. 276. 24 267. 261. 275. 253. 246. 306. 272. 252 . 255 . 250. 272 . 273 . 260. 296. 281. 268. 266. 255. 265. 271. 254. 276. 254 . 291. 252. 278 . 25 265. 265. 275. 257. 242. 308 . 271. 250. 268 . 256. 269. 278 . 263 . 299. 289. 260. 265. 258 . 268. 271. 254 . 278. 256 . 287. 255. 288 . 26 270. 270 . 282. 254. 238. 298. 275. 253. 267. 251. 271. 281. 256 . 296. 290. 266 . 268. 250. 264 . 275. 248. 280. 256. 283. 264 . 280. 27 274 . 271. 288. 261. 246. 309. 278. 257. 267. 251. 281. 279. 266. 302. 284. 269. 274. 262. 266. 277. 260. 282. 259. 296 . 266. 284 . 28 277. 268. 285. 261. 252. 316. 281. 264. 266 . 253. 273. 275. 271. 302. 290. 278. 275. 262. 271. 271. 262. 286. 260. 298. 261. 288. 29 275. 274 . 278. 262. 248 . 305. 279. 261. 264 . 258. 279. 284 . 266. 302 . 293 . 269. 276. 265. 268. 270. 262. 288. 259. 293 . 258. 291. 30 280. 280. 285. 260. 245. 308. 283 . 258. 266 . 263 . 274 . 287. 267. 301. 294 . 277. 276. 260. 265. 277. 258. 288. 263. 291. 266 . 287 . 31 282. 277. 287. 258. 253 . 317. 284. 261. 273 . 264 . 282. 285. 272. 307. 289. 276. 280. 256. 273. 276. 261. 288. 261. 299. 269. 288 . 32 278. 273. 288. 262. 258. 319. 284. 260. 272. 264. 282. 281. 278. 312. 297. 276. 282. 270. 271. 280. 265. 298. 266. 298. 269. 294. 418-014:PAGE B-35 000320 PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PAGE 4): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT It DOSAGE GROUP I DAY 17 18 19 20 18927 18928 18929 18930 18931 18932 18933 18934 18935 18936 18937 18938 18939 18940 18941 18942 241. 265. 245. 275. 264 . 262. 246. 280. 254 . 262 . 240. 279. 265. 286. 257. 263 . 241. 262. 246. 272. 261. 266. 247. 284 . 256. 267. 246. 272. 263. 286. 263. 262. 244. 268. 252. 269. 270. 267. 240. 281. 263. 260. 250. 277. 263. 279. 260. 266. 243 271 252 270 265 267 248 276 269 265 250 278 268 282 254 268 DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). 0 (VEHICLE) MG/KG/DAY 21 22 23 24 25 26 243. 269. 255. 277. 273 . 263. 249. 288. 267. 272. 247. 276. 270 . 293 . 255. 271. 242. 266. 253. 277. 268. 266. 254. 292. 263. 272. 246. 278. 269. 293. 262. 262. 246 276 261 275 266 270 252 290 267 268 249 278 268 279 266 269 243 . 273. 258. 281. 268. 270. 254. 283 . 264. 275. 249. 283. 271. 285. 269. 272. 243. 277. 260. 283 . 270. 265. 258. 294. 271. 276. 254. 283. 279. 301. 270 . 271. 240 268 256 283 270 270 254 296 262 274 247 276 277 294 262 262 27 242 . 280. 260. 280. 278. 278. 247. 295. 270. 271. 246. 284. 272. 284. 272. 275. 28 246 . 284 . 264 . 286. 280. 278. 256. 292. 275. 284. 249. 290. 278. 295. 275. 274 . 29 244 278 260 291 278 274 262 297 274 282 252 290 280 300 275 272 30 243 . 277. 257. 288. 278. 279. 260. 304. 272. 285. 256. 279. 278. 302. 274. 266. 31 246. 285. 263. 285. 283. 280. 259. 300. 275. 278. 250. 292. 276. 294 . 273. 276. 32 250 288 266 289 285 282 263 298 275 286 259 293 282 302 278 279 418-014:PAGE B-36 000321 PROTOCOL 418-014: ORAL GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T - 6 2 9 S .13) TABLE 21 (PAGE 5) : BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DA TA - Fo GENERATION FEMALE RATS RAT ft DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY DAY 33 34 35 36 37 38 39 40 18901 18902 18903 18904 1890S 18906 18907 18908 18909 18910 18911 18912 18913 18914 18915 18916 18917 18918 18919 18920 18921 18922 18923 18924 18925 18926 284 . 279. 282 . 267. 252 . 320. 287. 261. 274 . 260 . 283 . 278. 277. 310. 297. 278. 275. 275. 273. 273 . 268. 302. 265. 297. 260. 292. 284 . 283 . 292. 269. 2S2 . 317. 287. 261. 275. 264. 277. 286. 270. 307. 299. 277. 282. 273. 274 . 285. 262. 306. 267. 298. 273. 293. 293 . 285. 296. 265. 260. 325. 299. 266 . 274 . 273 . 292 . 290. 286. 316. 296. 288 . 286. 268 . 275. 285. 275. 302. 273 . 303. 279. 293 . 294 . 279 . 295. 268. 258. 328. 288. 266 . 281. 273 . 286. 287. 289. 318. 301 . 288. 289. 262. 280. 280. 273. 303. 268 . 307. 272 . 299. 291. 285. 291. 271. 260. 331. 286 . 262. 287. 269. 291. 290. 285 . 317. 304 . 286 . 283 . 276. 279. 278. 273 . 305. 269. 305. 267. 300. 294 . 286. 297. 272. 251. 320. 287. 257. 283 . 263 . 277. 286 . 271. 311. 302. 290. 280. 278. 274 . 281. 266. 305. 266. 297. 270. 299. 295. 282. 296. 270. 258. 324 . 292. 264 . 276. 269. 287. 293 . 280. 314. 299. 292. 287. 276 . 279. 285. 268 . 303. 273 . 307. 276. 300 . 293 . 278. 294. 271. 259. 332. 291. 264 . 281. 269. 291. 296. 286. 318. 305. 290. 284 . 266. 278. 281. 271. 300. 273 . 311. 274 . 299. 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. 41 288. 277. 287. 271. 255. 326 . 282. 262. 280. 268. 280. 294. 284. 316. 310. 281. 280. 260 . 276. 278. 269. 309. 268. 307. 266 . 302. 42 290. 282. 295. 272. 250. 319. 284 . 258. 280. 267. 280. 287. 277. 314. 308. 290. 286. 271. 272. 285. 263. 310. 271. 301. 276. 301. 43a b b 299. b b b b b b b b 288. 283 . b b 287. b 268. 272. 287. 262. b 268. b b 303. 418-014:PAGE B-37 000322 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 I 0 (VEHICLE) MG/KG/DAY DAY 33 34 35 36 37 38 39 40 18927 18928 18929 18930 18931 18932 18933 18934 18935 18936 18937 18938 18939 18940 18941 18942 250. 283. 266. 294. 283. 277. 268. 303. 278. 286. 255. 297. 287. 308. 276. 275. 247. 281. 266. 292 . 285. 281. 264. 310. 272 . 289. 260. 290. 285. 303. 283 . 273 . 254 . 292. 268. 291. 290. 291. 261. 312. 285. 285. 259. 293. 280. 298. 284 . 282. 258. 297. 273. 295. 294. 291. 262. 305. 286. 291. 254 . 304 . 294 . 302. 278. 282 . 259. 289. 274 . 299. 290. 286. 272. 315. 285. 293 . 264 . 303. 290. 314 . 284 . 287. 252. 285. 268. 298. 283. 288. 272. 315. 283. 262. 267. 298. 290. 307. 286. 276. 258. 292 . 274 . 298. 287. 290. 268 . 315. 286. 263. 263 . 296. 289. 298. 289 . 282. 259. 294 . 273. 299. 297. 290. 271. 308. 28S. 263. 262. 296. 291. 305. 285. 282. DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G) . a. Last value recorded before cohabitation. b. Rat assigned to cohabitation o n day 42 of study. 41 255. 289. 270. 304. 288. 286. 278. 318. 285. 263. 259. 300. 299. 310. 285. 281. 42 43a 251. 284. 267. 303 . 287. 292. 269. 323. 278. 267. 260. 298. 292. 313. 284. 278. b b b 290. 289. b b b 278 . 271. b 294. b 299. 288. b 418-014:PAGE B-38 000323 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 II DAY 1 2 3 4 18943 18944 18945 18946 18947 18948 18949 18950 18951 18952 18953 18954 18955 18956 18957 18958 18959 18960 18961 18962 18963 18964 18965 18966 18967 18968 218. 256. 234 . 236. 211. 250. 228. 234 . 227. 238. 235. 238. 225. 241. 226. 232. 222 . 246. 222. 225. 217. 237. 224 . 238. 226. 239 . 217. 266. 231. 238. 221. 246. 221. 230. 227. 244. 228. 233. 225. 239. 237. 234. 219. 245. 225. 231. 227. 240. 224. 239. 223 . 246. 227. 271. 229. 245. 228. 261. 216. 236. 225. 249. 237. 234. 220. 235. 242. 229. 226. 255. 226. 232. 230 . 240 . 224. 244 . 235. 247. 226. 270. 238. 248. 230. 257. 224 . 243. 229. 251. 242. 241. 228. 243. 240. 241. 228. 254 . 222. 241. 226. 234. 232. 242. 238. 243 . DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). 1.6 MG/KG/DAY 56 7 232. 263 238. 238. 228 261. 231. 245. 233. 244 . 241. 245. 233. 242. 234 . 247. 231. 262. 234. 236. 221. 244 . 230. 253. 236. 255. 224. 267. 244. 236. 239. 255. 229. 239. 237. 259. 237. 247. 236. 246. 249. 246. 230. 261. 236. 246. 233. 247. 233. 246. 233. 256. 234. 266. 236 . 241. 241. 264 . 226. 249. 229. 258. 242. 241. 227. 239. 249. 245. 230 . 269. 235. 252 . 241 . 244. 229. 248. 246. 259. 8 236. 263. 242. 244. 245. 268. 232. 251. 236. 261. 244 . 249. 233. 247. 249. 254 . 241. 272. 231. 253. 238. 237. 236. 254. 251. 254. 9 234 . 261. 246 . 242. 239. 261. 237. 257. 239. 255. 240. 250. 231. 251. 242. 258. 240. 272. 236. 252. 233. 243. 232. 254 . 249. 266. 10 229 . 268. 247. 238. 246. 250. 237. 248. 232. 265. 240. 249. 234. 250 . 252. 259. 244 . 272. 241. 259. 240. 240. 232. 259. 244. 264 . 11 238. 270 . 240. 246. 250. 253. 233. 257. 228. 268. 249. 247. 232. 252. 258. 251. 240. 275. 244 . 260. 242. 242. 236. 252. 257 . 268. 12 240. 268. 249. 247. 253. 270. 245. 262. 232. 269. 250. 245. 240. 255. 254. 260 . 237. 279. 237. 259. 241. 242. 237. 252. 259. 262. 13 236 . 262. 253. 248. 252. 262. 246. 264. 240. 262. 248. 251. 242. 261. 254. 264. 247. 277. 245. 258. 232. 248. 238. 256. 258. 270. 14 233. 272. 249. 245. 257. 263. 243. 253. 238. 279. 248. 256. 239. 261. 258. 268. 248. 277. 249. 263. 243. 253. 238. 262. 252. 273. 15 238. 272. 244. 252. 261. 269. 237. 262. 236. 281. 249. 254 . 240. 255. 265. 258 . 246 . 293 . 250. 272. 242. 251. 233 . 267. 261. 277. 16 240. 271. 2S0. 254. 263. 266. 248. 266 . 236. 280. 253. 253. 244 . 262. 262. 267. 243 . 291. 238. 269. 248. 246. 240. 261. 266. 272. 418-014:PAGE B-39 000324 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 ii DAY 1 2 3 4 18969 18970 18971 18972 18973 18974 18975 18976 18977 18978 229. 224 . 224. 234. 230. 236. 234. 232. 222. 234. 225. 234. 222. 239. 227 . 240. 231. 240. 229. 244 . 235. 235. 231. 245. 236. 250. 245. 247. 228. 243. 242. 238. 233. 247. 240. 250. 242. 249. 224. 240. DAY DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). 1.6 MG/KG/DAY 567 237. 233. 235. 243. 238. 246. 246. 246. 234. 252. 236 . 240. 234 . 250. 236. 254 . 242. 253 . 236. 252. 243 . 248. 243. 253 . 243. 262. 254. 246. 236. 254. 8 246. 245. 243. 250. 247. 263 . 256 . 229. 235. 251. 9 247. 243. 245. 254. 244 . 259. 257. 242. 238. 255. 10 244. 245. 239. 259. 242. 263. 250. 241. 242. 253. 11 252. 254. 245. 260 . 249. 271. 263. 241. 243. 254. 12 259. 254 . 252. 264. 252. 272. 266 . 243 . 240. 252. 13 256. 249. 250. 262. 249. 269. 266. 249. 245. 261. 14 253. 256. 246. 267. 249. 276. 263. 256 . 243 . 260 . 15 262. 261. 251. 273 . 256. 278 . 274 . 258. 247. 259. 16 261. 258. 253. 272. 259. 276. 280. 258. 247. 256. 418-014:PAGE B-40 000325 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 II DAY 17 18 19 20 18943 18944 18945 18946 18947 18948 18949 18950 18951 18952 18953 18954 18955 18956 18957 18958 18959 16960 18961 18962 18963 18964 18965 18966 18967 18968 240. 272. 253. 253. 258. 266 . 250. 266 . 243 . 270. 257. 253 . 243. 268. 254 . 274 . 243 . 285. 251 . 269. 241 . 255. 242. 259. 262. 279. 235. 272. 247. 256. 269. 260. 247. 261. 238. 283 . 254 . 256. 244 . 266 . 264 . 270. 249. 282 . 253 . 273. 252. 258. 239. 265. 261 . 281 . 241. 277. 247. 260. 268. 266. 247. 270 . 236. 287. 254 . 261 . 240. 263. 264 . 265. 253 . 296 . 252. 276. 253 . 258 . 237. 267. 264 . 280 . 244 . 274 . 251. 263 . 265. 267. 250. 270. 242 . 277. 258 . 259. 246 . 269. 260. 269. 253 . 288 . 249. 272. 249. 250. 246. 268 . 266 . 270. DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). 1.6 MG/KG/DAY 21 22 23 243. 276. 255. 260. 262. 268 . 259. 270. 243 . 268 . 262 . 258. 252 . 267. 255. 275. 250 . 290 . 252. 272. 241 . 261 . 242. 270 . 266 . 275 . 246. 280. 251. 264 . 269. 271. 252. 266. 244 . 286 . 259. 257. 253 . 267. 266. 274 . 250. 288. 258. 278. 254 . 260 . 242. 267 . 266. 280 . 249. 287. 246 . 261. 268. 273. 253. 268 . 241. 284 . 264 . 258. 251. 265. 263 . 268. 252. 293 . 252. 271. 256 . 258. 238 . 270. 274 . 276. 24 252. 284 . 250 . 269. 265. 278. 267. 268. 246. 283 . 267. 261. 265. 268. 262. 272. 254 . 294 . 244 . 273 . 254 . 246 . 244 . 272 . 275. 273 . 25 250. 278. 257 . 266 . 262. 279. 259. 271. 249. 276 . 269. 258. 258. 272. 258 . 275 . 256 . 296 . 252 . 269. 248. 257. 242 . 269. 273 . 285. 26 248. 287. 253. 264 . 264. 272. 258. 266. 246 . 283 . 269. 257. 258. 270 . 265. 276 . 250. 291. 257 . 282. 251. 263 . 244 . 271 . 272. 279. 27 252. 290. 250. 269. 265. 280. 253. 270. 246. 287. 278. 266 . 254. 271. 268. 268. 250. 299. 258. 287. 259. 263. 242. 274. 274 . 284. 28 252 . 290. 253 . 272 . 26S . 279. 262. 275. 251. 289. 278. 261. 263. 272 . 265. 278. 255. 303. 258. 287. 257. 261. 248. 275. 276. 278. 29 250. 290. 254 . 272. 262. 279. 264. 272. 254 . 282. 279. 261. 263. 280. 263. 280. 257. 298. 261. 283. 248. 263 . 245. 274 . 277. 286. 30 249. 288. 256. 269. 269. 274 . 258. 267. 256 . 292. 274. 259. 263. 276. 273. 279. 256. 297. 262. 290. 259. 262. 247. 276. 276. 289. 31 255 . 293 . 255. 269. 271. 279. 257. 270. 253 . 295. 280. 258. 262. 277. 272. 275. 254. 305. 260. 290. 259 . 260. 244. 274 . 282. 286 . 32 258 300 257 272 269 286 264 270 255 294 279 264 264 280 267 280 254 303 255 287 257 261 250 280 287 284 418-014-.PAGE B-41 000326 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PAGE 10): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT it DOSAGE GROUP II DAY 17 18 19 20 18969 18970 18971 18972 18973 18974 18975 18976 18977 18978 260. 256. 253. 271. 260. 266. 281. 258. 253. 266. 258 . 261. 250. 273. 254 . 274 . 273 . 260. 254 . 268. 267. 265. 255. 278. 257. 274 . 280. 262. 250. 264 . 264 . 264 . 259. 276 . 259. 276 . 286. 261. 248. 261 . DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G) . 1.6 MG/KG/DAY 21 22 23 265. 258 262 276. 259 272 285 260 258 274 266 . 268. 262. 279. 261. 280. 282. 271. 263 . 275. 271. 271. 260. 284 . 262. 284. 282. 268. 255. 266. 24 274 . 266. 261. 282. 262. 282. 291. 264. 252. 266. 25 271. 263. 264 . 281. 265. 279. 290. 265. 261. 273 . 26 271. 272. 263. 278. 262. 281. 287. 271. 263. 273. 27 28 29 30 31 274 . 274 . 268. 285. 266. 289. 290. 270. 262. 270 . 276. 272. 268. 289. 270. 283 . 296. 270. 255. 269 . 279. 269. 267. 286 . 267. 278. 291. 267. 265. 272. 270. 275. 264. 288. 261. 288. 288. 280. 264 . 275. 277. 276. 268. 290. 272. 286. 297. 279. 266. 274. 32 279. 279. 274. 295. 269. 285. 299. 275. 262. 274 . 418-014:PAGE B-42 000327 PROTOCOL 418-014: ORAL (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 n DOSAGE GROUP II 1.6 MG/KG/DAY DAY 18943 18944 18945 18946 18947 18948 18949 18950 18951 18952 18953 18954 18955 18956 18957 18958 18959 18960 18961 18962 18963 18964 18965 18966 18967 18968 33 257. 296. 257. 271. 265. 282. 271. 270. 255. 288. 283 . 268. 262. 286. 271. 285. 261. 309. 264. 288. 253. 267. 252. 280. 283. 292. 34 35 252 . 295. 255. 278. 274 . 284 . 263. 266. 256. 295. 280. 270. 263. 281. 275. 280. 261. 303 . 270. 288. 263. 271. 246. 280. 284 . 299. 263. 295. 257. 275. 276 . 288. 260. 265. 251. 301. 283. 268. 261. 283 . 276. 279. 264 . 314 . 270. 293. 267. 268. 247. 278. 288. 296. 36 262. 304. 259. 280. 274 . 289. 269. 273 . 260 . 299. 287. 264 . 265. 283 . 273 . 282 . 264 . 315. 265. 288. 266. 265. 250. 274 . 292. 286. 37 259. 309. 267. 281. 267. 286. 272. 276 . 258. 291. 288. 266. 266 . 287. 271. 284 . 266. 310. 270. 290. 263. 267. 248. 284. 294. 295. 38 250. 302. 264. 274 . 275. 280. 265. 272. 255. 295. 280. 265 . 258. 283 . 275. 284 . 272 . 307. 269. 289. 265. 266. 249. 282 . 287. 292 . 39 258. 300 . 256. 277. 276. 280 . 267. 266. 250. 299. 288. 266. 257. 279. 277. 279. 268. 313 . 264. 293. 265. 262. 246 . 282. 292 . 294 . DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. 40 257 . 305. 261. 283 . 273 . 290. 270. 270. 253. 294 . 286. 262 . 259. 282. 280 . 285. 268 . 313. 262. 289. 263. 258. 246. 279. 291. 289. 41 251. 303. 262. 282. 267. 281. 271. 271. 252. 284 . 287. 258. 259. 288. 272. 284 . 270. 308. 266. 287. 256. 260. 252. 282. 290. 293. 42 248. 298. 261. 280. 273 . 279. 268. 271. 252. 291. 283. 261. 259. 283. 278. 281. 274. 307. 270. 291. 258. 261. 248. 287. 285. 299. 43a 252. 292. 253. 281. 271. 288. 263. 269. 245. 288. 280. 254 . 253. 275. 275. 270. 271. 310. 269. 290. 259. 252. 240. 281. 288. 286. 418-014:PAGE B-43 000328 PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PAGE 12): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT # DOSAGE GROUP ii 1.6 MG/KG/DAY DAY 33 34 35 36 37 38 39 18969 18970 18971 18972 18973 18974 18975 18976 18977 18978 276. 272. 272. 291. 268. 282. 302. 275. 271. 278 . 275. 279. 270. 289. 268. 284 . 295. 281 . 272. 277. 286 . 285. 280. 295. 276 . 285. 308 . 279 . 276 . 275. 286 . 282. 280. 297 . 276. 282 . 309. 281. 267. 272. 286 . 277. 277. 296 277. 284 305 278 274 284 281. 280. 271. 293. 269. 286. 299 . 279. 276. 276. 288 . 281. 276. 297 . 274 . 292. 301. 283 . 275. 276. DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. 40 291. 281. 276. 298. 275. 284 . 302 . 278. 267. 265. 41 287. 274 . 274 . 295. 270. 278. 305. 274 . 272. 276. 42 285. 281. 275. 289. 269. 286. 301. 280. 276. 276 . 43a 284 . 282. 275. 291. 275. 288 . 301. 280. 273 . 275. 418-014:PAGE B-44 000329 418-014:PAGE B-45 000330 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 I 0 (VEHICLE) MG/KG/DAY PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 18901 P 18902 NP 18903 P 18904 P 18905 NP 18906 P 18907 P 18908 P 18909 P 18910 P 18911 P 18912 P 18913 P 18914 NP 18915 P 18916 P a 18917 P 18918 P 18919 P 18920 P 18921 NP 18922 P 18923 P 18924 P 18925 NP 293. 309. 311. 288. 300. 299. 296. 307. 313 . 276. 280 . 286. MATING NOT CONFIRMED 334. 348. 359. 301. 317. 320. 276. 287 . 282 . 290. 300 . 305. 270. 280. 287. 293. 308. 308. 302. 312. 316. 291. 299. 306. 307. 317. 332. 320. 326. 327. 286. 296. 296. 298. 308. 305. 279. 292. 293. 286. 300. 300. 294. 299. 303. 269. 266. 262. 310. 320. 318. 276. 282. 287. 323. 333. 341. 284 . 278. 284. 312. 303. 316. 292 . 366. 319. 287 . 304 . 286. 310. 319. 306. 329. 335. 296. 308. 299. 304. 307. 265. 325. 291. 346. 288. 321. 308. 325. 294 . 370 . 318 . 292. 308. 293 . 318. 321. 300. 335. 335. 310 . 314 . 301. 306. 310. 267. 329. 296. 355. 284 . 316. 314. 329. 299. 373. 330. 291. 310. 295. 316. 328. 300. 342. 340. 306. 317. 312. 315. 315. 263. 336. 298. 347. 275. 323. 316. 333. 301. 378. 329. 304. 317. 299. 322. 328. 302. 350. 346. 308. 323. 319. 320. 316. 259. 341. 299. 351. 289. 327. 318. 330. 307. 377. 327. 302. 320. 301. 327. 337. 314. 350. 345. 310. 326. 322. 320. 319. 262. 345. 302. 360. 285. 326 . 317. 335. 304. 384. 333. 307. 321. 304 . 330. 335. 309. 350. 354. 317. 328. 321. 329. 317. 263. 352. 301. 356. 287. 330. 321. 342. 308. 386. 336. 308. 325. 306. 335. 340. 313. 356. 358. 323. 330. 331. 326. 322 . 263. 357. 305. 360. 280. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION A L L 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. li 328 320 346 314 392 342 316 330 311 333 348 326 365 367 321 334 332 341 327 260 367 310 364 283 11 339. 318. 353. 316. 393 . 352. 324 . 340. 313. 337. 350. 329. 355. 377. 333. 345. 342. 350. 336. 266. 370. 318. 380. 291. 12 345. 327. 360. 321. 409. 357. 332. 345. 316. 345. 361. 334. 343. 382. 329. 348. 340. 353. 344. 266. 378. 321. 376. 291. 418-014:PAGE B-46 000331 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 22 (PAGE 2): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT # DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY PREGNANCY STATUS DAY 0 1 2 18926 P 18927 P 18928 NP 18929 P 18930 NP 18931 P 18932 P 18933 P 18934 P 18935 P 18936 P 18937 P 18938 P 18939 P 18940 P 18941 P 18942 P 317. 320. 327. 273 . 288. 293 . MATING NOT CONFIRMED 292. 302. 305. 312. 324. 329. 298. 300. 310. 290. 294. 304 . 288. 289. 293 . 315. 327. 336. 295 . 292. 297. 270. 272. 279. 265. 274 . 284 . 306. 317. 328. 302. 310. 313. 329. 333 . 340. 300. 308. 315. 294. 302. 305. 3 328. 298. 308. 332. 314. 307. 300 . 340. 306. 278. 289. 329 . 323. 346. 326. 312. 4 332. 303. 320. 334. 320. 312. 306 . 341. 319. 277. 294 . 328. 324. 357. 327. 315. 5 332. 304. 317. 334. 322. 315. 303. 351 . 321. 278. 300. 334. 326 . 369. 333. 321. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAM S (G) . 6 337. 304 . 323. 336. 330. 316. 304 . 349. 330. 277. 296. 335. 326. 363. 335. 321. 7 340. 315. 330. 338. 336. 316. 312. 354. 326. 291. 301. 341. 330. 360. 338. 330. 8 344. 313. 329. 337. 331. 321. 319. 361. 327. 305. 304 . 343. 334. 362. 343. 325. 9 345. 317. 335. 344 . 336. 328. 322. 368. 330. 313. 300. 351. 336. 370. 347. 328. 10 355. 320. 349. 343. 348. 330. 330. 368. 336. 312. 309. 358. 346. 376. 356. 337. 11 360. 327. 352. 344. 351. 340. 325. 375. 336. 318. 312. 361. 345. 383. 360. 341. 12 371. 327. 358. 352. 356. 344. 335. 373. 340. 324. 317. 366. 356. 390. 364. 346. 418-014:PAGE B-47 000332 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 22 (PAGE 3): MA TERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT # DOSAGE: GROUP I 0 (VEHICLE) MG/KG/DAY PREGI4ANCY STA'rus DAY 13 14 15 16 17 18 19 20 21 22 18901 P 18902 NP 18903 P 18904 P 18905 NP 18906 P 18907 P 18908 P 18909 P 18910 P 18911 P 18912 P 18913 P 18914 NP 18915 P 18916 P a 18917 P 18918 P 18919 P 18920 P 18921 NP 18922 P 18923 P 18924 P 18925 NP 344 . 353. 358. 325. 315. 310. 361. 367. 380. 322 . 326 . 335. MATING NOT CONFIRMED 422 . 414 . 426. 354 . 363. 376. 333. 339. 348. 349. 356. 362. 320. 319. 324 . 355. 354 . 365. 366 . 373. 374. 343. 343. 352. 340. 347. 341. 389. 397. 406. 331. 339. 340. 347. 358. 365. 359. 354 . 366. 366. 367. 366 . 348. 356. 364. 265. 259. 268. 374 . 383. 394. 32S. 333. 341. 393 . 388 . 394 . 280. 288. 294 . 374 . 309. 392. 356 . 440 . 380. 363 . 378. 333. 374 . 388. 368. 342. 426. 334 . 372. 369. 382. 372. 267. 404 . 348. 398. 289. 379. 309. 402. 371. 460 . 3 96. 377. 394 . 342. 392. 406. 383 . 346. 436. 334 . 386. 378. 3 94. 383 . 263 . 415. 359. 403 . 287. 400. 307. 426. 387. 483. 414 . 397. 395. 346 . 409. 427. 404 . 349. 452 . 340. 396 . 395. 413. 406. 258. 431. 376. 421 . 286 . 417. 300. 452 . 406 . 499. 436. 413. 414 . 355. 424 . 444 . 409. 346. 478. 347. 412. 413 . 434 . 428. 263. 442. 394 . 442. 287. 436. 304. 470. 427. 523. 456. 44 3 . 427. 357. 455. 467. 434 . 340. 491. 341. 428. 425. 454 . 434. 270. 465. 405. 468 . 298. 455. 313. 486. 450. 526. 478. 459. 445. 369. 464 . 480. 443 . 350. 525. 341 . 444 . 442 . 463 . 451. 271. 477. 412. 476. 298. 314. 304 . 429 . 329. 457. 347. 336. 438. 431. 268. 294 . . PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) ' = DAY OF PRESUMED GESTATION , WEIGHTS WERE RECORDED IN GRAMS (G). Dam 18916 did not deliver a litter; only one dead fetus was present in utero on day 25 of q e a t a H o n . 23 306. 351. 322. 277. 292. 24 303 . 25 --- 315. 350. 324. 350 377. 280. 286. 288. 290. -- 418-014:PAGE B-48 000333 PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 22 (PAGE 4) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT tt DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY PREGNANCY STATUS DAY 13 14 15 16 17 18 18926 P 18927 P 18928 NP 18929 P 18930 NP 18931 P 18932 P 18933 P 18934 P 18935 P 18936 P 18937 P 18938 P 18939 P 18940 P 18941 P 18942 P 373. 370. 375. 339. 339. 336. MATING NOT CONFIRMED 370. 375. 373. 338. 329. 329. 357. 357. 360. 348. 354. 366 . 337. 340. 350. 384. 385. 401. 358. 359. 370. 319. 330. 343. 323. 324 . 328. 378. 382. 385. 358. 360. 366 . 3 90. 392. 393. 372 . 370. 377 . 353. 364 . 365. 389. 353. 382. 335. 371. 368. 370 . 410. 382. 352. 343 . 401. 376. 408. 393. 373. 406. 364. 405. 335. 389. 383 . 384 . 413. 392. 357. 359. 417 . 389. 425. 400. 386. 420. 385. 423 . 328. 405. 398. 396. 434 . 417. 370. 374. 438. 395. 435. 424. 401. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). 19 441. 402. 437. 334. 405. 421 . 421. 455. 425. 389. 394. 446. 409. 456. 436. 420. 20 450. 429. 470 . 335. 428. 434 . 442. 475. 450. 398. 418. 472. 428 . 468. 466. 437. 21 465. 443 . 478. 327. 434. 457. 468. 480. 457 . 413. 427. 474 . 462. 469. 470. 445. 22 477 . 322. 449. 433. 310. 23 336. 24 338. 25 330. 418-014.PAGE B-49 000334 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 22 (PAGE S) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT # DOSAGE GROUP II PREGNANCY STATUS DAY 0 1 2 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 18967 P 261. 272 . 270 . 307. 319. 322. MATING NOT CONFIRMED 291. 295. 302. 282. 286. 293. 292. 300. 304 . 278. 279. 280. 287. 290. 294 . 264 . 264 . 272. 296. 307. 307. 295. 305. 306. 266. 268. 266. 261. 266. 267 . 288. 296. 298 . 289. 288. 293. 289. 304. 306. 270. 273 . 280. 316. 331. 327. 273. 279. 281. 290. 283. 287. 266. 268. 277. 259. 267. 272. 264 . 268. 269. MATING NOT CONFIRMED 285. 282 . 299. 1-6 MG/KG/DAY 3 272. 326. 303 . 292. 302. 284 . 296 . 273 . 310. 309. 261. 264 . 293 . 295. 308. 283. 336. 291. 288. 281. 272. 271. 303. 4 274 . 326. 304 . 297. 313 . 284 . 299 . 273. 316. 304 . 258 . 269. 295 . 296. 311. 280. 338. 298. 282. 288. 272. 272. 307. 5 276. 322. 298. 300. 314. 280. 296. 275. 319. 308. 263. 272. 290. 297. 310. 287. 334. 294. 281. 292. 276 . 278. 302. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). 6 276. 334. 311. 304 . 317. 279. 303. 280. 323. 316. 265. 268. 296. 301. 310. 284. 337. 302. 282. 293. 279. 275. 301. 7 276 . 340. 323. 309. 327. 287. 300. 284. 320. 316. 263. 264. 299. 301. 313. 290. 341. 304. 284 . 296. 284. 280. 306 . 8 282. 329. 314. 313. 325. 283 . 309. 285. 330. 320. 261. 267. 303. 308. 316. 293. 338. 302. 280. 302. 291. 281. 309. 9 282. 333. 315. 318. 329. 277. 307. 289. 329. 320. 265. 271. 305. 313. 322. 296. 346. 304. 275. 305. 287. 285. 310. 10 295. 340. 320. 321. 342. 278. 317. 294 . 331. 329. 263. 271. 309. 317. 330. 301. 352. 314. 281. 313 . 294 . 293 . 323 . 11 297. 348. 325. 319. 349. 282. 320. 299. 342. 337. 266. 267. 313. 324 . 336. 306 . 356. 317. 282. 320. 295. 302. 327 . 12 303. 349. 333. 330 . 354. 279. 320. 302. 341. 339. 260. 273 . 318. 327. 344 . 310. 360. 322. 282. 328. 302. 306. 332. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 22 (PAGE 6): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT # DOSAGE GROUP II 1.6 MG/KG/DAY PREGNANCY STATUS DAY 0 i 2 3 4 5 18968 P 18969 P 18970 P 18971 P 18972 P 18973 P 18974 P 18975 NP 18976 P 18977 P 18978 NP 302. 294. 286. 282 . 298. 268. 297. 313. 292. 280. 283. 302. 306. 292. 288 . 309. 279. 305. 315. 296. 285. 285. 307. 306. 295. 288. 309. 276 . 305. 318. 295. 283 . 284. 306. 305. 297. 290. 314. 282. 306. 324. 294. 289. 288. 311. 310. 294 . 291. 315. 280. 312. 315. 302 . 294 . 290. 310. 311. 298. 303. 315. 288. 319. 316. 306. 299. 292. P PREGNANT NP NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). 6 318. 310. 301. 300. 321. 288. 319. 325. 306. 308. 296. 7 328. 322. 300. 309. 324. 293. 319. 330. 311. 315. 298. 8 327. 316. 310. 303. 335. 291. 330. 326. 318. 316. 302 . 9 331. 322. 309. 304. 328. 294 . 328. 326. 320. 315. 301. 10 324. 331. 316. 315. 337. 303. 334. 333. 328 . 322. 294. 11 335. 340. 320. 312. 344. 308. 344 . 341. 330 . 329. 293. 12 341. 334. 323. 326. 349. 314. 348. 337. 335. 337. 283 . 418-014:PAGE B-50 o o o CJ CO C/l 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 II 1.6 MG/KG/DAY PREGNANCY STATUS DAY 13 14 15 16 17 18 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 18967 P 302. 307. 324 . 353. 336. 334 . MATING NOT CONFIRMED 335. 334 . 343 . 327. 342. 346. 362. 359. 359. 279. 281. 275. 318. 335. 341. 306. 309. 317. 340. 354. 359. 348. 351. 349. 255. 265. 264. 273. 272. 270. 320. 324. 330. 330. 338. 345. 346. 349. 356. 320. 320. 332. 367. 362. 371. 333. 328. 340. 279. 280. 284. 329. 339. 345. 304 . 309. 319. 310. 316. 332. MATING NOT CONFIRMED 345. 349. 352. 339. 327. 365. 356. 385. 280. 355. 330. 364. 366. 265. 272. 337. 350. 367. 341 . 388. 352. 284 . 355. 323. 345. 368. 354 . 328. 375. 374 . 402. 272 . 375. 346. 386. 370. 259. 268. 353. 369. 385. 351. 406 . 357. 285. 365. 338. 371. 388. 374 . 326. 394 . 394. 424. 281. 394 . 359. 402. 401. 255. 266. 366 . 389. 403 . 361. 425. 382. 288. 380. 361. 384. 406. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). 19 394. 321. 418. 405. 443 . 277. 410. 375. 420. 413. 263. 296. 380. 401. 421. 376. 438. 401. 288. 378. 374. 407. 425. 20 417. 325. 444 . 412. 458. 280. 419. 393. 430. 444 . 266. 269. 395. 413. 426. 394 . 449. 417. 290. 391. 391. 411. 445. 21 424 . 327. 450. 431. 470. 271. 442. 406. 446. 442. 254. 273. 412 . 420. 443. 406 . 461. 436. 288. 406. 399. 457. 22 325. 272 . 266. 274. 416 . 293 . 23 322 . 276. 255. 269. 294 . 24 329. 275. 254. 271. 297. 25 328. 278. 264. 277. 293 . 418-014.PAGE B- 000336 cn 418-014:PAGE B-52 000337 PROTOCOL 418-014: 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 n DOSAGE GROUP II 1.6 MG/KG/DAY PREGNANCY STATUS DAY 13 14 15 16 17 18 18968 P 18969 P 18970 P 18971 P 18972 P 18973 P 18974 P 18975 NP 18976 P 18977 P 18978 NP 342 . 345. 327. 322 . 365. 321 . 354 . 326 . 338. 341 . 278. 351. 350. 333. 320. 365. 321. 367. 315. 343. 345. 281. 366. 364. 334. 324 . 365. 329. 374 . 316. 352. 358. 291. 373 . 372. 346. 319. 383. 341. 378. 324 . 360 . 370 . 284 . 387. 386. 362. 332. 398. 355. 396. 317. 372. 380. 274. 407. 415. 374. 340. 422. 387. 410. 313 . 389. 3 94. 287. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G) . 19 426. 434 . 391. 345. 437. 398. 433. 318. 404 . 414 . 291 . 20 447. 459. 406. 354. 460. 422 . 440. 329. 423. 430. 288. 21 452. 472 . 406. 355. 468 . 420. 469. 309. 456. 461. 283 . 22 356. 309. 295. 23 318. 297. 294 . 24 320. 291. 25 311. 290. 000338 PROTOCOL 418-014: ORAL (GAVAGE) 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 ft DOSAGE GROUP I a 0 (VEHICLE) MG/KG/DAY SUBSET A 18903 18907 18908 18909 18918b 18920 18924 18929 18934 18936b 18937 18941 18942 DAY 1 353. 337 . 318. 319. 300 . 325. 351 . 330. 362. 315. 294. 354 . 340. 2 342 . 334 . 311. 319. 309. 325. 343 . 337. 353. 302. 303. 356. 332. 3 337. 329. 324 . 317. 321. 320. 318. 344 . 358. 310. 304 . 348. 337. 4 333 . 328. 321. 321. 341. 319. 356. 348. 365. 314 . 324 . 358. 320. 5 338. 345. 318. 337. 345. 338. 361. 353. 365. 303. 328. 358. 345. 6 341. 338. 319 . 336. 343. 339. 360. 354 . 373. 306. 340. 360. 347. 7 336. 346. 328. 347. 353. 345. 369. 350. 363. 326 . 344 . 356. 340. 8 345. 341. 336. 362. 360. 342. 368. 354. 369. 335. 358. 357. 3 54. 9 346. 353. 339. 345. 350. 307.C 370 . 365. 367. 332. 361. 365 . 347. DAY 14 15 16 17 18 19 20 21 22 18903 18907 18908 18909 18918 18920 18924 18929 18934 18936 18937 18941 18942 374. 355. 364 . 353 . 377. 360. 370. 383. 370 . 330. 381. 382. 366. 386 . 346. 370. 373 . 363 . 371. 388. 388 . 380 . 350. 371. 383. 378. 378 . 353. 368. 371 . 374 . 366 . 404 . 383 . 381 . 348. 377. 375 . 383 . 388 . 370. 368. 384 . 382. 363 . 394 . 390 . 390. 352. 368 . 376 . 372 . 383. 358. 367. 373. 386 . 367. 403 . 394 . 381 . 313 . 377. 403 . 361. 399. 351. 379 . 376. 388. 369. 396 . 380. 381. 335. 369. 391. 371. 390. 355. 374 . 379. 397. 374 . 398. 379. 372 . 326 . 392. 374. 372. 377. 349. 379. 386. 365. 370. 389. 380. 361. 349. 384 . 371. 364 . 364. 358. 372. 353 . 360 . 362. 365. 354 . 375. 353. 346 . 373. 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 cross-fostered; values excluded from group averages. c. Water access was interrupted; value excluded from group averages. 10 365. 366. 342. 359. 356. 345. 379. 371 . 335. 343. 342. 371. 344 . 11 363 . 357. 341. 350. 347. 360. 387. 356. 369. 333. 342. 383. 350. 12 368. 325. 337. 355. 351. 360. 373. 373. 378. 334. 362. 374. 368. 13 358. 356. 365. 345. 370. 350 . 376. 375. 379. 334 . 371. 374. 361. es-a 3 9 V d :n 0 -8 l.fr 418-014:PAGE B-54 000339 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T - 6 2 9 5 .13) TABLE 23 (PAGE 2) : MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RATS ASSIGNED TO CROSS-FOSTERING RAT it DOSAGE GROUP I a 0 (VEHICLE) MG/KG/DAY 18901 18904 18906 18912 18917 18919 18922 18923 18931 18932 18935 18938 18901 18904 18906 18912 18917 18919 18922 18923 18931 18932 18935 18938 DAY 1 313. 311. 399. 347. 325. 338 . 366 . 280. 324 . 320. 320. 338. DAY 14 384. 315. 436. 379. 370. 372. 377. 331. 366. 353. 351. 381. 2 338. 314 . 394 . 355. 331. 324 . 362 . 292. 322. 322. 313. 339. 15 376 . 333. 438. 382. 366. 380. 387. 339. 367. 352. 343 . 388. 3 343. 313 . 407. 355. 338. 344 . 370. 300. 331. 337. 320 . 346. 16 375. 343. 456 . 372. 377. 374. 389. 340. 363 . 367. 346. 397. 4 351. 314 . 420. 361. 343. 355. 363 . 307. 340 . 344 . 316. 346. 17 372 . 350. 457. 378. 364. 374 . 385. 343. 362. 362. 352. 390. 5 366. 330. 426. 363. 349. 354. 367. 314 . 345. 342. 339. 351. 18 388. 352. 441. 380. 374 . 380. 389. 339. 372 . 375. 348. 395. 6 368. 345. 420. 362. 352. 357. 341. 317. 358. 334. 334. 366. 19 385. 352. 443. 389. 372. 385. 390. 346. 373. 363. 352. 397. 7 370. 353 . 417. 368 . 357. 357. 372. 330. 356. 341. 319. 375. 20 395. 351. 446. 398. 383. 366 . 392. 334 . 375. 368 . 333. 383. 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. SUBSET B 89 355. 362. 391. 374 . 362. 349. 377. 323. 361. 348. 327. 370. 359. 353. 405. 376. 362. 369. 371. 330. 362. 355. 328. 382 . 21 22 380. 358. 437. 385. 377. 380. 401. 334. 370. 353. 340. 365. 358. 356. 406 . 345. 376. 375. 420. 324. 358. 368. 349. 362. 10 375. 341. 430. 383. 370. 359. 374. 344. 354. 359. 323. 378. 11 374 . 316.b 412. 363 . 368. 352. 372. 346. 355. 354. 326. 384 . 12 366. 337. 426. 366. 362. 353. 378. 336. 357. 349. 325. 375. 13 371. 337. 440. 367. 369. 375. 386. 328. 363. 347. 343. 392. 418-014PAGE B-55 000340 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 H DOSAGE GROUP II a 1.6 MG/KG/DAY SUBSET C DAY 1 2 3 4 5 6 7 8 9 18943 18947 18948 18957 18960 18964 18967 18968 18969 18970 18973 18974 291. 324. 345. 318. 345. 313. 331. 331. 336. 324 . 296. 360. 297. 318. 344 . 316. 349. 302. 327. 327. 334. 311. 292. 353 . 307. 325. 343. 313. 341. 313. 335. 331. 334. 316. 294 . 338. 310. 322. 341. 312. 352. 312. 342. 332. 338. 324. 302. 336. 325. 320. 324. 322. 326. 334. 338. 326. 344. 348 . 346. 350. 353. 364. 357. 317. 326. 325. 323. 332. 351. 354 . 361. 364 . 380. 319. 304 . 317. 325. 334 . 336. 331. 338. 361. 345. 324 . 340. 348. 344. 337. 342. 339. 352. 347. 358. 326. 329. 332. 345. 345. 300 . 290 . 300. 304. 312. NO SURVIVING PUPS ON DAY 4 OF LACTATION DAY 14 15 16 17 18 19 20 21 22 18943 18947 18948 18957 18960 18964 18967 18968 18969 18970 18973 18974 344 . 334. 362. 358. 353. 359. 359. 368. 376. 372 . 374 . 378 . 385. 372. 380. 349. 361. 357. 360. 366. 381. 380. 380 . 395. 399. 329. 343. 348. 348. 353. 346. 374 . 374 . 379. 383. 350. 364. 363. 377. 381. 360. 369. 365. 371. 372. 350. 358. 364 . 374. 370 . 316. 314 . 315. 324. 327. NO SURVIVING PUPS ON DAY 4 OF LACTATION 356. 368. 378. 366 . 397. 351. 383. 373. 371. 371. 332. 289. b 367. 326. 353. 407. 363. 383. 388. 361. 376. 332. 322. 371. 292.b 352. 396. 361. 357. 385. 368. 344.b 317. 339. 339. 353. 350. 390. 329. 366. 374 . 376. 343 . 343 . DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. 1.6 mg/kg/day dosage gcoup dams cross-fostered with 1.6 mg/kg/day dosage gr oup litters. b. Water access was interrupted; value excluded from group averages. 10 331. 344. 363. 345. 376. 330. 360. 347. 351. 359. 318. 11 316. 347. 363. 337. 380. 327. 348. 358. 354. 348. 317. 12 324. 350. 357. 338. 376. 329. 367. 353. 357. 355. 306. 13 346. 346. 366. 341. 330.b 321. 368. 359. 366. 340. 314 . 418-014:PAGE B-56 000341 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 0 DOSAGE GROUP II a 1.6 MG/KG/DAY SUBSET D DAY 1 2 3 4 5 6 7 8 9 18946b 18950 18951 18952 18953 18958 18959 18961 18963 18965 18972 18976 18977b 325. 332. 299. 328. 325. 333 . 307. 327. 298. 286 . 349. 346. 354. 321. 326 . 298. 331. 320. 327. 295. 311. 293 . 295. 344 . 323. 338. 320. 319. 300. 333 . 323. 327. 291. 315. 298. 305. 351. 325. 334. 322. 329. 334. 342. 320. 317. 328. 344 . 332. 342. 300. 307. 305. 305. 309. 335. 350. 355. 350. 358. 318. 316. 315. 321. 332. 324 . 316. 316. 321. 326. 290. 291. 306 . 315. 323. 286. 306. 320. 320. 327 . 296. 309. 308 . 311. 324 . 295. 308. 321. 323 . 328 . 356. 363. 370 . 368. 366. 328. 342. 348. 347. 349 . NO SURVIVING PUPS ON DAY 3 OF LACTATION 343. 351. 311. 355. 327. 329. 318. 324. 324 . 330. 368. 358. DAY 14 15 16 17 18 19 20 21 22 18946b 18950 18951 18952 18953 18958 18959 18961 18963 18965 18972 18976 18977b 350. 354. 357. 343. 351 , 353 . 353. 365. 354. 362 , 334. 335. 338. 346. 346 . 360. 364 . 351. 365. 348. 340. 347. 356. 368. 358 . 344 . 343. 352. 347. 368 . 310. 319. 319. 334. 336. 334 . 342. 341. 354 . 354. 346. 352. 352. 352. 351. 334. 345. 347. 352 . 344 . 385. 376. 386. 389. 393. 370. 362. 361. 387. 368. NO SURVIVING PUPS ON DAY 3 OF LACTATION 352. 358. 345. 338. 358. 367 . 337. 357. 354 . 348 . 400. 384. 354. 365. 320. 352, 363. 359. 340. 354 362. 348 328 373 365. 361. 330. 362. 360. 361. 335. 353. 362. 336. 310. 374 . 338. 361. 347. 344 . 360. 371 . 346. 333. 340. 337. 380 . 390. 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 grou p averages. 10 346. 335. 311. 360. 339. 331. 309. 330. 328. 332. 368. 349. 11 352. 337. 309. 360. 348. 329318. 341. 340. 322. 370. 339. 12 341. 349. 317. 365. 331. 335. 315. 343 . 325. 344 . 372 . 350. 13 348. 354. 325. 357. 340. 338. 306. 335. 342. 291. 372. 355. 418-014:PAGE B-57 000342 PROTOCOL 418-014: ORAL (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 - Fo GENERATION FEMALE RATS RATS ASSIGNED TO PHARMACOKINETIC EVALUATION RAT # DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY DAY 1 2 3 4 5 6 18910 18911 18913 18915 18926 18927 18939 18940 311. 348. 320. 380. 371. 312. 341. 343. 297. 347. 323. 365. 364. 309. 328 . 357. 296. 348. 328. 370 . 366 . 322. 339. 351. 303 352 324 370 375 322 342 370 313. 369. 348. 374. 376. 331. 350 376. 317. 380. 354. 379. 376. 330. 352. 353. DAY 14 18910 18911 18913 18915 18926 18927 18939 18940 332. 363 . 368. 370 . 389. 345. 361. 403 . DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). 7 330. 371. 336. 395. 374 . 333. 359. 358. 8 329. 370. 345. 406. 386. 337. 357. 365. 9 330. 368. 339. 393. 397. 344 . 352 . 372. 10 338. 355. 342. 396. 393. 348. 357. 373. 11 329. 362. 350. 408. 401. 330. 365. 368. 12 338. 365. 350. 399. 401. 344 . 367. 391. 13 335. 366. 356. 394 . 401. 341. 367. 398. 418-014:PAGE B-58 000343 PROTOCOL 418-014: ORAL (GAVAGE) CROSS -POSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 23 (PAGE 6) : MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RATS ASSIGNED TO PHARMACOKINETIC EVALUATION RAT It DOSAGE GROUP II 1.6 MG/KG/DAY DAY 1 2 3 4 5 18956 18971 320. 311. 318. 311. 319. 309. 320. 312. 323. 311. DAY 14 15 16 17 18 18956 18971 353. 325. DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). 6 321. 311. 19 7 333. 317. 20 8 335. 315. 21 9 286. 316. 22 10 331. 318. 11 337. 320. 12 347. 328. 13 359. 328. PROTOCOL 418-014: OR AL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T - 6 2 9 5 .13) TABLE 24 (PAGE 1): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT H DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36 43a 18901 18902 18903 18904 18905 18906 18907 18908 18909 18910 18911 18912 18913 18914 18915 18916 18917 18918 18919 18920 18921 18922 18923 18924 18925 18926 147. 123. c c 133. 144 . 147. 116 . 132. 122. 137. 136. 118 . 152 . 147. 145. 142. 127. 123. 136. 130. 129. 139. 159. 142 . 143 . 146 . 135. 152 . 131. 138. 149. 148. 129. 138 . c 133 . 139. 139. 153. 146 . 138. 148 . 137. 129. 134 . 128. 154 . 127. 160 . 144 . 139. 137. 124. 144 . 128. 130. 150. 144 . 126. 133 . 130. 127. 139. 150. 157. 153. 136. 135. 114 . 132. 126. 132. 146. 133. 165. 144 . 137. 143 . 140. 178. 128 . c c 147. 136. 132. 162. 129. 134 . 129. 161. 104 . 154 . 134 . 106 . 133 . 127 . 132. 138. 132. 162 . 133 . 136. 147. 134. 158. 125. 129. 160. 152. 122 . 133. 135. 143 . 140. 124 . 126 . 188 . 125. 141 . 121. 130. 128. 129. 148. c 159. 147 . 134 . b b 140. b b b b b b b b 118. 124 . b b 135. b 125. c 128. 119 . b d b b 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. 418-014:PAGE B-59 000344 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 I 0 (VEHICLE) MG/KG/DAY DAYS 1- B 8- 15 15- 22 22- 29 29- 36 36 43a 18927 1892B 18929 18930 18931 18932 18933 18934 18935 18936 18937 18938 18939 18940 18941 18942 115. 139. 121 . 151 . 162. 131. 108. 148 . 148. 141 . 136. 138. 137. 146 . 134 . 145. 115. 133. 126 . 165. 162. 134 . 124 . 166 . 146. 145. 145. 148. 145. 155. 131 . 125. 113 . 132. 127. 165. 153. 129. 131. 155. 145. 143. c 140 . 141. 162 . 124 . 129. 111. 133 . 134 . 158. 150. 139. 122 . 157. 141. 138. 125. 141 . 142. 149. 129. 132. 115. 131. 125. 155. 161. 140 . 123 . c 134 . 136. 120. 141. 142. 146. 119. 130. b b b 150. 151. b b b 134 . 80 . b 131. b 138. 126 . 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 s t u d y . c. Spilled feed precluded the calculation of this value. 418-014:PAGE B-60 000345 418-014:PAGE B-i 000346 PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 24 (PAGE 3) : FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT # DOSAGE GROUP II 1 .6 MG/KG/DAY DAYS 1- 8 8- IS 15- 22 22- 29 29- 36 36- 43a 18943 18944 18945 18946 18941 18948 18949 18950 18951 18952 18953 18954 18955 18956 18957 18958 18959 18960 18961 18962 18963 18964 18965 18966 18967 18968 131 . 150. b 129. 182. 156 . 113 . 140. 124 . 145. 134 . 127. b 114 . 142 . 141. 133. 145. 130. 135. 138. 125. 113 . 130. 150 . 139. 126. 137. 149. 126. 140 . 131. 139. 140 . 121 . 144 . 138. 127. 135 . 129. 139. 142 . 139. 154 . 139. 134 . 126 . 120 . 112 . 132. 151 . 157. 123 . 133 . b 131. 147. 121. 134 . 130 . 124 . 137. 135 . 118. b 130. 129. 138. 130 . 142. 132 . 130 . 124 . 125. 115. 126. 147. 133. 126. 145. 136. 130. b 128. 124 . 121 . 125. 135. 132 . 119. b 130. 125 . 124 . 127 . 142 . 129. 134 . 124 . 122. 117 . 126. 152. 136. 121. 144 . b 124 . 131. 131. 123 . Ill . 118. c 136 . 115. 124 . 120 . 136. 120. 127 . 135. 138. 131. 125. 117. 106. 118. 147. 134. 109. 134 . b 122 . 122 . 122 . 122. 106 . 101. 118 . 122. 105. 106 . 114 . 120. 114 . 131 . 125 . 126 . 124 . 116 . 105. 104 . 121. 140 . 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 grou p averages. CD PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 24 (PAGE 4): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DA TA - Fo GENERATION FEMALE RATS RAT # DOSAGE GROUP II 1 .6 MG/KG/DAY DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36- 43a 18969 18970 18971 18972 18973 18974 18975 18976 18977 18978 151. 132. 148. 139. 137. 161 . 152. 130. 134 . 145. 151. 133. 148. 159. 135. 146 . 156. 129. 138 . 145. 149. 129. 134 . 139. 130 . 118. 154 . 137 . 139 . 139. 151. 129. 132 . 142. 129. 124 . 153 . 130. 128. 126. 141. 125. 124. 137. 125. 120. 156. 141. 142. 125. 142. 119. 117. 130. 113. 121. 140. b b 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. 418-014:PAGE B-62 000347 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 25 (PAGE 1): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT (t DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY PREGNANCY STATUS DAYS 0 - 1 1- 2 2- 3 3- 4 4- 5 5-6 18901 P 18902 NP 18903 P 18904 P 18905 NP 18906 P 18907 P 18908 P 18909 P 18910 P 18911 P 18912 P 18913 P 18914 NP 18915 P 18916 P 18917 P 18918 P 18919 P 18920 P 18921 NP 18922 P 18923 P 18924 P 18925 NP 26. 21. 21. 23. 20. 21. 25. 26 . 30. 20. 21. 22. MATING NOT CONFIRMED 28. 31 . 25. 27. 26 . 28 . 25. 22 . 21. 25. 23. 23 . 22. 24 . 21. 27. 26. 26 . 25. 24 . 24 . 22. 24 . 23. 18. 23. 23 . 25. 28 . 27. 21. 17 . 19. 26 . 22. 20. 22. 19. 22 . 25. 23 . 21 . 21. 22 . 22. 18. 8. 17. 23. 23 . 26. 22. 24 . 23. 31. 31. 28. 17. 23 . 23 . 27 . 26. 29. 22. 29. 25 . 22. 21. 24 . 23. 24 . 20. 25. 26 . 24 . 21. 20 . b 22. 18 . 28. 22. 30. 22. 19. 26. 29. 25. 28. 29. 20. 25. 29. 18 . 23 . 18 . 23 . 30. 21. 26. 28 . 23 . 25. 16. 27. 24 . 25. 14 . 21. 24. 27. 24 . 28. 19. 26. 24. 26 . 23 . 22. 20. 28 . 30. 20. 24 . 26 . 23 . 20 . 16. 29. 27. 27. 25. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION 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. 6- 7 26 . 25 . 23. 25. 27. 26. 20. 26 . b 25. 22. 23. 26 . 29. 17. 24 . 28 . 20. 22. 14 . 27. b 26 . 21. 7-8 21. 22. 18. 20. 31. 21. 28. 25. 23. 27. 28. 20. 28. 27. 25. 24. 24. 24 . 20. 17. 28. 19. 37. 22 . 8- 9 26 . 23 . 31. 21. 49. 29. 79. 25. 21. 24. 24. 21. 28. 29. 24 . 26 . 28. 26 . 24 . 17. 28. 22 . 32. 12. 9 - 10 10 - 11 11 - 12 12 - 13 24 . 26. 29. 23 . 22. 1927. 20. 27. 24 . 34 . 24 . 25. 24. 30. 20 6. 39. a 24 . 18. 21. 23. 23 . 27. 25. 22. 22. 27. 25. 22. 14 . 30. 23 . 26 . 23 . 28. 15. 30. 28 . 27. 30. 28. 24 . 28. 34 . 25. 25. 27. 28. 24. 19. 28. 26. 33 . 25. 32. 31. 28. 27. 25. 25. 28. 22 . 16. 32. 23 . 26 . 26 . 30. 27. 17. 29. c 32. 19. 35. 26 . 27. 26. 24 . 29. 29. 26. 18. 28. 20. 24 . 31 28. 27. 22. 29. 25. 36. 15. 418-014:PAGE B-63 000348 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 FEMALE RATS RAT # DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY PREGNANCY STATUS DAYS 0 - 1 1- 2 2- 3 3-4 4- 5 5- 6 18926 P 18927 P 18928 NP 18929 P 18930 NP 18931 P 18932 P 18933 P 18934 P 18935 P 18936 P 18937 P 18938 P 18939 P 18940 P 18941 P 18942 P 19. 25. 25. 26. 26. 26. MATING NOT CONFIRMED 24. 26. 24 . 27. 28. 29. 24 . 27. 26 . 19. 24 . 24. 26 . 25. 25. 27. 27. 29. 14 . 18. 22. 16. 16. 16 . 20. 26 . 24 . 23. 28. 25. 25. 26 . 22 . 23. 26. 31. 23. 24 . 27. 24. 23. 22 . 23 . 26. 27. 27. 29. 25. 25 . 26 . 28 . 15. 23. 25. 26 . 30 . 27. 24 . 22. 24 . 23 . 28. 28. 22. 24 . 30. 27. 13. 23 . 27. 26 . 34 . 24 . 27. 21. 24. 26. 24 . 30. 23. 24 . 30. 26. 14 . 24 . 25. 24 . 23 . 24 . 24 . P = PREGNANT NP NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). 6-7 26. 23 . 22. 32. 24. 25. 25. 38. 21. 24 . 18. 27. 26. 29. 23. 25. 7- 8 23. 29. 26. 30. 31. 20. 29. 32. 19. 28. 26. 26. 25. 29. 27. 24 . 8- 9 22. 23 . 29. 30. 28. 28. 29. 28. 25. 22. 20. 25. 26. 29. 27. 25. 9 - 10 10 - 11 11 - 12 12 - 13 27. 25. 26. 26. 22. 26. 23. 26. 26. 32. 30. 26. 24. 32. 24 . 21. 23. 28. 25 . 33 . 26 . 24 . 32. 27. 31. 25. 24. 26. 25. 28. 24. 29. 28 . 27. 28. 27. 29. 31. 23. 29. 26. 24. 24 . 21. 22. 26. 24 . 30. 23. 26. 31. 25. 32. 24 . 24 . 31. 30. 20. 24. 30. 21. 27. 28. 26 . 418-014:PAGE B-64 000349 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 FEMALE RATS RAT ft DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY PREGNANCY STATUS DAYS 13 - 14 14 - 15 IS - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 21 - 22 22 - 23 23 - 24 24 - 25 18901 P 18902 NP 18903 P 18904 P 18905 NP 18906 P 18907 P 18908 P 18909 P 18910 P 18911 P 18912 P 18913 P 18914 NP 18915 P 18916 P 18917 P 18918 P 18919 P 18920 P 18921 NP 18922 P 18923 P 18924 P 18925 NP 20. 23 . 21. 17. 15. 18. 29. 26. 20. 28. 21. 30. MATING NOT CONFIRMED 28. 15. 30. 31. 28 . 24 . 28 . 19 . 26 . 32. 25. 33 . 28. 23 . 25. 22. 20. 25 . 23. 20. 25. 21. 17. 27 . a 19. 18 . 35. 30 . 39. 25. 21. 15 . 28. 25 . 20. 22. 28 . 17. 26. 16. 27. 24. 22 . 18. 12. 17. 16. 25. 29. 31. 24. 24 . 16. 28. 17. 27. 19. 25. 20. 20. 11. 28 . 25. 31. 28. 25. 31. 28 . 23 . 29 . 27. 23 . 32 . 23 . 26 . 25. 24 . 26. 9. 19. 24 . 26 . 12. 22. 15. 26. 19. 31. 27. 30. 16 . 19. 27. 31. 24 . 15. 29. 21. 26 . 25 . 32 . 27 . 11. 29. 23 . 33. 14 . 25. 11. 30. 27. 28. 27. 20 . 28 . 24 . 19. 21. 20. 19. 36. 18. 24 . 25 . 24 . 30. 16. 27. 22. 20 . 18. 25 . 13. 25. 21. 39. 25. 33. 21. 19. 32. 35 . 24 . 13 . 29. 13 . 23 . 20 . 39. 21 . 16. 27. 16. 36. 17. 29. 16. 22. 19. 19. 22. 18. 21. 29. 23 . 11. 20. 18. 16. 19. 3. 11. 14. 16. 14 . 17. 22 . 4. 23 . 16. 10. 18. 16. 8. 8. 16. 12 . 17. 17. 10. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Value was not recorded. 418-014: PAGE B-65 000350 PROTOCOL 418-014: ORAL (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 DA TA - Fo GENERATION FEMALE RAT ft DOSAGE GROUP I 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 - 24 24 - 25 18926 P 18927 P 18928 NP 18929 P 18930 NP 18931 P 18932 P 18933 P 18934 P 18935 P 18936 P 18937 P 18938 P 18939 P 18940 P 18941 P 18942 P 16. 21 . 26 . 22. 13. 22. MATING NOT CONFIRMED 31. 13. 27 . 10. 19. 20. 27 . 14 . 29. 25. 24 . 19. 25. 23. 32. 27. 31 . 27. 27. 23 . 30 . 26 . 29. 25. 20. 12 . 21 . 28. 13 . 26. 28. 24 . 25. 17. 20. 22 . 23 . 19. 31 . 27. 15 . 17. 27. 20. 28. 19. 27. 23 . 25. 19. 28. 24 . 22. 26 . 31 . 23 . 26 . 31 . 23 . 27 . 27. 12 . 31. 25. 25. 26 . 28. 22 . 24 . 26. 18. 17. 29. 26 . 40. 17 . 22. 23 . 20. 26. 32 . 28. 21. 27 . 19. 20. 23 . 2B . 22 . 19. 32 . 34 . 36 . 19. 32 . 19. 28. 27. 29. 19. 28 . 29. 25. 26 . 36 . 28. 16. 25. 27 . 3. 28. 18. 19. 25. 28 . 7. 29. 13 P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G) . TSDOOO 418-014:PAGE B-66 PROTOCOL 418-014: ORAL (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 DA TA - Fo GENERATION FEMALE RATS RAT # DOSAGE GROUP II PREGNANCY STATUS DAYS 0 - 1 1-2 2- 3 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 18967 P 20. 19. 18. 26. 29. 25. MATING NOT CONFIRMED 19. 22. 21. 20. 21. 22 . 19. 27. 20. 20. 20 . 19. 20. 19. 21. 17 . 20. 21. 23. 20. 23 . 21. 23. 25. 19. 15 . 12. 20. 18. 14 . 21. 20. 17. 17. 23 . 21. 23. 22 . 22 . 17. 20 . 20. 22. 22 . 23 . 19. 22 . 22. 13 . 18 . 18. 17. 20. 22. 20 . 21. 19. 21. 22. 17 . MATING NOT CONFIRMED 14 . 24 . 23 . 1.6 MG/KG/DAY 3- 4 21. 23 . 22. 20. 20. 19. 23 . 20. 24 . 22 . 14 . 23 . 15 . 21. 23 . 18 . 23 . 24 . 12 . 22 . 20. 21. 25. 4- 5 19. 21. 19. 22. 23 . 14 . 21. 22 . 21. 20 . 17. 24 . 15 . 22. 21. 22. 19. 22 . 13 . 23. 23 . 18 . 20. 5-6 7. 29. 23 . 22. 27. 18. 17. 21. 24 . 25. 17. 19. 20. 22. 20 . 19. 22. 23. 17. 22. 20. 17 . 18. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). 6- 7 19. 23. 27 . 21. 22. 17. 21. 25. 23 . 20. 15. 13. 22. 20. 21. 22. 21. 23 . 17. 22 . 23. 26 . 20. 7- 8 21. 26. 19. 23. 26. 22. 27. 26. 21. 21. 15. 19. 23. 26. 20. 23. 25. 21. 15. 23. 24. 18. 30. 8- 9 23 . 26. 19. 25. 27. 13. 20. 19. 25. 21. 17. 18. 19. 25. 25. 21. 27 . 20. 11. 26. 19. 20. 24 . 9 - 10 10 - 11 11 - 12 12 - 13 22. 22. 24 . 22. 29. 29. 24. 28. 23 . 25. 24 . 18. 22 . 27. 21. 23 . 19. 26. 21 . 23 . 26. 24 . 21 . 27. 18 . 25. 23 . 24 . 24. 24 . 25. 20. 21. 22. 26. 25. 17. 16. 21. 25. 24. 25. 26 . 26. 19. 27. 20. 21. 23 . 30. 26. 17. 21. 22 . 25. 25. 12. 22 . 20. 23 . 27. 23 . 27 . 24 . 17. 27. 20. 23. 44 . 22. 29. 15. 20. 24 . 20. 25. 13. 21. 22. 25. 27. 25. 25. 29. 13. 25. 24 . 23. 27. 32. 19. 31. zscooo 418-014:PAGE B-67 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 25 (PAGE 6): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DA TA - Fo GENERATION FEMALE RATS RAT It DOSAGE GROUP II 1.6 MG/KG/DAY PREGNANCY STATUS DAYS 0 - 1 1- 2 2- 3 3- 4 4-5 5- 6 18968 P 18969 P 18970 P 18971 P 18972 P 18973 P 18974 P 18975 NP 18976 P 18977 P 18978 NP 18. 28. 20. 21. 20. 18 . 23 . 22. 18. 19. 17. 22. 25. 18. 17. 18. 18. 20. 22 . 20. 20. 16 . 18. 21. 22. 17. 22 . 17. 18. 22 . 20. 21. 20. 22. 26. 19. 20 . 21. 17. 21 . 19. 24 . 23 . 20. 19. 21. a 27. 23 . 20. 24 . 19. 23 . 24. 20. 23. 25. 21. 20. 24. 18. 24 . 23 . 23 . 25. 25. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Spilled feed precluded the calculation of this value. 6- 7 25. 25. 22. 23. 23 . 17. 25. 23. 27, 26. 21. 7-8 24. 28. 23. 20. 25. 19. 25. 22. 22. 26. 24 . 8- 9 24. 23 . 25 . 20. 24 . 19. 25. 24 . 31. 22. 20. 9-10 21. 27. 21. 23. 25. 23 . 23. 26. 20. 25. 19. 10 - 11 11 - 12 12 - 13 25. 29. 24 . 21. 26. 23 . 24 . 26. 26 . 24. 22. 25 . 26. 23. 22. 27. 21. 29. 23. 27. 27. 10. 27. 31. 23. 20. 30. 25. 27. 19. 28. 28. 12. 418-014:PAGE B-68 000353 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 25 (PAGE 7) : MA TERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT DOSAGE GROU P II 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 - 24 24 - 25 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 18967 p 20 . 24 . 31. 18. S. 17. MATING NOT CONFIRMED 18. 18 . 26. 7. 29. 22. 23 . 14 . 32. 19. 9. 16. 16 . 22 . 26. 15. 25 . 26. 24 . 26. 13. 24 . 16 . 26. 16 . 15 . 18. 20. 14 . 18. 16 . 19. 21 . 25 . 27. 17. 20. 26 . 25. 24 . 23 . 29. 23 . 14 . 23 . 19. 21 . 24 . 17 . 19. 18. 28 . 28 . 19. 20. 25 . 24 . 15. 26 . 26. MATING NOT CONFIRMED 28. 10 . 27. 30 . 15. 26. 32. 32. 9. 30. 29. 26. 25. 7. 10. 27 . 31. 27. 15. 25 . 19. 8. 26 . 22. 30. 29. 27 . 11. 23. 30. 36. 17. 20. 22 . 24. 28. 12. 15 . 23. 27. 25 . 23 . 27. 28. 16 . 27 . 29. 21. 29. 26. 8. 28. 25. 21. 12. 35. 37. 23 . 23 . 16. 13 . 34 . 27. 38. 29. 24 . 27 . 15. 20 . 28. 34. 27. 33. 21. 32 . 13 . 39. 17. 29. 2Q . 20. 34 . 14. 18. 30. 20. 33. 27. 30 . 24 . 15. 15. 24 . 25 . 39. 18. 9. 10 . 13. 14 . 13 . 19. 24. 8. 23 . 7. 7. 15. 18. 18 . 18. 12. 18. 14. 15. 19. 21 . 16 . 16. 12. 5. 14 . 7. 15. 8. 12. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). 418-014:PAGE B-69 000354 PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 25 (PAGE 8): MATERNAL FEED CONSUMPTION VALUES PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RAT (t DOSAGE GROUP II 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 - 24 24 - 25 18968 P 18969 P 18970 P 18971 P 18972 P 18973 P 18974 P 18975 NP 18976 P 18977 P 18978 NP 23. 28. 29. 24 . 29. 29. 24. 20. 23. 20. 27. 31. 31. 21. 42. 20. 21. 17. 24 . 22. 26. 20. 21. 14. 19. 20. 20 . 18. 25. 21. 9. 28. 18 . 29. 27. 31. 25. 37. 24 . 15. 22 . 21. 27. 22. 26. 30. 29. 18. 30. 26. 30. 23. 15. 10. 20. 16. 11. 14 . 24 . 14 . 9. 15. 20. 15. 27. 24 . 21. 25 . 24 . 23 . 18. 26. 31. 31. 19. 21. 28. 29. 28. 16. 26. 18. 3 . 22. 21. 19. 6 . 27. 22. 9. 11. P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). 418-014:PAGE B-70 000355 PROTOCOL 410-014: 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 RATS RATS ASSIGNED TO CROSS- FOSTERING DAYS 1 - 4 4 - 7 1' - 10 10 - 14a RAT # DOSAGE GROUP I b 0 (VEHICLE) MG/KG/DAY SUBSET A 18903 18907 18908 18909 18918C 18920 18924 18929 18934 18936c 18937 18941 18942 47. 51. 48. 48. 82. 63 . 48. 69. 49. 46. 62 . 61 . 85. 109. 117. 104 . 121. 145. 124 . 124 . 127. 108. 117. 123. 104 . 120. 130. 135. 138. 149. 156. 116 .d 134 . 147. 114 . 148. 130. 132. 140. 232. 235. 243 . 207. 264 . 226 . 218. 257. 215. 183 . 250 . 219. 239. rat n DOSAGE GROUP I e 0 (VEHICLE) MG/KG/DAY SUBSET B 18901 18904 18906 18912 18917 18919 18922 18923 18931 18932 18935 18938 96. 71. 90. 64 . 80. 74 . 62. 82. 72. 74 . 60. 70. 135. 143. 128 . 122 . 134 . 130. 117. 149. 130 . 118. 110 . 128. 174. 160. d 168. 154. 147. 147. 145 . 177. 148. 145 . 133. 156 . 255 . 200 . 248. 246 . 240 . 227. 227. 242 . 244 . 203 . 218. 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. 418-014:PAGE B-71 000356 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (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 - 1 0 10 - 14a RAT ft DOSAGE GROUP II b 1.6 MG/KG/DAY SUBSET C 18943 18947 18948 18957 18960 18964 18967 18968 18969 18970 18973 18974 73 . 68. 48 . 51 . 46. 54 . 68. 50. 63. 54. 45 . 31. 123. 141. 240. 113 . 126 . 211 . 95. 106 . 193 . 112 . 128 . 208 . 102. 116. 191.C 98. 126. 182 . 112. 138 . 243. 108. 125. 197. 116 . 128 . 231 . 109. 114 . 191 . 51 . 113 . 182 . NO SURVIVING PUPS ON DAY 4 OF LACTATION RAT tf DOSAGE GROUP II d 1.6 MG/KG/DAY SUBSET D 18946e 18950 18951 18952 18953 18958 18959 18961 18963 18965 18972 18976 18977e 29. 76 . 77 . 124 . 68 . 121 . 135. 227 . 67. 99. 115 . 205 . 74 . 140 . 138. 215 . 59. 104 . 127. 203 . 51 . 87. 119. 202 - 46 . 104 . 135 . 179 . 64 . 119. 134 . 201 . 48 . 114 . 127. 233 . 82 . 116. 133 . 209. 77 . 140 . 148. 258 . 66. 127. 118. 213 . 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. LitterB were not cross-fostered; values excluded from group averages. 418-014:PAGE B-72 000357 PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 26 (PAGE 3): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS RATS ASSIGNED TO PHARMACOKINETIC EVALUATION DAYS 1 - 4 4 - 7 7 - 10 10 - 14 RAT ft DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY 18910 18911 18913 18915 18926 18927 18939 18940 61. 107. 68. 88. 97. 66. 86. 82. 73 . 149. 134. 138. 131. 119. 120. 106 . 114. 148. 140. 216 . 181. 137. 154 . 141. 145. 251. 261. 246. 300. 231. 230. 265. RAT ft DOSAGE GROUP II 1.6 MG/KG/DAY 18956 18971 73. 97. 122. 258. 31. 44 . 57. 77 . DAYS = DAYS OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G) . 418-014PAGE B-73 000358 418-014:PAGE B-74 000359 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 AND SEX - INDIVIDUAL DATA Fo GENERATION FEMALE RATS/F1 GENERATION LITTERS RATS ASSIGNED TO CROSS-]FOSTERING RAT/ LITTER NUMBER LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN NNN TOTAL IMPLANTATIDNS N FOSTER LITTER tt PUPS CROSSFOSTERED N 1 MF NUMBER OF LIVE FOSTER: PUPS AT COMPLETION OF DAY POSTPARTUM 4a 4b 7 14 MF MF MF MF MATERNAL DOSAGE GROUP I C 0 (VEHICLE) MG/KG/DAY SUBSET A 18903 18 0 18 18 18976 15 78 67 55 55 55 18907 19 0 19 20 18963 13 58 57 55 55 55 18908 16 0 16 17 18953 16 4 12 3 12 37 37 37 18909 16 0 16 17 18961 16 88 78 55 55 55 18918d 15 0 15 16 . 18946 17 98 78 55 55 55 18920 17 0 17 17 18952 15 87 85 55 55 55 18924 16 0 16 17 18972 17 98 87 55 55 55 18929 18 0 18 18 18950 16 88 78 55 55 55 18934 16 0 16 18 18959 14 77 76 55 55 55 18936d 12 (1) 0 12 15 18977 12 75 74 64 64 64 18937 14 1 15 18 18958 13 58 58 55 55 55 18941 14 0 14 17 18951 13 85 54 54 53 53 18942 17 0 17 17 18965 18 99 99 55 55 55 MATERNAL DOSAGE GROUP I e 0 (VEHICLE) MG/KG/DAY SUBSET B 18901 18904 18906 18912 18917 18919 18922 18923 18931 18932 18935 18938 16 15 16 15 14 17 15 17(1) 12 16 19 20 0 3 0 1 0 0 0 0 0 0 0 0 16 18 16 16 14 17 15 17 12 16 19 20 17 18923 20 18922 19 18935 16 18938 17 18932 18 18931 18 18904 17 18901 12 18919 18 18917 21 18906 20 18912 16 6 10 6 10 55 55 55 15 78 78 55 55 55 19 8 li 8 10 55 55 55 20 11 9 11 8 55 55 55 16 97 97 55 55 55 12 75 75 55 55 55 15 5 10 5 10 55 55 55 16 6 10 69 55 55 55 17 7 10 7 10 55 55 55 14 68 68 55 55 55 16 79 79 55 55 55 15 13 2 13 2 82 82 82 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. 21 MF 55 55 37 55 55 55 55 55 55 64 55 53 55 55 55 55 55 55 55 55 55 55 55 55 82 418-014:PAGE B-75 0003G0 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 27 (PAGE 2): NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA Fo GENERATION FEMALE RATS/FI GENERATION LITTERS RATS ASSIGNED TO CROSS-FOSTERING RAT/ LITTER NUMBER LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN NNN TOTKti IMPLAN TAT INS N1 FOSTER LITTER U PUPS CROSSFOSTERED N l MF NUMBER OF LIVE FOSTER PUPS AT COMPLETION OF DAY POSTPARTUM 4a 4b 7 14 MF MF MF MF MATERNAL :DOSAGE GROUP I]: c 1.6 MG/KG/DAY SUBSET iC 18943 18947 18948 18957 18960 18964 18967 18968 18969 18970 18973 18974 16 15 17 15 15 10 15 15 17 13 16(1) 14 0 0 0 0 0 0 0 0 0 0 0 0 16 15 17 15 15 10 15 15 17 13 16 14 16 ` 16 19 15 17 13 ' 17 16 19 13 16 17 18967 18974 18969 18970 18973 18968 18943 18964 18948 18957 18960 18947 15 5 10 5 10 55 55 55 14 86 86 55 55 55 17 11 6 62 62 62 62 13 49 38 37 37 37 15 5 10 38 37 37 37 15 87 77 55 55 55 16 97 97 55 55 55 10 28 18 18 18 18 17 12 5 12 5 55 55 55 15 96 95 55 55 55 15 10 5 10 4 64 64 64 15 69 -- -* -- -- 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 17 16 13 15 16 13 14 16 13 18 17 15 12 0 17 0 16 0 13 2 17 0 16 13 0 14 0 16 0 13 0 18 0 17 0 15 0 12 17 17 ' 13 : 17 18 14 15 16 13 18 ' 18 15 16 18918 18929 18941 18920 18908 18937 18934 18909 18907 18942 18924 18903 18936 15 98 31 31 21 21 18 8 10 8 10 55 55 55 14 86 86 55 55 55 17 89 89 55 55 55 16 88 88 55 55 55 14 68 68 55 55 55 16 97 96 55 55 55 16 5 11 5 11 55 55 55 19 11 8 11 a 55 55 55 17 6 11 6 li 55 55 55 16 79 79 55 55 55 18 8 10 7 10 55 55 55 12 74 -- -- -- -' 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-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. Litter were not cross-fostered; values excluded from group averages. 21 MF 55 55 62 37 37 55 55 18 55 55 64 -- 21 55 55 55 55 55 55 55 55 55 55 55 -- 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/FI GENERATION LITTERS RATS ASSIGNED TO PHARMACOKINETIC EVALUATION RAT/ LITTER NUMBER LITTER DELIVERED LIVE STILL- TOTAL BORN BORN BORN NNN NUMBER OF LIVE PUPS AT COMPLETION OF DAY POSTPARTUM 1 4 7 14 MF MF MF MF MATERNAL DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY 18910 18911 18913 18915 18926 18927 18939 18940 50 5 16 0 16 18 0 18 15 2 17 12 0 12 17 0 17 15 0 15 16 0 16 32 11 5 16 2 4 11 66 89 78 10 6 32 11 5 15 2 4 11 66 88 78 10 6 32 11 4 14 2 4 11 66 88 78 10 6 32 11 4 14 2 4 11 66 87 7a 10 6 MATERNAL DOSAGE GROUP II 1.6 MG/KG/DAY 18956 18971 12 1 13 123 57 -1 57 -1 57 -1 57 1 M = MALE F = FEMALE IMPLANTATIONS N TOTAL 8 16 19 17 13 17 15 18 16 3 418-014:PAGE B-76 000361 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 (MINUTES) PER PUP PER LITTER - INDIVIDUAL DA TA Fo GENERATION FEMALE RATS RATS ASSIGNED TO CROSS-FOSTERING RAT/ LITTER NUMBER DURATION OF GESTATION (DAYS) a N DURATION OF GESTATION (WHOLE DAYS) N AVERAGE DURATION OF DELIVERY PER PUP (MINUTES) b MATERNAL DOSAGE GROUP I c 0 (VEHICLE) MG/KG/DAY SUBSET A 18903 18907 18908 18909 18918d 18920 18924 18929 18934 18936d 18937 18941 18942 21.6 21.5 21.7 22.5 22.6 21.6 21.6 22.4 21.5 21.7 22.5 22.4 21.8 22 16.9 22 14.5 22 8.5 23 15.5 23 7.2 22 9.1 22 6.5 23 6.7 22 5.1 22 25.1 23 23.2 23 15.3 22 9.5 MATERNAL DOSAGE GROUP I e 0 (VEHICLE) MG/KG/DAY SUBSET B 18901 18904 18906 18912 18917 18919 18922 18923 18931 18932 18935 18938 22.3 22.6 21.6 21.6 22.5 21.7 21.6 22.8 21.6 22.5 21.6 21.6 23 21.8 23 11.4 22 8.8 22 12.7 23 13.4 22 9.1 22 13.3 23 11 .I t 22 12.4 23 7.2 22 7.4 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-1 pups. c. Vehicle control grou p dams cross-fostered with 1.6 mg/kg/day dosage gr ou p 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. 418-014:PAGE B-77 000362 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 - INDIVIDUAL DA TA Fo GENERATION FEMALE RATS RATS ASSIGNED TO CROSS-FOSTERING RAT/ LITTER NUMBER DURATION OF GESTATION (DAYS) a N DURATION OF GESTATION (WHOLE DAYS) N AVERAGE DURATION OF DELIVERY PER PUP (MINUTES) b MATERNAL DOSAGE GROU P II c 1.6 MG/KG/DAY SUBSET C 18943 18947 18948 18957 18960 18964 18967 18968 18969 18970 18973 18974 21.5 21.5 21.6 21.6 21.6 2 i .e 21.5 21.7 21.6 21.7 21.5 21.5 22 4.9 22 10.8 22 11.2 22 11. 1 22 3.1 22 9.9 22 4.6 22 9.5 22 11.2 22 8.3 22 11.0 22 8.6 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 21.5 21.5 21.6 21.5 21.7 21.7 21.5 21.5 21.6 21.4 21.7 21.1 21.6 22 1 2 .3f 22 8.3 22 11.2 22 13.2 22 14.1 22 5.5 22 9.2 22 7.9 22 11.2 22 6.4 22 4.4 22 13.8 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 pups. c. 1.6 mg/kg/day dosage group dams cros3-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross -fostered with vehicle control grou p litters. e. Litters were not cross-fostered; values excluded from group averages. f. Partuition period only partially observed. 418-014:PAGE B-78 000363 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 DE LIVERY (MINUTES) PER PUP PER LITTER - INDIVIDUAL DATA Fo GENERATION FEMALE RATS RATS ASSIGNED TO PHARMACOKINETIC EVALUATION RAT/ LITTER NUMBER DURATION OF GESTATION (DAYS) a N DURATION OF GESTATION (WHOLE DAYS) N AVERAGE DURATION OF DELIVERY PER PUP (MINUTES) b MATERNAL DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY 18910 18911 18913 18915 18926 18927 18939 18940 21.7 21.8 22.4 21.6 22.4 22.3 21.9 21.7 22 14.8 22 14.9 23 6.1 22 30.4 23 5.2 23 9.5 22 9.6 22 9.2d MATERNAL DOSAGE GROUP II 1.6 MG/KG/DAY 18956 18971 22.5 C 23 8.2 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 pups. c. Partuition period not observed. d. Partuition period only partially observed. 418-014:PAGE B-79 000364 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 29 (PAGE 1) : PUP BODY WEIGHT LITTER AVERAGES FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION LITTERS RATS ASSIGNED TO CROSS-FOSTERING RAT/ FOSTER LITTER DAY 1 M FT DAY 4a M FT DAY 4b MFT DAY 7 MF T DAY 14 MF T MATERNAL !DOSAGE GROUP I c 0 (VEHICLE) MG/KG/DAY SUBSET A 18903 18907 18908 18909 1891Bd 18920 18924 18929 18934 18936d 18937 18941 18942 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 6.1 5.8 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 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 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 5.6 S .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 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 5.7 5.5 5.6 7.5 6.7 7.X 7.5 7.3 7.4 12.5 12.0 12.3 25.9 25.3 25.6 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 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 6.2 5.8 6.1 8.7 7.7 8.3 a.e 7.7 8.2 13.1 12.0 12.7 25.1 23.0 24.2 6.0 5.5 5.7 8.0 7.3 7.6 8.0 7.5 7.7 11.9 11.5 11.7 24.9 25.2 25.1 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 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 MATERNAL DOSAGE GROUP I e 0 (VEHICLE) MG/KG/DAY SUBSET B 18901 18904 18906 18912 18917 18919 18922 18923 18931 18932 18935 18938 6.7 6.2 6.4 5.9 5.6 5.7 5.8 5.5 5.6 5.5 5.2 5.4 7.0 6.7 6.9 6.4 6.3 6.4 6.8 6.4 6.5 7.0 6.7 6.8 6.4 6.0 6.2 6.8 6.2 6.4 6.3 5.8 6.0 6.1 5.8 6 .1 9.4 8.6 8.9 7.6 7.0 7.3 8.2 7.6 7.9 6.8 6.2 6.6 8.7 8.4 8.5 9.5 9.3 9.4 8.9 8.5 8.6 9.6 8.9 9.2 8.3 8.0 8.1 9.1 8.6 8.8 8.4 7.7 8.0 8.5 8 .4 8 .5 9.5 8.5 9.0 15.6 14.3 15.0 31.7 30.1 30.9 8.0 7.4 7.7 14.2 12.6 13.4 28.9 26.5 27.7 8.4 8.1 8.3 13.5 13.4 13.4 28.7 28.3 28.5 7.0 6.3 6.6 12.5 12.7 12.6 29.8 27.5 28.6 8.6 8.4 8.5 13.8 13.6 13.7 28.9 28.4 28.6 9.3 9.3 9.3 15.6 15.1 15.4 30.9 29.9 30.4 8.9 8.9 8.9 15.5 15.1 15.3 30.8 31.0 30.9 9.8 8.8 9.3 16.1 14.3 15.2 30.5 28.1 29.3 8.2 8.2 8.2 13.8 13.8 13.8 30.6 30.5 30.6 9.2 8.7 8.9 14.5 13.9 14.2 27.4 26.9 27.2 8.5 8.1 8.3 13.5 13.0 13.2 26.2 25.9 26.0 8 .7 8.4 8.6 13.8 13.8 13.8 29.2 29.0 29.1 M = MALE F = FEMALE T = TOTAL (SUM OF PUP WEIGHTS/NUMBER OF LIVE PUPS) DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. a. Preculling 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 gr ou p averages. e. Vehicle control group dams cross-fostered with vehicle control litters. DAY 21 MF T 41.1 44.2 42.1 42.7 50.0 41.4 41.2 47.2 42.0 39.9 40.3 44.5 41.7 38.2 42.6 39.9 40.5 49.9 40.5 40.9 43.8 40.2 36.6 41.0 45.4 41.6 39.7 43.4 40.6 41.6 50.0 40.9 41.0 45.5 41.1 38.6 40.7 44.8 41.6 50.0 44.4 49.9 48.6 45.3 49.0 48.7 50.4 48.9 43.7 42.3 44.5 46.4 41.5 47.9 44.7 45.7 47.8 48.4 46.1 47.7 43.5 41.5 43.8 48.2 43.0 48.9 46.7 45.5 48.4 48.5 48.3 48.3 43.6 41.9 44.3 418-014.PAGE B-80 000365 PROTOCOL 4X8-014: 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 - FI GENERATION LITTERS RATS ASSIGNED TO CROSS-FOSTERING RAT/ FOSTER LITTER # DAY 1 MF T DAY 4a MFT DAY 4b M FT DAY 1 MF T DAY 14 MF T MATERNAL DOSAGE GROUP II 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 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 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 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 5.4 5.3 5.4 6.8 6.2 6.3 6.8 6.4 6.5 10.3 9.3 9.6 22.9 20.4 21.2 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 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 6.6 6.2 6.3 10.4 8.7 a.9 10.4 8.7 8.9 15.4 13.4 13.6 30.4 26.6 27.1 5.6 5.6 5.6 7.4 7.5 7.4 7.7 7.5 7.6 12.7 11.9 12.3 27.0 25.6 26.3 5.7 5.4 5.6 7.1 5.6 6.6 7.4 5.6 6.5 12.6 9.4 11.0 25.1 20.0 22.5 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 5.7 5.3 5.4 NO SURVIVING PUPS MATERNAL DOSAGE GROUP II d L.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 5.4 5.8 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 8.1 7.3 7.7 8.4 7.6 8.0 13.3 12.3 12.8 26.9 25.4 26.1 8.8 8.4 8.6 9.0 8.4 8.7 12.6 11.2 11.9 24.2 21.6 22.9 8.2 7.4 7.8 8.7 7.6 8.2 14.8 13.3 14.0 30.7 28.1 29.4 8.2 8.2 8.2 8.6 8.5 8.6 12.7 11.9 12.3 25.4 24.8 25.1 9.2 8.1 8.6 9.1 8.4 8.7 12.9 12.3 12.6 24.7 24.0 24.4 8.2 7.6 8.0 8.0 7.7 7.8 13.0 12.5 12.7 27.1 26.2 26.6 8.3 7.7 7.9 8.3 8.5 8.4 13.3 13.1 13.2 26.3 25.7 26.0 7.1 6.4 6.8 7.3 6.6 6.9 12.2 11.7 12.0 26.8 25.9 26.4 8.1 7.6 7.8 8.2 7.7 8.0 12.7 12.3 12.5 25.6 24.9 25.2 9.0 8.1 8.5 9.0 8.3 8.7 14.8 14.0 14.4 31.4 29.9 30.6 7.4 7.1 7.2 7.6 7.2 7.4 12.8 12.5 12.6 26.2 24.8 25.5 NO SURVIVING PUPS M = MALE F = FEMALE T = TOTAL (SUM OF PUP WEIGHTS/NUMBER OF LIVE PUPS) DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. a. Preculling b. Postculling 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. DAY 21 MF T 43.1 43.1 36.8 39.7 35.9 33.9 47.1 50.7 40.5 39.9 36.6 42.0 39.7 33.6 39.8 32.4 35.0 41.1 42.9 38.2 33.0 36.6 42.5 41.4 36.0 39.8 33.4 34.4 44.1 43.8 39.4 36.4 36.6 48.8 44.5 39.4 47. 1 41.6 40.2 42.8 42.0 43.2 44.0 46.1 41.7 36.3 39.7 36.5 43.4 40.3 39.5 42.1 40.1 41.1 42.3 43.7 39.6 44.6 42.1 38.0 45.3 41.0 39.8 42.5 41.1 42.5 43.1 44.9 40.6 418-014.PAGE B-81 000366 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 29 (PAGE 3) : PUP BODY WEIGHT LITTER AVERAGES FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION LITTERS RATS ASSIGNED TO PHARMACOKINETIC EVALUATION RAT/ LITTER NUMBER DAY 1 MF T DAY 4 MF T DAY 7 MF T DAY 14 MF T MATERNAL DOSAGE GROUP i 0 (VEHICLE) MG/KG/DAY 18910 18911 18913 18915 18926 18927 18939 18940 7.4 7.1 7.3 a a a 17.5 16.4 17.1 33.8 32.1 33.1 6.6 6.5 6.6 8.9 8.4 8.8 12.4 12.5 12.4 23.7 24.1 23.8 6.2 5.7 6.2 8.0 7.6 a.o 10.9 10.6 10.9 21.2 19.6 21.0 6.4 6.1 6.2 a a a 12.6 13.0 12.9 24.5 25.3 25.0 7.0 7.0 7.0 11.9 12.4 12.2 15.9 15.6 15.8 30.0 29.4 29.7 6.6 6.1 6.3 8.8 8.2 8.5 12.3 10.8 11.5 22.4 20.6 21.6 6.4 6.0 6.2 a a a 12.9 12.5 12.7 23.6 22.6 23.1 6.2 5.8 6.1 8.5 8.3 8.4 10.8 10.8 10.8 1 9 .B 18.8 19.4 MATERNAL DOSAGE GROUP II 1.6 MG/KG/DAY 18956 18971 6.4 6.0 6.2 -- 7.6 7.6 9.3 8.8 9.0 13.1 12.1 12.5 24.2 23.4 23.8 -- 9.1 9.1 -- 9.2 9.2 -- 11.7 11.7 M = MALE F = FEMALE T = TOTA L (SUM, OF PUP WEIGHTS/NUMBER OF LIVE PUPS) DAY = DAY POSTPARTUM ALL WEIGHTS WERE RECORDED IN GRAMS (G)-P: MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS, a. Values were not recorded. 418-014:PAGE B-82 000367 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 30 (PAGE 1): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS RATS ASSIGNED TO CROSS-FOSTERING PUP H 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 i a 0 (VEHICLE) MG/KG/DAY SUBSET A POSTPARTUM DAY 1 18903 18907 18908 18909 18918b 18920 18924 18929 18934 18936b 18937 18941 18942 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 5.8 5.5 6.2 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 5.2 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 5.2 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 5.3 5 . 8 5 . 9 5.7 .0 5.9 5.8 5.0 5.5 5 . 7 5.4 5.5 5.4 5.0 5.7 5.5 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.S 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 6.4 6 . 3 5.9 6 . 6 6 . 0 6.2 6 . 4 5 . 9 5.7 5.9 5.9 5.8 6 . 2 5 . 9 6.0 6.2 5 . 8 5.6 5.1 5 . 5 5.7 5.5 5.7 5.6 5.6 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 5.3 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 5.4 5.0 RAT/ LITTER D 18901 18904 18906 18912 18917 18919 18922 18923 18931 18932 18935 18938 MATERNAL DOSAGE GROUP I P 0 (VEHICLE) MG/KG/DAY SUBSET B POSTPARTUM DAY 1 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 5.1 6.1 5 . 8 5 . 7 6 . 1 5.0 5 . 9 6 . 1 5.6 5.6 5.4 4 . 9 5.7 5.1 5.7 5.5 5.5 5.8 5.8 5.3 5.7 5.2 5.9 5.8 5.2 5.6 5.6 5.4 5.6 5.6 5.3 5.1 5.6 5.1 5.5 5.2 4.8 4.8 5.0 5.6 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 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 6.1 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 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 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. 418-014:PAGE B-83 000368 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 - FI GENERATION PUPS RATS ASSIGNED TO CROSS-FOSTERING PUP # 12 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 18948 18957 18960 18964 18967 18968 18969 18970 18973 18974 5.7 5.8 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 5.6 5.4 6.1 5.9 5.9 5.8 5.8 6.2 5.4 5.7 5.6 5.7 5.5 5.7 6.1 5.5 5.9 5.8 5.9 5.8 5.5 6.1 5.8 6.2 5.6 5.8 4.4 5.5 5.9 5.2 6.2 6.7 6.2 5.6 6.1 5.8 S .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 5.4 5.1 5.2 7.3 6 .0 6 .3 6 .1 6.0 5.6 6 . 6 6 .9 6.1 5.9 5.9 5 . 6 5 .8 5.6 6 . 0 5.7 5 . 5 5.3 5 . 5 5.6 5 .8 5.6 5.7 5.6 5.4 5.6 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 5.8 5.9 5.3 5 .2 6 .1 5.4 5.8 5.4 5.6 5.7 5.4 5 . 5 5.2 5 .6 5 . 5 5.6 5 . 5 5.9 5.9 5.6 5 .5 5 .4 5.2 5 .3 5.4 5.3 4.6 5.4 5 .3 5.6 RAT/ LITTER It MATERNAL DOSAGE GROUP II b 1 . 6 MG/KG/DAY SUBSET D POSTPARTUM DAY 1 18946c 18950 18951 18952 18953 18958 18959 18961 18963 18965 18972 18976 18977c 7.0 6 .5 8.4 5.6 6 .9 7.1 6.1 6.1 6 .1 6.4 6.4 6.2 6.7 7.2 6.7 7.7 6.3 7.1 7.4 6.2 6.2 5.6 6.0 6 .1 6.0 7 .1 6.6 7.0 7.2 6 .6 6.7 6.8 6.0 6.9 5.6 6 .1 6 .2 6 .1 6 .1 6.8 7 .0 7.8 6.4 7.3 7.1 6 -0 6.1 5 .7 5 .9 6 .4 5.9 6 .S 6.7 7.9 7.9 6.5 7 .2 7.3 6.4 6.2 5.9 5 .8 6 .4 6 .0 6 .4 7 .0 7 .3 7.8 6 .8 6.9 7.2 6 .0 65 6 .0 5 .9 6 .7 6 .3 7 .0 7.3 7 .3 7.3 6 .3 6 .7 6.3 5.9 5.8 5 .8 5 .5 6 .3 6 .3 7 .0 6.6 7.1 7.9 6.4 7.3 5.5 5.8 6.1 5.5 5.9 5.7 6.5 6 .1 6 .6 6.3 7.4 5.7 6.9 6.4 6.0 6.4 6 .0 6 .1 6 .2 5 .8 5 .9 6 .4 7 .0 7.6 6.1 6.1 6.8 5.5 6.3 5 .7 5 .4 5.9 5.9 6 .3 6.6 6.4 7.6 6.1 7.1 6.7 5.3 6.3 5.6 5.2 6 .1 6.2 6 .1 6.1 6 .6 7.8 6.1 6.8 6.5 5.9 6.4 5 .2 5.3 5 .7 5 .9 FD 1 6.2 6.0 7.0 5.7 6.4 7.2 5.8 6.2 5.8 6.0 5.4 5.4 6.4 6.4 7.7 5.9 6.1 6.0 5.5 5.4 5.5 5 .9 5.6 5 .7 6.7 5 .7 6 .2 6.3 6.1 5.5 5 .6 5.8 5.9 6 .0 d 6 .2 5.8 6.6 4.1 6.0 5 .5 5 .9 5.6 6 .1 d 6.1 5.8 5 .0 6 .0 5.9 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 litterB. 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. d. Value was not recorded. IB 6.6 5.4 5 .9 19 20 418-014:PAGE B-84 000369 PROTOCOL 418-014: ORAL (GAVAGE) 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 - FI 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 18 19 20 RAT/ LITTER tt MATERNAL DOSAGE GROUP I a 0 (VEHICLE) MG/KG/DAY SUBSET A POSTPARTUM DAY 4 PRECULLING 18903 18907 18908 18909 18918b 18920 18924 18929 18934 18936b 18937 18941 18942 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 e .o 6.7 8.8 7.3 8.4 7.7 8.1 5.3 8.2 7.4 6.7 8.1 FD 4 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 7.6 7.7 5.3 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 10.S 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 8.7 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 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 4 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 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 8.4 9.2 9.2 8.9 8.4 8.2 8.6 7.2 8.1 7.7 7.8 FM 2 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 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 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 7.3 7.6 RAT/ LITTER It MATERNAL DOSAGE GROUP I c 0 (VEHICLE) MG/KG/DAY SUBSET B POSTPARTUM DAY 4 PRECULLING 18901 18904 18906 18912 18917 18919 18922 18923 18931 18932 18935 18938 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 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 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 8.0 7.6 FD 4 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 6.0 6.3 4.7 FM 4 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 10.2 9.6 8.7 9.4 9.0 10.1 9.3 9.5 9.1 9.5 9.3 9.1 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 9.9 9.9 8.4 9.3 10.6 9.5 9.4 8.1 9.3 6.6 9.4 9.7 9.2 7.6 9.2 FM 3 8.3 7.7 8.6 8.2 8.3 8.6 8.2 8.5 7.5 7.9 8.3 8.2 8.0 7.2 7.6 8.7 8.2 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 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 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-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 gr o u p litters. b. Litters were not cross-fostered; values excluded from group averages. c. Vehicle control group dams cross-fostered with vehicle control litters. 418-014:PAGE B-85 000370 PROTOCOL 418-014: OR AL (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 - FI 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 18 19 RAT/ LITTER # MATERNAL DOSAGE GROUP1 II a 1.6 MG/KG/DAY SUBSET C POSTPARTUM DAY 4 PRECULLING 18943 18947 18948 18957 18960 18964 18967 18968 18969 18970 18973 18974 7.1 7.6 6.1 8.8 6.6 6.9 9.0 10.4 7.4 7.6 6.1 MD 2 7.8 8.0 7.1 7.0 7.3 6.6 8.6 MD 2 7.9 7.1 8.7 MD 2 8.7 7.7 6.1 7.1 6.5 6.6 7.8 9.3 6.8 7.1 7.7 MD 2 7.5 7.4 7.1 MM 3 MM 4 6.5 6.7 8.3 7.2 7.8 7.8 MD 2 8.8 7.0 4.4 6.9 MM 4 7.1 8.3 9.4 7.6 6.2 7.3 MD 2 8.2 7.9 7.2 6.8 6.0 6.2 9.0 9.4 7.6 6.9 6.7 MD 2 8.6 7.9 MM 4 6.9 6.4 MM 3 7.7 9.1 6.8 7.7 7.0 FM 4 8.8 7.4 MM 3 7.3 7.4 6.3 6.6 8.5 7.3 6.1 7.8 FM 3 7.3 6.3 MD 2 8.7 6.5 6.0 8.1 7.6 7.4 7.7 7.8 FD 2 8.2 7.6 MD 2 7.7 6.5 7.4 7.5 8.0 7.5 6.9 6.8 FD 2 8.2 7.7 MD 2 7.6 6.6 6.3 7.3 6.9 4.3 6.2 FD 2 8.5 7.0 6.5 7.7 4.6 6.1 7.7 8.2 3.9 7.5 FD 2 7.6 7.1 5.1 FM 3 5.2 7.0 7.9 7.9 6.0 7.2 FD 2 6.9 6.7 FD 3 FM 4 7.6 7.6 7.5 6.7 7.8 FD 2 8.4 FD 3 FM 4 6.6 6.1 7.8 FM 3 FD 2 FD 2 FM 3 6.0 8.2 FM 3 6.3 RAT/ LITTER tt MATERNAL DOSAGE GROUP II b 1.6 MG/KG/DAY SUBSET D POSTPARTUM DAY 4 PRECULLING 18946c 18950 18951 18952 18953 18958 18959 18961 18963 18965 18972 18976 18977c 8.7 7.3 8.8 8.7 8.0 9.3 8.4 7.8 7.1 8.2 8.6 8.0 MD 3 6.0 7.9 8.9 6.3 8.7 7.8 8.0 8.9 7.0 8.5 8.5 8.0 MD 2 4.3 7.5 8.0 8.9 5.5 10.0 B.O 8.4 6.8 8.4 9.7 6.8 MD 2 MD 4 8.3 8.6 8.5 8.8 8.9 8.1 8.0 7.4 7.3 9.1 7.0 MD 2 MD 4 8.7 8.9 8.3 8.3 9.5 7 .a 8.5 7.0 8.1 8.7 7.7 MD 2 MM 3 7.8 9.6 7.8 8.4 9.5 8.5 6.0 7.3 8.0 9.1 7.3 MD 2 MD 2 8.3 8.8 8.9 9.1 7.8 8.0 9.1 6.5 7.5 9.1 7.3 MD 2 MD 2 9.1 8.6 8.4 8.9 8.1 8.3 7.1 6.7 8.2 8.3 MD 2 FD 2 MD 2 6.7 8.9 7.1 8.8 8.7 8.8 6.8 8.0 8.2 8.5 7.6 FD 2 5.2 7.2 8.3 6.6 7.4 8.4 8.0 8.5 7.1 7.4 7.4 6.7 FD 2 FM 3 6.7 8.1 7.8 8.0 8.8 7.3 7.6 7.3 7.3 8.1 6.6 FD 2 FD 2 8.8 7.5 7.5 9.0 6.9 7.3 7.5 6.5 6.3 8.2 6.4 FD 1 FD 2 7.4 8.7 7.6 7.8 8.5 8.4 8.1 5.6 7.2 7.4 7.0 FD 2 7.4 8.6 7.7 7.8 7.7 7.8 8.3 6.4 7.2 7.2 7.4 FD 2 7.3 7.7 8.3 7.0 8.3 6.9 7.6 8.6 7.4 FD 2 6.3 7.4 8.9 FM 3 7.7 6.8 8.0 9.0 7.4 FD 2 7.8 7.4 6.1 8.6 7.0 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. 418-014:PAGE B-86 000371 PROTOCOL 418-014: OR AL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 30 (PAGE 5): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - FI 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 18 19 20 RAT/ LITTER # MATERNAL DOSAGE GROUP I fi 0 (VEHICLE) MG/KG/DAY SUBSET A POSTPARTUM DAY 4 POSTCULLING 18903 18907 18908 18909 18918b 18920 18924 18929 18934 18936b 18937 18941 18942 MC 4 8.0 7.0 7.6 10.5 7.2 MC 4 MC 4 8.0 8.4 8.1 8.7 MC 4 5.9 6.7 6.7 7.7 MC 4 MC 4 8.4 MC 4 7.5 9.2 8.2 8.2 7.6 5.7 8.8 7.5 MC 4 9.7 6.9 7.3 8.6 MC 4 MC 4 8.3 8.2 7.9 6.5 7.3 MM 4 7.9 9.5 7.4 7.5 8.0 7.1 8.9 7.7 7.1 MC 4 6.5 8.4 7.4 7.6 MC 4 7.1 MC 4 7. S 8.3 8.4 7.7 9.0 MC 4 6.0 7.7 7.6 MC 4 10.1 MC 4 MC 4 7.9 7.0 8.2 7.4 MM 3 MC 4 MM 3 8.1 FC 4 7.8 10.4 7.7 7.0 8.7 MC 4 8.6 FC 4 MM 3 7.1 6.8 FC 4 7.2 MD 4 MM 2 MC 4 MM 3 MM 3 FC 4 7.2 7.7 MD 2 6.8 FC 4 8.2 FC 4 8.0 MM 2 7.0 7.2 7.3 7.1 8.1 7.4 9.3 6.7 6.5 7.4 FC 4 8.1 FC 4 7.6 7.0 FC 4 7.4 7.7 7.1 6.1 6.9 FC 4 6.7 6.5 7.0 9.0 7.1 7.7 FC 4 7.5 7.8 7.7 8.7 PC 4 6.0 FC 4 FC 4 7.5 9.3 7.1 7.6 FC 4 7.1 FM 2 FC 4 3.6 FC 4 5.8 FD 4 FC 4 FC 4 FC 4 6.2 6.8 6.4 5.9 FC 4 FD 2 7.2 4.6 8.0 FC 4 FC 4 FM 3 FC 4 8.2 FD 2 FC 4 FM 3 7.3 FC 4 9.5 FD 2 7.3 7.4 7.9 6.5 7.8 9.0 FC 4 7.9 6.9 8. 7 FD 4 FC 4 7.6 RAT/ LITTER # MATERNAL DOSAGE GROUP I f 0 (VEHICLE) MG/KG/DAY SUBSET B POSTPARTUM DAY 4 POSTCULLING 18901 18904 18906 18912 18917 18919 18922 18923 18931 18932 18935 18938 MC 4 MC 4 MC 4 MC 4 9.3 MC 4 8.1 9.9 8.3 9.3 9.0 8.5 8.8 MC 4 8.3 7.2 9.0 MC 4 10.1 9.9 7.7 MC 4 8.7 9.5 9.5 9.2 8.8 MC 4 MC 4 8.7 8.8 MC 4 8.6 9.8 MC 4 8.5 10.7 7.8 MC 4 7.0 MC 4 9.4 8.7 9.3 MC 4 9.0 8.8 MC 4 9.5 ,7.9 MC 4 5.5 7.1 9.0 8.7 1. .6 8.3 8-8 8.7 MC 4 9.2 7.7 7.7 MC 4 9.1 10.1 FC 4 9.5 MC 4 9.0 MC 4 8.1 8.3 7.2 9.0 6.9 8.7 9.3 FC 4 FC 4 8.2 8.5 7.5 MC 4 8.3 FC 4 8.4 MC 4 MC 4 9.5 FC 4 FC 4 8.5 9.2 8.7 9.1 8.6 FC 4 FC 4 MC 4 MC 4 9.1 8.7 9.3 FC 4 8.5 FC 4 8.1 FC 4 8.6 FC 4 7.4 8.9 9.5 9.2 8.6 7.9 FC 4 8.2 MC 4 FC 4 7.5 8.8 MC 4 FC 4 9.3 9.4 FC 4 FC 4 8.5 FC 4 9.1 FC 4 7.7 FC 4 6.8 8.6 9.1 9.5 FC 4 FC 4 FC 4 FC 4 8.6 FC 4 FC 4 7.9 FC 4 8.1 FC 4 9.2 FC 4 8.7 FC 4 MC 4 FC 4 6.8 7.6 6.8 7.7 FC 4 7.6 FC 4 FC 4 8.7 8.2 7.9 6.3 7.8 FC 4 FC 4 7.9 9.2 7.6 7.5 8.5 9.3 8.4 6.7 8.5 FM 3 8.7 7.3 FC 4 FC 4 8. 2 FC 4 6.3 FD 4 4.7 FM 4 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 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. 418-014:PAGE B-87 000372 PROTOCOL 418-014: ORAL (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 - FI GENERATION PUPS RATS ASSIGNED TO CROSS-FOSTERING PUP # 12 3 4 S 6 7 8 9 10 11 12 13 14 15 16 17 18 19 RAT/ LITTER # MATERNAL DOSAGE GROUP II a 1.6 MG/KG/DAY SUBSET C POSTPARTUM DAY 4 POSTCULLING 18943 18947 18948 18957 18960 18964 18967 18968 18969 18970 18973 18974 7.1 7.6 6.1 8.6 6.6 6.9 9.0 10.4 MC 4 MC 4 MC 4 MD 2 7.8 8.0 7.1 7.0 7.3 6.6 8.6 MD 2 7.9 MC 4 8.7 MD 2 8.7 MC 4 6.1 7.1 6.5 MC 4 MC 4 9.3 MC 4 7.1 MC 4 MD 2 7.5 MC 4 7.1 MM 3 MM 4 MC 4 MC 4 8.3 MC 4 7.8 MC 4 MD 2 8.8 FC 4 MC 4 7.9 4.4- 7.2 FC 4 6.8 MM 4: 6.0 7.1 6.2 8.3 MC 4 9.4 9.4 MC 4 7.6 MC 4F; 6.9 7.31, 6.7 MD 2- ; MD 2 FC 4 7.9 MM 4 6.9 6.4 MM 3 7.7 9.1 MC 4 7.7 MC 4 FM 4 8.8 7.4 MM 3 7.3 7.4 6.3 MC 4 8.5 7.3 MC 4 7.8 FM 3 7.3 FC 4 MD 2 8.7 6.5 FC 4 8.1 7.6 MC 4 7.7 7.8 FD 2 8.2 7.6 MD 2 7.7 6.5 7.4 7.5 8.0 7.5 6.9 6.8 FD 2 8.2 7.7 MD 2 7.6 6.6 FC 4 FC 4 MC 4 4.3 6.2 FD 2 FC 4 7.0 6.5 7.7 FC 4 6.1 FC 4 8.2 3.9 7.5 FD 2 FC 4 7.1 5.1 FM 3 5.2 7.0 7.9 7.9 6.0 7.2 FD 2 FC 4 6.7 FD 3 FM 4 7.6 7.6 7.5 6.7 7.8 FD 2 B .4 FD 3 FM 4 6.6 6.1 7.8 FM 3 FD 2 FD 2 FM 3 6.0 8.2 FM 3 6.3 RAT/ LITTER # MATERNAL DOSAGE GR OU P II b 1.6 MG/KG/DAY SUBSET D POSTPARTUM DAY 4 POSTCULLING 18946c 18950 18951 18952 18953 18958 18959 18961 18963 18965 18972 18976 18977c 8.7 MC 4 8.8 8.7 8.0 9.3 MC 4 7.8 MC 4 8.2 8.6 8.0 MD 3 6.0 MC 4 8.9 MC 4 8.7 7.8 8.0 8.9 7.0 8.5 MC 4 8.0 MD 2 4.3 7.5 MC 4 8.9 MC 4 10.0 8.0 8.4 6.8 8.4 9.7 6.8 MD 2 MD 4 8.3 MC 4 8.5 8.8 8.9 8.1 8.0 7.4 MC 4 MC 4 MC 4 MD 2 MD 4i MM 3 8.7! MC 4 8.9 9.6 8.3 MC 4 MC 4, 8.4 MC 4 9.5 7.8 MC 4 8.5 FC 4 MC 4 7.3 8.1 8.0 8.7 ; 9.1 7.7 MC 4 MD 2 MD 2 MD 2 8.3 8.8 8.9 9.1 7.8 8.0 9.1 MC 4 7.5 9.1 7.3 MD 2 MD 2 9.1 MC 4 MC 4 MC 4 8.1 MC 4 FC 4 MC 4 8.2 8.3 MD 2 FD 2 MD 2 FC 4 8.9 FC 4 8.8 . 7 MC 4 FC 4 8.0 FC 4 FC 4 7.6 FD 2 5.2 7.2 8.3 FC 4 FC 4 FC 4 8.0 8.5 MC 4 FC 4 7.4 6.7 FD 2 FM 3 6.7 FC 4 7.8 8.0 8.8 FC 4 FC 4 MC 4 FC 4 8.1 FC 4 FD 2 FD 2 8.8 7.5 FC 4 9.0 FC 4 7.3 FC 4 6.5 FC 4 FC 4 FC 4 FD 1 FD 2 FC 4 8.7 7.6 FC 4 8.5 8.4 8.1 FC 4 7.2 FC 4 FC 4 FD 2 7.4 8.6 7.7 7.8 FC 4 7.8 8.3 FC 4 FC 4 FC 4 FC 4 FD 2 FC 4 7.7 FC 4 7.0 8.3 6.9 7.6 8.6 7.4 FD 2 FC 4 7.4 8.9 FM 3 FC 4 6.8 8.0 9.0 FC 4 FD 2 7.8 FC 4 6.1 FC 4 7.0 FC 4 6.6 7.2 FC 4 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 rag/kg/day dosage grou p 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. 20 418-014:PAGE B-88 000373 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 - FI GENERATION PUPS RATS ASSIGNED TO CROSS-FOSTERING PUP # 1 2 3 4 S 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 RAT/ LITTER # MATERNAL DOSAGE GROUP I a 0 (VEHICLE) MG/KG/DAY SUBSET A POSTPARTUM DAY 7 18903 18907 18908 18909 18918b 18920 18924 18929 18934 18936b 18937 18941 18942 MC 4 14.9 12.8 14.0 16.7 12.6 MC 4 MC 4 14.5 13.5 12.2 10.3 MC 4 10.9 11.2 12.2 13.0 MC 4 MC 4 13.6 MC 4 11.9 13.1 12.4 12.5 12.5 11.4 13.9 12.7 MC 4 17.1 11.5 12.6 14.2 MC 4 MC 4 12.2 12.7 12.7 10.5 13.8 MM 4 13.0 15.6 12.3 11.0 13.9 13.2 13.0 11.0 14.6 MC 4 9.5 11.4 10.9 13.2 MC 4 12.5 MC 4 13.3 13.9 12.9 11.7 14.1 MC 4 10.0 13.4 10.6 MC 4 16.7 MC 4 MC 4 14.6 11.5 12.8 11.6 MM 3 MC 4 MM 3 12.0 FC 4 13.6 16.5 12.4 13.0 12.5 MC 4 13.3 FC 4 MM 3 12.2 10 .2 FC 4 12 .4 MD 4 MM 2 MC 4 MM 3 MM 3 FC 4 12 .7 11,.7 MD 2 11 .5 FC 4 11.8 FC 4 14.5 MM 2 12.0 12.3 10.8 13.6 12.5 11.2 9.1 13.0 8.1 11.5 FC 4 13.7 FC 4 11.4 12.3 FC 4 10.4 11.2 11.1 13.7 12.1 FC 4 14.3 11.7 12.9 15.3 10.4 11.6 FC 4 12.5 11.8 11.8 15.2 FC 4 10.2 FC 4 FC 4 14.2 15.6 11.9 12.0 FC 4 13.5 FM 2 FC 4 FM 5 FC 4 11.5 FD 4 FC 4 FC 4 FC 4 13.1 11.5 12.2 12.1 FC 4 FD 2 12.2 8 .2 12 .7 FC 4 FC 4 FM 3 FC 4 13 .0 FD 2 FC 4 FM 3 12.1 FC 4 13.7 FD 2 12.7 12.5 12.2 11.3 11.8 15.1 FC 4 13.8 13.5 15 .2 FD 4 FC 4 RAT/ LITTER # MATERNAL DOSAGE GROUP I C 0 (VEHICLE) MG/KG/DAY SUBSET B POSTPARTUM DAY 7 18901 18904 18906 18912 18917 18919 18922 18923 18931 18932 18935 18938 MC 4 MC 4 MC 4 MC 4 14.1 MC 4 15.0 15.8 12.5 15.3 13.5 13.2 15.0 MC 4 13.7 11.1 12.3 MC 4 15.8 15.7 14.2 MC 4 12.2 12.8 14.7 16.0 12.6 MC 4 MC 4 14.0 15.0 MC 4 14.1 14.5 MC 4 14.7 15.8 14.1 MC 4 13.5 MC 4 17.7 14.5 16.3 MC 4 14.8 13.8 MC 4 17.4 14.2 MC 4 12.3 14 .1 14.7 17.1 17.6 14.5 14.0 14.1 MC 4 15.3 13.5 13.7 MC 4 13.9 16.1 FC 4 15.1 MC 4 13.9 MC 4 14.5 13.6 13.3 13.9 12.8 14.6 15.7 FC 4 FC 4 13.7 13.5 14.1 MC 4 13.2 FC 4 13.6 MC 4 MC 4 15.4 FC 4 FC 4 14.5 13.9 14.0 14.3 14.2 FC 4 FC 4 MC 4 MC 4 14.7 16.0 15.2 FC 4 14.0 FC 4 13.5 FC 4 11.4 FC 4 12.8 14.0 15.1 16.4 14.5 14.2 FC 4 11.9 MC 4 FC 4 11.6 13.8 MC 4 FC 4 15.4 15.0 FC 4 FC 4 13.7 FC 4 13.4 FC 4 14.2 FC 4 12.1 14.6 15.0 13.7 FC 4 FC 4 FC 4 FC 4 14.2 FC 4 FC 4 12 .9 FC 4 13 .0 FC 4 13 ,.9 FC 4 14 4 FC 4 MC 4 FC 4 12.3 12.6 12.8 13.1 FC 4 12.8 FC 4 FC 4 12.4 13.4 15.5 13.3 13.9 FC 4 FC 4 14.3 15.0 13.6 13.2 14.2 14.9 13.8 12.8 13.3 FM 3 12.4 13.3 FC 4 FC 4 14 .3 FC 4 13.7 FD 4 12.,1 FM 4 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 grou p averages. c. Vehicle control group dams cross-fostered with vehicle control litters. 418-014:PAGE B-89 000374 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 - FI GENERATION PUPS RATS ASSIGNED TO CROSS-FOSTERING PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 n 18 19 RAT/ LITTER # MATERNAL DOSAGE GROUP II a 1.6 MG/KG/DAY SUBSET C POSTPARTUM DAY 7 18943 18947 18948 18957 18960 18964 18967 18968 18969 18970 18973 18974 14.1 12.2 6.0 14.1 9.9 11.0 14 .a 15.4 MC 4 MC 4 MC 4 MD 2 12.0 13.3 9.6 11.5 9.5 9.7 14.4 MD 2 12.2 MC 4 13.1 MD 2 12 .4 MC 4 9. 9 11 .,1 11 .5 MC 4 MC 4 13. 1 MC 4 13 ,.3 MC 4 MD 2 13 .6 MC 4 11 .7 MM 3 MM 4 MC 4 MC 4 14 .2 MC 4 13 .4 MC 4 MD 2 12.6 MC 4 11.7 FC 4 MM 4 9.9 13.4 14.2 MC 4 MC 4 10.8 MD 2 FC 4 12.5 11.5 12.7 8.6 10.6 MC 4 12.2 12.6 12.5 12.0 MD 2 FC 4 11.9 MM 4 11.7 8.6 MM 3 13.4 13.9 MC 4 11.4 MC 4 FM 4 12.9 12.7 MM 3 14.0 9.4 10.4 MC 4 12.0 12.4 MC 4 9.9 FM 3 14.7 FC 4 MD 2 12.2 9.7 FC 4 14.1 13.1 MC 4 12.3 12.0 FD 2 12.7 12.7 MD 2 11.5 10.4 11.5 11.8 14.6 12.6 10.6 10.5 FD 2 12.5 12.7 MD 2 11.6 7.3 FC 4 FC 4 MC 4 11.5 10.7 FD 2 FC 4 11.0 9.7 12.7 FC 4 10.5 FC 4 13.5 6.9 12.7 FD 2 FC 4 11.7 8.2 FM 3 11.2 11.4 12.4 12.3 6.3 10.8 FD 2 FC 4 10.3 FD 3 FM 4 10.3 9.9 10.6 11.8 9.4 FD 2 12.4 FD 3 FM 4 11.3 12.8 11.5 FM 3 FD 2 FD 2 FM 3 9.5 13.3 RAT/ LITTER ft MATERNAL DOSAGE GROUP II b 1.6 MG/KG/DAY SUBSET D POSTPARTUM DAY 7 18946c 18950 18951 18952 18953 18958 18959 18961 18963 18965 18972 18976 18977c 10.1 MC 4 12.8 14.6 11.7 11.0 MC 4 14.2 MC 4 13.0 13.4 12.9 MD 3 14.6 MC 4 12.9 MC 4 12.7 12.8 13.3 13.7 13.6 12.7 MC 4 13.5 MD 2 MD 5 13.7 MC 4 15.3 MC 4 13.6 12.2 13.2 12.5 12.5 13.7 12.2 MD 2 MD 4 11 .8 MC 4 14.5 13.1 13.6 13 .1 12.4 10.3 MC 4 MC 4 MC 4 MD 2 MD 4 12.9 12.8 14.7 MC 4 MC 4 13.1 12.9 MC 4 13.0 15.7 11.8 MD 2 MM 3 MC 4 11.9 MC 4 13.1 13.5 MC 4 FC 4 12.4 12.2 15.5 MC 4 MD 2 MD 2 14 .3 12.7 14.7 12.7 11.3 13.2 12.9 MC 4 12.0 15.7 13.4 MD 2 MD 2 13 .8 MC 4 MC 4 MC 4 12 .4 MC 4 FC 4 MC 4 12 .2 12 .0 MD 2 FD 2 MD 2 FC 4 11 .8 FC 4 12 .2 12 .5 MC 4 FC 4 12 .i FC 4 FC 4 11 .4 FD 2 7.8 12.4 10.0 FC 4 FC 4 FC 4 13 .1 13.2 MC 4 FC 4 15.6 11.4 FD 2 FM 3 14 .2 FC 4 13. 6 11 .8 12 .4 FC 4 FC 4 MC 4 FC 4 15..i FC 4 FD 2 FD 2 11.7 12.2 FC 4 12.5 FC 4 12.3 FC 4 11.7 FC 4 FC 4 FC 4 FD 1 FD 2 FC 4 10.4 13.5 FC 4 12.9 12.4 13.9 FC 4 11.9 FC 4 FC 4 FD 2 11.8 11.7 13.3 11.7 FC 4 13.1 12.7 FC 4 FC 4 FC 4 FC 4 FD 2 FC 4 13.5 FC 4 11.5 13.0 11.8 13.2 14.3 13.7 FD 2 FC 4 12.4 11.5 FM 3 FC 4 12.5 12.0 12.8 FC 4 FD 2 11.2 FC 4 11.7 FC 4 13 .1 FC 4 10.9 12.8 FC 4 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 dam3 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. 20 418-014: PAGE B-90 000375 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 - FI 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 18 19 20 RAT/ LITTER It MATERNAL DOSAGE GROUP I a 0 (VEHICLE) MG/KG/DAY SUBSET A POSTPARTUM DAY 14 18903 18907 18908 18909 18918b 18920 18924 18929 18934 18936b 18937 18941 18942 MC 4 26.4 25.8 27.8 33.2 26.1 MC 4 MC 4 28.7 25.1 27.0 30.8 MC 4 26.5 29.3 27.2 28.2 MC 4 MC 4 23.5 MC 4 24.4 25.5 25.5 31.3 26.4 23.4 30.2 27.5 MC 4 33.9 25.4 27.2 33.0 MC 4 MC 4 24.5 28.2 29.3 23.2 26.4 MM 4 28.1 31.0 26.4 25.5 29.8 28.0 24.8 25.2 27.1 MC 4 26.2 27.2 23.7 28.8 MC 4 24.3 MC 4 29.4 30.1 25.8 22.5 23.7 MC 4 24.4 27.3 26.6 MC 4 33.7 MC 4 MC 4 31.9 26.9 24.5 24.5 MM 3 MC 4 MM 3 30.5 FC 4 27.8 33.4 22.7 25.9 26.8 MC 4 24.8 FC 4 MM 3 26.6 24.8 FC 4 25.3 MD 4 MM 2 MC 4 MM 3 MM 3 FC 4 21.7 23.5 MD 2 29.1 FC 4 26.4 FC 4 29.8 MM 2 24.9 27.4 28.6 29.1 22.5 27.1 33.3 28.5 22.4 25.8 FC 4 26.8 FC 4 26.7 26.0 FC 4 24.8 24.0 25.6 22.5 26.7 FC 4 25.7 28.0 28.1 32.3 24.4 25.6 FC 4 28.0 23.7 25.4 31.7 FC 4 24.9 FC 4 FC 4 29.4 30.3 24.4 25.8 FC 4 22.5 FM 2 FC 4 FM 5 FC 4 20.3 FD 4 FC 4 FC 4 FC 4 22.0 25.1 27.3 27.0 FC 4 FD 2 28.1 23.8 26.1 FC 4 FC 4 FM 3 FC 4 26.3 FD 2 FC 4 FM 3 25.4 FC 4 30.6 FD 2 23.9 29.4 28.8 28.1 25.1 28.7 FC 4 27.8 31.0 31.0 FD 4 FC 4 27.5 RAT/ LITTER # MATERNAL DOSAGE GROUP I c 0 (VEHICLE) MG/KG/DAY SUBSET B POSTPARTUM DAY 14 18901 18904 18906 18912 18917 18919 18922 18923 18931 18932 18935 18938 MC 4 MC 4 MC 4 MC 4 29.5 MC 4 32.4 31.6 31.5 27.1 27.4 28.2 35.0 MC 4 27.9 25.8 29.7 MC 4 29.3 28.5 30.9 MC 4 26.8 30.1 29.9 27.2 29.0 MC 4 MC 4 28.6 31.4 MC 4 29.9 27.2 MC 4 29.5 31.2 28.1 MC 4 30.8 MC 4 31.3 29.9 30.1 MC 4 28.2 27.7 MC 4 30.5 32.1 MC 4 30.0 28.2 30.2 30.9 31.0 29.4 27.4 26.6 MC 4 31.7 28.5 30.4 MC 4 26.7 33.3 FC 4 31.2 MC 4 27.2 MC 4 27.9 28.9 28.6 28.7 30.3 30.3 31.3 FC 4 FC 4 31.5 26.4 22.4 MC 4 27.8 FC 4 27.3 MC 4 MC 4 30.6 FC 4 FC 4 32.3 26.7 25.6 28.5 31.0 FC 4 FC 4 MC 4 MC 4 30.6 29.1 25.7 FC 4 26.6 FC 4 32.0 FC 4 27.1 FC 4 31.9 29.6 30.6 32.6 28.7 29.5 FC 4 26.4 MC 4 FC 4 29.1 27.5 MC 4 FC 4 28.9 32.6 FC 4 FC 4 27.7 FC 4 29.1 FC 4 25.8 FC 4 29.9 30.3 28.8 29.1 FC 4 FC 4 FC 4 FC 4 27.9 FC 4 FC 4 28.3 FC 4 26.6 FC 4 30.5 FC 4 27.1 FC 4 MC 4 FC 4 24.9 28.7 29.9 28.3 FC 4 28.7 FC 4 FC 4 26.1 28.9 31.6 25.7 29.5 FC 4 FC 4 31.8 26.8 30.5 26.7 29.0 31.1 27.5 27.9 27.3 FM 3 31.8 24.5 FC 4 FC 4 28.5 FC 4 26.6 FD 4 23.0 FM 4 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. 418-014:PAGE B-91 000376 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 - FI GENERATION PUPS RATS ASSIGNED TO CROSS-FOSTERING PUP It 12 3 4 5 6 7 8 9 10 11 12 13 14 IS 16 17 18 19 RAT/ LITTER # MATERNAL DOSAGE GROUP II a 1.6 MG/KG/DAY SUBSET C POSTPARTUM DAY 14 18943 18947 18948 18957 18960 18964 18967 18968 18969 18970 18973 18974 29.8 25.5 23.4 23.6 25.2 21.8 30.1 30.4 MC 4 MC 4 MC 4 MD 2 26.8 25.5 26.3 27.0 22.1 21.0 29.9 MD 2 26.4 MC 4 26.3 MD 2 28.7 MC 4 27.5 24.1 21.4 MC 4 MC 4 26.0 MC 4 25.5 MC 4 MD 2 27.2 MC 4 24.0 MM 3 MM 4 MC 4 MC 4 25.6 MC 4 26.1 MC 4 MD 2 29.3 MC 4 26.6 FC 4 MM 4 20.1 29.8 24.6 MC 4 MC 4 22.4 MD 2 FC 4 28.0 14.2 24.1 24.1 23.1 MC 4 27.9 26.5 24.8 23.9 MD 2 FC 4 27.0 MM 4 25.1 19.1 MM 3 28.8 30.1 MC 4 22.8 MC 4 FM 4 27.9 25.9 MM 3 23.9 19.2 20.9 MC 4 27.6 27.6 MC 4 20.0 FM 3 31.4 FC 4 MD 2 26.3 21.2 FC 4 28.8 27.0 MC 4 26.2 23.8 FD 2 27.7 23.4 MD 2 24.6 22.1 21.9 28.8 24.4 26.0 24.4 21.5 FD 2 26.6 25.9 MD 2 25.8 16.4 FC 4 FC 4 MC 4 24.0 24.3 FD 2 FC 4 22.3 22.5 27.7 FC 4 22.9 FC 4 28.3 22.1 23.4 FD 2 FC 4 25.5 19.1 FM 3 21.1 20.7 25.7 24.3 15.1 21.3 FD 2 FC 4 23.8 FD 3 FM 4 23.8 23.3 25.6 14.3 22.7 FD 2 27.1 FD 3 FM 4 22.3 21.6 27.9 FM 3 FD 2 FD 2 FM 3 24.9 24.9 FM 3 25.3 RAT/ LITTER MATERNAL DOSAGE GROU P II b 1.6 MG/KG/DAY SUBSET D POSTPARTUM DAY 14 18946c 18950 18951 18952 18953 18958 18959 18961 18963 18965 18972 18976 18977c 35.4 MC 4 24.6 32.4 25.9 26.3 MC 4 26.4 MC 4 26.4 29.2 27.2 MD 3 25.2 MC 4 23.6 MC 4 22.0 21.5 26.9 26.9 28.7 24.9 MC 4 25.8 MD 2 MD 5 24.0 MC 4 28.7 MC 4 24.2 26.1 26.8 26.5 25.8 29.2 24.7 MD 2 MD 4 28.4 MC 4 31.5 25.8 25.1 27.6 25.6 27.5 MC 4 MC 4 MC 4 MD 2 MD 4 28.6 24.1 29.8 MC 4 MC 4 27.5 25.6 MC 4 25.3 33.8 26.2 MD 2 MM 3 MC 4 24.6 MC 4 25.5 26.6 MC 4 FC 4 24.8 25.5 33.7 MC 4 MD 2 MD 2 25.0 24.3 31.0 27.6 24.9 27.3 26.1 MC 4 24.7 31.2 27.0 MD 2 MD 2 28.4 MC 4 MC 4 MC 4 24.5 MC 4 FC 4 MC 4 24.3 30.7 MD 2 FD 2 MD 2 FC 4 20.6 FC 4 24.5 22.0 MC 4 FC 4 26.7 FC 4 FC 4 24.3 FD 2 19.3 23.0 21.6 FC 4 FC 4 FC 4 26.7 23.7 MC 4 FC 4 28.1 23.4 FD 2 FM 3 24.5 FC 4 28.0 25.5 24.0 FC 4 FC 4 MC 4 FC 4 33.6 FC 4 FD 2 FD 2 25.3 23.4 FC 4 24.5 FC 4 26.0 FC 4 27.1 FC 4 FC 4 FC 4 FD 1 FD 2 FC 4 22.7 27.5 FC 4 24.5 24.9 25.7 FC 4 24.7 FC 4 FC 4 FD 2 25.9 19.9 28.0 25.3 FC 4 25.4 27.3 FC 4 FC 4 FC 4 FC 4 FD 2 FC 4 28.9 FC 4 28.0 25.6 24.8 26.1 30.6 25.1 FD 2 FC 4 28.3 24.3 FM 3 FC 4 25.7 24.6 26.4 FC 4 FD 2 28.3 FC 4 26.1 FC 4 26.6 FC 4 25.7 24.7 FC 4 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 L E T T E R - 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. 20 418-014.PAGE B-92 000377 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 DA TA - FI GENERATION PUPS RATS ASSIGNED TO CROSS-FOSTERING PUP (t 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 I a 0 (VEHICLE) MG/KG/DAY SUBSET A POSTPARTUM DAY 21 18903 18907 18908 18909 18918b 18920 18924 18929 18934 18936b 18937 18941 18942 MC 4 41.8 40.9 43.2 53.9 43.5 MC 4 MC 4 43.5 38.7 42.5 41.7 MC 4 44.8 46.3 42.9 43.9 MC 4 MC 4 42.6 MC 4 41.5 39.8 37.5 40.3 43.0 41.8 45.8 42.4 MC 4 57.2 43.2 39.8 49.2 MC 4 MC 4 40.1 45.4 42.9 40.2 43.3 MM 4 44.1 53.2 41.4 40.4 47.3 38.6 39.5 39.9 47.6 MC 4 39.6: 39.2 43.7 40.5 42.6 42.4 41.8 MC 4 MC 4 56.0 37.9 MC 4 MC 4 MC 4 42.7^ 46.7 44.1 42.5 40.4 41.6 41.7 43.3 47.3 MM 3 MC 4; MC 4 MM 3 42.1 FC 4 40.6 29.7 40.8 39.2 50.1 MC 4 39.2 FC 4 MM 3 40.0 37.3 FC 4 36.6 MD 4 MM 2 MC 4 MM 3 MM 3 FC 4 35.6 41.1 MD 2 41.7 FC 4 46.1 FC 4 41.1 MM 2 40.6 44.0 41.2 43.3 35.1 40.2 48.8 40.7 38.7 42.1 FC 4 38.9 FC 4 42.9 40.9 FC 4 40.8 39.2 39.3 48.0 41.0 FC 4 42.3 39.3 37.6 49.3 41.2 41.3 FC 4 40.9 36.4 41.1 39.4 FC 4 40.1 FC 4 FC 4 43.5 51.5 40.6 41.0 FC 4 38.9 FM 2 FC 4 FM 5 FC 4 39.5 FD 4 FC 4 FC 4 FC 4 37.3 41.7 45.2 37.3 FC 4 FD 2 41.8 35.5 37.3 FC 4 FC 4 FM 3 FC 4 42.2 FD 2 FC 4 FM 3 39.9 FC 4 47.5 FD 2 39.6 43.2 44.5 41.3 41.2 51.2 FC 4 47.0 39.2 50.2 FD 4 FC 4 41.7 RAT/ LITTER # MATERNAL DOSAGE GROUP I p 0 (VEHICLE) MG/KG/DAY SUBSET B POSTPARTUM DAY 21 18901 18904 18906 18912 18917 18919 18922 18923 18931 18932 18935 18938 MC 4 MC 4 MC 4 MC 4 42.3 MC 4 49.2 49.6 50.5 43.4 44.2 45.4 50.2 MC 4 49.6 48.7 43.4 MC 4 48.2 50.6 50.2 MC 4 42.2 44.8 49.9 41.1 53.0 MC 4 MC 4 47.2 51.1 MC 4 45.9 43.8 MC 4 43.2 48.9 44.5 MC 4 44.3 MC 4 45.4 49.7 50.3 MC 4 44.3 44.0 MC 4 54.6. 46.6 45.2. 48.7 MC 4. 50.6 50.9; MC 4 4 7.2: 47.3 51.8 51.1 4 5.3 FC 4 50.2 51.4 49.5, MC 4 44.4 42.8 4 5.2 MC 4 MC 4.- 43.5 44.2 42.6 48.3 48.4 46.4 49.5 FC 4 FC 4 48.2 44.0 36.1 MC 4 47.5 FC 4 48.2 MC 4 MC 4 49.4 FC 4 FC 4 50.9 43.7 41.1 46.6 42.5 FC 4 FC 4 MC 4 MC 4 50.0 50.6 44.7 FC 4 42.2 FC 4 45.9 FC 4 39.8 FC 4 50.9 45.7 49.1 45.3 49.3 46.9 FC 4 41.0 MC 4 FC 4 41.3 48.8 MC 4 FC 4 46.7 50.8 FC 4 FC 4 44.2 FC 4 43.0 FC 4 42.2 FC 4 49.6 48.5 43.9 45.0 FC 4 FC 4 FC 4 FC 4 43.3 FC 4 FC 4 49.8 FC 4 45.8 FC 4 44.9 FC 4 43.2 FC 4 MC 4 FC 4 43.9 47.3 47.1 43.9 FC 4 46.2 FC 4 FC 4 42.9 45.5 48.8 40.2 46.8 FC 4 FC 4 50.2 45.5 48.4 39.9 42.0 48.8 46.8 43.3 44.6 FM 3 44.0 42.8 FC 4 FC 4 48.5 FC 4 44.6 FD 4 38.8 FM 4 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 L E T T E R " 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. 418-014:PAGE B-93 000378 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 - FI 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 18 19 RAT/ LITTER H MATERNAL DOSAGE GROUP II a 1.6 MG/KG/DAY SUBSET C POSTPARTUM DAY 21 18943 18947 18948 18957 18960 18964 18967 18968 18969 18970 18973 18974 42.8 45.4 40.7 42.5 34.1 33.1 45.8 50.7 MC 4 MC 4 MC 4 MD 2 42.3 41.5 36.5 39.1 39.5 32.0 46.5 MD 2 40.9 MC 4 37.2 MD 2 45.8 MC 4 37.9 37.4 34.1 MC 4 MC 4 38.0 MC 4 39.8 MC 4 MD 2 39.8 MC 4 40.7 MM 3 MM 4 MC 4 MC 4 44.1 MC 4 41.4 MC 4 MD 2 44.7 MC 4 39.5 FC 4 MM 4 35.2 47.9 48.1 MC 4 MC 4 42.2 MD 2 FC 4 40.8 25.8 41.4 33.4 35.0 MC 4 45.7 39.2 40.6 33.1 MD 2 FC 4 45.0 MM 4 40.1 35.6 MM 3 47.8 40.5 MC 4 37.0 MC 4 FM 4 41.8 42.7 MM 3 38.0 36.9 34.3 MC 4 43.5 42.4 MC 4 34.1 FM 3 41.6 FC 4 MD 2 38.8 27.0 FC 4 47.7 43.1 MC 4 40.7 37.4 FD 2 42.1 41.9 MD 2 37.7 33.2 35.1 40.7 40.2 40.0 37.0 35.3 FD 2 37.9 38.3 MD 2 43.9 30.4 FC 4 FC 4 MC 4 27.0 38.0 FD 2 FC 4 36.1 36.3 39.0 FC 4 33.7 FC 4 40.2 37.5 37.8 FD 2 FC 4 40.4 30.9 FM 3 30.3 35.7 39.3 37.6 36.1 37.4 FD 2 FC 4 41.7 FD 3 FM 4 35.6 39.2 37.2 27.2 33.2 FD 2 46.4 FD 3 FM 4 34.8 45.1 40.2 FM 3 FD 2 FD 2 FM 3 41.1 38.0 FM 3 38.1 RAT/ LITTER # MATERNAL DOSAGE GROUP II b 1.6 MG/KG/DAY SUBSET D POSTPARTUM DAY 21 18946C 18950 18951 18952 18953 18958 18959 18961 18963 18965 18972 18976 18977c 55.0 MC 4 39.7 49.8 44.8 42.8 MC 4 42.4 MC 4 45.5 43.6 40.3 MD 3 42.5 MC 4 40.3 MC 4 37.3 41.4 42.6 40.8 42.8 44.1 MC 4 42.4 MD 2 MD 5 47.3 MC 4 48.7 MC 4 42.1 42.6 42.5 46.2 43.0 49.7 42.5 MD 2 MD 4 42.8 MC 4 46.7 41.0 34.2 43.6 42.4 43.0 MC 4 MC 4 MC 4 MD 2 MD 4 39.9 38.9 44.3 MC 4 MC 4 43.1 41.9 MC 4 43.6 42.0 40.7 MD 2 MM 3 MC 4 40.4 MC 4 44.0 40.6 MC 4 FC 4 41.6 43.6 46.8 MC 4 MD 2 MD 2 47.7 37.9 46.1 40.9 41.5 42.3 41.0 MC 4 43.6 48.5 42.6 MD 2 MD 2 44.6 MC 4 MC 4 MC 4 39.0 MC 4 FC 4 MC 4 41.2 43.4 MD 2 FD 2 MD 2 FC 4 34.5 FC 4 40.4 39.9 MC 4 FC 4 42.4 FC 4 FC 4 39.0 FD 2 36.3 40.3 38.5 FC 4 FC 4 FC 4 39.8 42.6 MC 4 FC 4 44.5 42.3 FD 2 FM 3 37.7 FC 4 43.6 42.9 40.5 FC 4 FC 4 MC 4 FC 4 47.0 FC 4 FD 2 FD 2 37.0 36.3 FC 4 38.6 FC 4 40.5 FC 4 40.7 FC 4 FC 4 FC 4 FD 1 FD 2 FC 4 38.7 43.1 FC 4 36.4 45.6 39.8 FC 4 42.7 FC 4 FC 4 FD 2 44.1 34.6 44.3 39.0 FC 4 43.7 36.8 FC 4 FC 4 FC 4 FC 4 FD 2 FC 4 41.6 FC 4 40.9 40.5 42.8 43.0 42.7 39.9 FD 2 FC 4 44.4 40.7 FM 3 FC 4 41.7 41.0 40.7 FC 4 FD 2 39.6 FC 4 41.9 FC 4 39.0 FC 4 41.5 37.7 FC 4 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 crosa-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. 20 418-014:PAGE B-94 000379 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 - FI GENERATION PUPS RATS ASSIGNED TO PHARAMACOKINETIC EVALUATION PUP ft 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 I 0 (VEHICLE) MG/KG/DAY POSTPARTUM DAY 1 18910 18911 18913 18915 18926 18927 18939 18940 7.3 7.4 7.6 7.2 7.0 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 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 6.0 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 7.2 7.2 7.4 7.4 6.2 6.7 7.2 6.7 6.7 7.2 7.4 6.5 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 6.5 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 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 GROU P II 1.6 MG/KG/DAY POSTPARTUM DAY 1 18956 18971 6.7 6.4 6.2 6.3 6.6 MS MS 7.6 FS 6.1 6.0 6.9 6.3 6.7 4.3 6.1 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 L E T T E R " INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. 418-014: PAGE B-95 000380 PROTOCOL 418-014: ORAL (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 - FI GENERATION PUPS RATS ASSIGNED TO PHARAMACOKINETIC EVALUATION p u p # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 RAT/ LITTER tf MATERNAL DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY POSTPARTUM DAY 4 18910 18911 18913 18915 18926 18927 18939 18940 NO PUP WEIGHTS RECORDED 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 MO 4 7.6 7.6 NO PUP WEIGHTS RECORDED 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 4 NO PUP WEIGHTS RECORDED 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 GR OU P II 1 .6 MG/KG/DAY POSTPARTUM1 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 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. 20 418-014: PAGE B-96 000381 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T - 6295.13) TABLE 30 (PAGE I S ) : PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM INDIVIDUAL DATA - FI GENERATION PUPS RATS ASSIGNED TO PHARAMACOKINETIC EVALUATION pup n 12 34 5 6 7 8 9 10 11 12 13 14 15 16 17 18 RAT/ LITTER It MATERNAL DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY POSTPARTUM DAY 7 18910 18911 18913 18915 18926 18927 18939 18940 18.8 13.2 14.1 13.6 16.4 12.3 12.2 10.3 16.5 12.0 12.6 12.4 15.9 12.0 13.5 12.2 17.1 13.7 9.7 11.3 16.1 12.2 14.1 11.8 16.0 9.9 10.0 13.3 16.6 12.7 12.9 11.1 16.9 11.2 9.3 12.4 15.4 11.7 13.2 8.5 9.9 11.1 13.5 14.9 12.9 12.7 12.1 11.7 9.8 10.6 15.4 11.9 12.0 10.8 13.7 11.7 9.2 14.8 12.4 12.4 10.0 12.7 12.1 14.2 15.9 10.7 11.9 10.3 13.8 11.5 14.9 16.2 10.8 12.9 11.0 14.5 13.2 13.1 15.1 12.4 13.3 12.0 13.0 9.7 14.1 16.5 9.4 12.2 10.3 12.7 10.4 13.4 10.2 13.1 11.6 12.0 7.3 14.5 11.0 13.3 12.0 12.3 MD 6 12.6 12.4 11.1 10.8 FM 7 MD 4 FS 9.1 7.9 10.2 FS FM 4 11.1 RAT/ LITTER MATERNAL DOSAGE GROUP II 1.6 MG/KG/DAY POSTPARTUM DAY 7 18956 18971 13.4 13.2 13.0 13.2 12.7 MS 12.9 12.4 12.4 13.3 12.1 11.8 9.7 MS 9.2 FS 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. 19 20 418-014: PAGE B-97 000382 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 20 (PAGE 16): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM INDIVIDUAL DATA - FI GENERATION PUPS RATS ASSIGNED TO PHARAMACOKINETIC EVALUATION PUP # 1 2 3 4 5 6 7 8 9 10 il 12 13 14 15 16 17 18 RAT/ LITTER # MATERNAL DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY POSTPARTUM DAY 14 18910 18911 18913 18915 18926 18927 18939 18940 35.7 26.7 19.8 24.6 29.7 22.6 24.9 18.3 32.7 25.3 24.2 23.5 31.3 23.2 23.1 16.0 32.9 24.0 21.9 25.2 31.3 23.8 23.7 22.3 33.7 22.1 20.5 24.6 29.4 21.7 21.6 24.5 30.5 17.8 20.2 25.9 28.8 22.5 26.0 16.6 23.0 19.9 24.8 29.5 20.8 23.2 20.2 26.8 23.3 20.5 29.9 23.7 23.1 17.4 19.2 23.0 27.8 29.2 21.3 22.1 18.9 24.2 25.1 26.9 27.8 21.2 25.0 23.7 25.3 20.8 20.7 32.0 22.6 24.1 20.3 26.4 24.1 27.8 30.0 20.6 23.0 13.1 22.7 20.4 25.2 27.8 19.5 21.4 20.6 25.0 18.7 25.3 22.7 24.8 20.0 25.2 14.9 27.8 18.0 21.5 19.2 23.4 FM 7 MD 6 MD 4 25.2 FS 20.0 18.8 19.0 FM10 20.9 21.7 FS FM 4 17.6 RAT/ LITTER # MATERNAL DOSAGE GROUP II 1 .6 MG/KG/DAY POSTPARTUM DAY 14 18956 18971 24.1 23.7 24.4 24.1 24.9 MS 23.7 22.9 24.9 23.7 24.3 23.6 20.7 MS 11.7 FS 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. 19 20 418-014: PAGE B-98 000383 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 FI GENERATION PUPS RATS ASSIGNED TO CROSS -FOSTERING PUP # RAT/ LITTER # 1234 5 6 7 MATERNAL DOSAGE GROUP I a B 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 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 4 F A FC 4 F A F A F A FM 3 18907 M A M A M A M A M A F A F A FC 4 F A F A F A FC 4 FD 4 18908 M A M A M A MM 4 F A F A FC 4 F A FC 4 FC 4 F A FC 4 FC 4 F A F A F A 18909 M A M A MC 4 M A M A MC 4 M A MD 4 F A F A F A F A FC 4 FC 4 FC 4 F A 18918b M A MC 4 M A M A MC 4 M A M A MM 2 MM 2 FC 4 F A F A FC 4 FC 4 F A F A F A 18920 M A MC 4 M A M A M A MC 4 M A MC 4 F A F A F A F A F A FM 3 FD 2 18924 MC 4 M A M A M A MC 4 MC 4 M A MM 3 M A F A F A F A F A FC 4 F A FC 4 FD 4 18929 MC 4 MC 4 M A M A M A M A M A MM 3 F A FC 4 FC 4 FC 4 F A F A F A F A 18934 M A M A MC 4 M A M A M A MC 4 FC 4 F A F A F A F A F A FD 2 18936b M A M A MC 4 M A M A M A M A F A F A F A F A FM 2 18937 M A M A M A M A M A F A FC 4 F A F A F A F A FC 4 FC 4 18941 M A M A M A M A M A MM 3 MM 3 MD 2 F A F A F A FM 5 FD 2 18942 MC 4 M A M A MC 4 MC 4 MC 4 M A M A M A F A FC 4 FC 4 F A FC 4 F A F A FC 4 FA RAT/ LITTER # MATERNAL DOSAGE GROUP :[ c 0 (VEHICLE) MG/KG/DAY SUBSET B 18901 18904 18906 18912 18917 18919 18922 18923 18931 18932 18935 18938 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 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 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 FC 4 FD 4 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 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 MC 4 MC 4 M A M A M A M A M A F A F A F A F A F A M A M A M A M A M A FC 4 FC 4 FC 4 F A F A F A F A FC 4 FC 4 F A 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 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 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 M A M A MC 4 M A M A MC 4 M A F A FC 4 F A FC 4 FC 4 FC 4 F A F A F A 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-fostered with vehicle control litters. 418-014: PAGE B-99 000384 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 31 (PAGE 2): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS RATS ASSIGNED TO CROSS-FOSTERING PUP tt 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 RAT/ LITTER M MATERNAL DOSAGE GROUP II a 1.6 MG/KG/DAY SUBSET C 18943 18947 18948 18957 18960 18964 18967 18968 18969 18970 18973 18974 M A M A M A M A M A FC 4 FC 4 F A F A F A F A FC 4 FC 4 FC 4 F A 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 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 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 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 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 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 M A MD 2 F A F A F A F A F A F A F A F A 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 MC 4 MC 4 M A M A MC 4 M A M A MC 4 M A F A F A F A F A F A FM 3 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 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 F 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 18953 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 F 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 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 1 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 grou p averages. 418-014: PAGE B-100 000385 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 - FI GENERATION PUPS 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 21 22 23 RAT/ LITTER g MATERNAL DOSAGE GROUP I 0 (VEHICLE) MG/KG/DAY 18910 18911 18913 18915 18926 18927 18939 18940 MA M A M A FA FA 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 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 MA M A MA M A FA FA FA FA FA FA FA FA FA FA FA FS FS M A MA M A M A M A MA FA FA FA F A FA FA M A M A M A M A M A M A M A M A F A F A F A F A F A F A F A FM10 FM 4 MA MA MA MA MA M A MA FA FA FA FA FA FA FA FA MA M A MA MA MA MA M A MA MA MA FA FA FA FA FA FA RAT/ LITTER # MATERNAL DOSAGE GROUP II 1.6 MG/KG/DAY 18956 18971 MA MA MA MA MA MS FA FA FA FA FA FA FA MS FA FS 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. 418-014: PAGE B-101 000386 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 - FI GENERATION PUPS RATS ASSIGNED TO CROSS-FOSTERING SUBSET MATERNAL DOSAGE GROUP MATERNAL DOSAGE (MG/KG/DAY) FOSTER LITTER D DAY(S) POSTPARTUM OBSERVATIONS a A Ib 0 (VEHICLE) 18920 3 1/14 PUPS: N O T NURSING. 18941 4 1/9 PUPS: NOT NURSING. B Ic 0 (VEHICLE) 18932 7-10 11-21 1/10 PUPS: 1/10 PUPS: TIP OF TAIL, BLACK. TIP OF TAIL, BLACK. TAIL, BENT (DID NOT EXCEED 2.5 CM FROM BASE OF TAIL).d C II e 1.6 18968 1- 2 3-21 1/10 PUPS: RIGHT HINDPAW, SECOND DIGIT MISSING. 1/9 PUPS: RIGHT HINDPAW, SECOND DIGIT MISSING.d 1B974 2 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 COLOR. 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, . 1.6 mg/kg/day dosage group dams cross -fostered with vehicle control group litters. 418-014:PAGE B-102 000387 PROTOCOL 418-014: ORAL (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 - FI GENERATION PUPS RATS ASSIGNED TO PHARMACOKINETIC EVALUATION MATERNAL DOSAGE GROUP MATERNAL DOSAGE (MG/KG/DAY) LITTER NUMBER D A Y (S ) POSTPARTUM OBSERVATIONS a I 0 (VEHICLE) 18911 14 1/15 PUPS; MOUTH, MASS (0.2 CM X 0.2 CM X 0.2 CM) 18913 G 1/17 PUPS: NOT NURSING. a. Tabulation restricted to adverse observations; all other pups appeared normal. 418-014: PAGE B-103 000388 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 - FI GENERATION PUPS PUPS DYING PRIOR TO CROSS FOSTERING MATERNAL DOSAGE GROUP MATERNAL DOSAGE (MG/KG/DAY) LITTER NUMBER D A Y (S ) POSTPARTUM OBSERVATIONS a I (VEHICLE) 18904 1 3 PUPS: STILLBORN. 18912 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL 18923 1 1 PUP: FOUND DEAD. 18937 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL II 1.6 18952 1 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL 18973 1 1 PUP: FOUND DEAD. NO MILK IN S T O M A C H . BACK: RAISED AREA (2.0 CM X 1.3 CM X 0.3 CM), CONTAINED GAS. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 32) for external observations confirmed at necropsy. 418-014: PAGE B-104 000389 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 - FI GENERATION PUPS RATS ASSIGNED TO CROSS -FOSTERING SUBSET MATERNAL DOSAGE GROUP MATERNAL DOSAGE (MG/KG/DAY) FOSTER LITTER # D A Y (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 STOMACH. TISSUES APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL OTHER 18908 4 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 18909 18918 4 21 ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 18920 2 4 1 PUP: FOUND DEAD. NO MILK IN STOMACH. TISSUES APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL OTHER 18924 4 21 1 PUP: FOUND DEAD. NO MILK IN STOMACH. TISSUES APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL OTHER 18934 2 21 1 PUP: FOUND DEAD. NO MILK IN STOMACH. TISSUES APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL OTHER 18936 21 ALL PUPS APPEARED NORMAL. 18941 2 2 PUPS: FOUND DEAD. ALL TISSUES APPEARED NORMAL. 18942 4 ALL PUPS APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the 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. 418-014: PAGE B-105 000390 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 - FI GENERATION PUPS RATS ASSIGNED TO CROSS-FOSTERING SUBSET MATERNAL DOSAGE GROUP MATERNAL DOSAGE (MG/KG/DAY) FOSTER LITTER # D A Y (S ) POSTPARTUM OBSERVATIONS a B Ib 0 (VEHICLE) 18901 18904 4,21 4 ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 18906 4 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 18917 18922 18923 18931 18932 18935 21 21 21 21 4 21 ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the individual pup clinical observations table (Table 32) for external observations confirmed at necropsy. b. Vehicle control group dams cross-fostered vehicle control group litters. 418-014: PAGE B-106 000391 PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) T A B L E 33 (PAGE 4): N E C R O P S Y O B S E R V A T I O N S - I N D I V I D U A L D A T A FI G E N E R A T I O N PUPS RATS ASSIGNED TO CROSS -FOSTERING SUBSET MATERNAL DOSAGE GROUP MATERNAL DOSAGE (MG/KG/DAY) FOSTER LITTER # DAY (S ) POSTPARTUM OBSERVATIONS a C II b 18943 18947 21 4 ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 18948 2 3 3 PUPS: 2 PUPS: FOUND DEAD. NO MILK IN STOMACH. A L L OTHER TISSUES APPEARED NORMAL. FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. 19857 4 ,21 ALL PUPS APPEARED NORMAL. 19860 19864 21 4,21 ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 19868 2 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 19870 21 ALL PUPS APPEARED NORMAL. 18973 2 4,21 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL PUPS APPEARED NORMAL. 18974 2 13 PUPS: FOUND DEAD. NO MILK IN STOMACH OTHER TISSUES APPEARED NORMAL. ALL a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the 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. 418-014:PAGE B-107 000392 PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 33 (PAGE 5): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS RATS ASSIGNED TO CROSS-FOSTERING SUBSET MATERNAL DOSAGE GROUP MATERNAL DOSAGE (MG/KG/DAY) FOSTER LITTER # DAY(S) POSTPARTUM OBSERVATIONS a D II b 1.6 18946 2 5 2 PUPS: 7 PUPS: 1 PUP: FOUND DEAD. ALL TISSUES APPEARED NORMAL. FOUND DEAD. NO MILK IN STOMACH. A L L OTHER TISSUES APPEARED NORMAL. :FOUND DEAD. ALL TISSUES APPEARED NORMAL. 18951 18952 18953 18958 18961 18963 18965 18972 21 21 4,21 4,21 4,21 4 4 21 ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 18976 2 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 18977 2 3 11 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. 1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL. a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. Refer to the 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. 418-014:PAGE B-108 000393 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 - FI GENERATION PUPS RATS ASSIGNED TO PHARMACOKINETIC EVALUATION MATERNAL DOSAGE GROUP MATERNAL DOSAGE (MG/KG/DAY) LITTER NUMBER D A Y (S ) POSTPARTUM OBSERVATIONS a I 0 (VEHICLE) 18910 18911 14 14 ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 18913 4 14 1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION. ALL PUPS APPEARED NORMAL. 18915 1 14 2 PUPS: STILLBORN. ALL PUPS APPEARED NORMAL. 18926 18927 18939 18940 14 14 14 14 ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. II 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. Refer to the individual pup clinical observations table (Table 32) for external observations confirmed at necropsy. 418-014: PAGE B-109 000394 APPENDIX C PROTOCOL AND AMENDMENT 000395 418-014: PAGE C-1 O P r im e d ic a ArgusResearchLaboratories, Inc. 905SheehyDrive, BuildingA Horsham, PA19044 Telephone:(215) 443-8710 Telefax:(215) 443-8587 PROTOCOL 418-014 SPONSOR'S STUDY NUMBER: T-6295.13 STUDY TITLE: Oral (Gavage) Cross-Fostering Study of PFOS in Rats PURPOSE: The purpose of this study is to evaluate pup survival of F1 rats following PFOS treatment of Cri:CDBR VAF/Plus female rats during premating, gestation and lactation. F1 pups will be cross-fostered during the lactation period to differentiate via maternal milk the differences for in utero exposure and exposure through maternal milk. Selected pharmacokinetic samples will be collected from Fo generation females and F1 generation pups. TESTING FACILITY: Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, Pennsylvania 19044-1297 Telephone: (215)443-8710 Telefax: (215)443-8587 STUDY DIRECTOR: Raymond G. York, Ph.D., DABT Associate Director of Research SPONSOR: 3M Corporate Toxicology 3M Center, Building 220-2E-02 St. Paul, Minnesota 55144-1000 STUDY MONITOR: Marvin T. 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 000396 418-014: PAGE C-2 Protocol418-014 Page2 REGULATORY CITATIONS: U.S. Food and Drug Administration (1994). International Conference on Harmonisation; Guideline on detection of toxicity to reproduction for medicinal products. Federal Register, September 22, 1994, Vol. 59, No. 183. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. Japanese Ministry of Health and Welfare (1997). Good Laboratory Practice Standard for Safety Studies on Drugs, 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 decision/recommendation on compliance with principles of good laboratory practice. Official Journal of the European Communities: Legislation. 32 (No. L 315; 28 October): 1-17. REGULATORY COMPLIANCE: This study will be conducted in compliance with the Good Laboratory Practice (GLP) regulations cited above. All changes or revisions of this protocol shall be documented, signed by the Study Director and the Sponsor, dated and maintained with the protocol. The Quality Assurance Unit (QAU) will audit the protocol, the raw data and the report, and will inspect critical phases of the study in accordance with the Standard Operating Procedures of Argus Research Laboratories, Inc. The final report will include a statement signed by the Study Director that the report accurately reflects the raw data obtained during the performance of the study and that all applicable GLP regulations were followed in the conduct of the study. Should significant deviations from GLP regulations occur, each will be described in detail, together with how the deviation might affect the quality or integrity of the study. SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE: See ATTACHMENT 1 to the protocol. 000397 418-014: PAGE C-3 Protocol418-014 Page3 TEST ARTICLE AND VEHICLE. Identification: Test Article: Name: Physical Description: Lot/Batch Number Specific Gravity: Purity: Expiration Date: PFOS (Synonym: FC-95). Light-colored powder. 217. - 0.6. 98.9%. May 2000. Information on the identity, composition, strength and purity of the test article is on file with the Sponsor. Vehicle: 0.5% Tween 80 in Reverse Osmosis Membrane Processed Deionized Water (R.O. Deionized Water). Supplier and lot identification of Tween 80 to be documented in the raw data. Neither the Sponsor nor the Study Director is aware of any potential contaminants likely to be present in the vehicle components that would interfere with the results of this study. Therefore, no analyses other than those mentioned in this protocol will be conducted. Safety Precautions: Gloves, mask, appropriate eye protection and a uniform/lab coat are to be worn during formulation preparation and administration. The Material Safety Data Sheet (MSDS) is attached to the protocol (ATTACHMENT 2). Storage: Bulk Test Article: Vehicle Components: Prepared Vehicle: Prepared Formulations: Room temperature. Room temperature. Room temperature. Room temperature. 000398 418-014: PAGE C-4 Protocol418-014 Page4 All test article shipments to the Testing Facility should be addressed to the attention of Julian Gulbinski, Manager of Formulations, at the previously cited address and telephone number. Shipments should include information concerning storage conditions and shipping cartons should be labeled appropriately. The recipient should be notified in advance of shipment. FORMULATION: Frequency of Preparation: Formulations (suspensions) will be prepared daily at the Testing Facility. Data verifying the stability of the test article in the vehicle for 48 hours under the conditions of administration are on file with the Sponsor. Detailed preparation procedures are attached to this protocol (ATTACHMENT 3). Adjustment for Purity: The test article will be considered 100% pure for the purpose of dosage calculations. Testing Facility Reserve Samples: The Sponsor will reserve a sample (1 g) of each lot of the bulk test article used during the course of this study. The Testing Facility will reserve a sample (5 mL) of each lot of the vehicle components used during the course of this study. Samples will be stored under the previously cited conditions. ANALYSES: Samples additional to those described below may be taken if deemed necessary during the course of the study. Bulk Test Article Sampling: No analyses of the bulk test article will be conducted during the course of this study. Information on the stability of the bulk test article is on file with the Sponsor. Analyses of Prepared Formulations: Homogeneity and stability of prepared formulations is on file with the Sponsor. However, records will be maintained to document how the test article formulations were prepared. 000399 418-014: PAGE C-5 Protocol418-014 Page5 Concentration of Test Article Formulations: Concentration of the prepared formulations will be verified during the course of this study. Duplicate samples (2 mL each) will be taken from the first and last preparation on the day prepared. One sample of each set will be shipped for analysis; the remaining samples will be retained at the Testing Facility as backup samples. Backup samples will be stored frozen (-70C or below) and discarded at the Testing Facility upon request of the Sponsor. Shipping Instructions: Samples to be analyzed will be shipped (frozen on dry ice) to: Kris J. Hansen, Ph.D. 3M Environmental Technology and Safety Services 935 Bush Avenue Building 2-3E-09 St. Paul, Minnesota 55133-3331 Telephone; (612) 778-6018 Telefax: (612)778-6176 The recipient will be notified in advance of sample shipment. DISPOSITION: Prepared formulations will be discarded at the Testing Facility. All remaining bulk test article will be returned to the Study Monitor at the previously cited address upon completion of all work with the test article. TEST SYSTEM: Species/Strain and Reason for Selection: The Crl:CDBR VAF/Plus (Sprague-Dawley) rat was selected as the Test System because: 1) this strain of rat was used in the reproductive and developmental toxicity studies; 2) historical data and experience exist at the Testing Facility*1'35; and 3) the test article is pharmacologically active in the species and strain. 000400 418-014: PAGE C-6 Protocol418-014 Page6 Number: Initial population acclimated: Population selected for study: Population selected for pharmacokinetic sample collection (day 14 postpartum): 86 virgin female rats. 54 mated female rats (30 in the control group and 24 in the treated group). 12 mated female rats (six per dosage group). Body Weight and Age: Female rats will be ordered to have body weights of 200 g to 225 g each at receipt, at which time they will be expected to be at least 60 days of age. Actual body weights will be recorded the day after receipt and will be documented in the raw data. The weight range will be included in the final report. Sex: Female rats will be given the test article. Male rats of the same source and strain will be used only as breeders and are not considered part of the Test System. Source: Charles River Laboratories, Inc. The rats will be shipped in filtered cartons by air freight and/or truck from Charles River Laboratories, Inc., to the Testing Facility. Identification: Fo Generation: Rats are permanently identified using Monel self-piercing ear tags (Gey Band and Tag Co., Inc., No. MSPT 20101). Male rats are given unique permanent identification numbers upon assignment to the Testing Facility's breeder male rat population. Female rats are assigned temporary numbers at receipt and given unique permanent identification numbers when assigned to the study. F1 Generation: Pups will not be individually identified during lactation; all parameters will be evaluated in terms of the litter. 000401 418-014: PAGE C-7 Protocol418-014 Page7 ANIMAL HUSBANDRY: All cage sizes and housing conditions are in compliance with the Guide for the Care and Use of Laboratory Animals{4). Housing: Fo Generation Rats/F1 Generation Litters: Fo generation rats will be individually housed in stainless steel, wire-bottomed cages, except during the cohabitation and postpartum periods. During cohabitation, each pair of rats will be housed in the male rafs cage. Beginning no later than day 20 of presumed gestation, Fo generation female rats will be individually housed in nesting boxes, except during collection intervals for urine and fecal samples. During these collection intervals, the female rats will be housed individually in metabolism cages. Each dam and assigned litter will be housed in a common nesting box during the postpartum period. Nesting Material: Bedding material (bed-o'cobs) will be provided. Bedding will be changed as often as necessary to keep the animals dry and clean. Analyses for possible contamination are conducted annually and documented in the raw data. Room Air, Temperature and Humidity: The animal room is independently supplied with at least ten changes per hour of 100% fresh air that has been passed through 99.97% HEPA filters (Airo Clean room). Room temperature will be maintained at 64F (18C) to 79F (26C) and monitored constantly. Room humidity will also be monitored constantly and maintained at 30% to 70%. Light: An automatically controlled 12-hour light: 12-hour dark fluorescent light cycle will be maintained. Each dark period will begin at 1900 hours EST. Diet: Rats will be given Certified Rodent Diet #5002 (PMI Nutrition International) available ad libitum from individual feeders. 000402 418-014: PAGE C-8 Protocol418-014 Page8 Water Water will be available ad libitum from individual bottles attached to the cages or from an automatic watering access system. All water will be from a local source and passed through a reverse osmosis membrane before use. Chlorine will be added to the processed water as a bacteriostat; processed water is expected to contain no more than 1.2 ppm chlorine at the time of analysis. Water is analyzed monthly for possible bacterial contamination and twice annually for possible chemical contamination. Contaminants: Neither the Sponsor nor the Study Director is aware of any potential contaminants likely to be present in the certified diet, the drinking water or the nesting material at levels that would interfere with the results of this study. Therefore, no analyses other than those routinely performed by the feed supplier or those mentioned in this protocol will be conducted. RANDOMIZATION. COHABITATION AND CROSS-FOSTERING PROCEDURES: Upon arrival, male and female rats will be assigned to individual housing on the basis of computer-generated random units. After acclimation, 78 virgin female rats will be selected for study on the basis of physical appearance and body weights recorded during acclimation. The female rats will be assigned to dosage groups (42 in the control group and 36 in the treated group) based on computer-generated (weightordered) randomization procedures and treated with either the vehicle or the test article for 42 days prior to cohabitation. Within each dosage group, consecutive order will be used to assign female rats to cohabitation with breeder male rats, one male rat per female rat. The rats assigned to the control group will be cohabited one day prior to cohabitation for rats assigned to the treated group. The cohabitation period will consist of a maximum of five days. Female rats with spermatozoa observed in a smear of the vaginal contents and/or a copulatory plug observed in situ will be considered to be at day 0 of presumed gestation and assigned to individual housing. Mated female rats (those with confirmed evidence of mating) will be assigned as follows: of the 42 female rats assigned to the control group, 30 mated female rats will be assigned to the control group for cross-fostering purposes; of the 36 female rats assigned to the treated group, 24 mated female rats will be assigned to the treated group for cross-fostering purposes. An additional six mated female rats from each dosage group will be designated for pharmacokinetic sample collection (as described below) following delivery, but these rats will not be cross-fostered. Finally, any remaining female rats with confirmed evidence of mating that were assigned to the control group prior to cohabitation will be assigned to the control pool for cross-fosteoig 418-014.PAGE C-9 Protocol418-014 Page9 purposes. Any remaining female rats with confirmed evidence of mating that were assigned to the treated group prior to cohabitation will be sacrificed and discarded without further evaluation at the discretion of the Study Director and the Study Monitor. Day 1 of lactation (postpartum) is defined as the day of birth and is also the first day on which all pups in a litter are individually weighed (pup body weights will be recorded after all pups in a litter are delivered and groomed by the dam). Twelve dams and their respective litters will be selected and assigned to the pharmacokinetic sample collection (six dams and litters per dosage group). These litters will not be cross-fostered. Up to 24 litters per dosage group will be reassigned for cross-fostering on day 1 postpartum. The litters will be assigned such that four subsets are obtained among the two dosage groups as described in the table below. Dosage Number of Mated Dosage Number of Reassigned Utter Type Subset Group Female Rats (mg/kg/day) Litters Reassigned (Dam Treatment) Number I 12 0 (Vehicle) 12 I 12 0 (Vehicle) 12 II 12 1.6 12 Treated Untreated Treated A B C II 12 1.6 12 Untreated 0 Detailed procedures for reassignment of litters for cross-fostering are attached to this protocol (ATTACHMENT 4). On day 1 postpartum, pups from dams assigned to the control pool will be culled, sacrificed via decapitation and handled as described in ATTACHMENT 4. On day 4 postpartum, cross-fostered litters will be culled to five male and five female pups per litter, where possible. Pups not selected for continued evaluation will be sacrificed via decapitation; the lungs (saved in Bouin's solution) and the liver (saved in neutral buffered 10% formalin) will be collected from the first ten culled pups from each dosage group (irrespective of litter) determined to be at day 4 postpartum and preserved for possible future histopathological evaluation. Remaining culled pups will be sacrificed via decapitation and discarded without necropsy evaluation. 000404 418-014.PAGE C-10 Protocol418-014 Page10 ADMINISTRATION: Route and Reason for Choice: The oral (gavage) route was selected for use because: 1) this was the route of administration in the developmental and reproductive toxicology studies; and 2) it is one of the possible routes of human exposure. Method and Frequency. Fo Generation Female Rats: Female rats will be given the test article or the vehicle once daily beginning 42 days prior to cohabitation through day 21 postpartum. Dosages will be adjusted daily for body weight changes and given at approximately the same time each day. F1 Generation Puds: F1 generation pups will not be directly given the test article, but may be possibly exposed to the test article during maternal gestation (in utero exposure) or via maternal milk during the lactation period. Rationale for Dosage Selection: Dosages were selected on the basis of a previous study conducted with the test article (Argus Research Laboratories, Inc., Protocol 418-008). Dosage Levels. Concentrations and Volumes: Dosage Number of Mated Dosage Group Female Rats (mg/kg/day) I 30 + 6* 0 (Vehicle) II 24 + 6* 1.6 Concentration (mg/mL) 0 0.32 Dosage Volume (mUkg) Argus Batch Number 5 B-418-014-A(Day.Month.Year) 5 B-418-014-8(Day.Month.Year) The test article will be considered 100% pure for (tie purpose of dosage calculations. a. Samples for analysis of pharmacokinetics will be collected from six dams and their respective litters on day 14 postpartum. These dams and their respective litters will be sacrificed following sample collection. TESTS. ANALYSES AND MEASUREMENTS - Fo GENERATION: Viability: All Periods: At least twice daily. 000405 418-014: PAGE C-11 Protocol418-014 Page11 Clinical Observations and/or General Appearance: Acclimation Period: At least once. Dosage Period: Twice daily. Prior to administration and once approximately one hour postdosage. Maternal Behavior Days 1, 4, 7, 14 and 21 postpartum. Observed abnormal behavior will be recorded daily. Clinical observations may be recorded more frequently than cited above, if deemed appropriate by the Study Director and/or Study Monitor. Body Weights: Acclimation Period: At least once. Dosage Period: Daily. Sacrifice: Terminal weight. Feed Consumption Values (recorded and tabulated): Acclimation Period: At least once. Dosage Period: Weekly to cohabitation. Daily during presumed gestation and days 1, 4, 7,10 and 14 postpartum. Feed consumption will not be tabulated after day 14 postpartum, when it is expected that pups will begin to consume maternal feed. Feed consumption values may be recorded more frequently if it is necessary to replenish the feed. These intervals will not be tabulated. Mating Performance: Mating will be evaluated daily during the cohabitation period and confirmed by observation of spermatozoa in a smear of the vaginal contents and/or a copulatory plug observed in situ. 000406 418-014:PAGE C-12 Protocol418-014 Page12 Natural Delivery: Female rats will be evaluated for Clinical Observations During Parturition [to be performed under red light conditions during the dark period (time each pup is observed will be recorded)]. Duration of Gestation (day 0 of presumed gestation to the time the first pup is observed). Length of Parturition (time of observation of last pup minus the time of observation of the first pup divided by N-1 pups in each litter). Litter Size (defined as all pups delivered). Pup Viability at Birth. Pharmacokinetic Sample Collection: Caps and labeled tubes will be weighed (combined weight, to the nearest .001 gram) before and after retention of pooled pup samples for subsequent use in pharmacokinetic analyses. These weights will be documented in the raw data, and copies of these weights will be included with the packing list prior to shipment. Blood. Milk and Liver Samples - Fo Generation Rats: Blood, milk and liver samples will be collected from six maternal rats designated for pharmacokinetic sample collection per dosage group on day 14 postpartum. The time of sample collections (blood and milk) will be recorded in the raw data. Milk samples will be collected approximately two to six hours postdosage on day 14 postpartum. Each dam will be removed from the nesting box and individually housed for approximately four hours. The dam will be injected intravenously with one unit of oxytocin approximately five minutes before milk samples (at least 100 pL per sample) are collected. The samples will be immediately frozen on dry ice and maintained frozen (-70C or below) until shipment to the Sponsor for analysis. Following milk sample collection, blood samples (approximately 4 mL each) will be collected from the maternal rats via the inferior vena cava and transferred into serum separator tubes. The samples will be spun in a refrigerated centrifuge. The serum will be transferred into polypropylene tubes labeled with the study number, animal identification, date of collection, study day and collection timepoint. All samples will be immediately frozen on dry ice and maintained frozen (-70C or below) until shipment to the Sponsor for analysis. 000407 418-014: PAGE C-13 Protocol418-014 Page13 Following collection of milk and blood samples, a liver section (right lateral lobe) and the milk-secreting glands from the axillary, thoracic, abdominal and inguinal regions of each dam (left side only) will be collected, frozen and stored (-70C or below) until shipment to the Sponsor for analysis. Blood and Liver Samples - F1 Generation Puds: Blood samples will be collected via the inferior vena cava from each pup on day 14 postpartum, pooled (per litter) and transferred into serum separator tubes. The samples will be spun in a refrigerated centrifuge. The serum will be transferred into polypropylene tubes labeled with the study number, animal identification, date of collection, study day and collection timepoint. All samples will be immediately frozen on dry ice and maintained frozen (-70C or below) until shipment to the Sponsor for analysis. The liver from each pup will be collected, pooled (per litter), frozen and stored (-70C or below) until shipment to the Sponsor for analysis. Shipping Instructions: All samples will be maintained frozen (-70C or below) until shipment for analysis. Samples will be shipped on frozen on dry ice via overnight mail. A packing list will be included with the samples and sent to Kris J. Hansen, Ph.D., at the previously cited address. Both the recipient and the Study Monitor will be notified in advance of sample shipment. Urine and Fecal Sample Collection: Fo generation female rats will be housed individually in metabolism cages for collection of urine and fecal samples from days 21 to 22 postpartum. Following the 24-hour collection interval, samples will be collected into centrifuge tubes, placed on dry ice and stored frozen (-70 C or below) until shipment for analysis. Shipping Instructions: All samples will be maintained frozen (-70C or below) until shipment for analysis. Samples will be shipped on frozen on dry ice via overnight mail. A packing list will be included with the samples and sent to Kris J. Hansen, Ph.D., at the previously cited address. Both the recipient and the Study Monitor will be notified in advance of sample shipment. METHOD OF SACRIFICE - Fo GENERATION RATS: Rats will be sacrificed by carbon dioxide asphyxiation. 000408 418-014: PAGE C-14 Protocol418-014 Page14 NECROPSY Fo GENERATION RATS: Gross lesions will be retained in neutral buffered 10% formalin for possible future evaluation (a table of random units will be used to select one control group rat from which all tissues examined at necropsy will be retained, in order to provide control tissues for any possible histopathological evaluations of gross lesions). Unless specifically cited below, all other tissues will be discarded. Rats Not Delivering a Litter Rats that do not deliver a litter will be sacrificed on day 25 of presumed gestation and examined for gross lesions. Uteri will be stained with 10% ammonium sulfide to confirm the absence of implantation sites,5). Scheduled Sacrifice - Rats Assigned to Pharmacokinetic Sample Collection: On day 14 postpartum, milk and blood samples, a liver section (right lateral lobe) and the milk-secreting glands from the axillary, thoracic, abdominal and inguinal regions of each dam (left side only) will be collected as described previously, frozen and stored (-70C or below) until shipment to the Sponsor for analysis. Following the completion of pharmacokinetic sample collection, Fo generation female rats will be sacrificed, and a gross necropsy of the thoracic, abdominal and pelvic viscera will be performed. The number and distribution of implantation sites will be recorded. Scheduled Sacrifice - Day 22 Postpartum: On day 22 postpartum, blood samples will be collected from the Fo generation female rats following removal from metabolism caging. Blood samples (approximately 4 mL each) will be collected from the maternal rats via the inferior vena cava and transferred into serum separator tubes. The samples will be spun in a refrigerated centrifuge. The serum will be transferred into polypropylene tubes labeled with the study number, animal identification, date of collection, study day and collection timepoint. All samples will be immediately frozen on dry ice and maintained frozen (-70C or below) until shipment to the Sponsor for analysis. The female rats will then be sacrificed, and a gross necropsy of the thoracic, abdominal and pelvic viscera will be performed. The number and distribution of implantation sites will be recorded. A liver section (right lateral lobe) from each dam will be collected, frozen and stored (-70 C or below) until shipment to the Sponsor for analysis. 000409 418-014: PAGE C-15 Protocol418-014 Page15 Dams with No Surviving Puds: Dams with no surviving pups will be sacrificed after the last pup is found dead, missing or presumed cannibalized. Prior to sacrifice, a blood sample (approximately 4 mL) will be collected from the maternal rat via the inferior vena cava and transferred into serum separator tubes. The sample will be spun in a refrigerated centrifuge. The serum will be transferred into a polypropylene tube labeled with the study number, animal identification, date of collection, study day and collection timepoint. The sample will be immediately frozen on dry ice and maintained frozen (-70C or below) until shipment to the Sponsor for analysis. A gross necropsy of the thoracic, abdominal and pelvic viscera will be performed. A liver section (right lateral lobe) from each dam will be collected, frozen and stored (-70C or below) until shipment to the Sponsor for analysis. Postpartum data for these dams will be excluded from summary tables. Rats Found Dead or Moribund: Rats that die or are sacrificed because of moribund condition, abortion or premature delivery will be examined for the cause of death or moribund condition on the day the observation is made. The rats will be examined for gross lesions. A liver section (right lateral lobe) from each dam will be collected, frozen and stored (-70C or below) until shipment to the Sponsor for analysis. Pregnancy status and uterine contents of female rats will be recorded. Aborted fetuses and/or delivered pups will be examined to the extent possible. Uteri of apparently nonpregnant rats will be stained with 10% ammonium sulfide to confirm the absence of implantation sites<5). Shipping Instructions: All samples will be maintained frozen (-70C or below) until shipment for analysis. Samples will be shipped frozen on dry ice via overnight mail. A packing list will be included with the samples and sent to Kris J. Hansen, Ph.D., at the previously cited address. Both the recipient and the Study Monitor will be notified in advance of sample shipment. TESTS. ANALYSES AND MEASUREMENTS - F1 GENERATION: Viability: Postpartum Period: Litters will be observed for dead pups at least twice daily. The pups in each litter will be counted once daily' 000410 418-014.PAGE C-16 Protocol418-014 Page16 Clinical Observations and/or General Appearance: Postpartum Period: Once daily. Clinical observations may be recorded more frequently than cited above, if deemed appropriate by the Study Director and/or the Study Monitor. Body Weights: Postpartum Period: Days 1 (birth), 4, 7, 14 and 21 postpartum. Sacrifice: Terminal weight. METHOD OF SACRIFICE - F1 GENERATION RATS: As previously cited for Fo generation rats. Culled pups will be sacrificed via decapitation. NECROPSY - F1 GENERATION RATS: Gross lesions will be retained in neutral buffered 10% formalin for possible future evaluation (a table of random units will be used to select one control group rat of each sex from which all tissues examined at necropsy will be retained, in order to provide control tissues for any possible histopathological evaluations of gross lesions). Unless specifically cited below, all other tissues will be discarded. As described previously, caps and labeled tubes will be weighed (combined weights, to the nearest .001 gram) before and after retention of pooled pup samples. These weights will be documented in the raw data, and copies of these weights will be included with the packing list prior to shipment. Puds Found Dead on Dav 1 Postpartum: Pups that die before examination of the litter for pup viability will be evaluated for vital status at birth. The lungs will be removed and immersed in water. Pups with lungs that sink will be identified as stillborn; pups with lungs that float will be identified as livebom, and to have died shortly after birth. Pups with gross lesions will be preserved in Bouin's solution for possible future evaluation. The lungs will be preserved in Bouin's solution and the liver will be preserved in neutral buffered 10% formalin for possible future evaluation. 000411 418-014: PAGE C-17 Protocol418-014 Page17 Pups Found Dead or Moribund on Days 2 to 21 Postpartum: Pups found dead or sacrificed because of moribundity will be examined for gross lesions and for the cause of death or the moribund condition. Pups with gross lesions found on days 2 to 4 postpartum will be preserved in Bouin's solution for possible future evaluation; gross lesions of pups found on days 5 to 21 postpartum will be preserved in neutral buffered 10% formalin. For all pups found dead on days 2 to 4 postpartum, the lungs will be preserved in Bouin's solution and the liver will be preserved in neutral buffered 10% formalin for possible future evaluation. For all pups found dead on days 5 to 21 postpartum, the lungs and the liver will be preserved in neutral buffered 10% formalin for possible future evaluation. Pups Not Selected for Continued Observation - Davs 1 and 4 Postpartum: All pups culled on day 1 postpartum (pups from dams assigned to the control pool) and day 4 postpartum (all other culled pups) will be sacrificed via decapitation. The lungs and the liver will be collected from the first ten pups culled (irrespective of litter) from each dosage group on each day (control pool only on day 1 postpartum); the lungs will be retained in Bouin's solution, and the livers will be retained in neutral buffered 10% formalin for possible future evaluation. Remaining culled pups will be sacrificed and discarded without evaluation. Scheduled Sacrifice - Litters Assigned to Pharmacokinetic Sample Collection: On day 14 postpartum, litters assigned to the pharmacokinetic sample collection will be sacrificed with their respective dams. Individual, pooled samples (per litter) of blood and liver will collected as described previously. The pups will be examined for gross lesions. Scheduled Sacrifice - Litters Assigned to Sacrifice on Day 21 Postpartum: On day 21 postpartum, six litters per subset will be randomly selected for collection of pooled samples (per litter) of blood and liver. Blood samples will be collected via the inferior vena cava from each pup, pooled (per litter) and transferred into serum separator tubes. The samples will be spun in a refrigerated centrifuge. The serum will be transferred into polypropylene tubes labeled with the study number, animal identification, date of collection, study day and collection timepoint. All samples will be immediately frozen on dry ice and maintained frozen (-70C or below) until shipment to the Sponsor for analysis. 000412 418-014: PAGE C-18 Protocol418-014 Page18 The pups will be examined for gross lesions. The liver from each pup will be collected, pooled (per litter), frozen and stored (-70C or below) until shipment to the Sponsor for analysis. All other remaining pups will be sacrificed and discarded without further evaluation. Shipping Instructions: All samples will be maintained frozen (-70C or below) until shipment for analysis. Samples will be shipped frozen on dry ice via overnight mail. A packing list will be included with the samples and sent to Kris J. Hansen, Ph.D., at the previously cited address. Both the recipient and the Study Monitor will be notified in advance of sample shipment. STATISTICAL EVALUATION: Averages and percentages will be calculated. Litter values will be used where appropriate. Additional procedures and/or analyses may be performed, if deemed appropriate. DATA ACQUISITION. VERIFICATION AND STORAGE: Data will be hand- and/or computer-recorded. Records will be reviewed by the Study Director and/or appropriate management personnel within 21 days after generation. All original records will be stored in the archives of the Testing Facility. All original data will be bound and indexed. A copy of all raw data will be supplied to the Sponsor upon request. Preserved tissues will be stored at the Testing Facility at no charge for one year after mailing of the draft final report, after which time the Sponsor will be contacted to determine the disposition of these materials. 000413 418-014: PAGE C-19 Protocol418-014 Page19 RECORDS TO BE MAINTAINED: Protocol and Amendments. Test Article, Vehicle and/or Reagent Receipt, Preparation and Use. Animal Acquisition. Randomization Schedules. Mating History. Treatment (if prescribed by Staff Veterinarian). General Comments. Clinical Observations and/or General Appearance. Tissue and Sample Collection, Processing and Shipment. Cap and Labeled Tube Weights. Body Weights. Feed Consumption Values. Natural Delivery Observations. Litter Observations. Gross Necropsy Observations. Organ Weights (if required). Photographs (if required). Study Maintenance (room and environmental records). Feed, Water and Bedding Analyses. Packing and/or Shipment Lists. KEY PERSONNEL: Executive Director of Research: Mildred S. Christian, Ph.D., Fellow, ATS Director of Research: Alan M. Hoberman, Ph.D., DABT Associate Director of Research and Study Director Raymond G. York, Ph.D., DABT Director of Laboratory Operations: John F. Barnett, B.S. Manager of Study Coordination: Valerie A. Sharper, M.S. Manager of Animal Operations and Member, Institutional Animal Care and Use Committee: Dena C. Lebo, V.M.D. Manager of Regulatory Compliance: Barbara J. Patterson, B.A. Consultant, Veterinary Pathology: W. Ray Brown, D.V.M., Ph.D., ACVP 000414 418-014: PAGE C-20 Protocol418-014 Page20 FINAL REPORT: A comprehensive draft final report will be prepared on completion of the study and will be finalized following consultation with the Sponsor. The report will include the following: Summary and Conclusion. Experimental Design and Method. Evaluation of Test Results. Appendices: Figures, Summary and Individual Tables Summarizing the Above Data, Protocol and Associated Amendments and Deviations, Study Director's GLP Compliance Statement, Reports of Supporting Data (if appropriate) and QAU Statement. INSTITUTIONAL ANIMAL CARE AND USE COMMITTEE STATEMENT: The procedures described in this protocol have been reviewed by the Testing Facility's Institutional Animal Care and Use Committee. All procedures described in this protocol that involve study animals will be conducted in a manner to avoid or minimize discomfort, distress or pain to the animals. The Sponsor's signature below documents the fact that information concerning the necessity for conducting this study and the fact that this is not an unnecessarily duplicative study may be obtained from the Sponsor. No alternative (in vitro) procedures were available for meeting the stated purposes of the study. 000415 418-014:PAGE C-21 Protocol418-014 Page21 REFERENCES: 1. Christian, M.S. and Voytek, P.E. (1982). In Vivo Reproductive and Mutagenicity Tests. Environmental Protection Agency, Washington, D.C. National Technical Information Service, U.S. Department of Commerce, Springfield, VA 22161. 2. Christian, M.S. (1984). Reproductive toxicity and teratology evaluations of naltrexone (Proceedings of Naltrexone Symposium, 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 Charles River Cri:CDBR Rat. Charles River Laboratories, Inc., Wilmington, MA 01887-0630. (Data base provided by Argus Research Laboratories, Inc.) 4. Institute of Laboratory Animal Resources (1996). Guide for the Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 5. Salewski, E. (1964). Frbemethode zum makroskopischen Nachweis von Implantationsstellen am Uterus der Ratte. Arch. Pathol. Exp. Pharmakol. 247:367. 000416 PROTOCOL APPROVAL: FOR THE TESTING FACILITY Q 4/ - -- -- Alan M. Hoberman, Ph.D., DABT Director of Research Study Director 418-014: PAGE C-22 Protocol418-014 Page22 Date Z1- C .T -9 8 Date Dena C. Lebo, V.M.D. Member, Institutional Animal Care and Use Committee FOR THE SPONSOR ____________________ Marvin T. Case, D.V.M., Ph.D. Study Monitor Date A 7 o*J~ ?s' Date 000417 418-014:PAGE C-23 ATTACHMENT 1 SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE 000418 ATTACHMENT1 418-014: PAGE C-24 Protocol418-014 Page1of2 STUDY SCHEMATIC CROSS-FOSTERING STUDY" Start of Dosage Female Rats Premating Period (42 Days) End of Dosage Scheduled Sacrifice*- Cohabitation Period (5 days) Presumed Gestation Period Postpartum/ Lactation Period5 Dosage Period. a. For additional details see Tests, Analyses and Measurements" section of the protocol. b. F1 generation pups will be cross-fostered beginning on day 1 postpartum and continuing until day 21 postpartum. c. F1 generation pups will be sacrificed on day 21 postpartum. Fo generation dams will be sacrificed on day 22 postpartum. d. Samples for analysis of pharmacokinetics will be collected from six dams and their respective litters on day 14 postpartum. These litters will not be cross-fostered and will be sacrificed following sample collection. 000419 418-014: PAGE C-25 ATTACHMENT1 Protocol418-014 Page2of2 SCHEDULE* 27 OCT 98 Animals Arrive - Acclimation Begins. 02 NOV 98 Dosage Period - Female Rats (42 days prior to cohabitation until day 21 postpartum). 13 DEC 98 PM -19 DEC 98 AM 14 DEC 98 19 DEC 98 Cohabitation Period First Possible Day 0 of Presumed Gestation. Last Possible Day 0 of Presumed Gestation. 04 JAN 99 13 JAN 99 First Possible Delivery (Day 21 of presumed gestation). Last Possible Delivery (Day 25 of presumed gestation). 04 JAN 99 12 JAN 99 First Possible Day 1 Postpartum Culling Control Pool Only. Last Possible Day 1 Postpartum Culling Control Pool Only. 08 JAN 99 16 JAN 99 First Possible Day 4 Postpartum Culling Cross-Fostered Litters. Last Possible Day 4 Postpartum Culling Cross-Fostered Litters. 08 JAN 99 13 JAN 99 First Possible Day 25 of Presumed Gestation Female Sacrifice. Last Possible Day 25 of Presumed Gestation Female Sacrifice. 17 JAN 99 26 JAN 99 First Possible Day 14 Postpartum Pharmacokinetic Sacrifice. Last Possible Day 14 Postpartum Pharmacokinetic Sacrifice. 24 JAN 99 - 02 FEB 99 Scheduled Sacrifice - F1 Generation Pups (Day 21 postpartum). 25 JAN 99 - 03 FEB 99 Scheduled Sacrifice - Fo Generation Dams (Day 22 postpartum). 08 JUN 99 Draft Final Report. a. The study initiation date is the day the Study Director signs the protocol. 000420 418-014.PAGE C-26 ATTACHMENT 2 MATERIAL SAFETY DATA SHEET 000421 418-014: PAGE C-27 MATERIAL SAFETY DATA SHEET 3H 3M C enter 3 t . P aul, Hlnnaaota 55144-1000 1 . BOO-364-9577 o r (612) 737-6501 (24 hours) C o p yrig h t, 1998, H lnnaaota M inin g and M anufacturing Coapany. A il r ig h ts rasarvad. Copying and/or downloading o f th is in fo r a a tio n f o r th a purpoaa o f p ro p a rly u t i l i z i n g 3M p ro du cts is allowed provided th a t: 1) tha in f o r a a tio n i s copied in f u l l w ith no changes unless p r io r agreeaent i s o btained fr o a 3H, and 2) n e ith e r th e copy nor th e o r ig in a l i s re s o ld o r o th erw ise d is trib u te d w ith tha in te n tio n o f earning a p r o fit thereon. DIVISION: 3M CHEMICALS TRADE NAME: PC-9 5 FLU0RAD Brand F lu o ro c h e a ic a l S u rfa c ta n t ID NUMBER/U.P.C. : 98-0207-0103-7 00-51135-09054-1 98-0207-0104-5 98-0211-0888-5 00-51135-09362-7 98-0211-3916-1 Z F -0002-1044-1 ISSUED: January 29, 1998 SUPERSEDES: November 05, 1997 DOCUMENT: 10-3796-9 00-51135-09055-8 00-51135-02311-2 1 . INGREDIENT C.A.S. NO. PERCENT POTASSIUM PERFLUOROALKYL SULFONATE____ POTASSIUM PERFLUOROALKYL SULFONATE........ POTASSIUM PERFLUOROALKYL SULFONATE____ POTASSIUM PERFLUOROALKYL SULFONATE____ POTASSIUM PERFLUOROALKYL SULFONATE........ 2795-39-3 3871-99-6 29420-49-3 60270-55-5 3872-25-1 82 3 3 2 1 - 86 -S -7 -6 -3 2. PHYSICAL DATA BOILING POINT:........... VAPOR PRESSURE:......... VAPOR DENSITY:........... EVAPORATION RATE:. . . SOLUBILITY IN WATER: SPECIFIC GRAVITY:. . . PERCENT VOLATILE:. . . P H :.................................. V IS C O S IT Y :................... MELTING POINT:........... N/A N/A N/A N/A s lig h t ca. 0.6 Water=l (B u lk ) 0% 7- 8 (0.1% Aqueous) N/D N/D APPEARANCE AND ODOR: L ig h t colored, fre e flo w in g powder. 000422 Abbreviations: N/D - Not Deterained N/A - Not Applicable CA - Approximately 418-014.PAGE C-28 NSOS: FC-05 FLUORAD Brand F lu oroehem ical S u rfa c ta n t January 29, 1996 PAGE 2 3 . FIRE ANO EXPLOSION HAZARD DATA FLASH POINT:.........................................Nona FLAMMABLE LIMITS - LE L:..... N/A FLAMMABLE LIMITS UEL:..... N/A AUTOIGNITION TEMPERATURE:........... N/A EXTINGUISHING MEDIA: H a te r, Carbon d io x id e , Dry c h e a ic a l, Foaa SPECIAL FIRE FIGHTING PROCEDURES: Hear f u l l p ro te ctive c lo th in g , including hela e t, aelf-contained, p o s it iv e pressure o r p re ssu re deaand b re a th in g apparatus, bunker co a t and pants, bands around a ra , M a is t and le g s , fa ce aask, and p ro te c tiv e covering f o r exposed areas o f th e head. UNUSUAL FIRE AND EXPLOSION HAZARDS: See Hazardous D e co a p o sitio n s e c tio n f o r p ro d u cts o f co a b u stio n . 4 . REACTIVITY DATA STABILITY: Stable INCOMPATIBILITY - MATERIALS/CONDITIONS TO AVOID: Not applicable. HAZARDOUS POLYMERIZATION: Hazardous p o ly a e riz a tio n w i l l not o ccu r. HAZARDOUS DECOMPOSITION PRODUCTS: Carbon Monoxide and Carbon D io x id e , Oxides o f S u lfu r , Hydrogen F lu o rid e , T o xic Vapors, Gases o r P a r tic u la te s . 5 . ENVIRONMENTAL INFORMATION SPILL RESPONSE: Observe p re ca u tio n s fr o a o th e r s e c tio n s . Vacuum, use wet sweeping compound or w ater to a v o id d u s tin g . CAUTION! A vacuum c le a n e r c o u ld be an ig n it io n source. Clean up re sid u e w ith w a te r. Place in an approved metal co n ta in e r. Seal the co n ta in e r. RECOMMENDED DISPOSAL: Do not re le a s e to waterways o r sewer. Do n o t use in p ro ducts o r processes th a t could r e s u lt in aquatic concentrations g re a te r than 1 /1 0 o f the lo w e st EC50 o r LCSO c o n c e n tra tio n . In c in e ra te in an in d u s tr ia l or commercial f a c i l i t y in the presence o f a com bustible m a te r ia l. Combustion p ro d u c ts w i l l in c lu d e HF. D isp o sa l a lte r n a tiv e : Dispose o f waste product in a f a c i l i t y p e rm itte d to 000423 Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 418-014:PAGE C-29 MSOS: FC-95 PLUORAO Brand F lu o ro c h e a ic a l S u rfactan t January 29, 1998 5 . ENVIRONMENTAL INFORMATION (continuad) PAGE 3 accept cheaical waste. ENVIRONMENTAL DATA: 96-H r. Aquatic Fish LC50, Fathead Minnow(Piaephalea p ro a e la s)-3 8 ag/1, B lu e g ill S u n fis h (L e p o a is e a c ro c h iru s )*68 ag/1, Rainbow T ro u t(S a la o g a ird n e ri)* 1 1 mg/1; 4 8 -H r. EC50, Daphnia Magna * 50 a g /1 ; COO*.004 g / g ; BOO20 * N il . REGULATORY INFORMATION: V o la tile Organic Coepounds: N/A. VOC Less H20 & Exeapt S o lv e n ts : N/A. Since re g ula tio ns va ry, co n su lt applicable regulations or a u th o ritie s b e fo re d is p o s a l. U.S. EPA Hazardous Haste Nuaber * None (Not U.S. EPA Hazardous). T h is product coeplies w ith the cheaical re g is tra tio n requireeents of TSCA, EINECS, COSL, AICS, M ITI and Korea. EPCRA HAZARD CLASS: FIRE HAZARD: No PRESSURE: No REACTIVITY: No ACUTE: Yes CHRONIC: Yes 6. SUGGESTED FIRST AID EYE CONTACT: Ia m e d ia te ly flu s h eyes w ith la rg e aaounts o f water f o r a t le a s t 15 a in u te s . Get im nediate a e d ic a l a tte n tio n . SKIN CONTACT: Ia ie e d ia te ly flu s h s k in w ith la rg e aaounts o f w ate r. Reaove c o n ta a in a te d c lo t h in g . I f i r r i t a t i o n p e rs is ts , c a ll a p h y s ic ia n . Hash contaminated c lo th in g b e fo re reuse. INHALATION: I f signs/symptoms o c c u r, remove person to fre sh a i r . I f signs/symptoms c o n tin u e , c a ll a physician. IF SWALLOWED: D rin k two glasses o f w a te r. C a ll a p hysician. 7. PRECAUTIONARY INFORMATION EYE PROTECTION: A vo id eye c o n ta c t. Hear vented goggles. 000424 Abbreviations: N/D - Not Determined N/A - Not Applicable GA - Approximately 418-014: PAGE C-30 MSDS: FC-95 FLUORAO Brand F lu o ro c h e a ic a l S u rfa c ta n t January 29, 199S PAGE 4 7 . PRECAUTIONARY INFORMATION (continuad) SKIN PROTECTION: A vo id skin c o n ta c t. Hear a p p ro p ria te glovas when h a n d lin g t h is a t e r i a l . A p a ir o f glovas aada fr o a the fo llo w in g a a t e r ia l( s ) are racosaended: b u ty l ru b b e r. Use one o r aore o f th e fo llo w in g p e rso n a l p ro te c tio n ite a s as necessary to p revent s k in c o n ta c t: head covering, c o v e ra lls . P ro te c tiv e garaents (o th e r than gloves) should be aade o f e it h e r o f th e fo llo w in g a a te r ia ls : p o lyethylene/polyvinylidene ch lo rid e (Saranex). RECOMMENDED VENTILATION: Use w ith a p p ro p ria te lo c a l exhaust v e n t ila t io n . Use in a w e llv e n tila te d area. Provide s u ffic ie n t v e n tila tio n to s a in ta in e n is s io n s below recoasended exposure l i a i t s . I f exhaust v e n tila tio n i s not adequate, use a p p ro p ria te re s p ira to ry p ro te c tio n . RESPIRATORY PROTECTION: A v o id b re a th in g o f d u s t. S e le c t one o f the fo llo w in g NIOSH approved r e s p ira to r s based on a irb o rn e c o n c e n tra tio n o f co n ta s in a n ts and in accordance w ith OSHA r e g u la tio n s : h a lf-B a s k d u st and a is t r e s p ir a to r , half-M ask su p p lie d a i r re s p ira to r, f u ll- f a c e dust and a is t re s p ira to r, fu ll- fa c e supplied a ir re s p ira to r. PREVENTION OF ACCIDENTAL INGESTION: Do not e a t, d rin k o r ssoke when usin g t h is p ro d u c t. Hash exposed areas th o ro ug h ly w ith soap and w a te r. Hash hands a f t e r h a nd lin g and before eating. RECOMMENDED STORAGE: Keep c o n ta in e r d ry . Keep c o n ta in e r closed when n o t in use. FIRE AND EXPLOSION AVOIDANCE: N o n fla n m a ble . OTHER PRECAUTIONARY INFORMATION: No snoking: Ssoking w h ile using t h is product can r e s u lt in c o n ta a in a tio n of the tobacco and/or ssoke and lead to the fo rs a tio n o f the hazardous decoaposition products sentioned in se ctio n 4 o f t h i s MSDS. HMIS HAZARD RATINGS: HEALTH: 2 FLAMMABILITY: 0 REACTIVITY: 0 PERSONAL PROTECTION: X (See p re c a u tio n s , s e c tio n 7 .) EXPOSURE LIMITS INGREDIENT VALUE UNIT TYPE AUTH SKIN POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 POTASSIUM PERFLUOROALKYL SULFONATE.. . 0.1 MG/M3 POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3 POTASSIUM PERFLUOROALKYL SULFONATE.. . 0.1 MG/M3 TWA 3M TWA 3M TWA 3M TWA 3M Y Y Y 000425 Y Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 418-014:PAGE C-31 HSDS: FC-95 FLUORAD Brand F lu o ro c h e a ic a l 8 u rfa c ta n t January 29, 1998 PAGE 5 EXPOSURE LIMITS (continuad) INGREDIENT VALUE UNIT TYPE AUTM SKIN* POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/H3 TWA 3M Y * SKIN NOTATION: L is ta d substancaa in d ic a ta d w ith *Y' undar SKIN r a fe r to the p o te n tia l c o n tr ib u tio n to the o v e r a ll exposure by the cutaneous route in c lu d in g aucous aeabrana and aye, e it h e r by a irb o rn e o r, aore p a r t ic u la r ly , by d ir e c t c o n ta c t w ith th e substance. V e h ic le s can a lt e r s k in a b s o rp tio n . SOURCE OF EXPOSURE LIM IT DATA: - 3M: 3M Recoaaended Exposure G u id e lin e s 8 . HEALTH HAZARD DATA EYE CONTACT: M ild Eye I r r i t a t i o n : s ig n s /s y a p to a s can in c lu d e redness, s w e llin g , p a in , and te a r in g . SKIN CONTACT: M ild Skin I r r i t a t i o n ( a fte r prolonged o r repeated c o n ta c t): s ig n s /s y a p to a s can in c lu d e redness, s w e llin g , and itc h in g . May be absorbed th ro u g h th e s k in and p e r s is t in th e body f o r an extended tia e . INHALATION: May be h a r a fu l i f in h a le d . May be absorbed by in h a la tio n and p e r s is t in th e body f o r an extended tim e . S in g le overexposure, above recoaaended g u id e lin e s , may cause: I r r i t a t i o n (upper r e s p ir a t o r y ) : s ig n s /s y a p to a s can in c lu d e soreness o f th e nose and th r o a t, coughing and sneezing. IF SWALLOWED: Ing e stio n is not a lik e ly route o f exposure to th is product. I lln e s s aay r e s u lt fr o a a s in g le sw allo w in g o f a moderate q u a n tity o f th is m aterial. May be h a rm fu l i f sw allow ed. MUTAGENICITY: M u ta g e n ic ity assays in d ic a te th e pro du ct is not n u ta g e n ic. 000426 Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 418-014: PAGE C-32 MSDS: FC-95 FLUORAD Brand F lu o ro c h e a ic a l 8 u rfa c ta n t January 29, 1998 PAGE 6 8 . HEALTH HAZARD DATA (continued) REPRODUCTIVE/DEVELOPMENTAL TOXINS: N ot te ra to g e n ic in the r a t a t o r a l doaea below a a te m a lly to x ic le v e la . OTHER HEALTH HAZARD INFORMATION: T h ia p ro d u ct ia not known to c o n ta in any subatancea re g u la te d under C a lifo rn ia Proposition 65. A Product T o x ic ity Suaaary Sheet ia a v a ila b le . SECTION CHANGE OATES HEADING SECTION CHANGED SINCE Noveaber 05, 1997 ISSUE A b b re v ia tio n s : N/D Not D ete rnin e d N/A - Not A p p lic a b le GA - A p p ro x ia a te ly The in f o r a a tio n in th is M a te ria l S a fe ty Data Sheet (MSDS) i s b e lie v e d to be c o r r e c t as o f th e date is s u e d . 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 i s responsible f o r d e te ra in in g w hether th e 3M p ro du ct is f i t f o r a p a r t ic u la r purpose and s u ita b le f o r u s e r 's aethod o f use o r a p p lic a tio n . Given the v a r ie ty o f fa c to r s th a t can a f f e c t th e use and a p p lic a tio n o f a 3M p ro du ct, soee o f which are u n iq u e ly w ith in the u se r's knowledge and c o n tro l, i t is e s s e n tia l th a t th e u s e r e v a lu a te the 3M p ro d u c t t o determ ine whether i t is f i t f o r a p a r t ic u la r purpose and s u ita b le f o r u s e r's aethod o f use o r a p p lic a tio n . 3M p ro v id e s in fo r a a tio n in e le c t r o n ic fo r a as a s e rv ic e to i t s c u s to a e rs . Due t o the remote p o s s ib ilit y t h a t e le c tr o n ic tr a n s fe r aay have r e s u lte d in e r r o r s , o a is s io n s o r a lt e r a t io n s i n t h is in fo r a a tio n , 3M Bakes no re p re s e n ta tio n s as to i t s coapleteness o r accuracy. In a d d itio n , 00042V in fo rm a tio n o b ta in e d fr o a a database may not be as c u rre n t as the in fo rm a tio n in th e MSDS a v a ila b le d ir e c t ly fro a 3M. 418-014: PAGE C-33 ATTACHMENT 3 TEST ARTICLE PREPARATION PROCEDURE 000428 418-014.PAGE C-34 ATTACHMENT3 Protocol418-014 Version:418-014(22OCT981 Page1of2 TEST ARTICLE PREPARATION PROCEDURE Test Article: PFOS. Vehicle: 0.5% Tween 80 in R.O. Deionized Water. A. Purpose: The purpose of this procedure is to provide a method for the preparation of dosage suspensions of PFOS and the vehicle for oral administration to rats on Argus Study 418-014. B. General Information: 1. All suspension containers will be labeled and color coded. Each label will specify the protocol number, test article identification, Argus batch number, concentration, dosage level, preparation date, expiration date and storage conditions. 2. Suspensions will be prepared: X Daily __ Weekly For days of use Vehicle will be prepared: Dailv X Weekly For days of use 4. Suspensions will be prepared at a final dosage volume of 5 mL/kg. 5. Safety: X Gloves, lab coat, goggles or safety glasses and faceshield X Dust-Mist Respirator _ Half-Face Respirator _ Full-Face Respirator/Positive Pressure Hood _ Tyvek Suit/Apron 6. Dosage solutions adjusted for Free base and % Purity. __ Yes X No (Calculations based on 100%) __ Free Base __ Purity 7. Sampling requirements: Cited in protocol. 8. Storage: Cited in protocol. 000429 418-014: PAGE C-35 ATTACHMENT3 Protocol418-014 Version:418-014(2 2 OCT98) Page2of2 TEST ARTICLE PREPARATION PROCEDURE NOTE: Once the final volumes are achieved, stir bars are to be added to the containers; mixing should occur during sampling and/or administration. C. Preparation of Vehicle: 1. Add the required amount of R.O. deionized water to an appropriately labeled container. Heat the water to 50C, 5C, add the required amount of Tween 80 and mix until uniform. (See TEST ARTICLE CALCULATIONS for exact quantities.) D. Test Article Suspension Preparation: 1. To prepare the suspension for the high dosage group, add the required amount of test article (See TEST ARTICLE CALCULATIONS) into an appropriately sized, labeled container. QS ad to the required amount with vehicle, add a stir bar and heat the mixture to 80C, 5C, in a water bath for approximately 30 minutes or until the test article dissolves. 2. Once the test article has dissolved, remove the suspension from the water bath and spin the suspension while it cools. (Be sure there is a visible vortex, this will achieve the desired emulsion. This may be prepared the day before use.) Initial/Date : 000430 418-014: PAGE C-36 ATTACHMENT 4 CROSS-FOSTERING PROCEDURE 000431 418-014:PAGE C-37 ATTACHMENT4 CROSS-FOSTERING PROCEDURE Protocol418-014 Page1of1 The cohabitation period will begin one day earlier for the rats assigned to the control group. Dams assigned to the control group that mate on the first day will be allowed to deliver and will retain their natural pups until needed for cross-fostering purposes. This will provide a pool of control dams available for immediate cross-fostering to pups from PFOS-treated dams, thus preventing these pups from nursing from their PFOS-treated dams. On the remaining days of cohabitation, both control and PFOS-treated groups will be co-housed with male breeders, one male rat to one female rat. Starting with this set of deliveries, all pups will be removed immediately (as soon as parturition is complete) from their respective dams and placed with either another dam assigned to the control group or a PFOS-treated dam. If a dam assigned to the control group is not available, then a dam from the control pool will be used. The litter from that control pool dam will then be sacrificed via decapitation. The first ten pups from the control pool (irrespective of litter) determined to be at day 1 postpartum will be decapitated, and the lungs (saved in Bouin's solution) and the liver (saved in neutral buffered 10% formalin) will be collected and preserved for possible future histopathological evaluation. The remaining pups from the control pool dams will be sacrificed via decapitation and discarded without necropsy evaluation. This cross-fostering procedure will result in four groups of 12 dams/pups, i.e., Control/Control (Subset A), Control/PFOS (Subset B), PFOS/Control (Subset C) and PFOS/PFOS (Subset D). This procedure will allow distinctions to be made between prenatal and postnatal effects of the continuous maternal PFOS treatment. 000432 418-014: PAGE C-38 O Primedica Aig9u0s5TReSelThseeepeealhehroHcfynahoxeDrL::srah((ib22va1o1em,55ra,))BtPo44uA4r4iil33ed--1si88n,957g0I1n84Ac047. PROTOCOL 418-014 Oral (Gavage) Cross-Fostering Study of PFOS in Rats SPONSOR'S STUDY NUMBER: T-6295.13 Amendment 1 - 4 January 1999 1. Cross-Fostering Procedure (Paragraph 3 of Attachment 4 to the protocol): The first ten pups from the control pool (irrespective of litter) and the first ten pups that are found dead (no autolysis present) per dosage group determined to be at day 1 postpartum will be decapitated at approximately 1 to 3 hours after birth, and the lungs and the liver will be retained. Several (minimum of two) sections (approximately 1 mm thick) of the lung and the liver will be removed using a scalpel and will be retained in McDowell-Trump's Fixative [preparation procedure provided by the Sponsor is documented in the raw data (stored under refrigeration in scintillation vials) (see attached)] for future evaluation. The remaining lung and liver samples will be retained in Bouin's solution or neutral buffered 10% formalin, respectively, for possible future evaluation. The remaining pups from the control group dams will be sacrificed via decapitation and discarded without necropsy evaluation. The liver sections in McDowell-Trump's Fixative will be shipped (on cold packs), for evaluation by electron microscopy, to: Jeanne deWard Pathology Associates International 4915 D Prospectus Drive Durham, North Carolina 27713 Telephone: (919)544-5257 Telefax: (919)544-3218 The remaining samples will be shipped (ambient conditions) to the Kris J. Hansen, Ph.D., at the address cited in the protocol for histopathological and biochemical evaluation. 000433 418-014:PAGE C-39 Protocol418-014 Amendment 1 Page2 Reason for Change: The Sponsor has requested that these additional samples be retained for continued evaluation of the reasons for reduced pup survival observed in previous studies. 2. Cross-Fostering Procedure (Paragraph 4 of Attachment 4 to the protocol): The fourth paragraph of the Cross-Fostering procedure has been changed to read: This cross-fostering procedure will result in four 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 will allow distinctions to be made between prenatal and postnatal effect of the continuous maternal PFOS treatment. Reason for Change: This corrects the protocol. CU - ' Alan M. Hoberman, Ph.D., DABT Director of Research Date ind G. York, Ph.., DABT Date Associate Director orResearch and Study Director Dena C. Lebo, V.M.D. Date Chairperson, Institutional Animal Care and Use Committee Marvin T. Case, D.V.M., Ph.D. Study Monitor Date 000434 418-014:PAGE C-40 Protocol418-014 Version:418-014(2 9 Dec981 Page1of2 FIXATIVE PREPARATION PROCEDURE Fixative : McDowell-Trump's Fixative A. Purpose: The purpose of this procedure is to provide a method for the preparation of McDowell-Trump's Fixative for tissue samples for study 418-014. B. General Information: 1. All solution containers will be labeled. Each label will specify the protocol number, fixative identification, preparation date, expiration date and storage conditions. 2. Fixative will be prepared: _ Daily _ Weekly __Once 3. Safety: X Gloves, lab coat, goggles or safety glasses and face shield X Dust-Mist Respirator X Half-Face Respirator _ Full-Face Respirator/Positive Pressure Hood _ Tyvek Suit/Apron X Other: Chemical Fume Hood 4. Dosage suspensions adjusted for % Activity/Purity or Correction Factor: __ Yes X No (Calculations based on 100%) 5. Sampling requirements: Cited inprotocol. 7. Storage: Room temperature 000435 418-014: PAGE C-41 Protocol 418-014 Version:418-014(29Dec981 Page2of2 FIXATIVE PREPARATION PROCEDURE C. Preparation of Fixative: McDowell-Trump's Fixative 1. Add 860 mLs of R.O. Deionized water into an appropriately labeled container. 2. Add 100 mLs of 37% Formaldehyde to the container and stir until uniform. 3. Add 40 mLs of Gluteraldehyde to the container and stir until uniform. 4. Add 1 1.6g o f Sodium Phosphate Monobasic to the container and stir until uniform. 5. Add 2.70g o f Sodium Hydroxide to the vessel and stir until uniform. [NOTE: The final solution should have a pH o f approximately 7.3 ] Written By: -- -V- Clarification: y N o __Yes [see attached clarification form] Initials/Date: G00436 APPENDIX D DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 000437 APPENDIX E TEMPERATURE AND RELATIVE HUMIDITY REPORTS 000438 418-014: PAGE E-1 ARGUS Tem perature and Relative Hum idity R eport Location: Room 04 Protocol Number: 418-014 Range o f D ates: 27-O ct-1998 13:15 to 23-N ov-1998 15:49 Target Range: Species: Rat Total Number of Days: Total Number of Hours: Total Number of Data Points: Temperature 64#F to 79*F 28 650.51 634 Relative Humidity 30% to 70% 28 650.51 634 Mean ( SD): Maximum: Median: Minimum: Number of Points in Range (%): Number of Points High (%): Number of Points Low (%): 69.1 (1.9) 53.2 (8.0) 76.4 66.2 68.4 57.2 66.5 28.6 634 (100.0) 633 0 (0.0) 0 0 (0.0) 1 (99.8) (0.0) (0.2) Report Generated: 04-Feb-1999 at 17:26 COMMENTS: REVIEWED BY: DATE: t-SW C u m u la tiv o hw I f\tf\A fM T\ 00439 418-014:PAGE E-2 ARGUS Temperature and Relative Humidity Report Location: Room 27 Protocol Number: 418-014 Range of Dates: 23-Nov-1998 15:49 to 31-Dec-1998 14:10 Target Range: Species: Rat Total Number of Days: Total Number of Hours: Total Number of Data Points: Temperature 64*F to 79*F 39 910.01 913 Relative Humidity 30% to 70% 39 910.01 913 Mean ( SD): Maximum: Median: Minimum: Number of Points in Range (%): Number of Points High (%): Number of Points Low (%): 70.9 ( 0.7) 57.1 (6.1) 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 at 17:28 COMMENTS: REVIEWED BY:_________tyl_`TtbjlL DATE: j.t 7? 000440 418-014:PAGE E-3 ARGUS Temperature and Relative Humidity Report Location. Room 35-37 Protocol Number: 418-014 Range of Dates: 31-Dec-1998 14:10 to 18-Jan-1999 08:45 Target Range: Species: Rat Total Number of Days: Total Number of Hours: Total Number of Data Points: Temperature 64'F to 79*F 19 426.24 427 Relative Humidity 30% to 70% 19 426.24 427 Mean ( SD): Maximum: Median: Minimum: Number of Points in Range (%): Number of Points High (%): Number of Points Low (%): 75.6 (1.7) 50.1 (7.4) 78.3 56.1 76.4 51.6 68.5 5.0 427 (100.0) 417 0 (0.0) 0 0 (0.0) 10 (97.7) (0.0) (2.3) Report Generated: 04-Feb-1999 at 17:30 COMMENTS: REVIEWED B Y :_____________ % . / T f e f l L ____________ DATE: l - f i H 000441 418-014: PAGE E-4 ARGUS Temperature and Relative Humidity Report Location: Room 27 Protocol Number: 418-014 Range of Dates: 18-Jan-1999 08:45 to 29-Jan-1999 09:57 Target Range: Species: Rat Total Number of Days: Total Number of Hours: Total Number of Data Points: Temperature 64'F to 79#F 12 265.0 266 Relative Humidity 30% to 70% 12 265.0 266 Mean (1 SD): Maximum: Median: Minimum: Number of Points in Range (%): Number of Points High (%): Number of Points Low (%): 70.4 ( 0.5) 54.3 (8.3) 72.0 70.4 70.4 56.4 69.1 33.1 266 (100.0) 265 0 (0.0) 1 0 (0.0) 0 (99.6) (0.4) (0.0) Report Generated: 04-Feb-1999 at 17:33 COMMENTS: REVIEWED BY: n i.U l O .- DATE: 1 G00442 418-014: PAGE E-5 ARGUS Relative Humidity Deviations Report 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 Time 03-Nov-1998 14:00 R.H. 28.6L 30% to 70% Date Time R.H. H=Valueout ofrange-High L=Valueout ofrange-Low R.H. * RelativeHumidity(%) Report Generated: 13-May-1999at 10:45 Thesedeviationsdidnot adverselyaffecttheoutcomeorinterpretationofthestudy. ____Thefollowingdeviation(s) impactedontheoutcomeofthestudyasdescribed: DeviationsbvLocation h /04 m Q7t 000443 418-014: PAGE E-6 ARGUS Relative Humidity Deviations Report Location: Room 35-37 Protocol Number: 418-014 Range of Dates: 31-Dec-1998 14:10 to 18-Jan-1999 08:45 Humidity Target Range: Species: rat Date 05-Jan-1999 05-Jan-1999 05-Jan-1999 06-Jan-1999 06-Jan-1999 06-Jan-1999 06-Jan-1999 06-Jan-1999 06-Jan-1999 06-Jan-1999 Time 21:00 22:00 23:00 00:00 01:00 02:00 03:00 04:00 05:00 06:00 R.H. 5.8L 5.1 L 5.0L 5.1 L 5.1 L 5.2L 5.3L 5.5 L 5.6L 5.6L 30% to 70% Date Time H=Valueout ofrange-High L=Valueout ofrange-Low R.H. =RelativeHumidity(%) Report Generated: 13-May-1999at 10:50 y Thesedeviationsdidnot adverselyaffect theoutcomeorinterpretationofthestudy. ______Thefollowingdeviation(s) impactedontheoutcomeofthestudyasdescribed: I n rj fin n f \ j ( \ A m Q"7\ 000444 418-014: PAGE E-7 ARGUS Relative Humidity Deviations Report 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 Time 24-Jan-1999 06:00 R.H. 70.4H 30% to 70% Date Time H=Valueout ofrange-High L=Valueout ofrange-Low R.H. =RelativeHumidity(%) Report Generated: 13-May-1999at 10:54 Thesedeviationsdidnot adverselyaffecttheoutcomeorinterpretationofthestudy. Thefollowingdeviation(s) impactedontheoutcomeofthestudyasdescribed: n r s t i r tn ( \ t f \ A fM OTA 000445 418-014:PAGE D-1 DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 1. Two extra female from the treated group (Group II) were used for pharmacokinetic sample collection on day 14 of lactation (DL 14), 22 January 1999. These two female rats in turn received all collections and were sacrificed on DL 14. This deviation did not adversely affect the outcome of the study because the pharmacokinetic samples were not analyzed. 2. Eight female rats rather than six female rats from the control group (Group I) were used for pharmacokinetic sample collection on DL 14,17 January 1999, 21 January 1999 and 22 January 1999. The two extra female rats received all sample collections and were sacrificed on DL 14. This deviation did not adversely affect the outcome of the study because the pharmacokinetic samples were not analyzed. 3. A total of 25 litters from the control (Group I) and treated (Group II) groups were cross-fostered rather than 24 litters. This deviation did not adversely affect the outcome of the study because only one additional litter was added to each dosage group. 4. Twenty-five female rats rather than all of the rats in the control group (Group I) were placed into cohabitation on 13 December 1998. The cohabitation period for these rats lasted six days rather than five days. This deviation did not adversely affect the outcome of the study because it provided sufficient mated rats for cross-fostering. All deviations are documented in the raw data. 000446 APPENDIX F PATHOLOGY REPORT 000447 418-014:PAGE F-1 Pathology Associates international A Company of Science Applications international Corporation PATHOLOGY REPORT (ANCILLARY STUDY) ELECTRON MICROSCOPIC EVALUATION OF LIVER AND LUNG IN CRL:CDBR VAF/PLUS RATS) ORAL (GAVAGE) CROSS-FSTERING STUDY OF PFOS IN RATS CLIENT STUDY NUMBER 418-014 PAI STUDY NUMBER EM 99.46 Prepared by: James B. Nold, D.V.M., Ph.D., Diplomate, A.C.V.P. Pathology Associates International 4915-D Prospectus Drive Durham, NC 27713 April 5 ,1 9 9 9 PREPARED FOR 3M CORPORATE TOXICOLOGY, 3M CENTER, ST. PAUL, MN 55144-1000 000448 4915DProspectusDriveDurham,NorthCarolina 27713(919) 544-5257(919) 544-3218FAX 418-014:PAGE F-2 Pathology Narrative Transmission Electron Micrograph Interpretation Forms Transmission Electron Micrographs Quality Assurance Statement Section I II in IV 000449 418-014:PAGE F-3 Pathology Narrative 000450 418-014P A G E F-4 PnmedicaAAncriglluasryStPuadtyhoNloog. y41R8e-p0o1r4t PATHOLOGY REPORT (ANCILLARY STUDY) ELECTRON MICROSCOPIC EVALUATION OF LIVER AND LUNG IN CRL:CDBR VAF/PLUS RATS) ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS CLIENT STUDY NUMBER 418-014 PAI STUDY NUMBER EM 99.46 SUMMMARY Ultrastructurally, there were increased numbers o f peroxisomes within hepatocytes of neonatal rat pup livers from dams treated with 1.6 mg/kg/day (high-dose) PFOS compared with controls. Quantitation of peroxisomes indicated slightly more than twice the number o f peroxisomes in treated animals. Differences in cellular membranes or mitochondria were not discernible between control and treated animals in the samples examined. Type II pneumocytes containing lamellar bodies were identified in both controls and high-dose animals; however, there appeared to be an increased number of type II pneumocytes and lamellar bodies in lungs from treated samples. Although some lamellar material (surfactant) was identifiable within alveolar lumina, no significant difference was noted between control and treated samples. PROCEDURES The objective o f this study was to examine by electron microscopy the livers and lungs from selected day-1 postpartum rat pups in an oral (gavage) cross-fostering study of PFOS. In addition to a general qualitative evaluation for differences between control and treated animals, the ultrastructural endpoints included quantitation o f peroxisomes within hepatocytes, membrane and mitochondrial changes in hepatocytes, and the quantitation of surfactant lamellar bodies in the lung. For the cross-fostering study, 6 6 female Crl:CDBR VAF/PLUS rats were given the test article or the vehicle once daily beginning 4 2 days prior to cohabitation with a male through day 21 postpartum according to Text Table 1. Text Table 1. Dosage Levels, Concentrations and Volumes DGorsoaugpe I NuFmebmearloefRMatasted 30 + 6* P(FmOgS/kgD/doasayg)e 0 (Vehicle) PFOS (mCogn/mceLn)tration 0 Dos(amgeLV/kogl)ume 5 n 24 + 6a 1.6 0.32 5 a S1a4mppolsetpsafrotruman.alTyhsiesseofdpahmasrmanadcotkhieniretriecsspwecetrievecoUlltteecrtsewd efrroemsa6crdifaimcesdafnodllothweiinrgressapmecptlievecoliltlteecrtsioonn. day 000451 i-i 418-014:PAGE F-5 PrimedicAa AncriglluasryStPuadtyhoNloog.y41R8e-p0o1r4t The first ten pups from the control pool (irrespective o f litter) and the first 10 pups that were found dead (no autolysis present) from the treated group determined to be at day I postpartum were decapitated at approximately 1-3 hours after birth, and the lungs and liver were collected. Samples o f lung and liver were thin-sliced and placed in M cDowellTrump fixative (M cDowell EM, 1976), and submitted to this laboratory for electron microscopy processing and evaluation. Per sponsor request, tissue samples from selected pups were processed and evaluated ultrastructurally (Text Table 2). Pup numbers were assigned by this laboratory because submitted samples were identified by dam number only. The five digit number is the dam number; the suffix 1 or 2 denotes the pup number from that dam's litter. Text Table 2. Animals Selected for Electron Microscopy I Control (0 mg/kg/day) 1111188888999991100152445-----11122 High-Dos1e1118888(999915755.66322----m1112g/kg/day) The lungs and livers were then processed into Spurr's resin for ultrastructural examination. Thick (lp.) sections were cut, stained with toluidine blue, and examined to select representative areas for further electron microscopy processing. Thin sections (approximately 9 0 nm) were cut, mounted on 200-m esh copper grids, stained with 5 % methanolic uranyl acetate and Reynold's lead citrate, and examined on a Zeiss 9 0 0 transmission electron microscope. Centrilobular hepatocytes, where clearly identifiable in liver sections, were preferentially examined. Representative electron photomicrographs o f liver and lung were taken and significant ultrastructural features were summarized for each photograph and animal on a designated transmission electron micrograph interpretation form. The number o f peroxisomes in hepatocytes was manually counted for each center photographed hepatocyte and recorded. RESULTS AND DISCUSSION Individual interpretations o f light and electron micrographs for each animal are shown in Section H. Selected representative electron photomicrographs were labeled and are shown in Section EH. Fixation o f the tissues was less than optimal for electron microscopy. This was especially evident in the liver compared with the lung. Lung is inherently less dense than liver and allows better infiltration o f the fixative into the tissue. Suboptimal fixation was characterized by discontinuous plasma membranes, loss o f detail o f cytoplasmic organelles, dilatation o f endoplasmic reticulum and the nuclear envelope, and dilatation o f mitochondrial cristae. Fixation, however, was still considered adequate to assess for 000452 1-2 418-014:PAGE F-6 PrimedicaAAncriglluasryStPuadtyhoNloog. y41R8e-p0o1r4t the presence o f peroxisomes in the hcpatocytes and lamellar bodies in the lung and to make general comparisons between control and treated samples. LIVER Control (0 mg/kg/day). Subcellular features o f control hepatocytes are demonstrated in Figures 1 and 2. Hepatocytes characteristically contain numerous mitochondria and prominent endoplasmic reticulum. In these cells rough endoplasmic reticulum was evident, but smooth endoplasmic reticulum was not because o f suboptimal fixation. Mitochondria generally appeared dark and contained scattered dense deposits and some dilated cristae, both artifacts due to suboptimal fixation. Dilatation of endoplasmic reticulum and the nuclear envelope were additional artifacts o f poor fixation. Fine granular material in the cytoplasm is consistent with glycogen particles. Peroxisomes were identified and quantitated in from representative hepatocytes. For the control livers, the average number o f peroxisomes counted per hepatocyte photograph was 7 peroxisomes per cell (Text Table 3). Fetal and neonatal rat liver is a hematopoietic tissue, and numerous hematopoietic cells were evident within the liver sinusoids. | Text Table 3. Quantitation of Hepatocellular Peroxisomes 1 Control Anim1111188888a99999l11100N52454-u----11122mber (0Nmugm/pkbgeer/rdHoafye49668)Pp.....48602aetroocxyisteomes Mean Control = 7.0 High-Dose (1.6 mg/kg/day) Anim11118888a9999l5755N2326u----112m1 ber Numpbeerr HofePpaetroocxvisteomes 11117674.2...0005 Mean High-Dose = 16.1 H igh-Dose (1.6 mg/kg/day). The average number of peroxisomes per hepatocyte (14.7) was higher in the high-dose animals compared with controls (Text Table 3). Otherwise, general ultrastructural morphology was similar in high-dose livers compared with control livers. Figures 3-5 illustrate representative hepatocytes from several high-dose animals. Differences in mitochondria and cellular membranes were not discernible between highdose and control livers. However, suboptimal fixation of the tissue precluded detailed evaluation of these organelles. LUNG C ontrol (0 m g/kg/day). Lung samples were atelectatic and not inflated with fixative. Therefore most alveoli and alveolar septa were collapsed with poor delineation o f alveolar and airway spaces. Figures 6-8 illustrate some normal cellularity and ultrastructural pulmonary anatomy. Type II pneumocytes (PN 2) were infrequently identified lining alveoli, terminal bronchioles, or within collapsed septa. Type II pneumocytes, were identified by the presence o f lamellar bodies, which are cytoplasmic 000453 1-3 418-014:PAGE F-7 PrimedicaAAncriglluasryStPuadtyhoNloog. y41R8e-p0o14n vacuoles containing electron-dense laminated membrane-like profiles. Lamellar bodies represent the material secreted into the alveolar spaces as surfactant. Surfactant or lamellar material within airway spaces was very rarely identified in control lungs. The type I pneumocyte (PN 1) is characteristically flattened and lines the alveolar space as a thin margin o f cytoplasm. Type I pneumocytes differentiate from type II pneumocytes. Many pneumocytes appeared to be cuboidal without distinctive lamellar bodies, and may represent a transition stage between PN2s and P N ls, especially since these lungs were neonatal and had not been fully expanded with air. Additionally, some cuboidal epithelial cells represented terminal bronchiolar epithelium. Type II pneumocytes and lamellar bodies were infrequently identified in these control lungs and no reliable quantitation could be assessed o f them. Some lamellar-like bodies were also present within capillary lumina. High-Dose (1.6 mg/kg/day). Ultrastructural pathology o f the high-dose lungs was essentially similar to control samples. There appeared, however, to be increased numbers o f type II pneumocytes and lamellar bodies; at least, identifying them for illustrative purposes was easier than in control samples. The collapsed nature of most o f the alveoli and septa made definitive quantitation impractical. Some lamellar bodies were identified within capillary lumina and only a small number were seen within alveoli. Figures 9-12 illustrate some representative lung fields from the high-dose animals. CONCLUSION Ultrastructural evaluation o f liver and lungs of rat pups from dams treated with 0 (control) or 1.6 mg/kg/day (high-dose) PFOS identified increased numbers of peroxisomes within hepatocytes o f high-dose rat pup livers. Quantitation o f peroxisomes indicated slightly more than twice the number of peroxisomes in treated animals. Differences in cellular membranes or mitochondria were not discernible between control and treated animals in the samples examined. In lung samples the presence o f type II pneumocytes and lamellar bodies was evaluated. Type II pneumocytes containing lamellar bodies were identified in both controls and high-dose animals; however, there appeared to be an increased number o f type II pneumocytes and lamellar bodies in lungs from treated samples. Some lamellar bodies and lamellar myelin-like figures were extracellular and within capillary lumina. Although some lamellar material (surfactant) was identifiable within alveolar lumina, no significant difference was noted between control and treated samples. SIGNATURE OF AUTHOR Diplomate, A.C.V.P. 1-4 000454 418-014PAG E F-8 PrimedicAa AncriglluasryStPuadtyhoNloog.y41R8e-p0o14n REFERENCES M cDowell, EM and Trump, BF: Histologic fixative for routine diagnostic light and electron microscopy. Arch. P athol. Lab. M ed., 100:405-414, 1976. 000455 1-5 418-014.PAGE F-9 II. Transmission Electron Micrograph Interpretation Forms1 1Forms are typographically corrected versions o f signed/dated raw data sheets maintained in archived study file. 000456 418-014:PAGE F-10 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014: EM -99.46 Animal No.: 18904-1___________ Treatment Group: 0 me/ke Control Sex: non applicable_________ Species/Strain: CRarkt CDOBR v a f /PIus Tissue: Liver ______________ Block No(s).: 99.46-1__________ Z15683, Z15684, Photo/Negative No(s).: Z 15686-88 Significant Lesions (check one): _________Yes X No Interpreting Pathologist (signature and date): ___________________ Features: Z 15683:- Hepatocyte, several erythrocytes in sinusoid, and nucleus o f probable endothelial cell on right side o f photograph. Six (6) peroxisomes evident in large central hepatocyte. Dilated blebs in RER are pseudo-peroxisomes. Abundant fine granular material, glycogen, in cytosol. Dense mitochondria with some granular deposits and dilated cristae. Poor definition o f endothelial cells. Z 15684: Several hepatocytic nuclei present, but poor cytoplasmic margin definition. Some swelling of mitochondria. Dilated blebs in RER. Bile canaliculus in lower left comer. Two (2) peroxisomes in central cell. In adjacent two cells only single peroxisomes are clearly evident. Z 15686: Single hepatocyte surrounded by erythrocytes. No clear definition of endothelial cells. Distended cytosol with glycogen-like granular particles. Abundant mitochondria and RER. Mitochondria have some granular densities and dilated cristae. Dilated blebs in RER. Ten (10) peroxisomes evident. Z 15687: Single hepatocyte. Granulocytic cell (hematopoietic) present in bottom left comer. Distended cytosol with granular material. Dense mitochondria with some granular deposits. Two (2) peroxisomes evident in central hepatocyte. Z 15688: Hepatocyte. Erythroid precursor present in lower left sinusoid. Dilated blebs in RER. Cytosol expanded with fine granular material, probably glycogen. Three (3) peroxisomes present. Conclusions: Normal hepatocytes. 23/5 = 4.6 peroxisomes per hepatocyte evident. Plasma membranes poorly defined. Dense mitochondria with some granular deposits and dilated cristae. Suboptimal fixation._________________________________________________ 000457 418-014:PAGE F-11 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014: EM -99.46 Animal No.: 18904-1__________ Treatment Group: 0 me/kg Control Sex: non applicable_________ Species/Strain: Crl:CDBR VAF/Plus -El Tissue: Lung_________________ Block No(s).: 99.46-2_________ Z15689 - Photo/Negative N o (s ).: _ Z 1 5 6 9 3 _ _ Significant Lesions (check one): _________Yes X No Interpreting Pathologist (signature and date): ___________________ Features: Z 15689: Capillary with three endothelial cell nuclei. In lumen o f capillary is a lamellar-like body. Several type 1 pneumocytes (PN1) present surrounding capillary. N o pneumocyte lamellar bodies present. Z 15690: Type 2 pneumocyte (PN2) present, containing multiple lamellar bodies (-7). Beneath the PN2 is a capillary containing an erythrocyte, platelet, and endothelial cell nuclei. PN1 to right side o f photograph with more translucent cytoplasm. Z 15691: Collapsed alveolar septa. On top is a PN1 with flattened cytoplasm and large round nucleus. Other cells less clearly defined, but appear to be primarily P N ls in collapsed septa. N o pneumocyte lamellar bodies evident. Z 15692: Collapsed alveolar septa. On top is a thin-layered PN1, and in the bottom left is larger profile o f a PN1. Centrally appears to be a capillary lumen containing some lamellar debris and two leucocytes. Also some lipid debris is present in the capillary lumen. Endothelial cell cytoplasm is evident, but no endothelial cell nucleus. No pneumocyte lamellar bodies identified. Z 15693: Collapsed alveolar septa. At top is a profile o f a P N 1 in the alveolar space. Beneath it is a thin-layered PN1, and to the left side o f photo is a PN1. Subjacent nuclei are probably endothelial cell nuclei. At bottom left is an erythrocyte. Just to the right is a collapsed alveoli and thin-line PN1. No pneumocyte lamellar bodies identified. Conclusions: Normal lung tissue. Fixation adequate. Only one PN2 identified. 000458 418-014:PAGE F-12 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014: EM -99.46 Animal No.: 18904-2___________ Treatment Group: 0 mg/ke/ Control Sex: non applicable_________ Species/Strain: CRralt:CD<g)BR v a f /pius Tissue:_____ Liver ____________ Block No(s).: 99.46-3___________ Z15694 - Photo/Negative N o (s ).: _ Z 1 5 6 9 8 _ _ Significant Lesions (check one): _________Yes X No Interpreting Pathologist (signature and date): ___________________ Features: Z 15694: Hepatocyte. Multiple mitochondria, some RER and multiple rounded blebs o f RER. Cytoplasm full o f granular glycogen. Poor endothelial cell definition in sinusoids. Nucleated erythrocyte in bottom left. Mitochondria appear dense with some granular deposits and dilated cristae. Three (3) peroxisomes identified. Z 15695: Hepatocyte. Dilated RER. Multiple dense mitochondria with granular deposits and dilated cristae. Cytoplasmic margins and endothelial cells poorly defined. Six (6) peroxisomes identified. Z 15696: Hepatocyte. Many dilated blebs of RER. Dense mitochondria with granular deposits and dilated cristae. Top left hepatocyte has swollen, less dense, mitochondria. Twelve (1 2 ) peroxisomes identified. Z 15697: Several poorly-fixed hepatocytes. Many light and dark dilated blebs o f RER, which are difficult to differentiate from peroxisomes. Five (5) peroxisomes identified in single more-circumscribed hepatocyte. Z 15698: Hepatocyte. Large clear areas in cell contain fine granular material consistent with glycogen. Five (5) peroxisomes evident. Conclusions: Normal hepatocytes. Suboptimal fixation. 31/5 = 6.2 peroxisomes per cell. 000459 418-014:PAGE F-13 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014: EM -99.46 Animal No.: 1 8 9 0 4 - 2 _________ Treatment Group: 0 mg/kg Control Sex: non applicable_________ Species/Strain: CitCDOBR v a f /p 1us<s> Rat Tissue: Lung ______________ Block No(s).: 99.46-4__________ 15699 - Photo/Negative Nofs).: Z 15704 Significant Lesions (check one): _________Yes X No Interpreting Pathologist (signature and date): ___________________ Features: Z 15699: Alveolus containing several cuboidal pneumocyte cells. The lower cell appears to contain a couple small lamellar bodies, probably a PN2. The other two large cuboidal cells do not contain any lamellar bodies, but appear to be pneumocytes which haven't flattened out into P N ls or are terminal bronchiolar epithelia. At the left and bottom are capillaries lined by endothelial cells. The flattened PN1 at the bottom contains a segment o f luminal surface with some dense electron-dense material. Some dilated cristae in mitochondria. Z 15700: Alveolar space with capillary/erythrocyte at left and pneumocytes around top and right. Top left pneumocyte contains three "empty" vacuoles, possibly extruded lamellar bodies. PN1 at right is slightly more electron dense. Z 15701: Multiple P N ls lining alveolar spaced. N o lamellar bodies or PN2s evident. Capillary at lower right with endothelial cells. Z 15702: Cell type containing central lamellar body not clearly discernible. It appears to be outside the capillary and endothelial cell at top left, but compressed between the pneumocyte on right and lower central cell. This is an area o f collapsed alveolar septa. Z 15703: Alveolus and lining cells. Large PN1 appears to contain a single lamellar body in center o f photograph. Capillary and endothelium at left o f photograph. Collapsed alveolar septa in bottom center and right. Dilated cristae in some mitochondria. Z 15704: Alveolus with lining pneumocytes. Cell in lower left comer contains partiallyempty vacuole containing some lamellar material. Denser cell at center left contains a couple clear vacuoles, although this cell is lined by a flattened PN1.___________________ Conclusions: Normal lung and alveoli. A few lamellar bodies identified. 000460 418-014:PAGE F-14 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014: EM -99.46 Animal No.: 18912-1__________ Treatment Group: 0 me/kg. Control Sex: non applicable_________ Species/Strain: Crl:CD<g)BR VAF/Pius(S> Rat Tissue:_____ Liver_______________ Block No(s).: 99.46-5__________ Z15773 - Photo/Negative No(s).: Z 15777 Significant Lesions (check one): _________Yes X No Interpreting Pathologist (signature and date): ___________________ Features: Z 15773: Hepatocyte. Some segments of hematopoietic cells are present in the upper right comer. Endothelial cell details are indistinct, and fixation is poor. There is dilation and blebbing of the RER and some dilation of the nuclear envelope. Six (6) peroxisomes counted. Z 15774: Hepatocyte surrounded by some hematopoietic cells. Marked dilation and blebbing of RER. Four (4) peroxisomes counted. Z 15775: Hepatocyte with canaliculus at the bottom and sinusoidal erythrocyte at top right. Endothelial cells poorly-defined and Fixation is suboptimal. Eleven (1 1 ) peroxisomes counted. Z 15776: Hepatocyte is poorly-fixed and disrupted. Marked dilation and blebbing of the RER. Mitochondria are dense and contain some granular deposits and dilated cristae. Seven (7 ) peroxisomes counted. Z 15777: Hepatocyte appears similar to those described above. It is poorly fixed and RER and nuclear envelope are dilated. Seventeen (17) peroxisomes counted. Conclusions: Suboptimal fixation. Nine (9 ) peroxisomes/hepatocyte (average o f 5 cells) 000461 418-014:PAGE F-15 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014: EM -99.46 Animal No.: 18912-1___________ Treatment Group: 0 me/kg. Control Sex: non applicable_________ Species/Strain: CRralt:CD<3>BR VAF/Plus Tissue: Lung___________________ Block No(s).: 99.46-6_________ Z15768 - Pboto/Negative No(s).: Z 15772 Significant Lesions (check one): _________Yes X No Interpreting Pathologist (signature and date): ___________________ Features: Z 15768: A row of cuboidal epithelial cells, probably bronchiolar epithelium. The central smaller cell contains several empty vacuoles which may have been lamellar bodies, suggestive of a PN2. Some distended cristae in mitochondria. Z 15769: Again, several cuboidal epithelial cells, consistent with bronchiolar epithelium. The two central cells contain a few lamellar bodies and are probably PN2s. The smaller darker cell appears to be a basal cell. Z 15770: Alveolus with lining cells and underlying collapsed alveolar septa. The small dark lining cell contains four lamellar bodies and is a PN2. The other two lining cells are P N ls. Z 15771: Area seems to be a terminal bronchiole junctioning with an alveolar area with a flattened PN1 (lower right). Center top is a PN2 with five lamellar bodies. It appears to squeeze in between two cuboidal epithelial cells. At the right and left are capillaries. The right one contains a lamellar body in its lumen. Z 15772: Two large cuboidal epithelial cells or pneumocytes and a PN1 lining cell (bottom). Some dilated cristae in mitochondria noted. Conclusions: Appears normal. A few random PN2s with lamellar bodies noted. 000462 418-014:PAGE F-16 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014: EM -99.46 Animal No.: 18915-1___________ Treatment Group: 0 mg/kg. Control Sex: non applicable_________ Species/Strain: RCarlt:CD(BBR v a f/pius^ Tissue:_____ Liver ____________ Block No(s).: 99.46-7__________ Z15763 - Photo/Negative No(s).: Z 15767 Significant Lesions (check one): _________Yes X No Interpreting Pathologist (signature and date): __________________ Features: Z 15763: Hepatocyte in photograph has a large area o f cytoplasm filled with abundant glycogen-like material. Plasma membranes and endothelial cells are poorlydefined due to poor fixation. Mitochondria are dense and contain some dense deposits and dilated cristae. There is dilation of the RER, showing as round blebs on cross-section. A bile canaliculus is present in lower left. Fourteen (14) peroxisomes counted. Z 15764: Hepatocyte, similar to Z 15763. Five (5) peroxisomes counted. Z15765: Hepatocyte. Poor fixation clearly evidenced by loss of endothelial cells lining sinusoids. Eight (8) peroxisomes counted. Z 15766: Six (6 ) peroxisomes counted. Much of the cytoplasm contains fine dispersed granules, probably mostly glycogen, but also consistent with poor fixation. Z 15767: Hepatocyte similar to those described above. Many dilated blebs of RER. Eleven (11 ) peroxisomes counted. Conclusions: Suboptimal fixation of tissue. Number of peroxisomes counted: 8.8/cell (five counted). 000463 418-014:PAGE F-17 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014: EM -99.46 Animal No.: 18915-1____________ Treatment Group: 0 me/kg. Control Sex: non applicable_________ Species/Strain: RCarlt:CDBR VAF/Plus Tissue: Lung_________________ Block No(s).: 99.46-8__________ Z15758 - Photo/Negative No(s).: 157 62 _____ Significant Lesions (check one): _________Yes X No Interpreting Pathologist (signature and date): ___________________ Features: Z 15758: Collapsed alveolar septa. Air space at bottom is lined by a thin electron-dense cytoplasmic margin containing a few organelles and a large dilated bleb. Two capillaries in the collapsed area contain some lamellar structures free within the lumina. Lamellar bodies within pneumocytes or free in the air spaces are not evident. Z 15759: Alveolus and septa, mostly collapsed. PN Is line air spaces. Capillary at top left appears to contain a lamellar body within the endothelial cell cytoplasm. Smaller dark nucleus in the center appears to be a nucleus o f a PN1. Z 15760: Area o f collapsed septa, with air space to left. The space is lined by thin margin o f PN1. Subjacent to this space is a capillary containing a large lamellar body within the lumen. At top right is a cell, probably a PN2, which contains a single lamellar body in its cytoplasm. The clear cell in the center appears to be a pneumocyte. Z 15761: Photograph contains several capillaries in the collapsed septa. The large lumen at the bottom contains some pale-staining lipid material intermixed with endothelial cell cytoplasm. Air space is lined by flattened PN1 cytoplasm. N o lamellar bodies noted in pneumocytes. This profile in lumen may be tubular surfactant (lamellar body). Z 15762: Central dark cell appears to be a PN2 with a few lamellar bodies in its cytoplasm. Flattened PN1 cytoplasm lines the remainder of the air space. Tw o capillary lumina, dorsal to and to the right of this PN2, contain some lamellar material. A focus o f lamellar material is in the airway lumen._____________________________________________ Conclusions: Normal fetal lung. One PN2 was identified with some lamellar bodies. Other lamellar like structures were free within capillary lumina, and a single focus o f lamellar material was present in airway lumen.____________________________________ 000464 418-014PAGE F-18 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014: EM -99.46 Animal No.: 18915-2___________ Treatment Group: 0 me/kg. Control Sex: non applicable_________ Species/Strain: Crl:CDBR VAF/Piusro -Em Tissue: Liver ___________ Block No(s).: 99.46-9_____________ Z15753 - Photo/Negative Nofs).: Z 15757 Significant Lesions (check one): _________Yes X No Interpreting Pathologist (signature and date): ___________________ Features: Z 15753: Hepatocyte with numerous dilated blebs of RER present. Some dilatations noted in nuclear envelope. Abundant granular glycogen in cytoplasm. Mitochondria appear dense with some granular densities and some dilated cristae. Nine (9) peroxisomes counted. Z 15754: Hepatocyte surrounded by erythrocytes and hematopoietic cells. Endothelial cells are poorly defined due to poor fixation. Hepatocyte contains many dilated segments and blebs o f RER and nuclear envelope. Five (5) peroxisomes counted. Z 15755: Large central hepatocyte with abundant glycogen. Blebbing o f RER abundant. Two (2) peroxisomes counted. Z 15756: Field with several hepatocytes, however, only one at top center with nucleus is a complete cell. Cells has numerous dilated blebs o f RER, some dark and some light staining. Some dilated cristae noted in mitochondria. Four (4 ) peroxisomes counted. The cell at lower right appears to have more peroxisomes, but is an incomplete cell for counting. Z I5757: Several hepatocytes, although center cell with nucleus is only complete cell. It has dense mitochondria with granular densities and numerous dilated blebs o f RER. Twelve (12) peroxisomes counted. Conclusions: Average number of peroxisomes = 6.4 (5 cells counted). Suboptimal fixation. 000465 418-014PAGE F-19 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014: EM -99.46 Animal No.: 18915-2___________ Treatment Group: 0 me/kg. control Sex: non applicable_________ Species/Strain: Crl:CD(B)BR VAF/Plusc Jk Tissue: Lung _______________ Block No(s).: 99.46 -1 0 _________ Z15748 - Photo/Negative No(s).: Z 15752 Significant Lesions (check one): _________Yes X_____ No Interpreting Pathologist (signature and date): ____________________ Features: Z 15748: Two alveoli with intervening septum and capillary. P N ls line the septum, and there is a large cell attached to the right. This cell has minimal cytoplasmic organelles, might be a lymphocyte. Some lamellar material is present within the capillary lumen, but is not present in pneumocytes in this photograph. Z 15049: Alveolus, lining cells, and collapsed septa. Lining pneumocyte contains a single lamellar body in its cytoplasm just beneath the nucleus. At central top are several lamellar bodies with cytoplasm of probably PN2s. Z 15750: Cuboidal epithelium of probably bronchiole. Centrally is a PN2 with 5 lamellar bodies. Z 15751: Alveolus and lining PM Is. At lower right is a capillary with a lamellar body in the lumen of the capillary. At top left, in collapsed septal area, is a PM2 with several lamellar bodies. Z 15752: Alveolus on left with lining pneumocytes is free o f lamellar bodies. At top right is a single cell with three vacuoles which appear to have lamellar structures in them. Two capillaries at center and lower right. Conclusions: Several PN2 and lamellar bodies identified. Some lamellar bodies were in capillary lumina. 000466 418-014.PAGE F-20 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014: EM -99.46 Animal No.: 18952-1____________ Treatment Group: 1.6 mg/ke. high-dose Sex: non applicable_________ Species/Strain: Crl:CDBR VAF/Plus -Rat Tissue: Liver_________________ Block No(s).: 99.46-15_________ Z15722 - Photo/Negative N o (s ).: _ Z 1 5 7 2 6 _ _ Significant Lesions (check one): X Yes _________ No Interpreting Pathologist (signature and date): ___________________ Features: Z 15722: Hepatocyte. Cytoplasm contains abundant fine granular material, glycogen. Some dilated blebs of RER. Mitochondria appear dense with granular deposits and some dilated cristae. Twenty (20) peroxisomes identified. Adjacent endothelial cell is poorly-defined (poor fixation). Z 15723: Similar to Z 15722. Numerous round dilated blebs o f RER. Bile canaliculus present in bottom o f photograph. Ten (10 ) peroxisomes counted. Z 15724: Sinusoid containing several erythrocytes. Segment o f hematopoietic granulocytic cell in top left. Endothelial cell and lipid-containing perisinusoidal cell are poorly fixed. Two incomplete hepatocytes on right. A few peroxisomes are noted, but not counted. Z 15725: Hepatocyte. Bile canaliculus at top; sinusoid in lower right of photograph. Numerous dilated blebs o f RER. Fifteen (1 5 ) peroxisomes counted. Z 15726: Hepatocyte. Bile canaliculi at top right and left. Numerous, variably-sized dilated blebs of RER. Nineteen (19) peroxisomes counted. Conclusions: Increased numbers of peroxisomes. Average 16 peroxisomes/hepatocyte (4 cells counted). Suboptimal fixation. 000467 418-014.PAGE F-21 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014: EM -99.46 Animal No.: 18952-1___________ Treatment Group: 1.6 me/kg. high-dose Sex: non applicable_________ Species/Strain: Crl:CDBR VAF/Plus Jk Tissue: Lung _______________ Block No(s).: 99.46-16________ Z15717- Photo/Negative N o (s ).: _ Z 1 5 7 2 1 _ _ Significant Lesions (check one): X Yes _________ No Interpreting Pathologist (signature and date): ___________________ Features: 15717: Alveolus with lining cells. Two large P N ls are present and appear to have a PN2 between them. However, a PN1 margin still separates the PN 2 from the sinusoid. Several lamellar bodies are present associated with the PN2, and some o f them appear to be extracellular. The PN1 on the left contains abundant fine granular material in its cytoplasm, consistent with glycogen. Z 15718: Collapsed area of alveolar septa. Two central cells contain lamellar bodies, three (3) in one, and seven (7) in the other. Both cells appear to be PN2s, although normal pulmonary architecture is unclear because o f the collapsed state. Z 15719: Two cells with abundant centrioles appear to represent bronchiolar cells, and the centrioles are associated with cilia. Centrally are two cuboidal cells. One contains multiple (5 ) lamellar bodies. This cell appears to be a PN2. A large lamellar body is associated with the left cell, but it may be extracellular. Both cells contain abundant granular material, consistent with glycogen. Z 15720: Alveolus and lining cells. Two cells contain lamellar bodies, six in one and eight in the other. Some lamellar material appears to be extracellular. Z 15721: A v eolu s, five lining cells, and a capillary in top right o f photograph. This is possibly a junction with a terminal bronchiole. Two of the pneumocytes contain multiple lamellar bodies, and appear to be PN2s. Conclusions: There appear to be increased numbers o f PN2s containing lamellar bodies. Some extracellular lamellar material noted. 000468 418-014:PAGE F-22 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _ 41 8-014; EM -99.46 Animal No.: 18952-2_____________ Treatment Group: 1.6 mg/kg. high-dose Sex: non applicable_________ Species/Strain: Crl:CDBR VAF/Plus -Em Tissue: Liver________________ Block No(s).: 99.46-17________ Z15712- Photo/Negative No(s).: Z I5716 Significant Lesions (check one): X Yes _________ No Interpreting Pathologist (signature and date): __________________ Features: Z 15712: Hepatocyte. Poorly-defined sinusoids and endothelium at top right and bottom of photograph. The hepatocyte cytoplasm appears voluminous with abundant fine granular material consistent with glycogen. Mitochondria appear dense and have some granular deposits and dilated cristae. Some dilated blebs o f RER. Eight (8) peroxisomes identified. Z 15713: Hepatocyte. Hematopoietic cell to right. Bile canaliculus present in lower right. Abundant fine granular material, glycogen, in cytoplasm. Appear to be increased numbers o f peroxisomes. Twenty-seven (2 7 ) peroxisomes present. Z 15714: Hepatocyte. Many dilated blebs o f RER and abundant glycogen particles in cytoplasm. Fourteen (14) peroxisomes identified in this cell. Z 15715: Hepatocyte. Many dilated blebs of RER and abundant glycogen particles in cytoplasm. Mitochondria with some granular deposits and dilated cristae. Nineteen (19) peroxisomes identified in this cell. Z 15716: Sinusoid with lining endothelial cell. Cellular fixation is poor. Adjacent hepatocytes contain many mitochondria, RER and glycogen-like particles. Dilated blebs o f RER are present. Multiple peroxisomes are present, but not counted because whole hepatocytes are not in photograph. Conclusions: Hepatocytes appear to contain increased numbers o f peroxisomes, average 17/hepatocyte (4 cells counted). Cells contain abundant glycogen. Suboptimal cellular fixation. 000469 418-014:PAGE F-23 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014: EM -99.46 Animai No.: 18952-2___________ Treatment Group: 1.6 rne/ke. high-dose Sex: non apolicable_________ Species/Strain: RCarht CDOBR v a f/pius(3 Tissue: Lune__________________ Block No(s).: 9 9.46 -1 8 ________ ZI5707 - Photo/Negative No(s).: Z 15711 Significant Lesions (check one): X Yes _________ No Interpreting Pathologist (signature and date): ___________________ Features: 15707: Single PN2 lining alveolus. Cell contains multiple mitochondria around the nucleus, and also multiple clear vacuoles, which are mostly empty of contents. These were probably lamellar bodies which were extruded into the alveolar space. These vacuoles are lined by a single membrane, whereas the mitochondria have a double membrane. 15708: Collapsed alveolar septa. A thin layered P N 1 covers the surface. Beneath is an endothelial cell with many pinocytotic vesicles, one a single lamellar body. Centrally is an area o f flattened basement membrane material containing many collagen fibrils. A dark cell is present in the upper right, which contains many mitochondria. To the left of this cell are two lamellar bodies, although they appear to be in a cytoplasmic extension of an unidentified cell. Z15709: A nucleated PN2 containing multiple lamellar bodies (5) appears to be overlain by a PN1 lining the alveolus. Z15710: Alveolar space in top right is lined by a thin PN1 cell. Adjacent to this PN1 and covered by it's cytoplasm is an extracellular area with several (3) lamellar bodies. A capillary is dorsal containing an erythrocyte and endothelial cell. Several cells within collapsed septa are not clearly identifiable as to cell type. Z15711: A single dark PN2 is squeezed between two PN ls. Its cytoplasm contains several empty vacuoles, which have apparently extruded lamellar bodies. To the right of this cell are two large lamellar bodies within some lipid-like material. These bodies may be in the adjacent PN1, macrophage, or may be extracellular. A small capillary is in the lower right margin of the photograph.____________________________________________________________________________ Conclusions: There appear to be increased numbers o f type II pneumocytes (PN 2s), and more lamellar bodies were seen in this sample. Otherwise, cellular morphology appeared essentially normal. Fixation appeared satisfactory. 000470 418-014:PAGE F-24 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: _ 41 8-0,14-uEM -99.46___ Animai No.: 18956-1__________________ Treatment Group: 1.6 me/kg. high-dose Sex: non applicatale_________ Species/Strain: Crl:CD<S>BR v a f/piu. J&ai Tissue: Liver ______________ Block No(s).: 99.46-13________ Z15732 Photo/Negative No(s).: Z 15736 Significant Lesions (check one): X Yes _________ No Interpreting Pathologist (signature and date): ___________________ Features: Z 15732: Hepatic sinusoid containing nucleated erythroycte and a couple mature erythrocytes. Some lamellar debris is present in the sinusoid. Two endothelial cell nuclei are present. There are no complete hepatocytes in this photograph. Z 15733: Hepatocyte contains abundant granular particles consistent with glycogen, and numerous RER palisades. Mitochondria appear dense and have some granular deposits and dilated cristae. Eleven (11) peroxisomes counted. A bile canaliculus is present between two hepatocytes. Z 15734: Hepatocyte. Similar to Z 15733. Numerous dilated blebs of RER are present. Twenty-four (24) peroxisomes. Z 15735: Hepatocyte with similar morphology as described for above cells. Sixteen (16) peroxisomes counted. Z 15736: Hepatocyte. Dilated blebs o f RER present Mitochondria are dense with granular deposits and dilated cristae. Eighteen (18 ) peroxisomes. Conclusions: Increased numbers o f peroxisomes per hepatocyte. 17.25 peroxisomes per hepatocyte (average o f four counted cells). Suboptimal fixation. 000471 418-014.PAGE F-25 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014: EM -99.46 Animai No.: 18956-1___________ Treatment Group: 1.6 me/ke. high-dose Sex: non ap p lica le_________ Species/Strain: Crl:CDBR Rat VAF/Plus T is s u e : _______Lune_______________ Block No(s).: 99.46-14________ Z 15727- Photo/Negative No(s).: Z 15731 Significant Lesions (check one): X Yes _________ No Interpreting Pathologist (signature and date): ___________________ Features: 15727: Alveolus with adjacent collapsed septa. The two bottom pneumocytes are PN2 with lamellar bodies. Some of the bodies are partially empty, and in one cell appear to be extruding material extracellularly with other lipid material. At the top is a large PN1. At the left is a capillary with a nucleated erythrocyte. Z 15728: Collapsed alveolar septa. Several cells contain lamellar bodies. At left one lamellar body appears to be within the lumen of a capillary. Z 15729: Somewhat T-shaped capillary containing an electron-dense erythrocyte. The endothelial cell nucleus is ventral to the erythrocyte. Within the lumen of the capillary are several lamellar bodies. At left o f field is the thin cytoplasmic lining o f a PN1. The other cells appear to be sections through pneumocytes at various angles. Z 15730: Alveolar lumen and lining cells. Centrally is a small PN2 containing a couple small lamellar bodies next to its nucleus. The dark-staining cytoplasm to the right o f this cell appears to be an extension from an alveolar macrophage. To the left are a couple pneumocytes, and they contain a couple lamellar bodies. Z 15731: Alveolar lumen and lining cells. Top left cell is a PN2 with two lamellar bodies. A right central, pale-staining, pneumocyte also contains several small lamellar bodies. The septa are collapsed in this area. Conclusions: Increased number o f pneumocytes containing lamellar bodies. Some lamellar bodies noted in capillaries. 000472 418-014-.PAGE F-26 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014: EM -99.46 Animal No.: 18973-1___________ Treatment Group: 1.6 me/kg. high-dose Sex: non applicable_________ Species/Strain: CRarlt:CDBR VAF/Plusfl) Tissue: Liver________________ Block No(s).: 99.46-11________ Z15742 - Photo/Negative Nois).: Z 15747 Significant Lesions (check one): X Yes _________ No Interpreting Pathologist (signature and date): ___________________ Features: Z 15742: Hepatocyte contains abundant fine granular material consistent with glycogen in the cytoplasm. There are numerous dilated blebs o f RER on cross section. Mitochondria appear dense and have some granular deposits. Seventeen (17) peroxisomes counted. Some lamellar material noted extracellularly and possibly within clear spaces in the left portion of the hepatocyte. Z 15743: Hepatocyte, overall similar to Z 15742, although fixation appears better. Nine (9) peroxisomes counted. Lamellar material at top appears to be extracellular. Z 15744: Hepatocyte. At left is a hematopoietic cell in sinusoid and at upper right appears to be a dilated bile canaliculus. Abundant area o f glycogen in cytoplasm. Nineteen (19) peroxisomes counted. Z 15745: Sinusoid, endothelial cell and incomplete portions o f a couple hepatocytes. Centrally is a clear cytoplasmic area with many dilated blebs o f RER. A few peroxisomes are present. No peroxisome count made. Z1546: Hepatocyte contains abundant fine granular material consistent with glycogen in the cytoplasm. There are numerous dilated blebs o f RER on cross section. Mitochondria appear dense and have some granular deposits and some dilated cristae. Eleven ( 11) peroxisomes counted. Z 15747: No photograph. Negative was too dark. With five photographs already evaluated, it is not necessary to retake or try to print this image.________________________ Conclusions: Appear to be increased peroxisomes per hepatocyte. Average 14/cell(four cells counted. Suboptimal fixation. 000473 418-014:PAGE F-27 TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION Study No.: 418-014: EM -99.46 Animai No.: 18973-1___________ Treatment Group: 1.6 me/kg. high-dose Sex: non applicatile_________ Species/S train: RCralt:CD<S)BR v a f/pius<R) Tissue: L u n e ________________ Block No(s).: 99.46-12 Z15737 - Photo/Negative No(sl.: Z 15741 Significant Lesions (check one): X Yes _________ No Interpreting Pathologist (signature and date): ___________________ Features: Z 15737: Collapsed alveolar septa. Centrally is a PN2 with approximately eight lamellar bodies, of which several appear to be releasing their contents into the extracellular space. Ventrally are also segments o f a cell's cytoplasm that appears to contain some lamellar body vacuoles. A couple P N ls are present adjacent to the PN2, and a capillary with erythrocyte and endothelial cell is present in lower left central area o f photograph. Z 15738: Alveolus with lining cells. The large nucleus belongs to a PN1 whose cytoplasm lines the air space. Beneath this cell and just to the right are several lamellar bodies, although it's in a subjacent capillary. To the middle left is a capillary with an eyrthrocyte that does contain some lamellar debris in the lumen. Z 15739: Alveolus and collapsed septa. Similar to Z 15738, some lamellar bodies appear to be within the lumen o f a capillary. P N ls line the alveolar space. Z 15740: Large capillary with several erythrocytes to the right. Left of this is a cellular extension, probably a PN2, that contains a distinct lamellar body. A thin layer o f PN1 lines the alveolar space. Z 15741: Alveolar space contains abundant lamellar material. Several P N ls are present along the lining, but no PN2 are present. Lamellar bodies are present in one capillary lumen and in some interstitial spaces in the right side o f the photograph.________________ Conclusions: Appear to be increased lamellar bodies in the tissue compared with control sample, although many were in capillary Iumina and some in alveolar and interstitial spaces. 000474 418-014:PAGE F-28 IH. Transmission Electron Micrographs 000475 418-014:PAGE F-29 NOTE: The original glossy figure photographs (Figures 1 - 1 2 ) were delivered to Dr. Marvin T. Case with the draft pathology report on March 19, 1999. 000476 418-014:PAGE F-30 PrimedicAa AncriglluasryStPuadtyhoNloog.y41R8e-p0o1r4t Figure Legends Figure 1. Liver. Hepatocyte showing normal cytoplasmic organelles. White spots within mitochondria represent some dilatation o f cristae. Some peroxisomes are identified. To the right is an endothelial cell and erythrocytes of adjacent sinusoid. Animal No. 189041, 0 mg/kg/day (control), 1 1.200X magnification. Figure 2. Liver. Hepatocyte. Dilatation o f nuclear envelope and rough endoplasmic reticulum (blebbing) indicates suboptimal fixation. Mitochondria are dense and show some granular deposits and dilated cristae. Some representative peroxisomes are labeled. Hematopoietic cells are present in sinusoid at upper right o f photograph. Animal No. 1 8 9 1 2 - 1 ,0 mg/kg/day (control), 1 1,200X magnification. Figure 3. Liver. Cellular morphology is similar to Figure 2. Some peroxisomes are labeled, and appear more numerous than in controls. A bile canaliculus is present in upper right o f photograph and hematopoietic cells are present in sinusoid at upper left. Animal No. 18952-2, 1.6 mg/kg/day (high-dose), 1 l,2 0 0 X magnification. Figure 4. Liver. Dilatation of rough endoplasmic reticulum is evident. Some peroxisomes, increased in number, are labeled. Animal No. 18956-1, 1.6 mg/kg/day (high-dose), 11,2 0 0 X magnification. Figure 5. Liver. Hepatocyte demonstrating somewhat better fixation with more even dispersion o f organelles. Clear space at top o f cell, which contains some membranous debris, appears to be extracellular in the perisinusoidal space. Animal No. 18973-1, 1.6 mg/kg/day (high-dose), 11,200X magnification. Figure 6. Lung. Type II pneumocyte is present along the alveolar wall, adjacent to type I pneumocyte. Lamellar bodies within the cytoplasm identify this cells as a type II pneumocyte. Animal No. 18904-1, 0 mg/kg/day (control), 15.680X magnification. Figure 7. Lung. Three cuboidal pneumocytes line this airway space. A flattened type I pneumocyte is evident along the ventral portion o f the airway space. Two small lamellar bodies in one cuboidal cell suggest this is a type II pneumocyte which has secreted most o f its lamellar bodies and is destined to become a type I pneumocyte. Animal No. 189042, 0 mg/kg/day (control), 1 1.200X magnification. Figure 8. Lung. A more densely-staining type II pneumocyte, containing two lamellar bodies, appears squeezed between two type I pneumocytes. Some lamellar material is also present within the capillary lumen o f the septal wall both above and to the right of the type II pneumocyte. Animal No. 18915-1, 0 mg/kg/day (control), 1 1,200X magnification. 000477 418-014:PAGE F-31 PrimedicaAAncriglluasryStPuadtyhoNloog. y41R8e-p0o1n4 Figure 9. Lung. Dense-staining cell is a type II pneumocyte positioned between two type I pneumocytes. Four distinct lamellar bodies are present in the cytoplasm o f the type H pneumocyte, although the lamellar material is partially washed out in these vacuoles. To the right o f this cell are two dark lamellar-type bodies surrounded by some lipid. This appears to be within a cell in a collapsed area o f septa, and may be a macrophage. Animal No. 18952-2, 1.6 mg/kg/day (high-dose), 11.200X magnification. Figure 10. Lung. Collapsed area o f septa showing two type II pneumocytes containing lamellar bodies. A type I pneumocyte is to the left of these cells. Capillaries and adjacent alveolar cells are present dorsally and to the right o f the photograph. Animal No. 189521, 1.6 mg/kg/day (high-dose), 11,200X magnification. Figure 11. Lung. Centrally is a cell containing several lamellar-like structures which appears to be a type II pneumocyte. Glycogen-like material appears to fill much o f the cytoplasm. Adjacent cells to the right contain centrioles and represent ciliated bronchiolar cells. Cuboidal epithelial cells are to the left and ventral to the type II pneumocyte. Animal No. 18952-1, 1.6 mg/kg/day (high-dose), 11,200X magnification. Figure 12. Lung. Alveolar space contains abundant lamellar-like material, which is consistent with surfactant secreted from type II pneumocytes. A type I pneumocyte lines the space on the left and right. Two cuboidal cells containing glycogen-like granular material appear to be pneumocytes, probably type Es which have released their lamellar bodies. Animal No. 18973-1, 1.6 mg/kg/day (high-dose), 11,200X magnification. 000478 Abbreviation A BC BE CL Cap CN CP D DD DC DN DR EC ER G HC L LB M N P PN1 PN2 PM RER 418-014:PAGE F-32 PrimedicaAAncriglluasryStPuadtyhoNloog. y41R8e-p0o1r4t Index of Labels for Ultrastructural Structures Structure Alveolar space Bile canaliculus Bronchiolar epithelium Capillary lumen Capillary Centrioles Cuboidal pneumocyte Desmosome Dense deposits Dilated cristae Dilated nuclear envelope Dilated RER Endothelial cell Erythrocyte Glycogen Hematopoietic cells Lipid droplet Lamellar body Mitochondria Nucleus Peroxisome Type I pneumocyte Type II pneumocyte Plasma membrane Rough endoplasmic reticulum 000479 BEST COPY AVAILABLE 418-014:PAGE F-35 A i} - J C 0 0 0 4 8 ,*> T. 418-014.PAGE F-45 IV. Quality Assurance Statement 000492 418-014:PAGE F-46 Pathology Associates international A Company of Science Applications International Corporation QUALITYASSURANCE STATEMENT This electronmicroscopystudyhas beeninspectedandauditedbytheQuality AssuranceUnitas requiredbytheGoodLaboratoryPracticesRegulationspromulgated bytheU.S. FoodandDrugAdministration. PAIhasafunctioningandresponsive QualityAssuranceUnitwhichreportsdirectlytomanagement. Thefollowingisarecord ofinspections/auditsandtheirresultingreports: Date of Inspection PhaseInspected DateFindings Reported toManagement andStudyDirector 2/18,19/99 3/16/99 3/17/99 4/5/99 ProcessingandEmbedding DataAudit Draft ReportAudit Final ReportAudit 2/19/99 3/17/99 3/17/99 4/5/99 Sponsor: 3MCorporateToxicology Test Article: PFOS StudyNumber. 418-014, PAIStudyEM-99.46 000493 4915DProspectusDrive Durham, NorthCarolina 27713(919) 544-5257(919) 544-3218FAX APPENDIX G STATEMENT OF THE STUDY DIRECTOR 000494 418-014.PAGE G-1 Arg9u0s5TReSelhTseeepeealhehrocHfynahoxeDLr::sra((hi2vb2a1eo1m,5r5a,)B)t4Po4u4A4riil33ed--1si88n,957g0I18n4Ac074. PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS SPONSOR'S STUDY NUMBER: T-6295.13 STATEMENT OF THE STUDY DIRECTOR This final report accurately reflects the raw data obtained during the performance of the study. No deviations from the U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations; Final Rule3, the Japanese Ministry of Health and Welfare (MHW) Good Laboratory Practice Standard for Safety Studies on Drugsband the European Economic Community (EEC) Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles of good laboratory practicecoccurred that affected the quality or integrity of the study. 23L-E&ll - 99 Raymond G. York, RtUD., DABT Associate Director of Research and Study Director a. U.S. Food and Drug Administration. Good Laboratory Practice Regulations; Final Rule. 21 CFR Part 58. b. Japanese Ministry of 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 decision/recommendation on compliance with principles of good laboratory practice. Official Journal of the European Communities: Legislation. 32(No. L 315; 28 October): 1-17. 000495 APPENDIX H QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT 000496 OPrimedica 418-014.PAGE H-1 Aig9u0s5TReSelThseeepeaelherhoHcfyanhoxeDrL::sra((hi22bva1oe1m,5r5a,)B)t4Po4u4A4rii3l3ed--1si88n,957g0In814Ac074. QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT Study Director: Raymond G. York, Ph.D., DABT Executive Director of Research: Mildred S. Christian, Ph.D., Fellow, ATS Protocol 418-014: Oral (Gavage) Cross-Fostering Study of PFOS in Rats Sponsor's Study Number: T-6295.13 The draft protocol for this study was audited for adherence to U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations, Japanese Ministry of Health and Welfare (MHW); Good Laboratory Practice Standard for Safety Studies on Drugs, and European Economic Community (1989) council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles of good laboratory practice on 19 OCT 98. Critical phases of this study were inspected nine times; study information and raw data were audited twice (see tables 1 and 2 for dates and phases/data). The draft final report and the raw data for this study were compared and audited for accuracy, for adherence to protocol requirements, and for adherence to U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations, Japanese Ministry of Health and Welfare (MHW); Good Laboratory Practice Standard for Safety Studies on Drugs, and European Economic Community (1989) council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles of good laboratory practice between 21 MAY 99 and 10 JUN 99, and for revisions requested by the Sponsor on 20 JUL 99, 22 JUL 99, and for finalization on 23 JUL 99. 000497 418-014:PAGE H-2 This study was conducted according to U.S. Food and Drug Administration (FDA) Good Laboratory Practice Regulations, Japanese Ministry of Health and Welfare (MHW); Good Laboratory Practice Standard for Safety Studies on Drugs, and European Economic Community (1989) council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decision/recommendation on compliance with principles of good laboratory practice. Nancy J. Gongliewski Date Quality Assurance Manager Quality Assurance Supervisor and Principal Auditor 000498 TABLE 1 CRITICAL PHASES INSPECTED 418-014.PAGE H-3 Test Article Administration - Gavaae Date of inspection: 03 NOV 98 Date results reported to the Study Director and Management: 06 NOV 98 Test Article Preparation Date of inspection: 03 NOV 98 Date results reported to the Study Director and Management: 06 NOV 98 Cohabitation Date of inspection: 17 DEC 98 Date results reported to the Study Director and Management: 18 DEC 98 Natural Deliverv/Litter Observations Date of inspection: 06 JAN 99 Date results reported to the Study Director and Management: 19 JAN 99 Blood Collection - Dams and Puds Dates of inspection: 28 JAN 99, 28 JAN 99 Dates results reported to the Study Director and Management: 02 FEB 99, 02 FEB 99 000499 418-014:PAGE H-4 Fecal and Urine Collection Date of inspection: 28 JAN 99 Date results reported to the Study Director and Management: 02 FEB 99 Scheduled Sacrifice - Dams and Puds Dates of inspection: 28 JAN 99, 28 JAN 99 Dates results reported to the Study Director and Management: 02 FEB 99, 02 FEB 99 000500 418-014:PAGE H-5 TABLE 2 RAW DATA AUDIT(S) The following study information and raw data were audited from 26 FEB 99 to 10 MAR 99: Animal receipt, randomization, physical examination and acclimation. In-life transaction record. Feed consumption. Cohabitation. Natural delivery observations. Litter observations. Pup body weights and status. Table of random units. Milk collection data and packing lists. Blood and liver collection data and packing lists. Urine and fecal collection data and packing lists. Necropsy. Tissue packing lists. Male breeder colony records. General comments. Study maintenance records. Temperature and relative humidity reports. Feed, water and bedding analyses. Edit requests. Dosage volumes. Data review pages. The results of this audit were reported to the Study Director and Management on 12 MAR 99. The following study information and raw data were audited from 26 FEB 99 to 10 MAR 99: Vehicle receipt, preparation and use. Test article receipt, preparation and use. Test article packing lists. The results of this audit were reported to the Study Director and Management on 12 MAR 99. 000501