STUDY NO. 00-4204 

SPONSOR STUDY NO. 211-ETBE-1G 

GASOLINE ETBE VAPOR CONDENSATE: 

A ONE-GENERATION WHOLE-BODY INHALATION REPRODUCTION 

TOXICITY STUDY IN RATS 

Final Report

Volume II of II 

Submitted to: American Petroleum Institute 

 1220 L Street, Northwest 

 Washington, D.C. 20005-4070 

 Attn: Thomas M. Gray, M.S., D.A.B.T. 

 Date: 18 July 2011 

Page 513 of 845 
Huntingdon Life Sciences 00-4204 Page 514 

 211-ETBE-1G Final Report

TABLE OF CONTENTS 

Volume II 

COVER PAGE .................................................................................................................513

TABLE OF CONTENTS .................................................................................................514 

APPENDICES 

Z. Individual Animal Gross and Microscopic Observations .......................................... 515 

AA. Released Pup and Historical Control Data ................................................................. 719 

BB. Sperm Analysis Report ............................................................................................... 752 

CC. Certificates of Analysis – (Feed, Bedding, and Water) .............................................. 767 

DD. Protocol and Protocol Amendments ........................................................................... 808 

EE. Testing Facility Personnel ........................................................................................... 844 

FF. Report Amendments .................................................................................................... 845 
Huntingdon Life Sciences 00-4204 Page 515 

 211-ETBE-1G Final Report

 Individual Animal Gross 

and Microscopic Observations 

Preface Appendix Z 

Key to Abbreviations 

Gl = Gland 

LN = Lymph Node 

Oviducts/Fallop = Oviducts/Fallopian 

Notes

1. Unless otherwise specified in a histopathology note, the organ/tissue 

examined was the required (routine) section. 

2. The dimensions of gross lesions are presented as a general range based on 

the actual dimensions noted during macroscopic examinations. The actual 

dimensions of gross lesions can be found in the raw data. 

Corresponding exposure levels for each group were as follows: 

Group 1 - 0 mg/m3 

Group 2 - 2000 mg/m3

Group 3 - 10000 mg/m3

Group 4 - 20000 mg/m3











































































































































































































Huntingdon Life Sciences 00-4204 Page 719 

 211-ETBE-1G Final Report

 

Released Pup and Historical Control Data Appendix AA

Note 

During Lactation Days 25-27 on 18 July 2002, a Group 1 rack containing the pups 

from dam numbers 1554, 1556, 1560, 1563, 1565, 1566, 1568, 1569, 1572, 1574, 

1575, and 1576 was accidentally overturned releasing the litters so that it could not 

be determined which pups were from which litters. The dams were in the exposure 

chamber at the time of the incident and were unaffected. Of the 12 litters of 

control-group pups, 108 pups in total were released and 105 survived. The 

laboratory necropsied and collected organ weight data on all 105 pups. 

In addition, one female pup missing from dam number 1552 on Lactation Day 28 

was apparently mis-identified as a released pup. Organ weights were collected and 

the pup was necropsied with the released pups. 

The data for all 106 pups was separated according to sex, and the mean, 

standard deviation and range of values is presented separately from data of 

unaffected litters. The corresponding Lactation Day (LD) for each litter on the 

day of necropsy and data collection is as follows: 

1554 LD27 1556 LD28 1560 LD28 1563 LD26 

1565 LD26 1566 LD27 1568 LD27 1569 LD27 

1572 LD27 1574 LD27 1575 LD27 1576 LD27 

Table of Contents 

Released Pup Data 

 Mean Body Weight Values ............................................................................. 720 

 Summary of Pup Necropsy Observations ....................................................... 722 

 Summary of Absolute Organ Weights ............................................................ 725 

 Summary of Organ Weight to Body Weight Ratio ........................................... 727 

 Summary of Organ Weight to Brain Weight Ratio .......................................... 729 

 Individual Body Weight Values ....................................................................... 731 

 Individual Pup Necropsy Observations ........................................................... 735 

 Individual Organ and Final Body Weights ....................................................... 743 

Historical Control Data 

 Summary of Mean Pup Body Weights/Litter ................................................... 747 

 Summary of Pup Necropsy Observations ....................................................... 748 

 Summary of Mean Pup Organ Weights: 

 Male Organ/Body and Organ/Brain Weight Ratios (%) ................................ 750 

 Female Organ/Body and Organ/Brain Weight Ratios (%) ............................ 751 



























Huntingdon Life Sciences 00-4204 Page 747 

 211-ETBE-1G Final Report 

 Historical Control Data 

Summary of Mean Pup Body Weights/Litter Appendix AA 

Study No. Lactation Day 

 1 4 7 14 21 28

 

 

00-4202 Mean 7.2 10.4 14.5 26.0 42.5 78.8

 S.D. 0.52 1.03 1.23 1.68 3.87 5.12

 N 23 22 22 22 22 22

 

00-4203 Mean 7.3 10.6 14.7 25.6 41.3 76.7

 S.D. 0.48 0.98 1.50 2.25 3.97 6.16

 N 25 25 25 25 25 25

 

00-4207 Mean 7.0 10.4 14.6 24.4 40.7 69.5

 S.D. 0.75 1.36 1.70 2.31 3.99 7.40

 N 24 24 24 24 24 24

 

00-4208 Mean 6.9 9.7 13.5 23.7 38.7 72.6

 S.D. 0.67 0.98 1.13 1.78 3.47 5.60

 N 23 23 23 23 23 23

 

Overall Mean 7.1 10.3 14.3 25.0 40.8 74.3

 S.D. 0.64 1.13 1.48 2.20 4.01 7.04

 N 95 94 94 94 94 94
Huntingdon Life Sciences 00-4204 Page 748 

 211-ETBE-1G Final Report 

 Historical Control Data 

Summary of Pup Necropsy Observations Appendix AA 

Study Number 00-4202 00-4203 00-4207 00-4208 Total

Rat Strain: 

Sprague-Dawley [Crl: CD IGS BR] 

 

 

Total # male pups 114 117 117 137 485 

Total # female pups 127 112 117 100 456 

Total # undetermined 0 1 0 0 1 

 

Stillborn 2 5 8 1 16 

Lungs: floatation test - stillborn negative 2 5 6 1 14 

Stomach: no milk present 0 4 6 1 11 

Stomach: milk undetermined 2 1 2 0 5 

Gross exam: autolysis 2 0 0 0 2 

Gross exam: cannibalized 1 1 2 0 4 

Gross exam: internal sex undetermined 1 1 0 0 2 

Abdominal cavity: fluid filled 0 1 0 0 1 

Skin: discolored area 0 1 0 0 1 

Skin: edematous - general 0 1 0 0 1 

 

Died prior to Cull 16 0 7 6 29 

No remarkable observation 14 0 0 0 14 

Lungs: floatation test - found dead, 

positive 

0 0 2 2 4 

Stomach: milk present 1 0 3 4 8 

Stomach: no milk present 0 0 3 1 4 

Head: skull damaged 0 0 0 0 0 

Gross exam: autolysis 1 0 1 1 3 

 

Day 4 Cull 93 75 80 97 345 

No remarkable observation 93 75 80 97 345 

 

Died prior to 28 Day Sacrifice 0 0 1 0 1 

Head: skull damage 0 0 1 0 1 

Stomach: milk present 0 0 1 0 1 

 

Day 28 Sacrifice 130 150 138 133 551 

No remarkable observation 127 136 130 126 519 

Epididymides: small 0 0 0 1 1 

External exam: left hindpaw missing 0 1 0 0 1 

Eyes: damaged 0 0 1 0 1 
Huntingdon Life Sciences 00-4204 Page 749 

 211-ETBE-1G Final Report 

 Historical Control Data 

Summary of Pup Necropsy Observations Appendix AA 

Study Number 00-4202 00-4203 00-4207 00-4208 Total

Rat Strain: 

Sprague-Dawley [Crl: CD IGS BR] 

 

 

Gross exam: cannibalized 0 0 2 0 2 

Kidney: cystic 3 6 3 6 18 

Kidney: dilated pelvis 0 6 0 1 7 

Liver: discolored foci 0 0 1 0 1 

Skin: alopecia - general 0 0 1 0 1 

Testes: small 0 0 0 1 1 

Testes: undescended, unilateral 0 1 0 0 1 
Huntingdon Life Sciences 00-4204 

 211-ETBE-1G 

Males 

Historical Control Data 

Summary of Mean Pup Organ Weights 

 TBW=Terminal Body Weight; ORG=Organ Weight 

Study TBW Brain Spleen Thymus 

No. Wt. ORG/ 

TBW 

WT ORG/ 

TBW 

ORG/ 

BRAIN 

WT ORG/ 

TBW 

 (g) (g) (%) (g) (%) (%) (g) (%) 

00-4202 Mean 82.3 1.5825 1.9312 0.3245 0.3951 20.514 0.3586 0.4392 

 S.D. 6.4 0.0629 0.1263 0.0500 0.0588 3.142 0.0547 0.0512  N 22 22 22 22 22 22 22 22 

00-4203 Mean 81.8 1.6118 1.9785 0.3445 0.4210 21.381 0.3673 0.4499 

 S.D. 6.8 0.0713 0.1305 0.0477 0.0472 2.845 0.0887 0.1105  N 25 25 25 25 25 25 25 25 

00-4207 Mean 72.7 1.5939 2.2127 0.2792 0.3850 17.589 0.3238 0.4471 

 S.D. 8.1 0.1572 0.2845 0.0491 0.0613 3.031 0.0788 0.1076  N 23 23 23 23 23 23 23 23 

00-4208 Mean 75.6 1.5634 2.0757 0.3218 0.4258 20.582 0.3322 0.4389 

 S.D. 5.6 0.0591 0.1267 0.0484 0.0589 3.031 0.0600 0.0718  N 22 22 22 22 22 22 22 22 

Overall Mean 78.2 1.5888 2.0490 0.3180 0.4067 20.035 0.3459 0.4440 

 S.D. 7.9 0.0967 0.2077 0.0537 0.0584 3.300 0.0737 0.0885  N 92 92 92 92 92 92 92 92 
Huntingdon Life Sciences 00-4204 

 211-ETBE-1G 

Females 

Historical Control Data 

Summary of Mean Pup Organ Weights 

Study TBW Brain Spleen Thymus 

No. Wt. ORG/ 

TBW 

Wt. ORG/ 

TBW 

ORG/ 

BRAIN 

WT ORG/ 

TBW 

B

 (g) (g) (%) (g) (%) (%) (g) (%) 

00-4202 Mean 76.7 1.5323 2.0070 0.2884 0.3741 18.875 0.3836 0.5005 

 S.D. 5.5 0.0671 0.1472 0.0681 0.0702 4.548 0.0468 0.0486  N 22 22 22 22 22 22 22 22 

00-4203 Mean 75.1 1.5506 2.0750 0.3043 0.4041 19.597 0.3459 0.4384 

 S.D. 6.36 0.0631 0.1576 0.0559 0.0606 3.307 0.0466 0.1045  N 25 25 25 25 25 25 25 25 

00-4207 Mean 67.4 1.5015 2.2454 0.2440 0.3613 16.388 0.3253 0.4869 

 S.D. 6.87 0.1191 0.2698 0.0473 0.0564 3.713 0.0884 0.1423  N 23 23 23 23 23 23 23 23 

00-4208 Mean 69.9 1.5284 2.1971 0.2831 0.4058 18.506 0.3420 0.4891 

 S.D. 5.90 0.0688 0.1724 0.0428 0.0573 2.541 0.0607 0.0805  N 22 22 22 22 22 22 22 22 

Overall Mean 72.3 1.5287 2.1305 0.2803 0.3867 18.3614 0.3488 0.4775 

 S.D. 7.1 0.0833 0.2120 0.0580 0.0633 3.7352 0.0653 0.1020  N 92 92 92 92 92 92 92 92 
Huntingdon Life Sciences 00-4204 Page 752 

 211-ETBE-1G Final Report

 

Sperm Analysis Report Appendix BB
AUDITED FINAL REPORT 

Study Phase: Sperm Analysis

Test Site Phase Reference Number 93075219 

Testing Facility Study Number 00-4204 

Sponsor Reference Number 211-ETBE-1G 

Gasoline ETBE Vapor Condensate: 

A One-Generation Whole-Body Inhalation Reproduction 

Toxicity Study in Rats 

TEST SITE: 

Charles River Laboratories, Pathology Associates - Maryland 

15 Worman’s Mill Court, Suite I 

Frederick, MD 21701 

TESTING FACILITY: 

Huntington Life Sciences (HLS) 

Mettlers Road 

P.O. Box 2360 

East Millstone, NJ 08875-2360 

SPONSOR: 

American Petroleum Institute (API) 

1220 L Street, Northwest 

Washington, D.C. 20005-4070 

09-APR-2009 

Page 1 of 14 

Page 753
Final Report 

HLS Study No.: 00-4204 

2

 TABLE OF CONTENTS 

Section Page No.

I. QUALITY ASSURANCE STATEMENT .............................. 4

II. MATERIALS AND METHODS .................................... 5

A. Sample Collection................................... 5

B. Sperm Motility Evaluation........................... 5

C. Caudal Epididymis Weight and Total Sperm Count 

Determination ...................................... 5

D. Sperm Morphology Evaluation......................... 6

E. Statistical Analyses................................ 6

F. GLP Compliance Statement............................ 6

G. Sample and Data Archiving........................... 7

III. RESULTS .................................................. 7

A. Sperm Motility...................................... 7

B. Total Sperm Count................................... 7

C. Sperm Morphology.................................... 7

D. Caudal Epididymis Weight............................ 7

IV. DISCUSSION AND CONCLUSIONS ............................... 8

V. REFERENCES ............................................... 9

Page 754
Final Report 

HLS Study No.: 00-4204 

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INDEX OF TABLES AND APPENDICES 

Table Page No.

 1. Summary of Male Reproductive Parameters .................10 

Appendix Page No.

A. Individual Caudal Epididymis Weight, 

 Sperm Motility and Sperm Count Data .....................11 

B. Individual Sperm Morphology Data .........................13 

Page 755

Final Report 

HLS Study No.: 00-4204 

5

Gasoline ETBE Vapor Condensate: 

A One-Generation Whole-Body Inhalation Reproduction Toxicity 

Study in Rats 

 II. MATERIALS AND METHODS 

 A. Sample Collection

 All surviving male rats were euthanized by exsanguination 

following anesthesia with inhaled carbon dioxide after the last F1

litters were delivered. For all animals, the abdominal cavity was 

opened and the reproductive organs exposed. For motility 

assessment, the left vas deferens was dissected away from the 

testis and immediately placed in a petri dish containing 10 ml of 

a solution consisting of 1% Bovine Serum Albumin dissolved in 

Phosphate Buffered Saline. The solution was prewarmed to a 

temperature of approximately 38o

C. A minimum 3-minute period was 

allowed for the sperm to swim out. 

 For total sperm count assessment, the left epididymis and 

left testis were then removed and placed on dry ice. The frozen 

epididymides and testes were then transferred to Pathology 

Associates - A Charles River Company (formerly known as Pathology 

Associates International until January 8, 2001), Frederick, MD. 

and stored frozen at -70oC until evaluation for caudal epididymal 

sperm count and homogenization-resistant testicular sperm count. 

 Only the control and high-exposure animals were examined for 

percent motility, total epididymal and testicular sperm count, 

sperm morphology and caudal epididymis weight. 

 B. Sperm Motility Evaluation

 Following the swim out period, a sperm sample was obtained 

using a 100 μm deep cannula. The cannula was immediately loaded 

into the prewarmed stage of the Hamilton Thorne IVOS automated 

sperm analyzer (version 12.1.c). The analyzer automatically 

selected five fields and each motion image was digitally saved 

and permanently stored on optical media. The images were 

subsequently analyzed and the percent motility determined for 

each animal. 

 C. Caudal Epididymis Weight and Total Sperm Count Determination

 Each frozen epididymis was removed from the freezer, thawed 

and the caudal section was trimmed and weighed. Each frozen 

Page 757
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HLS Study No.: 00-4204 

6

testis was removed from the freezer, thawed, the tunica removed 

and the testicular parenchyma weighed. The cauda epididymis and 

testicular parenchyma were homogenized in deionized water and the 

suspensions were transferred to plastic test tubes and vortexed. A 

100 μl sample was transferred to a violet reaction vial containing 

a Hoechst dye (H33342) which uniquely stains the head of the 

sperm. A sample of the stained sperm was placed into a 20 μm deep 

Cell-Vu glass slide which was loaded into the analyzer. Twenty 

fields were automatically selected by the analyzer for each animal 

and the number of homogenization-resistant sperm and cauda 

epididymal sperm determined. The counts reported were adjusted 

for caudal epididymal or testicular weight. 

 D. Sperm Morphology Evaluation

 Two Eosin stained slides were prepared for each animal from 

the caudal epididymis total count preparation. The slides were 

evaluated and a minimum of 200 sperm cells/animal was examined for 

morphological development. 

 E. Statistical Analyses

 The means and standard deviations for the sperm motility, 

caudal epididymal sperm count, testicular homogenization-resistant 

spermatid count and sperm morphology data were calculated and 

compared across groups using the Kruskal-Wallis nonparametric 

ANOVA test. If a significant effect occurred (p<0.05), the MannWhitney U test was used for pair-wise comparisons of each treated 

group to the control group. Animals with fewer than 25 sperm 

cells present in the motility analysis were excluded from the 

group mean calculation and subsequent statistical analysis. 

Animals with at least 25 sperm cells were included in the group 

mean calculation subsequent and statistical analysis; however, 

animals with fewer than 50 cells were identified to have a low 

count. The mean caudal epididymis weight for each group was 

calculated and compared using one-way analysis of variance (ANOVA) 

technique. If ANOVA is significant, Dunnett's test was used for 

pair-wise comparisons of each treated group to the control group 

at the 5% and 1% risk levels. Statistical analyses were 

performed using an IBMTM compatible computer with SAS computer 

programs (SAS/STAT User's Guide, 1989). 

 F. GLP Compliance Statement

 The portion of this study performed by PAI was conducted in 

compliance with US EPA 79.60, CFR Vol. 59, No. 122, 27 June 1994. 

Page 758
Final Report 

HLS Study No.: 00-4204 

7

 G. Sample and Data Archiving

 Frozen epididymis and testis samples not processed for sperm 

count determination were transferred to and stored frozen at -70o

C 

in PAI's long-term archive facility. The optical media used for 

permanent storage of the sperm motility images, raw data printouts 

from the sperm motility, epididymal and testicular sperm count and 

sperm morphology analysis, as well as any supporting documentation 

and the sperm morphology slides will be maintained at PAI until 

the study has been completed. Upon finalizing the report, all 

non-frozen study material will be returned to HLS for archiving. 

Disposition of the frozen tissue samples will be directed by 

correspondence from HLS. 

III. RESULTS 

 A. Sperm Motility

 Table 1 (Summary Data) 

 Appendix A (Individual Data) 

 No apparent treatment-related effects were observed in the 

sperm motility data. Group mean values were 96% and 94% for the 

control and high-exposure groups, respectively. 

 B. Total Sperm Count

 Table 1 (Summary Data) 

 Appendix A (Individual Data) 

 The number of sperm per gram of caudal epididymis and testis 

was comparable between the control and high-exposure groups. Group 

mean values were 956.7 and 1053.0 million sperm/gram for the 

epididymal sperm count and 105.7 and 106.5 million sperm/gram for 

the homogenization-resistant testicular count. 

 C. Sperm Morphology

 Table 1 (Summary Data) 

 Appendix B (Individual Data) 

 A low incidence of head/tail abnormalities was observed for 

animals in the control and high-exposure groups. Group mean 

values were 0.8 and 0.6 percent abnormal sperm. No treatment 

related differences were observed. 

 D. Caudal Epididymis Weight

 Table 1 (Summary Data) 

Page 759

Final Report 

HLS Study No.: 00-4204 

9

 V. REFERENCES 

Dunlap, W.P., Marx, M.S. and Agamy, G.G. 1981. Fortran IV 

functions for calculating probabilities associated with 

Dunnett’s test. Behav. Res. Methods and Instrumentation 13: 

363-366. 

Dunnett, C. W. 1955. A multiple comparison procedure for 

comparing several treatments with a control. Journal of the 

American Statistical Association 50:1096-1121. 

Dunnett, C. W. 1964. New tables for multiple comparisons with a 

control. Biometrics 20:482-491. 

Kruskal, W.H. and Wallis, W.A. 1952. Use of ranks in onecriterion variance analysis. Journal of the American 

Statistical Association 47: 583-621. 

Kruskal, W.H. and Wallis, W.A. 1953. Errata for Kruskal-Wallis 

(1952). Journal of the American Statistical Association 48: 

907-911. 

Kruskal, W.H. 1957. Historical notes on the Wilcoxon unpaired 

two-sample test. Journal of the American Statistical 

Association 52: 356-360. 

SAS/STAT User's Guide, 1989a. SAS Institute Inc., Version 6, 

Fourth Edition, Cary, NC:SAS Institute Inc., 1989. Vol. 1, 

p. 209-244. Vol. 2 p. 1195-1210. 

Page 761
10

HLS STUDY NO.: 00-4204 

TABLE 1 

GASOLINE ETBE VAPOR CONDENSATE: 

A ONE-GENERATION WHOLE-BODY INHALATION REPRODUCTION TOXICITY STUDY IN RATS 

SUMMARY OF MALE REPRODUCTIVE PARAMETERS 

Group: I IV 

Exposure Level: GETBEVCa

 (mg/m3) 0 20,000 

MOTILITY (%) 

 Mean 96 94 

 SD 3 5 

 N 25 26 

EPIDIDYMAL COUNT 

 (MILLION SPERM/GRAM) 

 Mean 956.7 1053.0 

 SD 164.8 215.1 

 N 26 26 

TESTICULAR COUNT 

 (MILLION SPERM/GRAM) 

 Mean 105.7 106.5 

 SD 21.3 21.0 

 N 26 26 

SPERM MORPHOLOGYb

 (% ABNORMAL) 

 Mean 0.8 0.6 

 SD 0.8 0.6 

 N 26 26 

CAUDAL EPIDIDYMIS WEIGHTS 

 (GRAM) 

 Mean 0.27 0.28 

 SD 0.03 0.03 

 N 26 26 

a

GASOLINE ETBE VAPOR CONDENSATE. b

MEAN AND STANDARD DEVIATIONS WERE CALCULATED USING THE TOTAL NUMBER OF ABNORMAL SPERM 

AS A PERCENTAGE OF THE NUMBER OF SPERM EXAMINED. 

NONE SIGNIFICANTLY DIFFERENT FROM CONTROL. 

Page 762
11

HLS STUDY NO.: 00-4204 

APPENDIX A 

GASOLINE ETBE VAPOR CONDENSATE: 

A ONE-GENERATION WHOLE-BODY INHALATION REPRODUCTION TOXICITY STUDY IN RATS 

INDIVIDUAL CAUDAL EPIDIDYMIS WEIGHT, 

SPERM MOTILITY AND SPERM COUNT DATA 

Group I: 0 mg/m3 (GASOLINE ETBE VAPOR CONDENSATE) 

 S P E R M C O U N T 

Animal Caudal Motility Caudal Homogenization Resistant 

 No. Epididymis Epididymal Testicular 

 Weight (g) (%) (million sperm/gram) 

1051 0.28 98 870.1 88.1 

1052 0.24 91 954.1 98.3 

1053 0.23 93 1227.5 91.9 

1054 0.27 100 910.1 114.1 

1055 0.20 97 705.8 105.3 

1056 0.23 98 1036.5 146.9 

1057 0.28 100 780.5 151.1 

1058 0.26 96 949.1 69.7 

1059 0.29 95 739.2 79.6 

1060 0.29 99 1016.8 79.1 

1061 0.25 93 803.0 91.1 

1062 0.28 90 690.9 140.4 

1063 0.34 93 925.7 75.2 

1064 0.26 99 1089.9 107.3 

1065 0.25b a 949.4b 106.9 

1066 0.27 95 844.3 117.7 

1067 0.27 99 1192.8 139.4 

1068 0.27 97 1045.6 109.0 

1069 0.32 92 1035.8 103.9 

1070 0.28 87 1041.9 97.0 

1071 0.28 99 1120.3 111.7 

1072 0.25 97 1183.7 119.0 

1073 0.24 96 1172.0 99.5 

1074 0.27 99 619.6 97.1 

1075 0.27 96 1014.7 115.0 

1076 0.23 92 954.7 94.3 

a

NO SPERM PRESENT FOR ANALYSIS. 

b

LEFT EPIDIDYMIS AND LEFT TESTIS PRESERVED FOR POSSIBLE HISTOPATHOLOGICAL 

EVALUATION; RIGHT EPIDIDYMIS AND RIGHT TESTIS USED FOR SPERM COUNT 

DETERMINATION. 

Page 763
12

HLS STUDY NO.: 00-4204 

APPENDIX A 

GASOLINE ETBE VAPOR CONDENSATE: 

A ONE-GENERATION WHOLE-BODY INHALATION REPRODUCTION TOXICITY STUDY IN RATS 

INDIVIDUAL CAUDAL EPIDIDYMIS WEIGHT, 

SPERM MOTILITY AND SPERM COUNT DATA 

Group IV: 20,000 mg/m3 (GASOLINE ETBE VAPOR CONDENSATE) 

 S P E R M C O U N T 

Animal Caudal Motility Caudal Homogenization Resistant 

 No. Epididymis Epididymal Testicular 

 Weight (g) (%) (million sperm/gram) 

4051 0.28 94 1105.4 101.0 

4052 0.30 97 962.0 88.4 

4053 0.29 94 1305.2 76.9 

4054 0.23 97 1272.9 94.2 

4055 0.19 79 627.4 77.8 

4056 0.30 99 930.6 82.7 

4057 0.26 88 1351.3 120.5 

4058 0.25 94 819.8 97.8 

4059 0.28 88 903.7 93.3 

4060 0.32 98 1195.9 93.2 

4061 0.28 96 1198.7 92.2 

4062 0.28 97 1333.2 122.6 

4063 0.23 96 836.5 144.0 

4064 0.30 89 1153.7 77.0 

4065 0.21 95 1050.6 110.2 

4066 0.28 97 1056.8 105.3 

4067 0.27 93 867.5 128.7 

4068 0.32 97 1392.0 87.8 

4069 0.29 96 1117.7 108.7 

4070 0.25 95 1292.4 144.3 

4071 0.30 98 1104.9 105.2 

4072 0.30 87 1164.1 126.2 

4073 0.32 98 630.6 108.4 

4074 0.28 94 847.7 121.0 

4075 0.29 100 1020.4 151.0 

4076 0.25 88 836.5 110.1 

Page 764
HLS STUDY NO.: 00-4204 

APPENDIX B 

GASOLINE ETBE VAPOR CONDENSATE: 

A ONE-GENERATION WHOLE-BODY INHALATION REPRODUCTION TOXICITY STUDY IN RATS 

INDIVIDUAL SPERM MORPHOLOGY DATA 

13

Group I: 0 mg/m3 (GASOLINE ETBE VAPOR CONDENSATE) Animal ----------------H e a d---------------- --------T a i l---- No. Normal Amorphous Small Enlarged Double Coiled Bent D

1051 200 0 0 0 0 0 0 1052 200 0 0 0 0 0 0 1053 199 1 0 0 0 0 0 1054 196 4 0 0 0 0 0 1055 198 2 0 0 0 0 0 1056 198 2 0 0 0 0 0 1057 200 0 0 0 0 0 0 1058 199 1 0 0 0 0 0 1059 200 0 0 0 0 0 0 1060 197 3 0 0 0 0 0 1061 196 3 0 1 0 0 0 1062 199 1 0 0 0 0 0 1063 198 1 1 0 0 0 0 1064 197 3 0 0 0 0 0 1065 196 4 0 0 0 0 0 1066 198 2 0 0 0 0 0 1067 198 1 0 0 0 1 0 1068 200 0 0 0 0 0 0 1069 200 0 0 0 0 0 0 1070 200 0 0 0 0 0 0 1071 199 1 0 0 0 0 0 1072 200 0 0 0 0 0 0 1073 200 0 0 0 0 0 0 1074 198 2 0 0 0 0 0 1075 200 0 0 0 0 0 0 1076 195 5 0 0 0 0 0 
HLS STUDY NO.: 00-4204 

APPENDIX B 

GASOLINE ETBE VAPOR CONDENSATE: 

A ONE-GENERATION WHOLE-BODY INHALATION REPRODUCTION TOXICITY STUDY IN RATS 

INDIVIDUAL SPERM MORPHOLOGY DATA 

14

Group IV: 20,000 mg/m3 (GASOLINE ETBE VAPOR CONDENSATE) Animal ----------------H e a d---------------- --------T a i l---- No. Normal Amorphous Small Enlarged Double Coiled Bent D

4051 200 0 0 0 0 0 0 4052 199 1 0 0 0 0 0 4053 198 2 0 0 0 0 0 4054 200 0 0 0 0 0 0 4055 198 2 0 0 0 0 0 4056 198 1 0 0 0 1 0 4057 200 0 0 0 0 0 0 4058 200 0 0 0 0 0 0 4059 199 1 0 0 0 0 0 4060 200 0 0 0 0 0 0 4061 198 2 0 0 0 0 0 4062 200 0 0 0 0 0 0 4063 198 2 0 0 0 0 0 4064 200 0 0 0 0 0 0 4065 195 5 0 0 0 0 0 4066 198 2 0 0 0 0 0 4067 198 2 0 0 0 0 0 4068 200 0 0 0 0 0 0 4069 200 0 0 0 0 0 0 4070 199 1 0 0 0 0 0 4071 198 2 0 0 0 0 0 4072 200 0 0 0 0 0 0 4073 197 3 0 0 0 0 0 4074 200 0 0 0 0 0 0 4075 199 1 0 0 0 0 0 4076 198 2 0 0 0 0 0 
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Certificates of Analysis Appendix CC 

1. Feed Certificates of Analysis ....................................................................... 768 

2. Water Certificates of Analysis ..................................................................... 794 

3. Bedding Certificates of Analysis .................................................................. 801 
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Protocol and Protocol Amendments Appendix DD 
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Testing Facility Personnel Appendix EE 

TITLE/DEPARTMENT NAME/DEGREE 

SENIOR VICE PRESIDENT, 

 SAFETY ASSESSMENT 

Sylvie J. Gosselin, D.V.M., Ph.D., 

 Diplomate A.C.V.P. 

 

SENIOR DIRECTOR, SAFETY 

ASSESSMENT 

Carol S. Auletta, M.B.A., D.A.B.T., 

 R.A.C. 

 

DIRECTOR, ANALYTICAL SERVICES Barbara A. Litzenberger, B.S., 

 M.T. (ASCP) 

 

DIRECTOR, TOXICOLOGY 

 OPERATIONS Ian Vanterpool, F.I.A.T. 

 

DIRECTOR, QUALITY ASSURANCE Nicki S. Iacono, B.S. 

STUDY DIRECTOR Gary M. Hoffman, B.A., D.A.B.T. 

PATHOLOGIST Katharine M. Whitney, D.V.M., 

 Ph.D., Diplomate A.C.V.P. 

 

STUDY MONITOR Janet E. Scimone, B.A. 

VETERINARIAN Teresa S. Kusznir, V.M.D. 

MANAGER/SUPERVISOR 

 Reproductive Toxicology 

Inhalation 

 Analytical Services 

 Clinical Pathology 

 Histology 

Robert Faust, B.S. 

Ellen H. Whiting, AALAS LAT 

Kay Saladdin, B.S. 

Lisa A. Spall, B.S., M.T. (ASCP) 

Janet Kusisto 
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Report Amendments Appendix FF 

There are no amendments for this report at this time.