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FINAL REPORT 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
ocasss
PROTOCOL 418-014
ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS
SPONSOR'S STUDY NUMBER: T-6295.13
TABLE OF CONTENTS SUBJECT 1. SUMMARY AND CONCLUSION A. Methods B. Results C. Conclusion II. DESCRIPTION OF TEST PROCEDURES A. Conduct 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 il-1 11-1 11-1 11-1 11-1 11-1 11-1 11-1 11-1
i 001288
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
ii
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 N-4 II-4
001287
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
in
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
001288
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 Necropsy Observations 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 II-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 HI-1 111-1
III-2 III-2 III-2 III-2
0012S9
SUBJECT
E. Clinical Observations from Birth to Day 21 Postpartum and Necropsy Observations
PAGE III-3
F. Electron Microscopic Evaluation of theLiver 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 00129
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-45
Table 23.
Maternal Body Weights - Lactation - Individual Data Fo Generation Female Rats
B-53
001291.
vi
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
VII
G-1
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SUBJECT
APPENDIX H - QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT
PAGE H-1 to H-5
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viii
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
l- SUMMARY AND CONCLUSION
A. Methods3
Seventy-eight virgin female Cri: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
1 1 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
Reassigned 13
12 12 13
R eassigned Litter Type
(Dam Treatm ent) Treated
Untreated Treated
Untreated
i 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).
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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
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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 P ups 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.
001296
B. Results
418-014:PAGE I-4
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
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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.
001298
C. Conclusion
418-014:PAGE I-6
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
Z3-TUI-) ?
Date
Study Director
Z Z ^L L = 3 L l Date
001799
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 CrliCDOBR 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
001300
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
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A.16. Dates of Technical Performance:
Rat Arrival
27 OCT 98
Dosage Period (42 days before cohabitation
until DLa21)
02 NOV 98 -2 8 JAN 99
Cohabitation Period
13 DEC 98 PM - 19 DEC 98 AM
Day 0 of Presumed Gestation (DG 0)
14 DEC 98 - 19 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 99 - 22 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.
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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.
001C03
418-014:PAGE II-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* 1g
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).
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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 fo r 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 Facility<5'7); 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 o f 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 computergenerated (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
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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
1 1 II II
Number of M ated 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 Treatm ent) Treated
U n tre ate d
Treated
U n tre ate d
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.
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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 of ten 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 Animalsw.
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.
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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.
001308
G. Methods: G.1. Dosage Administration:
418-014:PAGE 11-10
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
Utters
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).
003O3
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:
4
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.
001310
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 1, 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
001311
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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 livebom 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.
001312
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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
001313
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.
001314
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.
001315
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.
001316
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),
001317
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/dav) 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
001318
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 Day 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.
001319
418-014;PAGE MI-4 F. Electron Microscopic Evaluation of Liver and Lung from Day 1
Postpartum P uds (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.
001320
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, Vol. 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.
001321
APPENDIX A REPORT FIGURES
001322
MATERNAL BODY WEIGHTS
Figure 1
450
0 (VEHICLE) MG/KG/DAY
1 8 MG/KG/DAY
418-014:PAGE A-1
001323
DAY OF STUDY
DAY OF GESTATION
i
MATERNAL BODY WEIGHTS
RATS ASSIGNED TO CROSS-FOSTERING Figure 2
SUBSET A a
SUBSET B b
-~o~
SUBSETCc
--A--
SUBSET D d
418-014:PAGE A-2
001324
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 liners. 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.
APPENDIX B REPORT TABLES
001325
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 1 (PAGE 1) : CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS
DOSAGE GROUP 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
INCISORS: TOTAL MISALIGNED MISSING/BROKEN
104/ 49/ 57/ 32/
11/
5 3 2 1 I
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 I. b. Located on the head, neck, tail or left forelimb.
II 1.6
1512/ 36
0
3/ 2
35/ 1
1/ 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
001326
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
CD
o\
MAXIMUM POSSIBLE INCIDENCE
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
IS/ 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
152/ 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
001327
418-014:PAGE B-3
001328
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
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 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.
0/ 0
D II d 1.6
267/ 13 0
22/ 1 22/ 1
7/ 1 3/ 1 0/ 0 0/ 0 0/ 0 0/ 0
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 2 (PAGE 1): NECROPSY OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS
DOSAGE GROUP DOSAGE (MG/KG/DAY)
I 0 (VEHICLE)
II i.e
RATS EXAMINED a
N
41b 36
MORTALITY
N
00
APPEARED NORMAL
N
40 36
LEFT INGUINAL AREA: MASS
N
10
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
2CT00
(0
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
MEAN+S.D
231.4 + 8.6
DAY 8
MEAN+S.D
246.5 + 10.0
DAY 15
MEAN+S.D
257.9 + 11.5
DAY 22
MEAN+S.D
266.3 +_ 14.2
DAY 29
MEAN+S.D
274.7 14.7
DAY 36
MEAN+S.D
285.7 + 16.5
DAY 43a
MEAN+S.D
283.9 + 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 f 9.2 246.8 f 10.5 257.9 + 14.1 265.4 + 11.9 270.5 + 12.7 278.3 + 14.7 274.4 15.9
418-014:PAGE B-5
GCCTOO
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 4 (PAGE 1) : BODY WEIGHT CHANGES - PRECOHABITATION - SUMMARY - Fo GENERATION FEMALE RATS
DOSAGE GROUP DOSAGE (MG/KG/DAY)
I 0 (VEHICLE)
RATS TESTED
N
42
BODY WEIGHT CHANGE (G)
DAYS 1 - a
MEAN+S.D .
+15.1 7.0
DAYS a - 15
MEAN+S.D .
+11.4 + 6.3
DAYS 15 - 22
MEAN+S.D .
+8.4 f 7.7
DAYS 22 - 29
MEAN+S.D .
+8.3 + 4.8
DAYS 29 - 36
MEAN +S .D .
+11.0 6.1
DAYS 36 - 43a DAYS 1 - 43a
MEAN+S.D . MEAN+S.D .
-2.6 1 1 1 ( 17lb
+53.0 11.4
[ 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 8.2 +11.0 7.5
+7.5 + 4.9 +5.1 + 5.9 +7.8 5.6 -3.9 + 6.2 +43.2 4_ 14.3
418-014:PAGE B-6
0013
C4
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 5 (PAGE 1) : MATERNAL BODY HEIGHTS - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS
DOSAGE GROUP DOSAGE (MG/KG/DAY)
RATS TESTED
N
PREGNANT
N
MATERNAL BODY WEIGHT (G)
DAY 0
MEAN+S.D .
DAY 7
MEAN+S.D .
DAY 14
MEAN+S.D .
DAY 20
MEAN+S.D .
DAY = DAY OF GESTATION
I 0 (VEHICLE)
42 35
295.1 +16.9 326.8 + 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
418-014:PAGE B-7
001332
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
MEAN+S.D .
DAYS 7 - 14
MEAN+S.D .
DAYS 14 - 20
MEAN+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
418-014:PAGE B-8
001333
418-014:PAGE B-9
001334
PROTOCOL 418-014: ORAL (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
MEAN+S.D.
334.B + 20.1
331.8 + 29.9
326.6 + 20.0
DAY 4 DAY 7 DAY 10 DAY 14 DAY 22
MEAN+S.D. MEAN+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 11] f
334.2+ 18.2
l H]f 347.6 + 16.8
( Ulf 350.7 + 18.3
[ ll]f 354.7 + 19.1
DAY = DAY OF LACTATION ( ) = NUMBER OP 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.
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
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.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
MATERNAL BODY WEIGHT CHANGE (G)
12
Ilf
DAYS 1 - 4
MEANtS .D .
+0.9 + 14.8
+14.9 + 12.4
+3.0 + 7.3
DAYS 4 - 7
MEAN+S.D .
+11.9 + 10.7
+12.9 + 13.0
+8.1 + 6.7
DAYS 7 - 10
MEAN+S.D .
+8.6 + IS.3
+6.2 + 8.6
+13.4 + 9.0
DAYS 10 - 14
MEAN +S .D .
i +12.6 + 16.6
+2.1 + 13.7
+3.1 + 9.1
DAYS 14 - 22 DAYS 1 - 22
MEAN+S.D . MEAN+S.D .
- -6.9 + 13.7 +27.2 +15.1
-1.5 + 25.2 +34.7 + 17.5
+4.0 + 16.7 +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 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 pupa 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
I
418-014:PAGE B-10
001335
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER-, T-629S.13) TABLE 9 (PAGE 1): ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - PRECOHABITATION - SUMMARY - Fo GENERATION FEMALE RATS
DOSAGE GROUP DOSAGE (MG/KG/DAY)
RATS TESTED
u
I 0 (VEHICLE)
42
II 1.6
36
PEED 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.
H
19.5 + 1.7 I 40] a
20.1 + 1.7 ( 41] a
19.7 + 1.8 [ 41] a
19.6 + 2.2 l 40] a
19.6 + 2.1 ( 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 l 34] a
18.7 1.3 [ 34] a
18.3 + 1.6 l 34Ja
17.2 + 1.5 I 33] a
18.7 + 1.3 ( 33] a
DAYS - DAYS OF STUDY [ ] = NUMBER OF VALUES AVERAGED a. Excludes values that were incorrepfly 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
001336
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-POSTERING 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 X .6
RATS TESTED
N
42 36
FEED CONSUMPTION (G/KG/DAY)
DAYS 1 - 8 DAYS 8 - 15 DAYS IS - 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.
81.6 + 6.1 ( 40)a
79.7 + 5.4 l 41) a
75.0 + 5.1 l 41] a
72.7 + 6.9 ( 40)a
70.0 + 5.8 [ 40] a
63.8 + 7.7 l 15]a,c
74.2 + 4.9 [ 151a,c
82.4 + 8.1 l 34 ]a
78.0 + 5.2
72.1 + 4.6 I 34] a
69.7 + 3.9 [ 34 ]a
66.8 + 4.6 ( 34 ]a
61.9 + 3.9 ( 33] a
71.7 + 3.8 [ 331a
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-12
001337
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
MEAN+S.D .
DAYS 14 - 20
MEANtS.D .
DAYS 0 - 20
MEAN+S.D.
DAYS = DAYS OF GESTATION [ ] - NUMBER OP VALUES AVERAGED a. Excludes values Chat were associated with spillage.
I 0 (VEHICLE)
42 35
24.2 + 2.7 l 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
001338
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
MEAN+S.D .
DAYS 14 - 20
MEAN+S.D .
DAYS 0 - 20
MEAN+S.D.
DAYS = DAYS OF GESTATION [ ) = NUMBER OP VALUES AVERAGED a. Excludes values were associated with spillage.
I 0 (VEHICLE)
42
35
II 1.6
36
27
76.9 + 5.8 1 33) a
76.7 + 7.8
62.8 + 4.6
72.0 + 4.1
71.2 + 4.2
73.8 + 4.7
67.6 6.3
70.7 + 3.4
--
418-014:PAGE B-14
2 T00
CO
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-POSTERING 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)
A Ia 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/DAY)
DAYS 1 - 4
MEAN+S.D .
19.1 + 4.0
24.9 + 3.6
18.8 + 3.3
21.3 + 3.9
DAYS 4 - 7
MEAN+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
MEAN+S.D. MEAN+S.D. MEAN+S.D.
45.0 + 3.3 l 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
52.0 + 5.8 t I0)g
38.0 + 4.4 [ 10] g
43.3 + 3.3 53.3 + 5.1 40.2 + 3.7
DAYS = DAYS OF LACTATION l J = 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 iB presumed that the pups begin to consume maternal feed after day 14 of lactation, maternal feed consumption values
were not tabulated on days 14 to 21 of lactation.
418-014:PAGE B-15
001340
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 Ia 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
MEAN+S.D.
57.5 + 12.5
73.8 + 11.6
58.4 + 11.1
67.2 + 12.2
DAYS 4 - 7
MEAN+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 [ 10) g
159.7 + 12.0
118.8 + 8.9
141.9 + 14.0 [ nig
159.9 + 10.8
128.5 + 11.4
121.2 + 12.9
150.7 + 15.8 I 10 Jg
113.8 +12.6 ( 10] g
129.8 + 8.3 156.8 + 10.1 121.9 + 7.3
DAYS = DAYS OF LACTATION
( 1 = 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.
i
h. Because it is presumed that the pups begin to consume maternal feed after day 14 of lactation, maternal feed consumption values
were not tabulated on days 14 to 21 of lactation.
418-014:PAGE B-16
001341
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 (%)
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
MEAN+S.D .
22.4 + 0.5
22.0 + 0.0
NUMBER OF DAMS DELIVERING
DAY 22
N (%)
DAY 23
N (%)
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
DAY(S) = DAY(S) OF GESTATION a. Excludes values for dams which had litters that were not cross-fostered. b. Calculated as the time (to the nearest tenth of a day) elapsed between confirmed mating (arbitrarily defined as 0 hour) and
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.
418-014: PAGE B-17
001342
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 OP DELIVERY PER PUP PER LITTER (MINUTES)
TOTAL DELIVERY PERIOD OBSERVED
(MINUTES) b
MEAN+S.D.
TOTAL OR PARTIAL DELIVERY
PERIOD OBSERVED
(MINUTES) b
MEAN+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 MEAN+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)
K 4.3)
PUPS FOUND DEAD (PRECROSS-FOSTERING)
LIVE LITTER SIZE C
N / N (%) MEAN+S.D .
1/373( 0.3) 16.2 + 1.8
1/344 ( 0.3) 14.9 + 1.8
FOSTER LITTER SIZE
MEAN+S.D.
15.2 + 1.9
15.6 + 1.9
( ] = 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 cross-fostering.
418-014:PAGE B-18
001343
418-014:PAGE B-19
001344
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE IS (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
MEAN+S.D .
16.4 + 1.6
15.9 + 2.1
14.8 + 1.9
FOSTER LITTER SIZE
MEAN+S.D.
15.1 + 1.7
15.9 + 2.1
14.8 + 1.9
DAMS WITH ALL PUP DYING DAYS 1-4 POSTPARTUM (AFTER CROSS-FOSTERING)
N (%)
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-20
001345
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 MEAN+S.D .
166 15.1 + 1.7
191 15.9 + 2.1
177 14.8 + 1.9
PUPS FOUND DEAD OR PRESUMED CANNIBALIZED
DAY 1 DAYS 2- 4 DAYS 5- 7 DAYS 8-14 DAYS 15-21
N / N (%) N / N (%) N / N (%) N / N (%) N/N (1)
0/166( 15/166(
1/109( 0/1081 0/108(
0.0) 9.0) 0.9) 0.0) 0.0)
0/191( 3/191( 0/120( 0/120{ 0/120(
0.0) 1.6) 0.0) 0.0) 0.0)
0/177( 0.0) 34/1771 19.2)
0/107( 0.0) 0/107( 0.0) 0/107( 0.0)
VIABILITY INDEX f
% N/N
91.0 151/166
98.4 188/191
B0.8 143/177
LACTATION INDEX g
t N/N
99.1 108/109
100.0 120/120
100.0 107/107
DAY(S) = DAY(S) POSTPARTUM a. Vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. Vehicle control group dams cross-fostered with vehicle control litters. c. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. e. Excludes values for dams which had litters that were not cross-fostered. f. Number of live pups on day 4 (preculling) postpartum/number of pups cross-fostered on day 1 postpartum. g. Number of live pups 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
0/18M 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
418-014:PAGE B-21
001346
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
MEAN+S.D .
1S.1 + 1.7
15.9 + 2.1
14.8 + 1.9
DAY 4 PRECULLING
MEAN+S.D.
13.7 + 2.3
15.7 + 1.8
11.9 + 4.6
DAY 4 POSTCULLING MEAN+S.D .
9.9 + 0.3
10.0 + 0.0
8.9 + 2.9
DAY 7
MEAN+S.D .
9.8 + 0.6
10.0 + 0.0
8.9 + 2.9
DAY 14
MEAN+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 croBS-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
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 16 (PAGE 3): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - 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 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
[ HJf 46.0 + 16.9
l H]f 46.0 + 16.9
l Ulf 46.0 + 16.9
( Ulf 46.0 + 16.9
[ Ulf
DAY 1
MEAN+S.D .
15.1 + 1.7
15.9 + 2.1
14.8 + 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
[ H]f 9.7 + 0.6
[ lllf 9.7 + 0.6
[ ll)f 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 had 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 f 0.0 50.0 0.0 50.0 + 0.0
16.4 + 1.6 16.3 1.6 10.0 0.0 1 10.0 + 0.0 10.0 f 0.0 10.0 + 0.0
418-014:PAGE B-22
001347
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)
o UJ
DAY ig
MEAN+S.D .
5.6 +
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 . MEAN+S.D . MEAN+S.D . MEAN+S.D .
7.1 + 0.5 7.3 + 0.5 12.3 + 0.9 26.7 + 1.7 41.9 + 1.0
8.3 + 0.8 8.5 + 0.8 14.1 + 0.9 29.0 + 1.5 46.3 + 2.5
7 . 2 + 0.8
[ H]f 7.3 + 0.8
[ lljf 11.6 + 1.3
( Ulf 24.6 + 2.4
[ 111 f 38.9 + 3.8
( Ulf
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-fostered. f. Excludes values for dam 18974, which had no surviving pups on day 4 postpartum. g. Day 1 pup weights for Subsets A and C reflect pups of test article treated dams. Day 1 pupweights forSubsets B and D
reflect pups of vehicle treated dams. The effects of cross-fostering are not reflected inthese pup body weights.
418-014: PAGE B-23
001348
418-014:PAGE B-24
001349
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)
TRANSIENT CLINICAL OBSERVATIONS: e
A Ia 0 (VEHICLE) 13
B Ib 0 (VEHICLE) 12
C 11 c 1.6 12
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 group litters. b. Vehicle control group dams cross -fostered with vehicle control litters. c. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. e. Tabulation restricted to adverse observations; all other pups appeared normal.
D II d 1.6 13
18/ 1 1/ 1 1/ 1 0/ 0
0/ 0 0/ 0
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
418-014:PAGE B-25
O
o o
oo oo oo
001350
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)
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(t)
PUP INCIDENCE
N (%)
If 11.1) 1( 1.3)
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 croBs-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.
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 1890S 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: ABRASION (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.
418-014PAGE B-26
001351
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 19 (PAGE 2): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS
DOSAGE GROUP I
0 (VEHICLE) MG/KG/DAY
RAT
DESCRIPTION
18928 18929 18930 18931 18932 18933 18934 18935 1B936
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 CHROMORHINORRHEA 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.
418-014PAGE B-27
001352
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 tt
DESCRIPTION
1B943
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 )
DL( 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
001353
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 RAT H
1.6 MG/KG/DAY 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)
D M 1- 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
0013S4
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
I. 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
1: DL 22
P
86
ALL TISSUES APPEARED NORMAL.
18908
A 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 ? d l 14
P
80
ALL TISSUES APPEARED NORMAL.
18912 :> J. DL 22
P
87
ALL TISSUES APPEARED NORMAL.
18913
7 DL 14
P
80
ALL TISSUES APPEARED NORMAL.
18914
: DG 25
NP
68
ALL TISSUES APPEARED NORMAL.
18915
1 DL 14
P
76
ALL TISSUES APPEARED NORMAL.
18916
i DG 25
P
70
ALL TISSUES APPEARED NORMAL.
i c UTERINE CONTENTS: ONE IMPLANTATIONS (ONE DEAD FETUS) .b
18917
.A DL 22
P
87
ALL TISSUES APPEARED NORMAL.
18918
U, DL 22
P
87
ALL TISSUES APPEARED NORMAL.
18919 U M DL 22
P
87
ALL TISSUES APPEARED NORMAL.
18920 1 DL 22
P
86
ALL TISSUES APPEARED NORMAL.
18922
K.DL 22
P
85
ALL TISSUES APPEARED NORMAL.
18923 X N:DL 22
P
87
ALL TISSUES APPEARED NORMAL.
18924
: DL 22
P
87
ALL TISSUES APPEARED NORMAL.
18925 u u DG 26
NP
69
ALL TISSUES APPEARED NORMAL.
18926
DL 14
P
81
LEFT INGUINAL AREA: MASS (2.5 CM X 2.0 CM X 1.5 CMl.C
' ALL OTHER TISSUES APPEARED NORMAL.
go
w
in
in
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. b. The fetus had a large, black right: bindlimb and a black tail. Evidence of compression on the abdomen and hindlimbs. c. First observed at necropsy.
418-014:PAGE B-30
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 a p p e a r e d 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-31
001356
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 FETUSES.b
18978
DG 25
NP
69
ALL TISSUES APPEARED NORMAL.
P = PREGNANT NP = NOT PREGNANT DS = DAY OP 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. b. Appeared normal for their developmental ages and moderate degree of autolysis at time maternal death occurred.
418-014:PAGE B-32
001357
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 - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS
RAT #
DOSAGE GROUP I
DAY 1 2 3 4
18901 18902 18903 18904 1890S 18906 18907 18908 18909 18910 18911 18912 18913 18914 1891S 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
5678
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.
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 .
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.
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.
15
260. 256. 269. 245. 237. 284. 259. 242. 247. 248. 257. 257. 254. 280. 262. 265. 262. 262. 250. 267. 250. 272. 253. 273. 247. 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
001358
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 GENERATION FEMALE RATS
RAT ft
DOSAGE GROUP I
0 (VEHICLE) MG/KG/DAY
DAY 1 2 3 4 5 6 7 8 9 10
18927 18928 18929 18930 18931 18932 18933 18934 1893S 18936 18937 18938 18939 18940 18941 18942
231. 24S. 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.
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.
237. 263. 231. 258. 257. 243 . 227. 253. 244 . 254. 238. 261. 244 . 265. 247. 254 .
229. 249. 233. 262. 251. 249. 231. 260. 239. 252. 239. 253. 250. 265. 249. 247.
DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G) .
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 15 16
232. 254. 236. 270. 256. 258. 233 . 273 . 255. 261. 248. 269. 258. 278. 255. 254 .
242. 260. 244 . 264. 262. 267. 233. 272. 261. 255. 240. 272. 252. 269. 254. 261.
243. 265. 242. 268. 265. 265. 242. 268. 261. 261. 242. 278. 260. 278. 251. 265.
418-014:PAGE B-34
001359
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PAGE 3): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS
RAT tt
DOSAGE GROUP I
0 (VEHICLE) MG/KG/DAY
DAY
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
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. 250. 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.
21 22 23 24 25
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.
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.
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.
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.
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.
DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G).
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
001360
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 #
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
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.
25
243 . 277. 260. 283 . 270. 265. 258. 294 . 271. 276. 254. 283. 279. 301. 270. 271.
26
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 29 30
246. 284 . 264. 286. 280. 278. 256. 292. 275. 284. 249. 290. 278. 295. 275. 274.
244 . 278. 260. 291. 278. 274. 262. 297. 274. 282. 252. 290. 280. 300. 275. 272.
243. 277. 257. 288. 278. 279. 260. 304. 272. 285. 256. 279. 278. 302. 274. 266.
31 32
246. 285. 263. 285. 283. 280. 259. 300. 275. 278. 250. 292. 276. 294. 273 . 276.
250. 288. 266. 289. 285. 282. 263. 298. 275. 286. 259. 293. 282. 302. 278. 279.
418-014:PAGE B-36
0013S1
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 21 (PAGE S) : BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS
RAT
DOSAGE GROUP I
0 (VEHICLE) MG/KG/DAY
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
33
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.
34
284 . 283. 292. 269. 252. 317. 287. 261. 275. 264. 277. 286. 270. 307. 299. 277. 282 . 273. 274 . 285. 262. 306. 267. 298. 273. 293 .
35
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.
36
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.
37
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.
38
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.
39
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.
40
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.
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.
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.
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.
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 ft
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. 285. 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 on 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
001383
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
i234
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 14 15
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.
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 .
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. 250. 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
001364
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 21 (PAGE 8): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS
RAT #
DOSAGE GROUP II
1.6 MG/KG/DAY
DAY 1 2 3 4 5 6 7 8 9 10 11 12 13
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.
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 .
246. 245. 243. 250. 247. 263 . 256. 229. 235. 251.
247. 243 . 245. 254. 244. 259. 257. 242. 238. 255.
244 . 245. 239. 259. 242. 263 . 250. 241. 242. 253.
252. 254. 245. 260. 249. 271. 263. 241. 243 . 254.
259. 254. 252. 264. 252. 272. 266. 243 . 240. 252.
256. 249. 250. 262. 249. 269. 266. 249. 245. 261.
DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G).
14 15 16
253 . 256. 246. 267. 249. 276. 263 . 256. 243 . 260.
262. 261. 251. 273. 256. 278 . 274 . 258. 247. 259.
261. 258. 253. 272. 259. 276. 280. 258. 247. 256.
418-014:PAGE B-40
001385
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 H
DOSAGE GROUP II
1.6 MG/KG/DAY
DAY 17 18 19 20
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
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).
21
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.
22
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.
23
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. 257. 268 . 246. 283 . 267. 261. 255. 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. 268. 279. 262. 275. 251. 289. 278. 261. 263. 272. 265. 278. 255. 303. 258. 287. 257. 261. 248. 275. 276. 278.
29 30 31 32
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.
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.
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.
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
001366
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 ft
DOSAGE GROUP ii
1.6 MG/KG/DAY
DAY 17 18 19 20 21 22 23 24 25
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.
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 .
274 . 266. 261. 282 . 262. 282. 291. 264 . 252. 266.
271. 263. 264. 281. 265. 279. 290. 265. 261. 273.
DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G) .
26 27
271. 272. 263. 278. 262. 281. 287. 271. 263 . 273.
274. 274. 268. 285. 266. 289. 290. 270. 262. 270.
28 29 30 31 32
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 .
279. 279. 274. 295. 269. 285. 299. 275. 262. 274.
418-014:PAGE B-42
001367
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 It
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
252 . 295. 255. 278. 274. 284 . 263. 266. 256. 295. 280. 270. 263. 281. 275. 260. 261. 303. 270. 288. 263 . 271. 246. 280. 284 . 299.
35
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 37
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.
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 39
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.
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. 24S. 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
89CT00
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 tt
DOSAGE GROUP ii
1.6 MG/KG/DAY
DAY 33 34 35 36 37 38 39 40 41 42 43a
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.
291 . 281. 276. 298. 275. 284. 302. 278. 267. 265.
287. 274 . 274 . 295. 270. 278. 305. 274. 272. 276.
285. 281. 275. 289. 269. 286. 301. 280. 276 . 276.
284 . 282. 275. 291. 275. 288. 301. 280. 273. 275 .
DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation.
418-014:PAGE B-44
001369
418-014:PAGE B-45
001370
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 ft
DOSAGE GROUP I
0 (VEHICLE) MG/KG/DAY
PREGNANCY STATUS DAY 0 1 2
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.
260 .
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 .
3
312 303 316 292
366 319 287 304 286 310 319 306 329 335 296 308 299 304 307 265 325 291 346 288
4
321. 308. 325. 294 .
370. 318, 292 , 308. 293 . 318. 321, 300 . 335 , 335. 310. 314 . 301. 306. 310. 267 . 329, 296 . 355. 284 .
5
316. 314. 329. 299.
373. 330. 291. 310. 295. 316. 328 . 300. 342. 340. 306. 317. 312 . 315. 315. 263. 336. 298. 347. 275.
6
323. 316. 333. 301.
378. 329. 304 . 317. 299. 322. 328. 302. 350. 346. 308. 323 . 319. 320. 316. 259. 341. 299. 351. 289.
7
327. 318. 330. 307 .
377. 327. 302. 320. 301. 327. 337. 314. 350 . 345. 310. 326. 322. 320. 319. 262. 345. 302 . 360. 285.
8
326. 317. 335. 304.
384 . 333 . 307. 321. 304 . 330. 335. 309. 350. 354. 317. 328. 321. 329. 317. 263. 352. 301. 356. 287.
9
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 ALL WEIGHTS WERE RECORDED IN GRAMS (G).
a. Dam 18916 did not deliver a litter; only one dead fetus was present in utero on day 25 of gestation
10
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. 31 g . 34 5 361 334 343 382 . 329 348 340 353 . 344 . 266 . 378. 321. 376. 291.
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) 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 3 4 5
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.
2B8 .
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.
328 . 298.
308. 332. 314 . 307. 300. 340. 306. 278. 289. 329. 323. 346 . 326. 312 .
332. 303.
320. 334 . 320. 312. 306. 341. 319. 277. 294 . 328 . 324 . 357. 327. 315.
332. 304.
317. 334. 322. 315. 303. 351. 321. 278. 300. 334 . 326. 369. 333 . 321.
> PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION
ALL WEIGHTS WERE RECORDED IN GRAMS (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.
35fi 352. 356. 344 . 335 373 . 340 174 317 366 356 390 364 346. ----
418-014:PAGE B-46
418-014:PAGE B-47
001372
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 22 (PAGE 3): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS
RAT tt
DOSAGE GROUP I
0 (VEHICLE) MG/KG/DAY
PREGNANCY STATUS DAY 13 14 15
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 .
325.
333.
341.
393.
388.
394.
280.
288.
294.
16
374 309 392 356
440 380 363 378 333 374 388 368 342 426 334 372 369 382 372 267 404 348 398 289
17
379. 309. 402. 371.
460. 396. 377. 394 . 342. 392. 406. 383 . 346 . 436. 334 . 386. 378. 394 . 383. 263. 415. 359. 403 . 287.
18
400. 307. 426. 387.
483. 414 . 397. 395. 346. 409. 427. 404. 349. 452. 340. 396. 395. 413 . 406. 258. 431. 376. 421. 286.
19
417. 300. 452. 406 .
499. 436. 413 . 414 . 355. 424 . 444 . 409 . 346. 478. 347 . 412. 413. 434. 428. 263. 442. 394 . 442 . 287.
20
436. 304. 470. 427.
523. 456. 443 . 427. 357. 455. 467. 434. 340. 491. 341. 428. 425. 4S4 . 434. 270. 465. 405. 468. 298.
21
455. 313. 488. 450.
526. 478. 459. 445. 369. 464 . 480. 443 . 350. 525. 341. 444 . 442. 463. 451. 271. 477. 412. 476. 298.
22
314. 304 .
429. 329. 457. 347. 336 . 438. 431.
268.
294.
P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G).
a. Dam 18916 did not deliver a litter; only one dead fetus was present in utero on day 25 of gestation
23 306.
351. 322. 277. 292.
24 ~-
303.
25 --
315.
350. 324 .
350
327
280.
286
288.
290.
PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 22 (PAGE 4) : MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS
RAT #
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.
390.
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.
= PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION
ALL WEIGHTS WERE RECORDED IN GRAMS (G).
19
443 . 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-48
001373
i
ffrLtOQ
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 22 (PAGE 5): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS
RAT n
DOSAGE GROUP II
PREGNANCY STATUS DAY 0 1 2
18943 P 18944 NP 18945 NP 18946 P 18947 P 18948 P 18949 NP 1895Q 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. 292.
286. 300.
293 . 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.
28.2.
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. 2B4 . 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.
418-014:PAGE B-49
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 1 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
418-014.PAGE B-51
00137$
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 20
394 . 321.
417. 325.
418. 405. 443 . 277. 410. 375. 420. 413. 263 . 296. 380. 401. 421. 376. 438. 401. 288. 378. 374. 407.
444 . 412 . 458. 280. 419. 393 . 430. 444 . 266 . 269. 395. 413 . 426 . 394 . 449. 417. 290. 391. 391 . 411.
425.
445.
424 . 327.
450. 431. 470. 271. 442. 406. 446 . 442. 254 . 273. 412. 420 . 443. 406. 461. 436. 288. 406. 399.
457.
325.
272 .
266 . 274. 416.
293 .
322. 276. 255. 269.
294 .
329. 275. 254 . 271.
297.
328. 278. 264 . 277.
293.
& TOO
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 IS 16 11 18
18968 P 18969 P 18910 P 18911 P 18912 P 18913 P 18914 P 18915 NP 18916 P 18911 P 18918 NP
342. 345. 321. 322 . 365. 321 . 354. 326 . 338. 341 . 218.
351. 350. 333. 320. 365. 321. 361. 315 . 343 . 345 . 281.
366. 364. 334. 324. 365. 329. 314 . 316. 352 . 358. 291.
313. 312. 346. 319. 383 . 341. 378. 324 . 360 . 310. 284 .
381. 386. 362. 332. 398. 355. 396. 311. 312. 380. 214 .
401. 415. 314. 340. 422. 381. 410. 313. 389. 394 . 281.
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. 431. 398. 433 . 318 . 404 . 414 . 291 .
20
441. 459. 406. 354. 460. 422. 440. 329. 423. 430. 288 .
21
452. 412. 406 . 355. 468. 420. 469. 309. 456. 461. 283.
22
356. 309. 295.
23
318. 291. 294.
24
320. 291.
25
311. 290.
418-014:PAGE B-52
&LCTOO
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 H
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.
a
345. 341. 336. 362. 360. 342. 368. 354. 369. 335. 358. 357. 354.
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 waB 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
37Q
314 . 371 374 . 361.
418-014:PAGE B-53
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 23 (PAGE 2) MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS
RATS ASSIGNED TO CROSS-FOSTERING
RAT ))
DOSAGE GROUP I a
0 (VEHICLE) MG/KG/DAY
1B901 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 . Jbfa . 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. 448. 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. 366. 328. 363. 347. 343 . 392.
"*-- '
i
418-014:PAGE B-54
CTOO
6& C T00
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 23 (PAGE 3) : MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS
RATS ASSIGNED TO CROSS-FOSTERING
RAT #
DOSAGE GROUP II a
1.6 MG/KG/DAY
SUBSET C
DAY 1 2 3 4 5 6 7 a 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 group dams cross-fostered with 1.6 mg/kg/day dosage group 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. 346366. 341. 330. b 321. 368359. 366. 340. 314.
418-014:PAGE B-55
418-014:PAGE B-56
00138#
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
18946b 18950 18951 18952 18953 18958 18959 18961 18963 18965 18972 18976 18977b
18946b 18950 18951 18952 18953 18958 18959 18961 18963 18965 18972 18976 18977b
DOSAGE GROUP II a
1.6 MG/KG/DAY
SUBSET D
DAY 1 2 3 4 5 6 7 8 9
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
3S0.
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 IG) . a. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. b. Litter was not cross-fostered; values excluded from group averages.
10
346. 335. 311. 360. 339. 331. 309. 330. 328. 332. 368. 349.
11
352. 337. 309. 360. 348. 329. 318. 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
'tfsCTOO
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-57
fsC TO O
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING 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.
418-014:PAGE B-58
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 24 (PAGE 1): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS
RAT
DOSAGE GROUP I
0 (VEHICLE) MG/KG/DAY
DAYS
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
1- 8
147. 123.
c c 133. 144. 147. 116. 132. 122. 137. 136. 116. 152. 147. 145. 142. 127. 123 . 136. 130. 129. 139. 159. 142. 143.
8- 15 15- 22 22- 29 29- 36 36- 43a
146. 135. 152. 131. 138. 149. 148. 129. 138.
c 133. 139. 139. 153. 148. 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.
418-014PAGE B-59
ooias
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 It
DOSAGE GROUP I
0 (VEHICLE) MG/KG/DAY
DAYS 1- 8 8- 15 15- 22 22- 29 29- .36 36 43a
18927 18928 18929 18930 18931 18932 18933 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 study. c. Spilled feed precluded the calculation of this value.
418-014:PAGE B-60
h* CO
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295 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- 15 15- 22 22- 29 29- 36 36- 43a
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
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. 111. 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 HEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. Spilled feed precluded the calculation of this value. c. Value appeared incorrectly recorded and was excluded from group averages.
418-014PAGE B-61
$SCTOO
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 24 (PAGE 4): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS
RAT #
DOSAGE GROUP 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
00138JI
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 #
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. 2S.
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.
: 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 - 1
24. 22. 27. 25. 26. 19. 24. 24. 29. 27. 34 . 30. 23. 20. 24. 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
&SCTOO
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 -
.- 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. 23. 22. 21.
26. 26. 26. 26. 24. 24.
MATING NOT CONFIRMED
24 . 26. 24. 27. 23 . 26.
27. 28. 29. 27. 28. 24.
24 . 27. 26. 29. 28. 30.
19. 24. 24 . 25. 22. 23.
26. 25. 25. 25. 24 . 24.
27 . 27 . 29. 26. 30. 30.
14 . 18. 22. 28. 27 . 26.
16 . 16.
16 . 15.
13 . 14 .
20 .
26.
24 . 23 . 23.
24 .
23. 28. 25. 25. 27. 25 .
25. 26. 22. 26. 26 . 24 .
23. 26. 31. 30. 34 . 23 .
23 . 24 . 27. 27. 24 . 24 .
24 . 23 . 22 . 24 . 27 . 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 .
1- 9
22. 23.
29. 30. 28. 28. 29. 28. 25. 22. 20. 25. 26. 29. 27. 25.
9-10
27. 22.
26. 32. 30. 26. 24 . 32 . 24 . 21. 23. 28. 25. 33. 26. 24 .
10 - 11 11 - 12 12 - 13
25. 26. 26. 26. 23. 26.
32. 29. 31.
27. 31. 25.
31. 23. 32.
25. 29. 24.
24. 26. 24.
26 . 24 . 31.
25. 24. 30.
28. 21. 20.
24 . 22.
24.
29. 26. 30.
28. 24. 21.
27. 30. 27.
28. 23 . 28.
27. 26. 26.
418-014:PAGE B-64
001.38
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 1 3 - 1 4 14 - 15 15 - 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 18902 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. 18.
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
0013 < 1 0
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 DATA - Fo GENERATION FEMALE
RAT #
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. 27. 23. 40. 32. 16.
22. 13. 22. 20. 27. 17. 34. 25.
MATING NOT CONFIRMED
31. 13. 27. 28. 27. 22. 36. 27.
10. 19. 20. 19. 12. 23. 19.
3. 11. 23. 18. 16.
27. 14 . 29. 27. 31. 20. 32 .
25. 24 . 19. 23. 25. 26. 19. 28.
25 . 23 . 32 . 25. 25. 32. 28.
27. 31. 27. 19. 26. 28. 27. 18.
27 . 23 . 30 . 28. 28. 21. 29.
26. 29. 25. 24 . 22. 27. 19. 19.
20. 12 . 21. 22. 24 . 19. 28. 25.
28.
13.
26 .
26.
26.
20 . 29.
28. 24 . 25. 31 . 18. 23 . 25. 28.
17. 20. 22. 23. 17. 28 . 26.
7.
23. 19. 31. 26. 29. 22. 36. 29.
27. 15. 17. 31. 26. 19. 28. 13 .
P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G).
418-014:PAGE B-66
00139/
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 25 (PAGE 5): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - 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 1 0 - 1 1 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.
418-014:PAGE B-67
'tcTOO
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 DATA - Fo GENERATION FEMALE RATS
RAT ((
DOSAGE GROUP II
1.6 MG/KG/DAY
PREGNANCY STATUS DAYS 0 - 1 1 - 2 2 - 3 3 - 4 4 - 5
5- 6 6 - 7 7-8
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.
25. 25. 22. 23. 23. 17. 25. 23 . 27. 26. 21.
24. 28. 23. 20. 25. 19. 25. 22. 22 . 26. 24 .
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.
8- 9
24. 23. 25. 20. 24. 19. 25. 24 . 31. 22. 20.
9 - 10 10 - 11 11 - 12 12 - 13
21. 25. 25. 27. 27. 29. 26. 31. 21. 24. 23. 23. 23. 21. 22. 20. 25. 26 . 27. 30. 23. 23. 21. 25. 23. 24 . 29. 27. 26. 26. 23. 19. 20. 26. 27. 28. 25. 24 . 27. 28. 19. 22. 10. 12.
418-014:PAGE B-68
6 CT0 0
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 25 (PAGE 7): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS
RAT tf
DOSAGE GROUP II
1.6 MG/KG/DAY
PREGNANCY STATUS DAYS 13 - 14 14 - 15 15 - 16 16 - 17 17 - IB 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. 5. 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. 28. 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. 12.
7. 18.
15. 14 . 18. 15. 18.
18 . 19.
21. 16 .
5. 7.
8.
16. 12. 14. 15.
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
^6C T 00
PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 25 (PAGE 8): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS
RAT #
DOSAGE GROUP 11
1.6 MG/KG/DAY
PREGNANCY STATUS DAYS 13 - 14 14 - 15 15 - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 21 - 22 22 - 23 23 - 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. ii. 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
6 C T0 0
PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 26 (PAGE 1): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS
RATS ASSIGNED TO CROSS-FOSTERING
DAYS 1 - 4 4 - 7 7 - 10 10 - 14a
RAT #
18903 18907 18908 18909 18918c 18920 18924 18929 18934 18936c 18937 18941 18942
DOSAGE GROUP I b
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.
0 (VEHICLE) MG/KG/DAY
232. 235. 243 . 207. 264. 226. 218. 257. 215. 183 . 250 . 219. 239.
SUBSET A
RAT #
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
6C T00
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 ft
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. Litters were not cross-fostered; values excluded from group averages.
i
418-014:PAGE B-72
00139 7
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 26 (PAGE 3): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS
RATS ASSIGNED TO PHARMACOKINETIC EVALUATION
RAT H
DAYS 1 - 4 4 - 7 7 - 1 0 DOSAGE GROUP I
10 - 14 0 (VEHICLE) MG/KG/DAY
18910 18911 18913 18915 18926 18927 18939 18940
RAT It
61. 107.
68. 88. 97. 66. 86. 82.
73. 149. 134 . 138. 131. 119. 120. 106.
114 . 148. 140. 216. 181. 137. 154 . 141.
DOSAGE GROUP II
145. 251. 261. 246. 300. 231. 230. 265 .
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) .
418-014:PAGE B-73
00139g
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 0
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
S5
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 .
16946
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 15 19 20 16 12 15 16 17 14
16 , 15
6 10 78 8 11 11 9 97 75 5 10 6 10 7 10 68 79 13 2
6 10 78 8 10 11 8 97 75 5 10 69 7 10 68 79 13 2
55 55 55 55 55 55 55 55 55 55 55 82
55 55 55 55 55 55 55 55 55 55 55 82
55 55 55 55 55 55 55 55 55 55 55 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-74
6C T00
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER) T-6295.13)
TABLE 27 (PAGE 2): NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY 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
TOTAL IMPLANTATINS
N'
FOSTER LITTER #
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 II c
1.6 MG/KG/DAY
SUBSET C
18943 18947 16948 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 IS 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
IS
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 0 13 0 14 0 16 0 13 0 18 0 17 0 15 0 12
17 18918 17 18929 13 18941 17 18920 18 18908 14 18937 15 18934 16 18909 13 18907 18 18942 18 18924 15 18903 16 18936
15
98
3i
3i
21
21
18
8 10
8 10
55
55
55
14
86
B6
55
55
5S
17
89
89
S5
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 8
55
55
55
17
6 11
6 11
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
--
418-014:PAGE B-75
OQ/r too
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
I 4 7 14
MF
MF
MF
MF
MATERNAL DOSAGE GROUP I
0 (VEHICLE) MG/KG/DAY
18910 18911 18913 18915 18926 18927 18939 18940
505 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 a8 78 10 6
32 11 4 14 2
4 il 66 88 78 10 6
32 11 4 14 2
4 il 66 87 78 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
|0 tT 0 0
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 DATA Fo GENERATION FEMALE RATS
RATS ASSIGNED TO CROSS -FOSTERING
RAT/ LITTER NUMBER
DURATION OP 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 11.7 f 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 group dams cross-fostered with 1.6 mg/kg/day dosage group litters. d. Vehicle control group dams cross-fostered with vehicle control litters. e. Litters were not cross-fostered; values excluded from group averages. f. Partuition period only partially observed.
418-014:PAGE B-77
O'o'TOO
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 DATA 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 II c
1.6 MG/KG/DAY
SUBSET C
18943 18947 18948 18957 1B960 18964 18967 18968 18969 18970 18973 18974
21.5 21.5 21.6 21.6 21.6 21.8 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 21.7 21.6
22 12.3f 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
i
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 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. f. Partuition period only partially observed.
418-014:PAGE B-78
fo frT O O
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13)
TABLE 28 (PAGE 3): DURATION OF GESTATION AND AVERAGE DURATION OF DELIVERY (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 aa 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.
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 29 (PAGE 1) : PUP BODY WEIGHT LITTER AVERAGES FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION LITTERS
RATS ASSIGNED TO CROSS-FOSTERING
RAT/ FOSTER LITTER tt
DAY 1 M FT
DAY 4a M FT
DAY 4b M FT
DAY 7 MF
T
DAY 14 M FT
DAY 21 MFT
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
5.6 5.2 5.4 5.9 5.9 5.9 6.1 5.9 6.0 10.5 9.6 10.0 24.7 23.2 24.0 41.1 38.2 39.7
6.1 5.8 5.9 7.8 7.4 7.6 7.0 7.6 7.7 13.0 12.6 12.8 27.9 27.1 27.5 44.2 42.6 43.4
5.8 5.4 5.5 7.1 7.0 7.0 7.1 7.2 7.2 12.6 11.7 11.9 26.8 26.2 26.4 42.1 39.9 40.6
5.5 5.5 5.5 7.4 7.2 7.3 7.7 7.7 7.7 13.4 13.4 13.4 28.1 27.8 28.0 42.7 40.5 41.6
5.6 5.1 5.3 10.0 9.2 9.6 10.0 9.1 9.6 16.5 15.0 15.8 33.0 30.6 31.8 50.0 49.9 50.0
5.3 5.2 5.3 6.7 7.0 6.8 7.3 7.0 7.1 12.3 11.8 12.0 25.0 24.5 24.7 41.4 40.5 40.9
5.7 5.5 5.6 7.5 6.7 7.1 7.5 7.3 7.4 12.5 12.0 12.3 25.9 25.3 25.6 41.2 40.9 41.0
5.6 5.3 5.4 7.6 7.2 7.4 8.1 7.4 7.8 13.7 12.5 13.1 30.2 27.9 29.0 47.2 43.8 45.5
5.8 5.4 5.6 7.4 7.0 7.2 7.6 7.0 7.3 13.0 12.4 12.7 27.6 26.3 27.0 42.0 40.2 41.1
6.2 5.8 6.1 8.7 7.7 8.3 8.6 7.7 8.2 13.1 12.0 12.7 25.1 23.0 24.2 39.9 36.6 38.6
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 40.3 41.0 40.7
6.2 5.9 6.1
8.2 6.9 7.6
8.2 6.9 7.6 12.8 12.7 12.8 28.2 29.2 28.6 44.5 45.4 44.8
5.3 5.1 5.2 7.0 7.0 7.0 7.2 7.3 7.3 12.4 12.5 12.4 28.0 28.4 28.2 41.7 41.6 41.6
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 .a 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 50.0 46.4 48.2 8.0 7.4 7.7 14.2 12.6 13.4 28.9 26.5 27.7 44.4 41.5 43.0 8.4 8 .1 8.3 13.5 13.4 13.4 28.7 28.3 2B.5 49.9 47.9 48.9 7.0 6.3 6.6 12.5 12.7 12.6 29.8 27.5 28.6 48.6 44.7 46.7 8.6 8.4 8.5 13.8 13.6 13.7 28.9 28.4 28.6 45.3 45.7 45.5 9.3 9.3 9.3 15.6 15.1 15.4 30.9 29.9 30.4 4 9 0 47.8 48.4 8.9 8.9 8.9 15. S 15.1 15.3 30.8 31.0 30.9 48.7 48.4 48.5 9.8 8.8 9.3 16.1 14.3 15.2 30.5 28.1 29.3 50.4 46.1 48.3 8.2 8.2 8.2 13.8 13.8 13.8 30.6 30.5 30.6 48.9 47.7 48.3 9.2 8.7 8.9 14.5 13.9 14.2 27.4 26.9 27.2 43.7 43.5 43.6 8.5 8.1 8.3 13.5 13.0 13.2 26.2 25.9 26.0 42.3 41.5 41.9 8.7 8.4 8.6 13.8 13.8 13.8 29.2 29.0 29.1 44.5 43.8 44.3
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. Poatculling
c. Vehicle control group dams cross-fostered with 1.6 mg/kg/day dosage group litters.
d. Litters were not cross-fostered; values excluded from group averages.
e. Vehicle control group dams cross-fostered with vehicle control litters.
418-014:PAGE B-80
(b frTO O
PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 29 (PAGE 2) : PUP BODY WEIGHT LITTER AVERAGES FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION LITTERS
RATS ASSIGNED TO CROSS-FOSTERING
RAT/
FOSTER LITTER It
DAY 1 M FT
DAY 4a M FT
DAY 4b M FT
DAY 7 M FT
DAY 14 M FT
DAY 21 MFT
MATERNAL DOSAGE GROUP II c
iL.6 MG/KG/DAY
SUBSET C
18943 18947 18948 18957 18960 18964 18967 18968 18969 18970 18973 18974
5.8 5.6 5.7 8.0 8.1 8.0 8.0 8.2 8.1 12.9 13.0 13.0 28.4 28.1 28.2 43.1 42.0 42.5
5.8 5.6 5.7 7.6 7.1 7.4 7.8 7.2 7.5 12.5 11.7 12.1 26.4 24.2 25.3 43.1 39.7 41.4
5.8 5.4 5.7 6.3 5.8 6.2 6.3 5.8 6.2 10.1 9.0 9.8 23.7 20.8 23.0 36.8 33.6 36.0
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 39.7 39.8 39.8
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 35.9 32.4 33.4
6.1 6.0 6.0 6.6 6.7 6.6 6.6 6.9 6.8 10.3 11.0 10.7 21.4 22.3 21.8 33.9 35.0 34.4
5.8 5.5 5.7 8.0 7.2 7.6 8.3 7.0 7.7 14.0 11.3 12.6 29.5 24.9 27.2 47.1 41.1 44.1
6.6 6.2 6.3 10.4 8.7 8.9 10.4 8.7 8.9 15.4 13.4 13.6 30.4 26.6 27.1 50.7 42.9 43.8
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 40.5 38.2 39.4
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 39.9 33.0 36.4
5.6 5.4 5.6 7.4 7.2 7.3 7.5 7.2 7.4 11.4 10.9 11.2 23.0 22.9 23.0 36.6 36.6 36.6
5.7 5.3 5.4 NO SURVIVING PUPS
MATERNAL DOSAGE GROUP II d
1.6 MG/KG/DAY
SUBSET D
18946e 18950 18951 18952 18953 18958 18959 18961 18963 18965 18972 18976 18977e
6.9 6.4 6.7 7.1 6.3 6.7 7.8 7.5 7.6 6.4 5.9 6.1 7.0 6.5 6.8 7.2 6.4 6.7 6.0 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 48.8 36.3 44.6 8.1 7.3 7.7 8.4 7.6 8.0 13.3 12.3 12.8 26.9 25.4 26.1 44.5 39.7 42.1 8.8 8.4 8.6 9.0 8.4 8.7 12.6 11.2 11.9 24.2 21.6 22.9 39.4 36.5 38.0 8.2 7.4 7.8 8.7 7.6 8.2 14.8 13.3 14.0 30.7 28.1 29.4 47.1 43.4 45.3 8.2 8.2 8.2 8.6 8.5 8.6 12.7 11.9 12.3 25.4 24.8 25.1 41.6 40.3 41.0 9.2 8.1 B .6 9.1 8.4 8.7 12.9 12.3 12.6 24.7 24.0 24.4 40.2 39.5 39.8 8.2 7.6 8.0 8.0 7.7 7.8 13.0 12.5 12.7 27.1 26.2 26.6 42.8 42.1 42.5 8.3 7.7 7.9 8.3 8.5 8.4 13.3 13.1 13.2 26.3 25.7 26.0 42.0 40.1 41.1 7.1 6.4 6.8 7.3 6.6 6.9 12.2 11.7 12.0 26.8 25.9 26.4 43.2 41.7 42.5 8.1 7.6 7.8 8.2 7.7 8.0 12.7 12.3 12.5 2 5 6 24.9 25.2 44.0 42.3 43.1 9.0 8.1 8.5 9.0 8.3 8.7 14.8 14.0 14.4 31.4 29.9 30.6 46.1 43.7 44.9 7.4 7.1 7.2 7.6 7.2 7.4 12.a 12.5 12.6 26.2 24.8 25.5 41.7 39.6 40.6 NO SURVIVING PUPS
M = MALE F = FEMALE T = TOTAL (SUM OF PUP WEIGHTS/NUM8ER 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.
418-014:PAGE B-81
JohTOO
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 M FT
DAY 4 M FT
DAY 7 M FT
DAY 14 M FT
MATERNAL DOSAGE GROUP I
0 (VEHICLE) MG/KG/DAY
18910 18911 18913 18915 18928 18927 18939 18940
7.4 7.1 7.3
a a a 17.5 16.4 17.1 33.8 32.1 33.1
6.8 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 8.0 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 19.8 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 = 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. Values were not recorded.
418-014:PAGE B-82
/Jofrtoo
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 1): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS
RATS ASSIGNED TO CROSS -FOSTERING
PUP It
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 B
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 5.6 5.9 5.7 6.0 5.9 5.8 5.0 5.5 5.7 5.4 5.5 5.4 5.0 5.7 5.5 5.6 6.1 5.6 5.5 5.5 5.6 5.5 5.6 5.4 5.5 4.9 5.0 5.3 5.5 5.3 5.2 5.6 5.7 5.6 6.0 5.8 6.2 5.7 5.3 5.6 5.3 5.5 5.3 5.4 5.5 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 #
MATERNAL DOSAGE GROUP I P
0 (VEHICLE) MG/KG/DAY
SUBSET B
POSTPARTUM DAY 1
18901 18904 18906 18912 18917 18919 18922 18923 18931 18932 18935 ' 18938
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 .S 6.1 5.6 6.0 5.5 S .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
to vto o
PROTOCOL 4X8-014: 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 #
RAT/ LITTER #
12
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
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 5.7 6.2 6.7 6.2 5.6 6.1 5.8 5.8 5.9 5.8 5.6 5.7 5.5 5.5 5.2 5.5 5.4 5.2 5.7 5.3 5.2 5.6 5.4 5.4 5.2 5.3 5.7 5.5 4.7 6.0 6.4 6.2 6.3 6.0 5.9 6.0 5.9 6.1 5.8 5.6 6.5 5.8 5.8 6.1 6.2 6.2 5.8 5.9 5.9 5.9 5.8 5.5 5.5 5.7 5.5 5.6 5.8 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 5.7 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 tt
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 7.2 6.6 6.8 6.7 7.0 7.3 6.6 6.6 6.4 6.6 6.1 6.2 6.4 6.7 d d 6.5 6.7 7.0 7.0 7.9 7.3 7.3 7.1 6.3 7.0 6.4 6.6 6.0 6.4 5.7 6.2 6.1 6.6 8.4 7.7 7.2 7.8 7.9 7.8 7.3 7.9 7.4 7.6 7.6 7.8 7.0 7.7 5.6 6.3 6.6 6.4 6.5 6.8 6.3 6.4 5.7 6.1 6.1 6.1 5.7 5.9 6.2 5.8 5.8 6.9 7.1 6.7 7.3 7.2 6.9 6.7 7.3 6.9 6.1 7.1 6.8 6.4 6.1 6.3 6.6 7.1 7.4 6.8 7.1 7.3 7.2 6.3 5.5 6.4 6.8 6.7 6.5 7.2 6.0 6.1 6.2 6.0 6.0 6.4 6.0 5.9 5.8 6.0 5.5 5.3 5.9 5.8 5.5 6.1 4.1 6.1 6.2 6.9 6.1 6.2 6.5 5.8 6.1 6.4 6.3 6.3 6.4 6.2 5.4 5.5 6.0 6.1 5.6 5.6 5.7 5.9 6.0 5.8 5.5 6.0 5.7 5.6 5.2 5.8 5.5 5.6 5.5 5.0 5.4 4.9 6.4 6.0 6.1 5.9 5.8 5.9 5.5 5.9 6.1 5.4 5.2 5.3 6.0 5.9 5.8 5.9 ' 6.0 6.4 6.1 6.2 6.4 6.4 6.7 6.3 5.7 6.2 5.9 6.1 5.7 5.4 5.6 5.9 5.6 6.2 6.0 6.1 5.9 6.0 6.3 6.3 6.5 5.8 5.9 6.2 5.9 5.4 5.7 6.0 6.1 5.9 5.9 6.7 7.1 6.1 6.5 6.4 7.0 7.0 6.1 5.9 6.3 6.1 FD 1
ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST LETTER -- M-MALE, F-FEMALE SECOND 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. d. Value was not recorded.
418-014:PAGE B-84
O flJ a
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 n
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 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 8.0 6.7 8.8 7.3 8.4 7.7 8.1 5.3 8.2 7.4 6.7 8.1 FD 4 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 10.5 9.2 9.7 9.5 10.6 10.1 10.4 MM 2 MM 2 9.7 9.0 9.3 9.2 9.0 9.5 9.0 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
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
B .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 8.6 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 group litters. b. Litters were not cross-fostered; values excluded from group averages. c. Vehicle control group dams cross -fostered with vehicle control litters.
0 0 TOO
418-014:PAGE B-85
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.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 It
12
3 4 5 6 18
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 PRECULLING
18943 1B947 18948 18957 18960 18964 18967 18968 18969 18970 18973 1B974
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
a .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 It
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 6.0 4.3 MD 4 MD 4 MM 3 MD 2 MD 2 MD 2 5.2 FM 3 FD 2 FD 2 FD 2 FD 2 FD 2 FD 2 7.3 7.9 7.5 8.3 8.7 7.8 8.3 9.1 6.7 7.2 6.7 8.8 7.4 7.4 7.3 6.3 7.8 7.4 B .B 8.9 8.0 8.6 8.9 9.6 8.8 8.6 8.9 8.3 8.1 7.5 8.7 8.6 8.7 6.3 8.9 8.5 8.3 7.8 8.9 8.4 7.1 6.6 7.8 7.5 7.6 7.7 7.7 7.4 7.4 8.0 8.7 5.5 8.8 8.3 8.4 9.1 8.9 8.8 7.4 8.0 9.0 7.8 7.8 8.3 8.9 9.3 7.8 10.0 8.9 9.5 9.5 7.8 8.1 8.7 8.4 8.8 6.9 8.5 7.7 B .4 8.0 8.0 8.1 7.8 8.5 8.0 8.3 8.8 8.0 7.3 7.3 8.4 7.8 7.0 FM 3 7.8 8.9 8.4 8.0 8.5 6.0 9.1 7.1 6.8 8.5 7.6 7.5 8.1 8.3 8.3 7.7 7.1 7.0 6.8 7.4 7.0 7.3 6.5 6.7 8.0 7.1 7.3 6.5 5.6 6.4 6.9 6.8 6.1 6.6 6.2 8.2 8.5 8.4 7.3 8.1 8.0 7.5 8.2 B .2 7.4 7.3 6.3 7.2 7.2 7.6 8.0 8.6 8.6 8.5 9.7 9.1 8.7 9.1 9.1 8.3 8.5 7.4 8.1 8.2 7.4 7.2 8.6 9.0 8.0 8.0 6.8 7.0 7.7 7.3 7.3 MD 2 7.6 6.7 6.6 6.4 7.0 7.4 7.4 7.4 7.0 7.2 MD 3 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FD 2 FD 2 FD 2 FD 2 FD 1
ALL WEIGHTS WERE RECORDED IN GRAMS (G). FIRST LETTER -- M-MALE, F-FEMALE SECOND LETTER -- A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED) NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. 1.6 mg/kg/day dosage group dams cross-fostered with 1.6 mg/kg/day dosage group litters. b. 1.6 mg/kg/day dosage group dams cross-fostered with vehicle control group litters. c. Litters were not cross -fostered; values excluded from group averages.
20
418-014:PAGE B-86
00141#
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 5) : PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - 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 a
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 7.5 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 .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
FC 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 t)
MATERNAL DOSAGE GROUP I p
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 6.5 7.1 9.0 0.7 1..6 8.3 .0.8 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
Yrt-Too
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 20
RAT/ LITTER (t
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 8.8 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 41'; 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
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 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 4; MM 3 8 .T: 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 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 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-88
001413
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 n
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 16908 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 il.a 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
12.6
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
o
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-89
PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.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 17 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 16966 18969 18970 18973 18974
14.1 12.2
6.0 14.1
9.9 11.0 14.8 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 .3 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.S 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 #
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..1 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., 1 12. 2 FC 4 12 .5 FC 4 12 .3 FC 4 11 .7 FC 4 FC 4 FC 4 FD i
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 FD 2 FC 4 11.2
12.4 FC 4 11.5
FM 3 FC 4 12.5 1H .7 12.0 FC 4 12.8 FC 4 13.1
FC 4
10.9 12.8
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. 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-90
001415"
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 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 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
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 2B .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
o ALL WEIGHTS WERE RECORDED IN GRAMS (G).
o FIRST LETTER -- M-MALE, F-FEMALE
P
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.
vi* 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
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 K 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 IB 19 20
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
RAT/ LITTER #
MATERNAL DOSAGE GROUP 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 20.7 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 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-92
PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.13) TABLE 30 (PAGE 11): 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 it
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
RAT/ LITTER It
MATERNAL DOSAGE GROUP Ip.
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
40.9 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 47.2 47.3 51.8 51.1 45.3 FC 4 50.2 51.4 49.5, MC 4 44.4 42.8 45.2 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 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-93 001418
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 #
12
3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
RAT/ LITTER #
MATERNAL DOSAGE GROUP II a
1.6 MG/KG/DAY
SUBSET C
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 3.3 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 .B 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 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.
TrXOO
418-014: PAGE B-94
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 # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19
RAT/ LITTER #
MATERNAL DOSAGE GROUP i
0 (VEHICLE) MG/KG/DAY
POSTPARTUM1 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.B 6.4 6.3 6.8 6.2 6.4 6.3 6.6 6.2 6.8 6 .a 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 5.4 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 It
MATERNAL DOSAGE GROUP 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 LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED.
20
418-014: PAGE B-95
a -3
Sin
PROTOCOL 418 014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 141: 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 11 12 13 14 15 16 17 18 19
RAT/ LITTER #
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 MD 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
6.4 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 tt
18956 18971
MATERNAL DOSAGE GROUP II
8.8 9.3 9.2 9.6 9.6 MS 9.1 FS
MS
1 .6 MG/KG/DAY
POSTPARTUM DAY 4
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
i
0 Z T O O
PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 15): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS
RATS ASSIGNED TO PHARAMACOKINETIC EVALUATION
PUP tf
1
2
3
4
5
6
7
8
9 10 11 12 13 14 15 16 17 18
RAT/ LITTER #
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 H
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
0 0 1 4 2 JU
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 30 (PAGE 16): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - FI GENERATION PUPS
RATS ASSIGNED TO PHARAMACOKINETIC EVALUATION
PUP tt
1
2
3
4
5
6
7
8
9 10 11 12 13 14 15 16 17 18
RAT/ LITTER ft
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 FM10 18.8 19.0 20.9
21.7 FS
FM 4
17.6
RAT/ LITTER tt
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
f^ T O O
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 # 1 2 3 4 5 6 7 a 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23
RAT/ LITTER tt
MATERNAL DOSAGE GROUP I a
0 (VEHICLE) MG/KG/DAY
SUBSET A
18903 MC 4 M A M A M A M A M A MM 3 F A FC 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 )t
MATERNAL DOSAGE GROUP I 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.
fe frT O d
418-014: PAGE B-99
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 #
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 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 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 18959 MC 4 M A M A M A M A MC 4 M A MC 4 MC 4 F A FC 4 F A F A F A F A FM 3 18961 M A M A M A M A M A FC 4 F A FC 4 FC 4 F A FC 4 FC 4 F A F A F A FC 4 18963 MC 4 M A M A M A MC 4 M A MC 4 MC 4 M A MC 4 MC 4 F A FC 4 FC 4 F A F A F A F A FC 4 18965 M A M A M A MC 4 M A M A F A F A FC 4 FC 4 FC 4 FC 4 F A FC 4 F A F A FC 4 18972 M A MC 4 M A MC 4 M A M A M A F A FC 4 F A F A FC 4 FC 4 FC 4 F A F A 18976 M A M A M A MC 4 M A MC 4 M A MD 2 F A F A FC 4 FC 4 FC 4 FC 4 F A FC 4 F A F A 18977c MD 3 MD 2 MD 2 MD 2 MD 2 MD 2 MD 2 FD 2 FD 2 FD 2 FD 2 FD i
FIRST LETTER -- M-MALE. F-FEMALE SECOND LETTER -- A-ALIVE, S-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-100
S z s TOO
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 S 6 7 B 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23
RAT/ LITTER #
MATERNAL DOSAGE GROUP I
0 (VEHICLE) MG/KG/DAY
18910 18911 , 18913 18915 18926 18927 18939 18940
MA MA MA 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 MA MA MA FA FA FA FA FA FA FA FA FA FA FA FS FS MA MA MA MA MA MA FA FA FA FA 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 M A MA MA FA FA FA FA FA FA FA FA MA MA MA MA MA MA MA MA MA MA FA FA FA FA FA FA
RAT/ LITTER #
MATERNAL DOSAGE GROUP II
L.6 MG/KG/DAY
18956 M A M A M A M A M A M s F A F A F A F A F A F A F A 18971 M S F A F S
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
$ f l/T O O
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 #
DAY(S) POSTPARTUM
OBSERVATIONS a
A Ib 0 (VEHICLE)
18920
3 1/14 PUPS: NOT NURSING.
18941
4 1/9 PUPS: NOT NURSING.
B IC 0 (VEHICLE)
18932
7-10 11-21
1/10 PUPS: TIP OF TAIL, BLACK. 1/10 PUPS: 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
18974
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
o
N
PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.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
DAY(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
6 1/17 PUPS: NOT NURSING.
a. Tabulation restricted to adverse observations; all other pups appeared normal.
fczfrTOO
418-014: PAGE B-103
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
DAY(S) POSTPARTUM
OBSERVATIONS a
I 0 (VEHICLE)
18904 18912
1 1
3 PUPS: STILLBORN. 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL.
18923 18937
1 1
1 PUP: FOUND DEAD. 1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL.
II
1. e
18952
1
1 PUP: STILLBORN. ALL TISSUES APPEARED NORMAL.
18973
1
1 PUP: FOUND DEAD. NO MILK IN STOMACH. BACK: RAISED AREA (2.0 CM X 1.3 CM X 0.3 CM). CONTAINED GAS.
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
z Y lO O
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 #
DAY(S) POSTPARTUM
OBSERVATIONS a
A Ib 0 (VEHICLE)
18903 18907
4,21 4
21
ALL PUPS APPEARED NORMAL.
1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL.
ALL PUPS APPEARED NORMAL.
18908
4
1 PUP: FOUND DEAD. AUTOLYSIS PRECLUDED FURTHER EVALUATION.
18909 18918
18920
4 21
2
4
ALL PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL.
1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL.
ALL PUPS APPEARED NORMAL.
18924 18934
4 21
2 21
1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL.
ALL PUPS APPEARED NORMAL.
1 PUP: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER TISSUES APPEARED NORMAL.
ALL PUPS APPEARED NORMAL.
18936
21
ALL PUPS APPEARED NORMAL.
18941 18942
2 4
2 PUPS: FOUND DEAD. ALL TISSUES 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 1.6 mg/kg/day dosage group litters.
418-014:PAGE B-105
V oo
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 (t
DAY (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.
PROTOCOL 418-014: ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.13) TABLE 33 (PAGE 4): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - FI GENERATION PUPS
RATS ASSIGNED TO CROSS -FOSTERING
SUBSET MATERNAL DOSAGE GROUP MATERNAL DOSAGE (MG/KG/DAY)
FOSTER LITTER #
DAY(S) POSTPARTUM
OBSERVATIONS a
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. ALL 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 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 1.6 mg/kg/day dosage group litters.
418-014: PAGE B-107
'TCfrTOO
PROTOCOL 418-014: ORAL (GAVAGE) CROSS -FOSTER ING 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 ft
DAY (S) POSTPARTUM
OBSERVATIONS a
D II b 1.6
18946
2 5
2 PUPS: FOUND DEAD. ALL TISSUES APPEAREDI NORMAL. 7 PUPS: FOUND DEAD. NO MILK IN STOMACH. ALL OTHER
TISSUES APPEARED NORMAL. 1 PUP: !FOUND DEAD. ALL TISSUES APPEARED NORMAL.
18951 18952 18953 18958 18961 18963 18965 18972
18976
21 21 4,21 4,21 4,21
4 4 21
2
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.
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
f frT O O
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
DAY(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.
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
oo
h*
APPENDIX C PROTOCOL AND AMENDMENT
00143^
418-014: PAGE C-1
O P r im e d ic a
Argus Research Laboratories, Inc. 905 Sheehy Drive, Building A Horsham, PA 19044 Telephone: (215) 443-8710 Telefax: (215) 443-8587
PROTOCOL 418-014
SPONSOR'S STUDY NUMBER: T-6295.13
STUDY TITLE:
Oral (Gavage) Cross-Fostering Study of PFOS in Rats
PURPOSE:
The purpose of this study is to evaluate pup survival of F1 rats following PFOS treatment of Crl: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
00143^
418-014: PAGE C-2
Protocol 418-014 Page 2
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.
00143JT
418-014: PAGE C-3
Protocol 418-014 Page 3
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:
Roomtemperature. Roomtemperature. Roomtemperature. Room temperature.
00143$
418-014: PAGE C-4
Protocol 418-014 Page 4
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 wer prepared.
418-014:PAGE C-5
Protocol 418-014 Page 5
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 Cri: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 Facility0'35; and 3) the test article is pharmacologically active in the species and strain.
G 0 1 4 f&
418-014:PAGE C-6
Protocol 418-014 Page 6
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 Aoe:
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.
00144
418-014: PAGE C-7
Protocol 418-014 Page 7
ANIMAL HUSBANDRY:
All cage sizes and housing conditions are in compliance with the Guide for the Care and Use of Laboratory Animals^.
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 rat's cage. Beginning no later than day 20 of presumed gestation, Fo generation female rats will be individually housed in nesting boxes, except durirjg 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.
00144JL
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Protocol 418-014 Page 8
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-fostering
418-014: PAGE C-9
Protocol 418-014 Page 9
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) Utters 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
D
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.
418-014: PAGE C-10
Protocol 418-014 Page 10
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 Pups:
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-4184)14-A(Day.Month.Year) 5 B-418-014-B(0ay.Month.Year)
The test article will be considered 100% pure for the 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.
418-014:PAGE C-11
Protocol 418-014 Page 11
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.
418-014:PAGE C-12
Protocol 418-014 Page 12
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 (-70 C or below) until shipment to the Sponsor for analysis.
418-014: PAGE C-13
Protocol 418-014 Page 13
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 (-70C 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.
00144J
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Protocol 418-014 Page 14
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 (-70C or below) until shipment to the Sponsor for analysis.
00144
418-014: PAGE C-15
Protocol 418-014 P age 15
Dams with No Surviving Pups:
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.
418-014.-PAGE C-16
Protocol 418-014 Page 16
Clinical Observations and/or Generai 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.
001452
418-014: PAGE C-17
Protocol 418-014 Page 17
Puds 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 - Days 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 Dav 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.
0014L
418-014:PAGE C-18
Protocol 418-014 Page 18
The pups will be examined for gross lesions. The liver from each pup will be collected, pooled (per litter), frozen and stored (-70 C 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. AH 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.
0014
418-014:PAGE C-19
Protocol 418-014 Page 19
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 o f 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
001454
418-014: PAGE C-20
Protocol 418-014 Page 20
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.
001455"
418-014: PAGE C-21
Protocol 418-014 Page 21
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 CrUCDBR 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. Saiewski, E. (1964). Farbemethode zum makroskopischen Nachweis von Implantationsstellen am Uterus der Ratte. Arch. Pathol. Exp. Pharmakol. 247:367.
00145$
PROTOCOL APPROVAL: FOR THE TESTING FACILITY
418-014:PAGE C-22
Protocol 418-014 Page 22
a t -i* 1 --
-- ________
Alan M. Hoberman, Ph.D., DABT Director of Research
Date
Study Director
Zi-Oc-T- 9 3 Date
~ rDena C. Lebo, V.M.D. Member, Institutional Animal Care and
Use Committee
FOR THE SPONSOR
Date
9?
Marvin T. Case, D.V.M., Ph.D. Study Monitor
Date
00145JT
418-014: PAGE C-23 ATTACHMENT 1 SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE
00145$
ATTACHMENT 1
418-014:PAGE C-24
Protocol 418-014 Page 1 of 2
STUDY SCHEMATIC CROSS-FOSTERING STUDY"
Startof End of Dosage Dosage
Scheduled Sacrifice^
Premaing Period (42 Days)
Cohabitation Period (5 days)
Presumed Gestation Period
Postpartum/ Lactation Period
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.
_,, ,,_^
00145J
418-014: PAGE C-25
ATTACHMENT 1
Protocol 418-014 Page 2 of 2
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.
418-014:PAGE C-26 ATTACHMENT 2 MATERIAL SAFETY DATA SHEET
00146#
418-014: PAGE C-27
MATERIAL SAFETY DATA SHEET
3M 3M Cantar St. Paul, Minnesota 55144-1000
1 -BOO-364-3577 or (612) 737-6501 (24 hours)
Copyright, 1998, Minnesota Mining and Manufacturing Coapany. All rights reserved. Copying and/or downloading of this inforaation for the purpose of properly utilizing 3M products is allowed provided that: 1) the inforaation is copied in full with no changes unless
prior agreeaent is obtained froe 3M, and 2) neither the copy nor the original is resold or otherwise
distributed with the intention of earning a profit thereon.
DIVISION: 3M CHEMICALS TRADE NAME:
FC-95 FLU0RAD Brand Fluorocheaical Surfactant ID NUMBER/U.P.C.:
98-0207-0103-7 00-51135-090S4-1 98-0207-0104-5 98-0211-0888-5 00-51135-09362-7 98-0211-3916-1 ZF-0002-1044-1 ISSUED: January 29, 1998 SUPERSEDES: Noveaber 05, 1997 DOCUMENT: 10-3796-9
00-51135-09055-B 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 -8 -7 -6 -3
2. PHYSICAL DATA
BOILING POINT:..... VAPOR PRESSURE:.... VAPOR DENSITY:..... EVAPORATION RATE:... SOLUBILITY IN WATER: SPECIFIC GRAVITY:...
PERCENT VOLATILE:...
P H : .............................................
VISCOSITY:.......... MELTING POINT:.....
N/A N/A N/A N/A slight ca. 0.6 Water~1
(Bulk)
0%
7 -B (0.1% Aqueous)
N/D N/D
APPEARANCE AND ODOR: Light colored, free flowing powder.
A b b r e v i a t i o n s : N /D - N o t D e t e r m in e d N /A - N o t A p p l i c a b l e CA - A p p r o x im a t e ly
418-014:PAGE C-28
USDS: PC-86 FLUORAD Brand Fluorocheaical Surfactant
January 29, 1996
PAGE 2
3. FIRE ANO EXPLOSION HAZARD DATA
FLASH POINT:....................Nona
FLAMMABLE LIMITS -
LEL:.. N/A
FLAMMABLE LIMITS -
UEL:....N/A
AUTOIGNXTION TEMPERATURE:..... N/A
EXTINGUISHING MEDIA: Mat or, Carbon dioxide, Dry chemical, Foaa
SPECIAL FIRE FIGHTING PROCEDURES: Hear full protective clothinoi including helmet, self-contained, positive pressure or pressure deaand breathing apparatus, bunker coat and pants, bands around aras, waist and legs, face aask, and protective covering for exposed areas of the head.
UNUSUAL FIRE AND EXPLOSION HAZARDS: See Hazardous Decoaposition section for products of coabustion.
4. REACTIVITY OATA
STABILITY: Stable
INCOMPATIBILITY - MATERIALS/CONDITIONS TO AVOID: Not applicable.
HAZARDOUS POLYMERIZATION: Hazardous polyaerization will not occur.
HAZARDOUS DECOMPOSITION PRODUCTS: Carbon Monoxide and Carbon Dioxide, Oxides of Sulfur, Hydrogen Fluoride, Toxic Vapors, Gases or Particulates.
5. ENVIRONMENTAL INFORMATION
SPILL RESPONSE: Observe precautions fros othar sections. Vacuus, use wet sweeping compound or water to avoid dusting. CAUTION! A vacuua cleaner could be an ignition source. Clean up residue with water. Place in an approved metal container. Seal the container.
RECOMMENDED DISPOSAL: Do not release to waterways or sewer. Oo not use in products or processes that could result in aquatic concentrations greater than 1/10 of the lowest ECSO or LC50 concentration. Incinerate in an industrial or coaaercial facility in the presence of a coabustible material. Combustion products will Include HF. Disposal alternative: Dispose of waste product in a facility permitted to
00146S
A b b r e v i a t i o n s : N /D - N o t D e t e r m in e d N /A - N o t A p p l i c a b l e CA - A p p r o x im a t e ly
418-014: PAGE C-29
USDS: FC-95 PLUOBAO Brand Fluorochemical Surfactant January 29, 1998
PAGE 3
5. ENVIRONMENTAL INFORMATION
(continuad)
accept cheaical Hast.
ENVIRONMENTAL DATA: 96-Hr. Aquatic Fish LCSO, Fathead Minnow(Pimephales proaelas)-38 mg/1, Bluegill Sunfish (Lepoais macrochirus)68 ag/1, Rainbow Trout(Salso gairdneri)1l ag/1,' 48-Hr. EC50, Daphnia Magna * 50 ag/1; COO.004 g/g; 80020 Nil.
REGULATORY INFORMATION: Volatile Organic Coapounda: N/A. VOC Less H20 & Exeapt Solvents: N/A.
Since regulations vary, consult applicable regulations or authorities before disposal. U.S. EPA Hazardous Haste Nuaber None (Not U.S. EPA Hazardous).
This product complies with the cheaical registration requireaents of TSGA, EINECS, COSL, AICS, MITI and Korea.
EPCRA HAZARD CLASS: FIRE HAZARD: No PRESSURE: No REACTIVITY: No ACUTE: Yes CHRONIC: Yes
6. SUGGESTED FIRST AID
EYE CONTACT: Immediately flush eyes with large amounts of water for at least 15 minutes. Get immediate medical attention.
SKIN CONTACT: Immediately flush skin with large amounts of water. Remove contaminated clothing. If irritation persists, call a physician. Wash contaminated clothing before reuse.
INHALATION: If signs/symptoms occur, reaove person to fresh air. If signs/symptoms continue, call a physician.
IF SWALLOWED: Drink two glasses of water. Call a physician.
7. PRECAUTIONARY INFORMATION
EYE PROTECTION: Avoid eye contact. Wear vented goggles.
0014G
A b b r e v i a t i o n s : N /D - N o t D e t e r m in e d N /A N o t A p p l i c a b l e CA - A p p r o x im a t e ly
418-014: PAGE C-30
HSDS: FC-Q5 FLUORAD Brand Fluorochemical Surfactant
January 29, 1998
7. PRECAUTIONARY INFORMATION
(continued)
PAGE 4
SKIN PROTECTION: Avoid akin contact. Near appropriate gloves when handling this Material. A pair of gloves sade from the following aaterial(s) are recoaeendad: butyl rubber. Use one or sore of the following personal protection itens as necessary to prevent skin contact: head covering, coveralls. Protective gareents (other than gloves) should be nade of either of the following Materials: polyethylene/polyvinylidene chloride (Saranex).
RECOMMENDED VENTILATION: Use with appropriate local exhaust ventilation. Use in a wellventilated area. Provide sufficient ventilation to Maintain emissions below reconnended exposure liaits. If exhaust ventilation is not adequate, use appropriate respiratory protection.
RESPIRATORY PROTECTION: Avoid breathing of dust. Select one of the following NIOSH approved respirators based on airborne concentration of contaainants and in accordance with OSHA regulations: half-aask dust and Mist respirator, half-eask supplied air respirator, full-face dust and aist respirator, full-face supplied air respirator.
PREVENTION OF ACCIDENTAL INGESTION: Do not eat, drink or saoke when using this product. Nash exposed areas thoroughly with soap and water. Nash hands after handling and before eating.
RECOMMENDED STORAGE: Keep container dry. Keep container closed when not in use.
FIRE AND EXPLOSION AVOIDANCE: Nonflammable.
OTHER PRECAUTIONARY INFORMATION; No snoking: Smoking while using this product can result in contamination of the tobacco and/or saoke and lead to the formation of the hazardous decomposition products eentioned in section 4 of this HSDS.
HMIS HAZARD RATINGS: HEALTH: 2 FLAMMABILITY: 0 REACTIVITY: 0 PERSONAL PROTECTION: X (See precautions, section 7.)
EXPOSURE LIMITS
INGREDIENT
POTASSIUM PERFLUOROALKYL SULFONATE... POTASSIUM PERFLUOROALKYL SULFONATE... POTASSIUM PERFLUOROALKYL SULFONATE... POTASSIUM PERFLUOROALKYL SULFONATE...
VALUE UNIT
0.1 MG/M3 0.1 MG/M3 0.1 MG/M3 0.1 MG/M3
TYPE
TWA TWA TWA TWA
AUTH SKIN
3M 3M
Y Y
00146^
3M Y
3M Y
A b b r e v i a t i o n s : N /D - N o t D e t e r m in e d N /A - N o t A p p l i c a b l e CA - A p p r o x im a t e ly
418-014:PAGE C-31
JHaSnOuSa:ryFC2-99,5 F1L9U9O8RADBrand Fluorochaaical Surfactant
EXPOSURE LIMITS
(continuad)
PAGE 5
INGREDIENT
VALUE UNIT
TYPE AUTW SKIN*
POTASSIUM PERFLUOROALKYL SULFONATE... 0.1 MG/M3
TWA SM
Y
- SKIN NOTATION: Listed substances indicated with *Y' under SKIN refer to the potential contribution to the overall exposure by the cutaneous route including aucous aeabrane and eye, either by airborne or, aore particularly, by direct contact Mith the substance. Vehicles can alter skin absorption.
SOURCE OF EXPOSURE LIMIT DATA: - 3M: 3M Recoaaended Exposure Guidelines
8. HEALTH HAZARD DATA
EYE CONTACT: Mild Eye Irritation: signs/syaptoas can include redness, swelling, pain, and tearing.
SKIN CONTACT: Mild Skin Irritation (after prolonged or repeated contact): signs/syaptoas can include redness, swelling, and itching.
Hay be absorbed through the skin and persist in the body for an extended tiae.
INHALATION: May be haraful if inhaled.
May be absorbed by inhalation and persist in the body for an extended time.
Single overexposure, above recoaaended guidelines, aay cause:
Irritation (upper respiratory): signs/syaptoas can include soreness of the nose and throat, coughing and sneezing.
IF SWALLOWED: Ingestion is not a likely route of exposure to this product.
Illness aay result froa a single swallowing of a moderate quantity of this eaterial.
May be harmful if swallowed.
MUTAGENICITY: Mutagenicity assays indicate the product is not mutagenic.
00146
A b b r e v ia t io n s : N /D - N o t D e t e r a in e d N /A - N o t A p p lic a b le CA - A p p r o x im a te ly
418-014: PAGE C-32
HSDS: FC-95 FLUORADBrand Fluorocheaical Surfactant
January 29, 1998
PAGE 6
B. HEALTH HAZARD DATA
(continued)
REPRODUCTIVE/DEVELOPMENTAL TOXINS: Not teratogenic in the rat at oral doses below natemally toxic levels.
OTHER HEALTH HAZARD INFORMATION: This product is not known to contain any substances regulated under California Proposition 65.
A Product Toxicity Suaaary Sheet is available.
SECTION CHANGE OATES
HEADING
SECTION CHANGED SINCE Noveaber OS, 1997 ISSUE
Abbreviations: N/D - Not Oeterained N/A - Not Applicable CA Approximately
The inforeation in this Material Safety Data Sheet (MSDS) is believed to be correct as of the date issued. 3M MAKES NO WARRANTIES, EXPRESSED OR IMPLIED, INCLUDING, BUT NOT LIMITED TO, ANY IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE OR COURSE OF PERFORMANCE OR USAGE OF TRADE. User is responsible for deteraining whether the 3M product is fit for a particular purpose and suitable for user's sethod of use or application. Given the variety of factors that can affect the use and application of a 3H product, soae of which are uniquely within the user's knowledge and control, it is essential that the user evaluate the 3H product to deteraine whether it is fit for a particular purpose and suitable for user's aethod of use or application.
3M provides information in electronic fora as a service to its custoaers. Due to the remote possibility that electronic transfer aay have resulted in errors, oaissions or alterations in this inforaation, 3M makes no representations as to its completeness or accuracy. In addition, information obtained from a database may not be as current as the information in the MSDS available directly froa 3M.
00146
418-014: PAGE C-33 ATTACHMENT 3 TEST ARTICLE PREPARATION PROCEDURE
00146$
418-014:PAGE C-34
ATTACHMENT 3
Protocol 418-014
Version: 4 18 -0 14 (22 O C T 98)
TEST ARTICLE PREPARATION PROCEDURE
Page 1 of 2
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
3. Vehicle will be prepared:
___ Daily
_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.
00146
418-014:PAGE C-35
ATTACHMENT 3
Protocol 418-014
Version: 4 1 8 -0 1 4 (22 O C T 981
TEST ARTICLE PREPARATION PROCEDURE
Page 2 of 2
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 :
OO14JT0
418-014:PAGE C-36 ATTACHMENT 4 CROSS-FOSTERING PROCEDURE
00147#
418-014: PAGE C-37
ATTACHMENT4
CROSS-FOSTERING PROCEDURE
Protocol 418-014 Page 1 of 1
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.
00147JL,
418-014: PAGE C-38
O P rimedica
Arg9u0s5TReSelThseeepeealherhoHcfyanhoxeDrL::srah((i2bv2aoe1m1,r55a,)B)t4Po4u4A4rii3l3ed--1si88n,957g0I18n4A0c74.
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.
00147J
418-014: PAGE C-39
Protocol 418-014 Amendment 1 Page 2
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-Fosterina 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.
Alan M. Hoberman, Ph.D., DABT Director of Research
IW ) Date
Dnd G. York, Ph.D., DABT Date Associate Director of~F?esearch and
Study Director
Dena C. Lebo, V.M.D.
Date
Chairperson, Institutional Animal Care and
Use Committee
MarvinT. Case, D.V.M., Ph.D. Study Monitor
Date
00147.
418-014:PAGE C-40
Protocol 418-014 Version: 4 1 8 -0 1 4 (2 9 Dec 96)
Page 1 of 2
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
x 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 in protocol.
7. Storage: Room temperature
001475'
418-014: PAGE C-41
Protocol 418-014 Version: 4 1 8 -0 1 4 (29 Dec 981
Page 2 of 2
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 11,6g of SodiumPhosphateMonobasic to the container and stir until uniform.
5. Add 2.70g of SodiumHydroxide to the vessel and stir until uniform.
[NOTE: The final solution should have a pH of approximately 7.3 ]
Written By:
-V-
Approved b
Date:
Clarificationx: _yy.No _Yes [see attached clarification form]
Initials/Date:
'of?*}
00147$
APPENDIX D DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING
PROCEDURES OF THE TESTING FACILITY
00147JT
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.
r' >n -*L i
APPENDIX E TEMPERATURE AND RELATIVE HUMIDITY REPORTS
\H-7fQ Q 1 1W u ZjKt j T^w J|
418-014:PAGE E-1
ARGUS
TemperatureLaoncadtiRonel:aRtivoeomHu0m4idity Report Protocol Number: 418-014
Range of Dates: 27-Oct-1998 13:15to 23-Nov-1998 15:49
Target Range: Species: Rat
Total Number of Days: Total Number of Hours: Total Number of Data Points:
Temperature 64*F to 79F
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: -ff 7?
Cumulative by Location (v04.01.97)
o o 1? (flfO
418-014: PAGE E-2
ARGUS
TemperatureLoancadtiRoenl:aRtivoeomHu2m7idity Report 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:________ % . 'lUjlL_________________ DATE: /-g ??
Cumulative bv Location fv04.01.9T>
0 0 4 :4 0
\W !
418-014:PAGE E-3
ARGUS
TemperatuLreocaantdionR:elRaotiovme H35u-m3i7dity Report 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:
Mean ( SD):
Maximum: Median: Minimum:
Number of Points in Range (%): Number of Points High (%): Number of Points Low (%):
Temperature 64'F to 79*F
Relative Humidity 30% to 70%
19 426.24
427
19 426.24
427
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 BY:_____________% . /Mm ZL____________DATE: f
Cumulative by Location fv04.01.97)
0014S|^
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 79F
12 265.0 266
Relative Humidity 30% to 70%
12 265.0 266
Mean ( 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:
/th./nxL-
DATE: i-f-1 1
Cumulative bv Location Cv04.01.971
00145^,
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-l 998 14:00
R.H.
28.6 L
30% to 70%
Date
Time
R.H.
H = Value out of range - High L = Value out of range - Low R.H. = Relative Humidity (%)
Report Generated: 13-May-1999 at 10:45
These deviations did not adversely affect the outcome or interpretation of the study. _____The following deviation^) impacted on the outcome of the study as described:
Deviations by Location (v04.01.97)
OOl
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.
5.8 L 5.1 L 5.0 L 5.1 L 5.1 L
5.2 L 5.3 L 5.5 L 5.6 L 5.6 L
30% to 70%
Date
Time
H = Value out of range - High L = Value out of range - Low R.H. = Relative Humidity (%)
Report Generated: 13-May-1999 at 10:50
These deviations did not adversely affect the outcome or interpretation of the study. The following deviation(s) impacted on the outcome of the study as described:
Deviations bv Location (v04.01.97)
418-014:PAGE E-7
ARGUS
Relative Humidity Deviations Report Location: Room 27
Protocol Number: 4184)14
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.
70.4 H
30% to 70%
Date
Time
H = Value out of Fange - High L = Value out of range - Low R.H. = Relative Humidity (%)
Report Generated: 13-May-1999 at 10:54
These deviations did not adversely affect the outcome or interpretation of the study. The following deviations) impacted on the outcome of the study as described:
Deviations by Location (v04.01.97)
0014$^
APPENDIX F PATHOLOGY REPORT
00148^
418-014:PAGE F-1
Pathology Associates International
A Company of Science Applications International Corporation
A n tm otoy+-O w r>+<3 C o r n e r
PATHOLOGY REPORT (ANCILLARY STUDY)
ELECTRON MICROSCOPIC EVALUATION OF LIVER AND LUNG IN CRL:CDBR VAF/PLUS RATS)
ORAL (GAVAGE) CROSS-FOSTERING STUDY OF PFOS IN RATS
CLIENT STUDY NUMBER 418-014 PAI STUDY NUMBER EM 99.46
Prepared by: James B. Nold, D.V.M., Ph.D., Diplomate, A.C.V.P.
Pathology Associates International 4915-D Prospectus Drive Durham, NC 27713
April 5,1999
PREPARED FOR 3M CORPORATE TOXICOLOGY, 3M CENTER, ST. PAUL, MN 55144-1000
00148$
4915 D Prospectus Drive Durham, North Carolina 27713 (919) 544-5257 (919) 544-3218 FAX
418-014:PAGE F-2
Pathology Narrative Transmission Electron Micrograph Interpretation Forms Transmission Electron Micrographs Quality Assurance Statement
Section I II m IV
00148
418-014:PAGE F-3 L Pathology Narrative
0014fg>
418-014:PAGE F-4
PrimedicaAAncriglluasryStPuadtyhoNloog. y41R8e-0p1o4n
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 increasednumbers of 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 of peroxisomes in treated animals. Differences in cellular membranes or mitochondria were not discernible between control andtreated animals in the samples examined. Type II pneumocytes containing lamellar bodies were identified in both controls andhigh-dose animals; however, there appearedto bean 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 of this study was toexamine by electron microscopy the livers and lungs from selected day-1 postpartumrat 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 of peroxisomes within hepatocytes, membrane andmitochondrial changes in hepatocytes, andthe quantitation of surfactant lamellar bodies in the lung.
For the cross-fostering study, 66 female Crl:CDBR VAF/PLUS rats were given the test article or the vehicle once daily beginning 42 days prior to cohabitation with a male through day 21 postpartumaccording toText Table 1.
Text Table 1. Dosage Levels, Concentrations and Volumes
DGorsoaugpe I
n
NuFmeb2m34e0arl++oe f66R*aMatasted
P(F0mO(gVS/keghD/idoclaseay)g)e
1.6
PFOS (mCogn/mceLn)tration 0.032
Dos(amgeL/Vkogl)ume 5 5
a S1a4mppolsetpsafrotruman.alTyhsiesseofdpahmasrmanadcotkhieniretriecsspwecetrievecoliltlteecrtsedwefrroemsa6crdifaimcesdafnodllothweiinrgressapmecptlievecoliltlteecrtsioonn. day
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The first ten pups from the control pool (irrespective of litter) and the first 10 pups that were found dead (no autolysis present) from the treated group determined to be at day 1 postpartumwere decapitated at approximately 1-3 hours after birth, and the lungs and liver were collected. Samples of lung and liver were thin-sliced andplaced in McDowellTrump fixative (McDowell EM, 1976), and submitted to this laboratory for electron microscopy processing andevaluation.
Per sponsor request, tissue samples fromselected pups were processed and evaluated ultrastructurally (Text Table 2). Pup numbers were assignedby this laboratory because submitted samples were identified by damnumber only. The five digit number is the damnumber, the suffix 1or 2 denotes the pup number from that dam'slitter.
Text Table 2. Animals Selected for Electron Microscopy
1 Control11111C88888O9999910110m24554z-----1112/2kg/day)
High-Dos11e118888(999915755.62362----m1112g/kg/dav)
The lungs and livers were thenprocessedinto Spurr'sresin for ultrastructural examination. Thick (l(i) sections were cut, stained with toluidine blue, and examined to select representative areas for further electron microscopy processing. Thin sections (approximately 90 nm) were cut, mounted on 200-mesh copper grids, stained with 5% methanolic uranyl acetate andReynold'slead citrate, and examined on aZeiss 900 transmission electron microscope. Centrilobular hepatocytes, where clearly identifiable in liver sections, were preferentially examined. Representative electron photomicrographs of liver and lung were taken and significant ultrastructural features were summarized for each photograph and animal on adesignated transmission electron micrograph interpretation form. The number of peroxisomes in hepatocytes was manually counted for each center photographed hepatocyte andrecorded.
RESULTS AND DISCUSSION
Individual interpretations of light andelectron micrographs for each animal are shown in Section II. Selected representative electron photomicrographs were labeled and are shown in Section HI.
Fixation of 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 of the fixative into the tissue. Suboptimal fixation was characterized by discontinuous plasma membranes, lossof detail of cytoplasmic organelles, dilatation of endoplasmic reticulum and the nuclear envelope, and dilatation of mitochondrial cristae. Fixation, however, was still considered adequate to assess for
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the presence of peroxisomes in the hepatocytes and lamellar bodies in the lung and to make general comparisons between control and treated samples.
LIVER
Control (0 mg/kg/day). Subcellular features of control hepatocytes are demonstrated in Figures 1and 2. Hepatocytes characteristically contain numerous mitochondria and prominent endoplasmic redculum. In these cells rough endoplasmic reticulum was evident, but smooth endoplasmic reticulum was not because of suboptimal fixation. Mitochondria generally appeared dark andcontained scattereddense deposits and some dilated cristae, both artifacts due to suboptimal fixation. Dilatation of endoplasmic reticulum and the nuclear envelope were additional artifacts of poor fixation. Fine granular material in the cytoplasmisconsistent with glycogen particles. Peroxisomes were identified and quantitated in fromrepresentative hepatocytes. For the control livers, the average number of 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
Control (0 mg/kg/day)
J1 Anim1111188888a99999l11100N52544--u---11122mber
Numpbeerr HofepPaetroocxvisteomes 46698.....48620
Mean Control =
7.0
High-Dose (1.6 mg/kg/day)
Animal Number 11118888999975553262----1112
Numpbeerr HofepPaetroocxvisteomes 11117674.2...0005
Mean High-Dose =
16.1
High-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
Control (0 mg/kg/day). Lung samples were atelectatic and not inflated with fixative. Therefore most alveoli and alveolar septawere collapsed with poor delineation of alveolar and airway spaces. Figures 6-8 illustrate some normal cellularity and ultrastructural pulmonary anatomy. Type II pneumocytes (PN2) were infrequently identified lining alveoli, terminal bronchioles, or within collapsed septa. Type II pneumocyteswere identified by the presence of lamellar bodies, which are cytoplasmic
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vacuoles containing electron-dense laminated membrane-like profiles. Lamellar bodies represent the material secreted into the alveolar spaces assurfactant. Surfactant or lamellar material within airway spaceswas very rarely identified in control lungs. The type I pneumocyte (PN1) is characteristically flattened and lines the alveolar space asa thin margin of cytoplasm. Type I pneumocytes differentiate fromtype II pneumocytes. Many pneumocytes appeared to becuboidal without distinctive lamellar bodies, and may represent a transition stage between PN2s and PNls, 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 andlamellar bodies were infrequently identified in these control lungs and no reliable quantitation could be assessedof them. Some lamellar-like bodies were also present within capillary lumina.
High-Dose (1.6 mg/kg/day). Ultrastructural pathology of the high-dose lungs was essentially similar to control samples. There appeared, however, to be increased numbers of type II pneumocytes and lamellar bodies; at least, identifying themfor illustrative purposes was easier than in control samples. The collapsed nature of most of the alveoli and septa made definitive quantitation impractical. Some lamellar bodies were identified within capillary lumina andonly asmall number were seen within alveoli. Figures 9-12 illustrate some representative lung fields fromthe high-dose animals.
CONCLUSION
Ultrastructural evaluation of liver andlungs of rat pups fromdams treated with 0 (control) or 1.6 mg/kg/day (high-dose) PFOS identified increased numbers of peroxisomes within hepatocytes of high-dose rat pup livers. Quantitation of 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 of 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 of 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
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REFERENCES McDowell, EM andTrump, BF: Histologic fixative for routine diagnostic light and
electron microscopy. Arch. Pathol. Lab. M ed., 100:405-414, 1976.
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IL Transmission Electron Micrograph Interpretation Forms1
1Forms are typographically corrected versions of signed/datedraw data sheets maintained in archived study file.
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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: RCarlt:CD-BR VAF/Plus
Tissue: Liver_____________
Block No(s).: 99.46-i________ Z15683, Z15684,
Photo/Negative No(s).: Z 15686-88
Significant Lesions (check one): ______ Yes
X No
Interpreting Pathologist (signature anddate): ______________
Features: Z15683:- Hepatocyte, several erythrocytes in sinusoid, and nucleus of probable endothelial cell onright sideof 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 of 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. Z15686: Single hepatocyte surroundedby 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. Z15687: 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. Z15688: 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.___________________________________ _
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TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION
Study No.: 418-014: EM-99.46
Animal No.: 18904-1_______ Treatment Group: 0 mg/kg Control Sex: non applicable______
Species/Strain: RCarlt:CDBR VAF/Plus
Tissue: Lung_____________
Block No(s).: 99.46-2_______ Z15689-
Photo/Negative Nofsl.: Z 15693
Significant Lesions (check one): ______ Yes
X No
Interpreting Pathologist (signature and date): ______________
Features: Z15689: Capillary with three endothelial cell nuclei. In lumen of capillary is a lamellar-like body. Several type 1pneumocytes (PN1) present surrounding capillary. No pneumocyte lamellar bodies present. Z15690: Type 2 pneumocyte (PN2) present, containing multiple lamellar bodies (-7). Beneath the PN2 is a capillary containing an erythrocyte, platelet, andendothelial cell nuclei. PN1 to right side of photograph with more translucent cytoplasm. Z15691: 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 PNls in collapsed septa. No pneumocyte lamellar bodies evident. Z15692: Collapsed alveolar septa. On top is a thin-layered PN1, and in the bottom left is larger profile of a PN1. Centrally appears tobe acapillary lumen containing some lamellar debris and two leucocytes. Also some lipid debris is present in the capillary lumen. Endothelial cell cytoplasmis evident, but no endothelial cell nucleus. No pneumocyte lamellar bodies identified. Z15693: Collapsed alveolar septa. At top is a profile of aPN1in the alveolar space. Beneath it is a thin-layered PN1, and to the left side of photo is a PN1. Subjacent nuclei are probably endothelial cell nuclei. At bottomleft is anerythrocyte. 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.
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TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION
Study No.: 418-014: EM-99.46
Animal No.: 18904-2________ Treatment Group: 0 me/kg/ Control Sex: non applicable______
Species/Strain: CRarltiCDOBR VAF/Plus
Tissue:
Liver___________
Block No(s).: 99.46-3________ Z 15694 -
Photo/Negative No(sl.: Z 15698
Significant Lesions (check one): ______ Yes
X No
Interpreting Pathologist (signature anddate): ______________
Features: Z 15694: Hepatocyte. Multiple mitochondria, some RER and multiple rounded blebs of RER. Cytoplasmfull of granular glycogen. Poor endothelial cell definition in sinusoids. Nucleated erythrocyte in bottomleft. Mitochondria appear dense with some granular deposits anddilated cristae. Three (3) peroxisomes identified. Z15695: Hepatocyte. Dilated RER. Multiple dense mitochondria with granular deposits and dilated cristae. Cytoplasmic margins and endothelial cells poorly defined. Six (6) peroxisomes identified. Z15696: Hepatocyte. Many dilated blebs of RER. Dense mitochondria with granular deposits and dilated cristae. Top left hepatocyte has swollen, less dense, mitochondria. Twelve (12) peroxisomes identified. Z 15697: Several poorly-fixed hepatocytes. Many light and dark dilated blebs of RER, which are difficult to differentiate fromperoxisomes. Five (5) peroxisomes identified in
single more-circumscribed hepatocyte. ZI5698: 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.
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TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION
Study No.: 418-014: EM-99.46
Animal No.: 18904-2_______ Treatment Group: 0 mg/kz Control Sex: non applicable_______
Species/Strain: Crl:CD<B>BR vaf/pius
_EI
Tissue: Lung_____________
Block No(s).: 99.46-4________ 15699 -
Photo/Negative No(s).: Z 15704
Significant Lesions (check one): ______ Yes
X No
Interpreting Pathologist (signature anddate): ______________
Features: Z15699: Alveolus containing several cuboidal pneumocyte cells. The lower cell appears to contain acouple small lamellar bodies, probably aPN2. The other two large cuboidal cells do not contain any lamellar bodies, but appear to bepneumocytes which haven't flattened out into PNls or are terminal bronchiolar epithelia. At the left andbottomare capillaries lined by endothelial cells. The flattened PN1 at the bottom contains asegment of luminal surface with some dense electron-dense material. Some dilated cristae in mitochondria. Z15700: Alveolar space with capillary/erythrocyte at left andpneumocytes around top and right. Top left pneumocyte contains three "empty" vacuoles, possibly extruded lamellar bodies. PN1at right is slightly more electron dense. Z15701: Multiple PNls lining alveolar spaced. No lamellar bodies or PN2s evident. Capillary at lower right with endothelial cells. Z15702: Cell type containing central lamellar body not clearly discernible. It appears to be outside the capillary and endothelial cell at topleft, but compressedbetween the pneumocyte on right and lower central cell. This is anarea of collapsed alveolar septa. Z15703: Alveolus and lining cells. Large PN1 appears to contain a single lamellar body in center of photograph. Capillary andendotheliumat left of photograph. Collapsed alveolar septain bottom center and right. Dilated cristae in some mitochondria. Z15704: 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.
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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: RCarht CDOBR varpiusc*)
Tissue:____ Liver___________
Block No(s).: 99.46-5________ Z 15773 -
Photo/Negative Nofsl.: Z15777
Significant Lesions (checkone): ______ Yes
X No
Interpreting Pathologist (signature anddate): ______________
Features: Z15773: 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 andsome dilation of the nuclear envelope. Six (6) peroxisomes counted. Z15774: Hepatocyte surroundedby some hematopoietic cells. Marked dilation and blebbing of RER. Four (4) peroxisomes counted. Z15775: Hepatocyte with canaliculus at the bottomand sinusoidal erythrocyte at top right. Endothelial cells poorly-defined andfixation is suboptimal. Eleven (11) peroxisomes counted.
Z15776: Hepatocyte is poorly-fixed and disrupted. Marked dilation andblebbing of the RER. Mitochondria are dense andcontain some granular deposits and dilated cristae. Seven (7) peroxisomes counted. Z15777: 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 of 5 cells)
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TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION
Study No.: 418-014: EM-99.46
Animal No.: 18912-1________ Treatment Group: 0 mg/kg. Control Sex: non applicable_______
Species/Strain: CRrait:Crxe>BR vaf/pius
Tissue: Lung______________
Block No(s).: 99.46-6_______ Z15768 -
Photo/Negative No(s).: Z15772
Significant Lesions (check one): ______ Yes
X No
Interpreting Pathologist (signature and date): ______________
Features: Z15768: 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. Z15769: Again, several cuboidal epithelial cells, consistent with bronchiolar epithelium. The two central cells contain a few lamellar bodies andare probably PN2s. The smaller darker cell appears to be a basal cell. Z15770: Alveolus with lining cells andunderlying collapsed alveolar septa. The small dark lining cell contains four lamellar bodies andis aPN2. The other two lining cells are PNls. Z15771: Area seems to be a terminal bronchiolejunctioning 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 theright and left are capillaries. The right one contains a lamellar body in its lumen. Z15772: Two large cuboidal epithelial cells or pneumocytes anda PN1 lining cell (bottom). Some dilated cristae in mitochondria noted.
Conclusions: Appears normal. A few randomPN2s with lamellar bodies noted.
0 0 1 5 (L
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TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION
Study No.: _418-014; EM-99.46
Animal No.: 18915-1________ Treatment Group: 0 me/ke. Control Sex: non applicable______
Species/Strain: Crl:CPBR VAF/Plus
-Rl -
Tissue:____ Liver___________
Block No(s).: 99.46-7_______ Z15763 -
Photo/Negative No(s).: Z15767
Significant Lesions (check one): ______ Yes
X _ No
Interpreting Pathologist (signature and date): ______________
Features: Z15763: Hepatocyte in photograph has alarge areaof cytoplasmfilled with abundant glycogen-like material. Plasma membranes and endothelial cells are poorlydefined due to poor fixation. Mitochondria aredense and contain some dense deposits and dilated cristae. There is dilation of the RER, showing asroundblebs on cross-section. A bile canaliculus is present in lower left. Fourteen (14) peroxisomes counted. Z15764: Hepatocyte, similar toZ15763. Five (5) peroxisomes counted. Z15765: Hepatocyte. Poor fixation clearly evidenced by loss of endothelial cells lining sinusoids. Eight (8) peroxisomes counted. Z15766: Six (6) peroxisomes counted. Much of the cytoplasmcontains fine dispersed granules, probably mostly glycogen, but also consistent with poor fixation. Z15767: 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).
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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:
Cri:CDBR Rat
va f/pius<b
Tissue: Lung_____________
Block No(s).: 99.46-8________ Z15758 -
Photo/Negative No(s).: 15762____
Significant Lesions (check one): ______ Yes
X No
Interpreting Pathologist (signature anddate): ______________
Features: Z15758: Collapsed alveolar septa. Air space at bottomis lined by a thin electron-dense cytoplasmic margin containing a few organelles anda large dilated bleb. Two capillaries in the collapsed areacontain some lamellar structures free within the lumina. Lamellar bodies within pneumocytes or free in the air spaces are not evident. Z15759: Alveolus and septa, mostly collapsed. PNls line air spaces. Capillary at top left appears tocontain a lamellar body within theendothelial cell cytoplasm. Smaller dark nucleus in the center appears to be a nucleus of aPN1. Z15760: Area of collapsed septa, with air spaceto left. The space is lined by thin margin of PN1. Subjacent to this space is acapillary containing a large lamellar body within the lumen. At top right is a cell, probably a PN2, whichcontains asingle lamellar body in its cytoplasm. The clear cell in thecenter appears to be apneumocyte. Z15761: Photograph contains several capillaries in the collapsed septa. The large lumen at the bottomcontains somepale-staining lipid material intermixed with endothelial cell cytoplasm. Air space is lined by flattened PN1 cytoplasm No lamellar bodies noted in pneumocytes. This profile in lumen may be tubular surfactant (lamellar body). Z15762: Central dark cell appears to be aPN2 with a few lamellar bodies in its cytoplasm. Flattened PN1 cytoplasmlines the remainderof the air space. Two capillary lumina, dorsal to and to the right of this PN2, contain some lamellar material. A focus of 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, anda single focus of lamellar material was present in airway lumen.___________________________
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TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION
Study No.: 418-014; EM-99.46
Animal No.: 18915-2________ Treatment Group: 0 me/ke. Control Sex: non applicable______
Species/Strain: RCralt:CD<S>BRVAF/Plus
Tissue: Liver____________
Block No(s).: 99.46-9__________ Z15753 -
Photo/Negative No(s~).: Z15757
Significant Lesions (check one): ______ Yes
X No
Interpreting Pathologist (signature and date): ______________
Features: Z15753: 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 andsome dilated cristae. Nine
(9) peroxisomes counted. Z15754: Hepatocyte surroundedby erythrocytes andhematopoietic cells. Endothelial cells are poorly defined due topoor fixation. Hepatocyte contains many dilated segments and blebs of RER and nuclear envelope. Five (5) peroxisomes counted. Z15755: Large central hepatocyte with abundant glycogen. Blebbing of RER abundant. Two (2) peroxisomes counted. Z15756: Field with several hepatocytes, however, only one at top center with nucleus is a complete cell. Cells has numerous dilated blebs of RER, some dark andsome light staining. Some dilated cristae notedin mitochondria. Four (4) peroxisomes counted. The cell at lower right appears tohave more peroxisomes, but is an incomplete cell for counting. Z15757: Several hepatocytes, althoughcentercell with nucleus is only complete cell. It has dense mitochondria with granular densities and numerous dilated blebs of RER. Twelve (12) peroxisomes counted.
Conclusions: Average number of peroxisomes = 6.4 (5 cells counted). Suboptimal fixation.
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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 v a F/PIus -Eat
Tissue: Lung ___________
Block No(s).: 99.46-10_______ Z 15748 -
Photo/Negative No(s).: Z15752
Significant Lesions (check one): ______ Yes
X____ No
Interpreting Pathologist (signature anddate): _______________
Features: Z15748: Two alveoli with intervening septumand capillary. PNls line the septum, andthere is alarge cell attached to theright. 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, andcollapsed septa. Lining pneumocyte contains a single lamellar body in its cytoplasmjust beneath the nucleus. At central top are several lamellar bodies with cytoplasmof probably PN2s. Z15750: Cuboidal epithelium of probably bronchiole. Centrally is aPN2 with 5 lamellar bodies.
Z15751: Alveolus and lining PMIs. At lower right is acapillary with a lamellar body in the lumen of the capillary. At top left, in collapsed septal area, is a PM2 with several lamellar bodies.
Z15752: Alveolus on left with lining pneumocytes is free of 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.
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TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION
Study No.: 418-014: EM-99.46
Animai No.: 18952-1_________ Treatment Group: 1.6 me/ke. high-dose Sex: non applicatile______
Species/Strain: Crl:CD<s>BRvaf/piu^ Rat
Tissue: Liver_____________
Block No(s).: 99.46-15_______ Z I5722 -
Photo/Negative No(sV. Z I5726
Significant Lesions (checkone): X Yes ______ No
Interpreting Pathologist (signature and date): ______________
Features: Z15722: Hepatocyte. Cytoplasmcontains abundant fine granular material, glycogen. Some dilated blebs of RER. Mitochondria appear dense with granular deposits andsome dilated cristae. Twenty (20) peroxisomes identified. Adjacent endothelial cell is poorly-defined (poor fixation). Z15723: Similar to Z15722. Numerous round dilated blebs of RER. Bile canaliculus present in bottomof photograph. Ten (10) peroxisomes counted. Z 15724: Sinusoidcontaining several erythrocytes. Segment of hematopoietic granulocytic cell in top left. Endothelial cell and lipid-containing perisinusoidal cell are poorly fixed. Two incomplete hepatocytes onright. A few peroxisomes are noted, but not counted. Z15725: Hepatocyte. Bile canaliculus at top; sinusoid in lower right of photograph. Numerous dilated blebs of RER. Fifteen (15) peroxisomes counted. Z15726: 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.
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TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION
Study No.: 418-014: EM-99.46
Animai No.: 18952-1________ Treatment Group: 1.6 mg/kg. high-dose Sex: non applicable______
Species/Strain: Cri:CD<a?BR v a f /p1us(B> JUi
Tissue: Lune ___________
Block No(s).: 99.46-16______ Z15717-
Photo/Negative No(s).: Z15721
Significant Lesions (check one): X Yes ______ No
Interpreting Pathologist (signature anddate): ______________
Features: 15717: Alveolus with lining cells. Two large PNIs arepresent and appear to have a PN2 between them. However, aPN1 margin still separates the PN2 from the sinusoid. Several lamellar bodies are present associatedwith the PN2, and some of them appear to be extracellular. The PN1 on the left contains abundant fine granular material in its cytoplasm, consistent with glycogen. Z15718: Collapsed area of alveolar septa. Two central cells contain lamellar bodies, three (3) in one, and seven (7) in the other. Both cells appearto be PN2s, although normal pulmonary architecture is unclear because of the collapsed state. Z15719: 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 aPN2. A large lamellar body is associated with the left cell, but it may beextracellular. Both cells contain abundant granular material, consistent with glycogen. Z15720: Alveolus and lining cells. Two cells contain lamellar bodies, six in one and eight in the other. Some lamellar material appears tobe extracellular. Z15721: Alveolus, five lining cells, and acapillary in top right of photograph. This is possibly ajunction with a terminal bronchiole. Two of thepneumocytes contain multiple lamellar bodies, and appear to be PN2s.
Conclusions: There appear tobe increased numbers ofPN2s containing lamellar bodies. Some extracellular lamellar material noted.
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418-014:PAGE F-22
TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION
Study No.: 418-014: EM-99.46
Animal No.: 18952-2__________ Treatment Group: 1.6 mg/kg. high-dose Sex: non applicable______
Species/Strain: Crl:CD(B)BRVAF/Plus
Rat
Tissue: Liver____________
Block No(s).: 99.46-17______ Z15712-
Photo/Negative No(s).: Z15716
Significant Lesions (checkone): X Yes ______ No
Interpreting Pathologist (signature anddate): ______________
Features: Z15712: Hepatocyte. Poorly-defined sinusoids and endothelium at top right andbottomof photograph. The hepatocyte cytoplasmappears voluminous with abundant fine granular material consistent with glycogen. Mitochondria appear dense and have some granular deposits and dilated cristae. Some dilated blebs of RER. Eight (8) peroxisomes identified. Z15713: Hepatocyte. Hematopoietic cell to right. Bile canaliculus present in lower right. Abundant fine granular material, glycogen, in cytoplasm. Appear to be increased numbers of peroxisomes. Twenty-seven (27) peroxisomes present. Z15714: Hepatocyte. Many dilated blebs of RER and abundant glycogen particles in cytoplasm. Fourteen (14) peroxisomes identified in this cell. Z 15715: Hepatocyte. Many dilated blebs of RER and abundant glycogenparticles in cytoplasm. Mitochondria with some granular deposits anddilated cristae. Nineteen (19) peroxisomes identified in this cell. Z15716: Sinusoid with lining endothelial cell. Cellular fixation is poor. Adjacent hepatocytes contain many mitochondria, RER and glycogen-like particles. Dilated blebs of RER are present. Multiple peroxisomes are present, but not counted because whole hepatocytes are not in photograph.
Conclusions: Hepatocytes appear to contain increased numbers of peroxisomes, average 17/hepatocyte (4 cells counted). Cells contain abundant glycogen. Suboptimal cellular fixation.
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TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION
Study No.: 418-014: EM-99.46
Animai No.: 18952-2________ Treatment Group: 1.6 mg/ke. high-dose Sex: non applicable_______
Species/Strain: Crl:CD(5>BRVAF/Plus
Rat
Tissue: Lune______________
Block No(s).: 99.46-18______ Z15707 -
Photo/Negative No(s).: Z15711
Significant Lesions (checkone): X
Yes ______ No
Interpreting Pathologist (signature anddate): ______________
Features: 15707: SinglePN2 lining alveolus. Cell containsmultiple mitochondria aroundthe nucleus, andalsomultipleclear vacuoles, whicharemostlyempty ofcontents. Thesewere probably lamellarbodieswhichwereextrudedintothealveolar space. These vacuoles are lined by asingle membrane, whereasthe mitochondriahaveadoublemembrane. 15708: Collapsedalveolar septa. Athin layeredPN1 coversthesurface. Beneathisan endothelial cell with manypinocytotic vesicles, oneasinglelamellarbody. Centrally isanarea of flattened basement membrane material containingmanycollagen fibrils. A darkcell ispresent in theupper right, whichcontainsmanymitochondria. To theleft of this cell aretwo lamellar bodies, althoughthey appear tobeinacytoplasmicextensionof anunidentifiedcell. Z15709: A nucleatedPN2 containingmultiple lamellarbodies(5) appearsto beoverlain by a PN1 lining thealveolus. Z15710: Alveolar spacein topright islinedbyathinPN1cell. Adjacent tothis PN1and coveredby it'scytoplasmisanextracellularareawithseveral (3) lamellar bodies. A capillary is dorsal containingan erythrocyteandendothelial cell. Several cells within collapsed septaarenot clearly identifiable astocell type. Z15711: A singledarkPN2 issqueezedbetweentwoPNls. Itscytoplasmcontains several empty vacuoles, which have apparently extrudedlamellarbodies. To the right ofthis cell are two large lamellar bodieswithin somelipid-like material. Thesebodiesmaybe in theadjacent PN1, macrophage, or maybeextracellular. A small capillary isinthelowerright margin of the
photograph.______________________________________________________
Conclusions: There appear to be increased numbers of type II pneumocytes (PN2s), and more lamellar bodies were seen in this sample. Otherwise, cellular morphology appeared essentially normal. Fixation appeared satisfactory.
ooisl
418-014:PAGE F-24
TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION
Study No.: 418-014: EM-99.46
Animai No.: 18956-1_____________ Treatment Group: 1.6 mg/ke. high-dose Sex: non applicable_______
Species/Strain: Crl:CDtRRVAF/PiusOt -Rat
Tissue: Liver____________
Block No(s).: 99.46-13______ Z15732-
Photo/Negadve No(s).: Z15736
Significant Lesions (check one): X
Yes ______ No
Interpreting Pathologist (signature and date): _______ ;______
Features: Z15732: Hepatic sinusoidcontaining nucleatederythroycte and acouple mature erythrocytes. Some lamellar debris is present in the sinusoid. Two endothelial cell nuclei are present. There are nocomplete hepatocytes in this photograph. Z15733: 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. Z15734: Hepatocyte. Similar toZ15733. Numerous dilated blebs of RER are present Twenty-four (24) peroxisomes. ZI5735: Hepatocyte with similar morphology asdescribed for above cells. Sixteen (16) peroxisomes counted. Z15736: Hepatocyte. Dilated blebs ofRER present Mitochondria are dense with granular deposits and dilated cristae. Eighteen (18) peroxisomes.
Conclusions: Increased numbers of peroxisomes per hepatocyte. 17.25 peroxisomes per hepatocyte (average of four counted cells). Suboptimal fixation.
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TRANSMISSION ELECTRON MICROGRAPH INTERPRETATION
Study No.: 418-014; EM-99.46
Animai No.: 18956-1________ Treatment Group: 1.6 me/kg. high-dose Sex: non applicable_______
Species/Strain: CrhCTXBBR VAF/Plus
Rat
Tissue:_____ Lune _________
Block No(s).: 99.46-14______ Z I5727 -
Photo/Negative No(s).: Z15731
Significant Lesions (check one): X Yes ______ No
Interpreting Pathologist (signature anddate): ______________
Features: 15727: Alveolus with adjacent collapsed septa. The two bottompneumocytes are PN2 with lamellar bodies. Someof the bodies arepartially 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 acapillary with a nucleated erythrocyte. Z15728: Collapsed alveolar septa. Several cells contain lamellar bodies. At left one lamellar body appears to be within the lumen of acapillary. Z15729: Somewhat T-shaped capillary containing anelectron-dense erythrocyte. The endothelial cell nucleus is ventral to the erythrocyte. Within the lumen of the capillary are several lamellar bodies. At left of field is the thin cytoplasmic lining of a PN1. The other cells appear to be sections through pneumocytes at various angles. Z15730: Alveolar lumen and lining cells. Centrally is asmall PN2 containing a couple small lamellar bodies next to its nucleus. The dark-staining cytoplasm to the right of this cell appears to be an extension from analveolar macrophage. To the left are acouple pneumocytes, andthey contain acouple lamellar bodies. Z15731: 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 of pneumocytes containing lamellar bodies. Some lamellar bodies noted in capillaries.
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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: Crl:CDBR VAF/Plus
Rat
Tissue: Liver____________
Block No(s).: 99.46-11______ Z15742 -
Photo/Negative No(s').: Z 15747
Significant Lesions (check one): X Yes ______ No
Interpreting Pathologist (signature anddate): ______________
Features: Z 15742: Hepatocyte contains abundant fine granular material consistent with glycogen in the cytoplasm. There are numerous dilated blebs of RER on cross section. Mitochondria appear dense andhave some granular deposits. Seventeen (17) peroxisomes counted. Some lamellar material notedextracellularly andpossibly within clear spaces in the left portion of the hepatocyte. Z 15743: Hepatocyte, overall similar to Z15742, although fixation appears better. Nine (9) peroxisomes counted. Lamellar material at top appears to be extracellular. Z15744: Hepatocyte. At left is ahematopoietic cell in sinusoid and at upper right appears to be a dilated bile canaliculus. Abundant area of glycogen in cytoplasm. Nineteen (19) peroxisomes counted. Z 15745: Sinusoid, endothelial cell and incomplete portions of acouple hepatocytes. Centrally is aclear cytoplasmic areawith many dilated blebs of 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 of RER on cross section. Mitochondria appear dense and have some granular deposits and some dilated cristae. Eleven (11) peroxisomes counted. Z15747: 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.
0015
418-014:PAGE F-27
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: CrkClxaBR VAF/Plus
Rat
Tissue: Lung_____________
Block No(s).: 99.46-12______ Z15737 -
Photo/Negative NoCsl.: Z15741
Significant Lesions (check one): X Yes ______ No
Interpreting Pathologist (signature anddate): ______________
Features: Z15737: 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 of acell'scytoplasmthat appears to contain some lamellar body vacuoles. A couple PNls are present adjacent to the PN2, and acapillary with erythrocyte andendothelial cell ispresent in lower left central area of photograph. Z15738: Alveolus with lining cells. The large nucleus belongs to a PN1 whose cytoplasmlines the air space. Beneaththis cell andjust to theright are several lamellar bodies, although it'sin asubjacent capillary. To the middle left is acapillary with an eyrthrocyte that does contain some lamellar debris in the lumen. Z15739: Alveolus and collapsed septa. Similar to Z15738, some lamellar bodies appear to be within the lumen of a capillary. PNls line the alveolar space. Z15740: Large capillary with several erythrocytes to the right. Left of this is acellular extension, probably aPN2, that contains a distinct lamellar body. A thin layer of PN1 lines the alveolar space. Z15741: Alveolar space contains abundant lamellar material. Several PNls 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 of the photograph.____________
Conclusions: Appear to be increased lamellar bodies in the tissuecompared with control sample, although many were in capillary lumina andsome in alveolar and interstitial spaces.
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418-014:PAGE F-28 DI. Transmission Electron Micrographs
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418-014:PAGE F-29 NOTE: The original glossy figure photographs (Figures 1-12) were delivered to Dr. Marvin T. Case with the draft pathology report on March 19, 1999.
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PrimedicaAAncriglulasryStPuadtyhoNloog. y41R8e-0p1o4n
Figure Legends
Figure 1. Liver. Hepatocyte showing normal cytoplasmic organelles. White spots within mitochondria represent some dilatation of cristae. Some peroxisomes are identified. To the right is anendothelial cell anderythrocytes of adjacent sinusoid. Animal No. 189041, 0 mg/kg/day (control), 11.200X magnification.
Figure 2. Liver. Hepatocyte. Dilatation of nuclear envelope andrough 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 sinusoidat upper right of photograph. Animal No. 18912-1, 0 mg/kg/day (control), 11,200X magnification.
Figure 3. Liver. Cellular morphology is similar toFigure 2. Some peroxisomes are labeled, and appear more numerous than in controls. A bile canaliculus is present in upper right of photograph and hematopoietic cells arepresent in sinusoid at upper left. Animal No. 18952-2, 1.6 mg/kg/day (high-dose), 11,200X magnification.
Figure 4. Liver. Dilatation of rough endoplasmic reticulumis evident. Some peroxisomes, increased in number, arelabeled. Animal No.18956-1, 1.6 mg/kg/day (high-dose), 11,200X magnification.
Figure 5. Liver. Hepatocyte demonstrating somewhat better fixation with more even dispersion of organelles. Clear space at top of 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 cytoplasmidentify this cells asa 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 of the airway space. Two small lamellar bodies in one cuboidal cell suggest this is atype II pneumocyte which has secreted most of its lamellar bodies and is destined tobecome a type I pneumocyte. Animal No. 189042, 0 mg/kg/day (control), 11.200X 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 of the septal wall both above andto the right of the type II pneumocyte. Animal No. 18915-1,0 mg/kg/day (control), 11.200X magnification.
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418-014:PAGE F-31
PrimedicaAAncriglluasryStPuadtyhoNloog. y41R8e-p0o1n4
Figure 9. Lung. Dense-staining ceil is a type II pneumocyte positioned between two type I pneumocytes. Four distinct lamellar bodies arepresent in the cytoplasmof the type 13 pneumocyte, although the lamellar material is partially washed out in these vacuoles. To the right of this cell are two dark lamellar-type bodies surrounded by some lipid. This appears to be within acell in acollapsed area of septa, andmay be amacrophage. Animal No. 18952-2, 1.6 mg/kg/day (high-dose), 11,200X magnification. Figure 10. Lung. Collapsed areaof septashowing 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 of thephotograph. Animal No. 189521, 1.6 mg/kg/day (high-dose), 11.200X magnification. Figure 11. Lung. Centrally is acell containing several lamellar-like structures which appears tobe atype II pneumocyte. Glycogen-like material appears to fill much of the cytoplasm. Adjacent cells to the right containcentrioles andrepresent 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 secretedfrom type II pneumocytes. A type I pneumocyte lines the space on the left andright. Two cuboidal cells containing glycogen-like granular material appear to be pneumocytes, probably type 13swhich have released their lamellar bodies. Animal No. 18973-1, 1.6 mg/kg/day (high-dose), 11.200X magnification.
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418-014:PAGE F-32
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
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 El pneumocyte Plasma membrane Rough endoplasmic reticulum
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c e is 3 i
418-014:PAGE F-45 Quality Assurance Statement
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Pathology Associates International
A Company of Science Applications International Corporation
QUALITY ASSURANCE STATEMENT
This electron microscopy study has been inspected and audited by the Quality Assurance Unit as required by the Good Laboratory Practices Regulations promulgated by the U.S. Food and Drug Administration. PAI has a functioning and responsive Quality Assurance Unit which reports directly to management. The following is a record of inspections/audits and their resulting reports:
Date.oi
Inspection
Phase Inspected
Date Findings Reported
to Management and-Study Director
2/18,19/99 3/16/99 3/17/99 4/5/99
Processing and Embedding Data Audit Draft Report Audit Final Report Audit
2/19/99 3/17/99 3/17/99 4/5/99
Sponsor: 3M Corporate Toxicology Test Article: PFOS Study Number: 418-014, PAI Study EM-99.46
001532
4915 D Prospectus Drive Durham , North Carolina 27713 (919) 5 4 4 -5 2 5 7 * (91 9) 5 4 4 -3 2 1 8 FAX
APPENDIX G STATEMENT OF THE STUDY DIRECTOR
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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 practice0occurred that affected the quality or integrity of the study.
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.
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APPENDIX H QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT
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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.
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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.
Quality Assurance Manager
Quality Assurance Supervisor and Principal Auditor
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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
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 Pups Dates of inspection: 28 JAN 99, 28 JAN 99 Dates results reported to the Study Director and Management: 02 FEB 99, 02 FEB 99
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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.
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