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### Working for a better future



## REPORT 2 - bromo - 3,3,3 -trifluoropropene : Reproductive/ Deve lopment al Toxicity Screening Study in the CD Rat by Inhalation

Administration

HLS s tudy number: WAG0015 Version ID: Final Issue date: 22 July 2013Huntingdon Life Sciences WAG0015

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# Details of Sponsor and Te st Facilit ies S ponsor American Pacific Corporation 3883 Howard Hughes Parkway Suite 700 Las Vegas Nevada 89169 USA Test facility Huntingdon Life Sciences Primary loc ation Huntingdon Research Centre Wo olley Road Alconbury Huntingdon Cambridgeshir

e PE28 4HS UK Test facility Huntingdon Life Sciences Oestrous cycles , Eye Research Centre Sperm analysis Eye Histology, Suffolk Histopathology IP23 7PX UKHuntingdon Life Sciences W AG0015

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# Ta ble of Contents Details of Spo nsor and Test Facilities ................................................................................... 2 Compliance with Good Laboratory Practice .......................................................................... 8 Quality Assurance Statement ................................................................................................ 9 Contributing Scientists ........................................................................................................ 10 Summary ............................................................................................................................ 11 1. Introduction ................................................................................................................ 14 1.1 Objective ......................................................................................................... 14 1.2 Regulatory compliance ................................................................................... 14 1.3 Test system .................................................................................................... 14 1.4 Route of adminis tration ................................................................................... 14 1.5 Treatment groups and doses .......................................................................... 14 2. Experimental Procedure ............................................................................................. 15 2.1 Study schedule and structure .......................................................................... 15 2.1.1 Duration of treatmen t ........................................................................... 15 2.1.2 Time schedule ..................................................................................... 15 2.1.3 Study design ...................................................................................... 15 2.1.4 Identity of treatment groups ................................................................ . 16 2.2 Test substance and formulation ...................................................................... 16 2.2.1 Test substance .................................................................................... 16 2.2.2 Control ................................................................................................ 17 2.3 Animal management ....................................................................................... 17 2.3.1 Animal supply, acclimatisation and allocation ...................................... 17 2.3.2 Animal housing, diet and water supply ................................................ 17 2.3.3 Administration ..................................................................................... 19 2.4 Serial observations ......................................................................................... 19 2.4.1 Clinical observations ........................................................................... 19 2 .4.2 Mortality .............................................................................................. 20 2.4.3 Bodyweight ......................................................................................... 20 2.4.4 Food consumption ............................................................................... 20 2.4.5 Wa ter consumption ............................................................................. 20 2.4.6 Oestrous cycles ................................................................................... 21 2.4.7 Mating procedure ................................................................................ 21 2.4.8 Parturition observations and gestation length ...................................... 21 2.4.9 Records made during littering phase ................................................... 21 2.5 Necropsy and histology ................................................................................... 22 2.5.1 Time of necropsy ................................................................................. 22 2.5.2 Method of kill ....................................................................................... 22 2.5.3 Macroscopic pathology ........................................................................ 22 2.5.4 Sperm analysis .................................................................................... 23 2 .5.5 Organ weights ..................................................................................... 24 2.5.6 Fixation ............................................................................................... 25 2.5.7 Histology ............................................................................................. 25 2.6 Pathology ........................................................................................................ 26 2.6.1 Light microscopy ................................................................................. 26 2.7 Computer systems .......................................................................................... 26 2.8 Data treatment ................................................................................................ 27 2.8.1 Signs ................................................................................................... 28Huntingdon Life Sciences WAG0015

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### 2.8.2 Bodyweights ........................................................................................ 28 2.8.3 Food consumption ............................................................................... 28 2. 8.4 Wa ter consumption ............................................................................. 28 2.8.5 Oestrous cycles ................................................................................... 29 2.8.6 Pre- coital interval ................................................................................ 29 2.8.7 Mating performance and fertility .......................................................... 29 2.8.8 Gestation length .................................................................................. 29 2.8.9 Litter size ............................................................................................. 29 2.8.10 Survival indices ................................................................................... 30 2.8.11 Sex ratio .............................................................................................. 30 2.8.12 Sperm analysis .................................................................................... 30 2. 8.13 Organ weights ..................................................................................... 30 2.8.14 Pathology ............................................................................................ 31 2.8.15 Statistical analysis ............................................................................... 31 2.9 Quality assurance and archiving procedures .................................................. 33 2.9.1 Quality assurance ............................................................................... 33 2.9.2 Archives .............................................................................................. 33 2.10 Deviations from protocol ................................................................................. 34 3. Results ....................................................................................................................... 35 3.1 Chamber atmosphere conditions .................................................................... 35 3.2 F0 maternal responses ................................................................................... 35 3.2.1 Mortality and animal fates .................................................................... 35 3.2.2 Signs ................................................................................................... 35 3.2.3 Bodyweight ......................................................................................... 36 3.2.4 Food consumption ............................................................................... 37 3.2.5 Wa ter consumption ............................................................................. 37 3.2.6 Oestrous cycle length .......................................................................... 37 3.2.7 Pre- coital interval, mating performance and fertility ............................. 38 3.2.8 Gestation length and gestation index .................................................. 39 3.2.9 Sperm analysis .................................................................................... 39 3.2.10 Organ weights ..................................................................................... 40 3.2.11 Macropathology ................................................................................... 40 3.2.12 Animals killed after sche duled treatment period .................................. 40 3.2.13 Histopathology .................................................................................... 41 3.2.14 Animals killed after scheduled treatment period .................................. 41 3.2.15 Primordial ovarian follicle counts ......................................................... 43 3.3 F1 litter responses .......................................................................................... 43 3.3.1 Signs of offspring ................................................................................ 43 3.3.2 Litter size, survival indices and sex ratio .............................................. 43 3.3.3 Bodyweight ......................................................................................... 44 3.3.4 Offspring macropathology ................................................................... 44 4. Discussion .................................................................................................................. 45 5. Conclusion ................................................................................................................. 48 6. References ................................................................................................................. 49 6.1 Necropsy and histology ................................................................................... 49 6.2 Statistics ......................................................................................................... 49Huntingdon Life Sciences WAG0015

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# List of Figures Figure 1 Bodyweight - group mean values for males .................................................. 50 Figure 2 Bodyweight - group mean values for females before pairing ........................ 51 Figure 3 Bodyweight - group mean values for females during gestation ..................... 52 Figure 4 Bod yweight - group mean values for females during lactation ...................... 53 Figure 5 Bodyweight - group mean values for offspring ............................................. 54 

# List of T ables Table 1 Summary of adult performance .................................................................... 56 Table 2 Clinical signs - group distribution of observations ......................................... 57 Table 3 Signs associated with dosing - group distribu tion of observations ................ 62 Table 4 Bodyweight and bodyweight change - group mean values for males ........... 77 Table 5 Bodyweight and bodyweight change - group mean values for females before

pairing .......................................................................................................... 79 Table 6 Bodyweight and bodyweight change - group mean values for females during

gestation ...................................................................................................... 81 Table 7 Bodyweight and bodyweight change - group mean values for females during

lactation ....................................................................................................... 83 Table 8 Food consumption - group mean values before pairing ................................ 84 Table 9 Food consumption - group mean values for females during gestation .......... 86 Table 10 Food consumption - group mean values for fema les during lactation ........... 87 Table 11 Wa ter consumption - group mean values for males and before pairing for

females ........................................................................................................ 88 Table 12 Wa ter consumption - group mean values for females during gestation ........ 90 Table 13 Wa ter consumption - group mean values for females during lactation .......... 92 Table 14 Oestrous cycles - group values .................................................................... 93 Table 15 Pre- coital interval - group values .................................................................. 95 Table 16 Number of copulation plugs at mating - group values ................................... 96 Table 17 Sperm count estimates from vaginal smears at mating - group values ......... 97 Table 18 Mating performance and fertility - group values ............................................ 98 Table 19 Gestation length a nd gestation index - group values .................................... 99 Table 20 Litter size - group mean values .................................................................. 100 Table 21 Offspring survival indices - group mean values .......................................... 101 Table 22 Sex ratio - group mean values ................................................................... 102 Table 23 Bodyweight and bodyweight change - group mean values for

male offspring ............................................................................................ 103Huntingdon Life Sciences WAG0015

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### Table 24 Bodyweight and bodywei ght change - group mean values for

female offspring ......................................................................................... 104 Table 25 Sperm analysis - group mean values ......................................................... 105 Table 26 Sperm motion data - group mean values ................................................... 106 Table 27 Sperm morphology - group mean values ................................................... 107 Table 28 Organ weights - group mean unadjusted and adjusted values for males .... 108 Table 29 Macrop athology - group distribution of findings .......................................... 109 Table 30 Histopathology - group distribution of findings ............................................ 115 Table 31 Primordial ovarian follicle counts - group mean values ............................... 119 

# List of Appendices Appendix 1 Clinical signs - individual observations for males ........................................ 120 Appendix 2 Clinical signs - individ ual observations for females ..................................... 123 Appendix 3 Signs associated with dosing - individual observations for males

and females ............................................................................................... 134 Appendix 4 Signs associated with dosing - individual observations for females prior to

mating, during gestation and lactation ........................................................ 194 Appendix 5 Bodyweight - Individu al values for males ................................................... 235 Appendix 6 Bodyweight - individual values for females before pairing .......................... 239 Appendix 7 Bodyweight - individual values for females after mating .............................. 242 Appendix 8 Bodyweight - individual values for females during lactation ........................ 244 Appendix 9 Food consumption - individual values for males and before pairing

for females ................................................................................................ . 246 Appendix 10 Food consumption - individual v alues for females after mating ................... 248 Appendix 11 Food consumption - individual values for females during lactation ............. 250 Appendix 12 Wa ter consumption - individual values for males before and after pairing an d female s before mating - F0 generation (mL/animal/day) ........................................ 252 Appendix 13 Wa ter consumption - individual values for females after mating ................. 256 Appendix 14 Wa ter consumption - individual values for females during lactation ............ 260 Appendix 15 Oestrous cycle length, mating performance, fertility and gestation length -

individual values ........................................................................................ . 262 Appendix 16 Litter size - individual values ...................................................................... 266 Appendix 17 Offspring survival indices - individual values .............................................. 270 Appendix 18 Sex ratio - individual values ....................................................................... 274 Appendix 19 Bodyweight - individual litter mean values for offspring .............................. 278 Appendix 20 Clinical signs - individual findings for offspring ........................................... 285 Appendix 21 Macropathology - individual findings for offspring dying or killed before

scheduled termination ................................................................................ 289Huntingdon Life Sciences WAG0015

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### Appendix 22 Macropathology - individual findings for offspring killed at scheduled

termination ................................................................................................ . 291 Appendix 23 Sperm analysis - individual values .............................................................. 292 Appendix 24 Sperm motion data - individual values ........................................................ 296 Appendix 25 Sperm morphology - individual values ........................................................ 300 Appendix 26 Organ weights - individual absolute values for males ................................ . 304 Appendix 27 Macropathology and histopathology - individual findings ............................ 306 Appendix 28 Primordial ovarian follicle counts ................................................................ 386 

# List of Annexes Annex 1 Certificate of an alysis ................................................................................. 394 Annex 2 Aerosol technology and inhalation analysis report ...................................... 396 Annex 3 Pathology report ......................................................................................... 415 Annex 4 GLP compliance statement s ...................................................................... 425Huntingdon Life Sciences WAG0015

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# Contributing Scientist s 2-bromo- 3,3,3 -trifluoropropene: Reproductive/Developmental Toxicity Screening Study in the CD Rat by Inhalation Administration 

## Study management Amanda J Brooker BSc (Hons) MSc CBio l MSB Study Di rector David P Myers BSc (Hons) PhD IDT Senior Toxico lo gist Steven J Crome BSc MSc Head of Toxico lo gists (inhalation aspects) 

## Formulation analysis Caroline Finucane PhD Head of Form

ulat io n & Inhalat io n Analysis 

## Aerosol technology Simo n A Moore BSc (Hons) PhD MRSC Head of Aerosol Techno lo gy 

## Oestrous cycles and m ating performance Martin J Co llier BSc Behavioural Scient ist 

## Sperm analysis Clare Bowden BSc (Hons) CBio l MSB Operational

Head, Sperm

Analysis & Embryo Cul

ture 

## Pathology Miledi Azzalin DVM MRCVS Pathologist Department of Pathology 

## Statistics Gareth D Thomas BSc (Hons) Head of Statist ic s and Data ManagementHuntingdon Life Sciences WAG0015

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# Su mmary The obj ect ive o f this study was an init ial screening assessment of the influence of 2- bromo- 3,3,3 - trifluoropropene (a fire ext inguisher agent ), on reproductive performance and development in Crl :CD (SD) rat s fo llo wing inhalatio n administration. Thre e groups of ten male and ten fe male rats received 2- bro mo - 3,3,3 - trifluoropropene by inhalat io n at target exposure levels of 200, 500 or 3000 ppm, 7 days per week, 6 hours per day . The adults were tr eated daily for a minimum o f 15 days before pairing unt il Day 9 af ter the birth of the F1 generat io n , and were sacrificed on Day 10 af ter the bi rth of the F1 generation . Females were untreated fro m Day

20 af ter mating unt il Day 4 after birth inclusive . A similarly constituted Control group received ai

r only fo r the same duration . The F1 generat io n received no direct administration of the test substance; any exposure was in utero or via the milk. During the study, clinical condit io n, bodyweight, food consumpt io n, macroscopic and microscopic patho lo gy

invest igations were undertaken in all adults. Oestrous cycles and g estation length were assessed and parturi tion observat io ns were performed for F0 females. Organ weights were recorded for F0 males only. The clinical condit io n, litter size and survival, sex ratio and body weight of all o ffspring were assessed. 

## Results The achieved chamber concentrations o f 2 bromo- 3,3,3 - trifluoropropene were 198, 505 and 2900 ppm (99, 101 and 97% of the target concentration) for Groups 2, 3 and 4 respectively. At2900 ppm, treatme ntrelated findings included clinical signs during exposure of underact ivit y, unresponsiveness, piloerection and partially clos ed eyelids, occasionally in females these lasted aft er com plet io n of exposure, and included hunched posture, but not at the end of the day; pale teeth in all males and the majorit y of f emales and slight ly higher incidence of hairloss in males (confirmed at macroscopic examinat io n) ; lo wer body weight gain of males throughout and females prior to pairing and during gestation; lo wer f ood int ake; increased water intake for females; longer oestrous cy cles (6 days or longer) wi th mo re f emales showing i

rregular cycles or being acyclic; reduced fert ilit y ( 6 /10 pai red f emales were pregnant); longer pre - co it al int erval; at mating, fewer copul ation plugs and lower estimates of sperm in the vaginal smear; extended duration of gestation with only 1 female littering on Day

25 of gestation and 2 females being sacrificed on Day 24 of gestation); lower implantation counts, wi

th only one f emale litter ing and the offspring subsequent ly dying; effects on sperm analysis (reduced percentage progressively mot ile sperm, sperm velo ci ty, increased BCF, reduced sperm count in the cauda epididymis, increase in abnormal sperm – breakages and abnormal head shape); reduced prostate, epididymides, seminal vesicle and pi tui tary weights; small prostate macroscopically in all males ; in the majorit y of males and some females, spleen capsular thickening and adhesions noted macroscopically and confirmed microscopically as capsular/subcapsular inflammat io n/capsular thickening and /or adhesio ns/infla mmat io n/ fibrosis and reduced size of corpora lutea in the ovaries. At505 ppm, treatment related findings included clinical signs during exposure of underact ivit y and piloerection and occasio nally unresponsiveness and partially clos ed eyelids, occasionally

in females so me of these la st ed aft er complet io n of exposure but not at the endHuntingdon Life Sciences WAG0015

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### of the day; pale teeth in some ma les and females in the last week of treatment; lo wer body weight g ain of males throughout and slight ly lower bodyweight gain of females during gestation; slight bodyweight loss for females in lactation; lower food intake; increased water int ake for females prior to pai ring and during gestation, with lower water intake during lactation ; lo nger oestrous cy cles (6 days or longer) in 5/10 females wi th more females showing irregular cycles or being acyclic; slight ly lo nger pre- coi tal int erval; at mat ing, fewer copulat io n pl

ugs and lower estimates of sperm in the vaginal smear; extended duration of gestation (23 - 25 days) ; slightly lo wer implantation counts; lower post im plantation survival , live birth index and live litter size on Day 1; clinical

signs in o ffspring of cold to touch, unfed and reduced act ivit y necessitat ing sacri fice along with an increased incidence of dead offspring result ing in only 4/9 litters surviving to Day 10 post partum; effects on sperm analysis (reduced sperm velocit y, increase in abnormal sperm

– breakages and abnormal head shape); reduced prostate, ep id idymides, seminal vesicle and pituitary weights; small prostate macroscopically in the maj ori ty of males ; and spleen capsular thickening and adhesio ns noted macroscopically in the maj ori ty of males and occasional females and confirmed microscopically as capsular/subcapsular inflammatio n/capsular thickening and /or adhesio ns/infla mmat io n/ fibrosis. At198 ppm, slight effects on clinical signs during exposure, bodyweight, food and water int ake were considered not adverse. Of uncertain significance was a sli ghtly lower sperm count noted in the vaginal smear on the day of mating, but not statist ic ally significant ly affected at termination, and a shift to a 23 day duration of gestation, and an increase in abnormal sperm . The mean number of implantations was slight ly lo wer than the Controls. Three out of 9 litters showed lower than expected post implantation survival of the conceptuses (63 - 74%) and two litters showed lower than expected pup viabilit y indices (54 and 82%), and these differences contributed to lower total and live li

tter sizes than the Controls. Alt hough only a few litters were invo lved, the reduction in total and live litter size on Day 1 of lactation f

ormed part of a dose related trend and thus a relat io nshi p to treatment was considered likely. 

## Co nclusion Exposure of 2 - bro mo -3,3,3- trifluoropropene to rats in this screening study to assess the influence on reproductive performance and develo pment f ollo wing administration by inhalat io n at concentrations of 198, 505 or 2900 ppm was associated with adverse effects on male and female toxicit y, and reproductive performance and development at 505 and 2900 ppm . Exposure at 505 and 2900 ppm produced effects on oestrous cycles, fert ilit y (2900 ppm only), extended duration of gestation, sperm counts and motili ty, implantation counts, reduced in utero and post partum survival, in the presence of signs of adult toxicit y. In addi tion, at 2900 ppm, reduced size of corpora l utea in the ovaries was noted. Exposure at 198 ppm was associated with slight effects on ad ulttoxici ty, but did have slight effects on reproductive performance (a possible shift to longer duration of gestation) and development (slightly lower implantation rate, higher post implantation survival and viabilit y indices leading to lower litter size ) as seen at 505 ppm and above.Huntingdon Life Sciences WAG0015

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### Hi stopathological

examinat io n of the reproductive organs, including the testes and epididymides, revealed no findings attributable to exposure to the test article, 2 - bromo- 3,3,3 - trifluoropropene. Wit hin this screening study, the no adverse effect level (NOAEL) for reproductive performance and development was considered to lie below 198 ppm .Huntingdon Life Sciences WAG0015

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# 1. Introduction 1.1 Objective The obj ect ive o f this study was an init ial screening assessment of the influence of 2- bromo- 3,3,3 - trifluoropro pene , an experimental agent under development to replace halo ns as a fire ext inguishing agent , on reproductive performance and development fo llo wing administration by inhalat io n fo r a minimum of 15 days before pairing unt il Day 9 af ter the birth of the F1 generat io n wi th sacrifice on Day

10 aft er the birth of the F1 generat io n. 

## 1.2 Regulatory compliance The study was designed to meet the requirements of the fo llo wing guidelines: Organisat io n fo r Economic Co - operation and Development: Testing of Chemicals (Guide line 421; Reproduction/Developmental Toxic it y Screening Test; 27 July 1994 ). The study was conducted in accordance wit h the requirements of current, internat io nally recognised Good Laboratory Practice Standards, and the applicable sect io ns of the United Kingdom Animals (Scient ific Procedures) Act 1986. 

## 1.3 Test system The rat was chosen as the test species because of the requirement for a rodent species by regulat ory

agencies. The Crl :CD (SD) strain was used because of the historical

control data available at this laboratory . 

## 1.4 Route of administration The inhalat io n route of administration was chosen to simulate the condit io ns of potential human exposure . 

## 1.5 Treatment groups and doses The exposure levels were based on results of 2 week dose range finding studies in rats performed in these laboratories ( Huntingdon Life Sciences Report Numbers WAG0013 and WAG0024 respectively). In those studies, clinical signs were noted during exposure at target exposure levels o f 500 ppm to 20000 ppm, wi

th excessive body

weight effects at 10000 ppm and above. Respiratory

tract pathology

was noted at target exposure levels o f 1200 ppm and above. It was considered suitable for the target exposure levels o f this reproduct ive study to replicate that of the 90 day

rat study ( Hunt ingdon Life Sciences Report Number WAG0014 ). The high exposure concentration, 3000 ppm, was selected to allow assessment of reproductive effects at an exposure concentration ant ic ipated to produce evidence of systemic toxicit y. Lower exposure concentrations were chosen to allow assessment of any effects observed.Huntingdon Life Sciences WAG0015

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# 2. Experimental Procedure 2.1 Study schedule and structure 2.1.1 Duration of treatment The test substance, 2- bro mo -3,3,3- trifluoropropene, was administered to F0 adults for a minimum o f 15 days before pai ring unt il Day 9 af

ter the bi rth of the F1 generat io n . The F0 female adults were killed on Day 10 af ter the bi rth of the F1 generat io n and the F 0 ma les killed after the start of the F0 female kills . F0 females were untreated fro m Day 20 af ter mat ing to Day 4 after birth. Animals o f the F1 generat io n were not dosed, but may have been exposed to the test substance or metabo lit es in utero or through the milk . 2.1.2 Time sched ule Study init iat io n: (Pro tocol signed by

Study Di rector) 09 July

2012 Experimental start date: ( Animal arrival) 12 July 2012 Treatment commenced: 19 July 2012 Pai ring co mmenced: 02 August 2012 Necropsy com pleted : F0 Males F0 Females and F1 animals 07 September 2012 13 September 2012 Experimental co mp let io n date: (Pathology

) 13 November 2012 Study com plet io n: When the Study Di rector signs the final report 2.1.3 Study design Sacrifice

Males

Females Mating

Male treatment period Female treatment period 15 days before pairing Day 10 of lactatio n

Dams not exposed Day 20 post coitum to Day 4 post - partum inclusiveHuntingdon Life Sciences WAG0015

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### 2.1.4 Identity of treatment groups The study consisted of one Control and three treated groups of rats, ident ified as fo llo ws: Group Treatment Target exposure level (ppm) Number of animals Animal numbers

Male Female Male Female 1 Control 0 10 10 20 1 - 2 10 2 41 - 2 50 2 2 - br omo - 3,3,3 -

trifluoropropene 200 10 10 2 11 - 2 20 2 51 - 2 60 3 2 - br omo - 3,3,3 -

trifluoropropene 500 10 10 2 21 - 2 30 2 61 - 2 70 4 2 - br omo - 3,3,3 -

trifluoropropene 3000 10 10 2 31 - 2 40 2 71 - 2 80 

### The F1 generat io n received no direct administration of the test substance, 2- bromo- 3,3,3 - trifluoropropene . Any exposure to the test substance or metabolites was th rough the mother to the offspring in utero and/or through the milk , or f rom transfer from the mother’s fur fo llo wing exposure from Day 5 after bi rth. 

## 2.2 Test substance and formulation 2.2.1 Test substance Informat io n supplied by the Sponsor regarding the test substance is contained in the test substance data sheet, which is retained in study records, and the Certificate of Analysis, which is presented in Annex 1. The fo llo wing in fo rmat io n is given in summary: Ident ification: 2- bromo- 3,3,3 - trifluoropropene (stabilized) Act io n: Fi

re ext inguisher agent Descript io n: Clear liquid wi th a slight yellow hint Storage condit io ns: Atambient temperature Supplier: Sponsor Batch number: WN CC - BTP111128 D ate of receipt: 02 March 2012 Expi ry date: 12 January 201 3 Puri ty : 99.9% excluding stabilisersHuntingdon Life Sciences WAG0015

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### Stabilisers:

The Sponsor was responsible for the characterisat ion of the test substance and the documentation of

the methods of synt hesis, fabrication or derivat io n and stabilit y. 2.2.2 Control Group 1 (Control) animals were exposed to air only. 

## 2.3 Animal management 2.3.1 Anima l supply, acclimatisation and allocation Crl :CD (SD) rats (a total of 45 ma les and 45 females) were received f rom Charles River (UK) Ltd . They were delivered at approximately 49 to 56 days of age and wi thin a weight range of 250 to 3 00 g for males and 2 00 to 2 30 g for females. On arrival, the animals were removed fro m the transit boxes and allocated to study cages. Using the sequence of cages in the battery, one animal at a time was placed in each cage wit h the procedure being repeated until each cage held the appropriate number of animals. Each sex was allo cated separately. The day af ter arrival a representative sample of animals were weighed and thei r clinical condi tion was sat is factory ; based on this the animal s were deemed accepta ble for use on the study . On Day

1 (bef ore dosing) variat io ns in group bodyweights were confirmed to be wi thin ± 20% of the mean weight fo r each sex. Each adult animal was assigned a number and identified uniquely within the study by a tail ta ttoo. Each cage label was co lo ur- co ded according to group and was numbered uniquely wi th cage and study

n umber, as well as the ident it y of the occupant(s). The adult animals were allowed to acclimat ise to the condit io ns described below fo r 7 days before treatment commenced. For those animals selected for this study, thei r age at the start of treatment was 56 to 63 days and their body weights were in the range o f 303 to 349 g for males and 204 to 247 g for fe males . The offspring were numbered individually wi thin each litter on Day

1 of age, using a toe ta ttoo. The spare animals were remo ved from the study room af ter treatment commenced. 2.3.2 Animal housing, diet and water supply Animals were housed inside a restricted access rodent facilit y (Building Y14 , Room 010) . The facilit y was designed and operated to minimise the entry of external biological andHuntingdon Life Sciences WAG0015

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### chemical agents and to minimise the transference of such agents between rooms. Before the study the room was cleaned and disinfected. Each animal room was kept at positive pressure with respect to the outside by its own supply of f iltered fresh air, which was passed to atmosphere and not re - circulated. The temperature and relative humidit y controls were maintained wit hin the range of 19 to 2 5  C and 40 to 70% respectively . Alt hough condit io ns were occasionally outside the indicated ranges, these deviat io ns were minor and/or of short duration and were not considered to have influenced the healt h of the animals and/or the outcome of the study. Artificia l light ing was controlled to give a cycle o f 12 hours continuous light and 12 hours continuous dark per 24 hours, with the lights on at 6:00 GMT. Alarms were activated if there was any failure of the vent ilat io n system, or temperature limits were exceeded. A stand - by electrici ty supply was availab le to be automat ically bought into operation should the public supply fail. The animals were housed as shown below: Study period Number of animals/cage Cage material Cage flooring Male Female Pre - pairing 5 5 Pol ycarbonate Solid polycarbonate P airing 1 : 1 Pol ycarbonate Stainless steel grid Males after mating Up to 5 - Pol ycarbonate Solid polycarbonate Gestation - 1 Pol ycarbonate Solid polycarbonate Lactatio n - 1 (+ litter) Pol ycarbonate Solid polycarbonate 

### The cages were distributed on the racking to equalise, as far as possible, environmental influences amo ngst the groups. Wood based bedding was used in cages wit h solid floors and was sterilised by autoclaving and changed at appropriate intervals each week. The gridded cages were suspended over trays covered with absorbent paper which was changed daily during mat ing. IMS Nestlets were provided from Day 15 after mat ing and throughout parturit io n and lactation. Cages, cage - trays, f ood hoppers and water bottles were changed at appropriate intervals. Each animal was provided with an Aspen chew block for environmental enrichment. Chew blocks were provided throughout the study (except during pai

ring and la ctation), and were replaced when necessary . Each cage of animals was provided with a plastic shel ter f or environmental enrichment (except during pai ring , gestation and lactation), which was replaced at the same time as the cages. The animals were allowed free access to a standard rodent breeding diet (SDS VRF1 Certified diet). This diet is higher in protein and some vitamin content than standard rodent diet and contained no added antibiot ic or other chemotherapeut ic or prophylact ic agent. Potable water taken fro m the public supply was f reely available via polycarbonate or polypropyle ne bottles fitted with sipper tubes. Each batch of diet was analysed routinely by the supplier for various nutrit io nal com ponents and chemical and microbio lo gical contaminants. Supplier's analy ti cal certificates were scrutinised and approved before any b atch of diet was released for use. The qualit y of theHuntingdon Life Sciences WAG0015

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### water supply

i s governed by

regulat io ns published by the Department f

or Envi ronment, Food and Rural Affairs. Certificates of analysis were received routinely fro m the water supplier, the bedding suppli er and the supplier s of the A spen chew blo cks , IMS Nestlets and plast ic shel ters . Since the results of these various analyses did not provide evidence of contaminat io n that might have prejudiced the study, they are not presented. No other specific contaminants that were likely to have been present in the bedding , diet or water were analysed, as none that may have interfered with or prejudiced the outcome of the study was known. 2.3.3 Administration Treatment was restricted to the F0 generat io n. Animals o f the F1 generat io n were not treated di rect ly but may

have been exposed to the test substance or metabolites in utero or through the milk. The test substance was administered to rats using who le body exposure systems. Animals in the treated groups were exposed to aerosols generated from 2 - bromo- 3,3,3 - trifluoropropene at target exposure levels o f 200, 500 and 3000 ppm. Control animals were exposed to air only, using the same dosing procedure. The relevant treatment was administered once daily fo r a period of

6 - hours, seven days per week . Exposure of pregnant females was not undertaken on and after Day 20 af ter mat ing and not reintroduced until Day 4 post partum. The test atmosphere generat io n, administration, sampling and analysis and chamber environment, wa s shared with that of a concurrent 13 week study in rats (WAG0014) with the same test article and target concentrations, on weekdays (Monday to Friday ). Precise details of administration and analysis of the test atmospheres together with the results obtained are presented in Annex 2 . 

## 2.4 Serial observations Dated and signed records of all act ivit ies relating to the day by day running and maintenance of the study wi thin the animal unit as well as to the group observat io ns and examinat io ns outlined in this experimental procedure were recorded in the Study Daybook. In addi

tion, observat io ns relat ing to individual animals made throughout the day were recorded. 2.4.1 Clinical observations Animals were inspected visually at least twice daily for evidence of ill- healt h or r eaction to treatment. Cages and cage - trays were inspected daily f or evidence of ill - healt h amongst the occupant(s). Any

deviat io n fro m normal was recorded at the time in respect of nature and severit y, date and time of onset, duration and progress of the observed condit io n, as appropriate. In addit io n, a more detailed physical examinat io n was performed once each week on each F0 animal (f emales only unt il mat ing ) and for F0 females on Days 0, 7 , 1 4 and 20 after mat ing and Days 1, 5 , and 9 of

lactation to mo nit or general healt h. Daily during the first week of treatment, twice weekly during Weeks 2 to 4 and weekly thereafter for all adult animals (females only until mat ing) and, for females, on Days 0, 7, 14Huntingdon Life Sciences WAG0015

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### and 19 after mat ing and Days 5 and 9 of lactatio n, detailed observat io ns were recorded at the fo llo wing t imes in relat io n to dose administration:

- Pre exposure observation

- Observations during exposure (limit ed to animals visible in the exposure chamber), recorded on a cage basis at approximately 2 and 4 hours after start of exposure

- Im mediately after dosing on return of the animal to its cage

- As late as possible in the working day For the observat io ns during exposure, all animals in each group have been given any clinical signs observed as only a few ani mals could be observed. 2.4.2 Mortality Debilitated animals were observed carefully and animals were killed for reasons of animal welfare where necessary . A com plete necropsy

was performed in all cases for animals killed or found dead, as described in the Necro psy section below. 2.4.3 Bodyweight The weight of each adult was recorded during acclimat isat io n, on the day that treatment commenced (Week 0), twice weekly thereafter (females only unt il mating) and before necropsy . The weight of each F0 female was also recorded on Days 0, 3, 7, 10, 14, 17 and 20 after mat ing and on Days 1, 5 and 10 of l actation. 2.4.4 Food consumption The weight of food supp lied to each cage, that remaining and an est imate of any spilled was recorded on a weekly basis (fro m the first day of treatme ntuntil pairing). From these records the mean weekly

consumption per animal (g/rat/week) was calculated for each cage. For males in Week 2, to allow acclimat isat io n of the males to the mat ing cages, the males were mo ved to the individual mat ing cages af ter exposure on Day

6 of Week 2. Hence the recorded fo od intake for Week 2 is presented in the appendix over a 6 not 7 day period f or males in Week 2. For tabulated data the food consumpt io n has been adjusted to provide a representative value for the ent i re week. For each F0 female, the weight of food supplied, that remained and an est imate of any spilled was also recorded for the periods Days 0 - 2, 3 -6, 7- 9, 10 -13, 14- 16 and 17 - 19 after mating and Days 1 -4 and 5- 9 of

lactation. From

these records the mean daily consumpt io n (g/rat/day ) was calculated for each animal. 2.4.5 Water consumption Water consumpt io n was recorded by weight (over a 3 day

period on each occasion) on a daily basis fro m Week 2 for each cage of male animals (excluding the pairing period) , and in Week 2 for each cage of female animals, using water bottles fitted with sipper tubes.Huntingdon Life Sciences WAG0015

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### For each F0 female, water consumpt io n was also recorded daily fro m Day 0 after mat ing and fro m Day 1 of lactation, and has been tabulated fo r the periods Days 0 - 2, 3 -5, 6-8, 9-11, 12- 15, 16 - 17 and 18 - 19 after mat ing and Days 1 - 3 and 7 - 9 of lactation. 2.4.6 Oestrous cycles For 15 days before pai

ring, daily vaginal smears were taken from all females, using cotton swabs mo istened wit h saline. The smears were subsequent ly examined to establish the duration and regularit y of the oestrus cy

cle. After pairing with the male, smearing was continued using pipette lavage, unt il evidence of mating was observed. 2.4.7 Mating procedure Fo llo wing a minimum of two weeks of treatment, males and females were paired on a one- to - one basis fro m wi

thin the same treatment group for a period of up to two weeks. Each morning following pai ring, the trays beneath the cages were checked for ejected copulat io n pl

ugs and a vaginal smear was prepared from eac h fe male and examined for the presence of spermatozoa and the stage of the oestrus cy cle. The day

on which evidence of mat ing was found was designated Day 0 of gestation. Once mating occurred, the males and females were separated and smearing was discont inued. The pre - coi

tal int erval was calculated for each female as the time elapsing between init ial pai ring and detection of mating. 2.4.8 Parturition observations and gestation length From Day 20 after mat ing, females were inspected three times daily for evidenc e of parturit io n. The progress and complet io n of parturit io n was monitored, numbers of live and dead offspring were recorded and any difficul ti es observed were noted. The duration of

gestation was calculated as the time elapsing between the detection of m at ing and co mmencement of parturit io n. Half a day was subtracted in cases where parturition commenced overnight. Females were sacrificed on Day 25 af ter mat ing if parturit io n was not complete or had not started. 2.4.9 Records made during littering phase All litters were examined at approximately 24 hours after bi

rth (Day 1 of age) and then daily thereafter. The records maintained were as fo llo ws: Clinical signs: Daily records were maintained for evidence of ill healt h or reaction to treatment; these were on an individual offspring basis or for the litter as a whole, as appropriate. Litter size: Daily records were maintained of mortalit y and consequent changes in litter size from Days 1 -10 of age. Sex ratio: The sex ratio of each litter was recorded on Days 1 , 5 and 10 of age.Huntingdon Life Sciences WAG0015

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### Bodyweight: Individual o ffspring body we ights were recorded on Days 1, 5 and 10 of age. 

## 2.5 Necropsy and histology 2.5.1 Time of necropsy F0 males were killed after Day 10 of lactation of

the majorit y of females. F0 females surviving until the end of the scheduled study period were killed on Day 10 of lactation. Females that failed to produce a viable litter were killed on Day 25 af ter mat ing. Females whose litter died or was sacrificed before Day 10 of lactation were killed on the day the last offspr ing died. Offspring were killed on Day 10 of age , unless they

had shown evidence of ill health. 2.5.2 Method of kill All adul t animals were killed by

carbon dioxide asphyxiat io n. Offspring were killed by intraperi toneal inject io n of sodium pentobarbitone. The sequence in which the animals were killed after complet io n of the study was selected to allow sat is factory

int er - group comparison. 2.5.3 Macroscopic pathology All adul t animals were subject to a detailed necropsy, which involved the following: After a review o f the history of each animal, a full macroscopic examinat io n of the tissues was performed. All external features and orifices were examined visually. After ventral mid - line incisio n, the neck and associated tissues and the thoracic, abdo minal and pelvic cavi ti es and their viscera were exposed and examined in situ. Any abnormal posit io n, morphology or interaction was recorded. External and cut surfaces of the organs and tissues were examined as appropriate. Any abnormalit y in the appearance or size of any organ and tissue was recorded and the required tissue samples preserved in appropriate fixat ive. The requisi

te organs were weighed and external and cut surfaces of the organs and tissues were examined as appropriate. Any abnormali

ty in the appearance o r size of any organ and tissue was recorded and the required tissue samples preserved in appropriate fixat ive. For F0 females, the numbers o f implantation sites in each uterine horn was counted. For females failing to produce a viable litter, the number of uterine implantation si tes was re - checked after staining with ammo nium sulphide (modification of the Salewski staining technique ( Salewski , E, 1964 )). Females whose litter died or was sacrificed before Day 10 of lactatio n were also examined fo r mammary

tissue appearance. For offspring surviving to scheduled terminat io n, a careful external examinat io n was performed f

or gross abnormali

ti es and externally

normal offspring were discarded withoutHuntingdon Life Sciences WAG0015

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### int ernal examinat io n. Externally abnormal offspring were internally examined and any abnormal ti ssues were retained in an appropriate fixat ive. Addit io nally

the f ollo wing procedures were applicable: Premature deaths: Mi ssing offspring and those grossly autolysed or grossly cannibal ised could not be examined. All other offspring dying or sacrificed bef ore Day 10 of age were examined as detailed above. The necropsy also included an assessment for the presence of milk in the stomach, where this was possible. The retained tissues were checked bef ore disposal

of the carcass. 2.5.4 Sperm analysis Im mediately after scheduled sacrifice of each F0 generat io n male, the left vas deferens, epididymis and testis was removed and the epididymis and testis were weighed. The fo llo wing tests were perfo rmed: Sperm motility: A sample of sperm was expressed fro m the vas deferens into pre - warmed (37  C) medium M199, which contained 0.5% w/v bovine serum albumin (BSA Fraction V). A sample for assessment was taken into a 100μm depth cannula by

capillary act io n and where possible at least 200 sperm per animal analysed using the Hamilton Thorne IVOS Computer Assisted Sperm Analyser (CASA) versio n 12.3d. The percentages of motile and progressively motile sperm and sperm mot io n parameters were reported. Sperm morphology: A 200  L aliquot of the sperm/medium mixture (described above) was diluted with 800  L of 10% neutral buffered formalin. After staining wi th nigrosine and eosin an air - dried smear was prepared and examined by light microscopy

f or the assessment of sperm morphology . Where possible at least 200 sperm were assessed for each male. The percentages of normal sperm and abnormal sperm and a summary of observat io ns were reported. Sperm count: The lef

t cauda epididymis of each male was weighed and then froz en. After thawing, 10 mL of a mixture of 0.9% saline, 0.01% merthio lat e and 0.05% Tri ton X - 100 (SMT) was added and the cauda epididymis was ho mo genised for at least one minute. An aliquot of this mixture was added to a pre - prepared IDENT stain tube befor e being assessed for sperm count using CASA. The concentration (Millio n/g) and total number of sperm were reported. Homogenisation - resistant spermatids count: The lef t testis of each male wa s frozen. After thawing, each testis was ho mo genised for at least two minutes in 25 mL of SMT. An aliquot of this mixture was added to a pre - prepared IDENT stain tube before being assessed for homogenisat io n- resistant spermat id count using CASA. The concentration ( M illio n/g) and total number of spermatids were report ed. The codes in the tables and appendices have the following meaning: % Motile - the percentage of cells which are moving at or above the minimum speed as defined in the set up parameters.Huntingdon Life Sciences WAG0015

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### % Progressively motile - the percentage of cells moving with both VAP > progressive minimum VAP and STR > S0. (progressive minimum VAP is a delimiter used in determining whether a motile cell is labelled as rapid or medium. S0 is the threshold straightness.) VAP - the average path velocit y, this is the average velocit y of the smoothed cell path. VSL - the progressive or straight line velocit y, this is the average velocit y measured in a straight line fro m the beginning to the end of the track. VCL - the curvilinear velocit y or track speed, this is the average velocit y mea sured over the actual po intto point track fo llo wed by the cell. ALH - amp litude of lateral head displacement, this is the mean over all cell tracks of twice the maximum displacement between each sperm track and its average path. This corresponds to the average of the sperm track width. BCF - beat cross frequency, this is the frequency wi th which the sperm head crosses the sperm average path. STR - straightness, the average value of the ratio VSL/VAP which measures the departure of the cell path from a stra ig htline. LIN - linearit y, the average value of the ratio VSL/VCL which measures the departure of the cell track from a straight line. Elongation - the average value of the ratio of mino r to maj or axis of all sperm heads. Area - the average size of all sperm heads. Rapid - the fract io n of cells mo ving with VAP > progressive minimum VAP. Medium - the fract io n of cells mo ving with slow VAP cut - off < VAP < progressive minimum VAP. Slow - the f

raction of cells mo ving with VAP< slo w VAP cut - off or VSL < slow VSL cut - off. Static - the f

ract io n of all cells which are not moving at all. 2.5.5 Organ weights The fo llo wing organs, taken from each male, were dissected free of adjacent fat and other contiguous ti ssue and the weights recorded: Epididymides (L&R) Testes (L&R) Pi tui tary Prostrate Seminal vesicles L&R Bilateral organs weighed individually 

### Organ weights were also adjusted for terminal bodyweight, using the weight recorded before necropsy .Huntingdon Life Sciences WAG0015

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### 2.5.6 Fixation Testes and epididymides were init ially fixed in modi fied Davidson's fluid prior to transfer to 10% neutral buffered formalin . Sam ples (or the whole) of

the other tissues listed below from all anima ls were preserved in 10% neutral buffered formalin: For adul t animals: Abnormali

ti es Prostate Epididymides+ Seminal v esicles # Mammary area - caudal†# Testes+ Ovaries Uterus wi th cervix and oviducts # Pi tui tary‡# Vagina# # Not examined histologically † Females with total litter loss only ‡ For males with suspect fertility only + Only right fixed and subject to histo logy and pathology; left organ used for sperm analysis 

### Sam ples o f any abnormal t is sues were also retained and processed for examinat io n. In those cases where a lesio n was not clearly delineated, cont iguous tissue was fixed with the grossly affected region and sect io ned as appropriate. Sam ples o f the mammary area (from females wit h total litter loss only), pi tui tary (f rom males of suspect fertilit y only), uterus with cervix and oviducts and vagina were not examined histologically, but are retained again st any f uture requi

rement for microscopic examinat io n. 2.5.7 Histology For those animals specified in the Patho lo gy sect io n, the relevant tissues were subject to histological processing. Tissue samples were dehydrated, embedded in paraffin wax, sect io ned at approximately f our to five micron thickness and stained with haematoxylin and eosin. The testes were stained using a standard periodic acid/Schiff (PAS) method. Those tissues subject to histological processing included the fo llo wing regions: Epididymides - caput, corpus and cauda Ovaries - qualitat ive evaluat io n of one sect io n from each ovary For bilateral organs, sections of both organs were prepared. A single section was prepared fro m each of the remaining tissues required for microscopic pathology.Huntingdon Life Sciences WAG0015

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## 2.6 Path ology 2.6.1 Light microscopy Microscopic examination was performed as follows: All t issues preserved for examinat io n (as specified above) were examined for all animals o f Groups 1 (Control) and 4 (2900 ppm ) sacrificed on com

plet io n of the scheduled treatment perio d and for all animals killed or dying during the study. Tissues reported at macroscopic examinat io n as being grossly abnormal were examined for all animal s in line wit h current practice ( incisors or skin showing hair lo ss is not examined). The fo llo wing tissues, which were considered to exhibit a reaction to treatment at the high dose, were examined for all animals : Ovaries and Prostate. Findings were either reported as "present" or assigned a severit y grade. In the latter case one of the fo llo wing five grades was used - minimal, slight, moderate, marked or severe. A reviewing pathologist undertook a peer review o f the microscopic findings. For the assessment of the ovaries, one mid- line section of each ovary was examined for the presence of primordial fo llicles, growing fo llic les and corpora lutea. For the assessment of the right testis, a detailed qualitat ive examinat io n was made, taking into account the tubular stages of the spermatogenic cycle. The examinat io n was conducted in order to ident ify tr eatment related effects such as missing germ cell layers or ty pes, retained spermat id s, m

ul tinucleate or apoptotic germ cells and sloughing of spermatogenic cells into the l umen. Any cell- or stage - specificit y of testicular findings was noted. For the ass essment of the epididymides, leukocy te infil tration, change in prevalence in cell ty pes, aberrant cell t ypes and phagocytosis of sperm was examined. 

## 2.7 Computer systems The computer systems wi th version numbers that were used on this study to acquir e and qu ant ify data include: Hamilton Thorne IVOS Computer Assisted Sperm Analyser (CASA) (version 12.3d) used for Sperm analysis. Liberate Release 2 in- house system used for reporting clinical signs, dosing signs, body weights, food consumpt io n, water consumpt ion , organ weights, macropathology

and histopathology, statist ics. Quasar (versio n 1.3) and SAS (versio n 9.1.3) in- house statist ical analysis packages used for body

weight, food consumption and organ weight statist ic s. Sam ple Registry

System (versio n 1.0) used fo r Formulat io n and Inhalat io n Analysis data. SAS (versio n 9.1.3) in house statist ical analysis package used for sex ratio statist ics.Huntingdon Life Sciences WAG0015

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### StarTox (version 3.2) in- house statist ical analysis package used for litter size and survival indices statist ic s. StatX act 3 statist ical analysis package used for mating performance and fertilit y and oestrous cycles and pre - coi tal int erval statist ic s. Waters Empower 2, Build 2154, Feature Release 4, Service Pack D and used for Form ulat io n and Inhalat io n Analysis . Xybio n Path

/Tox System (versio n 4.2.2) used for clinical signs, dosing signs, body weights, food consumpt io n, water consumption, organ weights, macropathology

and histopathology . Xybio n Pri st ima (version 6. 3. 2) used for Pharmacy test substance management. 

## 2.8 Data trea tment This report contains serial observations pertaining to all weeks of treatment completed, together with data collected during the necropsy period. Summary statistics (e.g. means and standard deviatio ns) presented in this report were calculated from

co mputer - stored individual raw data. The summary statist ic s and the individual data were stored in the computer to a certain number of decimal places, different fo r each parameter. For presentation purposes, however, they were usually

rounded to fewer plac es. It is, therefore, not generally possible to reproduce the presented means and standard deviat io ns exactly using the presented individual data. The codes in the appendic es have the fo llo wing meaning: T Scheduled kill; fertili ty confirmed W Killed for reasons of animal welfare F Found dead FTM Failed to mate NP Not pregnant; no implantation sites visible TR Total

resorption; implantation si tes visible TLL Total

litter loss U Unilateral pregnancy Wit hin the context of this study i t was not 100% possible to confirm animals that were pregnant but where no pups were seen in the animal’s cage, were total resorptions (litter loss in utero) or if any

pups had been born but had been shortly afterwards eaten by the dam. These animals have been coded as TLL with the above proviso/caution. Throughout the report the fo llo wing abbreviat io ns are used: M Male F Female N/n Number contribut ing to the mean (normally the number animals/litters) SD Standard deviat io nHuntingdon Life Sciences WAG0015

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### 2.8.1 Signs Signs are considered in two parts: observat io ns in associat io n wi th treatment, classified as “dosing signs” and extended changes in condit io n, classified as “clinical signs”. Signs considered to indicate a reaction to treatment are presented for each animal that shows signs, providing detail

of ty pe of sign, day

of occurrence and information on the duration of the sign if applicable. “Dosing signs ” have been presented in 2 ways, one to show the daily occurrence relative to the timing of exposure, and one to show progression/presence across the differe ntphases of the study . Dosing signs recorded during the 6 hour exposure were recorded on a group basis; hence in this report all animals in a group will exhibit the same signs. Signs associated with dosing recorded prior or post exposure are recorded on an individual animal basis. The overall group frequency o f dosing signs and clinical signs were summarised in an incidence table. 2.8.2 Bodyweights Individual values were reported for all days recorded. Group mean weight changes were calculated from

the weig htchanges of individual animals surviving the specified period. Body weight was plotted graphically wi th respect to the f

ir st day of the relevant period . 2.8.3 Food consumption During the pre - pair ing periods of the study where animals were housed 5 per cage, val ues are presented for the average amount of food consumed per rat in each cage in each experimental week (g/rat/week). Values presented allow for any animal that died or was killed during the week. For males in Week 2, to allow acclimat isat ion of the males to the mat ing cages, the males were moved to the individual mat ing cages after exposure on Day 6 of Week 2. Hence the recorded food intake for Week 2 is presented in the appendix over a 6 not 7 day period f or males in Week 2. For tabulated data the f ood consum

pt io n has been adjusted to provide a representative value for the ent ir e week. During the gestation and lactation phases, where females were individually housed, group mean values and SD were calculated fro m individual values for Days 0 - 2, 3 -6, 7-9, 10- 13, 14- 16 and 17 - 19 after mat ing and Days 1 - 4 and 5 - 9 of lactation based on the numbers of females co mp let ing each phase. Mean overall values for food consumpt io n before pairing were calculated fro m the weekly group mean values presented. 2.8.4 Water consumption Water consumpt io n was calculated fro m me asurements of init ial and final weights (g) of the water bottle and contents for each cage (it was assumed that 1 mL of water weighed 1 g).Huntingdon Life Sciences WAG0015

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### 2.8.5 Oestrous cycles The percentage females showing the fo llo wing cl assifications of oestrous cy

cles before pai ring are presented: Regular: All observed cycles of 4 or 5 days Irregular: Atleast one cycle of 2, 3 or 6 to 10 days Acyclic: Atleast 10 days without oestrus (beginning before pairing). 2.8.6 Pre - coital interval Individual

int ervals were tabulated for the time elapsing between init ial pairing and mat ing. Percentage of females wit h pre - coi tal int ervals calculated for durations of 1 - 4, 5 -8, 9- 12 and 13- 14 days of pairing. 2.8.7 Mating performance and fertility Individual data was tabulated. Group val ues were calculated for males and females separately f or the following: Percentage mating = Number animals mating x 100 Animals paired Conception rate (%) = Number animals achieving pregnancy x 100 Animals mated F ertility index (%) = Number animals achieving pregnancy x 100 Animals pairing 

### 2.8.8 Gestation length Gestation length was calculated as the number of gestation days up to and including the day on which offspring were first observed, with Day 1 = day of mating for calculation purposes. Where parturit io n had started overnight, this value was adjusted by subtracting half of one day . Gestation index was calculated for each group as:

Gestation index (%) = Number of live litters born x 100 Number pregnant 

### 2.8.9 Litter size Individual litter values were tabulated for the number of implantation sites, total at Day 1 (live and dead) and total live on Days 1 , 5 and 10 of age. Group mean litter size and SD were calculated from

the individual litter values.Huntingdon Life Sciences WAG0015

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### 2.8.10 Surviva l indices The fo llo wing were calculated for each litter:

Post - implantatio

n survival index (%) = Total number of offspring born x 100

Total number of uterine implantation sites 

### Post- implantation survival index was expressed as 100% where the number of offspring exceeded the number of implantation sites recorded.

Live birth index (%) = Number of live offspring on Day 1 after littering x 100 Total number of offspring born Viability index (%) = Number of live o ffspring on Day 10 after littering x 100 Number live offspring on Day 1 after littering 

### Group mean values were calculated from individual litter values. 2.8.11 Sex ratio The percentage of male o ffspring in each litter was calculated at Day 1, and for live o ffspring on Days 1 , 5 and 10 of

age. Percentage males = Number of males in litter x 100

Total number of offspring in litter 

### Group mean values were calculated from individual litter values. 2.8.12 Sperm analysis Individual values tabulated for the fo llo wing: Motilit y (vas deferens) The percent age of motile and progressively moti le sperm and a detailed assessment of motilit y were reported. Morphology The number and percentages of normal and abnormal sperm were reported. A summary of the ty pes of abnormali ti es seen was made because there was evidence of an effect on the percentage normal sperm. Count (Testis and Cauda epididymis) The sperm concentration (Millio n/g) and total number were reported. Group mean values were calculated from individual values. 2.8.13 Organ weights The abbreviat io ns used in the Appendix have the following meanings: Lt Left Rt RightHuntingdon Life Sciences WAG0015

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### 2.8.14 Pathology The abbreviat io ns used in the Appendix have the following meanings: Fac. Con. Factors contribut ing Lt. Left Rt. Right Tissues which could not be examined are specified in the Append ix. The absence of a comment for a tissue scheduled for examinat io n therefore indicates that the tissue was considered to be normal. In all tabular presentations o f data the tissues specified in the protocol f or hi stopathological examinat io n precede other tissues. For offspring, findings were presented on an individual basis. 2.8.15 Statistical analysis All statist ical analyses were carried out separately for males and females. Data relat ing to food consumpt io n during the pre - pair ing period were analysed on a cage basis. For all other parameters, the analyses were carried out using the individual animal as the basic experimental unit. The fo llo wing data ty

pes were analyse d at each timepoint separately: Body weight, using absol

ute weights and gains over approp riate study periods Food consumpt io n, over appropriate study periods Organ weights, both abso lut e and adjusted for terminal body weight Li tter data and survival indices Mating performance and fert ilit y The fo llo wing sequence of statist ical tests was used f or body weight, f ood consum

pt io n, organ weights , litter size and survival indices data : A parametric analysis was performed if Bartlett's test for variance homogeneit y ( Bartlett 1937 ) was not significant at the 1% level. F or pre - treatment data, analysis of variance was used to test for any group differences. Where this was significant ( p <0.05) inter group comparisons using t- tests, wi th the error mean square fro m the one- way analysis o f variance, were made. For all other analyses the F 1 

### approximate test was applied. This test is designed to detect significant departure from mo notonici

ty of means when the main test for the comparison of the means is a parametric monotonic trend test, such as Williams’ test ( Williams 1971, 1972 ) . The test statistic compares the mean square, NMS, for the deviat io ns of the observed means from the maximum likelihood means, calculated under a constraint of mo notonici

ty wi th the usual error mean square, EMS. The null hypothesis is that the true means are monotonically ordered. The test statist ic is F 1 

### = NMS/EMS which can be co mpared with standard tables of the F - distribution wi

th 1 and E MS degrees of f reedo m. If the F 1 

### approximate test for monotonicit y of dose - response was not significant at the 1% level , Williams' test for a monotonic trend was applied. If theHuntingdon Life Sciences WAG0015

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### F 1 

### approximate test was significant, suggesting that the dose - response was not mo notone, Dunnett's test ( Dunnett 1955, 1 964 ) was performed instead. A non- parametric analysis was performed if Bartlett's test was still significant at the 1% level fo llo wing both logari thmic and square - root transformat io ns. For pre - treatment data, Kruskal- Wallis’ test (Kruskal and Wallis 195 2, 1953) was used to test for any

group differences. Where this was significant ( p <0.05) inter group com parisons using Wilcoxon rank sum tests (Wilcoxon 1945) were made. For all other analyses the H 1 

### approximate test, the non- parametric equivalent of the F 1 

### test described above, was applied. This test is designed to be used when the main test fo r com parison of the means is a non- parametric monotonic trend test, such as Shirley's test ( Shirley 1977 ) . The test statist ic co mpares the non- mo notonici ty sums of squares, NRSS, for the deviat io ns of the observed mean ranks from the maximum likelihood mean ranks wit h the non- parametric equivalent of the error sums of squares, ERSS = N(N+1)/12. The test statistic is H 1 

### = NRSS/ERSS which can be com pared to standard tables of the χ 2



### - distribut io n wi

th 1 degree of freedo m. If the H 1 

### approximate test for monotonicit y of dose - response was not significant at the 1% level, Shirley's test for a monotonic trend was applied. If the H 1 

### approximate test was significant, suggesting that the dose - response was not monotone, Steel's test ( Steel 1959 ) was performed instead. For litter data , if 75% of the data (across all groups) were the same value, for example c, Fisher’s Exact tests ( Fi sher 1973 ) were performed. Treatment groups were compared using pai rwise com

parisons of each dose group against the control both for i) values <c versus values ≥c, and for ii) values ≤c versus values >c, as applicable. Sex ratio were analysed by generalised mixed line ar model wi th bino mial errors, a logit link funct io n and litter as a random effect ( Lipsitz et al 1991 ) . Each treated group was compared to con trol using a Wald chi- square test. For sex ratio , the numerator was Number o f ma les, the deno minator was Number of live fetuses. For gestation length, sperm counts and copulation plugs an exact two - tailed Linear - by- linear test ( Cyt el

1995 ), wi

th equally spaced scores, was applied to all groups. If the test was statist ically significant (

p <0.05), the highest dose group was excluded and the test re - applied. This ‘step - down’ process was repeated until the test was no longer statist ically significant ( p ≥0.05). For the number of females wit h one or more ‘abnormal’ oestrous cycle length of ‘3 or less’ or ‘5 or more’ days an exact one - tailed (upper - tail) Cochran- Armit age test ( Cyt el 1995 ) was applied to all groups. If the test was statist ic ally significant ( p <0.05), the highest dose group was excluded and the test re - applied. For the incidence of cycle lengths an exact one - tailed (upper - tail) Linear -by- linear test ( Cy tel 1995 ) was applied to all groups, using scores appropriate to the severit y of the observat io n assuming 4 day

cycles to be normal . The categories were scored as follows: a 4 day cy cle was scored as 4 and a 5 day cycle was scored as 5 and abnormal ( 2, 3, 6 to 10 or 11 to 20 days) cycles were scored as 6 . If the test was statist ically significant ( p <0.05), the highest dose group was excluded and the test re - applied. This ‘step -down’ process was repeated until the test was no longer statist ically significant ( p ≥0.05).Huntingdon Life Sciences WAG0015

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### For pre - coi

tal int ervals an exact one - tailed (upper - tail) Linear -by- linear test ( Cyt el 1995 ) was applied to all groups, using the fo llo wing scores: a 1 - 4 day int erval was scored as 1, a 5 - 8 day int ervals was scored as 2, a 9 - 12 day int erval was scored as 3 and a 13 - 14 day int erval was scored as 4. If the test was statist ically significant ( p <0.05), the highest dose group was excluded and the test re - applied. This ‘step - down’ process was repeated until the test was no lo nger statist ically significant ( p ≥0.05). For number mat ing, number conceiving, number fertile and number of live litters born an exact one - tailed (lower - tail) Cochran- Armit age test ( Cyt el 1995 ) was applied to all groups. If the test was statistically signif icant ( p <0.05), the highest dose group was excluded and the test re - applied. This ‘step down’ process was repeated until the test was no longer statist ically significant (

p ≥0.05). If the exact version o f the Linear -by- linear test could not be calculated ( due to the size of the table containing the data), then the asymptotic versio n was used instead. Key to Tables Significant differences between Control and treated groups were expressed at the 5% ( p <0.05) or 1% (

p <0.01) level. The key to the annotation use d on the tables that contain statist ical results is given below. l Data were log transformed for the statist ical analysis. Ca Treated groups compared to Control using Cochran- Armit age test Du Treated groups co mpared to Control using Dunnett’s test. Lt Tre at ed groups compared to Control using Linear -by- linear associat io n test Sh Treated groups co mpared to Control using Shirley’s test. Wa Treated groups compared to Control using Wald’s test Wi Treated groups co mpared to Control using Williams’ test. Codes p laced above adjusted means indicate that the co mpar isons were based on adjusted means. * p <0.05 ** p <0.01 

## 2.9 Quality assurance and archiving procedures 2.9.1 Quality assurance Details o f the Qualit y As surance inspections and audits are presented on the Qualit y Assurance Statement. 2.9.2 Archives Fo llo wing com

plet io n of this study

all raw data, specimens and samples, except those generated or used during any Sponsor’s or supplier’s analysis, were stored in the archives o f Hunt ingdon Life Sciences. Types of sample and spe cimen which are unsuitable, by reason ofHuntingdon Life Sciences WAG0015

34 

### instabilit y, f

or long term

retention and archiving may b e disposed of after the periods stated in Hunt ingdon Life Sciences Standard Operating Procedures. A copy of the final report and all

Quali ty A ssurance inspect ion records will be retained indefinit ely. All other appropriate specimens and records will be retained fo r a minimum period of one year fro m the date of issue of the final report. At the end of this retention period the Sponsor will be contacted and advice sought on future archiving requirements. Under no circumstances will any item be discarded without the Sponsor’s knowledge. 

## 2.10 Deviations from protocol The fo llo wing deviat io ns from protocol occurred: In Week 6 for females, the water consumption residues were not recorded in error and there was thus no data for Week/Day 6.4 to 6.7. As such this was repeated over Week/Day

7.1 to 7.3. There was therefore two sets of water consumpt io n, each recorded over 3 days, in Week 7. The animals were supplied at age 49- 56 days and weight range 250 - 300g males and 200- 230g females, not at 65 days and 310 - 350g males, 227 - 254g females as required by the protocol. As such the animals were approximately 8 - 9 weeks of age at the start of treatment, not 10 weeks. The pi tui t ary, prostates and seminal vesicles (with coagulat ing gland) from all males were weighed, not j ust those of suspect fertilit y. The sex ratio of litters was also calculated and reported on Day 5 post partum, not just Day 1 and Day

10. Water consumpt io n wa s presented daily f or individual

values. Water consumpt io n was tabulated on a weekly b asis for males, and for females prior to pairing, and tabulated for females during gestation and lactation both on a daily basis and over the period specified in the pro tocol . These deviat io ns w ere considered to have not affected the integrit y of the study .Huntingdon Life Sciences WAG0015

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# 3. Results 3.1 Chamber atmosphere conditions Annex 2 The achieved chamber concentration data for 2 - bromo - 3,3,3 - trifluoropropene after approximately

8 weeks of treatment are summarised as fo llo ws: Group Treatment Chamber concentration (ppm) Target Achieved 1 Control 0 - 2 2 - br omo - 3,3,3 -

trifluoropropene 200 198 3 2 - br omo - 3,3,3 -

trifluoropropene 500 505 4 2 - br omo - 3,3,3 -

trifluoropropene 3000 2900 

### The achieved aerosol

concentrations were 99, 101 and 9 7 % of the target concentration for Groups 2, 3 and 4, respectively. Adjust me nt s were made to the exposure systems as appropriate to ensure that the achieved aerosol concentrations were close to target. 

## 3.2 F0 maternal responses 3.2.1 M ortality and animal fates Table 1 At505 ppm, 5 f

emales and litters were sacrificed during lactation. In these cases the litter had a high number of dead pups, and the offspring tended to show signs of poor condit io n, being co ld with reduced act ivit y and were killed wit h the dams. In one case, all pups that were born were found dead at the completion o f parturit io n check (total litter loss pre - Day 1). At2 90 0 ppm, 2 f

emales were sacrificed on Day 24 af ter mating. Female 276 had partially closed eyes, hunched posture and piloerect io n, and female 279 had piloerect io n and perigeni tal staining and was noted to be attempting parturit io n . Both females were pregnant. The single dam (f

emale 273) wi

th a live litter born at 2 90 0 ppm was killed for reasons of animal welfare fo llo wing total litter loss. 3.2.2 Signs Table 2-3 and Appendi ces 1-4 Appendi ces 1 and 2 show signs noted at the physical examinat io n. Appendix 3 shows signs during and after exposure by phase of the study; the signs noted during exposure apply to all animals in the group being exposed . Appendix 4 shows , f or f emales, the incidence of individual signs associated with dosing as the study progressed prior to mating and through gestation and lactation.Huntingdon Life Sciences WAG0015

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### Pale teeth were noted in all males and 9 /10 fe males at 2 90 0 ppm (first noted from Week 5 in some and present from Week 7 in all animals) and in 3/10 males and 3/10 females at 505 ppm (only noted in Week 8, the final week of the study ). Hai rloss was noted at a higher incidence than controls in mal es at 2900 ppm . Occasio nally, in some f

emales at 2 90 0 ppm, piloerection, hunched posture, and underact ivit y were noted at the physical examinat io n, generally performed weekly. During the 6 hour exposures, the fo llo wing signs were noted: At2 900 ppm under activit y, unresponsiveness, pilo erection and eyelids partially closed were noted throughout. In males these signs were not apparent after completion o f exposures. In females so me o f the above signs and also hunched posture were noted following exposure, occasionally

in Week 2 (hunched posture only) and generally throughout gestation, but these were not present at the check for signs at the end of the working day (up to 2 hours later). At505 ppm, underact ivit y and piloerection were noted throughout, and unresponsiveness and eyelids partially

clos ed were noted on fewer occasions. In males the signs were not noted fo llo wing exposure, but in females during gestation and lactation, underact ivit y and piloerect io n, and in females during gestation only, eyelids p artially clos ed were noted after exposure but not at the end of the working day . At19 8 ppm underactivit y, unresponsiveness, piloerection and part ia lly clos ed eyelids were noted at a much reduced incidence than that seen at higher levels; only underact ivit y and piloerect io n were noted immediately after exposure during gestation only. 3.2.3 Bodyweight Figure s 1-5 , Table s 4-7 and Appendi ces 5-8 Mean body

weight gain of males over the treatment period was lower than the controls in all groups gi ven the test article ( gain Day 1 - 50: 77, 63 and 47% of control at 19 8 , 505 and 2900 ppm respectively). The effect was apparent from Week 1. Mean body

weight gain of females over the 2 week period prior to pairing was lower than the controls at 2 90 0 ppm

(gain Day

1 - 15: 19% of con

trol). There were no effects of treatment on body weight gain of females over the 2 week period prior to pairing at 198 or 505 ppm. Group mean body

weight on Day

0 of gestation f

or females at 2 90 0 ppm

was lower than the controls, reflect ing the difference e stablished prior to pai ring. Mean body

weight gain during gestation was lower than the controls in females at 505 and 2 90 0 ppm (gain Day

0 - 20: 83 and 43% of control respect ively). At19 8 ppm, mean body weight gain was marginally lower than the controls dur ing Days 0 - 20 (92% of control). During lactation, at 505 ppm slight mean body

weight loss occurred once treatment was reintroduced on Day

5 of lactation (2g loss com

pared to 20g gain in the control group), and as such overall bodyweight gain during lactatio n was lower than the controls (13% of control). At19 8 ppm bodyweight gain during Days 1 - 10 of lactation was 59% that of the control. No litters survived at 2 900 ppm and hence no adult female body weights were recorded.Huntingdon Life Sciences WAG0015

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### 3.2.4 Food consumption Table s 8-10 and App endi ces 9-11 Mean food consumpt io n of males and females was lower than the controls in all treated groups over the 2 weeks prior to pairing, with the effect most marked in Week 1. Overall fo od consumption for the 2 week period prior to pairing was 88, 76 and 76% of controls for males and 92, 82 and 70% of controls for females at 19 8 , 505 and 2 90 0 ppm respect ively. During the gestation phase, food consumpt io n of treated pregnant females remained low and at a similar magnitude to that noted prior to pairing fo r treated females co mpared with controls (Day

0 - 19: 90, 8 9 and 78% of control at 19 8 , 505 and 2 90 0 ppm ). During the lactation phase, food consumpt io n was lower than controls for females at 19 8 and 505 ppm (73 and 60% of control respect ively). No litters survived at 2900 ppm and hence no fo od consumption data was generated for lactating females. 3.2.5 Water consumption Table s 11-13 and Appendi ces 12-14 Water consumpt io n was recorded for ma les and females from Week 2 of the treatment phase onwards following a vi sual increase in consumption noted for females in Week 1. Measured water consumption of f emales in Week 2 was 17, 5 9 and 72% higher than the controls at 19 8 , 505 and 2 90 0 ppm respect ively. During the gestation phase, water consumption was slight ly higher than the controls for all groups of treated females from Day 3 (up to 23% higher than the controls in females at 505 ppm ) , al though the magnitude of the difference was less marked than that recorded prior to pai ring. During the lactation phase, water consum pt io n , once treatment was reintroduced on Day 5, was slight ly lo wer f

or f emales at 19 8 and 505 ppm compared with control . There were no effects on water consumpt io n of males in Weeks 2, 5, 6 or 7 (water consumption was not recorded during the 2 week pair ing period). 3.2.6 Oestrous cycle length Table 14 and Appendix 15 There was an effect on oestrous cycles in females. It is expected that control and unaffected animals have a preponderance of 4 or 5 day oestrous cycles , considered regular . All control females and females at 198 ppm had only 4 or 5 day

cycles. At2900 ppm, only

1 female had regular cy

cle lengths, al

though the cycle lengths recorded fo r this f emale were all 5 days in durat io n. Three females also had irregular cycles (at least one cy cle of 2 or 3 days, or 6 - 10 days), one female had a period of extended oestrus, and the remaining 5 females were considered to also have, or only be, acyclic (at least 10 days wi thout oestrus). Assessment of cycle lengths showed, in females at 2900 ppm, a statist icallyHuntingdon Life Sciences WAG0015

38 

### significant incidence of females wit h lo nger cycle lengths, and a statistically significant incidence of longer specific cycle lengths, with 46% of the total recorded cy cle lengths being 6 to 10 days, and 18% of the recorded cycle lengths being 11 - 20 days, com pared wi th no similarly longer cycle lengths being observed in the control group. Only 33% of the total recorded cy

cle lengths at 2900 ppm were regular (4 or 5 days) compared to all o f those in the control group. At505 ppm, only 5/10 females had re gular cycle lengths, although in 4 of these 5 animals the cy cle lengths were all 5 days. Four females had one or more irregular cycles and one female was acyclic (wi

th a period of

12 days wi

thout oestrous). Assessment of the cycle lengths showed a statist ically significant incidence of females with longer cycle lengths, and a statist ically significant incidence of lo nger specific cy cle lengths, wi

th 21% of recorded cycle lengths being 6 or more days long. Al though 79% of the recorded cycle lengths were considered regular (compared to 100% in the control group), 61% were of 5 days duration com pared to only

18% of those in the control group. At198 ppm, all

f emales had regular cy

cle lengths (4 or 5 days) , and there was thus considered to be no adverse effect . There was, however, a statistically significant shift towards the lo nger cycle length with 70% of all cycles being 5 days duration co mpared with only 18% of the control

cycles. 3.2.7 Pre - coital interval , mating performance and fertility Table s 15-18 and Appe ndix 15 At2 90 0 ppm, only

7 of the pai rings had evidence of mat ing and of these only 4 occurred in the first 4 days after pairing (the time considered to generally reflect a mat ing at the first oestrous possibilit y) . However, review of the time of success fu l mating showed that 6 of the 7 females mated at the first oestrus recorded after paring with only one mating at the second oestrous cycle. There w as a statist ically significant trend towards lower numbers of copulat io n pl

ugs at mat ing. Copulation plug s give an indicat io n of the number of times that mat ing occurred . Only 29% of the mat ings had 4 - 6 recorded copulat io n pl ugs com pared to 80% of the controls; the remaining 71% of mat ings had 2 or 3 copulat io n pl ugs. Sperm count estimated at mating showed a slight ly lo wer estimate of the presence of sperm in the vaginal smear taken on the day

of mating, wi

th the maj ori

ty of smears having continuous few sperm or many scattered sperm compared to the solid masses of sperm seen more frequent ly in the control group. Of the 7 pairs o f ma les and females showing evidence of mat ing, 6 pairings produced a pregnancy. At505 ppm

all 10 pai rings showed evidence of mating but 3 of these were in days 5 - 8 af ter pai ring, not as would be usually expected in the first 4 days af ter pai ring. A review of the time of successful mat ing showed that all but one females mated successfully at the first oestrous possibilit y. There was a statistically significant trend towards lower number s of copulat io n pl

ugs at mat ing. Only 20% of the matings had 4 - 6 recorded copulat io n plugs com pared to 80% of the controls; the remaining 80% of mat ings had 1, 2 or 3 copulat io n pl ugs. Sperm

count estimated at mat ing showed a trend towards slight ly lo wer estimate s of the presence o f sperm

in the vaginal

smear taken on the day

of mat ing, with the majorit y of smears having cont inuous few sperm or many scattered sperm compared to solid masses of sperm as seen more f

requent ly in the control group. Four of the mat ings had no sperm presentHuntingdon Life Sciences WAG0015

39 

### in the vaginal smear; in the absence of this occurrence at 2900 ppm, this specific finding is of uncertain significance. All 10 pairings were successful in producing a pregnancy . At19 8 ppm and in the control group, the majorit y of animals showed evidence of mating (pre -c oi tal int erval) in the first 4 days after pairing (i.e. at the first mat ing opportunit y) . With the except io n of a trend towards a lo wer sperm count category in the vaginal smear on the day of successful mat ing, there were no effects on the number of copulat io n pl ugs, or mat ing performance. 3.2.8 Gestation length and gestation index Table 19 and Appendix 15 The duration of

gestation in females fro m all treated groups was longer than the controls. In the control group, the females showed the ty pical pattern of a majo ri

ty of gestation lengths of 22 days. Females at 19 8 ppm had gestation lengths of 23 days, and those at 505 ppm had gestation lengths o f 23, 24 or 25 days. After 25 days o f gestation only one f emale at 2900 ppm had co mp leted parturition. The gestation index was reduced at 2 90 0 ppm (only 17%) reflect ing the single litter born fro m those mat ing in this group . 3.2.9 Sperm analysis Tables 2 5-27 and Appendices 23 -25 No adverse effects on percent motile sperm or testicular spermat id numbers were seen after treatme ntwi

th 2 - bro mo -3,3,3- trifluoropropene at exposure levels up to 290 0 ppm. Slight but statist ically significant reductions in percent progressively motile sperm, velocit ies and percent medium sperm were observed after treatment at 290 0 ppm. In addi tion a statist ically significant increase in B eat Cross Frequency was seen at this level . Significant reductions in velocit ies were also apparent at the lower exposure levels of 198 and 50 5 ppm . A slight but statist ic ally significant reduction in number of sperm in the cauda epididymis was apparent after treatment at 290 0 ppm. This correlated with a lower cauda epididymal weight; the sperm

/g was however unaffected. Slight reductions in percent normal sperm were observed in all treated groups, these changes wer e statist ic ally significant at 50 5 and 2900 ppm. The corresponding increase in percent total abnormal sperm

was significant at all exposure levels. Observed abnormalit ies were primarily breakages (including decapitate sperm) and abnormal head shape. The increased B eat Cross Frequency suggests there was no reduction in vigour of the sperm but that their forward movement was affected, whilst the breakages suggest sperm fragilit y was increased.Huntingdon Life Sciences WAG0015

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### 3.2.10 Organ weights Table 28 and Appendix 26 At2 90 0 ppm, several

organ weights were lower than the controls, notably prostate (52% of control), seminal vesicles (71% of control), epididymides ( 87 % of control) and pi

tui tary (77% of control). At505 ppm, prostate (74% of control), seminal vesicles (70% of control) and pituitary (85% of control) weights were reduced. At19 8 ppm, prostate (77% of control), seminal vesicles (83% of control) and pituitary (85% of control) weights were reduced. 3.2.11 Macropathology Table 29 , Appendix 27 and Annex 3 3.2.12 Animals killed after scheduled treatme nt period The macroscopic examinat io ns performed in the F0 animals revealed intergroup differences in the prostate, spleen, incisor teeth and skin. Prostate A reduction in size of the prostate was seen in all males exposed to 2900 ppm, in the majority of males exposed to 505 ppm and in one male exposed to 198 ppm. Summary of findings in the prostate of F0 animals after scheduled treatment period Group/sex 1M 2M 3M 4M Exposure Level (ppm) 0 198 505 2900 Small 0 1 7 10 Number of animals examin ed 10 10 10 10 

### Spleen Capsular thickening was seen in the spleen of the majorit y of males exposed to 2900, 505 or 198 ppm and in occasio nal females exposed to 2900 or 505 ppm. Capsular adhesions were seen in occasio nal treated males across all gro ups, in a few females exposed to 2900 ppm and one female exposed to 505 ppm. Summary of findings in the spleen of F0 animals after scheduled treatment period Group/sex 1M 2M 3M 4M 1F 2F 3F 4F Exposure Level (ppm) 0 198 505 2900 0 198 505 2900 Capsule thickened 0 8 6 9 0 0 2 3 Adhesions 0 2 3 4 0 0 1 4 Number of animals examined 10 10 10 10 10 10 9 8Huntingdon Life Sciences WAG0015

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### Teeth Pale incisor teeth were seen in all male and female animals exposed to 2900 ppm, in the majorit y of

males and three fe males exposed to 505 ppm and in one control male. Summary of findings in the teeth of F0 animals after scheduled treatment period Group/sex 1M 2M 3M 4M 1F 2F 3F 4F Exposure Level (ppm) 0 198 505 2900 0 198 505 2900 Incisor(s) pale 1 0 6 10 0 0 3 8 Number of animals examined 10 10 10 10 10 10 9 8 

### Skin An increased incidence of hair loss was noted in occasio nal males exposed to 2900ppm. Summary of findings in the skin of F0 animals after scheduled treatment period Group/ sex 1M 2M 3M 4M 1F 2F 3F 4F Exposure Level (ppm) 0 198 505 2900 0 198 505 2900 Hair loss 0 1 2 4 2 2 0 0 Number of animals examined 10 10 10 10 10 10 9 8 

### The nature and incidence of all other findings were consistent with the commo n background of macroscopic changes seen at these laboratories. 3.2.13 Histopathology Table 30 , Appendix 27 and Annex 3 3.2.14 Animals killed after scheduled treatment period Treatment related findings Changes related to treatment with 2 - bromo- 3,3,3 - trifluorop ropene were seen in the F0 ovaries. Ovary A slight reduction in the size of corpora lutea was seen in the ovaries of 4 females exposed to 2900 ppm and in a single female exposed to 198 ppm.Huntingdon Life Sciences WAG0015

42 Summary of findings in the ovary of F0 animals after scheduled tre atment period Group/sex 1F 2F 3F 4F Exposure Level (ppm) 0 198 505 2900 Reduced Size of Corpora Lutea Slight 0 1 0 4 Total 0 1 0 4 Number of tissues examined 10 10 9 8 

### Findings of an uncertain relationship to treatment Findings of

an uncertain relat io nship to treatment were identified in the spleen. This tissue was examined microscopically only when macroscopic abnormali ti es had been observed at necropsy . No spleen fro m control animals was examined. Spleen Minimal to moderate capsular/subcapsular inflammat io n and/or capsular thickening and/or adhesio ns/infla mmat io n/ fibrosis were observed in the majorit y of treated males exposed to 198, 505 or 2900 ppm and in occasional females exposed to 505 or 2900 ppm. Summary of findings of an uncertain relationship to treatment in the spleen of F0 animals after scheduled treatment period Group/sex 1M 2M 3M 4M 1F 2F 3F 4F Exposure Level (ppm) 0 198 505 2900 0 198 505 2900 Capsular/Subcapsular Inflammation Minimal 0 5 3 3 0 0 1 3 Slight 0 3 2 2 0 0 0 1 Moderate 0 0 1 2 0 0 0 0 Total 0 8 6 7 0 0 1 4 Capsular Thickening Minimal 0 4 4 2 0 0 0 4 Slight 0 0 2 4 0 0 2 1 Moderate 0 0 0 1 0 0 0 1 Total 0 4 6 7 0 0 2 6 Adhesions/Inflammation/Fib ro sis Minimal 0 3 2 2 0 0 2 1 Slight 0 2 1 3 0 0 0 3 Moderate 0 0 1 1 0 0 0 0 Total 0 5 4 6 0 0 2 4 Number of tissues examined 0 8 7 9 0 0 2 6 

### The incidence and distribution o f all other findings were consistent with the common background seen at these laboratories. The histopathological examinat io n of testes and epididymides fro m the F0 generation animals performed after scheduled treatment period revealed no findings attributable to treatment wi th the test article 2 - bromo- 3,3,3 - trifluoropropene.Huntingdon Life Sciences WAG0015

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### The seminiferous tubules were evaluated with respect to their stage in the spermatogenic cy cle and the integri

ty of the various cell

ty pes present wi

thin the different stages. No significant cell

or stage - specific abnormalit ies were seen in the sect io ns of testes either stained with haematoxylin & eosin (H&E) or periodic acid Schiff (PAS). Evaluat io n of the epididymides did not reveal any changes in leukocy te infil tration, cell populat io ns, or sperm numbers. There were no findings to correlate with the organ weight variat io n reported at necropsy . 3.2.15 Primordial ovarian follicle counts Table 31 and Appendix 28 There were no treatment related effects on the primordial ovarian fo llicle counts. 

## 3.3 F1 litter responses 3.3.1 Signs of offspring App endix 20 At505 ppm

the maj ori

ty of offspring born were noted as being cold on Day 1, wi th those fro m dams where the li

tter was sacrificed early

or died f requent ly being noted as dark in colour or inactive. In surviving offspring, no later signs were generally noted. 3.3.2 Litter size, survival indices and sex ratio Table 20-22 and Appendi ces 16-18 At2 90 0 ppm

the mean number of implantations in the pregnant females was reduced (9.5 com pared wi th 15.9 in the control

group). Only

one female produced a litter, in this female there was low post implantation survival (25%) and a lo w live birth index (33%) such that only one pup was alive on Day 1 post partum . This pup was sacrificed due to poor condit io n on Day 1 post partum. At505 ppm

the mean number of implant ations was slight ly lo wer than the controls. Post implantation survival, the live birth index and the viabilit y index were all lower than the controls, resul

ting in a lower total and live litter size on Day 1 post partum, and lower li tter size by Day 10 p ost partum (mean of 8.3 compared to 14.7 in the control group). In addi tion, a number of offspring were sacrificed due to poor condit io n including reduced activit y and reduced body tem perature. At19 8 ppm there was a slight ly lo wer mean number of implan tations. Post implantation survival was slight ly reduced which resulted in a lower total and live litter size on Day 1 post partum. Group mean survival fro m birth to Day 10 post partum

was lower than the controls; primarily due to lower survival in 2/9 litters. There were considered to be no effects of treatment on the mean sex ratio . Although mean values showed an apparent reduction in the percentage of males per litter between birth and Day 10 post partum, review of the individual data and offspring dying did not show any select ive demise of the males.Huntingdon Life Sciences WAG0015

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### 3.3.3 Bodyweight Figure 5 , Table s 23-24 and Appendix 19 There were no effects on mean male or female pup weight on Day 1 at 198 or 505 ppm . Wi th the longer duration of

gestation at 19 8 and 505 ppm, it might be expected that pup weight would be higher, and this was the case at 19 8 ppm. Bo dyweight gain fro m Day 1 to Day 10 was slight ly lo wer than the controls at 19 8 and 505 ppm. There was only a single female pup weighed at 2 90 0 ppm, the weight was slight ly lo wer than the control (5.5g compared to 6.4g). 3.3.4 Offspring macropathology Appendices 21-22 In offspring that died or were sacrificed prior to Day 10, no milk in the stomach was frequently recorded, especially in those dying in the very early days of lactation. This was mo st apparent at 505 ppm where there were a higher number of deaths. There were no treatment related effects on the offspring sacrificed on Day 10 of lactation.Huntingdon Life Sciences WAG0015

45 

# 4. Discussion Exposure of 2 - bro mo -3,3,3- trifluoropropene to rats was undertaken as an ini ti al screening assessment of the influence on reproductive performance and development fo llo wing administration by inhalat io n fo r a minimum of 15 days before pairing unt il Day 9 af ter the birth of the F1 generat io n, and were sacrificed on Day 10 after the birth of the F1 generat io n . Females were not exposed between Day 20 after mating and Day 4 of l actation inclusive. All females exposed to 2900 ppm and half of those exposed to 505 ppm were killed before Day 10 of lactation as they

f ailed to mate (sacrifi ced on Day 25 af ter the last paring), failed to litter (sacrificed on Day

25 af ter mat ing) , lo st the litter post - partum (sacrificed on the Day of litter loss) or were sacrificed due to poor health . As such, these females did not continue on to Day 10 of lactation. There was an effect on oestrous cy cles of females at 505 and 2900 ppm, as indicated by an increased incidence of “abnormal”. Only 1/10 females at 2900 ppm and 5/10 females at 505 ppm had “normal” cycles. Mating performance was affected at 505 and 2900 ppm with a reduction in the number of pai rings wi th evidence of mat ing, and on the day of mat ing, a reduced presence of copulat io n pl ugs or sperm

wi thin the vaginal smear. The fertilit y index was reduced at 2900 ppm, probably linked wi

th the marked effect on oestrous cycles. There was also a reduced number of implantations evident at sacrifice. Of f emales that showed evidence of mat ing, the delayed pre - coi tal int erval i s considered in part to reflect the reduction in regular oestrous cycles in these groups , as successful mat ings tended to occur at the first oestrous opportunit y af ter pai ring, regardless of whether this was in the “expected” 4 days post pairing (the time of a “normal” oestrous cycle) . The duration of

gestation was extended at 505 and 2900 ppm . Routinely pregnant females com plete parturi

tion on Day

22 af ter mat ing. At 2900 ppm only one o f the six pregnant females littered, on Day

25 af ter mat ing. Two other females were sacrificed on Day 24 after mat ing, the signs indicated this wa s possibly associated with stress invo lved in commencement of parturit io n. The remaining 3 females were sacrificed on Day 25 af ter mat ing as parturit io n had not commenced and there was no indication of this occurring

– these females were found to have been pregnant but had no viable conceptuses at necropsy, it is unclear if these females had shown total litter loss in utero or had commenced parturit io n and the pups born were eaten prior to being observed at the animal room checks. At 505 ppm 9 of the 10 p regnant females produced a litter, although in one case all pups were fo und to be dead at the Day 1 check. The duration of gestation, in f emales at 505 ppm, was extended to between 23 and 25 days. At2900 and 505 ppm, a lo wer number of implantations, coupled wi th reduced in utero survival and reduced post partum viabilit y resul ted in reduced number of offspring surviving to Day 10 post partum; this being a combination o f all o ffspring in the litter dying, or offspring and the dam being sacrificed due to the clinical signs seen. There was a higher incidence of offspring in treated groups being noted as cold, inact ive or unfed. A reduction in the number of sperm in the cauda epidi dymis at 2900 ppm correlated with the lo wer caudal epididymal weight, although the number of sperm/g epididymis was unaffectedHuntingdon Life Sciences WAG0 015

46 

### and there were no changes noted histologically. This also correlated with the lower estimates of sperm present in the vaginal smear of females on the day of copulat io n. At 2900 ppm and generally at 505 ppm, a reduced percentage progressively moti le sperm and sperm

veloci ty and an increased B eat Cross Frequency suggested that whilst there was no reduction in vigour of the sperm, thei

r f orward movement was affected, and an increase in abnormal sperm (mainly breakages) suggested that sperm fragilit y was increased. The reduced body

weight gain and food intake of males at 505 and 2900 ppm and of females at 2900 ppm only was considered an adverse effect of treatment but was not considered sufficient to have been linked to the effect on oestrous cycles or mating performance or the lo wer numbers o f implantations . At 505 ppm 4 females and their o ffspring did not survive to Day 10 post partum . The clinical signs noted during exposure, notably reduced activit y and unresponsiveness, were not unexpected fo llo wing findings on the preliminary range finding study . Clinically the signs had resolved immediately after exposure or before the end of the working day . However, in a concurrent 13 week study in these laboratories [ WAG0014 ] these clinical signs have also been seen and motor activit y mo nit oring in Week 13 showed reduced low and high level act ivit y at similar exposure levels on the morning after exposure. It is uncertain what effect, if any, the reduced activit y had on the mat ing performance. Water int ake was generally higher in females at 505 or 2900 ppm prior to pairing and th roughout gestation. During lactation however, water consumpt io n was lower. The significance of this is unclear but i t may reflect a lower demand on the lactating dams in treated groups due to the lower number of offspring present. No histopathological findings were seen to account for the irregular oestrus cy cles or the prolonged gestation lengths. The examinat io n of the ovaries fro m the F0 generation fe males revealed the presence o f histopathological changes. A reduction in size of corpora lutea was observed in the ovaries of f our f emales exposed to 2900 ppm and one female exposed to 198 ppm. The corpora lutea were qui te small and appeared to be not newly formed. As this change was not co nsistent wi th the increasing exposure level across the treated groups, the presence in a single female exposed to 198 ppm was considered to be incidental. In all cases in the 2900 ppm group, the females wit h s mall corpora lutea were found to be non pregnant; since this finding was not apparent amongst the pregnant females in this group it is mo re likely to relate to their pregnant status and not be a specific effect of treatment. An increased incidence of small prostates was observed at the macroscopic examinat io n. This was consistent with a reduction in prostate weight seen with increasing exposure level. No microscopic correlate for the small prostates could be ident ified. A low incidence of inflammatory

changes was observed in all groups including controls and was considered to be incidental as these t ypes of changes are occasionally seen in rats of this age. Capsular thickening and adhesio ns were observed in the spleen of the majorit y of treated males and occasional

treated females at the macroscopic examinat io n. Most of these macroscopic observations correlated with findings of capsular/subcapsular inflammat io n and/or capsular thickening and/or adhesio ns/infla mmat io n/ fibrosis seen at the microscopic examinat io n and are likely to be related to treatment. However, as no controls and onlyHuntingdon Life Sciences WAG0015

47 

### tissues wi th macroscopic changes were sampled and microscopically examined, concl usions as to the significance of these changes could not be made. The significance of the increased incidence of pale incisors and hair loss of the skin observed clinically and at the macroscopic examinat io n was not further clarified as these tissues are not routi nely examined microscopically. Full evaluation of the spleen capsular thickening and adhesio ns, pale incisors and hairloss are considered to be within the remit of the concurrent 13 week toxicit y study in the rat at the same target exposure concentrations [ WAG0014 ] . It was considered that the effects seen at 505 and 2900 ppm were cons idered adverse. At198 ppm

some similar effects were apparent but the magnitude of the responses tended to be much lower and mo st likely not to be adverse. These not adverse effects included the effects on bodyweight , f

ood intake, water intake, and prob ably the shift to a 5 day oestrous cy cle length (which is st ill considered normal). There was a slightly lower sperm count noted in the vaginal smear on the day of copulation, al

though at terminat io n the epididymal sperm count was not statist ically signif icantly reduced, there was no effect on the epididymal weight and as such any adversit y of this sperm

count is unclear. The significance of the slight shift to a longer duration of

gestation (23 days instead of 22 days) is unclear. At198 ppm, the mean number of implantations was slight ly lo wer than in Controls. Three out of 9 litters showed lower than expected post implantation survival o f the conceptuses (63 - 74%) and two litters showed lower than expected pup viabilit y indices (54 and 82%), and these d ifferences contributed to lower total and live litter sizes than in Controls. Although only a f ew litters were invo lved, the reduction in total and live litter size on Day 1 of lactation fo rmed part of a dose related trend and thus a relationship to treatment was considered likely . In general, treatment affected both the male reproductive system (effects on sperm) and the female reproductive system (oestrous cy cles). However, which sex(es) contributed to the reductions in implantation counts and the pre and post natal survival o f the offspring cannot be determined within the context of this screening study.Huntingdon Life Sciences WAG0015

48 

# 5. Conclusion Exposure of 2 - bro mo -3,3,3- trifluoropropene to rats in this screening study to assess the influence on reproductive performance and develo p me ntf ollo wing administration by inhalat io n at concentrations of 198, 505 or 2900 ppm was associated with adverse effects on male and female toxicit y, and reproductive performance and development at 505 and 2900 ppm. Exposure at 505 and 2900 ppm produced e ffects on oestrous cycles, fert ilit y (2900 ppm only), extended duration of gestation, sperm counts and motilit y, implantation counts, reduced in utero and post partum survival, in the presence of signs of adult toxicit y. In addi tion, at 2900 ppm, reduced size of corpora l utea in the ovaries was noted. Exposure at 198 ppm was associated with slight effects on adult toxicit y and did have slight effects on reproductive performance (a possible shift to longer duration of gestation) and development (slightly lower implantation rate, higher post implantation survival and viabilit y indices leading to lower litter size) as seen at 505 ppm and above. Wit hin this screening study, the no adverse effect level (NOAEL) for reproductive performance and development was co nsidered to lie below 198 ppm .Huntingdon Life Sciences WAG0015

49 

# 6. References 6.1 Necropsy and histology Salewski E (1964). Farbemethode zum makrosc opischen nachweis von implantationsstellen am uterus der Ratte. Naunyn - Schmiedebergs Arch. exp. Pathol. Pharmacol. , 247, 367. 

## 6.2 Statistics Bartlett MS (1937) .Properties of su fficiency and statistical tests. Proceedings of the Royal Society. Series A 160 , 268-282. Cytel (1995). StatXact 3 for Windows: Statistical Software for Exact Nonparametric Inference. Cytel Software Corporation, NC, USA . Dunnett CW (1955). A multiple comparison proce dure for comparing several treatments with a control. Journal of the American Statistical Association 50 , 1096-1121. Dunnett CW (1964). New tables for multip le comparisons with a control. Biometrics 20 , 482-491. Fisher RA (1973). Statistical Methods fo r Research Workers, 14th edn., p.96. Hafner Publishing Company, New York, USA. Huntingdon Life Sciences Study number: WAG0013 2-bromo-3,3,3-trifluoropropene: Dose Range Fi nding Study by Inhalati on Administration to Rats for 2 Weeks. Huntingdon Life Sciences Study number: WAG0014 2-bromo-3,3,3-trifluoropropene: To xicity Study by Inhalation Administration to Rats for 13 Weeks. Huntingdon Life Sciences Study number: WAG0024 2-bromo-3,3,3-trifluoropropene: Dose Range Fi nding Study by Inhalati on Administration to Rats for 2 Weeks. Lipsitz SH, Laird NM, and Harrington DP (1991) . Generalized Estimating Equations for Correlated Binary Data: Using the Odds Ratio as a Measure of Association, Biometrika , 78 , 153-160. Shirley EAC (1977). A non-parametric equivalent of Williams' test for contrasting increasing dose levels of a treatment. Biometrics 33 , 386-389. Steel RGD (1959). A multiple comparison rank sum test: treatments versus control. Biometrics 15 , 560-572. Williams DA (1971). A test for differences between treatment means when several dose levels are compared with a zero dose control. Biometrics 27 , 103-117. Williams DA (1972). The comparison of several dose levels with a zero dose control. Biometrics , 28 , 519-531.50Huntingdon Life SciencesWAG0015

FIGURE 1 Bodyweight - gr oup mean values for males - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900

Pairing commenced Day 1551Huntingdon Life SciencesWAG0015

FIGURE 2 Bodyweight - gr oup mean values for females before pairing - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 290052Huntingdon Life SciencesWAG0015

FIGURE 3 Bodyweight - gr oup mean values for females during gestation - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900

a Mean of females wh ich littered b Mean of all pregnant females53Huntingdon Life SciencesWAG0015

FIGURE 4 Bodyweight - gr oup mean values for females during lactation - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900

Treatment reintroduced Day 5 of lactation54Huntingdon Life SciencesWAG0015

FIGURE 5 Bodyweight - gr oup mean values for offspring - male animals - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 1 98 505 290055Huntingdon Life SciencesWAG0015

FIGURE 5 - continued Bodyweight - gr oup mean values for offspring - female animals - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 290056Huntingdon Life SciencesWAG0015

TABLE 1 Summary of adult performance Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group: 1 2 3 4 Achieved concentration (ppm): 0 198 505 2 900 Males Animals paired: 10 10 10 10 Animals which failed to mate 0 0 0 3 (232, 235, 237) Females Animals paired: 10 10 10 10 Animals which failed to mate 0 0 0 3 (272, 275, 277) Failed to produce a litter (no n - pr egnant) 0 1 (257) 0 1 (280) Number of animals pregnant 10 9 10 6 Failed to produce a litter (total resorption) 0 0 1 (263) 3 (271, 274, 278) Animals sacrificed Day 24 gestation (pregnant) 0 0 0 2 (276, 279) Total litter loss pr e Day 1 of lactation 0 0 1 (26 6 ) 0 Live litter born 10 9 8 1 Total litter loss post - partum 0 0 4 (26 2 a, 26 5 , 267, 270) 1 (273) Litter surviving to Day 10 post partum 10 9 4 0 ( ) Animal number a Includes dam/pups killed for welfare reasons57Huntingdon Life SciencesWAG0015

TABLE 2 Request ID: 284 416 Clinical signs - gr oup distribution of observations for males - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 290 0 Weeks - 1 - 8 Number of animals affected Group/sex: 1M 2M 3M 4M Category Observation Number in group: 10 10 10 10 Coat Hair loss, Cervical 0 0 0 2 Hair loss, Cervical (Left) 0 0 0 1 Hair loss, Head 0 0 1 5 Teeth Abnormal Colour, Pale 0 0 3 1058Huntingdon Life SciencesWAG0015

TABLE 2 - continued Request ID: 284417 Clinical signs - gr oup distribution of observations for females before pairing - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks - 1 - 8 Number of animals affected Group/sex: 1F 2F 3F 4F Category Observation Number in group: 10 10 10 10 Behaviour Aggressive 0 0 1 1 Irritable 1 0 0 0 Vocalisation 1 0 1 2 Build Co nformation Thin 0 0 0 1 Coat Hair loss, Head 1 0 0 0 Piloerection 0 0 0 3 Muscle Reaction Reduced Body Tone 0 0 0 2 Posture Hunched 0 0 0 4 Staining Abnormal Colour, Brown, Head 0 0 0 5 Abnormal Colour, Brown, Muzzle 0 0 0 1 Abnormal Colour, Eye (Left) 0 0 0 1 Abnormal Colour, Eye (Right) 0 0 0 1 Abnormal Colour, Red, Muzzle 0 0 0 1 Teeth Abnormal Colour, Pale 0 0 0 359Huntingdon Life SciencesWAG0015

TABLE 2 - continued Request ID: 284418 Clinical signs - group distribution of observations for females after mating - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Days 0 - 25 Number of animals affected Group/sex: 1F 2F 3F 4F Category Observation Number in group: 10 10 10 7 Behaviour Irritable 0 0 1 0 Un deractive 0 0 0 2 Body temperature Reduced 0 0 0 1 Build Co nformation Thin 0 0 0 1 Coat Hair loss, Forelimb (Right) 0 1 0 0 Hair loss, Forelimbs 1 0 1 0 Hair loss, Head 1 0 1 0 Hair loss, Ventral Body Surface 0 0 1 0 Piloerection 0 0 0 3 Eyelids Partially Closed 0 0 0 1 Muscle Reaction Reduced Body Tone 0 0 0 1 Reduced Body Tone, Whole Body 0 0 0 1 Posture Hunched 0 0 0 1 Staining Abn ormal Colour, Brown, Head 0 1 2 5 Abnormal Colour, Brown, Muzzle 0 0 0 260Huntingdon Life SciencesWAG0015

TABLE 2 - continued Request ID: 284418 Clinical signs - gr oup distribution of observations for females after mating - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Days 0 - 25 Number of animals affected Group/sex: 1F 2F 3F 4F Category Observati on Number in group: 10 10 10 7 Staining Abnormal Colour, Red, Perigenital 0 0 0 1 Teeth Abnormal Colour, Pale 0 0 3 661Huntingdon Life SciencesWAG0015

TABLE 2 - continued Request ID: 284419 Clinical signs - gr oup distribution of observations for females during lactation - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Days 1 - 10 Number of animals affected Group/sex: 1F 2F 3F 4F Category Observation Number in group: 10 9 8 1 Coat Hair loss, Forelimb (Left) 0 1 0 0 Hair loss, Forelimb (Right) 0 1 0 0 Hair loss, Forelimbs 4 3 0 0 Hair loss, Head 1 0 0 0 Hair loss, Hindlimb (Right) 1 1 0 0 Hair loss, Ventral Body Surface 0 3 0 0 Staining Abnormal Colour, Brown, Head 1 2 2 0 Abnormal Colour, Head 0 0 1 0 Teeth Abnormal Colour, Pale 0 0 2 162Huntingdon Life SciencesWAG0015

TABLE 3 Request ID: 284420 Signs associated with dosing - gr oup distribution of observations for males - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Day Group/sex and number of animals showing sign Sign Number 1M 2M 3M 4M Behaviour Un deractive 1 0 10 10 10 2 0 10 10 10 3 0 1 0 10 10 4 0 10 10 10 5 0 10 10 10 6 0 10 10 10 7 0 10 10 10 11 0 0 10 10 14 0 0 10 10 18 0 0 10 10 21 0 0 10 10 25 0 10 10 10 28 0 10 10 10 32 0 0 10 10 35 0 0 10 10 40 0 0 10 10 47 0 0 10 10 Signs recorded during exposure on a group basis, all animals in the group assumed to have the same sign63Huntingdon Life SciencesWAG0015

TABLE 3 - continued Request ID: 284420 Signs associated with dosing - gr oup distribution of observations for males - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Day Group/sex and number of animals showing sign Sign Number 1M 2M 3M 4M Behaviour (continued...) Unresponsive 1 0 10 10 10 2 0 0 10 10 3 0 10 10 0 4 0 0 0 10 5 0 0 0 10 6 0 0 10 10 7 0 0 10 10 11 0 0 0 10 14 0 0 0 10 18 0 0 10 10 21 0 0 10 10 25 0 0 10 10 32 0 0 0 10 40 0 0 0 10 47 0 0 0 10 Signs recorded during exposure on a group basis, all animals in the group assumed to have the same sign64Huntingdon Life SciencesWAG0015

TABLE 3 - continued Request ID: 284420 Signs associated with dosing - gr oup distribution of observations for males - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Day Group/sex and number of animals showing sign Sign Number 1M 2M 3M 4M Br eathing Shallow 1 0 0 0 10 2 0 0 0 10 3 0 0 0 10 4 0 0 0 10 5 0 0 0 10 6 0 0 0 10 7 0 0 0 10 14 0 0 10 10 Slow 25 0 0 10 10 28 0 0 0 10 35 0 0 10 10 40 0 0 0 10 Signs recorded during exposure on a group basis, all animals in the group assumed to have the same sign65Huntingdon Life SciencesWAG0015

TABLE 3 - continued Request ID: 284420 Signs associated with dosing - gr oup distribution of observations for males - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - t rifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Day Group/sex and number of animals showing sign Sign Number 1M 2M 3M 4M Coat Piloerection 1 0 0 10 10 2 0 10 10 1 0 3 0 10 10 10 4 0 10 10 10 5 0 10 10 10 6 0 10 10 10 7 0 10 10 10 14 0 10 10 10 18 0 0 10 10 21 0 0 10 10 25 0 10 0 0 28 0 10 10 10 32 0 0 10 10 35 0 1 10 0 40 0 0 10 10 47 0 0 10 10 Eye (Left) Lachry mation 1 0 0 10 0 Eye (Right) Lachry mation 1 0 0 10 0 Signs recorded during exposure on a group basis, all animals in the group assumed to have the same sign66Huntingdon Life SciencesWAG0015

TABLE 3 - continued Request ID: 284420 Signs associated with dosing - gr oup distribution of observations for males - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Day Grou p/sex and number of animals showing sign Sign Number 1M 2M 3M 4M Eyelids (Left) Partially Closed 1 0 0 10 10 3 0 0 10 10 4 0 0 10 10 5 0 0 10 10 6 0 0 0 10 7 0 0 0 10 11 0 0 10 10 14 0 10 10 10 18 0 0 0 10 21 0 0 0 10 25 0 0 0 10 28 0 0 0 10 35 0 0 10 10 40 0 0 0 10 47 0 0 0 10 Signs recorded during exposure on a group basis, all animals in the group assumed to have the same sign67Huntingdon Life SciencesWAG0015

TABLE 3 - continued Request ID: 284420 Signs associated with dosing - gr oup distribution of observations for males - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Day Group/sex and number of animals showing sign Sign Number 1M 2M 3M 4M Eyelids (Right) Partially Closed 1 0 0 10 10 3 0 0 10 10 4 0 0 10 10 5 0 0 10 10 6 0 0 0 10 7 0 0 0 10 11 0 0 10 10 14 0 10 10 10 18 0 0 0 10 21 0 0 0 10 25 0 0 0 10 28 0 0 0 10 35 0 0 10 10 40 0 0 0 10 47 0 0 0 10 Signs recorded during exposure on a group basis, all animals in the group assumed to have the same sign68Huntingdon Life SciencesWAG0015

TABLE 3 - continued Request ID: 284421 Sign s associated with dosing - gr oup distribution of observations for females before pairing - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Day Group/sex and number of animals showing sign Sign Number 1F 2F 3F 4F Behaviour Un deractive 1 0 10 10 10 2 0 10 10 10 3 0 10 10 10 4 0 10 10 10 5 0 10 10 10 6 0 10 10 10 7 0 10 10 10 8 0 0 0 3 11 0 0 10 10 14 0 0 10 10 18 0 0 9 8 21 0 0 1 6 25 0 1 0 4 28 0 0 0 3 32 0 0 0 3 35 0 0 0 3 40 0 0 0 3 47 0 0 0 3 Signs recorded during exposure on a group basis, all animals in the group assumed to have th e same sign69Huntingdon Life SciencesWAG0015

TABLE 3 - continued Request ID: 284421 Signs associated with dosing - gr oup distribution of observations for females before pairing - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Day Group/sex and number of animals showing sign Sign Number 1F 2F 3F 4F Behaviour (continued...) Unresponsive 1 0 10 10 10 2 0 0 10 10 3 0 10 10 0 4 0 0 0 10 5 0 0 0 10 6 0 0 10 10 7 0 0 10 10 8 0 0 0 3 11 0 0 0 10 14 0 0 0 10 18 0 0 9 8 21 0 0 1 6 25 0 0 0 4 32 0 0 0 3 40 0 0 0 3 47 0 0 0 3 Signs recorded during exposure on a group basis, all an imals in the group assumed to have the same sign70Huntingdon Life SciencesWAG0015

TABLE 3 - continued Request ID: 284421 Signs associated with dosing - gr oup distribution of observations for females before pairing - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Day Group/sex and number of animals showing sign Sign Number 1F 2F 3F 4F Br eathing Shallow 1 0 0 0 10 2 0 0 0 10 3 0 0 0 10 4 0 0 0 10 5 0 0 0 10 6 0 0 0 10 7 0 0 0 10 14 0 0 10 10 Slow 25 0 0 0 4 28 0 0 0 3 35 0 0 0 3 40 0 0 0 3 Signs recorded during exposure on a group basis, all animals in the group assumed to have the same sign71Huntingdon Life SciencesWAG0015

TABLE 3 - continued Request ID: 284421 Signs associated with dosing - gr oup distribution of observations for females before pairing - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

t rifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Day Group/sex and number of animals showing sign Sign Number 1F 2F 3F 4F Coat Piloerection 1 0 0 10 10 2 0 10 10 10 3 0 10 10 10 4 0 10 1 0 10 5 0 10 10 10 6 0 10 10 10 7 0 10 10 10 8 0 0 0 3 11 0 10 0 0 14 0 10 10 10 18 0 0 9 8 21 0 0 1 6 25 0 1 0 0 28 0 0 0 3 32 0 0 0 3 40 0 0 0 3 47 0 0 0 3 Eye (Left) Lachry mation 1 0 0 10 0 Signs recorded during exposure on a group basis, all animals in the group assumed to have the same sign72Huntingdon Life SciencesWAG0015

TABLE 3 - continued Request ID: 284421 Signs associated with dosing - gr oup distribution of observations for females before pairing - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Day Group/sex and number of animals showing sign Sign Number 1F 2F 3F 4F Eye (Right) Lachry mation 1 0 0 10 0 Eyelids (Left) Partially Closed 1 0 0 10 10 3 0 0 10 10 4 0 0 10 10 5 0 0 10 10 6 0 0 0 10 7 0 0 0 10 8 0 0 0 3 11 0 0 10 10 14 0 10 10 10 18 0 0 0 8 2 1 0 0 0 6 25 0 0 0 4 28 0 0 0 3 35 0 0 0 3 40 0 0 0 3 47 0 0 0 3 Signs recorded during exposure on a group basis, all animals in the group assumed to have the same sign73Huntingdon Life SciencesWAG0015

TABLE 3 - continued Request ID: 284421 Signs associated with dosing - gr oup distribution of observations for females before pairing - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Day Group/sex and number of animals showing sign Sign Number 1F 2F 3F 4F Eyelids (Right) Partially Closed 1 0 0 10 10 3 0 0 10 10 4 0 0 10 10 5 0 0 10 10 6 0 0 0 10 7 0 0 0 10 8 0 0 0 3 11 0 0 10 10 14 0 10 10 10 18 0 0 0 8 21 0 0 0 6 25 0 0 0 4 28 0 0 0 3 35 0 0 0 3 40 0 0 0 3 47 0 0 0 3 Posture Hunched 8 0 0 0 3 Staining Other Colour (see comment) 8 0 0 0 3 Signs recorded during exposure on a group basis, all animals in the group assumed to have the same sign74Huntingdon Life SciencesWAG0015

TABLE 3 - continued Request ID: 284422 Signs associated with dosing - gr oup distribution of observations for females after mating - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Day Group/sex and number of animals showing sign Sign Number 1F 2F 3F 4F Behaviour Un deractive 0 0 3 6 5 7 0 5 10 7 14 0 3 10 7 19 0 0 10 7 Unresponsive 0 0 0 2 5 7 0 0 6 2 14 0 0 0 5 19 0 0 1 2 Br eathing Shallow 0 0 0 1 0 Slow 0 0 0 5 2 7 0 0 7 6 14 0 0 5 2 19 0 0 10 6 S igns recorded during exposure on a group basis, all animals in the group assumed to have the same sign75Huntingdon Life SciencesWAG0015

TABLE 3 - continued Request ID: 284422 Signs associated with dosing - gr oup distribution of observations for females after mating - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Day Group/sex and number of animals showing sign Sign Number 1F 2F 3F 4F Coat Piloerection 0 0 3 1 4 7 0 9 3 3 14 0 1 7 6 19 0 5 5 3 Eyelids (Left) Partially Closed 0 0 0 5 5 7 0 0 5 7 14 0 0 5 3 19 0 0 8 6 Eyelids (Right) Partially Closed 0 0 0 5 5 7 0 0 5 7 14 0 0 5 3 19 0 0 8 6 Muscle Reaction Reduced Body Tone 14 0 0 0 1 Posture Hunched 14 0 0 0 1 Signs recorded during exposure on a group basis, all animals in the group assumed to have the same sign76Huntingdon Life SciencesWAG0015

TABLE 3 - continued Request ID: 284423 Signs associated with dosing - gr oup distribution of observations for females during lactation - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3, 3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Day Group/sex and number of animals showing sign Sign Number 1F 2F 3F 4F Behaviour Un deractive 5 0 0 2 - 9 0 0 4 - Coat Piloerection 5 0 0 2 - Signs recorded during exposure on a group basis, all animals in the group assumed to have the same sign77Huntingdon Life SciencesWAG0015

TABLE 4 Request ID: 284760 Bodyweight and bodyweight change - gr oup mean values for males - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Day Day Day Day Day Day Day Day Day Day Day Day /Sex - 7 1 8 11 15 22 29 33 36 39 43 46 Statistical test: Wi Wi Wi Wi Wi Wi Wi Wi Wi Wi Wi Wi 1 M Mean 259 329 380 395 420 444 471 487 498 503 517 528

SD 7.7 9.4 13.4 17.8 21.0 26.8 26.4 29.5 30.6 30.3 29.9 29.5 N 10 10 10 10 10 10 10 10 10 1 0 10 10 2 M Mean 254 319 361* 375* 393* 420 443* 452* 460* 465* 469** 476**

SD 9.4 12.8 27.2 33.6 39.5 44.1 47.6 48.8 52.1 51.7 49.9 52.1 N 10 10 10 10 10 10 10 10 10 10 10 10 3 M Mean 256 322 349** 357** 376** 398** 415** 426** 434** 437** 441** 449**

SD 6.6 8.0 12.3 14.7 16.8 19.5 20.3 22.2 22.3 22.8 26.9 27.3 N 10 10 10 10 10 10 10 10 10 10 10 10 4 M Mean 253 325 342** 355** 368** 386** 406** 409** 414** 415** 419** 421**

SD 9.4 12.1 16.0 17.2 20.4 19.0 21.7 24.0 22.5 26.1 26.5 30.9 N 10 10 10 10 10 10 10 10 10 10 10 10 Pairing commenced Day 1578Huntingdon Life SciencesWAG0015

TABLE 4 - continued Request ID: 284760 Bodyweight and bodyweight change - gr oup mean values for males - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Day Change As % of Change As % of Change As % of /Sex 50 1 - 15 Control 15 - 50 Control 1 - 50 Control Statistical test: Wi Wi Wi Wi 1 M Mean 542 91 - 121 - 212 -

SD 33.0 15.4 19.9 27.9 N 10 10 10 10 2 M Mean 483** 74 81 90** 74 164** 77

SD 52.0 30.6 17.4 44.0 N 10 10 10 10 3 M Mean 455** 54** 60 79** 65 133** 63

SD 27.4 13.7 15.4 22.1 N 10 10 10 10 4 M Mean 424** 43** 47 56** 46 99** 47

SD 31.9 14.3 19.1 29.3 N 10 10 10 10 Pairing commenced Day 1579Huntingdon Life SciencesWAG0015

TABLE 5 Request ID: 284761 Bodyweight and bodyweight change - gr oup mean values for females before pairing - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Day Day Day Day Day Day Day Day Day Day Day Day /Sex - 7 1 8 11 15 22 29 33 36 39 43 46 Statistical test: Wi Wi Wi Wi Wi 1 F Mean 202 225 237 239 245

SD 9.7 11.5 12.1 11.2 10.8 N 10 10 10 10 10 2 F Mean 205 227 239 244 252

SD 6.2 5.7 9.0 9. 3 10.9 N 10 10 10 10 10 3 F Mean 202 225 232 236 244

SD 6.2 9.5 7.8 9.9 10.4 N 10 10 10 10 10 4 F Mean 207 228 227* 231 232** 234 240 245 254 245 249 251

SD 8.2 10.8 10.8 6.6 6.6 10.5 5.2 2.9 2.9 2.8 5.2 3.4 N 10 10 10 10 10 5 3 3 3 3 3 3 Pairing commenced Day 1580Huntingdon Life SciencesWAG0015

TABLE 5 - continued Request ID: 284761 Bodyweight and bodyweight change - gr oup mean values for females before pairing - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropr opene Exposure level (ppm) : 0 198 505 2900 Group Day Day Change As % of /Sex 50 53 1 - 15 Control Statistical test: Wi 1 F Mean 20 -

SD 7.1 N 10 2 F Mean 24 121

SD 10.3 N 10 3 F Mean 19 95

SD 6.5 N 10 4 F Mean 250 253 4** 19

SD 7.9 9.8 14.7 N 3 3 10 Pairing commenced Day 1581Huntingdon Life SciencesWAG0015

TABLE 6 Request ID : 284780 Bodyweight and bodyweight change - gr oup mean values for females during gestation - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposur e level (ppm) : 0 198 505 2900 Group Day Day Day Day Day Day Day Change As % of Change As % of /Sex 0 3 7 10 14 17 20 0 - 10 Control 10 - 20 Control Statistical test: Wi Wi Wi lWi Wi Wi Wi Wi Wi 1 F Mean 256 273 294 304 327 360 410 49 - 106 -

SD 12.4 14.2 18.4 14.8 19.8 21.9 26.2 8.3 14.4 N 10 10 10 10 10 10 10 10 10 2 F Mean 255 272 286 296 317 347 397 41 85 100 95

S D 13.0 13.4 17.4 23.0 23.1 28.8 38.5 15.5 26.4 N 9 9 9 9 9 9 9 9 9 3 F Mean 249 265 277* 287* 312 333* 377* 38 78 90 85

SD 9.9 8.7 9.8 6.8 11.9 15.4 22.7 8.6 18.7 N 9 9 9 9 9 9 9 9 9 4 Fa Mean 227 234 239 248 272 281 316 21 43 68 64

N 1 1 1 1 1 1 1 1 1 4 Fb Mean 239 254 260 272 285 293 305 33 67 33 31

SD 15.3 10.1 14.5 12.9 14.3 13.3 17.0 12.4 26.2 N 6 6 6 6 6 6 6 6 6

a Mean of females wh ich littered , not included in statistical evaluation b Mean of all pregnant females , not included in statistical evaluation Treatment withdrawn after Day 19 of gestation82Huntingdon Life SciencesWAG0015

TABLE 6 - continued Request ID: 284780 Bodyweight and bodyweight change - gr oup mean values for females during gestation - F0 generation (g) Group : 1 2 3 4 Compound : Con trol 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Change As % of /Sex 0 - 20 Control Statistical test: Wi 1 F Mean 154 -

SD 18.4 N 10 2 F Mean 142 92

SD 27.7 N 9 3 F Mean 128* 83

SD 16.3 N 9 4 Fa Mean 89 58

N 1 4 Fb Mean 66 43

SD 30.1 N 6

a Mean of females wh ich littered , not included in statistical evaluation b Mean of all pregnant females, not included in statistical evaluation Treatment withdrawn after Day 19 of gestation83Huntingdon Life SciencesWAG0015

TABLE 7 Request ID: 284763 Bodyweight and bodyweight change - gr oup mean values for females during lactation - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Day Day Day Change As % of Change As % of Change As % of /Sex 1 5 10 1 - 5 Control 5 - 10 Control 1 - 10 Control Statistical test: Wi Wi Wi Wi Wi Wi 1 F Mean 304 322 341 18 - 20 - 37 -

SD 19.4 26.2 23.2 12.9 12.5 14.7 N 10 10 10 10 10 10 2 F Mean 300 311 322 11 61 11 57 22* 59

SD 20.0 23.2 27.1 9.2 10.5 9.6 N 9 9 9 9 9 9 3 F Mean 292 299 297** 7 42 - 2** - 5** 13

SD 12.3 6.8 15.2 6.7 13.7 15.8 N 4 4 4 4 4 4 Treatment reintroduced Day 5 of lactation84Huntingdon Life SciencesWAG0015

TABLE 8 Request ID: 284799 Food consumption - gr oup mean values for males and females before pairing - F0 generation (g/animal/week) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Week Week As % of Week As % of Mean As % of /Sex -1 1 Control 2 Control 1-2 Control 1M Mean 207 220 - 212 - 216 -

SD 8.3 10.9 10.3 10.6 N 2 2 2 2 2M Mean 198 191 87 190 90 191 88

SD 3.1 9.5 19.3 14.4 N 2 2 2 2 3M Mean 200 162 74 165 78 164 76

SD 3.8 7.9 1.9 4.9 N 2 2 2 2 4M Mean 204 149 68 179 84 164 76

SD 8.9 5.4 8.6 7.0 N 2 2 2 2 

### Week 2 consumption based on Days 1 - 6 of the week and adjusted accordingly to give a representative weekly value.85Huntingdon Life SciencesWAG0015

TAB LE 8 - continued Request ID: 284799 Food consumption - gr oup mean values for males and females before pairing - F0 generation (g/animal/week) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Week Week As % of Week As % of Mean As % of /Sex -1 1 Control 2 Control 1-2 Control 1F Mean 137 133 - 130 - 132 -

SD 2.6 2.1 1.6 1.9 N 2 2 2 2 2 F Mean 137 117 88 124 95 121 92

SD 0.1 0.4 2.7 1.6 N 2 2 2 2 3F Mean 136 101 76 114 88 108 82

SD 1.1 0.5 1.5 0.5 N 2 2 2 2 4F Mean 138 86 65 98 75 92 70

SD 8.9 5.7 4.1 4.9 N 2 2 2 286Huntingdon Life SciencesWAG0015

TABLE 9 Request ID: 284781 Food consumption - gr oup mean values for females during gestation - F0 generation (g/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Day Day Day Day Day Day Mean As % of /Sex 0-2 3-6 7-9 10-13 14-16 17-19 0-19 Control Statistical test: Wi Wi Wi Wi Wi Wi Wi 1F Mean 24 26 26 27 28 29 27 -

SD 3.5 3.3 2.9 2.9 3.1 3.5 2.6 N 10 10 10 10 10 10 10 2F Mean 22 21** 21** 25* 25 30 24* 90

SD 2.4 2.9 3.3 2.3 3.6 3.2 2.1 N 9 9 9 9 9 9 9 3F Mean 21 22** 22** 24** 24** 29 24** 89

SD 1.7 1.9 2.6 1.9 2.2 1.1 1.5 N 9 9 9 9 9 9 9 4Fa Mean 15 16 17 20 21 29 20 74

N 1 1 1 1 1 1 1 4Fb Mean 19 19 20 21 21 24 21 78

SD 2.0 2.8 1.6 2.2 3.3 3.3 1.4 N 6 6 6 6 6 6 6

a Mean of females wh ich littered , not included in statistical evaluation b Mean of all pregnant females, not included in statistical evaluation87Huntingdon Life SciencesWAG0015

TABLE 10 Request ID: 285999 Food consumption - gr oup mean values for females during lactation - F0 generation (g/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3, 3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Day Day Mean As % of /Sex 1-4 5-9 1-9 Control Statistical test: Wi Wi Wi 1F Mean 40 56 49 -

SD 4.4 5.1 4.6 N 10 10 10 2F Mean 31** 39** 35** 73

SD 5.6 6.2 5.7 N 9 9 9 3F Mean 28** 30** 29** 60

SD 5.0 4.9 4.9 N 4 4 4 Treatment reintroduced Day 5 of lactation88Huntingdon Life SciencesWAG0015

TABLE 11 Water consumption - gr oup mean values for males and before pairing for females - F0 generation (mL/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Anima l Week Week Week Week /Sex Number 2 5 6 $ 7 1M Mean 45 41 42 42 SD 3.5 1.4 4.2 2.1 N 2 2 2 2 2M Mean 43 38 40 37 SD 8.5 6.4 7.8 5.7 N 2 2 2 2 3M Mean 44 40 42 40 SD 2.8 0.0 0.0 0.7 N 2 2 2 2 4M Mean 47 39 39 39 SD 0.0 2.1 4.9 4.2 N 2 2 2 2 Calculated for 3 days/week $ Calculated over a one day period Week 6 in error, Week 7 consumption recorded over a 6 day period to provide additional data89Huntingdon Life SciencesWAG0015

TABLE 11 - continued Water consumption - gr oup mean values for males and before pairing for females - F0 generation (mL/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Week As % of /Sex Number 2 Control 1F Mean 29 - SD 0.0 N 2 2F Mean 34 117 SD 0.0 N 2 3F Mean 46 159 SD 2.1 N 2 4F Mean 50 172 SD 9.9 N 2 Calculated for 3 days/week90Huntingdon Life SciencesWAG0015

TABLE 12 Water consumptio

n - gr oup mean values for females during gestation - F0 generation (mL/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Day Day Day Day Day Day Day Day Day Day Day Day Day /Sex 0 1 2 3 4 5 6 7 8 9 10 11 12 1 F Mean 51 67 62 52 53 58 56 58 58 61 68 63 64 SD 18.3 10.7 10.9 13.2 9.2 11.2 9.0 9.2 8.9 10.3 10.5 9.9 10.5 N 10 10 10 10 10 10 10 10 10 10 10 10 9 2 F Mean 43 56 66 67 61 63 64 69 67 68 74 72 74 SD 13.4 17.0 22.3 20.2 13.9 15.7 14.2 10.6 13.3 12.5 10.3 13.1 14.1 N 9 9 9 9 9 9 9 9 9 9 9 9 9 3 F Mean 54 63 67 62 59 65 62 63 64 69 80 78 73 SD 13 .7 15.5 15.5 17.6 15.7 16.5 13.2 11.2 12.1 13.5 8.4 9.4 16.7 N 9 9 9 9 9 9 9 9 9 9 9 9 9 4 F a Mean 34 42 75 80 68 51 70 73 68 74 57 79 43

N 1 1 1 1 1 1 1 1 1 1 1 1 1 4 Fb Mean 60 63 74 70 62 63 65 67 63 68 73 84 69 SD 14.5 19.9 11.9 13.3 8.9 12.8 10.7 14.5 12.4 16.3 12.8 11.8 16.8 N 6 6 6 6 6 6 6 6 6 6 6 6 5

a Mean of females wh ich littered , not included in statistical evaluation b Mean of all pregnant females, not included in statistical evaluation91Huntingdon Life SciencesWAG0015

TABLE 12 - continued Water consumption - gr oup mean values for females during gestation - F0 generation (mL/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropr opene Exposure level (ppm) : 0 198 505 2900 Group Day Day Day Day Day Day Days Days Days Days Days Days /Sex 13 14 15 16 17 18 0 - 2 3 - 5 6 - 8 9 - 11 12 - 15 16 - 17 1 F Mean 69 65 68 68 72 74 60 54 57 64 67 70 SD 8.6 7.6 10.4 9.8 9.6 9.0 10.5 8.4 7.6 7.6 7.4 7.7 N 10 10 10 10 10 10 10 10 10 10 10 10 2 F Mean 77 78 77 82 75 67 55 64 67 71 76 79 SD 11.0 11.3 13.7 16.2 12.9 10.1 15.7 12.6 10.5 8.2 11.0 12.0 N 9 9 9 9 9 9 9 9 9 9 9 9 3 F Mean 87 82 86 91 91 89 61 62 63 76 82 91 SD 14.2 16.6 20.4 5.7 11.2 11.1 13.4 14.5 11.8 6.9 15.5 8.2 N 9 9 9 9 9 9 9 9 9 9 9 9 4 F a Mean 44 47 74 53 49 47 50 66 70 70 52 51

N 1 1 1 1 1 1 1 1 1 1 1 1 4 Fb Mean 74 72 90 85 70 74 66 65 65 75 77 77 SD 19.9 16.4 14.0 18.2 21.4 22.5 10.1 9.6 11.1 6.3 14.2 18.2 N 6 6 6 6 6 6 6 6 6 6 6 6

a Mean of females wh ich littered , not included in statistical evaluation b Mean of all pregnant females, not included in statistical evaluation92Huntingdon Life SciencesWAG0015

TABLE 13 Water consumption - gr oup mean values for females during lactation - F0 generation (mL/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day Day Day Day Day Day Day Day Days Days /Sex Number 1 2 3 4 5 6 7 8 9 1 - 3 4 - 9 1 F Mean 71 75 63 80 88 89 90 105 100 69 92 SD 29.2 30.8 11.8 21.4 13.8 12.2 13.0 10. 1 17.9 10.6 9.3 N 10 10 10 10 10 10 10 10 10 10 10 2 F Mean 63 76 57 52 67 82 76 84 93 65 76 SD 39.8 37.1 23.1 12.9 11.6 66.8 12.5 20.2 18.4 11.8 16.3 N 9 9 9 9 9 9 9 9 9 9 9 3 F Mean 33 44 54 64 53 70 79 86 74 44 71 SD 6.3 8.5 13.1 12.3 13.6 27.6 12.1 8.4 19.5 8.4 11.3 N 4 4 4 4 4 4 4 4 4 4 493Huntingdon Life SciencesWAG0015

TABLE 14 Oestrous cycles - incidence of females with specific cycle lengths - gr oup values - F0 generation Group : 1 2 3 4 Compound : C ontrol 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900

Number of Cy cle length (days) Group animals 2 3 4 5 6 - 10 11 - 20 Statistical test: Ca 1 10 n 0 0 9 2 0 0 (%) (90) (20) 2** 10 n 0 0 6 9 0 0 (%) (60) (90) 3** 10 n 0 0 2 8 4 1 (%) (20) (80) (40) (10) 4** 10 n 1 0 1 5 7 5 (%) (10) (10) (50) (70) (50)

n Number of females with at least one cycle in category (each f emale may

have cycles in more than one category)94Huntingdon Life SciencesWAG0015

TABLE 14 - continued Oestrous cycles - incidence of specific cycle lengths - gr oup values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

t rifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900

Number Number of of Cy cle length (days) Group animals cycles 2 3 4 5 6 - 10 11 - 20 Statistical test: Lt 1 10 39 n 0 0 32 7 0 0 (%) (82) (18) 2** 10 33 n 0 0 10 23 0 0 (%) (30) (70) 3** 10 33 n 0 0 6 20 6 1 (%) (18) (61) (18) (3) 4** 10 28 n 1 0 1 8 13 5 (%) (4) (4) (29) (46) (18)

n Number of cycles in category95Huntingdon Life SciencesWAG0015

TABLE 15 Pre - coital interval - gr oup values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900

Number of Pre - coital interval (days) Group animals 1 - 4 5 - 8 9 - 12 13 - 14 Statistical test: Lt 1 10 n 9 1 0 0 (%) (90) (10) 2 10 n 9 0 1 0 (%) (90) (10) 3 10 n 7 3 0 0 (%) (70) (30) 4 7 n 4 2 1 0 (%) (57) (29) (14)96Huntingdon Life SciencesWAG0015

TABLE 16 Number of copulation plugs at mating - gr oup values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900

Number of Number of copulation plugs Group animals 1 2 3 4 - 6 Statistical test: Lt 1 10 n 0 0 2 8 (%) (20) (80) 2 10 n 1 1 1 7 (%) (10) (10) (10) (70) 3* 10 n 1 1 6 2 (%) (10) (10) (60) (20) 4* 7 n 0 2 3 2 (%) ( 29 ) (43) (29) Historical control data a

288 % 0 .4 7.6 15.6 72.7 a From 29 studies of similar type and age at pairing97Huntingdon Life SciencesWAG0015

TABLE 17 Sperm count estimates from vaginal smears at mating - gr oup values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900

Number of Sperm count category  Group animals 0 1 2 3 4 Statistical test: Lt 1 10 n 1 0 1 2 6 (%) (10) (10) (20) (60) 2 10 n 0 3 2 4 1 (%) (30) (20) (40) (10) 3 10 n 4 1 1 2 2 (%) (40) (10) (10) (20) (20) 4 7 n 0 0 2 4 1 (%) (29) (57) (14)

 0 No sperm 1 Occasional sperm 2 Continuous few sperm 3 Many scattered sperm 4 Solid masses of sperm98Huntingdon Life SciencesWAG0015

TABLE 18 Mating performance and fertility - group values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Number and Number Number achieving Percentage Conception Fertility sex paired mating pregnancy mating rate (%) index (%) Statistical test: Ca Ca Ca 1M 10 10 10 100 100 100 2M 10 10 9 100 90 90 3M 10 10 10 100 100 100 4M 10 7 6 70* 86 60* St atistical test: Ca Ca Ca 1F 10 10 10 100 100 100 2F 10 10 9 100 90 90 3F 10 10 10 100 100 100 4F 10 7 6 70* 86 60*99Huntingdon Life SciencesWAG0015

TABLE 19 Gestation length and gestation index - gr oup values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900

Number of Number of Gestation pregnant Gestation length (days) live litters index Group animals 22 22.5 23 2 3.5 24 24.5 25 25.5 born (%) Statistical test: Lt Ca 1 10 n 6 3 1 0 0 0 0 0 10 100

(%) (60) (30) (10) 2** 9 n 0 0 4 5 0 0 0 0 9 100

(%) (44) (56) 3** 10 n 0 0 1 3 0 2 1 2 9 90

(%) (11) (33) (22) (11) (22) 4** 6 n 0 0 0 0 0 0 0 1 1 17**

(%) (100) Historical control data a

255 % 32 41 25 1 0 0 0 0 100 a From 26 studies of similar type and age at pairing100Huntingdon Life SciencesWAG0015

TABLE 20 Litter size - gr oup mean values for litters that survived t o Day 10 of lactation - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Implantations Total litter size Live litter size on Day Day 1 1 5 10 Statistical test: Wi Sh Wi Wi Wi 1 Mean 15.9 15.0 14.9 14.8 14.7 SD 1.3 1.2 1.2 1.4 1.3 n 10 10 10 10 10 2 Mean 14.1 12.2 * 11.8 ** 10.6 ** 10.6 ** SD 3.1 3.3 3.1 3.3 3.3 n 9 9 9 9 9 3 Mean a 15.5 11.8 ** 10.8 ** 8.5 ** 8.3 ** SD 1.7 1.0 1.9 2.1 2.1 n 4 4 4 4 4 Meanb 14.1 8.9 ** 11.4 * 8.5 ** 8.3 ** SD 2.6 4.3 2.2 2.1 2.1 n 9 9 5 4 4 4 Mean a 9.5

n 6 Meanb 12.0 3.0 1.0 n 1 # 1 1 # Implantati ons o

nly recorded for one female, not included in statistical evaluation. Group 4 not included in statistical analysis. a Females surviving to Day 10 post partum b All pregnant females/females that littered where applicable101Huntingdon Life SciencesWAG0015

TABLE 21 Offspring surviva l indices - gr oup mean values for litters that survived to Day 10 of lactation - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Post implantation Live birth Viability survival index index index (%) (%) (%) Statistical test: Wi Sh Sh 1 Mean 94.5 99.4 98.6

n 10 10 10 2 Mean 86.0 96.8 90.3

n 9 9 9 3 Mean a 76.8 * 91.3 78.2 *

n 4 4 4 Meanb 56.5** 51.7** 62.6** n 10 9 5 4 Mean a 25.0 33.3

n 1# 1# Meanb 4.2** n 6 # Implantations only recorded for one female , not included in statistical evaluation a Females surviving to Day 10 post partum b All pregnant females/females that littered where applicable102Huntingdon Life SciencesWAG0015

TABLE 22 Sex ratio - gr oup mean values for litters that survived to Day 10 of lactation - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Total on Day Live on Day 1 1 5 10 M F %M M F %M M F %M M F %M Statistical test: Wa Wa Wa Wa 1 Mean 7.3 7.7 48.2 7.2 7.7 48.0 7.2 7.6 48.2 7.2 7.5 48.5

SD 2.5 2.1 15.5 2.3 2.1 15.2 2.3 2.0 14.8 2.3 2.0 15.0 n 10 10 10 10 10 10 10 10 10 10 10 10 2 Mean 6.3 5.8 51.2 6.1 5.7 50.8 5.3 5.2 49.2 5.3 5.2 49.2

SD 2.6 1.9 12.2 2.5 1.9 13.4 2.4 1.9 12.3 2.4 1.9 12.3 n 9 9 9 9 9 9 9 9 9 9 9 9 3 Mean 5.8 5.0 53.4 5.8 5.0 53.4 4.0 4.5 45.1 4.0 4.3 46.1

SD 2.1 2.0 15.2 2.1 2.0 15.2 2.2 1.0 14.7 2.2 0.5 13.7 n 4 4 4 4 4 4 4 4 4 4 4 4103Huntingdon Life SciencesWAG0015

TABLE 23 Bodyweight and bodyweight change - gr oup mean values (g) for male offspring from litters surviving to Day 10 of lactation - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 29 00 Group Day of age Change 1 5 10 1 to 5 1 to 10 Statistical test: Wi Wi Wi Wi Wi 1 Mean 6.8 10.9 17.5 4.1 10.7

SD 0.3 0.6 1.0 0. 4 0.9 n 10 10 10 10 10 2 Mean 7.4 11.1 17.0 3.8 9.6

SD 0.6 1.0 1.2 0. 6 0.8 n 9 9 9 9 9 3 Mean 6.9 10.6 16.0 3.7 9.1 *

SD 0.7 1.8 2.7 1. 3 2.2 n 4 4 4 4 4104Huntingdon Life SciencesWAG0015

TABLE 24 Bodyweight and bodyweight change - gr oup mean values (g) for female offspring from litters surviving to Day 10 of lactation - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Day of age Change 1 5 10 1 to 5 1 to 10 Statistical test: Du Wi Wi Sh Wi 1 M ean 6.4 10.4 16.7 4.0 10.3

SD 0.4 0.6 1.1 0.3 0.9 n 10 10 10 10 10 2 Mean 6.9 ** 10.9 16.2 3.9 9.3

SD 0.4 1.0 1.6 0.7 1. 4 n 9 9 9 9 9 3 Mean 6.2 9.7 15.1 3.4 8.8

SD 0.2 1.5 2.5 1. 4 2.4 n 4 4 4 4 4 4 @ Mean 5 .5

n 1 @ Not mean value, presented for information only, weight of single pup on Day 1. Not included in statistical evaluation105Huntingdon Life SciencesWAG0015

TABLE 25 Sperm analysis - gr oup mean values Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - tr ifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Motile Progressively ------ Cauda epididymis ------ -------- Testis --------

sperm motile sperm Weight Sperm count Total Weight Sperm count Total (%) (%) (g) (millio ns/g) (millio

n) (g) (millio ns/g) (millio

n) Statistical test: Sh Wi Wi Wi Wi Sh Wi Wi 1 Mean 94 59 0.240 1174 281 1.80 199 359 SD 3 15 0.026 160 62 0.13 37 71 n 10 10 10 9 9 10 10 10 2 Mean 95 48 0.237 1145 269 1.84 195 362 SD 4 19 0.016 187 37 0.10 65 135 n 10 10 10 10 10 10 10 10 3 Mean 92 44 0.219 957 223 1.66 148 270 SD 11 21 0.046 396 94 0.49 75 157 n 9 9 10 10 10 10 10 10 4 Mean 92 43* 0.199** 1003 197** 1.74 179 312 SD 4 11 0.027 209 37 0.11 43 85 n 10 10 10 10 10 10 10 10106Huntingdon Life SciencesWAG0015

TABLE 26 Sperm motion data - gr oup mean values Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3 ,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group VAP VSL VCL ALH BCF STR LIN Elongation Area Rapid Medium Slow Static

(um/s) (um/s) (um/s) (um) (Hz) (%) (%) (%) (um sq) (%) (%) (%) (%) Statistical test: Wi Wi Wi Wi Wi Wi Wi Wi Wi Wi Wi Wi Sh 1 Mean 138 102 299 22 25 74 35 24 487 67 3 24 6 SD 8 9 24 2 2 4 2 5 95 15 2 15 3 n 10 10 10 10 10 10 10 10 10 10 10 10 10 2 Mean 126* 92 265** 22 27 73 36 25 506 54 5* 36 5 SD 13 10 22 1 2 4 2 4 124 19 2 19 4 n 10 10 10 10 10 10 10 10 10 10 10 10 10 3 Mean 119** 85* 240** 21 26 71 37 25 445 52 5* 36 8 SD 14 13 29 2 3 3 2 2 87 25 2 22 11 n 9 9 9 9 9 9 9 9 9 9 9 9 9 4 Mean 125** 90** 260** 22 28** 72 37 26 525 51 7** 35 8 SD 13 13 31 1 1 4 2 4 155 13 2 10 4 n 10 10 10 10 10 10 10 10 10 10 10 10 10107Huntingdon Life SciencesWAG0015

TABLE 27 Sperm mo

rphology - gr oup mean values Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Number Total number Normal Total ab normal Decapitate Head ab normal Detached/broken Other of of sperm neck/midpiece/tail abnormality animals examined number % number % number % number % number % number % Statistical test: Sh lWi Sh Sh Sh Wi 1 10 2000 Mean 197 98.6 3 1.5 1.4 0.7 1.0 0.5 0.2 0.1 0.4 0.2 SD 2 0.9 2 0.9 2.1 1.0 0.8 0.4 0.6 0.3 0.7 0.3 2 10 2013 Mean 194 96.6 7 3.4** 3.0 1.5 3.0 1.5 0.6 0.3 0.4 0.2 SD 5 2.2 5 2.2 2.9 1.5 3.6 1.8 0.7 0.3 0.5 0.3 3 8 1600 Mean 192 95.8* 8 4.2** 4.8 2.4 2.6 1.3 1.3 0.6* 0.1 0.1 SD 4 1.9 4 1.9 4.4 2.2 2.7 1.4 1.2 0.6 0.4 0.2 4 10 2000 Me an 181 90.3** 20 9.8** 10.3 5.2** 6.8 3.4** 2.7 1.4** 1.2 0.6** SD 12 6.1 12 6.1 8.9 4.5 4.8 2.4 2.3 1.2 0.8 0.4108Huntingdon Life SciencesWAG0015

TABLE 28 Request ID: 284533 Organ weights - gr oup mean unadjusted and adjusted values (g) for males - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group /Sex Terminal

Bodyweight Epididymides Pituitary Prostate Seminal Vesicles Testes Un adjusted Means Statistical test: Wi Sh 1M Mean 543 1.337 0.015 1.364 1.667 3.59 SD 32 0.120 0.002 0.165 0.406 0.26 N 10 10 10 10 10 10 2M Mean 483** 1.269 0.011 1.008 1.357 3.68 SD 54 0.048 0.002 0.078 0.320 0.20 N 10 10 10 10 10 10 3M Mean 453** 1.214 0.011 0.937 1.128 3.50 SD 28 0.147 0.002 0.249 0.173 0.48 N 10 10 10 10 10 10 4M Mean 423** 1.164** 0.009 0.625 1.125 3.46 SD 30 0.056 0.002 0.169 0.277 0.23 N 10 10 10 10 10 10 Adjusted Means Statistical test: Wi Wi Wi Wi 1M Mean 0.013 1.297 1.627 3.55 2M Mean 0.011* 1.000** 1.352 3.67 3M Mean 0.011* 0.960** 1.142* 3.52 4M Mean 0.010* 0.677** 1.156* 3.49109Huntingdon Life SciencesWAG0015

TABLE 29 Request ID: 289375 Macropathology - group distribution of findings for males - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group/Sex: 1M 2M 3M 4M Tissue and Finding Number Examined: 10 10 10 10 Prostate Small 0 1 7 10 Lt. epididymis Small 0 0 1 0 Lt. testis Prominent tubules 0 0 1 0 Small 0 0 1 0 Teeth Incisor(s) pale 1 0 6 10 Spleen Adhesions 0 2 3 4 Capsule thickened 0 8 6 9 Skin Hair loss 0 1 2 4 Scab(s) 0 0 1 0110Huntingdon Life SciencesWAG0015

TABLE 29 - continued Request ID: 289377 Macropathology - group distribution of findings for decedent females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group/Sex: 1F 2F 3F 4F Tissue and Finding Number Examined: 0 0 1 2 Teeth Incisor(s) pale 0 0 0 2 Spleen Adhesions 0 0 0 1 Kidneys Pale area(s) 0 0 0 1 Pelvic dilatation 0 0 0 1111Huntingdon Life SciencesWAG0015

TABLE 29 - continued Request ID: 289386 Macropathology - group distribution of findings for females killed after scheduled treatment period - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group/Sex: 1F 2F 3F 4F Tissue and Finding Number Examined: 10 10 9 8 Ovaries Cyst(s) 0 0 1 0 Vagina Abnormal contents 0 0 0 1 Teeth Incisor(s) pale 0 0 3 8 Spleen Adhesions 0 0 1 4 Capsule thickened 0 0 2 3 Enlarged 0 0 0 1 Skin Hair loss 2 2 0 0 Kidneys Irregular surface 0 0 0 1 Pale area(s) 0 0 0 1 Pelvic dilatation 0 1 0 1112Huntingdon Life SciencesWAG0015

TABLE 29 - continued Request ID: 289386 Macropathology - group distribution of findings for females killed after scheduled treatment period - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group/Sex: 1F 2F 3F 4F Tissue and Fin ding Number Examined: 10 10 9 8 Stomach Cyst(s) 0 1 0 0 Liver Pale 0 0 0 1113Huntingdon Life SciencesWAG0015

TABLE 29 - continued Request ID: 289378 Macropathology - group distribution of findings for all females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group/Sex: 1F 2F 3F 4F Tissue and Finding Number Examined: 10 10 10 10 Ovaries Cyst(s) 0 0 1 0 Vagina Abnormal contents 0 0 0 1 Teeth Incisor(s) pale 0 0 3 10 Spleen Adhesions 0 0 1 5 Capsule thickened 0 0 2 3 Enlarged 0 0 0 1 Skin Hair loss 2 2 0 0 Kidneys Irregular surface 0 0 0 1 Pale area(s) 0 0 0 2 Pelvic dilatation 0 1 0 2114Huntingdon Life SciencesWAG0015

TABLE 29 - continued Request ID: 289378 Macropathology - group distribution of findings for all females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group/Sex: 1F 2F 3F 4F Tissue and Finding Number Examined: 10 10 10 10 Stomach Cyst(s) 0 1 0 0 Liver Pale 0 0 0 1115Huntingdon Life SciencesWAG0015

TABLE 30 Request ID: 289389 Histopathology - gr oup distribution of findings for males - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure l evel (ppm) : 0 198 505 2900 Group/Sex: 1M 2M 3M 4M Tissue and Finding Number: 10 10 10 10 Rt. Testis Number Examined: 10 0 0 10 Spermatocytes Degeneration 0 0 0 1 Spleen Number Examined: 0 8 7 9 Adhesions/Inflammation/Fibrosis 0 5 4 6 Capsular Thickening 0 4 6 7 Capsular/Subcapsular Inflammation 0 8 6 7 Prostate Number Examined: 10 10 10 10 Abscessation 0 0 1 1 Acinar Cell Atrophy 1 0 2 0 Interstitial Inflammation 2 2 3 3 Skin Number Examined: 0 0 1 0 Epidermal Hyperplasia 0 0 1 0 Scab(s) 0 0 1 0116Huntingdon Life SciencesWAG0015

TABLE 30 - continued Request ID: 289391 Histopathology - gr oup distribution of findings for decedent females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group/Sex: 1F 2F 3F 4F Tissue and Finding Number: 0 0 1 2 Spleen Number Examined: 0 0 0 1 Adhesions/Inflammation/Fibrosis 0 0 0 1 Capsular Thickening 0 0 0 1 Extramedullary Haemopoiesis 0 0 0 1 Kidneys Number Examined: 0 0 0 1 Cortical Tubular Basophilia 0 0 0 1 Cortical Tubular Dilatation 0 0 0 1 Cortical Tubular Necrosis 0 0 0 1 Pelvic Dilatation 0 0 0 1117Huntingdon Life SciencesWAG0015

TABLE 30 - continued Request ID: 289390 Histopathology - gr oup distribution of findings for females killed after scheduled treatment period - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group/Sex: 1F 2F 3F 4F Tissue and Finding Number: 10 10 9 8 Ovaries Number Examined: 10 10 9 8 Cyst(s) 0 0 1 0 Reduced Size of Corpora Lutea 0 1 0 4 Spleen Number Examined: 0 0 2 6 Adhesions/Inflammation/Fibrosis 0 0 2 4 Capsular Thickening 0 0 2 6 Capsular/Subcapsular Inflammation 0 0 1 4 Extramedullary Haemopoiesis 0 0 1 2 Kidneys Number Examined: 0 1 0 2 Cortical Tubular Basophilia 0 1 0 1 Cortical Tubular Dilatation 0 1 0 2 Pelvic Dilatation 0 1 0 1 Stomach Number Examined: 0 1 0 0 Squamous Cyst - Limiting Ridge 0 1 0 0118Huntingdon Life SciencesWAG0015

TABLE 30 - continued Request ID: 289392 Histopath ology - gr oup distribution of findings for all females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group/Sex: 1F 2F 3F 4F Tissue and Finding Number: 10 10 10 10 Ovaries Number Examined: 10 10 10 10 Cyst(s) 0 0 1 0 Reduced Size of Corpora Lutea 0 1 0 4 Spleen Number Examined: 0 0 2 7 Adhesions/Inflammation/Fibrosis 0 0 2 5 Capsula r Thickening 0 0 2 7 Capsular/Subcapsular Inflammation 0 0 1 4 Extramedullary Haemopoiesis 0 0 1 3 Kidneys Number Examined: 0 1 0 3 Cortical Tubular Basophilia 0 1 0 2 Cortical Tubular Dilatation 0 1 0 3 Cortical Tubular Necrosi s 0 0 0 1 Pelvic Dilatation 0 1 0 2 Stomach Number Examined: 0 1 0 0 Squamous Cyst - Limiting Ridge 0 1 0 0119Huntingdon Life SciencesWAG0015

TABLE 3 1 Primordial ovarian follicle counts - gr oup mean values (F0 generation ) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Follicle /Sex count 1F Mean 16.8

SD 8.94 N 10 2F Mean 17.9

7.09 N 10 3F Mean 12.3

SD 5. 78 N 10 4F Mean 15.4

SD 7.13 N 10120Huntingdon Life SciencesWAG0015

APPENDIX 1 Request ID: 284403 Clinical signs - individual observations for males - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group /Sex Animal Number Death

Code Phase Category Observation Week(s) 3M 222 T Pre - treatment No abnormalities detected Treatment Teeth Abnormal Colour, Pale 8 223 T Pre - treatment No abnormalities detected

Treatment Coat Hair loss, Head 6 - 8 225 T Pre - treatment No abnormalities detected

Treatment Teeth Abnormal Colour, Pale 8 229 T Pre - treatment No abnormalities detected

Treatment Teeth Abnormal Colour, Pale 8 T Term inal sacrifice Only animals with observations are presented121Huntingdon Life SciencesWAG0015

APPENDIX 1 - continued Request ID: 284403 Clinical signs - individual observations for males - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group /Sex Animal Number Death

Code Phase Category Observation Week(s) 4M 231 T Pre - treatment No abnormalities detected Treatment Coat Hair loss, Cervical 8 Hair loss, Head 6 - 8 Teeth Abnormal Colour, Pale 7 - 8 232 T Pre - treatment No abnormalities detected

Treatment Coat Hair loss, Head 7 - 8

Teeth Abnormal Colour, Pale 7 - 8 233 T Pre - treatment No abnormalities detected

Treatment Coat Hair loss, Cervical 8 Hair loss, Head 7 - 8 Teeth Abnormal Colour, Pale 7 - 8 234 T Pre - treatment No abnormalities detected

Treatment Coat Hair loss, Cervical (Left) 8 Hair loss, Head 5 - 8 Teeth Abnormal Colour, Pale 7 - 8 235 T Pre - treatment No abnormalities detected

Treatment Teeth Abnormal Colour, Pale 5 - 8 236 T Pre - treatment No abnormalities detected

Treatment Teeth Abnormal Colour, Pale 6 - 7 T Terminal sacrifice Only animals with observations are presented122Huntingdon Life SciencesWAG0015

APPENDIX 1 - continued Request ID: 284403 Clinical signs - individual observations for males - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group /Sex Animal Number Death

Code Phase Category Observation Week(s) 4M 237 T Pre - treatment No abnormalities detected Treatment Teeth Abnormal Colour, Pale 6 - 7 238 T Pre - treatment No abnormalities detected

Treatment Teeth Abnormal Colour, Pale 5 - 7 239 T Pre - treatment No abnormalities detected

Treatment Coat Hair loss, Head 7

Teeth Abnormal Colour, Pale 5 - 7 240 T Pre - treatment No abnormalities detected

Treatment Teeth Abnormal Colour, Pale 5 - 7 T Terminal sacrifice Only animals with observations are presented123Huntingdon Life SciencesWAG0015

APPENDIX 2 Request ID: 284404 Clinical signs - individual observations for femal es - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group /Sex Animal Number Death

Code Phase Category Observation Day (s) 1F 241 T Pre - treatment No abnormalities detected Treatment No abnormalities detected Gestation No abnormalities detected Lactatio n Coat Hair loss, Forelimbs 9 - 10 245 T Pre - treatment No abnormalities detected

Treatment Behaviour Irritable 12 Vocalisation 12 Gestation No abnormalities detected Lactatio n Coat Hair loss, Forelimbs 5, 9 - 10 247 T Pre - treatment No abnormalities detected

Treatment No abnormalities detected Ges tation No abnormalities detected Lactatio n Staining Abnormal Colour, Brown, Head 10 248 T Pre - treatment No abnormalities detected

Treatment No abnormalities detected Gestation No abnormalities detected Lactatio n Coat Hair loss, Forelimbs 10 Hair loss, Hindlimb (Right) 5, 9 T Terminal sacrifice Only animals with observations are presented124Huntingdon Life SciencesWAG0015

APPENDIX 2 - continued Request ID: 284404 Clinical signs - individual observations for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group /Sex Animal Number Death

Code Phase Category Observation Day (s) 1F 250 T Pre - treatment No abnormalities detected Treatment Coat Hair loss, Head 12, 19 Gestation Coat Hair loss, Forelimbs 14, 20 Hair loss, Head 0, 7, 14, 20 Lactatio n Coat Hair loss, Forelimbs 1, 5, 9 - 10 Hair loss, Head 1, 5, 9 T Terminal sacrifice Only animals with observations are presented125Huntingdon Life SciencesWAG0015

APPENDIX 2 - continued Request ID: 284404 Clinical signs - individual observations for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group /Sex Animal Number Death

Code Phase Category Observation Day (s) 2F 251 T Pre - treatment No abnormalities detected Treatment No abnormalities detected Gestation No abnormalities detected Lactatio n Staining Abnormal Colour, Brown, Head 9 - 10 252 T Pre - treatment No abnormalities detected

Treatment No abnormalities detected Gestation No abnormalities det ected Lactatio n Coat Hair loss, Forelimbs 10 253 T Pre - treatment No abnormalities detected

Treatment No abnormalities detected Gestation No abnormalities detected Lactatio n Coat Hair loss, Forelimbs 9 - 10 Hair loss, Ventral Body Surface 9 - 10 255 T Pre - treatment No abnormalities detected

Treatment No abnormalities detected Gestation Coat Hair loss, Forelimb (Right) 20 Lactatio n Coat Hair loss, Forelimb (Left) 9 - 10 Hair loss, Forelimb (Righ t) 5, 9 - 10 Hair loss, Hindlimb (Right) 10 Hair loss, Ventral Body Surface 5, 9 - 10 T Terminal sacrifice Only animals with observations are presented126Huntingdon Life SciencesWAG0015

APPENDIX 2 - continued Request ID: 284404 Clinical signs - individual observation s for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group /Sex Animal Number Death

Code Phase Category Observation Day (s) 2F 257 NP T Pre - treatment No abnormalities detected Treatment No abnormalities detected Gestation Reason for Dispatch Failed to litter (non - pr egnant) 25

Comments: failed to litter 25 258 T Pre - treatme nt No abnormalities detected Treatment No abnormalities detected Gestation No abnormalities detected Lactatio n Staining Abnormal Colour, Brown, Head 9 - 10 259 T Pre - treatment No abnormalities detected

Treatment No abnormalities detected Gestation No abnormalities detected Lactatio n Coat Hair loss, Forelimbs 5, 9 - 10 Hair loss, Ventral Body Surface 10 260 T Pre - treatment No abnormalities detected

Treatment No abnormalities detected Gestation Staining Abnormal Colour, Brown, Head 14, 20 Lactatio n No abnormalities detected T Terminal sacrifice NP Not pregnant Only animals with observations are presented127Huntingdon Life SciencesWAG0015

APPENDIX 2 - continued Request ID: 284404 Clinical signs - individu al observations for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group /Sex Animal Number Death

Code Phase Category Observation Day (s) 3F 261 T Pre - treatment No abnormalities detected Treatment No abnormalities detected Gestation Teeth Abnormal Colour, Pale 14, 20 Lactatio n Teeth Abnormal Colour, Pale 1, 5, 9 - 10 262 W Pr e - treatment No abnormalities detected

Treatment No abnormalities detected Gestation No abnormalities detected Lactatio n Staining Abnormal Colour, Head 1

Other No evidence of lactating 1 Reason for Dispatch Total litter loss post partum/general poor condition of dam 1 264 T Pre - treatment Behaviour Aggressive 5 Vocalisation 5 Treatment Behaviour Aggressive 5, 12, 19 Vocalisation 5, 12, 19 Gestation Behaviour Irritable 7 Lactatio n No abnormalities detected 265 TLL Pre - treatment No abnormalities detected

Treatment No abnormalities detected Gestation No abnormalities detected Lactatio n Reason for Dispatch Total litter loss post partum 1

Comments: Total Litter Loss 1 T Terminal sacrifice W Killed for welfare reasons TLL Total litter loss Only animals with observations are presented128Huntingdon Life SciencesWAG0015

APPENDIX 2 - continued Request ID: 284404 Clinical signs - individual observations for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group /Sex Animal Number Death

Code Phase Category Observation Day (s) 3F 266 TLL Pre - treatment No abnormalities detected Treatment No abnormalities detected Gestation Coat Hair loss, Forelimbs 25 Hair loss, Head 0, 7, 14, 20 Hair loss, Ventral Body Surface 20, 25 Teeth Abnormal Colour, Pale 14, 20, 25 Oth er Showing no interest in offspring Pre Day 1 lactatio n Reason for Dispatch Total litter loss (prior to Day 1 lactation) A 267 TLL Pre - treatment No abnormalities detected

Treatment No abnormalities detected Gestation Teeth Abnormal Colour, Pale 14, 20 Lactatio n Reason for Dispatch General poor clinical condition 1

Teeth Abnormal Colour, Pale 1 Other No evidence of lactating 1 Reason for despatch Total litter Loss 1 268 T Pre - treatment No abnormaliti es detected

Treatment No abnormalities detected Gestation Staining Abnormal Colour, Brown, Head 14, 20 Lactatio n Staining Abnormal Colour, Brown, Head 10 TLL Total litter loss A Total litter loss prior to Day 1 lactation and assignment to the lactation phase on the data capture system T Terminal sacrifice Only animals with observations are presented129Huntingdon Life SciencesWAG0015

APPENDIX 2 - continued Request ID: 284404 Clinical signs - individual observations for females - F0 generatio n Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group /Sex Animal Number Death

Code Phase Category Observation Day (s) 3F 269 T Pre - treatment No abnormalities detected Treatment No abnormalities detected Gestation Staining Abnormal Colour, Brown, Head 14, 20 Lactatio n Staining Abnormal Colour, Brown, Head 5, 9 - 10 270 TLL Pre - treatment No abnormalities detected

Treatment No abnormalities detected Gestation No abnormalities detected Lactatio n No abnormalities detected

Reason for despatch Total litter loss 2 TLL Total litter loss T Terminal sac

rifice Only animals with observations are presented130Huntingdon Life SciencesWAG0015

APPENDIX 2 - continued Request ID: 284404 Clinical signs - individual observations for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group /Sex Animal Number Death

Code Phase Category Observation Day (s) 4F 271 TR T Pre - treatment No abnormalities detected Treatment Build Co nformation Thin 12, 19 Coat Piloerection 8 - 10 Posture Hunched 8 - 10 Staining Abnormal Colour, Brown, Muzzle 11 - 12, 19 Gestation Behaviour Un deractive 13 - 14

Build Co nformation Thin 0 Coat Piloerection 13 - 15, 20, 25 Eyelids Partially Clo sed 13 - 14 Muscle Reaction Reduced Body Tone, Whole Body 13 - 14 Staining Abnormal Colour, Brown, Head 15, 20, 25

Abnormal Colour, Brown, Muzzle 0, 7, 13 - 14 Reason for Dispatch Failed to litter 25 272 FTM T Pre - treatment No abn ormalities detected

Treatment Muscle Reaction Reduced Body Tone 17, 19, 26, 33, 40, 47, 54

Posture Hunched 17, 19, 26, 33 Staining Abnormal Colour, Eye (Left) 40, 47, 54

Abnormal Colour, Eye (Right) 40, 47, 54 Abnormal Colour, Re d, Muzzle 16 - 17, 19, 26, 33, 40, 47 Teeth Abnormal Colour, Pale 33, 40, 47, 54 Reason for despatch Failed to mate 54 T Terminal sacrifice TR Total resorption FTM Failed to mate Only animals with observations are presented131Huntingdon Life SciencesWAG0015

APPENDIX 2 - continued Request ID: 284404 Clinical signs - individual observations for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (pp m) : 0 198 505 2900 Group /Sex Animal Number Death

Code Phase Category Observation Day (s) 4F 273 TLL Pre - treatment Behaviour Aggressive 5 Vocalisation 5 Treatment Behaviour Aggressive 5, 8 - 10 Vocalisation 5, 8 - 10 Coat Piloer ection 8 - 10 Posture Hunched 8 - 10 Gestation Teeth Abnormal Colour, Pale 20 Lactatio n Reason for Dispatch General poor clinical condition 2

Teeth Abnormal Colour, Pale 1 Reason for despatch Total litter loss 2 274 TR T Pre - tr eatment No abnormalities detected

Treatment Muscle Reaction Reduced Body Tone 16 - 17

Staining Abnormal Colour, Brown, Head 16 - 17 Gestation Muscle Reaction Reduced Body Tone 0

Reason for Dispatch General poor clinical condition 25 St aining Abnormal Colour, Brown, Head 0, 7, 14, 20, 25 Teeth Abnormal Colour, Pale 14, 20, 25 Reason for despatch Failed to litter 25 275 FTM T Pre - treatment No abnormalities detected

Treatment Staining Abnormal Colour, Brown, Head 54

Teeth Abnormal Colour, Pale 33, 40, 47, 54 Reason for despatch Failed to mate 54 TLL Total litter loss T Terminal sacrifice TR Total resorption FTM Failed to mate Only animals with observations are presented132Huntingdon Life SciencesWAG0015

APPENDIX 2 - continued Request ID: 284404 Clinical signs - individual observations for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group /Sex Animal Number Death

Code Phase Category Observation Day (s) 4F 276 W Pre - treatment No abnormalities detected Treatment No abnormalities detected Gestation Behaviour Un deractive 24

Body temperatur e Reduced 24 Coat Piloerection 24 Posture Hunched 24 Staining Abnormal Colour, Brown, Head 7, 14, 20, 24 Teeth Abnormal Colour, Pale 14, 20, 24 Reason for Dispatch General poor clinical condition 24 277 FTM T Pre - treatmen t No abnormalities detected

Treatment Coat Piloerection 13, 16 - 17, 19, 26

Posture Hunched 13, 16 - 17, 19, 26 Staining Abnormal Colour, Brown, Head 12 - 13, 16 - 17, 19, 26, 33, 40, 47, 54 Teeth Abnormal Colour, Pale 33, 40, 47, 54 Reas on for Dispatch Failed to mate 54 278 TR T Pre - treatment No abnormalities detected

Treatment No abnormalities detected Gestation Staining Abnormal Colour, Brown, Muzzle 0, 7, 14, 20, 25

Teeth Abnormal Colour, Pale 14, 20, 25 Reason for Dispatch Failed to litter 25 W Killed for welfare reasons T Terminal sacrifice FTM Failed to mate TR Total resorption Only animals with observations are presented133Huntingdon Life SciencesWAG0015

APPENDIX 2 - continued Request ID: 284404 Clini cal signs - individual observations for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group /Sex Anima l Number Death

Code Phase Category Observation Day (s) 4F 279 W Pre - treatment Behaviour Vocalisation 5 Treatment Behaviour Vocalisation 5, 12, 19, 26

Staining Abnormal Colour, Brown, Head 12, 19, 26 Gestation Coat Piloerection 24

Staining Abnormal Colour, Brown, Head 0, 7, 14, 20

Abnormal Colour, Red, Perigenital 24 Teeth Abnormal Colour, Pale 7, 14, 20, 24 Other Appeared distressed and attempting parturition 24 Reason for Dispatch General poor clinical conditi on 24 280 NP T Pre - treatment No abnormalities detected

Treatment Staining Abnormal Colour, Brown, Head 12 Gestation Staining Abnormal Colour, Brown, Head 0, 7, 14, 20, 25

Teeth Abnormal Colour, Pale 25 Reason for Dispatch Fail ed To Litter 25 W Killed for welfare reasons T Terminal sacrifice NP Not pregnant Only animals with observations are presented134Huntingdon Life SciencesWAG0015

APPENDIX 3 Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 2M 211 Behaviour

Underactive 1 1, 4 Unresponsive 1 1 Coat Piloerection 1, 5 1 - 2, 4 - 5 Eyel ids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2 212 Behaviour Underactive 1 1, 4 Unresponsive 1 1 Coat Piloerection 1 1 - 2, 4 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2135Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) Durin g dosing

(~4 hours) On return to the

home cage At the end of the

working day 2M 213 Behaviour

Underactive 1 1, 4 Unresponsive 1 1 Coat Piloerection 1 1 - 2, 4 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2 214 Behaviour Underactive 1 1, 4 Unresponsive 1 1 Coat Piloerection 1 1 - 2, 4 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2136Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

hom e cage At the end of the

working day 2M 215 Behaviour

Underactive 1 1, 4 Unresponsive 1 1 Coat Piloerection 1 1 - 2, 4 Eyelids (Left) Partially Closed 2 Eyelids (Right) Pa rtially Closed 2 216 Behaviour Underactive 1 1, 4 Unresponsive 1 1 Coat Piloerection 1 1 - 2, 4 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2137Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 2M 217 Behaviour

Underactive 1 1, 4 Unresponsive 1 1 Coat Piloerection 1 1 - 2, 4 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2 218 Behaviour Underactive 1 1, 4 Unresponsive 1 1 Coat Piloerection 1 1 - 2, 4 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2138Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Re quest ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 2M 219 Behaviour

Underactive 1 1, 4 Unresponsive 1 1 Coat Piloerection 1 1 - 2, 4 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2 220 Behaviour Underactive 1 1, 4 Unresponsive 1 1 Coat Piloerection 1 1 - 2, 4 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2139Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 50 5 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 3M 221 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1, 3 - 4 1, 3 - 4 Br eathing Shallow 2 Slow 5 4 - 5 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Partially Closed 1 - 2, 5 1 - 2, 5 Eyelids (Right) Partially Closed 1 - 2, 5 1 - 2, 5 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1140Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Grou p : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 3M 222 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1, 3 - 4 1, 3 - 4 Br eathing Shallow 2 Slow 5 4 - 5 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Partially Closed 1 - 2, 5 1 - 2, 5 Eyelids (Right) Partially Closed 1 - 2, 5 1 - 2, 5 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1141Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 3M 223 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1, 3 - 4 1, 3 - 4 Br eathing Shallow 2 Slow 5 4 - 5 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Partially Closed 1 - 2, 5 1 - 2, 5 Eye lids (Right) Partially Closed 1 - 2, 5 1 - 2, 5 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1142Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 3M 224 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1, 3 - 4 1, 3 - 4 Br eathing Shallow 2 Slow 5 4 - 5 Coat Piloerection 1 - 7 1 - 7 1 Eyelids (Left) Partially Closed 1 - 2, 5 1 - 2, 5 Eyelids (Right) Partially Closed 1 - 2, 5 1 - 2, 5 Eye (Left) Lachrymatio

n 1 Eye (Right) Lachrymation 1143Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On retu

rn to the home cage At the end of the

working day 3M 225 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1, 3 - 4 1, 3 - 4 Br eathing Shallow 2 Slow 5 4 - 5 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Partially Closed 1 - 2, 5 1 - 2, 5 Eyelids (Right) Partially Closed 1 - 2, 5 1 - 2, 5 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1144Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Requ est ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropr opene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 3M 226 Behaviour

Undera ctive 1 - 7 1 - 7 Unresponsive 1, 3 - 4 1, 3 - 4 Br eathing Shallow 2 Slow 5 4 - 5 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Partially Closed 1 - 2, 5 1 - 2, 5 Eyelids (Right) Partially Closed 1 - 2, 5 1 - 2, 5 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1145Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females d ur ing treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 3M 227 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1, 3 - 4 1, 3 - 4 Br eathing Shallow 2 Slow 5 4 - 5 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Partially Closed 1 - 2, 5 1 - 2, 5 Eyelids (Right) Partially Closed 1 - 2, 5 1 - 2, 5 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1146Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the en d of the

working day 3M 228 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1, 3 - 4 1, 3 - 4 Br eathing Shallow 2 Slow 5 4 - 5 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Pa rtially Closed 1 - 2, 5 1 - 2, 5 Eyelids (Right) Partially Closed 1 - 2, 5 1 - 2, 5 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1147Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associ ated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 3M 229 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1, 3 - 4 1, 3 - 4 Br eathing Shallow 2 Slow 5 4 - 5 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Partially Closed 1 - 2, 5 1 - 2, 5 Eyelids (Right) Partially Closed 1 - 2, 5 1 - 2, 5 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1148Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generati on Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hour s) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 3M 230 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1, 3 - 4 1, 3 - 4 Br eathing Shallow 2 Slow 5 4 - 5 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Partially Closed 1 - 2, 5 1 - 2, 5 Eyelids (Right) Partially Closed 1 - 2, 5 1 - 2, 5 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1149Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 4M 231 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1 - 7 1 - 7 Br eathing Shallow 1 1 - 2 Slow 4 - 6 4 - 6 Coat Piloerection 1 - 7 1 - 7 1 Eyelids (Left) Partially Closed 1 - 7 1 - 7 Eyelids (Right) Partially Closed 1 - 7 1 - 7150Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On retu

rn to the home cage At the end of the

working day 4M 232 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1 - 7 1 - 7 Br eathing Shallow 1 1 - 2 Slow 4 - 6 4 - 6 Coat Piloerection 1 - 7 1 - 7 E yelids (Left) Partially Closed 1 - 7 1 - 7 Eyelids (Right) Partially Closed 1 - 7 1 - 7151Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during tre atment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 4M 233 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1 - 7 1 - 7 Br eathing Shallow 1 1 - 2 Slow 4 - 6 4 - 6 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Partially Closed 1 - 7 1 - 7 Eyelids (Right) Partially Closed 1 - 7 1 - 7152Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 4M 234 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1 - 7 1 - 7 Br eathing Shallow 1 1 - 2 Slow 4 - 6 4 - 6 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Partially Closed 1 - 7 1 - 7 Eyelids (Right) Partially Closed 1 - 7 1 - 7153Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

t rifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 4M 235 Behaviour

Underactive 1 - 7 1 - 7 1 Unresponsive 1 - 7 1 - 7 Br eathing Shallow 1 1 - 2 Slow 4 - 6 4 - 6 Coat Piloerection 1 - 7 1 - 7 1 Eyelids (Left) Partially Closed 1 - 7 1 - 7 Eyelids (Right) Partially Closed 1 - 7 1 - 7154Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trif luoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 4M 236 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1 - 7 1 - 7 Br eathing Shallow 1 1 - 2 Slow 4 - 6 4 - 6 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Partially Closed 1 - 7 1 - 7 Eyelids (Right) Partially Closed 1 - 7 1 - 7155Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 4M 237 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1 - 7 1 - 7 Br eathing Shallow 1 1 - 2 Slow 4 - 6 4 - 6 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Partially Closed 1 - 7 1 - 7 Eyelids (Right) Partially Closed 1 - 7 1 - 7156Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations f ormales and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 4M 238 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1 - 7 1 - 7 Br eathing Shallow 1 1 - 2 Slow 4 - 6 4 - 6 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Partially Closed 1 - 7 1 - 7 Eyelids (Right) Partially Closed 1 - 7 1 - 7157Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 4M 239 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1 - 7 1 - 7 Br eathing Shallow 1 1 - 2 Slow 4 - 6 4 - 6 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Partially Closed 1 - 7 1 - 7 Eyelids (Right) Partially Closed 1 - 7 1 - 7158Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

t rifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 4M 240 Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1 - 7 1 - 7 Br eathing Shallow 1 1 - 2 Slow 4 - 6 4 - 6 Coat Piloerection 1 - 7 1 - 7 1 Eyelids (Left) Partially Closed 1 - 7 1 - 7 Eyelids (Right) Partially Closed 1 - 7 1 - 7159Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compoun d : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 2F 251 Behaviour

Underactive 1 1 Unresponsive 1 1 Coat Piloerection 1 1 - 2 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2 252 Behaviour Underactive 1 1 Unresponsive 1 1 Coat Piloerection 1 1 - 2 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Close d 2160Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3 ,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 2F 253 Behaviour

Underactive 1 1 Unresponsive 1 1 Coat Piloerection 1 1 - 2 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2 254 Behaviour Underactive 1 1 Unresponsive 1 1 Coat Piloerection 1 1 - 2 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2161Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request I D: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 2F 255 Behaviour

Underactive 1 1, 4 Unresponsive 1 1 Coat Piloerection 1 1 - 2, 4 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2 256 Behaviour Underactive 1 1 Unr esponsive 1 1 Coat Piloerection 1 1 - 2 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2162Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 2F 257 Behaviour

Underactive 1 1 Unresponsive 1 1 Coat Piloerection 1 1 - 2 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2 258 Behaviour Underactive 1 1 Unresponsive 1 1 Coat Piloerection 1 1 - 2 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2163Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during tr eatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 2F 259 Behaviour

Underactive 1 1 Unresponsive 1 1 Coat Piloerection 1 1 - 2 Eyelids (Left) Partially Closed 2 Eyelids (Right) Partially Closed 2 260 Behaviour Underactive 1 1 Unresponsive 1 1 Coat Piloerection 1 1 - 2 Eyelids (Left) Partially Close d 2 Eyelids (Right) Partially Closed 2164Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) O n return to the

home cage At the end of the

working day 3F 261 Behaviour

Underactive 1 - 3 1 - 3 Unresponsive 1 1, 3 Br eathing Shallow 2 Coat Piloerection 1 - 3 1 - 3 Eyelids (Left) Partially Closed 1 - 2 1 - 2 Eyelids (Right) Partially Closed 1 - 2 1 - 2 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1165Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 19 8 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 3F 262 Behaviour

Underactive 1 - 3 1 - 3 Unresponsi ve 1 1, 3 Br eathing Shallow 2 Coat Piloerection 1 - 3 1 - 3 Eyelids (Left) Partially Closed 1 - 2 1 - 2 Eyelids (Right) Partially Closed 1 - 2 1 - 2 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1166Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to th e

home cage At the end of the

working day 3F 263 Behaviour

Underactive 1 - 3 1 - 3 Unresponsive 1 1, 3 Br eathing Shallow 2 Coat Piloerection 1 - 3 1 - 3 Eyelids (Left) Partially Closed 1 - 2 1 - 2 Eyelids (Right) Partially Closed 1 - 2 1 - 2 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1167Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 3F 264 Behaviour

Underactive 1 - 3 1 - 3 Unresponsive 1 1, 3 Br eathing Shallow 2 Coat Piloerection 1 - 3 1 - 3 Eyelids (Left) Partially Closed 1 - 2 1 - 2 Eyelids (Right) Partially Closed 1 - 2 1 - 2 Eye (Left) Lachrymation 1 Eye (Righ t) Lachrymation 1168Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 3F 265 Behaviour

Underactive 1 - 2 1 - 2 Unresponsive 1 1 Br eathing Shallow 2 Coat Piloerection 1 - 2 1 - 2 Eyelids (Left) Partially Closed 1 - 2 1 - 2 Eyelids (Right) Partially Closed 1 - 2 1 - 2 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1169Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Grou p /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 3F 266 Behaviour

Underactive 1 - 3 1 - 3 Unresponsive 1, 3 1, 3 Br eathing Shallow 2 Coat Piloerection 1 - 3 1 - 3 Eyelids (Left) Partially Closed 1 - 2 1 - 2 Eyelids (Right) Partially Closed 1 - 2 1 - 2 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1170Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of th e

working day 3F 267 Behaviour

Underactive 1 - 3 1 - 3 Unresponsive 1 1, 3 Br eathing Shallow 2 Coat Piloerection 1 - 3 1 - 3 Eyelids (Left) Partially Closed 1 - 2 1 - 2 Eyelid s (Right) Partially Closed 1 - 2 1 - 2 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1171Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 3F 268 Behaviour

Underactive 1 - 3 1 - 3 Unresponsive 1 1, 3 Br eathing Shallow 2 Coat Piloerection 1 - 3 1 - 3 Eyelids (Left) Partially Closed 1 - 2 1 - 2 Eyelids (Right) Partially Closed 1 - 2 1 - 2 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1172Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3 ,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 3F 269 Behaviour

Underactive 1 - 3 1 - 3 Unresponsive 1 1, 3 Br eathing Shallow 2 Coat Piloerection 1 - 3 1 - 3 1 Eyelids (Left) Partially Closed 1 - 2 1 - 2 Eyelids (Right) Partially Closed 1 - 2 1 - 2 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1173Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and fem ales during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 3F 270 Behaviour

Underactive 1 - 3 1 - 3 Unresponsive 1 1, 3 Br eathing Shallow 2 Coat Piloerection 1 - 3 1 - 3 Eyelids (Left) Partially Closed 1 - 2 1 - 2 Eyelids (Right) Partially Closed 1 - 2 1 - 2 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1174Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluorop ropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 4F 271 Behaviour

Underactive 1 - 3 1 - 3 Unresponsive 1 - 3 1 - 3 Br eathing Shallow 1 1 - 2 Coat Piloerection 1 - 3 1 - 3 Eyelids (Left) Partially Closed 1 - 3 1 - 3 Eyelids (Right) P artially Closed 1 - 3 1 - 3 Posture Hun ched 2 2 2 2 Staining Other Colour (see comment) 2 Comments: brown staining head 2175Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 1 98 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 4F 272 FTM Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1 - 7 1 - 7 Br eathing Shallow 1 1 - 2 Slow 4 - 6 4 - 6 Coat Piloerection 1 - 7 1 - 7 1 Eyelids (Left) Partially Closed 1 - 7 1 - 7 Eyelids (Right) Partially Closed 1 - 7 1 - 7 FTM Failed to mate176Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 4F 273 Behaviour

Underactive 1 - 2 1 - 2 Unresponsive 1 - 2 1 - 2 Br eathing Shallow 1 1 - 2 Coat Piloerection 1 - 2 1 - 2 Eyelids (Left) Partially Closed 1 - 2 1 - 2 Eyelids (Righ t) Partially Closed 1 - 2 1 - 2 Posture Hun ched 2 2 2 2 2 Staining Other Colour (see comment) 2 2 Comments: brown staining head 2 2177Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposur e level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 4F 274 Behaviour

Underactive 1 - 2 1 - 2 Unresponsive 1 - 2 1 - 2 Br eathing Shallow 1 1 - 2 Coat Piloerection 1 - 2 1 - 2 Eyelids (Left) Partially Closed 1 - 2 1 - 2 Eyelids (Right) Partially Closed 1 - 2 1 - 2178Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 4F 275 FTM Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1 - 7 1 - 7 Br eathing Shallow 1 1 - 2 Slow 4 - 6 4 - 6 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Partially Closed 1 - 7 1 - 7 Eyelids (Right) Partially Closed 1 - 7 1 - 7 Posture Hun ched 2 Staining Other Colour (see comment) 2 2 2 2 Comments: Brown staining head 2 2 2 2 FTM Failed to mate179Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 4F 276 Behaviour

Un deractive 1 - 3 1 - 3 Unresponsive 1 - 3 1 - 3 Br eathing Shallow 1 1 - 2 Coat Piloerection 1 - 3 1 - 3 Eyelids (Left) Partially Closed 1 - 3 1 - 3 Eyelids (Right) Partially Closed 1 - 3 1 - 3 277 FTM Behaviour

Underactive 1 - 7 1 - 7 Unresponsive 1 - 7 1 - 7 Br eathing Shallow 1 1 - 2 Slow 4 - 6 4 - 6 Coat Piloerection 1 - 7 1 - 7 Eyelids (Left) Partially Closed 1 - 7 1 - 7 Eyelids (Right) Partially Closed 1 - 7 1 - 7 FTM Failed to mate180Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) Durin g dosing

(~4 hours) On return to the

home cage At the end of the

working day 4F 278 Behaviour

Underactive 1 - 3 1 - 3 Unresponsive 1 - 3 1 - 3 Br eathing Shallow 1 1 - 2 Coat Piloerection 1 - 3 1 - 3 Eyelids (Left) Partially Closed 1 - 2 1 - 3 Eyelids (Right) Partially Closed 1 - 2 1 - 3 279 Behaviour Underactive 1 - 4 1 - 4 Unresponsive 1 - 4 1 - 4 Br eathing Shallow 1 1 - 2 Slow 4 4 Coat Piloerection 1 - 3 1 - 3 Eyelids (Left) Partially Closed 1 - 4 1 - 4 Eyelids (Right) Partially Closed 1 - 4 1 - 4181Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284715 Signs associated with dosing - individual observations for males and females during treatment - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 290 0 Weeks Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) During dosing

(~4 hours) On return to the

home cage At the end of the

working day 4F 280 Behaviour

Underactive 1 - 3 1 - 3 Unresponsive 1 - 3 1 - 3 Br eathing Shallow 1 1 - 2 Coat Piloerection 1 - 3 1 - 3 Eyelids (Left) Partially Closed 1 - 2 1 - 3 Eyelids (Right) Partially Closed 1 - 2 1 - 3182Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284712 Signs associated with dosing - individual observations for females after mating - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Days Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) Additional

observation During dosing

(~4 hours) 2F 251 Behaviour

Underactive 7 Coat Piloerection 7, 19 252 Behaviour Underactive 7, 14 0, 7, 14 Coat Piloerection 7, 19 0, 7, 19 253 Coat Piloerection 7 7 254 Coat Piloerection 7 7 255 Behaviour Underactive 14 0, 14 Coat Piloerection 14 0, 14 256 Coat Piloerection 7 7183Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284712 Signs associated with dosing - individual observations for females after mating - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Days Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) Additional

observation During dosing

(~4 hours) 2F 257 NP Coat

Piloerection 7 7 258 Behaviour Underactive 7, 14 0, 7, 14 Coat Piloerection 7, 19 0, 7, 19 259 Behaviour Underactive 7 Coat Piloerection 7, 19 260 Behaviour Underactive 7 Coat Piloerection 7, 19 NP Not pregnant184Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284712 Signs associated with dosing - individual observations for females after mating - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Days Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) Additional

observation During dosing

(~4 hours) 3F 261 Behaviour

Underactive 7, 14, 19 7, 14, 19 Unresponsive 7 7 Br eat hing Slow 7, 19 Coat Piloerection 14 14 Eyelids (Left) Partially Closed 19 Eyelids (Right) Partially Closed 19 262 Behaviour Underactive 0, 7, 14, 19 0, 7, 14, 19 Unresponsive 7 Br eathing Slow 0, 14, 19 0, 14, 19 Coat Piloerection 7, 14 7, 14, 19 Eyelids (Left) Partially Closed 0, 7, 19 0, 7, 14, 19 Eyelids (Right) Partially Closed 0, 7, 19 0, 7, 14, 19185Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284712 Signs associated with dosing - individual observations for females after mating - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - brom o - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Days Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) Additional

observation During dosing

(~4 hours) 3F 263 U TR Behaviour

Underactive 0, 7, 19 0, 7, 14, 19 Unresponsive 0 Br eathing Slow 19 0, 7, 14, 19 Coat Piloerection 19 19 Eyelids (Left) Partially Closed 0, 7, 14 Eyelids (Right) Partially Closed 0, 7, 14 264 Behaviour

Underactive 0, 7, 14, 19 0, 7, 14, 19 Unresponsive 7 Br eathing Slow 0, 14, 19 0, 14, 19 Coat Piloerection 7, 14 7, 14, 19 Eyelids (L eft) Partially Closed 0, 7, 19 0, 7, 14, 19 Eyelids (Right) Partially Closed 0, 7, 19 0, 7, 14, 19 U Un ilateral implantation TR Total resorption186Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284712 Signs associated with dosing - individual observations for females after mating - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 D ays Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) Additional

observation During dosing

(~4 hours) 3F 265 Behaviour

Underactive 0, 7, 14, 19 0, 7, 14, 19 Br eathing Shallow 0 Slow 7, 19 7, 19 Coat Piloerection 0, 14, 19 0, 14, 19 Eyelids (Left) Partially Closed 7, 19 7, 19 Eyelids (Right) Partially Closed 7, 19 7, 19 266 Behaviour Underactive 0, 7, 14, 1 9 0, 7, 14, 19 Unresponsive 19 Br eathing Slow 0 0, 14, 19 Coat Piloerection 7 7 Eyelids (Left) Partially Closed 0 0, 14, 19 Eyelids (Right) Partially Closed 0 0, 14, 19187Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284712 Signs associated with dosing - individual observations for females after mating - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Days Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) Additional

observation During dosing

(~4 hours) 3F 267 Behaviour

Underactive 7, 1 4, 19 7, 14, 19 Unresponsive 7 7 Br eathing Slow 7, 19 Coat Piloerection 14 14 Eyelids (Left) Partially Closed 19 Eyelids (Right) Partially Closed 19 268 Behaviour Underactive 7, 14, 19 7, 14, 19 Unresponsive 7 7 Br eathing Slow 7, 19 Coat Piloerection 14 14 Eyelids (Left) Partially Closed 19 Eyelids (Right) Partially Closed 19188Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284712 Signs associated with dosing - individual observations for females after mating - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

tri fluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Days Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) Additional

observation During dosing

(~4 hours) 3F 269 Behaviour

U nderactive 0, 7, 19 0, 7, 14, 19 Unresponsive 0 Br eathing Slow 19 0, 7, 14, 19 Coat Piloerection 19 19 Eyelids (Left) Partially Closed 0, 7, 14 Eyelids (Right) Partially Cl osed 0, 7, 14 270 Behaviour Underactive 7, 14, 19 7, 14, 19 Unresponsive 7 7 Br eathing Slow 7, 19 Coat Piloerection 14 14 Eyelids (Left) Partially Closed 19 Eyelids (Right) Partially Closed 19189Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284712 Signs associated with dosing - individual observations for females after mating - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3, 3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Days Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) Additional

observation During dosing

(~4 hours) 4F 271 TR Behaviour

Underactive 14 0, 7, 14, 19 0, 7, 14, 19 Unresponsive 0, 19 0, 19 Br eathing Slow 14, 19 7, 14, 19 Coat Piloerection 14 0, 7, 19 0, 7, 19 Eyelids (Left) Pa rtially Closed 14 0, 14, 19 0, 7, 14, 19 Eyelids (Right) Partially Closed 14 0, 14, 19 0, 7, 14, 19 Muscle Reaction Reduced Body Tone 14 TR Total resorption190Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284712 Signs associated with dosing - individual observations for females after mating - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Days Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) Additional

observation During dosing

(~4 hours) 4F 273 Behaviour

Underactive 0, 7, 14, 19 0, 7, 14, 19 Unresponsive 0, 14 0, 14 Br eathing Slow 7, 19 0, 7, 19 Coat Piloerection 0, 7, 14 0, 7, 14 Eyelids (Left) Partially Closed 0, 7, 14, 19 0, 7, 14, 19 Eyelids (Right) Partially Closed 0, 7, 14, 19 0, 7, 14, 19 274 TR Behaviour

Underactive 0, 7, 19 0 7, 14, 19 Unresponsive 0, 14 0 14 Coat Piloerection 0, 7, 19 0 7, 19 Eyelids (Left) Partially Closed 0, 7 0 7 Eyelids (Right) Partially Closed 0, 7 0 7 Posture Hun ched 14 14 TR Total resorption191Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284712 Signs associated with dosing - individual observations for females after mating - F0 generation Group : 1 2 3 4 C ompound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Days Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) Additional

observation During dosing

(~4 hours) 4F 276 W Behaviour

Underactive 0, 7, 14, 19 0, 7, 14, 19 Unresponsive 0 0 Br eathing Slow 7, 19 7, 19 Coat Piloerection 0, 14 0, 14 Eyelids (Left) Partially Closed 0, 7, 19 0, 7, 19 Eyelids (Right) Partially Closed 0, 7, 19 0, 7, 19 278 TR Behaviour

Underactive 7, 14, 19 7, 14, 19 Unresponsive 7, 14 7, 14 Br eathing Slow 7, 19 7, 19 Coat Piloerection 14 14 Eyelids (Left) Partially Closed 7, 19 7, 19 Eyelids (Right) Partially Closed 7, 19 7, 19 W Killed for welfare reasons TR Total resorption192Huntingdon Life SciencesWAG0015

APPENDIX 3 - continu ed Request ID: 284712 Signs associated with dosing - individual observations for females after mating - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Days Group /Sex Animal Number Sign Pre exposure During dosing (~2 hours) Additional

observation During dosing

(~4 hours) 4F 279 W Behaviour

Underactive 0, 7, 14, 19 0, 7, 14, 19 Unresponsive 0, 14, 19 0, 14, 19 Br eathing Slow 0, 7, 14, 19 0, 7, 14, 19 Coat Piloerection 14, 19 14, 19 Eyelids (Left) Partially Closed 0, 7, 14, 19 0, 7, 14, 19 Eyelids (Right) P artially Closed 0, 7, 14, 19 0, 7, 14, 19 280 NP Behaviour

Underactive 7, 14, 19 7, 14, 19 Unresponsive 7, 14 7, 14 Br eathing Slow 7, 19 7, 19 Coat Piloerection 14 14 Eyelids (Le ft) Partially Closed 7, 19 7, 19 Eyelids (Right) Partially Closed 7, 19 7, 19 W Killed for welfare reasons NP Not pregnant193Huntingdon Life SciencesWAG0015

APPENDIX 3 - continued Request ID: 284713 Signs associated with dosing - individual ob servations for females during lactation - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Days Group /Sex Animal Number Sign During dosing (~2 hours) During dosing

(~4 hours) 3F 261 Behaviour

Underactive 9 9 264 Behaviour Underactive 9 9 268 Behaviour Underactive 5, 9 5, 9 Coat Piloerection 5 5 269 Behaviour Underactive 5, 9 5, 9 Coat Piloerection 5 5194Huntingdon Life SciencesWAG0015

APPENDIX 4 Request ID: 284710 Signs associated with dosing - individual observations prior to mating, durin g gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 2F 251 T Treatment Behaviour Un deractive 1 - 7 Unresponsive 1, 3 Coat Piloerection 2 - 7, 11, 14 Eyelids (Left) Partially Closed 14 Eyelids (Right) Partially Closed 14 Gestation Behaviour

Un deractive 7 Coat Piloerection 7, 19 Lactatio n No abnormalities detected T Terminal sacrifice195Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 2F 252 T Treatment Behaviour Un deractive 1 - 7 Unresponsive 1, 3 Coat Piloerection 2 - 7, 11, 14 Eyelids (Left) Partially Cl osed 14 Eyelids (Right) Partially Closed 14 Gestation Behaviour

Un deractive 0, 7, 14 Coat Piloerection 0, 7, 19 Lactatio n No abnormalities detected T Terminal sacrifice196Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 2F 253 T Treatment Behaviour Un deractive 1 - 7 Unresponsive 1, 3 Coat Piloerection 2 - 7, 11, 14 Eyelids (Left) Partially Closed 14 Eyelids (Right) Partially Closed 14 Gestation Coat

Piloerection 7 Lactatio n No abnormalities detected T Terminal sacrifice197Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 2F 254 T Treatment Behaviour Un deractive 1 - 7 Unresponsive 1, 3 Coat Piloerection 2 - 7, 11, 14 Eyelids (Left) Partially Closed 14 Eyelids (Right) Partially Closed 14 Gestation Coat

Piloerection 7 Lactatio n No abnormalities detected T Terminal sacrifice198Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 2F 255 T Treatment Behaviour Un deractive 1 - 7, 25 Unresponsive 1, 3 Coat Piloerecti on 2 - 7, 11, 14, 25 Eyelids (Left) Partially Closed 14 Eyelids (Right) Partially Closed 14 Gestation Behaviour

Un deractive 0, 14 Coat Piloerection 0, 14 Lactatio n No abnormalities detected T Terminal sacrifice199Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 2F 256 T Treatment Behaviour Un deractive 1 - 7 Unresponsive 1, 3 Coat Piloerection 2 - 7, 11, 14 Eyelids (Left) Partially Closed 14 Eyelids (Right) Partially Closed 14 Gestation Coat

Piloerection 7 Lactatio n No abnormalities detected T Terminal sacrifice200Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 2F 257 NP T Treatment Behaviour Un deractive 1 - 7 Unresponsive 1, 3 Coat Piloerection 2 - 7, 11, 14 Eyelids (Left) Partially Closed 14 Eyelids (Right) Partially Closed 14 Gestation Coat

Piloerection 7 T Terminal sacrifice NP Not pregnant201Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropen e 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 2F 258 T Treatment Behaviour Un deractive 1 - 7 Unresponsive 1, 3 Coat Piloerection 2 - 7, 11, 14 Eyelids (Left) Partially Closed 14 Eyelids (Right) Partially Closed 14 Gestation Behaviour

Un deractive 0, 7, 14 Coat Piloerection 0, 7, 19 Lactatio n No abnormalities detected T Terminal sacrifice202Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 2F 259 T Treatment Behaviour Un deractive 1 - 7 Unresponsive 1, 3 Coat Piloerection 2 - 7, 11, 14 Eyelids (Left) Partially Closed 14 Eyelids (Right) Partially Closed 14 Gestation Behaviour

Un deractive 7 Coat Piloerection 7, 19 Lactatio n No abnormalities detected T Terminal sacrifice203Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation f orfemales - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day( s) 2F 260 T Treatment Behaviour Un deractive 1 - 7 Unresponsive 1, 3 Coat Piloerection 2 - 7, 11, 14 Eyelids (Left) Partially Closed 14 Eyelids (Right) Partially Closed 14 Gestation Behaviour

Un deractive 7 Coat Piloerection 7, 19 Lactatio n No abnormalities detected T Terminal sacrifice204Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Ani mal Death /Sex Number Code Phase Sign Day (s) 3F 261 T Treatment Behaviour Un deractive 1 - 7, 11, 14, 18 Unresponsive 1 - 3, 6 - 7, 18 Br eathing Sh allow 14 Coat Piloerection 1 - 7, 14, 18 Eyelids (Left) Partially Closed 1, 3 - 5, 11, 14 Eyelids (Right) Partially Closed 1, 3 - 5, 11, 14 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1 Gestation Behaviour

Un deractive 7, 14, 19 Unresponsive 7 Br eathing Slow 7, 19 Coat Piloerection 14 Eyelids (Left) Partially Closed 19 T Terminal sacrifice205Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (pp m) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 261 T Gestation Eyelids (Right) Partially Closed 19 Lactatio n Behaviour

Un deractive 9 T Terminal sacrifice206Huntingdon Life SciencesWAG0015

APPENDIX 4 - continue d Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropen e 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 262 W Treatment Behaviour Un deractive 1 - 7, 11, 14, 18 Unresponsive 1 - 3, 6 - 7, 18 Br eathing Sh allow 14 Coat Piloerection 1 - 7, 14, 18 Eyelids (Left) Partially Closed 1, 3 - 5, 11, 14 Eyelids (Right) Partially Closed 1, 3 - 5, 11, 14 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1 Gestation Behaviour

Un deractive 0, 7, 14, 19 Unresponsive 7 Br eathing Slow 0, 14, 19 Coat Piloerection 7, 14, 19 Eyelids (Left) Partially Closed 0, 7, 14, 1 9 W Killed for welfare reasons207Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Contr ol 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 262 W Gestation Eyelids (Right) Partially Closed 0, 7, 14, 19 W Killed for welfare reasons208Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 263 U TR T Treatment Behaviour Un deractive 1 - 7, 11, 14, 18 Unresponsive 1 - 3, 6 - 7, 18 Br eathing Sh allow 14 Coat Piloerection 1 - 7, 14, 18 Eyelids (Left) Partially Closed 1, 3 - 5, 11, 14 Eyelids (Right) P artially Closed 1, 3 - 5, 11, 14 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1 Gestation Behaviour

Un deractive 0, 7, 14, 19 Unresponsive 0 Br eathing Slow 0, 7, 14, 19 Coat Piloerection 19 Eyelids (Left) Partially Closed 0, 7, 14 U Un ilateral implantation T Terminal sacrifice TR Total resorption209Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 263 U TR T Gestation Eyelids (Right) Partially Closed 0, 7, 14 U Un ilateral implantation T Terminal sacrifice TR Total resorption210Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID : 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3, 3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 264 T Treatment Behaviour Un deractive 1 - 7, 11, 14, 18 Unresponsive 1 - 3, 6 - 7, 18 Br eathing Sh allow 14 Coat Piloerection 1 - 7, 14, 18 Eyelids (Left) Partially Closed 1, 3 - 5, 11, 14 Eyelids (Right) Partially Closed 1, 3 - 5, 11, 14 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1 Gestation Behaviour

Un deractive 0, 7, 14, 19 Unresponsive 7 Br eathing Slow 0, 14, 19 Coat Piloerection 7, 14, 19 Eyelids (Left) Partially Closed 0, 7, 14, 19 T Terminal sacrifice211Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - tri fluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 264 T Gestation Eyelids (Right) Partially Closed 0, 7, 14, 19 Lactatio n Behaviour

Un deractive 9 T Terminal sacrifice212Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generati on Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 265 TLL Treatment Behaviour Un deractive 1 - 7, 11, 14 Unresponsive 1 - 3, 6 - 7 Br eathing Sh allow 14 Coat Piloerection 1 - 7, 14 Eyelids (Left) Partially Closed 1, 3 - 5, 11, 14 Eyelids (Right) Partially Closed 1, 3 - 5, 11, 14 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1 Gestation Behaviour

Un deractive 0, 7, 14, 19 Br eathing Sh allow 0 Slow 7, 19 Coat Piloerection 0, 14, 19 Eyelids (Left) Partially Closed 7, 19 TLL Total litter loss213Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase S ign Day (s) 3F 265 TLL Gestation Eyelids (Right) Partially Closed 7, 19 TLL Total litter loss214Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 266 TLL Treatment Behaviour Un deractive 1 - 7, 11, 14, 18, 21 Unresponsive 1 - 3, 6 - 7, 18, 21 Br eathing Sh allow 14 Coat Piloerection 1 - 7, 14, 18, 21 Eyelids (Left) Partially Closed 1, 3 - 5, 11, 14 Eyelids (Right) Partially Closed 1, 3 - 5, 11, 14 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1 Gestation Behaviour

Un deractive 0, 7, 14, 19 U nresponsive 19 Br eathing Slow 0, 14, 19 Coat Piloerection 7 Eyelids (Left) Partially Closed 0, 14, 19 TLL Total litter loss215Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (pp m) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 266 TLL Gestation Eyelids (Right) Partially Closed 0, 14, 19 TLL Total litter loss216Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 267 TLL Treatment Behaviour Un deractive 1 - 7, 11, 14, 18 Unresponsive 1 - 3, 6 - 7, 18 Br eathing Sh allow 14 Coat Piloerection 1 - 7, 14, 18 Eyelids (Left) Partially Closed 1, 3 - 5, 11, 14 Eyelids (Right) Partially Closed 1, 3 - 5, 11, 14 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1 Gestation Behaviour

Un deractive 7, 14, 19 Unresponsive 7 Br eathing Slow 7, 19 Coat Piloerection 14 Eyelids (Left) Partially Closed 19 TLL Total litter loss217Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - bromo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 267 TLL Gestation Eyelids (Right) Partially Closed 19 TLL Total litter loss218Huntingdon Life SciencesWAG0015

APPENDIX 4 - continu ed Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 268 T Treatment Behaviour Un deractive 1 - 7, 11, 14, 18 Unresponsive 1 - 3, 6 - 7, 18 Br eath ing Sh allow 14 Coat Piloerection 1 - 7, 14, 18 Eyelids (Left) Partially Closed 1, 3 - 5, 11, 14 Eyelids (Right) Partially Closed 1, 3 - 5, 11, 14 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1 Gestation Behaviour

Un deractive 7, 14, 19 Unresponsive 7 Br eathing Slow 7, 19 Coat Piloerection 14 Eyelids (Left) Partially Closed 19 T Terminal s acrifice219Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 268 T Gestation Eyelids (Right) Partially Closed 19 Lactatio n Behaviour

Un deractive 5, 9 Coat Piloerection 5 T Terminal sacrifice220Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for fem ales - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 269 T Treatment Behaviour Un deractive 1 - 7, 11, 14, 18 Unresponsive 1 - 3, 6 - 7, 18 Br eathing Sh allow 14 Coat Piloerection 1 - 7, 14, 18 Eyelids (Left) Partially Closed 1, 3 - 5, 11, 14 Eyelids (Right) Partially Closed 1, 3 - 5, 11, 14 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1 Gestation Behaviour

Un deractive 0, 7, 14, 19 Unresponsive 0 Br eathing Slow 0, 7, 14, 19 Coat Piloerection 19 Eyelids (Left) Partially Closed 0, 7, 14 T Terminal sacrifice221Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 269 T Gestation Eyelids (Right) Partially Closed 0, 7, 14 Lactatio n Behaviour

Un deractive 5, 9 Coat Piloerection 5 T Terminal sacrifice222Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trif luoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 270 TLL Treatment Behaviour Un deractive 1 - 7, 11, 14, 18 Unresponsive 1 - 3, 6 - 7, 18 Br eathing Sh allow 14 Coat Piloerection 1 - 7, 14, 18 Eyelids (Left) Partially Closed 1, 3 - 5, 11, 14 Eyelids (Right) Partially Closed 1, 3 - 5, 11, 14 Eye (Left) Lachrymation 1 Eye (Right) Lachrymation 1 Gestation Behaviour

Un deractive 7, 14, 19 Unresponsive 7 Br eathing Slow 7, 19 Coat Piloerection 14 Eyelids (Left) Partially Closed 19 TLL Total litter loss223Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - t rifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 3F 270 TLL Gestation Eyelids (Right) Partially Closed 19 TLL Total litter loss224Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3 ,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 4F 271 TR T Treatment Behaviour Un deractive 1 - 8, 11, 1 4, 18, 21 Unresponsive 1 - 2, 4 - 8, 11, 14, 18, 21 Br eathing Sh allow 1 - 7, 14 Coat Piloerection 1 - 8, 14, 18, 21 Eyelids (Left) Partially Closed 1, 3 - 8, 11, 14, 18, 21 Eyelids (Right) Partially Closed 1, 3 - 8, 11, 14, 18, 21 Posture Hunched 8 Staining Other Colour (see comment) 8 Comments: brown staining head 8 Gestation Behaviour

Un deractive 0, 7, 14, 19 Unresponsive 0, 19 Br eathing Slow 7, 14, 19 Coat Piloerection 0, 7, 14, 19 T Terminal sacrifice TR Total resorption225Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal De ath /Sex Number Code Phase Sign Day (s) 4F 271 TR T Gestation Eyelids (Left) Partially Closed 0, 7, 14, 19 Eyelids (Right) Partially Closed 0, 7, 14, 19 Muscle Reaction Reduced Body Tone 14 272 FTM T Treatment Behaviour Un deractive 1 - 7, 11, 14, 18, 21, 25, 28, 32, 35, 40, 47 Unresponsive 1 - 2, 4 - 7, 11, 14, 18, 21, 25, 32, 40, 47 Br eathing Sh allow 1 - 7, 14 Slow 25, 28, 35, 40 Coat Piloerection 1 - 7, 14, 18, 21, 28, 32, 40, 47 Eyelids (Left) Partially Closed 1, 3 - 7, 11, 14, 18, 21, 25, 28, 35, 40, 47 Eyelids (Right) Partially Closed 1, 3 - 7, 11, 14, 18, 21, 25, 28, 35, 40, 47 T Terminal sacrifice TR Total resorption FTM Failed to mate226Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 4F 273 TLL Treatment Behaviour Un deractive 1 - 8 , 11, 14 Unresponsive 1 - 2, 4 - 8, 11, 14 Br eathing Sh allow 1 - 7, 14 Coat Piloerection 1 - 8, 14 Eyelids (Left) Partially Closed 1, 3 - 8, 11, 14 Eyelids (Right) Partially Closed 1, 3 - 8, 11, 14 Posture Hunched 8 Staining Other Colour (see comment) 8 Comments: brown staining head 8 Gestation Behaviour

Un deractive 0, 7, 14, 19 Unresponsive 0, 14 Br eathing S low 0, 7, 19 Coat Piloerection 0, 7, 14 TLL Total litter loss227Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 4F 273 TLL Gestation Eyelids (Left) Partially Closed 0, 7, 14, 19 Eyelids (Right) Partially Closed 0, 7, 14, 19 TLL Total litter loss228Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 290 0 Group Animal Death /Sex Number Code Phase Sign Day (s) 4F 274 TR T Treatment Behaviour Un deractive 1 - 7, 11, 14 Unresponsive 1 - 2, 4 - 7, 11, 14 Br eathing Sh allow 1 - 7, 14 Coat Piloerection 1 - 7 , 14 Eyelids (Left) Partially Closed 1, 3 - 7, 11, 14 Eyelids (Right) Partially Closed 1, 3 - 7, 11, 14 Gestation Behaviour

Un deractive 0, 7, 14, 19 Unresponsive 0, 14 Coat Piloerection 0 , 7, 19 Eyelids (Left) Partially Closed 0, 7 Eyelids (Right) Partially Closed 0, 7 Posture Hunched 14 T Terminal sacrifice TR Total resorption229Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs asso ciated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 4F 275 FTM T Treatment Behaviour Un deractive 1 - 8, 11, 14, 18, 21, 25, 28, 32, 35, 40, 47 Unresponsive 1 - 2, 4 - 8, 11, 14, 18, 21, 25, 32, 40, 47 Br eathing Sh allow 1 - 7, 14 Slow 25, 28, 35, 40 Coat Piloerection 1 - 8, 14, 18, 21, 28, 32, 40, 47 Eyelids (Left) Partially Closed 1, 3 - 8, 11, 14, 18, 21, 25, 28, 35, 40, 47 Eyelids (Right) Partially Closed 1, 3 - 8, 11, 14, 18, 21, 25, 28, 35, 40, 47 Posture Hunched 8 Staining Other Colour (see comment) 8 Comments: Brow n staining head 8 T Terminal sacrifice FTM Failed to mate230Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 4F 276 W Treatment Behaviour Un deractive 1 - 7, 11, 14, 18, 21 Unresponsive 1 - 2, 4 - 7, 11, 14, 18, 21 Br eathing Sh allow 1 - 7, 14 Coat Piloerection 1 - 7, 14, 18, 21 Eyelids (Left) Partially Closed 1, 3 - 7, 11, 14, 18, 21 Eyelids (Right) Partially Closed 1, 3 - 7, 11, 14, 18, 21 Gestation Behaviour

Un deractive 0, 7, 14, 19 Unresponsive 0 Br eathing Slow 7, 19 Coat Piloerection 0, 14 Eyelids (Left) Partially Closed 0, 7, 19 Eyelids (Right) Partially Closed 0, 7, 19 W Killed for welfare reasons231Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Grou p : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 4F 277 FTM T Treatment Behaviour Un deractive 1 - 7, 11, 14, 18, 21, 25, 28, 32, 35, 40, 47 Unresponsive 1 - 2, 4 - 7, 11, 14, 18, 21, 25, 32, 40, 47 Br eathing Sh allow 1 - 7, 14 Slow 25, 28, 35, 40 Coat Piloerection 1 - 7, 1 4, 18, 21, 28, 32, 40, 47 Eyelids (Left) Partially Closed 1, 3 - 7, 11, 14, 18, 21, 25, 28, 35, 40, 47 Eyelids (Right) Partially Closed 1, 3 - 7, 11, 14, 18, 21, 25, 28, 35, 40, 47 T Terminal sacrifice FTM Failed to mat e232Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 4F 278 TR T Treatment Behaviour Un deractive 1 - 7, 11, 14, 18 Unresponsi ve 1 - 2, 4 - 7, 11, 14, 18 Br eathing Sh allow 1 - 7, 14 Coat Piloerection 1 - 7, 14, 18 Eyelids (Left) Partially Closed 1, 3 - 7, 11, 14, 18 Eyelids (Right) Partially Closed 1, 3 - 7, 11, 14, 18 Gestation Behaviour

Un deractive 7, 14, 19 Unresponsive 7, 14 Br eathing Slow 7, 19 Coat Piloerection 14 Eyelids (Left) Partially Closed 7, 19 Eyelids (Right) Partially Closed 7, 19 T Terminal sacrifice TR Total resorption233Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 4F 279 W Treatment Behaviour Un deractive 1 - 7, 11, 14, 18, 21, 25 Unresponsive 1 - 2, 4 - 7, 11, 14, 18, 21, 25 Br eathing Sh allow 1 - 7, 14 Slow 25 Coat Piloerection 1 - 7, 14, 18, 21 Eyelids (Left) Partially Closed 1, 3 - 7, 1 1, 14, 18, 21, 25 Eyelids (Right) Partially Closed 1, 3 - 7, 11, 14, 18, 21, 25 Gestation Behaviour

Un deractive 0, 7, 14, 19 Unresponsive 0, 14, 19 Br eathing Slow 0, 7, 14, 19 Coat Pilo erection 14, 19 Eyelids (Left) Partially Closed 0, 7, 14, 19 Eyelids (Right) Partially Closed 0, 7, 14, 19 W Killed for welfare reasons234Huntingdon Life SciencesWAG0015

APPENDIX 4 - continued Request ID: 284710 Signs associated with dosing - individual observations prior to mating, during gestation and lactation for females - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Death /Sex Number Code Phase Sign Day (s) 4F 280 NP T Treatment Behaviour Un deractive 1 - 7, 11, 14, 18 Unresponsive 1 - 2, 4 - 7, 11, 14, 18 Br eathing Sh allow 1 - 7, 14 Coat Piloerection 1 - 7, 14, 18 Eyelids (Left) Partially Closed 1, 3 - 7, 11, 14, 18 Eyelids (Right) Partially Closed 1, 3 - 7, 11, 14, 18 Gestation Behaviour

Un deractive 7, 14, 19 Unresponsive 7, 14 Br eathing Slow 7, 19 Coat Piloerection 14 Eyelids (Left) Partially Closed 7, 19 Eyelids (Right) Partially Closed 7, 19 T Terminal sacrifice NP Not pregnant235Huntingdon Life SciencesWAG0015

APPENDIX 5 Request ID: 284396 Bodyweight - individual values for males - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal D ay Day Day Day Day Day Day Day Day Day Day Day /Sex Number - 7 1 8 11 15 22 29 33 36 39 43 46 1 M 201 257 327 377 394 410 414 460 477 490 495 510 522

202 248 318 363 380 405 411 438 442 456 453 463 476 203 268 332 373 372 409 431 446 458 457 468 487 499 204 255 328 375 389 416 447 478 499 514 516 533 541 205 270 349 401 424 450 478 506 521 532 530 538 553 206 249 332 402 422 462 487 515 539 545 551 563 574 207 257 322 373 386 404 433 453 471 484 495 502 512 208 259 328 371 398 414 450 477 488 504 512 521 536 209 259 339 391 409 433 467 493 507 520 528 547 555 21 0 269 319 374 381 401 421 450 472 475 479 501 510 2 M 211 256 327 384 400 424 457 477 485 499 503 490 496

212 251 304 326 335 345 372 382 388 392 391 399 401 213 253 320 360 369 396 422 448 466 475 483 490 504 214 274 338 408 432 458 495 522 533 547 547 551 561 215 239 303 362 388 410 434 457 474 487 487 500 510 216 262 339 389 412 438 468 505 510 518 526 526 533 217 25 8 321 365 377 388 406 425 432 436 441 453 461 218 247 313 338 347 362 379 402 413 415 426 427 432 219 248 308 325 332 341 360 397 400 411 415 423 431 220 255 322 351 354 370 405 411 417 423 429 429 434236Huntingdon Life SciencesWAG0015

APPENDIX 5 - continued Request ID: 284396 Bodyweight - individual values for males - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day /Sex Number 50 1 M 201 534

202 479 203 512 204 563 205 570 206 591 207 529 208 545 209 570 210 524 2 M 211 503

212 413 213 505 214 567 215 516 216 545 217 468 218 438 219 436 220 438237Huntingdon Life SciencesWAG0015

APPENDIX 5 - continued Request ID: 284396 Bodyweight - individual values for males - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day Day Day Day Day Day Day Day Day Day Day /Sex Number - 7 1 8 11 15 22 29 33 36 39 43 46 3 M 221 255 323 340 345 368 385 413 421 430 437 445 459

222 251 307 332 344 354 374 380 389 397 396 399 405 223 254 318 343 355 371 393 405 416 419 424 418 425 224 249 319 358 370 384 416 421 425 434 431 426 434 225 248 322 356 366 398 421 435 446 456 459 463 472 226 258 317 338 338 359 378 407 416 419 426 433 438 227 257 330 375 385 407 431 455 475 479 483 497 504 228 268 332 349 353 372 391 406 426 430 436 442 442 229 254 322 344 347 367 383 407 417 428 432 436 450 230 266 333 354 367 385 406 422 427 443 444 456 462 4 M 231 242 314 341 355 375 405 425 444 452 451 457 467

232 254 337 350 358 372 387 408 401 411 406 415 407 233 253 316 315 326 331 352 362 369 380 370 385 375 234 246 312 321 339 344 369 383 387 392 395 401 409 235 246 333 355 366 387 406 425 430 432 437 444 450 236 276 343 356 373 389 382 429 431 426 437 450 447 237 252 310 337 343 361 384 393 393 397 397 397 396 238 259 325 333 351 356 376 403 396 399 397 389 387 239 252 339 367 385 396 416 425 431 435 442 440 449 240 252 322 341 353 368 385 406 408 416 418 416 418238Huntingdon Life SciencesWAG0015

APPENDIX 5 - continued Request ID : 284396 Bodyweight - individual values for males - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day /Sex Number 50 3 M 221 462

222 412 223 431 224 443 225 478 226 444 227 510 228 451 229 449 230 475 4 M 231 468

232 406 233 380 234 410 235 457 236 459 237 403 238 385 239 447 240 423239Huntingdon Life SciencesWAG0015

APPENDIX 6 Request ID: 284397 Bodyweight - individual values for females before pairing - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - brom o - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day Day Day Day Day /Sex Number - 7 1 8 11 15 22 1 F 241 206 237 247 250 257

242 193 213 231 233 240 243 196 224 232 233 240 244 194 219 239 238 248 245 209 224 227 243 248 246 207 237 255 254 257 247 189 215 219 219 223 248 219 247 255 254 258 249 196 214 230 235 242 250 210 224 235 234 242 2 F 251 215 234 241 247 258

252 206 224 234 238 242 253 203 231 231 233 242 254 198 231 249 252 267 255 201 218 247 255 263 276 256 197 228 239 239 242 257 215 230 240 254 256 258 205 224 236 234 241 259 201 220 223 237 243 260 209 235 253 256 264240Huntingdon Life SciencesWAG0015

APPENDIX 6 - continued Request ID: 284397 Bodyweight - individual values for f emales before pairing - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day Day Day Day Day Day Day Day Day Day Day /Sex Number - 7 1 8 11 15 22 29 33 36 39 43 46 3 F 261 205 225 231 227 235

262 207 237 246 255 263 263 209 234 238 245 250 264 207 214 229 242 241 265 198 229 236 241 242 266 202 213 222 225 229 230 267 211 237 238 236 255 268 199 228 230 232 245 2 69 195 215 220 225 232 270 192 217 231 232 247 4 F 271 218 243 206 219 230

272FTM 214 239 238 237 230 236 245 243 255 247 254 254 273 208 235 217 224 219 274 197 204 230 238 235 275 FTM 205 230 238 241 237 236 235 248 257 247 249 251 276 202 223 222 232 237 216 277 FTM 212 226 238 233 224 241 241 245 251 24 2 244 248 278 202 221 226 233 241 279 195 226 221 226 229 242 280 218 233 235 231 234 FTM Failed to mate241Huntingdon Life SciencesWAG0015

APPENDIX 6 - continued Request ID: 284397 Bo dyweight - individual values for females before pairing - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Grou p Animal Day Day /Sex Number 50 53 4 F 272FTM 252 255

275 FTM 257 262 277 FTM 242 243 FTM Failed to mate242Huntingdon Life SciencesWAG0015

APPENDIX 7 Request ID: 284398 Bodyweight - individual values for females after mating - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day Day Day Day Day Day /Sex Number 0 3 7 10 14 17 20 1 F 241 262 276 298 302 320 349 399

242 246 254 268 284 304 328 373 243 245 259 276 292 310 340 390 244 253 277 293 299 327 375 423 245 247 264 300 303 322 352 392 246 273 284 316 320 351 389 451 247 236 253 266 285 300 337 384 248 271 286 301 313 337 364 416 249 257 282 304 322 347 370 438 250 267 293 317 323 356 392 434 2 F 251 260 272 286 292 311 338 390

252 243 261 269 283 297 334 380 253 248 268 283 295 307 329 369 254 280 289 305 324 351 388 451 255 266 297 320 334 354 390 446 256 249 267 272 259 294 339 394 257NP 269 273 288 285 288 293 293 258 244 257 267 280 299 308 338 259 243 260 280 292 306 328 371 260 264 275 293 308 331 371 433 NP Not pregnant, excluded from calculation of group means and statistical evaluation243Huntingdon Life SciencesWAG0015

APPENDIX 7 - continued Request ID: 284398 Bodyweight - individual values for females after mating - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day Day Day Day Day Day /Sex Number 0 3 7 10 14 17 20 3 F 261 249 255 264 281 303 332 379

262 267 284 296 295 332 351 395 263 U TR 266 284 286 297 308 282 287 264 246 268 286 297 326 347 401 265 237 259 274 285 303 322 355 266 TLL 234 263 276 280 302 305 331 267 252 271 276 291 309 327 372 268 254 263 281 286 313 339 389 269 249 257 266 279 298 324 372 270 256 266 278 291 320 350 399 4 F 271 TR 244 251 271 281 265 275 282

273 227 234 239 248 272 281 316 274 TR 238 258 264 281 293 307 320 276W 215 259 246 266 282 301 322 278 TR 256 258 272 275 302 304 300 279W 251 262 270 280 294 289 289 280NP 235 261 260 264 265 257 264 U Un ilateral implantation TR Total resorption, excluded from calculation of group means and statistical evaluation NP Not pregnant, excluded from calculation of group means and statistical evaluation W Killed for welfare reasons, excluded from calculation of group means and statistical evaluation TLL Total litter loss post partum , g estation data has been inc luded from calculation of group means and statistical evaluation244Huntingdon Life SciencesWAG0015

APPENDIX 8 Request ID: 284399 Bodyweight - individual values for females during lactation - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day Day /Sex Number 1 5 10 1 F 241 296 304 344

242 292 295 304 243 281 28 8 328 244 312 352 370 245 304 317 329 246 327 359 374 247 269 293 313 248 313 342 356 249 320 337 340 250 327 332 357 2 F 251 302 321 328

252 279 290 284 253 281 285 306 25 4 340 356 377 255 319 321 344 256 296 321 327 258 285 282 302 259 290 306 309 260 309 317 322245Huntingdon Life SciencesWAG0015

APPENDIX 8 - continued Request ID: 284399 Bodyweight - individual values for females during lactation - F0 generation (g) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day Day /Sex Number 1 5 10 3 F 261 284 298 312

262W 310 264 310 309 305 265 TLL 288 267 TLL 304 268 290 294 293 269 284 296 277 270 TLL 297 4 F 273 TLL 260 W Killed for welfare reasons, exc luded from calculation of group means and statistical evaluation TLL Total litter loss, excluded from calculation of group means and statistical evaluation246Huntingdon Life SciencesWAG0015

APPENDIX 9 Request ID: 284400 Food consumption - individual values for males and before pairing for females - F0 generation (g/animal/week) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Cage Week Week Week /Sex Number - 1 1 2 1M 41 201 213 176 42 213 228 188 2M 43 200 198 175 44 196 185 151 3M 45 198 168 143 46 203 156 140 4M 47 198 145 148 48 210 152 159 Week 2 consumption is over a 6 day period (g/animal/6 days).247Huntingdon Life SciencesWAG0015

APPENDIX 9 - continued Request ID: 284400 Food consumption - individual values for males and before pairing for females - F0 generation (g/animal/week) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Cage Week Week Week /Sex Number - 1 1 2 1F 49 135 132 129 50 139 134 132 2F 51 137 117 122 52 137 118 126 3F 53 137 101 115 54 136 102 113 4F 55 144 82 96 56 131 90 101248Huntingdon Life SciencesWAG0015

APPENDIX 10 Request ID: 284401 Food consumption - individual values for females after mating - F0 generation (g/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3 ,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day Day Day Day Day /Sex Number 0 - 2 3 - 6 7 - 9 10 - 13 14 - 16 17 - 19 1F 241 23 27 26 25 29 25 242 19 21 23 26 23 24 243 22 23 25 24 24 27 244 25 25 26 29 30 32 245 20 27 25 27 29 27 246 20 26 25 28 29 35 247 28 24 26 25 29 29 248 23 22 22 24 25 28 249 28 31 32 31 27 33 250 28 31 30 33 33 31 2F 251 19 20 19 22 22 28 252 20 19 21 22 22 27 253 21 22 22 23 23 28 254 23 24 25 28 30 34 255 27 25 22 26 29 30 256 24 16 14 27 29 36 257 NP 23 24 21 22 19 24 258 21 20 20 24 20 27 259 21 23 22 25 25 30 260 20 24 24 26 26 32 NP Not pregnant, excluded from calculation of group means and statistical evalua tion249Huntingdon Life SciencesWAG0015

APPENDIX 10 - continued Request ID: 284401 Food consumption - individual values for females after mating - F0 generation (g/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day Day Day Day Day /Sex Number 0 - 2 3 - 6 7 - 9 10 - 13 14 - 16 17 - 19 3F 261 18 20 22 21 23 29 262 23 21 22 27 26 30 263 U TR 19 17 20 21 16 21 264 23 25 23 25 26 31 265 21 24 24 22 22 28 266 TLL 21 21 19 24 22 28 267 22 20 20 21 20 29 268 20 22 23 25 25 30 269 20 20 21 24 24 27 270 23 24 28 26 26 28 4F 271 TR 18 20 20 19 15 20 273 15 16 17 20 21 29 274 TR 21 21 21 23 24 24 276 W 20 16 1 8 19 24 26 278 TR 20 22 21 23 22 23 279 W 20 21 21 23 23 22 280 NP 23 21 20 17 18 18 U TR W NP TLL Un ilateral implantation Total resorption, excluded from calculation of group means and statistical evaluation Killed for welfare reasons, ex cluded from calculation of group means and statistical evaluation Not pregnant, excluded from calculation of group means and statistical evaluation Total litter loss post partum , gestation data has been inc luded from calculation of group means and statistical evaluation250Huntingdon Life SciencesWAG0015

APPENDIX 11 Request ID: 284402 Food consumption - individual values for females during lactation - F0 generation (g/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day /Sex Number 1 - 4 5 - 9 1F 241 37 54 242 33 47 243 40 53 244 48 64 245 39 51 246 44 64 247 39 55 248 38 58 249 43 57 250 35 57 2F 251 39 4 4 252 27 31 253 31 42 254 36 45 255 28 41 256 39 42 258 22 27 259 29 35 260 32 40251Huntingdon Life SciencesWAG0015

APPENDIX 11 - continued Request ID: 284402 Food consumption - individual values for females during lactation - F0 generation (g/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day /Sex Number 1 - 4 5 - 9 3F 261 32 33 264 21 23 268 29 32 269 31 33252Huntingdon Life SciencesWAG0015

APPENDIX 12 Request ID: 287346 Water consumption - individual values for males before and after pairing and females before mating - F0 generation (mL/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Cage Day Day Day Day Day Day Day Day Day Day Day Day Day /Sex Number 9 10 11 32 33 34 35 39 43 44 45 46 47 1M 41 40 4 3 42 41 42 36 39 39 43 35 41 41 39 42 43 50 47 44 47 40 37 45 44 42 44 42 41 2M 43 48 52 47 44 43 40 41 45 39 41 45 42 41 44 35 41 35 37 34 33 30 34 31 34 36 33 32 3M 45 43 44 40 43 38 38 38 42 38 39 44 36 42 46 44 4 8 46 45 41 39 36 42 38 39 44 42 39 4M 47 45 50 48 42 40 39 37 42 39 40 43 42 42 48 45 49 48 41 38 34 35 35 34 36 43 36 34253Huntingdon Life SciencesWAG0015

APPENDIX 12 - continued Request ID: 287346 Water consumption - individual values for ma les before and after pairing and females before mating - F0 generation (mL/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Cage Day /Sex Number 48 1M 41 39 42 41 2M 43 38 44 31 3M 45 36 46 38 4M 47 42 48 35254Huntingdon Life SciencesWAG0015

APPENDIX 12 - continued Request ID: 287346 Water consumption - individual values for males before and after pairin g and females before mating - F0 generation (mL/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Cage Day Day D ay Day Day Day Day Day Day Day Day Day /Sex Number 9 10 11 33 34 35 39 43 44 45 46 47 1F 49 22 37 29 50 23 32 32 2F 51 30 41 30 52 29 43 31 3F 53 36 56 38 5 4 40 61 41 4F 55 54 64 52 34 36 26 33 30 32 48 34 37 56 36 54 39 7 8 14 14 11 12 10 10 8 Group 4F, data presented post Day 11 for females that didn’t mate255Huntingdon Life SciencesWAG0015

APPENDIX 12 - continued Request ID: 287346 Water consumption - individual values for males before and after pairing and females before mating - F0 generation (mL/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Cage Day Day /Sex Number 48 53 1F 49 50 2F 51 52 3F 53 54 4F 55 30 41 56 10 13 Group 4F, data presented post Day 11 for females that didn’t mate256Huntingdon Life SciencesWAG0015

APPENDIX 13 Request ID: 284865 Water consumption - individual values for females after mating - F0 generation (mL/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluorop ropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day Day Day Day Day Day Day Day Day Day Day Day /Sex Number 0 1 2 3 4 5 6 7 8 9 10 11 12 1F 241 56 71 66 49 54 62 62 61 61 59 72 62 60 242 59 82 71 52 63 68 61 62 55 65 78 65 66 243 35 42 52 80 55 46 61 60 62 58 59 67 @ 244 60 67 66 36 33 43 38 41 47 38 84 65 67 245 49 61 68 38 60 52 66 58 64 54 52 39 48 246 6 63 52 63 48 65 61 53 73 68 77 75 76 247 70 70 40 40 43 42 43 48 41 58 55 55 48 248 57 68 66 59 55 66 59 63 56 75 68 68 74 249 63 77 76 48 53 73 52 74 58 62 63 65 66 250 56 67 67 54 62 61 57 64 58 71 71 70 74 2F 251 70 79 77 54 67 69 71 68 56 69 62 55 62 252 42 49 80 79 76 48 76 72 69 71 74 79 87 253 56 85 107 94 57 70 72 77 74 91 86 54 50 254 31 51 81 65 42 68 54 62 60 49 61 84 77 255 49 55 55 96 85 91 88 91 87 65 79 86 98 256 41 43 44 36 45 66 41 72 88 82 89 85 78 257 NP 32 36 47 44 31 57 43 53 34 37 40 69 55 258 29 36 54 70 58 36 54 54 55 58 62 59 65 259 43 62 59 54 54 54 60 62 58 62 75 77 75 260 30 40 35 51 65 69 58 65 56 64 76 70 71 NP @ Not pregnant, excluded from calculation of group means and statistical evaluation Water bottle leaked257Huntingdon Life SciencesWAG0015

APPENDIX 13 - continued Request ID: 284865 Water consumptio

n - individual values for females after mating - F0 generation (mL/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day Day Day Day Day /Sex Number 13 14 15 16 17 18 1F 241 71 71 76 71 72 70 242 69 74 71 66 77 69 243 75 60 71 71 69 67 244 70 73 83 76 87 86 245 48 52 48 49 71 75 246 67 57 78 82 63 63 247 70 65 57 65 62 83 248 7 5 67 62 79 82 79 249 64 58 67 62 80 83 250 80 70 68 60 57 60 2F 251 66 63 65 67 82 76 252 77 81 85 68 88 72 253 59 63 63 85 82 78 254 78 85 55 59 52 76 255 85 87 91 112 82 57 256 89 86 95 96 87 68 257 NP 53 46 58 53 49 40 258 67 64 73 80 78 72 259 90 88 86 89 71 57 260 84 86 77 83 57 50 NP Not pregnant, excluded from calculation of group means and statistical evaluation258Huntingdon Life SciencesWAG0015

APPENDIX 13 - continued Request ID: 284865 Water consumption - individual values for femal es after mating - F0 generation (mL/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day Day Day Day Day Day Day Day Day Day Day Day /Sex Number 0 1 2 3 4 5 6 7 8 9 10 11 12 3F 261 50 52 63 53 65 68 71 75 79 77 89 85 75 262 46 41 38 39 27 54 40 44 41 77 75 76 83 263 U TR 37 56 67 62 69 68 71 72 94 79 66 80 81 264 68 70 74 64 58 73 64 73 70 55 77 86 36 265 44 59 87 82 64 56 83 77 80 96 86 79 90 266 63 77 78 91 82 105 70 61 67 74 78 73 69 267 77 89 85 72 57 64 60 61 63 58 85 93 83 268 43 56 58 56 43 51 47 49 55 54 91 83 89 269 35 49 58 58 66 60 55 63 62 64 66 66 66 270 60 75 63 40 66 56 68 62 59 66 72 65 66 4F 271 TR 74 76 70 81 75 84 72 85 70 79 63 80 68 273 34 42 75 80 68 51 70 73 68 74 57 79 43 274 TR 56 83 96 85 65 60 68 65 76 64 64 106 @ 276 W 72 58 63 60 53 64 72 75 67 80 82 86 67 278 TR 60 82 65 53 55 49 44 44 55 37 89 71 80 279 W 64 39 74 63 55 68 65 57 42 75 80 83 87 280 NP 74 82 80 60 66 69 73 79 78 69 93 70 79 U Un ilateral implantation TR Total resorption, excluded from calculation of group means and statistical evaluation W Killed for welfare reasons, excluded from calculation of group means and statistical evaluation NP Not pregnant, excluded from calculation of group means and statistical evaluation @ Water bottle leaked259Huntingdon Life SciencesWAG0015

APPENDIX 13 - continued Request ID : 284865 Water consumption - individual values for females after mating - F0 generation (mL/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day Day Day Day Day /Sex Number 13 14 15 16 17 18 3F 261 96 84 93 87 86 86 262 90 71 69 90 79 85 263 U TR 74 59 76 47 79 80 264 68 51 54 84 80 82 265 93 89 94 96 104 86 266 102 98 108 97 102 1 07 267 98 100 113 100 103 105 268 94 96 96 91 100 92 269 61 65 62 86 80 72 270 79 84 87 86 82 84 4F 271 TR 55 66 83 82 82 100 273 44 47 74 53 49 47 274 TR 89 62 116 107 98 87 276 W 95 82 89 94 87 89 278 TR 80 91 89 81 56 47 279 W 78 83 91 91 49 74 280 NP 89 95 72 80 47 42 U Un ilateral implantation TR Total resorption, excluded from calculation of group means and statistical evaluation W Killed for welfare reasons, excluded from calculation of group means and stati stical evaluation NP Not pregnant, excluded from calculation of group means and statistical evaluation260Huntingdon Life SciencesWAG0015

APPENDIX 14 Request ID: 284866 Water consumption - individual values for females during lactation - F0 generation (mL/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day Day Day Day Day Day Day Day /Sex Number 1 2 3 4 5 6 7 8 9 1F 241 53 95 56 66 67 100 84 106 110 242 46 103 45 62 80 86 75 94 66 243 43 51 66 121 92 78 88 108 79 244 52 142 74 94 102 108 80 107 83 245 55 82 42 61 86 74 81 94 106 246 44 62 73 107 82 107 112 125 117 247 104 47 67 61 69 79 78 93 108 248 101 56 66 83 95 87 99 100 109 249 103 57 76 75 107 85 106 114 105 250 111 51 60 65 102 81 101 107 120 2F 251 145 44 67 58 72 57 80 84 106 252 39 46 112 48 46 46 69 59 88 253 43 96 40 53 74 95 87 120 114 254 43 104 52 66 80 67 75 83 95 255 18 68 44 43 80 56 85 98 64 256 58 116 61 75 58 256 77 95 82 258 34 135 32 31 59 51 46 53 69 259 95 37 53 46 61 52 83 76 103 260 89 41 55 51 74 59 81 86 114261Huntingdon Life SciencesWAG0015

APPENDIX 14 - continued Request ID: 284866 Water consumption - individual values for females during lactation - F0 generation (mL/animal/day) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day Day Day Day Day Day Day Day Day /Sex Number 1 2 3 4 5 6 7 8 9 3F 261 39 51 56 79 53 76 83 95 93 264 26 32 35 64 34 52 61 75 53 268 38 43 57 49 65 106 83 89 88 269 30 49 66 62 60 45 88 86 62 270 TLL 96 4F 273 TLL 111 TLL Total litter loss, excluded from calculation of group means and statistical evaluation262Huntingdon Life SciencesWAG0015

APPENDIX 15 Oestrous cycle length , mating performance, fertility and gestation length - individual values - F0 generatio n Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Number of Sperm Individual Pre - coital Gestation Female copulation count cycle length interval Male length Group number plugs category (days) (days) number Pregnancy (days) 1 241 4 2 4,4,4,4 3 201 + 22

242 4 4 4,4,4,4 3 202 + 22 243 4 4 4,4,4 1 203 + 22.5 244 6 4 4,4,4,4 3 204 + 22.5 245 4 4 5^,5,5,4 3 205 + 22.5 246 3 4 5^,5,5,5 5 206 + 22 247 5 0 4,4,4,4 4 207 + 22 248 3 4 4,4,4,4 4 208 + 22 249 4 3 4,4,4,4 3 209 + 23 250 4 3 4,4,4,4 4 210 + 22

 0 No sperm 1 Occasional sperm 2 Continuous few sperm 3 Many scattered sperm 4 Solid masses of sperm ^ Estimated minimum cycle length - cycle in progress at start or end of smearing period263Huntingdon Life SciencesWAG0015

APPENDIX 15 - continued Oestrous cycle length, mating performance, fertility and gestation length - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Number of Sperm Individual Pre - coital Gestation Female copulation count cyc le length interval Male length Group number plugs category (days) (days) number Pregnancy (days) 2 251 4 3 5,5,5 3 211 + 23.5

252 4 3 4,5,5 1 212 + 23 253 4 2 4,5,5 2 213 + 23.5 254 4 1 5,5,5 2 214 + 23 255 1 2 5,4,5,5,5 10 215 + 23.5 256 5 3 5,5,5 2 216 + 23 257 3 1 5,5,4 2 217 - - 258 2 3 4,4,5 1 218 + 23.5 259 4 4 5,5,5 3 219 + 23.5 260 4 1 4,4,4,4 3 220 + 23

 0 No sperm 1 Occasional sperm 2 Continuous few sperm 3 Many scattered sperm 4 Solid masses of sperm264Huntingdon Life SciencesWAG0015

APPENDIX 15 - continued Oestrous cycle length, mating performance, fertility and gestation length - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Number of Sperm Individual Pre - coital Gestation Female copulation count cycle length interval Male length Group number plugs category (days) (days) number Pregnancy (days) 3 261 3 0 5,5,5 3 221 + 23.5

262 3 3 5^,5,5,5 4 222 + 24.5 263NLL 4 4 7,6,5 5 223 + - 264 3 4 12,5 4 224 + 23.5 265 2 3 6,4,4 1 225 + 25.5 266 3 0 5,5,6,6 8 226 + 25 267 3 1 5,6,5 3 227 + 25.5 268 5 0 5,5,5 3 228 + 24.5 269 3 0 5^,5,5,5 5 2 29 + 23.5 270 1 2 4,4,4,4 3 230 + 23

 0 No sperm 1 Occasional sperm 2 Continuous few sperm 3 Many scattered sperm 4 Solid masses of sperm ^ Estimated minimum cycle length - cycle in progress at start or end of smearing period NLL No live litter

– pr egnancy indicated by implantatio n sites265Huntingdon Life SciencesWAG0015

APPENDIX 15 - continued Oestrous cycle length, mating performance, fertility and gestation length - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Number of Sperm Individual Pre - coital Gestation Female copulation count cycle length interval Male length Group number plugs category (days) (days) number Pregnancy (days) 4 271NLL 2 3 12,6 6 231 + -

272 - - 15^,6,9^ FTM - - - 273 4 3 14 1 233 + 25.5 274NLL 4 3 16 2 234 + - 275 - - 7,7,5,2,8^ FTM - - - 276NLL* 2 2 6,4,9e 7 236 + - 277 - - 16,5,6 FTM - - - 278NLL 3 2 6,5,5 3 238 + - 279NLL 3 4 6,6,6,5 11 239 + - 280 3 3 5,5,5 3 240 - - 232FTM 235FTM 237FTM

 0 No sperm FTM Failed to mate 1 Occasional sperm NLL No live litter - pr egnancy indicated by imp lantatio n sites 2 Continuous few sperm NLL* No live litter - pr egnancy indicated by late resorptions 3 Many scattered sperm 4 Solid masses of sperm ^ Estimated minimum cy cle length - cycle in progress at start or end of smearing period e Includes a period of extended oestrus (at least four consecutive days)266Huntingdon Life SciencesWAG0015

APPENDIX 16 Litter size - individual values - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropr opene Exposure level (ppm) : 0 198 505 2900 Group Animal Implantations Total litter size Live litter size on Day

number Day 1 1 5 10 1 241 15 15 15 15 15 242 15 14 14 14 14 243 14 14 14 14 14 244 17 16 16 16 16 245 16 13 13 12 1 2 246 18 17 17 17 16 247 15 14 14 14 14 248 15 15 15 15 15 249 17 16 15 15 15 250 17 16 16 16 16267Huntingdon Life SciencesWAG0015

APPENDIX 16 - continued Litter size - individual values - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Implantations Total litter size Live litter size on Day

number Day 1 1 5 10 2 251 14 14 14 14 14 252 13 12 11 9 9 253 12 12 12 11 11 254 16 14 14 14 14 255 19 14 13 7 7 256 14 14 13 12 12 257 NP 0 258 8 5 5 5 5 259 14 9 9 9 9 260 17 16 15 14 14 NP Not pregnant268Huntingdon Life SciencesWAG0015

APPENDIX 16 - continued Litter size - individual values - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Implantations Total litter size Live litter size on Day

number Day 1 1 5 10 3 261 16 13 12 9 8 262 W 10 8 263 TR 3 264 17 11 11 6 6 265 TLL 12 5 266 TLL 11 4 267 TLL 16 2 268 16 11 8 8 8 269 13 12 12 11 11 270 T LL 16 14 14 W Killed for welfare reasons TR Tota l resorption TLL Total litter loss269Huntingdon Life SciencesWAG0015

APPENDIX 16 - continued Litter size - individual values - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Implantations Total litter size Live litter size on Day

number Day 1 1 5 10 4 271 TR 6 272 FTM 0 273 TLL 12 3 1 274 TR 9 275 FTM 0 276 W 13 277 FTM 0 278 T R 8 279 W 9 280 NP 0 TR Total resorption FTM Failed to mate TLL Total litter loss W Killed for welfare reasons NP Not pregnant270Huntingdon Life SciencesWAG0015

APPENDIX 17 Offspring survival indices - individual values - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Post implantation Live birth Viability

number survival index index index

(%) (%) (%) 1 241 100 100 100

242 93 100 100 243 100 100 100 244 94 100 100 245 81 100 92 246 94 100 94 247 93 100 100 248 100 100 100 249 94 94 100 250 94 100 100271Huntingdon Life SciencesWAG0015

APPENDIX 17 - continued Offspring survival indices - individual values - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Post implantation Live birth Viability

numb er survival index index index

(%) (%) (%) 2 251 100 100 100

252 92 92 82 253 100 100 92 254 88 100 100 255 74 93 54 256 100 93 92 257 NP 258 63 100 100 259 64 100 100 260 94 94 93 NP Not pregnant272Huntingdon Life SciencesWAG0015

APPENDIX 17 - continued Offspring survival indices - individual values - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Ani mal Post implantation Live birth Viability

number survival index index index

(%) (%) (%) 3 261 81 92 67 262 W 80 0 263 TR 0 264 65 100 55 265 TLL 42 0 266 TLL 36 0 267 TLL 13 0 268 69 73 100 269 92 100 92 270 TLL 88 100 0 W Killed for welfare reasons TR Total resorption TLL Total litter loss273Huntingdon Life SciencesWAG0015

APPENDIX 17 - continued Offspring survival indices - individual values - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3, 3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Post implantation Live birth Viability

number survival index index index

(%) (%) (%) 4 271 TR 0 272 FTM 273 TLL 25 33 0 274 TR 0 275 FTM 276 W 0 277 FTM 278 TR 0 279 W 0 280 NP TR Total resorption FTM Failed to mate TLL Total litter loss W Killed for welfare reasons NP Not pregnant274Huntingdon Life SciencesWAG0015

APPENDIX 18 Sex ratio - individual values - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Total on Day Live on Day

number 1 1 5 10

M F %M M F %M M F %M M F %M 1 241 7 8 46.7 7 8 46.7 7 8 46.7 7 8 46.7

242 7 7 50.0 7 7 50.0 7 7 50.0 7 7 50.0 243 4 10 28.6 4 10 28.6 4 10 28.6 4 10 28.6 244 8 8 50.0 8 8 50.0 8 8 50.0 8 8 50.0 245 3 10 23.1 3 10 23.1 3 9 25.0 3 9 25.0 246 9 8 52.9 9 8 52.9 9 8 52.9 9 7 56.3 247 10 4 71.4 10 4 71.4 10 4 71.4 10 4 71.4 248 8 7 53.3 8 7 53.3 8 7 53.3 8 7 53.3 249 11 5 68.8 10 5 66.7 10 5 66.7 10 5 66.7 250 6 10 37.5 6 10 37.5 6 10 37.5 6 10 37.5275Huntingdon Life SciencesWAG0015

APPENDIX 18 - continued Sex ratio - individual values - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 290 0 Group Animal Total on Day Live on Day

number 1 1 5 10

M F %M M F %M M F %M M F %M 2 251 6 8 42.9 6 8 42.9 6 8 42.9 6 8 42.9

252 4 8 33.3 3 8 27.3 3 6 33.3 3 6 33.3 253 8 4 66.7 8 4 66.7 7 4 63.6 7 4 63.6 25 4 6 8 42.9 6 8 42.9 6 8 42.9 6 8 42.9 255 8 6 57.1 8 5 61.5 3 4 42.9 3 4 42.9 256 7 6 53.8 7 6 53.8 6 6 50.0 6 6 50.0 257 NP 258 2 3 40.0 2 3 40.0 2 3 40.0 2 3 40.0 259 5 4 55.6 5 4 55.6 5 4 55.6 5 4 55.6 260 11 5 68.8 10 5 66.7 10 4 71.4 10 4 71.4 NP Not pregnant276Huntingdon Life SciencesWAG0015

APPENDIX 18 - continued Sex ratio - individual values - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Total on Day Live on Day

number 1 1 5 10

M F %M M F %M M F %M M F %M 3 261 4 8 33.3 4 8 33.3 3 6 33.3 3 5 37.5 262 W 5 3 62.5 263 TR 264 7 4 63.6 7 4 63.6 2 4 33.3 2 4 33.3 265 TLL 3 2 60.0 266 TLL 3 1 75.0 267 TLL 1 0 100.0 268 4 4 50.0 4 4 50.0 4 4 50.0 4 4 50.0 269 8 4 66.7 8 4 66.7 7 4 63.6 7 4 63.6 27 0 TLL 6 8 42.9 6 8 42.9 W Killed for welfare reasons TR Total resorption TLL Total litter loss277Huntingdon Life SciencesWAG0015

APPENDIX 18 - continued Sex ratio - individual values - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - tr ifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Total on Day Live on Day

number 1 1 5 10

M F %M M F %M M F %M M F %M 4 271 TR 272 FTM 273 TLL 0 1 0.0 0 1 0.0 274 TR 275 FTM 276 W 277 FTM 278 TR 279 W 280 NP TR Total resorptio

n FTM Failed to mate TLL Total litter loss W Killed for welfare reasons NP Not pregnant278Huntingdon Life SciencesWAG0015

APPENDIX 19 Bodyweight - individual litter mean values for male offspring - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day of age number 1 5 10

Mean SD Mean SD Mean SD 1 241 6.6 0.2 10.5 0.4 16.1 0.8 242 6.7 0.4 10.7 0.5 16.7 1.2 24 3 7.4 0.1 11.7 0.4 18.3 0.8 244 6.9 0.3 10.9 0.8 18.3 2.0 245 6.5 0.2 11.1 0.1 18.7 0.1 246 6.9 0.4 11.1 1.0 18.6 1.4 247 6.8 0.3 10.6 0.6 17.2 1.3 248 6.8 0.3 10.7 0.3 16.9 0.8 249 7.1 0.6 11.7 1.0 18.2 2.0 250 6.5 0.5 9.9 0.9 16.1 1.0279Huntingdon Life SciencesWAG0015

APPENDIX 19 - continued Bodyweight - individual litter mean values for male offspring - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposur e level (ppm) : 0 198 505 2900 Group Animal Day of age number 1 5 10

Mean SD Mean SD Mean SD 2 251 7.3 0.5 11.1 0.7 16.7 0.5 252 7.3 0.3 11.1 0.6 16.5 0.7 253 7.6 0.3 11.4 0.3 17.5 0.6 254 7.2 0.3 9.8 0.9 15.5 1.4 255 7.0 0.4 10.6 1.4 18.1 2.8 256 6.6 0.6 10.5 0.7 15.6 0.9 257 NP 258 8.5 0.1 13.3 0.4 18.7 0.5 259 8.0 0.4 12.0 1.2 18.1 1.4 260 6.8 0.3 10.4 0.6 16.0 1.5 NP Not pregnant280Huntingdon Life SciencesWAG0015

APPENDIX 19 - continued Bodyweight - individual litter mean values for male offspring - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day of age number 1 5 10

Mean SD Mean SD Mean SD 3 261 7.3 0.4 11.3 0.4 17.2 0.5 262 W 263 TR 264 5.9 0.6 7.9 0.6 12.0 1.1 265 TLL 266 TLL 267 TLL 268 6.8 0.4 11.8 0.6 18.0 0.7 269 7.6 0.4 11.4 0.5 16.8 0.7 270 TLL 6.4 0.8 W Killed for welfare reasons TR Total resorption TLL Total litter loss281Huntingdon Life SciencesWAG0015

APPENDIX 19 - continued Bodyweight - individual litter mean values for female offspring - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluor opropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day of age number 1 5 10

Mean SD Mean SD Mean SD 1 241 6.1 0.3 9.8 0.7 15.4 1.1 242 6.2 0.4 10.4 0.5 16.0 0.8 2 43 6.6 0.2 10.5 0.6 16.8 0.5 244 6.5 0.4 10.2 1.2 17.1 2.5 245 6.2 0.3 10.4 0.6 17.6 0.7 246 6.2 0.4 10.7 0.9 18.0 1.3 247 6.5 0.2 10.4 0.2 17.1 0.3 248 6.4 0.1 10.2 0.4 15.6 0.9 249 7.2 0.3 11.6 0.5 18.3 1.2 250 6.0 0.5 9.5 0.9 15.3 1.7282Huntingdon Life SciencesWAG0015

APPENDIX 19 - continued Bodyweight - individual litter mean values for female offspring - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Expo sure level (ppm) : 0 198 505 2900 Group Animal Day of age number 1 5 10

Mean SD Mean SD Mean SD 2 251 6.7 0.5 10.2 0.7 15.8 0.9 252 7.2 0.4 11.4 0.8 16.6 1.0 253 6.7 0.5 10.1 1.2 15.5 2.2 254 7.1 0.4 10.0 0.7 15.3 0.9 255 6.8 0.4 11.1 0.9 18.0 2.5 256 6.2 0.3 9.8 0.7 14.7 0.9 257 NP 258 7.7 0.8 13.1 1.2 18.6 1.3 259 7.4 0.6 11.5 0.6 17.6 0.6 260 6.9 0.5 10.6 0.6 13.8 3.7 NP Not pregnant283Huntingdon Life SciencesWAG0015

APPENDIX 19 - continued Bodyweight - individual litter mean values for female offspring - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day of age number 1 5 1 0

Mean SD Mean SD Mean SD 3 261 6.3 0.7 10.0 1.6 16.1 2.2 262 W 263 TR 264 6.0 0.3 7.5 0.4 11.6 0.2 265 TLL 266 TLL 267 TLL 268 6.2 0.4 11.0 1.3 17.5 1.9 269 6.5 0.6 10.2 0.9 15.1 1.1 270 TLL 6.3 0.3 W Killed for welfare reasons TR Total resorption TLL Total litter loss284Huntingdon Life SciencesWAG0015

APPENDIX 19 - continued Bodyweight - individual litter mean values for female offspring - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Day of age number 1

Mean SD 4 271 TR 272 FTM 273 TLL 5.5 - 274 TR 275 FTM 276 W 277 FTM 278 TR 279 W 280 NP TR Total resorption FTM Failed to mate TLL Total litter loss W Killed for welfare reasons NP Not pregnant285Huntingdon Life SciencesWAG0015

APPENDIX 20 Clinical signs - individual observations for offspring - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Dam

number Number/Sex

of offspring Observation Day of age 1 241 7M & 8F Cold to touch 1 250 1F Pale 1 2 251 6M & 8F Cold to touch 1 252 1 F Darker in colour than other pups 1 Found dead 2 253 1M Bruising on head 1

1M Bruising on hindlimb, Right 1 254 1M Bruising on head 1 Small amount of milk seen in abdomen 1 1M Bruising on head 1 255 3M & 4F Cold to touch 1

4M Cold to touch 1

Pups missing 2 1F Cold to touch 1

Pup missing 2 1M Cold to touch 1

Found dead 2 Only animals with observations are presented286Huntingdon Life SciencesWAG0015

APPENDIX 20 - continued Clinical signs - individual observations for offspring - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 19 8 505 2900 Group Dam

number Number/Sex

of offspring Observation Day of age 2 256 1F Bruising on head 1 260 1F Pale 1 Cold to touch 1 Pup missing 3 3 261 3 M & 5 F Dark in colour 1 Un deractive 1 Cold to touch 1 - 2 Sl ightly cold to touch 3 1F Dark in colour 1

Un deractive 1 Cold to touch 1 Pup missing 2 1M & 1F Dark in colour 1

Un deractive 1 Cold to touch 1 - 2 Pups missing 3 1F Dark in colour 1

Un deractive 1 Cold to touch 1 - 2 Slightly cold to touch 3 Pup missing 9 Only animals with observations are presented287Huntingdon Life SciencesWAG0015

APPENDIX 20 - continued Clinical signs - individual observations for offspring - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trif luoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Dam

number Number/Sex

of offspring Observation Day of age 3 262 W 3F Cold to touch 1 No milk present in stomach 1 Dark in colour 1 Un deractive 1 Dam and pups killed for welfare reasons 1 264 2M & 4F Cold to touch 2

2M Cold to touch 2

Pups missing 3 266 TLL 1M Cold to touch 1 Dark in colour 1 Inactive 1 Killed for welfare reasons 1 267 TLL 1M No food present in stomach 1 Dark in colour 1 Un deractive 1 Killed for welfare reasons 1 Only animals with observations are presented W Killed for welfare reasons TLL Total litter loss288Huntingdon Life SciencesWAG0015

APPENDIX 20 - continued C linical signs - individual observations for offspring - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Dam

number Number/Sex

of offspring Observation Day of age 3 269 7M & 4F Cold to touch 1 Milk present in stomach 1 1M Cold to touch 1

Milk present in stomach 1 Found dead 2 270 TLL 3M & 3F Cold to touch 1 Pups missing 2 1M C old to touch 1

Pale 1 Pup missing 2 1M & 2F Cold to touch 1

Found dead 2 1M & 3F Cold to touch 1

Dark, underactive , little milk present in stomach 2 Killed for welfare reasons 2 Only animals with observations are presented TLL Total litter loss289Huntingdon Life SciencesWAG0015

APPENDIX 21 Macropathology - individual findings for offspring killed or dying before scheduled termination - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group F0 Female Number Death

Code Day

of age Sex of offspring Macroscopic observation 1 245 F 5 F Milk in stomach 246 F 6 F No milk in stomach 2 252 F <1 M M ilk in stomach

F 2 F Liver, Mottled, Pale No milk in stomach 255 F <1 F No milk in stomach

F 2 M No milk in stomach290Huntingdon Life SciencesWAG0015

APPENDIX 21 - continued Macropathology - individual findings for offspring dying before scheduled termination (F1) Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group F0 Female Number Death

Code Day

of age Sex of offspring Macrosc opic observation 3 261 F <1 F No milk in stomach

F 1 F Milk in stomach 262 W <1 M No milk in stomach

W <1 F Milk in stomach F <1 3 x M: 2 x F No milk in stomach F <1 M A small amount of milk in stomach 265 F <1 3 x M: 2 x F No milk in stomach 266 W <1 M No milk in stomach

F <1 2 x M No milk in stomach F <1 F Milk in stomach 267 W <1 M No milk in stomach 269 F 2 M Milk in stomach 270 F 2 1 x M: 2 x F No milk in stomach

W 2 1 x M: 3 x F No milk in stomach Only animals with abnormalities presented291Huntingdon Life SciencesWAG0015

APPENDIX 22 Macropathology - individual findings for offspring killed at scheduled termination - F1 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group F0 Female Number Sex of

offspring Macroscopic observation 2 254 F Left forepaw, third digit, pale swelling underlying tattoo, 4 mm Only animals with abnormalities presented292Huntingdon Life SciencesWAG0015

APPENDIX 23 Sperm analysis - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Motile Progressively ----- Cauda epididymis ----- -------- Testis -------- number number sperm motile sperm Weight Sperm count Total Weight Sperm count Total (%) (%) (g) (millio ns/g) (millio

n) (g) (millio ns/g) (millio

n) 1 201 94 50 0.236 1169 276 2.00 202 404 202 97 26 0.255 333 85 1.96 199 391 B 203 97 65 0.238 1237 294 1.61 220 353 204 95 79 0.254 955 243 1.71 196 335 205 91 64 0.252 1203 303 1.87 115 216 206 92 68 0.2 82 1526 430 1.93 241 466 207 89 64 0.183 1196 219 1.82 175 318 208 99 53 0.236 1080 255 1.73 249 430 209 97 74 0.247 1032 255 1.65 187 309 210 90 51 0.218 1169 255 1.75 208 364 B Excluded from group mean cauda epididymal count values, suspected sample preparation/dilution error293Huntingdon Life SciencesWAG0015

APPENDIX 23 - continued Sperm analysis - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Motile Progressively ----- Cauda epididymis ----- -------- Testis -------- number number sperm motile sperm Weight Sperm count Total Weight Sperm count Total (%) (%) (g) (millio ns/g) (mi llio

n) (g) (millio ns/g) (millio

n) 2 211 91 43 0.222 1081 240 1.76 149 262 212 95 54 0.236 1377 325 1.74 147 255 213 99 22 0.248 877 218 1.91 294 561 214 95 55 0.224 1256 281 1.94 281 545 215 92 75 0.219 1490 326 1.89 251 474 216 99 19 0.264 1056 279 1.87 210 393 217 99 41 0.245 963 236 1.78 175 311 218 86 43 0.252 1133 286 1.68 125 211 219 100 75 0.219 1173 257 1.85 107 197 220 95 48 0.237 1041 247 2.00 208 416294Huntingdon Life SciencesWAG0015

APPENDIX 23 - continued Sperm analysis - indivi dual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Motile Progressively ----- Cauda epididym is ----- -------- Testis -------- number number sperm motile sperm Weight Sperm count Total Weight Sperm count Total (%) (%) (g) (millio ns/g) (millio

n) (g) (millio ns/g) (millio

n) 3 221 99 11 0.229 1104 253 1.82 115 209 222 94 60 0.245 1387 340 2.12 268 568 223 91 66 0.22 955 210 1.57 159 250 224 99 55 0.233 986 230 1.81 164 297 225 96 50 0.225 1405 316 1.83 218 399 226 98 36 0.231 1192 275 1.77 108 190 227 95 42 0.248 896 222 1.90 187 355 228 63 10 0.237 642 152 1.73 181 313 229 97 66 0.233 968 226 1.72 66 114 230 0 0 0.091 37 3 0.31 11 3 A A Few sperm in sample, excluded from group mean motility values295Huntingdon Life SciencesWAG0015

APPENDIX 23 - continued Sperm analysis - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Motile Progressively ----- Cauda epididymis ----- -------- Testis -------- nu mber number sperm motile sperm Weight Sperm count Total Weight Sperm count Total

(%) (%) (g) (millio ns/g) (millio

n) (g) (millio ns/g) (millio

n) 4 231 96 55 0.243 624 152 1.79 235 420

232 84 57 0.171 803 137 1.85 248 459 233 90 25 0.225 1036 233 1.65 130 214 234 94 54 0.214 931 199 1.72 209 360 235 96 47 0.174 1273 222 1.78 177 314 236 94 42 0.177 937 166 1.74 111 194 237 88 37 0.181 1299 235 1.59 177 282 238 90 37 0.219 900 197 1.72 182 313 239 97 31 0.170 1084 184 1.63 148 241 240 94 41 0.215 1148 247 1.95 168 328296Huntingdon Life SciencesWAG0015

APPENDIX 24 Sperm motion data - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal VAP VSL VCL ALH BCF STR LIN Elongation Area Rapid Medium Slow Static number number (um/s) (um/s) (um/s) (um) (Hz) (%) (%) (%) (um sq) (%) (%) (%) (%) 1 201 138 94 287 23 26 69 34 28 439 62 3 29 6 202 124 85 260 25 28 69 35 33 438 32 7 59 3 203 138 107 289 21 25 77 38 20 399 71 3 23 3 204 144 110 303 21 22 77 37 21 492 85 0 9 5 205 124 96 267 20 22 77 36 18 442 71 5 14 9 206 143 103 323 23 24 72 32 23 444 76 2 14 8 207 137 107 303 21 25 78 36 20 477 68 2 20 11 208 142 98 309 24 29 69 33 31 730 64 5 30 1 209 151 111 339 23 25 74 33 22 557 84 2 10 3 210 138 108 313 23 26 77 35 23 448 55 2 33 10297Huntingdon Life SciencesWAG0015

APPENDIX 24 - continued Sperm motion data - indivi dual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal VAP VSL VCL ALH BCF STR LIN Elongation Area Rapid Medium Slow Static number number (um/s) (um/s) (um/s) (um) (Hz) (%) (%) (%) (um sq) (%) (%) (%) (%) 2 211 116 85 238 21 27 73 38 20 406 49 4 39 9 212 143 100 291 22 25 70 35 27 553 63 2 30 5 213 120 83 268 24 29 69 33 29 469 27 6 65 1 214 145 104 297 23 27 72 36 24 481 64 5 27 5 215 123 99 264 19 24 80 38 17 388 77 5 10 8 216 110 79 229 24 30 72 37 32 661 24 7 67 1 217 132 99 273 23 27 75 39 25 420 47 6 46 1 218 120 84 260 21 23 70 33 23 435 51 7 28 14 219 137 106 284 21 24 78 38 23 473 82 2 16 0 220 113 83 245 20 29 73 36 27 777 56 9 30 5298Huntingdon Life SciencesWAG0015

APPENDIX 24 - continued Sperm motion data - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal VAP VSL VCL ALH BCF STR LIN Elongation Area Rapid Medium Slow Static number number (um/s) (um/s) (um/s) (um) (Hz) (%) (%) (%) (um sq) (%) (%) (%) (%) 3 221 106 74 215 22 32 70 37 27 442 14 3 82 1 222 133 102 273 19 27 76 39 26 521 68 5 21 6 223 127 87 253 22 21 69 35 24 413 78 2 11 9 224 132 98 272 22 28 73 37 27 482 62 6 32 1 225 115 76 224 19 22 68 36 21 349 65 5 26 4 226 124 86 239 22 27 70 39 27 478 43 6 48 2 227 125 91 247 23 25 73 38 26 502 44 5 45 5 228 90 59 182 19 24 68 35 23 275 13 9 41 37 229 125 93 258 19 23 75 38 22 543 77 5 15 3 230 0 0 0 0 0 0 0 0 0 0 0 0 100 A A Few sperm in sample, excluded from group mean values299Huntingdon Life SciencesWAG0015

APPENDIX 24 - continued Sperm motion data - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal VAP VSL VCL ALH BCF STR LIN Elongation Area Rapid Medium Slow Static number number (um/s) (um/s) (um/s) (um) (Hz) (%) (%) (%) (um sq) (%) (%) (%) (%) 4 231 146 102 320 22 27 70 34 27 780 71 3 23 4 232 143 115 290 20 28 80 41 19 374 60 4 21 16 233 102 69 206 22 29 69 36 30 534 30 9 50 10 234 128 95 258 21 28 74 38 24 415 63 6 25 6 235 124 82 257 21 28 68 34 30 787 61 6 28 4 236 114 87 226 20 28 76 40 23 364 47 9 38 6 237 124 86 265 21 28 71 35 27 551 43 5 40 12 238 123 86 259 22 29 70 35 22 435 45 6 39 10 239 114 80 247 22 31 71 36 29 578 40 11 46 3 240 130 95 271 24 29 73 36 27 429 46 7 42 6300Huntingdon Life SciencesWAG0015

APPENDIX 25 Sperm mo

rphology - individ

ual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Number of Normal Total ab normal Decapitate Head ab normal Detached/broken Other number number sperm neck/midpiece/tail abnormality

examined number % number % number % number % number % number % 1 201 200 197 98.5 3 1.5 2 1.0 1 0.5 0 0.0 0 0.0 202 200 199 99.5 1 0.5 0 0.0 0 0.0 0 0.0 1 0.5 203 200 197 98.5 3 1.5 1 0.5 2 1.0 0 0.0 0 0.0 204 200 197 98.5 3 1.5 1 0.5 2 1.0 0 0.0 0 0.0 205 200 196 98.0 4 2.0 1 0.5 1 0.5 2 1.0 0 0.0 206 200 199 99.5 1 0.5 0 0.0 0 0.0 0 0.0 1 0.5 207 200 198 99.0 2 1.0 1 0.5 1 0.5 0 0.0 0 0.0 208 200 193 96.5 7 3.5 7 3.5 0 0.0 0 0.0 0 0.0 209 200 199 99.5 1 0.5 0 0.0 1 0.5 0 0.0 0 0.0 210 200 196 98.0 4 2.0 1 0.5 2 1.0 0 0.0 2 1.0301Huntingdon Life SciencesWAG0015

APPENDIX 25 - continued Sperm mo

rphology - individual values - F0 generatio n Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Number of Normal Total ab normal Decapitate Head ab normal Detached/broken Other number number sperm neck/midpiece/tail abnormality

examined number % number % number % number % number % number % 2 211 200 186 93.0 14 7.0 2 1.0 11 5.5 0 0.0 1 0.5 212 200 198 99.0 2 1.0 1 0.5 1 0.5 0 0.0 0 0.0 213 200 197 98.5 3 1.5 2 1.0 1 0.5 0 0.0 0 0.0 214 200 198 99.0 2 1.0 0 0.0 2 1.0 0 0.0 0 0.0 215 200 194 97.0 6 3.0 4 2.0 1 0.5 1 0.5 0 0.0 216 200 195 97.5 5 2.5 1 0.5 2 1.0 1 0.5 1 0.5 217 200 188 94.0 12 6.0 10 5.0 0 0.0 2 1.0 0 0.0 218 200 196 98.0 4 2.0 1 0.5 1 0.5 1 0.5 1 0.5 219 200 192 96.0 8 4.0 4 2.0 3 1.5 1 0.5 1 0.5 220 213 200 93.9 13 6.1 5 2.3 8 3.8 0 0.0 0 0.0302Huntingdon Life SciencesWAG0015

APPENDIX 25 - continued Sperm mo

rphology - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Number of Normal Total ab normal Decapitate Head ab normal Detached/broken Other number number sperm neck/midpiece/tail abnormality

examined number % number % number % number % number % number % 3 221 200 195 97.5 5 2.5 4 2.0 1 0.5 0 0.0 0 0.0 222 200 197 98.5 3 1.5 0 0.0 2 1.0 2 1.0 0 0.0 223 200 186 93.0 14 7.0 12 6.0 1 0.5 2 1.0 0 0.0 224 200 191 95.5 9 4.5 6 3.0 3 1.5 0 0.0 0 0.0 225 200 193 96.5 7 3.5 4 2.0 3 1.5 0 0.0 0 0.0 226 200 187 93.5 13 6.5 10 5.0 1 0.5 2 1.0 0 0.0 227 10 8 80.0 2 20.0 2 20.0 0 0.0 0 0.0 0 0.0 A 228 200 194 97.0 6 3.0 2 1.0 1 0.5 3 1.5 0 0.0 229 200 190 95.0 10 5.0 0 0.0 9 4.5 1 0.5 1 0.5 230 0 A A Few / no sperm in sample, excluded from group mean values303Huntingdon Life SciencesWAG0015

APPENDIX 25 - continued Sperm mo

rphology - individual valu es - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group Animal Number of Normal Total ab normal Decapitate Head ab normal Detached/broken Other number number sperm neck/midpiece/tail abnormality

examined number % number % number % number % number % number % 4 231 200 189 94.5 11 5.5 4 2.0 3 1.5 3 1.5 1 0.5 232 200 160 80.0 40 20.0 15 7.5 18 9.0 8 4.0 2 1.0 233 200 161 80.5 39 19.5 31 15.5 5 2.5 5 2.5 1 0.5 234 200 186 93.0 14 7.0 8 4.0 6 3.0 1 0.5 1 0.5 235 200 177 88.5 23 11.5 15 7.5 3 1.5 3 1.5 3 1.5 236 200 191 95.5 9 4.5 5 2.5 3 1.5 2 1.0 1 0.5 237 200 191 95.5 9 4.5 3 1.5 4 2.0 2 1.0 1 0.5 238 200 184 92.0 16 8.0 5 2.5 11 5.5 1 0.5 1 0.5 239 200 173 86.5 27 13.5 15 7.5 10 5.0 2 1.0 1 0.5 240 200 193 96.5 7 3.5 2 1.0 5 2.5 0 0.0 0 0.0304Huntingdon Life SciencesWAG0015

APPENDIX 26 Request ID: 284410 Organ weights - individual absolute values (g) for males - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group /Sex Animal Number Terminal Bodyw eight Lt.

Epididymis Rt.

Epididymis Pituitary Prostate Seminal Vesicles Lt. Testis Rt. Testis 1M 201 533 0.667 0.648 0.014 1.538 1.405 2.00 1.99 202 485 0.759 0.699 0.013 1.428 1.376 1.96 1.86 203 518 0.618 0.550 0.015 1.261 1.180 1.61 1.69 204 561 0.689 0.705 0.016 1.510 2.251 1.71 1.68 205 582 0.731 0.667 0.020 1.493 1.523 1.87 1.85 206 589 0.799 0.745 0.015 1.233 2.418 1.93 1.96 207 537 0.592 0.638 0.012 1.393 1.338 1.82 1.88 208 542 0.641 0.668 0.016 1.453 1.890 1.73 1.68 209 564 0.705 0.666 0.016 1.335 1.588 1.65 1.63 210 518 0.603 0.580 0.012 0.998 1.697 1.75 1.70 2M 211 503 0.585 0.584 0.011 0.964 1.692 1.76 1.77 212 407 0.700 0.651 0.009 1.022 1.910 1.74 1.74 213 511 0.648 0.588 0.012 1.178 1.164 1.91 1.8 2 214 564 0.642 0.665 0.012 1.028 1.254 1.94 1.94 215 515 0.651 0.629 0.011 1.085 1.517 1.89 1.96 216 553 0.683 0.597 0.014 0.999 1.356 1.87 1.79 217 471 0.686 0.592 0.008 0.968 0.849 1.78 1.77 218 437 0.636 0.604 0.010 0.891 1.554 1.68 1.67 21 9 439 0.636 0.626 0.009 0.972 1.279 1.85 1.91 220 436 0.650 0.637 0.011 0.975 0.996 2.00 1.98305Huntingdon Life SciencesWAG0015

APPENDIX 26 - continued Request ID: 284410 Organ weights - individual absolute values (g) for males - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group /Sex Animal Number Terminal Bodyweight Lt.

Epididymis Rt.

Epididymis Pituitary Prostate Seminal Vesicles Lt. Testis Rt. Testis 3M 221 465 0.604 0.554 0.010 0.726 1.434 1.82 1.82 222 410 0.671 0.641 0.010 0.712 1.174 2.12 2.15 223 425 0.578 0.529 0.013 0.687 1.236 1.57 1.57 224 438 0.687 0.617 0.010 0.821 1.191 1.81 1.84 225 470 0 .631 0.656 0.013 1.160 1.259 1.83 1.94 226 441 0.652 0.644 0.011 1.460 0.959 1.77 1.75 227 511 0.677 0.692 0.012 0.891 0.883 1.90 1.89 228 452 0.627 0.591 0.011 0.855 0.925 1.73 1.69 229 446 0.631 0.605 0.009 0.894 1.187 1.72 1.65 230 469 0.207 0 .652 0.009 1.167 1.034 0.31 2.13 4M 231 469 0.618 0.585 0.011 0.463 1.601 1.79 1.79 232 409 0.532 0.636 0.012 0.264 0.899 1.85 1.78 233 380 0.602 0.565 0.009 0.627 1.320 1.65 1.64 234 414 0.595 0.595 0.008 0.636 0.925 1.72 1.68 235 452 0.595 0.623 0.008 0.781 0.924 1.78 1.77 236 453 0.559 0.552 0.006 0.843 1.253 1.74 1.72 237 403 0.582 0.590 0.011 0.643 1.139 1.59 1.56 238 387 0.573 0.547 0.009 0.540 1.468 1.72 1.65 239 444 0.494 0.559 0.009 0.730 0.963 1.63 1.58 240 423 0.620 0.620 0.011 0.722 0.759 1.95 1.99306Huntingdon Life SciencesWAG0015

APPENDIX 27 Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluor opropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 201 Sex: Male Dose Group: 1 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 532.9 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Pr ostate, Rt. Epididymis, Rt. Testis307Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and hist opathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 202 Sex: Male Dose Group: 1 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 484.9 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Pr ostate, Rt. Epididymis, Rt. Testis308Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluor opropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 203 Sex: Male Dose Group: 1 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 5 1 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 518.0 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** Prostate Interstitial Inflammation, Minimal The following examined tissues had no gross observations and no findings microscopically: Rt. Epididymis, Rt. Testis309Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 2 89382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 204 Sex: Male Dose Group: 1 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 560.5 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had n o gross observations and no findings microscopically: Pr ostate, Rt. Epididymis, Rt. Testis310Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 205 Sex: Male Dose Group: 1 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of D eath: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 581.7 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Pr ostate, Rt. Epididymis, Rt. Testis311Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure l evel (ppm) : 0 198 505 2900 Animal Number: 206 Sex: Male Dose Group: 1 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 589.0 g PATHOLOGY OBSERVATION S MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** Prostate Acinar Cell Atrophy, Focal, Minimal The following examined tissues had no gross observations and no findings microscopically: Rt. Epididymis, Rt. Testis312Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 207 Sex: Male Dose Group: 1 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 537.4 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Pr ostate, Rt. Epididy

mis, Rt. Testis313Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3, 3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 208 Sex: Male Dose Group: 1 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight : 541.8 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** Prostate Inter stitial Inflammation, Slight The following examined tissues had no gross observations and no findings microscopically: Rt. Epididymis, Rt. Testis314Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individ ual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 209 Sex: Male Dose Group: 1 Phase : Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 563.6 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Pr ostate, Rt. Epididymis, Rt. Testis315Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3 ,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 210 Sex: Male Dose Group: 1 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 S ubgroup: 1 Terminal Bodyweight: 517.7 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Teeth Incisor(s) pale, Lower, Right The following examined tissues had no gross observations and no findings microscopically: Pr ostate, Rt. Epididymis, Rt. Testis316Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 ge neration Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 211 Sex: Male Dose Group: 2 Phase: Treatment Sacrific e Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 502.5 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Pr ostate317Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 212 Sex: Male Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 407.0 g PATHOLO GY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Spleen Spleen Capsule thickened, Multiple, areas. Capsular/Subcapsular Inflammatio n, Slight Adhesions/Inflammation/Fibrosis, Minimal The following examined tissues had no gross observations and no findings microscopically: Pr ostate318Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 213 Sex: Male D ose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 510.5 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** Prostate Interstitial Inflammation, Focal, Minimal The following examined tissues had no gross observations and no findings microscopically:

-319Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3 ,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 214 Sex: Male Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 564.1 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Interstitial Inflammation , Minimal Spleen Spleen Capsule thickened, Posterior pole, A few, areas, 1mm. Capsular/Subcapsular Inflammation, Minimal Adhesions/Inflammation/Fibrosis, Minimal The following examined tissues had no gross observationsand no findings microscopically:

-320Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 215 Sex: Male Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Sub group: 1 Terminal Bodyweight: 514.5 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Spleen Spleen Capsule thickened, areas. Capsular/Subcapsular Inflammation, Slight Adhesions, Anterior pole, capsule to adipose tissue. Capsular Thickening, Minimal Adhesions/Inflammation/Fibrosis, Slight The following examined tissues had no gross observations and no findings microscopically: Pr ostate321Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 216 Sex: Male Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 553.1 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Skin Hair loss, Side(s) of face, diffuse. Spleen Spleen Capsule thickened, Multiple, areas. Capsular/Subcapsular Inflammation, Slight Adhesions, capsule to adipose tissue. Capsular Thickening, Minimal Adhesions/Inflammation/Fibrosi s, Slight The following examined tissues had no gross observations and no findings microscopically: Pr ostate322Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 217 Sex: Male Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 471.2 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Spleen Spleen Capsule thickened, Multiple, areas. Capsular/Subcapsular Inflammation, Minimal Adhesions/Inflammation/Fibrosis, Minimal The following examined tissues had no gross observations a nd no findings microscopically: Pr ostate323Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3, 3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 218 Sex: Male Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 436.6 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Prostate Small No significant lesions Spleen Spleen Capsule thickened, Multiple, areas. Capsular/Subcapsular Inflammation, Minimal Capsular Thickening, Minimal The following examined tissues had no gross observations and no findings micr oscopically:

-324Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 219 Sex: Male Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyw eight: 438.7 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Spleen Spleen Capsule thickened, areas. Capsular/Subcap sular Inflammation, Minimal Capsular Thickening, Minimal The following examined tissues had no gross observations and no findings microscopically: Pr ostate325Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 220 Sex: Male Dose Group: 2 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 435.5 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Spleen Spleen Capsule thickened, Anterior pole, A few, areas. Capsular/Subcapsular Inflammation, Focal, Minimal The following examined tissues ha d no gross observations and no findings microscopically: Pr ostate326Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - tr ifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 221 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of De ath: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 464.6 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Prostate Small No significant lesions The following examined tissues had no gross observations and no findings microscopically:

-327Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 222 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 410.2 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Prostate Small No significant lesions Spleen Spleen Capsule thickened, Punctate, A few, areas. Capsular/Subcapsular Inflammation, Focal, Minimal Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically:

-328Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 ge neration Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 223 Sex: Male Dose Group: 3 Phase: Treatment Sacrific e Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 425.4 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Prostate Small Interstitial Inflammation, Minimal Acinar Cell Atrophy, Focal, Slight Skin Skin Hair loss, Side(s) of face, Right Scab(s), Present Scab(s), Side(s) of face, Right, One, 1mm. Epidermal Hyperplasia, Minimal The following examined tissues had no gross observations and no findings microscopically:

-329Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropatho logy and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 22 4 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 437.8 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Prostate Small Abscessation, Focal, Minimal Skin Hair loss, Side(s) of face, Right, diffuse. Spleen Spleen Capsule thickened, Anterior pole, an area. Capsular/Subcapsular Inflammation, Minimal Adhesions, Anterior pole, capsule to adipose tissue. Capsular Thickening, Minimal Adhesions/Inflammation/Fibrosis, Minimal T eeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically:

-330Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 225 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 470.2 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Interstitial Inflammation, Minimal Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically:

-331Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoroprop ene Exposure level (ppm) : 0 198 505 2900 Animal Number: 226 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 440.5 g PATHOL OGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Spleen Spleen Adhesions, Multiple, diffuse. Capsular/Subcapsular Inflammation , Slight Capsular Thickening, Slight Adhesions/Inflammation/Fibrosis, Slight The following examined tissues had no gross observations and no findings microscopically: Pr ostate332Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 227 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 511.2 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Prostate Small Interstitial Inflammation, Slight Spleen Spleen Capsule thickened, A few, areas. Capsular/Subcapsular Inflammation, Slight Adhesions, Anterior pole, capsule to adipose tissue. Capsular Thickening, Minimal The following examined tissues had no gross observations and no findings microscopically:

-333Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropen e Exposure level (ppm) : 0 198 505 2900 Animal Number: 228 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 451.8 g PATHOLOG Y OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Prostate Small No significant lesions Spleen Spleen Capsul e thickened Capsular/Subcapsular Inflammation, Moderate Capsular Thickening, Slight Adhesions/Inflammation/Fibrosis, Moderate Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically:

-334Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 229 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 445.8 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Prostate Small Ac inar Cell Atrophy, Focal, Slight Spleen Spleen Capsule thickened, Anterior pole, areas. Capsular Thickening, Focal, Minimal Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically:

-335Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trif luoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 230 Sex: Male Dose Group: 3 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup : 1 Terminal Bodyweight: 468.6 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Lt. Epididymis Lt. Epididymis Small No significant lesions Spleen Spleen Capsule thickened, Multiple, areas. Capsular/Subcapsular Inflammation, Minimal Capsular Thickening, Minimal Adhesions/Inflammation/Fibrosis, Minimal Lt. Testis Lt. Testis Small No significant lesions Pr ominent tubules, A few Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically: Pr ostate336Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 231 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 468.5 g PATHOLOGY OBSERVATIONS MAC ROPATHOLOGY HI STOPATHOLOGY Prostate Prostate Small No significant lesions Skin Hair loss, cranial region and si des of face, diffuse. Spleen Spleen Capsule thickened, Anterior pole, areas. Capsular/Subcapsular Inflammation, Focal, Minimal Teeth Incisor(s) pale, Lower The following examined tissues had no gross observation s and no findings microscopically: Rt. Epididymis, Rt. Testis337Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluor opropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 232 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 409.3 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Prostate Small No significant lesions Spleen Spleen Capsule thickened Capsular/Subcapsular Inflammation, Slight Adhesions, to adipose tissue. Capsular Thickening, Moderate Adhesions/Inflammation/Fibrosis, Moderate Rt. Testis Spermatocytes Degeneration, Minimal Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically: Rt. Epididymis338Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 233 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 379.5 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HIS TOPATHOLOGY Prostate Prostate Small No significant lesions Skin Hair loss, cranial region extending to dorsal cervical region and sides of face. Spleen Spleen Capsule thickened, Anterior pole, two areas. No significant lesions Teeth Incisor(s) pale, Lower The following examined tissues had no gross observati ons and no findings microscopically: Rt. Epididymis, Rt. Testis339Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trif luoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 234 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Deat h: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 414.2 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Prostate Small Interstitial Inflammation, Slight Skin Hair loss, cranial region and ventral cervical region. Spleen Spleen Capsule thickened, areas, up to 1mm. Capsular Thickening, Focal, Minimal Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically: Rt. Epididymis, Rt. Testis340Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual f indings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 235 Sex: Male Dose Group: 4 Phase: Tre atment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 452.2 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Prostate Small No significant lesions Spleen Spleen Capsule thickened Capsular/Subcapsular Inflammation, Moderate Adhesions, capsulat to adipose tissue. Capsular Thickening, Slight Adhesions/Inflammation/Fibrosis, Slight Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically: Rt. Epididymis, Rt. Testis341Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - brom o - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 236 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodywe ight: 453.1 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Prostate Small Interstitial Inflammation, Minim al Spleen Spleen Capsule thickened, a few areas. Capsular/Subcapsular Inflammation, Minimal Adhesions, Anterior pole, capsule to adipose tissue. Capsular Thickening, Slight Adhesions/Inflammation/Fibrosis, Min imal Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically: Rt. Epididymis, Rt. Testis342Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 237 Sex: M ale Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 403.2 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Prostate Small No significant lesions Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically: Rt. Epididymis, Rt. Testis343Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - tri fluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 238 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 386.6 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Prostate Small Abscessation, Focal, Slight Spleen Spleen Capsule thickened Capsular/Subcapsular Inflammation, Moderate Adhesions, capsule to abdominal wall. Capsular Thickening, Slight Adhesions/Inflammation/Fibrosis, Slight Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically: Rt. Epididymis, Rt. Testis344Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macr opathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 239 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 444.0 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Prostate Prostate Small Interstitial Inflammation, Focal, Minimal Skin Hair loss, Cranial region, diffuse. Spleen S pleen Capsule thickened, Multiple, areas. Capsular/Subcapsular Inflammation, Slight Capsular Thickening, Slight Adhesions/Inflammation/Fibrosis, Slight Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically: Rt. Epididymis, Rt. Testis345Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 240 Sex: Male Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifi ce Date of Death: 07 - Sep - 12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 422.6 g PATHOLOGY OBSERVATIONS MACROPATHOLOGY HI STOPATHOLOGY Prostate Prostate Small No significant lesions Spleen Spleen Capsule thickened, Multiple, areas. Capsular/Subcapsular Inflammation, Minimal Capsular Thickening, Minimal Adhesions/Inflammation/Fibrosis, Minimal Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically: Rt. Epididymis, Rt. Testis346Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Anima l Number: 241 Sex: Female Dose Group: 1 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 05 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 339.8 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no g ro ss observations and no findings microscopically: Ovaries347Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 242 Sex: Female Dose Group: 1 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 05 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 310.0 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPI C FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Ovaries348Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and hi stopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 243 Sex: Fem ale Dose Group: 1 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 03 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 318.7 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Ovaries349Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

tr ifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 244 Sex: Female Dose Group: 1 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 05 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 380.3 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** * **ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Ovaries350Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individua l findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 245 Sex: Female Dose Group: 1 Phase : Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 05 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 336.3 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Ovaries351Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3 ,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 246 Sex: Female Dose Group: 1 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodywei ght: 378.7 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCO PIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Ovaries352Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generati on Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 247 Sex: Female Dose Group: 1 Phase: Lactatio n Sacrifice St atus: Final phase sacrifice Date of Death: 06 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 306.8 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Ovaries353Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene E xposure level (ppm) : 0 198 505 2900 Animal Number: 248 Sex: Female Dose Group: 1 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 06 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 349.7 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Skin Hair loss, Forelimb(s) The following examined tissues had no gross observati ons and no findings microscopically: Ovaries354Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - brom o - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 249 Sex: Female Dose Group: 1 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 06 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 340.9 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS REC ORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Ovaries355Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 250 Sex: Female Dose Group : 1 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 06 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 354.6 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Skin Hair loss, Forelimb(s) The following examined tissues had no gross observations and no findings microscopically: Ovaries356Huntingdon Life SciencesWAG0015

APPENDIX 27 - continue d Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 251 Sex: Female Dose Group: 2 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 06 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 323.5 g

PATHOLOGY OBSERVATIONS MACROPATHOL OGY HISTOPATHO LOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Ovaries357Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - bromo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 252 Sex: Female Dose Group: 2 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 04 - S ep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 292.3 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Ovaries358Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 M acropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 253 Sex: Female Dose Group: 2 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 05 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 300.9 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Ovaries359Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoroprop ene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 254 Sex: Female Dose Group: 2 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 05 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 370.5 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Ovaries360Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histop athology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 255 Sex: Female Dose Group: 2 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 13 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 339.8 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Skin Hair loss, Ventral surface, Patchy The following examined tissues had no gross observations and no findings microscopically: Ovaries361Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exp osure level (ppm) : 0 198 505 2900 Animal Number: 256 Sex: Female Dose Group: 2 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 05 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 331.2 g

PATHOLOGY OB SERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Kidneys Kidneys Pelvic dilatation Cortical Tubular Basophilia, Bilateral, Slight Cortical Tubular Dilatation, Unilateral, Minimal Pelvic Dilatation, Present The following examined tissues had no gross observations and no findings microscopically: Ovaries362Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 2893 82 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Ani mal Number: 257 Sex: Female Dose Group: 2 Phase: Gestation Sacrifice Status: Final phase sacrifice Date of Death: 29 - Aug - 12 Day of Death: 25 Week of Death: 4 Subgroup: 1 Terminal Bodyweight: 285.4 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** Ovaries Reduced Size of Corpora Lutea, Slight The following examined tissues had no gross observations and no findings microscopically:

-363Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 258 Sex: Female Dose Group: 2 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 05 - Sep - 12 Day o f Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 301.2 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Stomach Stomach Cyst(s), Limiting ridge, Pale, Two, up to 2mm. Squamous Cyst - Limiting Ridge, Focal, Moderate The following examined tissues had no gross observations and no findings microscopically: Ovaries364Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 259 Sex: Female Dose Group: 2 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 06 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 308.4 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Skin Hair loss, ventral cervical to abdominal region and forelimbs. The following examined tissues had no gr oss observations and no findings microscopically: Ovaries365Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluorop ropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 260 Sex: Female Dose Group: 2 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 06 - Sep - 12 Day of Death: 1 0 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 318.9 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Ovaries366Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 261 Sex: Fema le Dose Group: 3 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 309.5 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Spleen Spleen Capsule thickened Capsular/Subcapsular Inflammation, Minimal Adhesions, capsule to adipose tissue. Capsular Thickening , Slight Adhesions/Inflammation/Fibrosis, Minimal Teeth Incisor(s) pale, Lower, Patchy, slight. The following examined tissues had no gross observations and no findings microscopically: Ovaries367Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 262 Sex: Female Dose Group: 3 Phase: Lactatio n Sacrifice Status: Killed for welfare reasons Date of Death: 30 - Aug - 12 Day of Death: 1 Week of Death: 1 Subgroup: 1 Terminal Bodyweight: 304.0 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** Fac con to death Limited Tissues Examined, Pres ent The following examined tissues had no gross observations and no findings microscopically: Ovaries368Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 263 Sex: Female Dose Group: 3 Phase: Gestation Sacrifice Status: Final phase sacrifice Date of Death: 01 - Sep - 12 Day of Death: 25 Week of Death: 4 Subgroup: 1 Terminal Bodyweight: 295.4 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Ovaries369Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 264 Sex: Female Dose Group: 3 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 07 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 303.4 g

PATHOLOGY OBSERVATIONS MACRO PATHOLOGY HIST OPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Ovaries370Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Contr ol 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 265 Sex: Female Dose Group: 3 Phase: Lactatio n Sacrifice Status: Total litter loss Date of Death: 28 - Aug - 12 Day of Death: 1 Week of Death: 1 Subgroup: 1 Terminal Bodyweight: 295.1 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANI MAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Ovaries371Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 266 Sex: Female Dose Group: 3 Phase: Gestation Sacrifice Status: Total litter loss Date of Death: 04 - Sep - 12 Day of Death: 25 Week of Death: 4 Subgroup: 1 Terminal Bodyweight: 282.0 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically: Ovaries372Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropr opene Exposure level (ppm) : 0 198 505 2900 Animal Number: 267 Sex: Female Dose Group: 3 Phase: Lactatio n Sacrifice Status: Total litter loss Date of Death: 30 - Aug - 12 Day of Death: 1 Week of Death: 1 Subgroup: 1 Terminal Bodyweight: 298.1 g

PATHOLO GY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Spleen Spleen Capsule thickened, Anterior pole, two areas, up to 3mm. Capsular Thickening, Slight Adhesions/Inflammation/Fibrosis, Minimal Extramedullary Haemopoiesis, Minimal The following examined tissues had no gross observations and no findings microscopically: Ovaries373Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 268 Sex: Female Dose Group: 3 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 08 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 288.7 g

PATHOLOGY OBSERVATIONS MACRO PATHOLOGY HIST OPATHOLOGY Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findin gs microscopically: Ovaries374Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 269 Sex: Female Dose Group: 3 Phase: Lactatio n Sacrifice Status: Final phase sacrifice Date of Death: 08 - Sep - 12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 277.9 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY ***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED*** The following examined tissues had no gross observations and no findings microscopically: Ovaries375Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 270 Sex: Female Dose Group: 3 Phase: Lactatio n Sacrifice Status: Total litter loss Date of Death: 29 - Aug - 12 Day of Death: 2 Week of Death: 1 Subgroup: 1 Terminal Bodyweight: 304.1 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Ovaries Ovaries Cyst(s), Left, Dark fluid, One, 8mm. Cyst(s), Present The following examined tissues had no gross observations and no findings microscopically:

-376Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Expo sure level (ppm) : 0 198 505 2900 Animal Number: 271 Sex: Female Dose Group: 4 Phase: Gestation Sacrifice Status: Final phase sacrifice Date of Death: 02 - Sep - 12 Day of Death: 25 Week of Death: 4 Subgroup: 1 Terminal Bodyweight: 271.1 g

PATHOLOGY OBS ERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Kidneys Kidneys Irregular surface, Left Cortical Tubular Basophilia, Unilateral, Moderate Pale area(s), Left, A few, subcapsular, diffuse. Cortical Tubular Dilatation, Unilateral, Moderate Spleen Spleen Adhesions, to abdominal wall and surrounding adipose tissue. Capsular Thickening, Focal, Minimal Adhesions/Inflammation/Fibrosis, Minimal Teeth Incisor(s) pale The following examined tissues had no gross observations and no findings microscopically: Ovaries377Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Ma cropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 272 Sex: Female Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 10 - Sep - 12 Day of Death: 54 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 252.2 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Kidneys Kidneys Pelvic dilatation, Right Cortical Tubular Dilatation, Unilateral, Minimal Pelvic Dilatation, Unilateral , Present Ovaries Reduced Size of Corpora Lutea, Slight Spleen Spleen Capsule thickened, Multiple, areas. Capsular/Subcapsular Inflammation, Minimal Adhesions, Anterior pole, capsule to adipose tissue. Capsular Thickening, Slight Adhesions/Inflammation/Fibrosis, Slight Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically:

-378Huntingdon Life SciencesWAG0015

APPENDIX 27 - con tinued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 273 Sex: Female Dose Group: 4 Phase: Lactatio n Sacrifice Status: Total litter loss Date of Death: 29 - Aug - 12 Day of Death: 2 Week of Death: 1 Subgroup: 1 Terminal Bodyweight: 260.1 g

PATHOLOGY OBSERVATIONS MACROPATHOL OGY HISTOPATHO LOGY Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings micro scopically: Ovaries379Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropen e 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 274 Sex: Female Dose Group: 4 Phase: Gestation Sacrifice Status: Final phase sacrifice Date of Death: 29 - Aug - 12 Day of Death: 25 Week of Death: 4 Subgroup: 1 Terminal Bodyweight: 286.8 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Spleen Spleen Enlarged Capsular Thickening, Minimal Extramedullary Haemopoiesis, Moderate Teeth Incisor(s) pale, Lower Vagina Abnormal contents, dark material. The following examined tissues had no gross observations and no findings microscopically: Ovaries380Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

tr ifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 275 Sex: Female Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 10 - Sep - 12 Day of Death: 54 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 24 1.6 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Liver Liver Pale No significant lesions Ovaries Reduced Size of Corpora Lutea, Slight Spleen Spleen Capsule thickened, Multiple, areas, up to 1mm. Capsular/Subcapsular Inflammation, Focal, Minimal Capsular Thickening, Focal, Minimal Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically:

-381Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 276 Sex: Female Dose Group: 4 Phase: Gestation Sacrifice Status: Killed for welfar e reasons Date of Death: 02 - Sep - 12 Day of Death: 24 Week of Death: 4 Subgroup: 1 Terminal Bodyweight: 271.1 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Fac con to death Limited Tissues Examined, Present Kidneys Kidneys Pelvic dilatation, Left Cortical Tubular Basophilia, Unilateral, Slight Pale area(s), Left, Punctate, Multiple, subcapsular . Cortical Tubular Dilatation, Unilateral, Slight Pelvic Dilatation, Unilateral, Present Cortical Tubular Necrosis, Focal, Minimal Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically: Ovaries382Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoroprop ene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 277 Sex: Female Dose Group: 4 Phase: Treatment Sacrifice Status: Final phase sacrifice Date of Death: 10 - Sep - 12 Day of Death: 54 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 250.4 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Ovaries Reduced Siz e of Corpora Lutea, Slight Spleen Spleen Capsule thickened Capsular/Subcapsular Inflammation, Slight Adhesions, capsule to adipose tissue. Capsular Thickening, Moderate Adhesions/Inflammation/Fibrosis, Sli ght Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically:

-383Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 278 Sex: Female Dose Group: 4 Phase: Ge station Sacrifice Status: Final phase sacrifice Date of Death: 30 - Aug - 12 Day of Death: 25 Week of Death: 4 Subgroup: 1 Terminal Bodyweight: 280.8 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Spleen Spleen Adhesions, to omental adipose tissue. Capsular/Subcapsular Inflammation, Minimal Capsular Thickening, Minimal Adhesions/Inflammation/Fibrosis , Slight Extramedullary Haemopoiesis, Slight Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically: Ovaries384Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 279 Sex: Female Dose Group: 4 Phase: Gestation Sacrifice Status: Killed for welfare reasons Date of Death: 06 - Sep - 12 Day of Death: 24 Week of Death: 4 Subgroup: 1 Terminal Bodyweight: 286.2 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Fac con to death Limited Tissues Examined, Present Spleen Spleen Adhesions, to surrounding adipose tissue a nd pancreas. Capsular Thickening, Minimal Adhesions/Inflammation/Fibrosis, Minimal Extramedullary Haemopoiesis, Slight Teeth Incisor(s) pale, Lower The following examined tissues had no gross observation s and no findings microscopically: Ovaries385Huntingdon Life SciencesWAG0015

APPENDIX 27 - continued Request ID: 289382 Macropathology and histopathology - individual findings - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Animal Number: 280 Sex: Female Dose Group: 4 Phase: Gestation Sacrifice Status: Final phase sacrifice Date of Death: 30 - Aug - 12 Day of Death: 25 Week of Death : 4 Subgroup: 1 Terminal Bodyweight: 259.9 g

PATHOLOGY OBSERVATIONS MACROPATHOLOGY HISTOPATHOLOGY Ovaries Reduced Size of Corpora Lutea, Slight Teeth Incisor(s) pale, Lower The following examined tissues had no gross observations and no findings microscopically:

-386Huntingdon Life SciencesWAG0015

APPENDIX 28 Primordial ovarian follicle counts - individual values - F0 generati on Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group 1 1 1 1 1 1 1 1 Animal 241 242 243 244 245 246 247 248 Ovary 1 2 1 2 1 2 1 2 1 2 1 2 1 2 1 2 Section No. 1 3 14 26 23 16 19 4 2 8 34 16 27 28 14 14 50 Mean 8.5 2 4. 5 1 7.5 3 .0 21 .0 2 1.5 21 .0 32 .0 SD 7. 7 8 2.1 2 2.1 2 1.4 1 18. 38 7. 78 9.9 0 25. 46 N 2 2 2 2 2 2 2 2387Huntingdon Life SciencesWAG0015

APPENDIX 28 - continued Primordial ovarian follicle counts - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group 1 1 Animal 249 250 Ovary 1 2 1 2 Section No. 1 19 3 10 6 Mean 11 .0 8 .0 SD 11.3 1 2.8 3 N 2 2388Huntingdon Life SciencesWAG0015

APPENDIX 28 - continued Primordial ovarian follicle counts - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group 2 2 2 2 2 2 2 2 Animal 251 252 253 254 255 256 257 258 Ovary 1 2 1 2 1 2 1 2 1 2 1 2 1 2 1 2 Section No. 1 7 11 15 18 27 32 16 41 3 26 22 31 20 13 18 18 Mean 9.0 16.5 29.5 28.5 14.5 26.5 1 6.5 18.0 SD 2.83 2.12 3.54 17.68 16.26 6.36 4.9 5 0.00 N 2 2 2 2 2 2 2 2389Huntingdon Life SciencesWAG0015

APPENDIX 28 - continued Primordial ovarian follicle counts - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group 2 2 Animal 259 260 Ovary 1 2 1 2 Section No. 1 12 14 26 23 Mean 13.0 24.5 SD 1.41 2.12 N 2 2390Huntingdon Life SciencesWAG0015

APPENDIX 28 - continued Primordial ovarian follicle counts - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group 3 3 3 3 3 3 3 3 Animal 261 262 263 264 265 266 267 268 Ovary 1 2 1 2 1 2 1 2 1 2 1 2 1 2 1 2 Section No. 1 8 10 7 24 26 12 32 19 8 8 9 5 2 22 13 19 Mean 9.0 1 5.5 19 .0 25.5 8 .0 7 .0 12 .0 16.0 SD 1.41 12.0 2 9.9 0 9.19 0.0 0 2.8 3 14.1 4 4.24 N 2 2 2 2 2 2 2 2391Huntingdon Life SciencesWAG0015

APPENDIX 28 - continued Primordial ovarian follicle counts - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group 3 3 Animal 269 270 Ovary 1 2 1 2 Section No. 1 15 3 19 10 Mean 9.0 1 4.5 SD 8.49 6. 36 N 2 2392Huntingdon Life SciencesWAG0015

APPENDIX 28 - continued Primordial ovarian follicle counts - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group 4 4 4 4 4 4 4 4 Animal 271 272 273 274 275 276 277 278 Ovary 1 2 1 2 1 2 1 2 1 2 1 2 1 2 1 2 Section No. 1 6 13 9 5 20 7 11 5 40 11 20 18 15 18 3 13 Mean 9.5 7 .0 1 3.5 8 .0 2 5.5 19 .0 1 6.5 8 .0 SD 4.9 5 2.8 3 9. 19 4.2 4 20.5 1 1.4 1 2.1 2 7. 07 N 2 2 2 2 2 2 2 2393Huntingdon Life SciencesWAG0015

APPENDIX 28 - continued Primordial ovarian follicle counts - individual values - F0 generation Group : 1 2 3 4 Compound : Control 2 - br omo - 3,3,3 - trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene 2 - br omo - 3,3,3 -

trifluoropropene Exposure level (ppm) : 0 198 505 2900 Group 4 4 Animal 279 280 Ovary 1 2 1 2 Section No. 1 19 28 28 18 Mean 2 3.5 23 .0 SD 6. 36 7. 07 N 2 2Huntingdon Life Sciences WAG0015

394 

### Annex 1 Certificate of analysisHuntingdon Life Sciences WAG0015

395Huntingdon Life Sciences WAG0015

396 

### Annex 2 Aerosol technology and inhalation analysis reportHuntingdon Life Sciences WAG0015

397 

### Working for a better future



## Aerosol Technology R eport

HLS s tudy number: WAG00 15 Author A. Grasiewicz Version ID: Final Issue date: 06 March 2013Huntingdon Life Sciences WAG0015

398 

# Table of Contents Signature Page ................................................................................................................ ... 399 1. Pre-study Atmosphere Characterization ..................................................................... 400 2. Test Article Handling and Processing for Inhalation Generation ................................. 400 3. Exposure System ........................................................................................................ 400 4. Administration ............................................................................................................. 40 1 5. Atmosphere Analysis .................................................................................................. 401 5.1 Vapour Concentration ........................................................................................... 401 5.2 Flow diagram of the analytical method for the analysis of 2-bromo-3,3,3- trifluoropropene in test atmosphere samples .......................................................... 401 5.3 Nominal Concentration ......................................................................................... 402 5.4 Chamber Temperature ......................................................................................... 402 5.5 Chamber Humidity ................................................................................................ 402 6. Data and Calculations ................................................................................................. 402 7. Results and Discussion ............................................................................................... 402 7.1 Aerosol Concentrations ........................................................................................ 402 7.2 Chamber Temperature ......................................................................................... 403 7.3 Chamber Humidity ................................................................................................ 403 

# List of Tables Table 1 Chamber atmosphere concentration of 2-bromo-3,3,3-trifluoropropene – daily means and individual exposure values – Group 2....................................... 404 Table 2 Chamber atmosphere concentration of 2-bromo-3,3,3-trifluoropropene – daily means and individual exposure values – Group 3....................................... 406 Table 3 Chamber atmosphere concentration of 2-bromo-3,3,3-trifluoropropene - daily means and individual exposure values – Group 4...................................... . 408 Table 4 Chamber Temperatures - daily mean values .............................................. 410 Table 5 Chamber Humidity - daily mean values ....................................................... 412 

# List of Figures Figure 1 Schematic of Inhalation Exposure System .................................................. 414Huntingdon Life Sciences WAG0015

399 

# Signature Page Aerosol Technology ReportHuntingdon Life Sciences WAG0015

400 

# 1. Pre -study

A tmosphere Characterization Inhalat io n exposure system characterized including determinat io ns of: Achieved vapour concentration Spatial d istribut io n around the chamber 

# 2. Te st Article Handling and Processing for Inhalation Generation As supplied, 2 - Bromo- 3,3,3 - trifluoro -1- propene was easily vapo uri sed using warmed air th rough a sintered glass vapo uri ser. 

# 3. Exposure Sy

stem A schemat ic o f the exposure system is pres ented in Figure 1. Com ponents of the exposure system : Di

rected flow Whole body

exposure chamber: 0.75 m

3 

### chamber of stainless steel and glass construction Animal Restraint: Wire mesh modular caging Aerosol Generator: Sint ered glass vaporiser for Groups 2 to 4 all in a water bath set at 40°C Sy ringe driver, polypropylene syringe for Group 2 Liquid reservoi

r and feed rate used to alter concentration Groups 3 and 4 Test groups - 30 L/min Diluent Airflow: From in- house co mpressed air system

– breathing qualit y Group 1 - 149 L/min Group 2 to 4 - 119 L/min Extract Ai

rflow: Drawn by a fan extract system Drawn fro m base of exposure system Controlled by a gate clamp situated at the rear of the chamber Airflow Monitoring: In- line flowmeters monitored continuously for generation and diluent ai rflows Chamber pressure different ial mo nit ored by Magnahelic set at - 1 to

- 5 mm H 2 

### O to ensure a slight negat ive pressure within the chamber ~ 150 L/min.Huntingdon Life Sciences WAG0015

401 

# 4. Ad ministration Animals exposed for 6 hours per day, 7 days a week Exposure system

operational

parameters moni

tore d conti

nuously Airflows documented init ially and moni

tored during exposure 

# 5. At mosphere Analysis 5.1 Vapour Concentration Vapour sam

ples co llected as fo llo ws: Sam ple f requency – minimum o f 3 sam ples/group/exposure Sam ple location – representative animal level Sam ple posi tion remained constant Sam ple analysis Infra - red spectrophotometer 

## 5.2 Flow diagram of the analytical method for the analysis of 2- bromo - 3,3,3 -trifluoropropene in test atmosphere samples Sample Assay Test atmosphere sample (Gas bag or chamber)

 Sample directly using the IR spectrophotometer

 Perform a single measurement on the IR

spectrophotometer Calibration Prepare a set of calibration standards by direct addition of a suitable volume of test item directly into gas bags containing a suitable volume of air

 Prepare a set of calibration standards for analysis at nominal concentrations of 146, 388, 1167, 2042, 2918

and 3880 ppm  Measure the response of each calibration standard using the IR spectrophotometer forcalibration IR conditions Instrument: MIRAN SapphIRe model 205B Path length: Short (0.5m) Wavelength: 10.699 μm The IR spectrophotometer was calibrated using external standards. Absorbance measurements acquired by the IR spectrophotom eter were used to calculate analysed concentration (ppm) using a 2 nd order fit subjected to least squares regression analysis. Standards were shared with another concurrent and related study.Huntingdon Life Sciences WAG0015

402 

## 5.3 Nominal Concentration A mean daily no minal concentration was calculated for all test groups from the mass of test article used over the exposure period and the airflo w through the exposure chamber. The ideal gas equat io n was used wit h the mo lecular weight of the test article and the measured chamber condit io ns to c om pute the vo lume of vapour produced from the mass of test article used. This data is not reported but retained in the raw data. 

## 5.4 Chamber Temperature Moni tored using a liquid in glass thermometer . 

## 5.5 Chamber Humidity Moni tored using wet and dry

liquid in glass thermometers, wi th h umidit y calculated using proprietary

tables . 

# 6. Data and Calculations To minimize the cumulat ive errors from repeated rounding of numbers, data in this report has been calculated continuously using unrounded numbers and only rounded fo r print ing. Consequent ly, presented data may include rounding errors in the last significant figure. All data not presented in this report is retained in the study data files and archives. 

# 7. Results and Discussion 7.1 Aerosol Concentrations Individual exposure concentrations and daily means are presented in Table s 1 to 3 and summarized below: Group C oncentration ( ppm )

Target Mean 2 200 19 8 3 500 505 4 3 000 2 900 

### All groups were close to target . Addit io nal samples were taken for confirmat io n of atypical resul ts and after minor generat io n rate adjust me nt s. All samples co llected for Group 1 were negative for the presence of test article . T his informat io n is not tabulated but is retained in the study data files .Huntingdon Life Sciences WAG0015

403 

## 7.2 Chamber Temperature The chamber temperature was moni tored continuously for all groups and recorded at 30 minute intervals. D aily means are presented in Table 4 and were within expected parameters for this ty

pe of exposure system. 

## 7.3 Chamber Humidity The chamber humidit y was derived from wet and dry

bulb thermometers f

or all groups. These tem

peratures were recorded at 30 minute intervals and the subsequent humidit y determined fro m propriatory tables. D aily

means are presented in Table 5 and were within expected parameters for this ty pe of exposure system.Huntingdon Life Sciences WAG0015

404 

### Table 1 Chamber atmosphere concentration of

2-bromo-3,3,3 -trifluoropropene – daily means and individual exposure values – Group 2 Exposure Chamber concentration (ppm) Day Sample

1 Sample

2 Sample

3 Sample

4 Sample

5 Sample

6 Daily mean 1 63 92 115 147 147 178 124 2 166 92 187 174 155 3 153 138 126 71 246 178 152 4 263 206 210 226 5 201 206 212 206 6 115 203 212 177 7 189 206 208 201 8 86 176 195 187 161 9 199 180 203 194 10 203 203 206 204 11 191 208 210 203 12 201 206 206 204 13 183 192 194 190 14 188 190 190 189 15 P 181 194 196 190 16 G 186 192 196 191 17 183 186 196 188 18 181 186 192 186 19 181 192 194 189 20 190 192 192 191 21 192 192 192 192 22 190 195 197 194 23 188 195 195 193 24 188 190 197 192 25 186 1 188 1 190 1 188 1 26 190 190 197 192 27 192 194 180 189 28 202 209 207 206 29 213 215 213 214 30 205 211 217 211 31 192 211 204 202 32 180 1 180 1 182 1 181 1 33 202 215 219 212 34 211 217 219 216 35 215 215 221 217 36 191 197 209 199 37 189 201 203 198 38 L 194 190 188 191 39 190 192 192 191 40 200 204 206 203 41 208 210 214 211 42 202 204 212 206 43 202 206 204 204 44 204 210 212 209 45 202 206 208 205 46 204 208 218 210 47 206 208 204 206 48 205 220 216 214Huntingdon Life Sciences WAG0015

405 

### Table 1 Chamber atmosphere concentration of

2-bromo-3,3,3 -trifluoropropene – daily means and individual exposure values – Group 2 (continued) Exposure Chamber concentration (ppm) Day Sample

1 Sample

2 Sample

3 Sample

4 Sample

5 Sample

6 Daily mean 49 207 212 216 212 50 203 210 218 210 51 216 216 220 217 52 216 220 220 219 53 223 229 203 218 5 4 205 220 225 217 55 203 207 210 207 56 175 193 201 190 Mean 19 8 sd 18 . 2 CV (%) 9.2 1 For exposures 4.4 and 5.4 the sample s have been excluded due to standard check bag failures. The samples and test article usage were within expected tolerances. sd Standard deviation CV (%) Coefficient of variation (sd × 100/mean) P Pairing commenced G First possible commencement of gestation phase L First possible commencement of lactation phaseHuntingdon Life Sciences WAG0015

406 

### Table 2 Chamber atmosphere concentration of

2-bromo-3,3,3 -trifluoropropene – daily means and individual exposure values – Group 3 Exposure Chamber concentration (ppm) Day Sample

1 Sample

2 Sample

3 Sample

4 Sample

5 Sample

6 Samp le

7 Daily mean 1 308 299 786 571 499 471 839 539 2 417 551 456 475 3 242 425 523 397 4 471 619 518 536 5 346 299 735 460 6 394 456 469 440 7 490 566 516 524 8 458 486 490 478 9 503 392 564 486 10 559 551 499 536 11 546 469 518 511 12 407 687 531 467 523 13 462 522 518 501 14 495 409 608 362 469 15 P 329 453 453 412 16 G 475 469 608 517 17 493 437 545 492 18 471 424 587 494 19 460 489 531 493 20 549 505 539 531 21 489 354 503 539 471 22 483 451 489 474 23 517 567 580 555 24 549 555 551 552 25 510 1 480 1 505 1 498 1 26 430 517 535 537 505 27 453 453 578 495 28 476 566 615 552 29 498 459 541 499 30 512 516 549 526 31 413 465 472 539 472 32 611 1 533 1 508 1 482 1 534 1 33 613 615 642 599 586 541 506 586 34 111 548 409 475 437 810 756 507 35 347 488 453 429 36 465 455 518 479 37 536 494 500 510 38 L 458 461 535 485 39 513 605 607 542 537 561 40 517 495 482 498 41 517 493 503 504 42 511 466 501 493 43 451 507 619 557 534 44 491 852 575 509 607 45 540 476 476 497 46 478 474 511 488 47 501 466 523 497 48 540 500 487 509Huntingdon Life Sciences WAG0015

407 

### Table 2 Chamber atm osphere concentration of

2-bromo-3,3,3 -trifluoropropene – daily means and individual exposure values (continued) – Group 3 Exposure Chamber concentration (ppm) Day Sample

1 Sample

2 Sample

3 Sample

4 Sample

5 Sample

6 Sample

7 Daily mean 49 7 43 511 481 571 577 50 716 593 558 520 597 51 476 476 450 467 Mean 505 sd 43 . 1 CV (%) 8. 5 1 For exposures 4.4 and 5.4 the samples have been excluded due to standard check bag failures. The samples and test article usage were within expected tolerances. sd Standard deviation CV (%) Coefficient of variation (sd × 100/mean) P Pairing commenced G First possible commencement of gestation phase L First possible commencement of la ctation phaseHuntingdon Life Sciences WAG0015

408 

### Table 3 Chamber atmosphere concentration of

2-bromo-3,3,3 -trifluoropropene - daily means and individual exposure values – Group 4 Exposure Chamber concentration (ppm) Day Sample

1 Sample

2 Sample

3 Sample

4 Sample

5 Sample

6 Daily mean 1 3175 3332 3191 3092 2817 3121 2 2860 2584 2362 2602 3 2000 2303 2444 2249 4 4184 3939 2303 2110 3134 5 1990 2293 3007 2430 6 3095 2975 3356 3142 7 2697 2944 2707 2783 8 2046 2224 2771 2278 2330 9 2586 386 1 3148 3228 2987 10 3372 3413 2238 3008 11 3645 3252 2631 3276 3201 12 2871 3856 3268 1575 2864 2887 13 3274 3030 3111 3138 14 3030 3415 3518 3184 3287 15 P 2426 3316 3297 3013 16 G 3297 3027 3349 3224 17 3067 2806 2788 2887 18 2469 2955 2437 2620 19 2845 3200 3168 3071 20 3043 2858 3095 2999 21 3422 3016 1707 2994 2888 2805 22 2301 3087 2745 2711 23 2977 3696 2734 3136 24 2828 3303 3068 3066 25 2983 2 2193 2 2729 2 3071 2 2744 26 2615 2745 304 0 2874 2819 27 2711 2795 3377 2961 28 2766 3262 2401 3341 2943 29 2908 2751 3256 2972 30 3224 3243 3181 3216 31 2876 2737 2795 2803 32 2213 2 2861 2 2809 2 3150 2 2758 33 2483 3181 2714 2546 3458 2876 34 719 2939 2550 3526 3387 2958 2680 35 2345 2435 3136 2917 2708 36 2476 2936 2721 2606 2684 2685 37 2338 2710 2826 2625 38 L 2126 3448 3376 3025 2998 2995 39 2764 2912 3112 2929 40 2623 2885 2610 2706 41 2620 2699 2919 2746 42 2717 3088 2880 2895 43 2638 2707 3287 2877 44 3214 3106 2955 3092 45 3472 2924 2727 2709 2958 46 2722 2725 2725 2724 47 4103 3168 2717 2702 3173 48 3217 2953 2783 2984Huntingdon Life Sciences WAG0015

409 

### Table 3 Chamber atmosphere concentration of

2-bromo-3,3,3 -trifluoropropene - daily means and individual exposure values (continued) – Group 4 Exposure Chamber concentration (ppm) Day Sample

1 Sample

2 Sample

3 Sample

4 Sample

5 Sample

6 Daily mean 49 3209 2796 2791 2932 50 2877 3179 2806 2954 51 3017 2882 2838 2912 52 2793 2801 2827 2807 53 2958 3136 2927 3007 Mean 2899 sd 223 CV (%) 7. 7 1 Value omitted from daily mean due to sample taken when pressure ve ssel was empty, vessel refilled and sample repeated . 2 For exposures 4.4 and 5.4 the samples have been excluded due to standard check bag failures. The samples and test article usage were within expected tolerances. sd Standard deviation CV (%) Coefficient of variation (sd × 100/mean) P Pairing commenced G First possible commencement of gestation phase L First possible commencement of lactation phaseHuntingdon Life Sciences WAG0015

410 

### Table 4 Chamber Temperatures - daily mean values Exposure Day Chamber Temperature (°C )

Group 1 Group 2 Group 3 Group 4 1 25.9 23 .9 23.0 23.8 2 24.8 23.9 23.5 23.6 3 22.7 22.5 21.5 20.8 4 22.8 22.0 22.0 21.0 5 26.0 23.7 23.0 24.3 6 24.7 23.4 23.2 24.2 7 24.1 22.5 22.7 24.3 8 24.9 22.8 21.9 23.7 9 25.3 22.9 21.8 23.4 10 20.7 20.7 19.5 19.2 11 21.3 20.7 19.6 20.7 12 25.0 23 .8 22.8 24.2 13 25.4 24.0 23.6 24.4 14 24.8 24.3 23.2 24.3 15 25.1 24.5 23.3 24.2 16 24.8 24.2 23.3 25.0 17 22.3 23.5 22.8 23.5 18 23.0 23.2 22.8 23.2 19 25.6 24.8 23.4 24.2 20 25.2 24.3 23.8 24.4 21 25.3 24.6 23.7 24.8 22 25.5 24.8 23.9 25.0 2 3 25.9 24.5 23.6 24.4 24 22.2 22.7 22.2 22.9 25 22.4 22.8 22.3 22.9 26 25.5 24.9 23.9 24.7 27 25.5 24.3 23.8 25.4 28 25.7 23.8 23.4 24.8 29 25.4 23.8 23.0 25.3 30 25.3 24.3 22.9 24.6 31 22.5 20.5 20.9 22.6 32 22.2 21.8 21.7 21.9 33 26.2 23.8 22.9 24.6 34 25.7 22.7 22.4 24.9 35 25.8 22.6 22.8 25.0 36 26.2 23.8 23.4 24.0 37 25.8 23.6 23.2 23.8 38 23.1 21.3 20.4 22.0 39 21.8 21.3 20.6 21.3 40 25.6 24.1 23.4 25.3 41 26.1 24.3 24.3 25.8 42 25.8 23.8 23.8 24.9 43 25.8 23.8 23.7 24.5 44 26.5 2 4.7 23.3 25.0 45 23.9 23.1 22.0 23.0 46 24.1 23.8 22.5 22.9 47 26.6 24.7 26.6 24.3 48 25.9 24.0 25.9 24.9Huntingdon Life Sciences WAG0015

411 

### Table 4 C ha mber Temperatures - daily mean values (continued) Exposure Day Chamber Temperature (°C )

Group 1 Group 2 Group 3 Group 4 49 25.5 23.7 25.5 24.4 50 25.6 24.0 25.6 25.1 51 22.3 24.7 24.2 52 20.2 21.9 53 21.1 22.1 54 25.6 55 24.2 56 25.0 Mean 24.7 23.4 23.0 23.8 sd 1.5 1.3 1.4 1.4 CV (%) 6.1 5.4 6.0 5.9 sd Standard deviation CV (%) Coefficient of variation (sd × 100/mean)Huntingdon Life Sciences WAG0015

412 

### Table 5 Chamber Humidity - daily mean values Exposure Day Chamber Humidity ( % )

Group 1 Group 2 Group 3 Group 4 1 30.9 49.6 32.9 47.2 2 52.0 51.2 29.9 51.2 3 41.2 53.8 53. 8 64.8 4 28.5 42.7 30.6 60.3 5 30.2 37.5 40.2 39.2 6 72.5 46.2 48.6 60.2 7 50.9 48.0 40.2 53.1 8 34.5 50.1 48.9 78.0 9 35.8 36.5 51.2 68.1 10 29.1 41.9 50.5 63.0 11 29.5 43.6 49.8 55.3 12 49.9 31.6 41.6 51.6 13 28.3 37.1 47.8 53.1 14 44.2 29.1 3 8.1 64.4 15 35.8 30.8 47.9 50.9 16 41.5 36.5 44.0 54.9 17 31.1 24.1 34.5 43.9 18 27.7 28.6 33.8 42.9 19 34.5 30.7 44.6 57.0 20 39.7 35.2 47.7 49.9 21 36.7 38.5 44.8 49.8 22 34.8 38.0 49.4 48.2 23 39.0 35.0 44.4 57.7 24 39.8 24.4 37.9 44.4 25 35 .8 25.2 33.8 44.3 26 33.5 32.0 41.6 50.2 27 33.0 39.0 42.9 50.7 28 31.5 36.8 49.2 53.8 29 34.0 39.6 38.3 41.8 30 40.8 38.3 42.4 47.8 31 34.6 35.8 60.2 35.5 32 31.9 37.8 33.2 55.6 33 35.2 43.5 41.2 56.5 34 29.8 51.4 60.1 44.7 35 29.5 51.6 53.8 45. 5 36 28.7 41.3 46.3 63.8 37 32.1 43.8 50.0 61.0 38 26.9 64.0 64.0 26.9 39 31.0 57.4 57.4 31.0 40 29.2 32.2 43.6 47.8 41 25.9 29.0 38.5 48.4 42 29.8 37.0 38.1 49.3 43 27.3 32.6 41.1 55.5 44 31.3 30.2 40.6 48.4 45 28.3 32.7 32.7 47.8 46 27.3 23.3 37.0 55.2 47 33.0 33.0 44.1 54.7 48 29.0 38.4 44.4 47.2Huntingdon Life Sciences WAG0015

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### Table 5 C hamber Humidity - daily mean values (continued) Exposure Day Chamber Humidity ( % )

Group 1 Group 2 Group 3 Group 4 49 32.1 32.5 50.1 58.9 50 32.6 29.2 40.3 44.5 51 27.8 39.6 42.6 52 45.2 44.4 53 29.5 43.0 54 26.8 55 49.5 56 32.7 Mean 34.6 37.9 43.9 51.1 sd 8.2 9.0 7.8 9.1 CV (%) 23.6 23.8 17.7 17.8 sd Standard deviation CV (%) Coefficient of variation (sd × 100/mean)Huntingdon Life Sciences WAG0015

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### Figure 1 Schematic of Inhalation Exposure System

D A B

F C E

F

H I

J K Front view Side view

L Generation Trolley

G M N P Q O



### Key A Who le body

chamber J Water bath B Test material delivery tubing K Generat io n flo wmeters C El utriator L Control valves D Secondary

generation trolley M Balance for more accurate weighing E Magnahelics N Harvard Sy ringe driver F Pressure Vessels O Sam pling port G Load cells P Extract plenum inside chamber H Air - line co il Q Drain for washing down I Sint ered glass vaporiser Closed during exposureHuntingdon Life Sciences WAG0015

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### Annex 3 Pathology reportHuntingdon Life Sciences WAG0015

Page 1 of 9 

### Working for a better futur e



## PATHOLOGY REPORT 2-bromo-3,3,3-trifluoropropen e: Reproductive/Developmental Toxicity Screening Study in the CD Rat by Inhalation Administration

HLS study number: WAG0015 Version ID: Final Issue date: 13 November 2012Huntingdon Life Sciences WAG0015

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# Table of Contents Signature Page ................................................................................................................ ....... 2 1. Introduction .................................................................................................................. .. 4 1.1 Objective ............................................................................................................ 4 2. Results ....................................................................................................................... .... 4 2.1 Decedents .......................................................................................................... 4 2.2 Macroscopic Pathology ...................................................................................... 5 2.2.1 Animals killed after scheduled treatment period ..................................... 5 2.3 Microscopic Pathology ....................................................................................... 6 2.3.1 Animals killed after scheduled treatment period ..................................... 6 3. Discussion .................................................................................................................... .. 7 4. Conclusion .................................................................................................................... . 8 5. Appendix I .................................................................................................................... .. 9Huntingdon Life Sciences WAG0015

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# 1. Introduction 1.1 Objective Initial screening assessment of the influence of 2-bromo-3,3,3-trifluoropropene when administered by inhalation administration on reproductive performance and development in the CD rat. Group Treatment Achieved concentration (ppm) Number of animals Main study Male Female 1 Control 0 10 10 2 2-bromo-3,3,3- trifluoropropene 198 10 10 3 2-bromo-3,3,3- trifluoropropene 505 10 10 4 2-bromo-3,3,3- trifluoropropene 2900 10 10 

# 2. Results 2.1 Decedents Two females exposed to 2900 ppm, animals 276 and 279, were killed for welfare reasons on Day 24 of the gestation phase and one femaleexposed to 505 ppm , animal 262, was killed for welfare reasons on Day 1 of the lactation phase. No histopathological findings were seen in the ovaries of these three animals. Animal 276 had multiple but unilateral histopathological changes in the kidneyand animal 279 had capsular thickening, adhesions/inflammation/fibrosis of the spleen; in both cases the findings observed could not be considered as a factor contributing to death.Huntingdon Life Sciences WAG0015

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## 2.2 Macroscopic Pathology 2.2.1 Animals killed after scheduled treatment period The macroscopic examinations performed in the F0 animals revealed intergroup differences in the prostate, spleen, incisor teeth and skin. Prostate A reduction in size of the prostate was seen inall males exposed to 2900 ppm, in the majority of males exposed to 505 ppm and in one male exposed to 198 ppm. Summary of findings in the prostate of F0 animals after scheduled treatment period

Group/sex 1M 2M 3M 4M Exposure Level (ppm) 0 198 505 2900 Small 0 1 7 10 Number of animals examined 10 10 10 10 

### Spleen Capsular thickening was seen inthe spleen of the majority of males exposed to 2900, 505 or 198 ppm and in occasional females exposed to 2900 or 505 ppm. Capsular adhesions were seen in occasional treated males across all groups, in a few females exposed to 2900 ppm and one female exposed to 505 ppm.

Summary of findings in the spleen of F0 animals after scheduled treatment period Group/sex 1M 2M 3M 4M 1F 2F 3F 4F Exposure Level (ppm) 0 198 505 2900 0 198 505 2900 Capsule thickened 0 8 6 9 0 0 2 3 Adhesions 0 2 3 4 0 0 1 4 Number of animals examined 10 10 10 10 10 10 9 8 

### Teeth Pale incisor teeth were seen in all male and female animals exposed to 2900 ppm, in the majority of males and three females exposed to 505 ppm and in one control male.

Summary of findings in the teeth of F0 animals after sche duled treatment period Group/sex 1M 2M 3M 4M 1F 2F 3F 4F Exposure Level (ppm) 0 198 505 2900 0 198 505 2900 Incisor(s) pale 1 0 6 10 0 0 3 8 Number of animals examined 10 10 10 10 10 10 9 8Huntingdon Life Sciences WAG0015

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### Skin An increased incidence of hair loss was noted in occasional males exposed to 2900ppm.

Summary of findings in the skin of F0 animals after scheduled treatment period Group/sex 1M 2M 3M 4M 1F 2F 3F 4F Exposure Level (ppm) 0 198 505 2900 0 198 505 2900 Hair loss 0 1 2 4 2 2 0 0 Number of animals examined 10 10 10 10 10 10 9 8 

### The nature and incidence of all other findings were consis tent with the common background of macroscopic changes seenat these laboratories. 

## 2.3 Microscopic Pathology 2.3.1 Animals killed after scheduled treatment period Treatment related findings Changes related to treatment with 2-bromo- 3,3,3-trifluoropropene were seen in the F0 ovaries. Ovary A slight reduction in the size of corpora lutea wa s seen in the ovaries of 4 females exposed to 2900 ppm and in a single female exposed to 198 ppm. Summary of findings in the ovary of F0 animals after scheduled treatment period

Group/sex 1F 2F 3F 4F Exposure Level (ppm) 0 198 505 2900 Reduced Size of Corpora Lutea Slight 0 1 0 4 Total 0 1 0 4 Number of tissues examined 10 10 9 8 

### Findings of an uncertain relationship to treatment Findings of an uncertain relationship to treatmentwere identified in the spleen. This tissue was examined microscopically only when macroscopic abnormalities had been observed at necropsy. No spleen from control animals was examined. Spleen Minimal to moderate capsular/subcapsular inflammation and/or capsular thickening and/or adhesions/inflammation/fibrosis were observed in the majority of treated males exposed to 198, 505 or 2900 ppm and in occasional fe males exposed to 505 or 2900 ppm.Huntingdon Life Sciences WAG0015

Page 7 Summary of findings of an uncertain relationship totreatment in the spleen of F0 animals after scheduled treatment period Group/sex 1M 2M 3M 4M 1F 2F 3F 4F Exposure Level (ppm) 0 198 505 2900 0 198 505 2900 Capsular/Subcapsular Inflammation Minimal 0 5 3 3 0 0 1 3 Slight 0 3 2 2 0 0 0 1 Moderate 0 0 1 2 0 0 0 0 Total 0 8 6 7 0 0 1 4 Capsular Thickening Minimal 0 4 4 2 0 0 0 4 Slight 0 0 2 4 0 0 2 1 Moderate 0 0 0 1 0 0 0 1 Total 0 4 6 7 0 0 2 6 Adhesions/Inflammation/Fibrosis Minimal 0 3 2 2 0 0 2 1 Slight 0 2 1 3 0 0 0 3 Moderate 0 0 1 1 0 0 0 0 Total 0 5 4 6 0 0 2 4 Number of tissues examined 0 8 7 9 0 0 2 6 

### The incidence and distribution of all other findings were consistent with the common background seen at these laboratories. 

# 3. Discussion All females exposed to 2900 ppm and half of those exposed to 505 ppm were killed at a different time during the study as they failed tomate (sacrificed on Day 25 after the last paring) or failed to litter (sacrificed on Day 25 after mating) or lost the litter post-partum (sacrificed on the Day of litter loss); therefore, none performe d as expected up to the end of the study on Day 10 of lactation. The examination of the ovaries from the F0 generation females reve aled the presence of histopathological changes attrib utable to treatment. A reduction in size of corpora lutea was observed in the ovaries of four females e xposed to 2900 ppm and one female exposed to 198 ppm. The corpora lutea were quite small an d appeared to be not newly formed. As this change was not consistent w ith the increasing exposure leve l across the treated groups, the presence in a single female exposed to 198 ppm was considered to be incidental. There was no apparent difference in the average number of corpora lutea in the treated groups when compared with controls, as reported in Appendix I. This count did not take account of differences in size of corpora lutea, however, as has been reported. No histopathological findings were seen to account fo r the irregular oestrus cycle or the prolonged gestation length as reported in the in life phase.Huntingdon Life Sciences WAG0015

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### The histopathological examination of testes and epididymides from the F0 generation animals performed after scheduled treatment period reve aled no findings attributable to treatment with the test article 2- bromo-3,3,3-trifluoropropene. The seminiferous tubules were evaluated with respect to their stage in the spermatogenic cycle and the integrity of the various cell types present withinthe different stages. No significant cell or stage-specific abnormalities were seen in the sections of testes either stained with haematoxylin & eosin (H&E ) or periodic acid Schiff (PAS). Evaluation of the epididymides did not reveal any changes in leukocyte infiltration, cell populations, or sperm numbers. There were no fi ndings to correlate with the organ weight variation reported at necropsy. An increased incidence of small prostates wa s observed at the macroscopic examination. This was consistent with a reduc tion in prostate weight seen w ith increasing exposure level. No microscopic correlate for the small prosta tes could be identified. A low incidence of inflammatory changes was observed in all groupsincluding controls and was considered to be incidental as these types of changes are occasionally seen in rats of this age. Capsular thickening and adhesions were observed in the spleen of the majority of treated males and occasional treated females at the macroscopic examination. Most of these macroscopic observations correlated with findings of caps ular/subcapsular inflammation and/or capsular thickening and/or adhesions/inflammation/fibros is seen at the microscopic examination and are likely to be related to treatment. However, as no controls and only tissues with macroscopic changes were sample d and microscopically examined, conclusions as to the significance of these changes could not be made. The significance of the increased incidence of paleincisors and hair loss of the skin observed at the macroscopic examination could not be cl arified as these lesions were not examined microscopically. Pale incisors and hair loss of the skin are not routinely presented fo r microscopic evaluation as such changes do not usually correlate with changes seen histopathologically. These tissues are also not routinely require d to be examined for reprodu ctive developmental screening studies. 

# 4. Conclusion The microscopic evaluation on this study reveal ed the presence of histopathological changes related to 2-bromo-3,3,3-trifluoropropene only in the ovaries of animals exposed to 2900 ppm.Huntingdon Life Sciences WAG0015

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# 5. Appendix I Corpora Lutea Counts Group/ Animal No. L Ovary Level 1 R Ovary Level 1 Total CL 1F/241 4 9 13 1F/242 9 11 20 1F/243 7 12 19 1F/244 3 13 16 1F/245 8 11 19 1F/246 12 14 26 1F/247 5 5 10 1F/248 5 13 18 1F/249 13 9 22 1F/250 11 8 19 2F/251 10 16 26 2F/252 15 6 21 2F/253 8 11 19 2F/254 21 13 34 2F/255 5 15 20 2F/256 12 13 25 2F/257 16 18 34 2F/258 18 18 36 2F/259 17 11 28 2F/260 15 7 22 3F/261 12 7 19 3F/262 8 8 16 3F/263 12 13 25 3F/264 9 12 21 3F/265 3 2 5 3F/266 4 9 13 3F/267 4 14 18 3F/268 11 12 23 3F/269 13 6 19 3F/270 12 8 20 4F271 9 11 20 4F272 16 11 27 4F273 6 4 10 4F274 8 5 13 4F275 5 13 18 4F276 4 11 15 4F277 7 10 17 4F278 8 5 13 4F279 5 7 12 4F280 12 16 28 Key: L= Refer to the ovary on the Left of each slide; R= Refer to the ovary on the Right of each slide. CL= Corpora LuteaHuntingdon Life Sciences WAG0015

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### Annex 4 GLP compliance statementsHuntingdon Life Sciences WAG0015

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### HUNTINGDON RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2011Huntingdon Life Sciences WAG0015

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### HUNTINGDON RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2012Huntingdon Life Sciences WAG0015

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### EYE RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2011Huntingdon Life Sciences WAG0015

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### EYE RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2012