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Working for a better future

REPORT

2-bromo-3,3,3-trifluoropropene: Reproductive/Developmental

Toxicity Screening Study in the CD Rat by Inhalation

Administration

HLS study number: WAG0015

Version ID: Final

Issue date: 22 July 2013
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Details of Sponsor and Test Facilities

Sponsor American Pacific Corporation

3883 Howard Hughes Parkway

Suite 700

Las Vegas

Nevada 89169

USA

Test facility Huntingdon Life Sciences

Primary location Huntingdon Research Centre

Woolley Road

Alconbury

Huntingdon

Cambridgeshire

PE28 4HS

UK

Test facility Huntingdon Life Sciences

Oestrous cycles, Eye Research Centre

Sperm analysis Eye

Histology, Suffolk

Histopathology IP23 7PX

UK
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Table of Contents

Details of Sponsor 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 administration................................................................................... 14

1.5 Treatment groups and doses .......................................................................... 14

2. Experimental Procedure............................................................................................. 15

2.1 Study schedule and structure.......................................................................... 15

2.1.1 Duration of treatment........................................................................... 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 Water 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................................................................................................... 28
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2.8.2 Bodyweights........................................................................................ 28

2.8.3 Food consumption............................................................................... 28

2.8.4 Water 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 Water 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 scheduled 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 ......................................................................................................... 49
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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 Bodyweight - group mean values for females during lactation......................53

Figure 5 Bodyweight - group mean values for offspring .............................................54

List of Tables

Table 1 Summary of adult performance....................................................................56

Table 2 Clinical signs - group distribution of observations.........................................57

Table 3 Signs associated with dosing - group distribution 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 females during lactation...........87

Table 11 Water consumption - group mean values for males and before pairing for 

females ........................................................................................................88

Table 12 Water consumption - group mean values for females during gestation ........90

Table 13 Water 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 and 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 ............................................................................................103
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Table 24 Bodyweight and bodyweight 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 Macropathology - 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 - individual 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 - Individual 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 values for females after mating...................248

Appendix 11 Food consumption - individual values for females during lactation .............250

Appendix 12 Water consumption - individual values for males before and after pairing and 

females before mating - F0 generation (mL/animal/day)........................................252

Appendix 13 Water consumption - individual values for females after mating.................256

Appendix 14 Water 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 ................................................................................289
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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 analysis.................................................................................394

Annex 2 Aerosol technology and inhalation analysis report......................................396

Annex 3 Pathology report.........................................................................................415

Annex 4 GLP compliance statements ......................................................................425


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Contributing Scientists

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 CBiol MSB

Study Director

David P Myers BSc (Hons) PhD IDT

Senior Toxicologist

Steven J Crome BSc MSc

Head of Toxicologists (inhalation aspects)

Formulation analysis

Caroline Finucane PhD

Head of Formulation & Inhalation Analysis

Aerosol technology

Simon A Moore BSc (Hons) PhD MRSC

Head of Aerosol Technology

Oestrous cycles and mating performance

Martin J Collier BSc

Behavioural Scientist

Sperm analysis

Clare Bowden BSc (Hons) CBiol MSB

Operational Head, Sperm Analysis & Embryo Culture

Pathology

Miledi Azzalin DVM MRCVS

Pathologist

Department of Pathology

Statistics

Gareth D Thomas BSc (Hons)

Head of Statistics and Data Management
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Summary

The objective of this study was an initial screening assessment of the influence of 

2-bromo-3,3,3-trifluoropropene (a fire extinguisher agent), on reproductive performance and 

development in Crl:CD (SD) rats following inhalation administration. Three groups of ten

male and ten female rats received 2-bromo-3,3,3-trifluoropropene by inhalation at target 

exposure levels of 200, 500 or 3000 ppm, 7 days per week, 6 hours per day. The adults were 

treated daily for a minimum of 15 days before pairing until Day 9 after the birth of the F1 

generation, and were sacrificed on Day 10 after the birth of the F1 generation. Females were 

untreated from Day 20 after mating until Day 4 after birth inclusive. A similarly constituted 

Control group received air only for the same duration. 

The F1 generation received no direct administration of the test substance; any exposure was 

in utero or via the milk.

During the study, clinical condition, bodyweight, food consumption, macroscopic and 

microscopic pathology investigations were undertaken in all adults. Oestrous cycles and 

gestation length were assessed and parturition observations were performed for F0 females. 

Organ weights were recorded for F0 males only. The clinical condition, litter size and 

survival, sex ratio and bodyweight of all offspring were assessed. 

Results

The achieved chamber concentrations of 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.

At 2900 ppm, treatment related findings included clinical signs during exposure of

underactivity, unresponsiveness, piloerection and partially closed eyelids, occasionally in 

females these lasted after completion of exposure, and included hunched posture, but not at 

the end of the day; pale teeth in all males and the majority of females and slightly higher 

incidence of hairloss in males (confirmed at macroscopic examination); lower bodyweight 

gain of males throughout and females prior to pairing and during gestation; lower food 

intake; increased water intake for females; longer oestrous cycles (6 days or longer) with

more females showing irregular cycles or being acyclic; reduced fertility (6/10 paired females 

were pregnant); longer pre-coital interval; at mating, fewer copulation 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, with only one female littering and the offspring subsequently dying; 

effects on sperm analysis (reduced percentage progressively motile sperm, sperm velocity, 

increased BCF, reduced sperm count in the cauda epididymis, increase in abnormal sperm –

breakages and abnormal head shape); reduced prostate, epididymides, seminal vesicle and 

pituitary weights; small prostate macroscopically in all males; in the majority of males and 

some females, spleen capsular thickening and adhesions noted macroscopically and 

confirmed microscopically as capsular/subcapsular inflammation/capsular thickening and /or 

adhesions/inflammation/fibrosis and reduced size of corpora lutea in the ovaries.

At 505 ppm, treatment related findings included clinical signs during exposure of

underactivity and piloerection and occasionally unresponsiveness and partially closed eyelids, 

occasionally in females some of these lasted after completion of exposure but not at the end 
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of the day; pale teeth in some males and females in the last week of treatment; lower 

bodyweight gain of males throughout and slightly lower bodyweight gain of females during 

gestation; slight bodyweight loss for females in lactation; lower food intake; increased water 

intake for females prior to pairing and during gestation, with lower water intake during 

lactation; longer oestrous cycles (6 days or longer) in 5/10 females with more females 

showing irregular cycles or being acyclic; slightly longer pre-coital interval; at mating, fewer 

copulation plugs and lower estimates of sperm in the vaginal smear; extended duration of 

gestation (23-25 days); slightly lower implantation counts; lower post implantation survival, 

live birth index and live litter size on Day 1; clinical signs in offspring of cold to touch, unfed 

and reduced activity necessitating sacrifice along with an increased incidence of dead 

offspring resulting in only 4/9 litters surviving to Day 10 post partum; effects on sperm 

analysis (reduced sperm velocity, increase in abnormal sperm – breakages and abnormal head 

shape); reduced prostate, epididymides, seminal vesicle and pituitary weights; small prostate 

macroscopically in the majority of males; and spleen capsular thickening and adhesions noted 

macroscopically in the majority of males and occasional females and confirmed 

microscopically as capsular/subcapsular inflammation/capsular thickening and /or 

adhesions/inflammation/fibrosis.

At 198 ppm, slight effects on clinical signs during exposure, bodyweight, food and water 

intake were considered not adverse. Of uncertain significance was a slightly lower sperm

count noted in the vaginal smear on the day of mating, but not statistically significantly 

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 slightly lower 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 viability indices (54 

and 82%), and these differences contributed to lower total and live litter sizes than the 

Controls. Although only a few litters were involved, the reduction in total and live litter size 

on Day 1 of lactation formed part of a dose related trend and thus a relationship to treatment 

was considered likely.

Conclusion

Exposure of 2-bromo-3,3,3-trifluoropropene to rats in this screening study to assess the 

influence on reproductive performance and development following administration by

inhalation at concentrations of 198, 505 or 2900 ppm was associated with adverse effects on 

male and female toxicity, and reproductive performance and development at 505 and 2900 

ppm.

Exposure at 505 and 2900 ppm produced effects on oestrous cycles, fertility (2900 ppm

only), extended duration of gestation, sperm counts and motility, implantation counts, 

reduced in utero and post partum survival, in the presence of signs of adult toxicity. In 

addition, at 2900 ppm, reduced size of corpora lutea in the ovaries was noted.

Exposure at 198 ppm was associated with slight effects on adult toxicity, 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 viability 

indices leading to lower litter size) as seen at 505 ppm and above.
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Histopathological examination 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. 

Within this screening study, the no adverse effect level (NOAEL) for reproductive 

performance and development was considered to lie below 198 ppm. 
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1. Introduction

1.1 Objective

The objective of this study was an initial screening assessment of the influence of

2-bromo-3,3,3-trifluoropropene, an experimental agent under development to replace halons 

as a fire extinguishing agent, on reproductive performance and development following

administration by inhalation for a minimum of 15 days before pairing until Day 9 after the 

birth of the F1 generation with sacrifice on Day 10 after the birth of the F1 generation.

1.2 Regulatory compliance

The study was designed to meet the requirements of the following guidelines:

Organisation for Economic Co-operation and Development: Testing of Chemicals 

(Guideline 421; Reproduction/Developmental Toxicity Screening Test; 27 July 

1994).

The study was conducted in accordance with the requirements of current, internationally

recognised Good Laboratory Practice Standards, and the applicable sections of the United 

Kingdom Animals (Scientific Procedures) Act 1986.

1.3 Test system

The rat was chosen as the test species because of the requirement for a rodent species by 

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

available at this laboratory. 

1.4 Route of administration

The inhalation route of administration was chosen to simulate the conditions 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 of 500 ppm to 20000 ppm, with excessive body weight effects at 10000 ppm 

and above. Respiratory tract pathology was noted at target exposure levels of 1200 ppm and 

above.

It was considered suitable for the target exposure levels of this reproductive study to replicate 

that of the 90 day rat study (Huntingdon Life Sciences Report Number WAG0014). The high 

exposure concentration, 3000 ppm, was selected to allow assessment of reproductive effects 

at an exposure concentration anticipated to produce evidence of systemic toxicity. Lower 

exposure concentrations were chosen to allow assessment of any effects observed.
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2. Experimental Procedure

2.1 Study schedule and structure

2.1.1 Duration of treatment

The test substance, 2-bromo-3,3,3-trifluoropropene, was administered to F0 adults for a 

minimum of 15 days before pairing until Day 9 after the birth of the F1 generation. The F0 

female adults were killed on Day 10 after the birth of the F1 generation and the F0 males 

killed after the start of the F0 female kills. F0 females were untreated from Day 20 after 

mating to Day 4 after birth. 

Animals of the F1 generation were not dosed, but may have been exposed to the test 

substance or metabolites in utero or through the milk.

2.1.2 Time schedule

Study initiation:

(Protocol signed by Study Director)

09 July 2012

Experimental start date:

(Animal arrival)

12 July 2012

Treatment commenced: 19 July 2012

Pairing commenced: 02 August 2012

Necropsy completed:

F0 Males

F0 Females and F1 animals

07 September 2012

13 September 2012

Experimental completion date:

(Pathology) 

13 November 2012

Study completion: When the Study Director 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 

lactation

Dams not exposed 

Day 20 post coitum 

to Day 4 post-partum 

inclusive
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2.1.4 Identity of treatment groups

The study consisted of one Control and three treated groups of rats, identified as follows:

Group Treatment Target 

exposure 

level (ppm)

Number of animals Animal numbers

Male Female Male Female

1 Control 0 10 10 201-210 241-250

2 2-bromo-3,3,3-

trifluoropropene

200 10 10 211-220 251-260

3 2-bromo-3,3,3-

trifluoropropene

500 10 10 221-230 261-270

4 2-bromo-3,3,3-

trifluoropropene

3000 10 10 231-240 271-280

The F1 generation received no direct administration of the test substance, 

2-bromo-3,3,3-trifluoropropene. Any exposure to the test substance or metabolites was 

through the mother to the offspring in utero and/or through the milk, or from transfer from 

the mother’s fur following exposure from Day 5 after birth.

2.2 Test substance and formulation

2.2.1 Test substance

Information 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 following information is given in summary: 

Identification: 2-bromo-3,3,3-trifluoropropene (stabilized)

Action: Fire extinguisher agent

Description: Clear liquid with a slight yellow hint

Storage conditions: At ambient temperature

Supplier: Sponsor

Batch number: WNCC-BTP111128

Date of receipt: 02 March 2012

Expiry date: 12 January 2013

Purity: 99.9% excluding stabilisers
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Stabilisers:

The Sponsor was responsible for the characterisation of the test substance and the 

documentation of the methods of synthesis, fabrication or derivation and stability.

2.2.2 Control

Group 1 (Control) animals were exposed to air only.

2.3 Animal management

2.3.1 Animal supply, acclimatisation and allocation

Crl:CD (SD) rats (a total of 45 males and 45 females) were received from Charles River (UK) 

Ltd. They were delivered at approximately 49 to 56 days of age and within a weight range of 

250 to 300 g for males and 200 to 230 g for females.

On arrival, the animals were removed from 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 with 

the procedure being repeated until each cage held the appropriate number of animals. Each 

sex was allocated separately.

The day after arrival a representative sample of animals were weighed and their clinical 

condition was satisfactory; based on this the animals were deemed acceptable for use on the 

study. On Day 1 (before dosing) variations in group bodyweights were confirmed to be 

within ± 20% of the mean weight for each sex. 

Each adult animal was assigned a number and identified uniquely within the study by a tail 

tattoo. Each cage label was colour-coded according to group and was numbered uniquely 

with cage and study number, as well as the identity of the occupant(s).

The adult animals were allowed to acclimatise to the conditions described below for 7 days

before treatment commenced. For those animals selected for this study, their age at the start 

of treatment was 56 to 63 days and their bodyweights were in the range of 303 to 349 g for 

males and 204 to 247 g for females.

The offspring were numbered individually within each litter on Day 1 of age, using a toe 

tattoo.

The spare animals were removed from the study room after treatment commenced.

2.3.2 Animal housing, diet and water supply

Animals were housed inside a restricted access rodent facility (Building Y14, Room 010). 

The facility was designed and operated to minimise the entry of external biological and 
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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 filtered fresh air, which was passed to atmosphere and not re-circulated. The temperature 

and relative humidity controls were maintained within the range of 19 to 25C and 40 to 70% 

respectively. Although conditions were occasionally outside the indicated ranges, these 

deviations were minor and/or of short duration and were not considered to have influenced 

the health of the animals and/or the outcome of the study. Artificial lighting was controlled 

to give a cycle of 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 ventilation system, or temperature limits 

were exceeded. A stand-by electricity supply was available to be automatically 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 Polycarbonate Solid polycarbonate

Pairing 1 : 1 Polycarbonate Stainless steel grid

Males after mating Up to 5 - Polycarbonate Solid polycarbonate

Gestation - 1 Polycarbonate Solid polycarbonate

Lactation - 1 (+ litter) Polycarbonate Solid polycarbonate

The cages were distributed on the racking to equalise, as far as possible, environmental 

influences amongst the groups. Wood based bedding was used in cages with 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 mating. IMS Nestlets were provided from Day 15 after mating and throughout 

parturition and lactation. Cages, cage-trays, food 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 pairing and lactation), and were 

replaced when necessary. Each cage of animals was provided with a plastic shelter for 

environmental enrichment (except during pairing, 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 antibiotic or other chemotherapeutic or prophylactic agent.

Potable water taken from the public supply was freely available via polycarbonate or 

polypropylene bottles fitted with sipper tubes.

Each batch of diet was analysed routinely by the supplier for various nutritional components 

and chemical and microbiological contaminants. Supplier's analytical certificates were 

scrutinised and approved before any batch of diet was released for use. The quality of the 
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water supply is governed by regulations published by the Department for Environment, Food 

and Rural Affairs. Certificates of analysis were received routinely from the water supplier, 

the bedding supplier and the suppliers of the Aspen chew blocks, IMS Nestlets and plastic 

shelters. Since the results of these various analyses did not provide evidence of 

contamination 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 generation. Animals of the F1 generation were not treated 

directly 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 whole body exposure systems. Animals in 

the treated groups were exposed to aerosols generated from 2-bromo-3,3,3-trifluoropropene 

at target exposure levels of 200, 500 and 3000 ppm. Control animals were exposed to air 

only, using the same dosing procedure. The relevant treatment was administered once daily 

for a period of 6-hours, seven days per week. Exposure of pregnant females was not 

undertaken on and after Day 20 after mating and not reintroduced until Day 4 post partum. 

The test atmosphere generation, administration, sampling and analysis and chamber 

environment, was 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 activities relating to the day by day running and maintenance 

of the study within the animal unit as well as to the group observations and examinations 

outlined in this experimental procedure were recorded in the Study Daybook. In addition, 

observations relating 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-health or reaction to 

treatment. Cages and cage-trays were inspected daily for evidence of ill-health amongst the 

occupant(s). Any deviation from normal was recorded at the time in respect of nature and 

severity, date and time of onset, duration and progress of the observed condition, as 

appropriate. 

In addition, a more detailed physical examination was performed once each week on each F0 

animal (females only until mating) and for F0 females on Days 0, 7, 14 and 20 after mating 

and Days 1, 5, and 9 of lactation to monitor general health. 

Daily during the first week of treatment, twice weekly during Weeks 2 to 4 and weekly 

thereafter for all adult animals (females only until mating) and, for females, on Days 0, 7, 14 
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and 19 after mating and Days 5 and 9 of lactation, detailed observations were recorded at the 

following times in relation to dose administration:

- Pre exposure observation

- Observations during exposure (limited to animals visible in the exposure 

chamber), recorded on a cage basis at approximately 2 and 4 hours after start of 

exposure

- Immediately after dosing on return of the animal to its cage

- As late as possible in the working day

For the observations during exposure, all animals in each group have been given any clinical 

signs observed as only a few animals could be observed.

2.4.2 Mortality

Debilitated animals were observed carefully and animals were killed for reasons of animal 

welfare where necessary. A complete necropsy was performed in all cases for animals killed 

or found dead, as described in the Necropsy section below. 

2.4.3 Bodyweight

The weight of each adult was recorded during acclimatisation, on the day that treatment 

commenced (Week 0), twice weekly thereafter (females only until mating) and before 

necropsy.

The weight of each F0 female was also recorded on Days 0, 3, 7, 10, 14, 17 and 20 after 

mating and on Days 1, 5 and 10 of lactation.

2.4.4 Food consumption

The weight of food supplied to each cage, that remaining and an estimate of any spilled was 

recorded on a weekly basis (from the first day of treatment until 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 acclimatisation of the males to the mating cages, the males were 

moved to the individual mating 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 for males in 

Week 2. For tabulated data the food consumption has been adjusted to provide a 

representative value for the entire week.

For each F0 female, the weight of food supplied, that remained and an estimate 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 consumption (g/rat/day) 

was calculated for each animal.

2.4.5 Water consumption 

Water consumption was recorded by weight (over a 3 day period on each occasion) on a daily

basis from 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.
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For each F0 female, water consumption was also recorded daily from Day 0 after mating and 

from Day 1 of lactation, and has been tabulated for the periods Days 0-2, 3-5, 6-8, 9-11, 12-

15, 16-17 and 18-19 after mating and Days 1-3 and 7-9 of lactation. 

2.4.6 Oestrous cycles

For 15 days before pairing, daily vaginal smears were taken from all females, using cotton 

swabs moistened with saline. The smears were subsequently examined to establish the 

duration and regularity of the oestrus cycle. After pairing with the male, smearing was 

continued using pipette lavage, until evidence of mating was observed.

2.4.7 Mating procedure

Following a minimum of two weeks of treatment, males and females were paired on a 

one-to-one basis from within the same treatment group for a period of up to two weeks.

Each morning following pairing, the trays beneath the cages were checked for ejected 

copulation plugs and a vaginal smear was prepared from each female and examined for the 

presence of spermatozoa and the stage of the oestrus cycle. The day on which evidence of 

mating was found was designated Day 0 of gestation. Once mating occurred, the males and 

females were separated and smearing was discontinued.

The pre-coital interval was calculated for each female as the time elapsing between initial 

pairing and detection of mating.

2.4.8 Parturition observations and gestation length

From Day 20 after mating, females were inspected three times daily for evidence of

parturition. The progress and completion of parturition was monitored, numbers of live and 

dead offspring were recorded and any difficulties observed were noted.

The duration of gestation was calculated as the time elapsing between the detection of mating 

and commencement of parturition. Half a day was subtracted in cases where parturition 

commenced overnight.

Females were sacrificed on Day 25 after mating if parturition was not complete or had not

started.

2.4.9 Records made during littering phase

All litters were examined at approximately 24 hours after birth (Day 1 of age) and then daily

thereafter. The records maintained were as follows:

Clinical signs: Daily records were maintained for evidence of ill health 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 mortality 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.
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Bodyweight: Individual offspring bodyweights 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 majority 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 after mating. 

Females whose litter died or was sacrificed before Day 10 of lactation were killed on the day

the last offspring died. 

Offspring were killed on Day 10 of age, unless they had shown evidence of ill health.

2.5.2 Method of kill

All adult animals were killed by carbon dioxide asphyxiation. Offspring were killed by

intraperitoneal injection of sodium pentobarbitone. 

The sequence in which the animals were killed after completion of the study was selected to 

allow satisfactory inter-group comparison.

2.5.3 Macroscopic pathology

All adult animals were subject to a detailed necropsy, which involved the following:

After a review of the history of each animal, a full macroscopic examination of the 

tissues was performed. All external features and orifices were examined visually. 

After ventral mid-line incision, the neck and associated tissues and the thoracic, 

abdominal and pelvic cavities and their viscera were exposed and examined in situ. 

Any abnormal position, morphology or interaction was recorded. External and cut 

surfaces of the organs and tissues were examined as appropriate. Any abnormality 

in the appearance or size of any organ and tissue was recorded and the required 

tissue samples preserved in appropriate fixative.

The requisite organs were weighed and external and cut surfaces of the organs and 

tissues were examined as appropriate. Any abnormality in the appearance or size of 

any organ and tissue was recorded and the required tissue samples preserved in 

appropriate fixative.

For F0 females, the numbers of implantation sites in each uterine horn was counted.

For females failing to produce a viable litter, the number of uterine implantation 

sites was re-checked after staining with ammonium sulphide (modification of the 

Salewski staining technique (Salewski, E, 1964)). 

Females whose litter died or was sacrificed before Day 10 of lactation were also examined 

for mammary tissue appearance. 

For offspring surviving to scheduled termination, a careful external examination was 

performed for gross abnormalities and externally normal offspring were discarded without 
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internal examination. Externally abnormal offspring were internally examined and any 

abnormal tissues were retained in an appropriate fixative.

Additionally the following procedures were applicable:

Premature deaths: Missing offspring and those grossly autolysed or grossly

cannibalised could not be examined. All other offspring dying or sacrificed before 

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 before disposal of the carcass.

2.5.4 Sperm analysis

Immediately after scheduled sacrifice of each F0 generation male, the left vas deferens, 

epididymis and testis was removed and the epididymis and testis were weighed. 

The following tests were performed:

Sperm motility: A sample of sperm was expressed from 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 action and where possible at least 200 sperm per animal 

analysed using the Hamilton Thorne IVOS Computer Assisted Sperm Analyser 

(CASA) version 12.3d. The percentages of motile and progressively motile sperm 

and sperm motion 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 

with nigrosine and eosin an air-dried smear was prepared and examined by light 

microscopy for 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 observations were reported.

Sperm count: The left cauda epididymis of each male was weighed and then frozen. 

After thawing, 10 mL of a mixture of 0.9% saline, 0.01% merthiolate and 0.05% 

Triton X-100 (SMT) was added and the cauda epididymis was homogenised for at 

least one minute. An aliquot of this mixture was added to a pre-prepared IDENT 

stain tube before being assessed for sperm count using CASA. The concentration 

(Million/g) and total number of sperm were reported.

Homogenisation-resistant spermatids count: The left testis of each male was 

frozen. After thawing, each testis was homogenised 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 homogenisation-resistant spermatid count using 

CASA. The concentration (Million/g) and total number of spermatids were 

reported.

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. 
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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 velocity, this is the average velocity of the smoothed cell

path.

VSL - the progressive or straight line velocity, this is the average velocity measured 

in a straight line from the beginning to the end of the track.

VCL - the curvilinear velocity or track speed, this is the average velocity measured 

over the actual point to point track followed by the cell.

ALH - amplitude 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 with 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 straight line.

LIN -linearity, 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 minor to major axis of all sperm heads.

Area - the average size of all sperm heads.

Rapid - the fraction of cells moving with VAP > progressive minimum VAP.

Medium - the fraction of cells moving with slow VAP cut-off < VAP < progressive 

minimum VAP.

Slow - the fraction of cells moving with VAP< slow VAP cut-off or VSL < slow 

VSL cut-off.

Static - the fraction of all cells which are not moving at all.

2.5.5 Organ weights

The following organs, taken from each male, were dissected free of adjacent fat and other 

contiguous tissue and the weights recorded:

Epididymides (L&R)

Testes (L&R)

Pituitary

Prostrate

Seminal vesicles

L&R Bilateral organs weighed individually

Organ weights were also adjusted for terminal bodyweight, using the weight recorded before 

necropsy.
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2.5.6 Fixation

Testes and epididymides were initially fixed in modified Davidson's fluid prior to transfer to 

10% neutral buffered formalin. Samples (or the whole) of the other tissues listed below from 

all animals were preserved in 10% neutral buffered formalin:

For adult animals:

Abnormalities Prostate

Epididymides+ Seminal vesicles#

Mammary area - caudal†# Testes+

Ovaries Uterus with cervix and oviducts#

Pituitary‡# Vagina#

# Not examined histologically

† Females with total litter loss only

‡ For males with suspect fertility only

+ Only right fixed and subject to histology and pathology; left organ used for sperm analysis

Samples of any abnormal tissues were also retained and processed for examination. In those 

cases where a lesion was not clearly delineated, contiguous tissue was fixed with the grossly 

affected region and sectioned as appropriate. 

Samples of the mammary area (from females with total litter loss only), pituitary (from males 

of suspect fertility only), uterus with cervix and oviducts and vagina were not examined 

histologically, but are retained against any future requirement for microscopic examination.

2.5.7 Histology

For those animals specified in the Pathology section, the relevant tissues were subject to 

histological processing.

Tissue samples were dehydrated, embedded in paraffin wax, sectioned at approximately four 

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 following regions:

Epididymides - caput, corpus and cauda

Ovaries - qualitative evaluation of one section from each ovary

For bilateral organs, sections of both organs were prepared. A single section was prepared 

from each of the remaining tissues required for microscopic pathology.
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2.6 Pathology

2.6.1 Light microscopy

Microscopic examination was performed as follows:

All tissues preserved for examination (as specified above) were examined for all 

animals of Groups 1 (Control) and 4 (2900 ppm) sacrificed on completion of the 

scheduled treatment period and for all animals killed or dying during the study.

Tissues reported at macroscopic examination as being grossly abnormal were 

examined for all animals in line with current practice (incisors or skin showing hair 

loss is not examined).

The following 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 severity grade. In the latter case one 

of the following five grades was used - minimal, slight, moderate, marked or severe. A 

reviewing pathologist undertook a peer review of the microscopic findings.

For the assessment of the ovaries, one mid-line section of each ovary was examined for the 

presence of primordial follicles, growing follicles and corpora lutea. 

For the assessment of the right testis, a detailed qualitative examination was made, taking into 

account the tubular stages of the spermatogenic cycle. The examination was conducted in 

order to identify treatment related effects such as missing germ cell layers or types, retained 

spermatids, multinucleate or apoptotic germ cells and sloughing of spermatogenic cells into 

the lumen. Any cell- or stage-specificity of testicular findings was noted.

For the assessment of the epididymides, leukocyte infiltration, change in prevalence in cell 

types, aberrant cell types and phagocytosis of sperm was examined. 

2.7 Computer systems

The computer systems with version numbers that were used on this study to acquire and 

quantify 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, 

bodyweights, food consumption, water consumption, organ weights, 

macropathology and histopathology, statistics.

Quasar (version 1.3) and SAS (version 9.1.3) in-house statistical analysis packages

used for bodyweight, food consumption and organ weight statistics.

Sample Registry System (version 1.0) used for Formulation and Inhalation Analysis 

data.

SAS (version 9.1.3) in house statistical analysis package used for sex ratio statistics. 
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StarTox (version 3.2) in-house statistical analysis package used for litter size and 

survival indices statistics.

StatXact 3 statistical analysis package used for mating performance and fertility and 

oestrous cycles and pre-coital interval statistics.

Waters Empower 2, Build 2154, Feature Release 4, Service Pack D and used for 

Formulation and Inhalation Analysis.

Xybion Path/Tox System (version 4.2.2) used for clinical signs, dosing signs, 

bodyweights, food consumption, water consumption, organ weights, 

macropathology and histopathology.

Xybion Pristima (version 6.3.2) used for Pharmacy test substance management.

2.8 Data treatment

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 deviations) presented in this report were 

calculated from computer-stored individual raw data. The summary statistics and the 

individual data were stored in the computer to a certain number of decimal places, different 

for each parameter. For presentation purposes, however, they were usually rounded to fewer 

places. It is, therefore, not generally possible to reproduce the presented means and standard 

deviations exactly using the presented individual data.

The codes in the appendices have the following meaning: 

T Scheduled kill; fertility 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 sites visible

TLL Total litter loss

U Unilateral pregnancy

Within the context of this study it 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 following abbreviations are used:

M Male

F Female

N/n Number contributing to the mean (normally the number animals/litters)

SD Standard deviation
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2.8.1 Signs

Signs are considered in two parts: observations in association with treatment, classified as 

“dosing signs” and extended changes in condition, classified as “clinical signs”. 

Signs considered to indicate a reaction to treatment are presented for each animal that shows 

signs, providing detail of type 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 different phases 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 of 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 weight changes of individual animals surviving the specified period. 

Bodyweight was plotted graphically with respect to the first day of the relevant period.

2.8.3 Food consumption

During the pre-pairing periods of the study where animals were housed 5 per cage, values 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 acclimatisation of the males to the mating cages, the 

males were moved to the individual mating 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 for 

males in Week 2. For tabulated data the food consumption has been adjusted to provide a 

representative value for the entire week.

During the gestation and lactation phases, where females were individually housed, group 

mean values and SD were calculated from individual values for 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 based on the numbers of 

females completing each phase. 

Mean overall values for food consumption before pairing were calculated from the weekly 

group mean values presented. 

2.8.4 Water consumption 

Water consumption was calculated from measurements of initial and final weights (g) of the 

water bottle and contents for each cage (it was assumed that 1 mL of water weighed 1 g).
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2.8.5 Oestrous cycles

The percentage females showing the following classifications of oestrous cycles before 

pairing are presented: 

Regular: All observed cycles of 4 or 5 days

Irregular: At least one cycle of 2, 3 or 6 to 10 days

Acyclic: At least 10 days without oestrus (beginning before pairing).

2.8.6 Pre-coital interval 

Individual intervals were tabulated for the time elapsing between initial pairing and mating. 

Percentage of females with pre-coital intervals 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 values were calculated for males and females 

separately for the following:

Percentage mating =

Number animals mating x 100 Animals paired

Conception rate (%) =

Number animals achieving pregnancy x 100 Animals mated

Fertility 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 parturition 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.
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2.8.10 Survival indices

The following were calculated for each litter:

Post-implantation 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 offspring 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 offspring in each litter was calculated at Day 1, and for live offspring 

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 following:

Motility (vas deferens) The percentage of motile and progressively motile sperm

and a detailed assessment of motility were reported. 

Morphology The number and percentages of normal and abnormal 

sperm were reported. A summary of the types of

abnormalities seen was made because there was evidence 

of an effect on the percentage normal sperm.

Count (Testis and Cauda 

epididymis)

The sperm concentration (Million/g) and total number 

were reported.

Group mean values were calculated from individual values.

2.8.13 Organ weights

The abbreviations used in the Appendix have the following meanings: 

Lt Left

Rt Right
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2.8.14 Pathology

The abbreviations used in the Appendix have the following meanings: 

Fac. Con. Factors contributing

Lt. Left

Rt. Right

Tissues which could not be examined are specified in the Appendix. The absence of a 

comment for a tissue scheduled for examination therefore indicates that the tissue was 

considered to be normal. In all tabular presentations of data the tissues specified in the 

protocol for histopathological examination precede other tissues.

For offspring, findings were presented on an individual basis. 

2.8.15 Statistical analysis

All statistical analyses were carried out separately for males and females.

Data relating to food consumption during the pre-pairing 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 following data types were analysed at each timepoint separately:

Bodyweight, using absolute weights and gains over appropriate study periods

Food consumption, over appropriate study periods

Organ weights, both absolute and adjusted for terminal bodyweight

Litter data and survival indices

Mating performance and fertility

The following sequence of statistical tests was used for bodyweight, food consumption, organ 

weights, litter size and survival indices data:

A parametric analysis was performed if Bartlett's test for variance homogeneity 

(Bartlett 1937) was not significant at the 1% level. For 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, with the error mean square from the 

one-way analysis of variance, were made. For all other analyses the F1 approximate 

test was applied. This test is designed to detect significant departure from 

monotonicity 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 deviations of the observed 

means from the maximum likelihood means, calculated under a constraint of 

monotonicity with the usual error mean square, EMS. The null hypothesis is that the 

true means are monotonically ordered. The test statistic is F1 = NMS/EMS which 

can be compared with standard tables of the F-distribution with 1 and EMS degrees 

of freedom. If the F1 approximate test for monotonicity of dose-response was not 

significant at the 1% level, Williams' test for a monotonic trend was applied. If the 
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F1 approximate test was significant, suggesting that the dose-response was not 

monotone, Dunnett's test (Dunnett 1955, 1964) was performed instead.

A non-parametric analysis was performed if Bartlett's test was still significant at the 

1% level following both logarithmic and square-root transformations. For 

pre-treatment data, Kruskal-Wallis’ test (Kruskal and Wallis 1952, 1953) was used 

to test for any group differences. Where this was significant (p<0.05) inter group 

comparisons using Wilcoxon rank sum tests (Wilcoxon 1945) were made. For all 

other analyses the H1 approximate test, the non-parametric equivalent of the F1 test

described above, was applied. This test is designed to be used when the main test 

for comparison of the means is a non-parametric monotonic trend test, such as 

Shirley's test (Shirley 1977). The test statistic compares the non-monotonicity sums 

of squares, NRSS, for the deviations of the observed mean ranks from the maximum 

likelihood mean ranks with the non-parametric equivalent of the error sums of 

squares, ERSS = N(N+1)/12. The test statistic is H1 = NRSS/ERSS which can be 

compared to standard tables of the χ

2

-distribution with 1 degree of freedom. If the 

H1 approximate test for monotonicity of dose-response was not significant at the 1% 

level, Shirley's test for a monotonic trend was applied. If the H1 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 (Fisher 1973) were performed. Treatment groups were compared using 

pairwise comparisons 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 linear model with binomial errors, a logit link 

function and litter as a random effect (Lipsitz et al 1991). Each treated group was compared 

to control using a Wald chi-square test. For sex ratio, the numerator was Number of males, 

the denominator was Number of live fetuses.

For gestation length, sperm counts and copulation plugs an exact two-tailed Linear-by-linear

test (Cytel 1995), with equally spaced scores, was applied to all groups. If the test was 

statistically 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 statistically significant 

(p≥0.05). 

For the number of females with one or more ‘abnormal’ oestrous cycle length of ‘3 or less’ or 

‘5 or more’ days an exact one-tailed (upper-tail) Cochran-Armitage test (Cytel 1995) was 

applied to all groups. If the test was statistically 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 (Cytel

1995) was applied to all groups, using scores appropriate to the severity of the observation 

assuming 4 day cycles to be normal. The categories were scored as follows: a 4 day cycle 

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 statistically 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 statistically significant (p≥0.05).
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For pre-coital intervals an exact one-tailed (upper-tail) Linear-by-linear test (Cytel 1995) was

applied to all groups, using the following scores: a 1-4 day interval was scored as 1, a 5-8 day 

intervals was scored as 2, a 9-12 day interval was scored as 3 and a 13-14 day interval was 

scored as 4. If the test was statistically 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 statistically significant (p≥0.05).

For number mating, number conceiving, number fertile and number of live litters born an 

exact one-tailed (lower-tail) Cochran-Armitage test (Cytel 1995) was applied to all groups. If 

the test was statistically 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 

statistically significant (p≥0.05).

If the exact version of the Linear-by-linear test could not be calculated (due to the size of the 

table containing the data), then the asymptotic version 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 used on the tables that contain 

statistical results is given below.

l Data were log transformed for the statistical analysis.

Ca Treated groups compared to Control using Cochran-Armitage test

Du Treated groups compared to Control using Dunnett’s test.

Lt Treated groups compared to Control using Linear-by-linear association test

Sh Treated groups compared to Control using Shirley’s test.

Wa Treated groups compared to Control using Wald’s test

Wi Treated groups compared to Control using Williams’ test.

Codes placed above adjusted means indicate that the comparisons were based on adjusted 

means.

* p<0.05

** p<0.01

2.9 Quality assurance and archiving procedures

2.9.1 Quality assurance

Details of the Quality Assurance inspections and audits are presented on the Quality 

Assurance Statement.

2.9.2 Archives

Following completion 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 of 

Huntingdon Life Sciences. Types of sample and specimen which are unsuitable, by reason of 
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instability, for long term retention and archiving may be disposed of after the periods stated 

in Huntingdon Life Sciences Standard Operating Procedures.

A copy of the final report and all Quality Assurance inspection records will be retained 

indefinitely. All other appropriate specimens and records will be retained for a minimum 

period of one year from 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 following deviations 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 consumption, 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 pituitary, prostates and seminal vesicles (with coagulating gland) from all males 

were weighed, not just those of suspect fertility.

The sex ratio of litters was also calculated and reported on Day 5 post partum, not 

just Day 1 and Day 10.

Water consumption was presented daily for individual values. Water consumption 

was tabulated on a weekly basis 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 protocol. 

These deviations were considered to have not affected the integrity of the study.
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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 follows:

Group Treatment Chamber concentration 

(ppm)

Target Achieved

1 Control 0 -

2

2-bromo-3,3,3-

trifluoropropene 200 198

3

2-bromo-3,3,3-

trifluoropropene 500 505

4

2-bromo-3,3,3-

trifluoropropene 3000 2900

The achieved aerosol concentrations were 99, 101 and 97% of the target concentration for 

Groups 2, 3 and 4, respectively. Adjustments 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 Mortality and animal fates

Table 1

At 505 ppm, 5 females 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 condition, 

being cold with reduced activity and were killed with the dams. In one case, all pups that 

were born were found dead at the completion of parturition check (total litter loss pre-Day 1). 

At 2900 ppm, 2 females were sacrificed on Day 24 after mating. Female 276 had partially 

closed eyes, hunched posture and piloerection, and female 279 had piloerection and 

perigenital staining and was noted to be attempting parturition. Both females were pregnant. 

The single dam (female 273) with a live litter born at 2900 ppm was killed for reasons of 

animal welfare following total litter loss.

3.2.2 Signs

Table 2-3 and Appendices 1-4

Appendices 1 and 2 show signs noted at the physical examination. 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, for females, the incidence of 

individual signs associated with dosing as the study progressed prior to mating and through 

gestation and lactation.
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Pale teeth were noted in all males and 9/10 females at 2900 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). 

Hairloss was noted at a higher incidence than controls in males at 2900 ppm.

Occasionally, in some females at 2900 ppm, piloerection, hunched posture, and underactivity 

were noted at the physical examination, generally performed weekly. 

During the 6 hour exposures, the following signs were noted:

At 2900 ppm underactivity, unresponsiveness, piloerection and eyelids partially closed were 

noted throughout. In males these signs were not apparent after completion of exposures. In 

females some of 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).

At 505 ppm, underactivity and piloerection were noted throughout, and unresponsiveness and 

eyelids partially closed were noted on fewer occasions. In males the signs were not noted 

following exposure, but in females during gestation and lactation, underactivity and 

piloerection, and in females during gestation only, eyelids partially closed were noted after 

exposure but not at the end of the working day.

At 198 ppm underactivity, unresponsiveness, piloerection and partially closed eyelids were 

noted at a much reduced incidence than that seen at higher levels; only underactivity and 

piloerection were noted immediately after exposure during gestation only.

3.2.3 Bodyweight

Figures 1-5, Tables 4-7 and Appendices 5-8

Mean bodyweight gain of males over the treatment period was lower than the controls in all 

groups given the test article (gain Day 1-50: 77, 63 and 47% of control at 198, 505 and 

2900 ppm respectively). The effect was apparent from Week 1.

Mean bodyweight gain of females over the 2 week period prior to pairing was lower than the 

controls at 2900 ppm (gain Day 1-15: 19% of control). There were no effects of treatment on 

bodyweight gain of females over the 2 week period prior to pairing at 198 or 505 ppm.

Group mean bodyweight on Day 0 of gestation for females at 2900 ppm was lower than the 

controls, reflecting the difference established prior to pairing. Mean bodyweight gain during 

gestation was lower than the controls in females at 505 and 2900 ppm (gain Day 0-20: 83 and 

43% of control respectively). At 198 ppm, mean bodyweight gain was marginally lower than 

the controls during Days 0-20 (92% of control).

During lactation, at 505 ppm slight mean bodyweight loss occurred once treatment was 

reintroduced on Day 5 of lactation (2g loss compared to 20g gain in the control group), and as 

such overall bodyweight gain during lactation was lower than the controls (13% of control). 

At 198 ppm bodyweight gain during Days 1-10 of lactation was 59% that of the control. No 

litters survived at 2900 ppm and hence no adult female bodyweights were recorded.
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3.2.4 Food consumption

Tables 8-10 and Appendices 9-11

Mean food consumption 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 

food 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 198, 505 and 2900 ppm respectively.

During the gestation phase, food consumption of treated pregnant females remained low and 

at a similar magnitude to that noted prior to pairing for treated females compared with 

controls (Day 0-19: 90, 89 and 78% of control at 198, 505 and 2900 ppm).

During the lactation phase, food consumption was lower than controls for females at 198 and 

505 ppm (73 and 60% of control respectively). No litters survived at 2900 ppm and hence no 

food consumption data was generated for lactating females.

3.2.5 Water consumption

Tables 11-13 and Appendices 12-14

Water consumption was recorded for males and females from Week 2 of the treatment phase 

onwards following a visual increase in consumption noted for females in Week 1. 

Measured water consumption of females in Week 2 was 17, 59 and 72% higher than the 

controls at 198, 505 and 2900 ppm respectively.

During the gestation phase, water consumption was slightly 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), although the magnitude of the difference was less marked than that recorded prior to 

pairing.

During the lactation phase, water consumption, once treatment was reintroduced on Day 5, 

was slightly lower for females at 198 and 505 ppm compared with control.

There were no effects on water consumption of males in Weeks 2, 5, 6 or 7 (water 

consumption was not recorded during the 2 week pairing 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.

At 2900 ppm, only 1 female had regular cycle lengths, although the cycle lengths recorded 

for this female were all 5 days in duration. Three females also had irregular cycles (at least 

one cycle 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 

without oestrus). Assessment of cycle lengths showed, in females at 2900 ppm, a statistically 
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significant incidence of females with longer cycle lengths, and a statistically significant 

incidence of longer specific cycle lengths, with 46% of the total recorded cycle lengths being 

6 to 10 days, and 18% of the recorded cycle lengths being 11-20 days, compared with no 

similarly longer cycle lengths being observed in the control group. Only 33% of the total

recorded cycle lengths at 2900 ppm were regular (4 or 5 days) compared to all of those in the 

control group.

At 505 ppm, only 5/10 females had regular cycle lengths, although in 4 of these 5 animals the 

cycle lengths were all 5 days. Four females had one or more irregular cycles and one female 

was acyclic (with a period of 12 days without oestrous). Assessment of the cycle lengths 

showed a statistically significant incidence of females with longer cycle lengths, and a 

statistically significant incidence of longer specific cycle lengths, with 21% of recorded cycle 

lengths being 6 or more days long. Although 79% of the recorded cycle lengths were 

considered regular (compared to 100% in the control group), 61% were of 5 days duration 

compared to only 18% of those in the control group.

At 198 ppm, all females had regular cycle lengths (4 or 5 days), and there was thus 

considered to be no adverse effect. There was, however, a statistically significant shift 

towards the longer cycle length with 70% of all cycles being 5 days duration compared with 

only 18% of the control cycles. 

3.2.7 Pre-coital interval, mating performance and fertility

Tables 15-18 and Appendix 15

At 2900 ppm, only 7 of the pairings had evidence of mating and of these only 4 occurred in 

the first 4 days after pairing (the time considered to generally reflect a mating at the first 

oestrous possibility). However, review of the time of successful 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 was a statistically significant trend towards lower numbers of

copulation plugs at mating. Copulation plugs give an indication of the number of times that 

mating occurred. Only 29% of the matings had 4-6 recorded copulation plugs compared to 

80% of the controls; the remaining 71% of matings had 2 or 3 copulation plugs. Sperm count 

estimated at mating showed a slightly lower estimate of the presence of sperm in the vaginal 

smear taken on the day of mating, with the majority of smears having continuous few sperm 

or many scattered sperm compared to the solid masses of sperm seen more frequently in the 

control group. Of the 7 pairs of males and females showing evidence of mating, 6 pairings 

produced a pregnancy.

At 505 ppm all 10 pairings showed evidence of mating but 3 of these were in days 5-8 after 

pairing, not as would be usually expected in the first 4 days after pairing. A review of the 

time of successful mating showed that all but one females mated successfully at the first 

oestrous possibility. There was a statistically significant trend towards lower numbers of

copulation plugs at mating. Only 20% of the matings had 4-6 recorded copulation plugs 

compared to 80% of the controls; the remaining 80% of matings had 1, 2 or 3 copulation 

plugs. Sperm count estimated at mating showed a trend towards slightly lower estimates of

the presence of sperm in the vaginal smear taken on the day of mating, with the majority of

smears having continuous few sperm or many scattered sperm compared to solid masses of 

sperm as seen more frequently in the control group. Four of the matings had no sperm present 
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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.

At 198 ppm and in the control group, the majority of animals showed evidence of mating 

(pre-coital interval) in the first 4 days after pairing (i.e. at the first mating opportunity). With 

the exception of a trend towards a lower sperm count category in the vaginal smear on the 

day of successful mating, there were no effects on the number of copulation plugs, or mating 

performance. 

3.2.8 Gestation length and gestation index

Table 19 and Appendix 15

The duration of gestation in females from all treated groups was longer than the controls. 

In the control group, the females showed the typical pattern of a majority of gestation lengths 

of 22 days. Females at 198 ppm had gestation lengths of 23 days, and those at 505 ppm had 

gestation lengths of 23, 24 or 25 days. After 25 days of gestation only one female at 

2900 ppm had completed parturition.

The gestation index was reduced at 2900 ppm (only 17%) reflecting the single litter born 

from those mating in this group.

3.2.9 Sperm analysis

Tables 25-27 and Appendices 23-25

No adverse effects on percent motile sperm or testicular spermatid numbers were seen after 

treatment with 2-bromo-3,3,3-trifluoropropene at exposure levels up to 2900 ppm. 

Slight but statistically significant reductions in percent progressively motile sperm, velocities 

and percent medium sperm were observed after treatment at 2900 ppm. In addition a 

statistically significant increase in Beat Cross Frequency was seen at this level. Significant 

reductions in velocities were also apparent at the lower exposure levels of 198 and 505 ppm.

A slight but statistically significant reduction in number of sperm in the cauda epididymis 

was apparent after treatment at 2900 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 

were statistically significant at 505 and 2900 ppm. The corresponding increase in percent 

total abnormal sperm was significant at all exposure levels. Observed abnormalities were 

primarily breakages (including decapitate sperm) and abnormal head shape.

The increased Beat Cross Frequency suggests there was no reduction in vigour of the sperm 

but that their forward movement was affected, whilst the breakages suggest sperm fragility 

was increased.
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3.2.10 Organ weights

Table 28 and Appendix 26

At 2900 ppm, several organ weights were lower than the controls, notably prostate (52% of 

control), seminal vesicles (71% of control), epididymides (87% of control) and pituitary

(77% of control).

At 505 ppm, prostate (74% of control), seminal vesicles (70% of control) and pituitary (85% 

of control) weights were reduced.

At 198 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 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 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 examined 10 10 10 10

Spleen

Capsular thickening was seen in the 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
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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 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 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 consistent with the common 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-trifluoropropene 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.
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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 treatment were 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 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/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.

The histopathological examination of testes and epididymides from the F0 generation animals 

performed after scheduled treatment period revealed no findings attributable to treatment 

with the test article 2-bromo-3,3,3-trifluoropropene. 
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The seminiferous tubules were evaluated with respect to their stage in the spermatogenic 

cycle and the integrity of the various cell types present within the 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 findings to correlate with the organ weight 

variation 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 follicle counts.

3.3 F1 litter responses

3.3.1 Signs of offspring

Appendix 20

At 505 ppm the majority of offspring born were noted as being cold on Day 1, with those 

from dams where the litter was sacrificed early or died frequently 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 Appendices 16-18

At 2900 ppm the mean number of implantations in the pregnant females was reduced (9.5 

compared with 15.9 in the control group). Only one female produced a litter, in this female 

there was low post implantation survival (25%) and a low live birth index (33%) such that 

only one pup was alive on Day 1 post partum. This pup was sacrificed due to poor condition 

on Day 1 post partum. 

At 505 ppm the mean number of implantations was slightly lower than the controls. Post 

implantation survival, the live birth index and the viability index were all lower than the 

controls, resulting in a lower total and live litter size on Day 1 post partum, and lower litter 

size by Day 10 post partum (mean of 8.3 compared to 14.7 in the control group). In 

addition, a number of offspring were sacrificed due to poor condition including reduced 

activity and reduced body temperature. 

At 198 ppm there was a slightly lower mean number of implantations. Post implantation 

survival was slightly reduced which resulted in a lower total and live litter size on Day 1 

post partum. Group mean survival from 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

selective demise of the males.
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3.3.3 Bodyweight

Figure 5, Tables 23-24 and Appendix 19

There were no effects on mean male or female pup weight on Day 1 at 198 or 505 ppm. With

the longer duration of gestation at 198 and 505 ppm, it might be expected that pup weight 

would be higher, and this was the case at 198 ppm. Bodyweight gain from Day 1 to Day 10 

was slightly lower than the controls at 198 and 505 ppm.

There was only a single female pup weighed at 2900 ppm, the weight was slightly lower 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 

most 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.
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4. Discussion

Exposure of 2-bromo-3,3,3-trifluoropropene to rats was undertaken as an initial screening 

assessment of the influence on reproductive performance and development following 

administration by inhalation for a minimum of 15 days before pairing until Day 9 after the 

birth of the F1 generation, and were sacrificed on Day 10 after the birth of the F1 generation. 

Females were not exposed between Day 20 after mating and Day 4 of lactation inclusive.

All females exposed to 2900 ppm and half of those exposed to 505 ppm were killed before 

Day 10 of lactation as they failed to mate (sacrificed on Day 25 after the last paring), failed to 

litter (sacrificed on Day 25 after mating), lost 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 cycles 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 

pairings with evidence of mating, and on the day of mating, a reduced presence of copulation 

plugs or sperm within the vaginal smear. The fertility index was reduced at 2900 ppm, 

probably linked with the marked effect on oestrous cycles. There was also a reduced number 

of implantations evident at sacrifice.

Of females that showed evidence of mating, the delayed pre-coital interval is considered in 

part to reflect the reduction in regular oestrous cycles in these groups, as successful matings 

tended to occur at the first oestrous opportunity after pairing, 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 

complete parturition on Day 22 after mating. At 2900 ppm only one of the six pregnant 

females littered, on Day 25 after mating. Two other females were sacrificed on Day 24 after 

mating, the signs indicated this was possibly associated with stress involved in 

commencement of parturition. The remaining 3 females were sacrificed on Day 25 after 

mating as parturition 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 parturition 

and the pups born were eaten prior to being observed at the animal room checks. At

505 ppm 9 of the 10 pregnant females produced a litter, although in one case all pups were 

found to be dead at the Day 1 check. The duration of gestation, in females at 505 ppm, was 

extended to between 23 and 25 days.

At 2900 and 505 ppm, a lower number of implantations, coupled with reduced in utero 

survival and reduced post partum viability resulted in reduced number of offspring surviving 

to Day 10 post partum; this being a combination of all offspring 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, inactive or unfed. 

A reduction in the number of sperm in the cauda epididymis at 2900 ppm correlated with the 

lower caudal epididymal weight, although the number of sperm/g epididymis was unaffected 
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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 copulation. At 2900 ppm and 

generally at 505 ppm, a reduced percentage progressively motile sperm and sperm velocity

and an increased Beat Cross Frequency suggested that whilst there was no reduction in vigour 

of the sperm, their forward movement was affected, and an increase in abnormal sperm 

(mainly breakages) suggested that sperm fragility was increased.

The reduced bodyweight 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 

lower numbers of implantations. At 505 ppm 4 females and their offspring did not survive to 

Day 10 post partum.

The clinical signs noted during exposure, notably reduced activity and unresponsiveness, 

were not unexpected following 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 activity monitoring in Week 13 showed reduced low and high 

level activity at similar exposure levels on the morning after exposure. It is uncertain what 

effect, if any, the reduced activity had on the mating performance. 

Water intake was generally higher in females at 505 or 2900 ppm prior to pairing and 

throughout gestation. During lactation however, water consumption was lower. The 

significance of this is unclear but it 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 cycles or the 

prolonged gestation lengths.

The examination of the ovaries from the F0 generation females revealed the presence of 

histopathological changes. A reduction in size of corpora lutea was observed in the ovaries 

of four females exposed to 2900 ppm and one female exposed to 198 ppm. The corpora lutea 

were quite small and appeared to be not newly formed. As this change was not consistent 

with 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 with small corpora lutea were found to be non pregnant; since this finding was not 

apparent amongst the pregnant females in this group it is more 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 examination. 

This was consistent with a reduction in prostate weight seen with increasing exposure level. 

No microscopic correlate for the small prostates could be identified. A low incidence of 

inflammatory changes was observed in all groups including 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 capsular/subcapsular inflammation 

and/or capsular thickening and/or adhesions/inflammation/fibrosis seen at the microscopic 

examination and are likely to be related to treatment. However, as no controls and only 
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tissues with macroscopic changes were sampled and microscopically examined, conclusions 

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 examination was not further clarified as these tissues are not 

routinely examined microscopically. 

Full evaluation of the spleen capsular thickening and adhesions, pale incisors and hairloss are 

considered to be within the remit of the concurrent 13 week toxicity study in the rat at the 

same target exposure concentrations [WAG0014].

It was considered that the effects seen at 505 and 2900 ppm were considered adverse.

At 198 ppm some similar effects were apparent but the magnitude of the responses tended to 

be much lower and most likely not to be adverse. These not adverse effects included the 

effects on bodyweight, food intake, water intake, and probably the shift to a 5 day oestrous 

cycle length (which is still considered normal). There was a slightly lower sperm count noted 

in the vaginal smear on the day of copulation, although at termination the epididymal sperm 

count was not statistically significantly reduced, there was no effect on the epididymal weight 

and as such any adversity 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. 

At 198 ppm, the mean number of implantations was slightly lower than in 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 viability indices (54 and 82%), and 

these differences contributed to lower total and live litter sizes than in Controls. Although 

only a few litters were involved, the reduction in total and live litter size on Day 1 of lactation 

formed 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 cycles). However, which sex(es) contributed to the 

reductions in implantation counts and the pre and post natal survival of the offspring cannot 

be determined within the context of this screening study. 
Huntingdon Life Sciences

WAG0015

48

5. Conclusion

Exposure of 2-bromo-3,3,3-trifluoropropene to rats in this screening study to assess the 

influence on reproductive performance and development following administration by

inhalation at concentrations of 198, 505 or 2900 ppm was associated with adverse effects on 

male and female toxicity, and reproductive performance and development at 505 and 

2900 ppm.

Exposure at 505 and 2900 ppm produced effects on oestrous cycles, fertility (2900 ppm

only), extended duration of gestation, sperm counts and motility, implantation counts, 

reduced in utero and post partum survival, in the presence of signs of adult toxicity. In 

addition, at 2900 ppm, reduced size of corpora lutea in the ovaries was noted.

Exposure at 198 ppm was associated with slight effects on adult toxicity 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 viability 

indices leading to lower litter size) as seen at 505 ppm and above.

Within 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

49 

6. References 

6.1 Necropsy and histology 

Salewski E (1964). Farbemethode zum makroscopischen nachweis von implantationsstellen 

am uterus der Ratte. Naunyn - Schmiedebergs Arch. exp. Pathol. Pharmacol., 247, 367. 

6.2 Statistics 

Bartlett MS (1937) .Properties of sufficiency 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 procedure for comparing several treatments with 

a control. Journal of the American Statistical Association 50, 1096-1121. 

Dunnett CW (1964). New tables for multiple comparisons with a control. Biometrics 20, 

482-491. 

Fisher RA (1973). Statistical Methods for 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 Finding Study by Inhalation Administration to 

Rats for 2 Weeks. 

Huntingdon Life Sciences Study number: WAG0014 

2-bromo-3,3,3-trifluoropropene: Toxicity Study by Inhalation Administration to Rats for 13 

Weeks. 

Huntingdon Life Sciences Study number: WAG0024 

2-bromo-3,3,3-trifluoropropene: Dose Range Finding Study by Inhalation 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. 
50

Huntingdon Life Sciences

WAG0015

FIGURE 1

Bodyweight - group mean values for males - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Pairing commenced Day 15
51

Huntingdon Life Sciences

WAG0015

FIGURE 2

Bodyweight - group mean values for females before pairing - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900
52

Huntingdon Life Sciences

WAG0015

FIGURE 3

Bodyweight - group mean values for females during gestation - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

a Mean of females which littered

b Mean of all pregnant females
53

Huntingdon Life Sciences

WAG0015

FIGURE 4

Bodyweight - group mean values for females during lactation - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Treatment reintroduced Day 5 of lactation
54

Huntingdon Life Sciences

WAG0015

FIGURE 5

Bodyweight - group mean values for offspring - male animals - F1 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900
55

Huntingdon Life Sciences

WAG0015

FIGURE 5 - continued

Bodyweight - group mean values for offspring - female animals - F1 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900
56

Huntingdon Life Sciences

WAG0015

TABLE 1

Summary of adult performance

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Group: 1 2 3 4

Achieved concentration (ppm): 0 198 505 2900

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 (non-pregnant) 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 pre Day 1 of lactation 0 0 1 (266) 0

Live litter born 10 9 8 1

Total litter loss post-partum 0 0 4 (262

a

, 265, 267, 270) 1 (273)

Litter surviving to Day 10 post partum 10 9 4 0

( ) Animal number

a

Includes dam/pups killed for welfare reasons 
57

Huntingdon Life Sciences

WAG0015

TABLE 2 Request ID: 284416

Clinical signs - group distribution of observations for males - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

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 10
58

Huntingdon Life Sciences

WAG0015

TABLE 2 - continued Request ID: 284417

Clinical signs - group distribution of observations for females before pairing - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 Conformation 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 3
59

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Underactive 0 0 0 2

Body temperature Reduced 0 0 0 1

Build Conformation 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 Abnormal Colour, Brown, Head 0 1 2 5

Abnormal Colour, Brown, Muzzle 0 0 0 2
60

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Staining Abnormal Colour, Red, Perigenital 0 0 0 1

Teeth Abnormal Colour, Pale 0 0 3 6
61

Huntingdon Life Sciences

WAG0015

TABLE 2 - continued Request ID: 284419

Clinical signs - group distribution of observations for females during lactation - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 1
62

Huntingdon Life Sciences

WAG0015

TABLE 3 Request ID: 284420

Signs associated with dosing - group distribution of observations for males - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Underactive 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

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 sign
63

Huntingdon Life Sciences

WAG0015

TABLE 3 - continued Request ID: 284420

Signs associated with dosing - group distribution of observations for males - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 sign 
64

Huntingdon Life Sciences

WAG0015

TABLE 3 - continued Request ID: 284420

Signs associated with dosing - group distribution of observations for males - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

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 sign 
65

Huntingdon Life Sciences

WAG0015

TABLE 3 - continued Request ID: 284420

Signs associated with dosing - group distribution of observations for males - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 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

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)

Lachrymation 1 0 0 10 0

Eye (Right)

Lachrymation 1 0 0 10 0

Signs recorded during exposure on a group basis, all animals in the group assumed to have the same sign 
66

Huntingdon Life Sciences

WAG0015

TABLE 3 - continued Request ID: 284420

Signs associated with dosing - group distribution of observations for males - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 (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 sign 
67

Huntingdon Life Sciences

WAG0015

TABLE 3 - continued Request ID: 284420

Signs associated with dosing - group distribution of observations for males - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 sign 
68

Huntingdon Life Sciences

WAG0015

TABLE 3 - continued Request ID: 284421

Signs associated with dosing - group distribution of observations for females before pairing - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Underactive 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 the same sign
69

Huntingdon Life Sciences

WAG0015

TABLE 3 - continued Request ID: 284421

Signs associated with dosing - group distribution of observations for females before pairing - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 animals in the group assumed to have the same sign 
70

Huntingdon Life Sciences

WAG0015

TABLE 3 - continued Request ID: 284421

Signs associated with dosing - group distribution of observations for females before pairing - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

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 sign 
71

Huntingdon Life Sciences

WAG0015

TABLE 3 - continued Request ID: 284421

Signs associated with dosing - group distribution of observations for females before pairing - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 10 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)

Lachrymation 1 0 0 10 0

Signs recorded during exposure on a group basis, all animals in the group assumed to have the same sign 
72

Huntingdon Life Sciences

WAG0015

TABLE 3 - continued Request ID: 284421

Signs associated with dosing - group distribution of observations for females before pairing - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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)

Lachrymation 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

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

Signs recorded during exposure on a group basis, all animals in the group assumed to have the same sign 
73

Huntingdon Life Sciences

WAG0015

TABLE 3 - continued Request ID: 284421

Signs associated with dosing - group distribution of observations for females before pairing - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 sign
74

Huntingdon Life Sciences

WAG0015

TABLE 3 - continued Request ID: 284422

Signs associated with dosing - group distribution of observations for females after mating - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Underactive 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

Breathing

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

Signs recorded during exposure on a group basis, all animals in the group assumed to have the same sign
75

Huntingdon Life Sciences

WAG0015

TABLE 3 - continued Request ID: 284422

Signs associated with dosing - group distribution of observations for females after mating - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 sign
76

Huntingdon Life Sciences

WAG0015

TABLE 3 - continued Request ID: 284423

Signs associated with dosing - group distribution of observations for females during lactation - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Underactive 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 sign
77

Huntingdon Life Sciences

WAG0015

TABLE 4 Request ID: 284760

Bodyweight and bodyweight change - group mean values for males - F0 generation (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

1M 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 10 10 10

2M 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

3M 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

4M 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 15
78

Huntingdon Life Sciences

WAG0015

TABLE 4 - continued Request ID: 284760

Bodyweight and bodyweight change - group mean values for males - F0 generation (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

1M Mean 542 91 - 121 - 212 -

SD 33.0 15.4 19.9 27.9

N 10 10 10 10

2M Mean 483** 74 81 90** 74 164** 77

SD 52.0 30.6 17.4 44.0

N 10 10 10 10

3M Mean 455** 54** 60 79** 65 133** 63

SD 27.4 13.7 15.4 22.1

N 10 10 10 10

4M 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 15
79

Huntingdon Life Sciences

WAG0015

TABLE 5 Request ID: 284761

Bodyweight and bodyweight change - group mean values for females before pairing - F0 generation (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

1F Mean 202 225 237 239 245

SD 9.7 11.5 12.1 11.2 10.8

N 10 10 10 10 10

2F Mean 205 227 239 244 252

SD 6.2 5.7 9.0 9.3 10.9

N 10 10 10 10 10

3F Mean 202 225 232 236 244

SD 6.2 9.5 7.8 9.9 10.4

N 10 10 10 10 10

4F 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 15
80

Huntingdon Life Sciences

WAG0015

TABLE 5 - continued Request ID: 284761

Bodyweight and bodyweight change - group mean values for females before pairing - F0 generation (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Group Day Day Change As % of

/Sex 50 53 1-15 Control

Statistical test: Wi

1F Mean 20 -

SD 7.1

N 10

2F Mean 24 121

SD 10.3

N 10

3F Mean 19 95

SD 6.5

N 10

4F Mean 250 253 4** 19

SD 7.9 9.8 14.7

N 3 3 10

Pairing commenced Day 15
81

Huntingdon Life Sciences

WAG0015

TABLE 6 Request ID: 284780

Bodyweight and bodyweight change - group mean values for females during gestation - F0 generation (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure 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

1F 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

2F Mean 255 272 286 296 317 347 397 41 85 100 95

SD 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

3F 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

4Fa Mean 227 234 239 248 272 281 316 21 43 68 64

N 1 1 1 1 1 1 1 1 1

4Fb 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 which littered, not included in statistical evaluation

b Mean of all pregnant females, not included in statistical evaluation

Treatment withdrawn after Day 19 of gestation
82

Huntingdon Life Sciences

WAG0015

TABLE 6 - continued Request ID: 284780

Bodyweight and bodyweight change - group mean values for females during gestation - F0 generation (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Group Change As % of

/Sex 0-20 Control

Statistical test: Wi

1F Mean 154 -

SD 18.4

N 10

2F Mean 142 92

SD 27.7

N 9

3F Mean 128* 83

SD 16.3

N 9

4Fa Mean 89 58

N 1

4Fb Mean 66 43

SD 30.1

N 6

a Mean of females which littered, not included in statistical evaluation

b Mean of all pregnant females, not included in statistical evaluation

Treatment withdrawn after Day 19 of gestation
83

Huntingdon Life Sciences

WAG0015

TABLE 7 Request ID: 284763

Bodyweight and bodyweight change - group mean values for females during lactation - F0 generation (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

1F 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

2F 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

3F 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 lactation
84

Huntingdon Life Sciences

WAG0015

TABLE 8 Request ID: 284799

Food consumption - group mean values for males and females before pairing - F0 generation (g/animal/week)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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.
85

Huntingdon Life Sciences

WAG0015

TABLE 8 - continued Request ID: 284799

Food consumption - group mean values for males and females before pairing - F0 generation (g/animal/week)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

2F 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 2
86

Huntingdon Life Sciences

WAG0015

TABLE 9 Request ID: 284781

Food consumption - group mean values for females during gestation - F0 generation (g/animal/day)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 which littered, not included in statistical evaluation

b Mean of all pregnant females, not included in statistical evaluation
87

Huntingdon Life Sciences

WAG0015

TABLE 10 Request ID: 285999

Food consumption - group mean values for females during lactation - F0 generation (g/animal/day)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 lactation
88

Huntingdon Life Sciences

WAG0015

TABLE 11

Water consumption - group mean values for males and before pairing for females - F0 generation (mL/animal/day)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Group Animal 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 data 
89

Huntingdon Life Sciences

WAG0015

TABLE 11 - continued

Water consumption - group mean values for males and before pairing for females - F0 generation (mL/animal/day)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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/week
90

Huntingdon Life Sciences

WAG0015

TABLE 12

Water consumption - group mean values for females during gestation - F0 generation (mL/animal/day)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

1F 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

2F 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

3F 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

4Fa 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

4Fb 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 which littered, not included in statistical evaluation

b Mean of all pregnant females, not included in statistical evaluation
91

Huntingdon Life Sciences

WAG0015

TABLE 12 - continued

Water consumption - group mean values for females during gestation - F0 generation (mL/animal/day)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

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

1F 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

2F 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

3F 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

4Fa 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

4Fb 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 which littered, not included in statistical evaluation

b Mean of all pregnant females, not included in statistical evaluation
92

Huntingdon Life Sciences

WAG0015

TABLE 13

Water consumption - group mean values for females during lactation - F0 generation (mL/animal/day)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

1F 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

2F 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

3F 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 4
93

Huntingdon Life Sciences

WAG0015

TABLE 14

Oestrous cycles - incidence of females with specific cycle lengths - group values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Number

of Cycle 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 female may have cycles in more than one category)
94

Huntingdon Life Sciences

WAG0015

TABLE 14 - continued

Oestrous cycles - incidence of specific cycle lengths - group values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Number Number

of of Cycle 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 category
95

Huntingdon Life Sciences

WAG0015

TABLE 15

Pre-coital interval - group values- F0 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Number 

of Pre-coital interval (days)

Group animals1-45-89-12 13-14

Statistical test: Lt 1 10

n9100

(%) (90) (10)

2 10n9010

(%) (90) (10)

3 10n7300

(%) (70) (30)

47n4210

(%) (57) (29) (14)
96

Huntingdon Life Sciences

WAG0015

TABLE 16

Number of copulation plugs at mating - group values- F0 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Number

of Number of copulation plugs

Group animals123 4 -6

Statistical test: Lt 1 10

n0028

(%) (20) (80)

2 10n1117

(%) (10) (10) (10) (70)

3* 10n1162

(%) (10) (10) (60) (20)

4*7n0232

(%)(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 pairing
97

Huntingdon Life Sciences

WAG0015

TABLE 17

Sperm count estimates from vaginal smears at mating - group values- F0 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Number

of Sperm count category 



Group animals01234

Statistical test: Lt 1 10

n10126

(%) (10) (10) (20) (60)

2 10n03241

(%) (30) (20) (40) (10)

3 10n41122

(%) (40) (10) (10) (20) (20)

47n00241

(%) (29) (57) (14)

 0 No sperm

1 Occasional sperm

2 Continuous few sperm

3 Many scattered sperm

4 Solid masses of sperm
98

Huntingdon Life Sciences

WAG0015

TABLE 18

Mating performance and fertility - group values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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*

Statistical 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*
99

Huntingdon Life Sciences

WAG0015

TABLE 19

Gestation length and gestation index - group values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 23.5 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 pairing 
100

Huntingdon Life Sciences

WAG0015

TABLE 20

Litter size - group mean values for litters that survived to Day 10 of lactation - F1 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 Meana15.5 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

Meanb14.1 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 Meana9.5

n 6

Meanb12.0 3.0 1.0

n 1# 1 1

# Implantations only 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 applicable 
101

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TABLE 21

Offspring survival indices - group mean values for litters that survived to Day 10 of lactation - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 Meana

76.8* 91.3 78.2*

n444

Mean

b

56.5** 51.7** 62.6**

n 1095

4 Mean

a

25.0 33.3

n 1# 1#

Mean

b

4.2**

n6

# 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 applicable
102

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TABLE 22

Sex ratio - group mean values for litters that survived to Day 10 of lactation - F1 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 4
103

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TABLE 23

Bodyweight and bodyweight change - group mean values (g) for male offspring from litters surviving to Day 10 of lactation - F1 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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: 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 4
104

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TABLE 24 

Bodyweight and bodyweight change - group mean values (g) for female offspring from litters surviving to Day 10 of lactation - F1 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 Mean 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 evaluation
105

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TABLE 25

Sperm analysis - group mean values

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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) (millions/g) (million) (g) (millions/g) (million)

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 10
106

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TABLE 26

Sperm motion data - group mean values

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 10
107

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TABLE 27

Sperm morphology - group mean values

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Group Number Total number Normal Total abnormal Decapitate Head abnormal 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 Mean 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 0.4
108

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WAG0015

TABLE 28 Request ID: 284533

Organ weights - group mean unadjusted and adjusted values (g) for males - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Group

/Sex

Terminal 

Bodyweight

Epididymides Pituitary Prostate Seminal 

Vesicles

Testes

Unadjusted 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* 3.49
109

Huntingdon Life Sciences

WAG0015

TABLE 29 Request ID: 289375

Macropathology - group distribution of findings for males - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 0
110

Huntingdon Life Sciences

WAG0015

TABLE 29 - continued Request ID: 289377

Macropathology - group distribution of findings for decedent females - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 1
111

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 1
112

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Stomach

Cyst(s) 0 1 0 0

Liver

Pale 0 0 0 1
113

Huntingdon Life Sciences

WAG0015

TABLE 29 - continued Request ID: 289378

Macropathology - group distribution of findings for all females - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 2
114

Huntingdon Life Sciences

WAG0015

TABLE 29- continued Request ID: 289378

Macropathology - group distribution of findings for all females - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 1
115

Huntingdon Life Sciences

WAG0015

TABLE 30 Request ID: 289389

Histopathology - group distribution of findings for males - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (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 0
116

Huntingdon Life Sciences

WAG0015

TABLE 30 - continued Request ID: 289391

Histopathology - group distribution of findings for decedent females - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 1
117

Huntingdon Life Sciences

WAG0015

TABLE 30 - continued Request ID: 289390

Histopathology - group distribution of findings for females killed after scheduled treatment period - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 0
118

Huntingdon Life Sciences

WAG0015

TABLE 30 - continued Request ID: 289392

Histopathology - group distribution of findings for all females - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Capsular 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 Necrosis 0 0 0 1

Pelvic Dilatation 0 1 0 2

Stomach Number Examined: 0 1 0 0

Squamous Cyst - Limiting Ridge 0 1 0 0
119

Huntingdon Life Sciences

WAG0015

TABLE 31

Primordial ovarian follicle counts - group mean values (F0 generation)

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 10
120

Huntingdon Life Sciences

WAG0015

APPENDIX 1 Request ID: 284403

Clinical signs - individual observations for males - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 Terminal sacrifice

Only animals with observations are presented
121

Huntingdon Life Sciences

WAG0015

APPENDIX 1 - continued Request ID: 284403

Clinical signs - individual observations for males - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 presented
122

Huntingdon Life Sciences

WAG0015

APPENDIX 1 - continued Request ID: 284403

Clinical signs - individual observations for males - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 presented
123

Huntingdon Life Sciences

WAG0015

APPENDIX 2 Request ID: 284404

Clinical signs - individual observations for females - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Lactation Coat Hair loss, Forelimbs 9-10

245 T Pre-treatment No abnormalities detected

Treatment Behaviour Irritable 12

Vocalisation 12

Gestation No abnormalities detected

Lactation Coat Hair loss, Forelimbs 5, 9-10

247 T Pre-treatment No abnormalities detected

Treatment No abnormalities detected

Gestation No abnormalities detected

Lactation Staining Abnormal Colour, Brown, Head 10

248 T Pre-treatment No abnormalities detected

Treatment No abnormalities detected

Gestation No abnormalities detected

Lactation Coat Hair loss, Forelimbs 10

Hair loss, Hindlimb (Right) 5, 9

T Terminal sacrifice

Only animals with observations are presented
124

Huntingdon Life Sciences

WAG0015

APPENDIX 2 - continued Request ID: 284404

Clinical signs - individual observations for females - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Lactation Coat Hair loss, Forelimbs 1, 5, 9-10

Hair loss, Head 1, 5, 9

T Terminal sacrifice

Only animals with observations are presented
125

Huntingdon Life Sciences

WAG0015

APPENDIX 2 - continued Request ID: 284404

Clinical signs - individual observations for females - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Lactation Staining Abnormal Colour, Brown, Head 9-10

252 T Pre-treatment No abnormalities detected

Treatment No abnormalities detected

Gestation No abnormalities detected

Lactation Coat Hair loss, Forelimbs 10

253 T Pre-treatment No abnormalities detected

Treatment No abnormalities detected

Gestation No abnormalities detected

Lactation 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

Lactation Coat Hair loss, Forelimb (Left) 9-10

Hair loss, Forelimb (Right) 5, 9-10

Hair loss, Hindlimb (Right) 10

Hair loss, Ventral Body Surface 5, 9-10

T Terminal sacrifice

Only animals with observations are presented
126

Huntingdon Life Sciences

WAG0015

APPENDIX 2 - continued Request ID: 284404

Clinical signs - individual observations for females - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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-pregnant) 25

Comments: failed to litter 25

258 T Pre-treatment No abnormalities detected

Treatment No abnormalities detected

Gestation No abnormalities detected

Lactation Staining Abnormal Colour, Brown, Head 9-10

259 T Pre-treatment No abnormalities detected

Treatment No abnormalities detected

Gestation No abnormalities detected

Lactation 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

Lactation No abnormalities detected

T Terminal sacrifice

NP Not pregnant

Only animals with observations are presented
127

Huntingdon Life Sciences

WAG0015

APPENDIX 2 - continued Request ID: 284404

Clinical signs - individual observations for females - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Lactation Teeth Abnormal Colour, Pale 1, 5, 9-10

262 W Pre-treatment No abnormalities detected

Treatment No abnormalities detected

Gestation No abnormalities detected

Lactation 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

Lactation No abnormalities detected

265 TLL Pre-treatment No abnormalities detected

Treatment No abnormalities detected

Gestation No abnormalities detected

Lactation 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 presented
128

Huntingdon Life Sciences

WAG0015

APPENDIX 2 - continued Request ID: 284404

Clinical signs - individual observations for females - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Other Showing no interest in offspring Pre Day 1 lactation

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

Lactation 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 abnormalities detected

Treatment No abnormalities detected

Gestation Staining Abnormal Colour, Brown, Head 14, 20

Lactation 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 presented
129

Huntingdon Life Sciences

WAG0015

APPENDIX 2 - continued Request ID: 284404

Clinical signs - individual observations for females - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Lactation Staining Abnormal Colour, Brown, Head 5, 9-10

270 TLL Pre-treatment No abnormalities detected

Treatment No abnormalities detected

Gestation No abnormalities detected

Lactation No abnormalities detected

Reason for despatch Total litter loss 2

TLL Total litter loss 

T Terminal sacrifice

Only animals with observations are presented
130

Huntingdon Life Sciences

WAG0015

APPENDIX 2 - continued Request ID: 284404

Clinical signs - individual observations for females - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 Conformation Thin 12, 19

Coat Piloerection 8-10

Posture Hunched 8-10

Staining Abnormal Colour, Brown, Muzzle 11-12, 19

Gestation Behaviour Underactive 13-14

Build Conformation Thin 0

Coat Piloerection 13-15, 20, 25

Eyelids Partially Closed 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 abnormalities 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, Red, 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 presented
131

Huntingdon Life Sciences

WAG0015

APPENDIX 2 - continued Request ID: 284404

Clinical signs - individual observations for females - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 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 Piloerection 8-10

Posture Hunched 8-10

Gestation Teeth Abnormal Colour, Pale 20

Lactation Reason for Dispatch General poor clinical condition 2

Teeth Abnormal Colour, Pale 1

Reason for despatch Total litter loss 2

274 TR T Pre-treatment 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

Staining 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 presented
132

Huntingdon Life Sciences

WAG0015

APPENDIX 2 - continued Request ID: 284404

Clinical signs - individual observations for females - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 Underactive 24

Body temperature 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-treatment 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

Reason 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 presented
133

Huntingdon Life Sciences

WAG0015

APPENDIX 2 - continued Request ID: 284404

Clinical signs - individual observations for females - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Group 

/Sex

Animal 

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 condition 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 Failed To Litter 25

W Killed for welfare reasons

T Terminal sacrifice

NP Not pregnant

Only animals with observations are presented
134

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Eyelids (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 2
135

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 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 2
136

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 215 Behaviour

 Underactive 1 1, 4

 Unresponsive 1 1

Coat

 Piloerection 1 1-2, 4

Eyelids (Left)

 Partially Closed 2

Eyelids (Right)

 Partially 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 2
137

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 2
138

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 2
139

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 221 Behaviour

 Underactive 1-7 1-7

 Unresponsive 1, 3-4 1, 3-4

Breathing

 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 1
140

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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 1
141

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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 1
142

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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)

 Lachrymation 1

Eye (Right)

 Lachrymation 1
143

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 225 Behaviour

 Underactive 1-7 1-7

 Unresponsive 1, 3-4 1, 3-4

Breathing

 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 1
144

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 226 Behaviour

 Underactive 1-7 1-7

 Unresponsive 1, 3-4 1, 3-4

Breathing

 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 1
145

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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 1
146

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 228 Behaviour

 Underactive 1-7 1-7

 Unresponsive 1, 3-4 1, 3-4

Breathing

 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 1
147

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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 1
148

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 230 Behaviour

 Underactive 1-7 1-7

 Unresponsive 1, 3-4 1, 3-4

Breathing

 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 1
149

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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
150

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 232 Behaviour

 Underactive 1-7 1-7

 Unresponsive 1-7 1-7

Breathing

 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
151

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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
152

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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
153

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 235 Behaviour

 Underactive 1-7 1-7 1

 Unresponsive 1-7 1-7

Breathing

 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
154

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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
155

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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
156

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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
157

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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
158

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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
159

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 Closed 2
160

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 2
161

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

 Unresponsive 1 1

Coat

 Piloerection 1 1-2

Eyelids (Left)

 Partially Closed 2

Eyelids (Right)

 Partially Closed 2
162

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 2
163

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 Closed 2

Eyelids (Right)

 Partially Closed 2
164

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 261 Behaviour

 Underactive 1-3 1-3

 Unresponsive 1 1, 3

Breathing

 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 1
165

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 262 Behaviour

 Underactive 1-3 1-3

 Unresponsive 1 1, 3

Breathing

 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 1
166

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 263 Behaviour

 Underactive 1-3 1-3

 Unresponsive 1 1, 3

Breathing

 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 1
167

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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 1
168

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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 1
169

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 266 Behaviour

 Underactive 1-3 1-3

 Unresponsive 1, 3 1, 3

Breathing

 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 1
170

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 267 Behaviour

 Underactive 1-3 1-3

 Unresponsive 1 1, 3

Breathing

 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 1
171

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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 1
172

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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 1
173

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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 1
174

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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

Posture

 Hunched 2 2 2 2

Staining

 Other Colour (see comment) 2

Comments:

 brown staining head 2
175

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 272 FTM Behaviour

 Underactive 1-7 1-7

 Unresponsive 1-7 1-7

Breathing

 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 mate
176

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 Shallow 1 1-2

Coat

 Piloerection 1-2 1-2

Eyelids (Left)

 Partially Closed 1-2 1-2

Eyelids (Right)

 Partially Closed 1-2 1-2

Posture

 Hunched 2 2 2 2 2

Staining

 Other Colour (see comment) 2 2

Comments:

 brown staining head 2 2
177

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 274 Behaviour

 Underactive 1-2 1-2

 Unresponsive 1-2 1-2

Breathing

 Shallow 1 1-2

Coat

 Piloerection 1-2 1-2

Eyelids (Left)

 Partially Closed 1-2 1-2

Eyelids (Right)

 Partially Closed 1-2 1-2
178

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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

 Hunched 2

Staining

 Other Colour (see comment) 2 2 2 2

Comments:

 Brown staining head 2 2 2 2

FTM Failed to mate
179

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

 Underactive 1-3 1-3

 Unresponsive 1-3 1-3

Breathing

 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

Breathing

 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 mate
180

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 278 Behaviour

 Underactive 1-3 1-3

 Unresponsive 1-3 1-3

Breathing

 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

Breathing

 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 1-4
181

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 280 Behaviour

 Underactive 1-3 1-3

 Unresponsive 1-3 1-3

Breathing

 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
182

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 7
183

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 pregnant
184

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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

Breathing

 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, 19
185

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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

Breathing

 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, 19

U Unilateral implantation

TR Total resorption
186

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 265 Behaviour

 Underactive 0, 7, 14, 19 0, 7, 14, 19

Breathing

 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, 19 0, 7, 14, 19

 Unresponsive 19

Breathing

 Slow 0 0, 14, 19

Coat

 Piloerection 7 7

Eyelids (Left)

 Partially Closed 0 0, 14, 19

Eyelids (Right)

 Partially Closed 0 0, 14, 19
187

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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, 14, 19 7, 14, 19

 Unresponsive 7 7

Breathing

 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

Breathing

 Slow 7, 19

Coat

 Piloerection 14 14

Eyelids (Left)

 Partially Closed 19

Eyelids (Right)

 Partially Closed 19
188

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

 Underactive 0, 7, 19 0, 7, 14, 19

 Unresponsive 0

Breathing

 Slow 19 0, 7, 14, 19

Coat

 Piloerection 19 19

Eyelids (Left)

 Partially Closed 0, 7, 14

Eyelids (Right)

 Partially Closed 0, 7, 14

270 Behaviour

 Underactive 7, 14, 19 7, 14, 19

 Unresponsive 7 7

Breathing

 Slow 7, 19

Coat

 Piloerection 14 14

Eyelids (Left)

 Partially Closed 19

Eyelids (Right)

 Partially Closed 19
189

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 Slow 14, 19 7, 14, 19

Coat

 Piloerection 14 0, 7, 19 0, 7, 19

Eyelids (Left)

 Partially 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 resorption
190

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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

 Hunched 14 14

TR Total resorption
191

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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

Breathing

 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 resorption
192

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

Breathing

 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)

 Partially Closed 0, 7, 14, 19 0, 7, 14, 19

280 NP Behaviour

 Underactive 7, 14, 19 7, 14, 19

 Unresponsive 7, 14 7, 14

Breathing

 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

NP Not pregnant
193

Huntingdon Life Sciences

WAG0015

APPENDIX 3 - continued Request ID: 284713

Signs associated with dosing - individual observations for females during lactation - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 5
194

Huntingdon Life Sciences

WAG0015

APPENDIX 4 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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

2F 251 T Treatment Behaviour

 Underactive 1-7

 Unresponsive 1, 3

Coat

 Piloerection 2-7, 11, 14

Eyelids (Left)

 Partially Closed 14

Eyelids (Right)

 Partially Closed 14

Gestation Behaviour

 Underactive 7

Coat

 Piloerection 7, 19

Lactation No abnormalities detected

T Terminal sacrifice
195

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

2F 252 T Treatment Behaviour

 Underactive 1-7

 Unresponsive 1, 3

Coat

 Piloerection 2-7, 11, 14

Eyelids (Left)

 Partially Closed 14

Eyelids (Right)

 Partially Closed 14

Gestation Behaviour

 Underactive 0, 7, 14

Coat

 Piloerection 0, 7, 19

Lactation No abnormalities detected

T Terminal sacrifice 
196

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

2F 253 T Treatment Behaviour

 Underactive 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

Lactation No abnormalities detected

T Terminal sacrifice 
197

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

2F 254 T Treatment Behaviour

 Underactive 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

Lactation No abnormalities detected

T Terminal sacrifice 
198

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

2F 255 T Treatment Behaviour

 Underactive 1-7, 25

 Unresponsive 1, 3

Coat

 Piloerection 2-7, 11, 14, 25

Eyelids (Left)

 Partially Closed 14

Eyelids (Right)

 Partially Closed 14

Gestation Behaviour

 Underactive 0, 14

Coat

 Piloerection 0, 14

Lactation No abnormalities detected

T Terminal sacrifice 
199

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

2F 256 T Treatment Behaviour

 Underactive 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

Lactation No abnormalities detected

T Terminal sacrifice 
200

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

2F 257 NP T Treatment Behaviour

 Underactive 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 pregnant
201

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

2F 258 T Treatment Behaviour

 Underactive 1-7

 Unresponsive 1, 3

Coat

 Piloerection 2-7, 11, 14

Eyelids (Left)

 Partially Closed 14

Eyelids (Right)

 Partially Closed 14

Gestation Behaviour

 Underactive 0, 7, 14

Coat

 Piloerection 0, 7, 19

Lactation No abnormalities detected

T Terminal sacrifice 
202

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

2F 259 T Treatment Behaviour

 Underactive 1-7

 Unresponsive 1, 3

Coat

 Piloerection 2-7, 11, 14

Eyelids (Left)

 Partially Closed 14

Eyelids (Right)

 Partially Closed 14

Gestation Behaviour

 Underactive 7

Coat

 Piloerection 7, 19

Lactation No abnormalities detected

T Terminal sacrifice 
203

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

2F 260 T Treatment Behaviour

 Underactive 1-7

 Unresponsive 1, 3

Coat

 Piloerection 2-7, 11, 14

Eyelids (Left)

 Partially Closed 14

Eyelids (Right)

 Partially Closed 14

Gestation Behaviour

 Underactive 7

Coat

 Piloerection 7, 19

Lactation No abnormalities detected

T Terminal sacrifice 
204

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 261 T Treatment Behaviour

 Underactive 1-7, 11, 14, 18

 Unresponsive 1-3, 6-7, 18

Breathing

 Shallow 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

 Underactive 7, 14, 19

 Unresponsive 7

Breathing

 Slow 7, 19

Coat

 Piloerection 14

Eyelids (Left)

 Partially Closed 19

T Terminal sacrifice 
205

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 261 T Gestation Eyelids (Right)

 Partially Closed 19

Lactation Behaviour

 Underactive 9

T Terminal sacrifice 
206

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 262 W Treatment Behaviour

 Underactive 1-7, 11, 14, 18

 Unresponsive 1-3, 6-7, 18

Breathing

 Shallow 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

 Underactive 0, 7, 14, 19

 Unresponsive 7

Breathing

 Slow 0, 14, 19

Coat

 Piloerection 7, 14, 19

Eyelids (Left)

 Partially Closed 0, 7, 14, 19

W Killed for welfare reasons 
207

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 262 W Gestation Eyelids (Right)

 Partially Closed 0, 7, 14, 19

W Killed for welfare reasons 
208

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 263 U TR T Treatment Behaviour

 Underactive 1-7, 11, 14, 18

 Unresponsive 1-3, 6-7, 18

Breathing

 Shallow 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

 Underactive 0, 7, 14, 19

 Unresponsive 0

Breathing

 Slow 0, 7, 14, 19

Coat

 Piloerection 19

Eyelids (Left)

 Partially Closed 0, 7, 14

U Unilateral implantation 

T Terminal sacrifice 

TR Total resorption
209

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 263 U TR T Gestation Eyelids (Right)

 Partially Closed 0, 7, 14

U Unilateral implantation 

T Terminal sacrifice 

TR Total resorption
210

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 264 T Treatment Behaviour

 Underactive 1-7, 11, 14, 18

 Unresponsive 1-3, 6-7, 18

Breathing

 Shallow 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

 Underactive 0, 7, 14, 19

 Unresponsive 7

Breathing

 Slow 0, 14, 19

Coat

 Piloerection 7, 14, 19

Eyelids (Left)

 Partially Closed 0, 7, 14, 19

T Terminal sacrifice 
211

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 264 T Gestation Eyelids (Right)

 Partially Closed 0, 7, 14, 19

Lactation Behaviour

 Underactive 9

T Terminal sacrifice 
212

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 265 TLL Treatment Behaviour

 Underactive 1-7, 11, 14

 Unresponsive 1-3, 6-7

Breathing

 Shallow 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

 Underactive 0, 7, 14, 19

Breathing

 Shallow 0

 Slow 7, 19

Coat

 Piloerection 0, 14, 19

Eyelids (Left)

 Partially Closed 7, 19

TLL Total litter loss 
213

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 265 TLL Gestation Eyelids (Right)

 Partially Closed 7, 19

TLL Total litter loss 
214

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 266 TLL Treatment Behaviour

 Underactive 1-7, 11, 14, 18, 21

 Unresponsive 1-3, 6-7, 18, 21

Breathing

 Shallow 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

 Underactive 0, 7, 14, 19

 Unresponsive 19

Breathing

 Slow 0, 14, 19

Coat

 Piloerection 7

Eyelids (Left)

 Partially Closed 0, 14, 19

TLL Total litter loss 
215

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 266 TLL Gestation Eyelids (Right)

 Partially Closed 0, 14, 19

TLL Total litter loss 
216

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 Treatment Behaviour

 Underactive 1-7, 11, 14, 18

 Unresponsive 1-3, 6-7, 18

Breathing

 Shallow 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

 Underactive 7, 14, 19

 Unresponsive 7

Breathing

 Slow 7, 19

Coat

 Piloerection 14

Eyelids (Left)

 Partially Closed 19

TLL Total litter loss 
217

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 loss 
218

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 268 T Treatment Behaviour

 Underactive 1-7, 11, 14, 18

 Unresponsive 1-3, 6-7, 18

Breathing

 Shallow 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

 Underactive 7, 14, 19

 Unresponsive 7

Breathing

 Slow 7, 19

Coat

 Piloerection 14

Eyelids (Left)

 Partially Closed 19

T Terminal sacrifice 
219

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 268 T Gestation Eyelids (Right)

 Partially Closed 19

Lactation Behaviour

 Underactive 5, 9

Coat

 Piloerection 5

T Terminal sacrifice 
220

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 269 T Treatment Behaviour

 Underactive 1-7, 11, 14, 18

 Unresponsive 1-3, 6-7, 18

Breathing

 Shallow 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

 Underactive 0, 7, 14, 19

 Unresponsive 0

Breathing

 Slow 0, 7, 14, 19

Coat

 Piloerection 19

Eyelids (Left)

 Partially Closed 0, 7, 14

T Terminal sacrifice 
221

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 269 T Gestation Eyelids (Right)

 Partially Closed 0, 7, 14

Lactation Behaviour

 Underactive 5, 9

Coat

 Piloerection 5

T Terminal sacrifice 
222

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 270 TLL Treatment Behaviour

 Underactive 1-7, 11, 14, 18

 Unresponsive 1-3, 6-7, 18

Breathing

 Shallow 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

 Underactive 7, 14, 19

 Unresponsive 7

Breathing

 Slow 7, 19

Coat

 Piloerection 14

Eyelids (Left)

 Partially Closed 19

TLL Total litter loss 
223

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 270 TLL Gestation Eyelids (Right)

 Partially Closed 19

TLL Total litter loss 
224

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

4F 271 TR T Treatment Behaviour

 Underactive 1-8, 11, 14, 18, 21

 Unresponsive 1-2, 4-8, 11, 14, 18, 21

Breathing

 Shallow 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

 Underactive 0, 7, 14, 19

 Unresponsive 0, 19

Breathing

 Slow 7, 14, 19

Coat

 Piloerection 0, 7, 14, 19

T Terminal sacrifice

TR Total resorption 
225

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

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

 Underactive 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

Breathing

 Shallow 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 mate
226

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

4F 273 TLL Treatment Behaviour

 Underactive 1-8, 11, 14

 Unresponsive 1-2, 4-8, 11, 14

Breathing

 Shallow 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

 Underactive 0, 7, 14, 19

 Unresponsive 0, 14

Breathing

 Slow 0, 7, 19

Coat

 Piloerection 0, 7, 14

TLL Total litter loss
227

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

4F 273 TLL Gestation Eyelids (Left)

 Partially Closed 0, 7, 14, 19

Eyelids (Right)

 Partially Closed 0, 7, 14, 19

TLL Total litter loss 
228

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

4F 274 TR T Treatment Behaviour

 Underactive 1-7, 11, 14

 Unresponsive 1-2, 4-7, 11, 14

Breathing

 Shallow 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

 Underactive 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 resorption 
229

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

4F 275 FTM T Treatment Behaviour

 Underactive 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

Breathing

 Shallow 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:

 Brown staining head 8

T Terminal sacrifice

FTM Failed to mate
230

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

4F 276 W Treatment Behaviour

 Underactive 1-7, 11, 14, 18, 21

 Unresponsive 1-2, 4-7, 11, 14, 18, 21

Breathing

 Shallow 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

 Underactive 0, 7, 14, 19

 Unresponsive 0

Breathing

 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 reasons
231

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

4F 277 FTM T Treatment Behaviour

 Underactive 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

Breathing

 Shallow 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

FTM Failed to mate
232

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

4F 278 TR T Treatment Behaviour

 Underactive 1-7, 11, 14, 18

 Unresponsive 1-2, 4-7, 11, 14, 18

Breathing

 Shallow 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

 Underactive 7, 14, 19

 Unresponsive 7, 14

Breathing

 Slow 7, 19

Coat

 Piloerection 14

Eyelids (Left)

 Partially Closed 7, 19

Eyelids (Right)

 Partially Closed 7, 19

T Terminal sacrifice

TR Total resorption 
233

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

4F 279 W Treatment Behaviour

 Underactive 1-7, 11, 14, 18, 21, 25

 Unresponsive 1-2, 4-7, 11, 14, 18, 21, 25

Breathing

 Shallow 1-7, 14

 Slow 25

Coat

 Piloerection 1-7, 14, 18, 21

Eyelids (Left)

 Partially Closed 1, 3-7, 11, 14, 18, 21, 25

Eyelids (Right)

 Partially Closed 1, 3-7, 11, 14, 18, 21, 25

Gestation Behaviour

 Underactive 0, 7, 14, 19

 Unresponsive 0, 14, 19

Breathing

 Slow 0, 7, 14, 19

Coat

 Piloerection 14, 19

Eyelids (Left)

 Partially Closed 0, 7, 14, 19

Eyelids (Right)

 Partially Closed 0, 7, 14, 19

W Killed for welfare reasons
234

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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)

4F 280 NP T Treatment Behaviour

 Underactive 1-7, 11, 14, 18

 Unresponsive 1-2, 4-7, 11, 14, 18

Breathing

 Shallow 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

 Underactive 7, 14, 19

 Unresponsive 7, 14

Breathing

 Slow 7, 19

Coat

 Piloerection 14

Eyelids (Left)

 Partially Closed 7, 19

Eyelids (Right)

 Partially Closed 7, 19

T Terminal sacrifice

NP Not pregnant 
235

Huntingdon Life Sciences

WAG0015

APPENDIX 5 Request ID: 284396

Bodyweight - individual values for males - F0 generation (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

1M 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

210 269 319 374 381 401 421 450 472 475 479 501 510

2M 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

217 258 321 365 377 388 406 425 432 436 441 453 461

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

220 255 322 351 354 370 405 411 417 423 429 429 434
236

Huntingdon Life Sciences

WAG0015

APPENDIX 5- continued Request ID: 284396

Bodyweight - individual values for males - F0 generation (g)

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Day

/Sex Number 50 1M 201 534

202 479

203 512

204 563

205 570

206 591

207 529

208 545

209 570

210 524

2M 211 503

212 413

213 505

214 567

215 516

216 545

217 468

218 438

219 436

220 438
237

Huntingdon Life Sciences

WAG0015

APPENDIX 5 - continued Request ID: 284396

Bodyweight - individual values for males - F0 generation (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

3M 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

4M 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

240 252 322 341 353 368 385 406 408 416 418 416 418
238

Huntingdon Life Sciences

WAG0015

APPENDIX 5- continued Request ID: 284396

Bodyweight - individual values for males - F0 generation (g)

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

GroupAnimal Day

/Sex Number 50 3M 221 462

222 412

223 431

224 443

225 478

226 444

227 510

228 451

229 449

230 475

4M 231 468

232 406

233 380

234 410

235 457

236 459

237 403

238 385

239 447

240 423
239

Huntingdon Life Sciences

WAG0015

APPENDIX 6 Request ID: 284397

Bodyweight - individual values for females before pairing - F0 generation (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

1F 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

2F 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

260 209 235 253 256 264
240

Huntingdon Life Sciences

WAG0015

APPENDIX 6 - continued Request ID: 284397

Bodyweight - individual values for females before pairing - F0 generation (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

3F 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

269 195 215 220 225 232

270 192 217 231 232 247

4F 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

275FTM 205 230 238 241 237 236 235 248 257 247 249 251

276 202 223 222 232 237 216

277FTM 212 226 238 233 224 241 241 245 251 242 244 248

278 202 221 226 233 241

279 195 226 221 226 229 242

280 218 233 235 231 234

FTM Failed to mate
241

Huntingdon Life Sciences

WAG0015

APPENDIX 6 - continued Request ID: 284397

Bodyweight - individual values for females before pairing - F0 generation (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Group Animal Day Day

/Sex Number 50 53

4F 272FTM 252 255

275FTM 257 262

277FTM 242 243

FTM Failed to mate
242

Huntingdon Life Sciences

WAG0015

APPENDIX 7 Request ID: 284398

Bodyweight - individual values for females after mating - F0 generation (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

1F 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

2F 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 evaluation
243

Huntingdon Life Sciences

WAG0015

APPENDIX 7 - continued Request ID: 284398

Bodyweight - individual values for females after mating - F0 generation (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

3F 261 249 255 264 281 303 332 379

262 267 284 296 295 332 351 395

263U TR 266 284 286 297 308 282 287

264 246 268 286 297 326 347 401

265 237 259 274 285 303 322 355

266TLL 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

4F 271TR 244 251 271 281 265 275 282

273 227 234 239 248 272 281 316

274TR 238 258 264 281 293 307 320

276W 215 259 246 266 282 301 322

278TR 256 258 272 275 302 304 300

279W 251 262 270 280 294 289 289

280NP 235 261 260 264 265 257 264

U Unilateral 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, gestation data has been included from calculation of group means and statistical evaluation
244

Huntingdon Life Sciences

WAG0015

APPENDIX 8 Request ID: 284399

Bodyweight - individual values for females during lactation - F0 generation (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Group Animal Day Day Day

/Sex Number 1 5 10

1F 241 296 304 344

242 292 295 304

243 281 288 328

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

2F 251 302 321 328

252 279 290 284

253 281 285 306

254 340 356 377

255 319 321 344

256 296 321 327

258 285 282 302

259 290 306 309

260 309 317 322
245

Huntingdon Life Sciences

WAG0015

APPENDIX 8 - continued Request ID: 284399

Bodyweight - individual values for females during lactation - F0 generation (g)

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Group Animal Day Day Day

/Sex Number 1 5 10

3F 261 284 298 312

262W 310

264 310 309 305

265TLL 288

267TLL 304

268 290 294 293

269 284 296 277

270TLL 297

4F 273TLL 260

W Killed for welfare reasons, excluded from calculation of group means and statistical evaluation

TLL Total litter loss, excluded from calculation of group means and statistical evaluation
246

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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). 
247

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 90 101
248

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 evaluation
249

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 18 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

Unilateral implantation

Total resorption, excluded from calculation of group means and statistical evaluation

Killed for welfare reasons, excluded 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 included from calculation of group means and statistical evaluation
250

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 44

252 27 31

253 31 42

254 36 45

255 28 41

256 39 42

258 22 27

259 29 35

260 32 40
251

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 33
252

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 43 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 48 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 34
253

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 35
254

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

/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

54 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 mate
255

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 mate
256

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

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 leaked
257

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 75 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 evaluation
258

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 Unilateral 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 leaked
259

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 97 102 107

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 Unilateral 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
260

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 86 114
261

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 evaluation
262

Huntingdon Life Sciences

WAG0015

APPENDIX 15

Oestrous cycle length, mating performance, fertility and gestation length - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 period
263

Huntingdon Life Sciences

WAG0015

APPENDIX 15 - continued

Oestrous cycle length, mating performance, fertility and gestation length - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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)

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 sperm
264

Huntingdon Life Sciences

WAG0015

APPENDIX 15 - continued

Oestrous cycle length, mating performance, fertility and gestation length - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 5 229 + 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 – pregnancy indicated by implantation sites
265

Huntingdon Life Sciences

WAG0015

APPENDIX 15 - continued

Oestrous cycle length, mating performance, fertility and gestation length - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 - pregnancy indicated by implantation sites

2 Continuous few sperm NLL* No live litter - pregnancy indicated by late resorptions

3 Many scattered sperm

4 Solid masses of sperm

^ Estimated minimum cycle length - cycle in progress at start or end of smearing period

e Includes a period of extended oestrus (at least four consecutive days)
266

Huntingdon Life Sciences

WAG0015

APPENDIX 16

Litter size- individual values - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Implantations Total litter size Live litter size on Day

number Day1

15 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 1212

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 16
267

Huntingdon Life Sciences

WAG0015

APPENDIX 16 - continued

Litter size- individual values - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Implantations Total litter size Live litter size on Day

number Day1

15 10

2 251 14 14 14 14 14

252 13 12 1199

253 12 12 12 11 11

254 16 14 14 14 14

255 19 14 1377

256 14 14 13 12 12

257 NP0

25885555

259 149999

260 17 16 15 14 14

NP Not pregnant
268

Huntingdon Life Sciences

WAG0015

APPENDIX 16 - continued

Litter size- individual values - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Implantations Total litter size Live litter size on Day

number Day1

15 10

3 261 16 13 1298

262W 108

263 TR3

264 17 11 1166

265 TLL 125

266 TLL 114

267 TLL 162

268 16 11888

269 13 12 12 11 11

270TLL 16 14 14

W Killed for welfare reasons

TR Total resorption

TLL Total litter loss
269

Huntingdon Life Sciences

WAG0015

APPENDIX 16 - continued

Litter size- individual values - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Implantations Total litter size Live litter size on Day

number Day1

15 10

4 271 TR6

272 FTM0

273 TLL 1231

274 TR9

275 FTM0

276W 13

277 FTM0

278TR8

279W9

280 NP0

TR Total resorption 

FTM Failed to mate

TLL Total litter loss W Killed for welfare reasons

NP Not pregnant
270

Huntingdon Life Sciences

WAG0015

APPENDIX 17

Offspring survival indices - individual values - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 94 100 100
271

Huntingdon Life Sciences

WAG0015

APPENDIX 17 - continued

Offspring survival indices - individual values - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Post implantation Live birth Viability

number 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 pregnant
272

Huntingdon Life Sciences

WAG0015

APPENDIX 17 - continued

Offspring survival indices - individual values - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Post implantation Live birth Viability

number survival index index index

(%) (%) (%)

3 261 81 92 67

262W 800

263 TR0

264 65 100 55

265 TLL 420

266 TLL 360

267 TLL 130

268 69 73 100

269 92 100 92

270 TLL 88 1000

W Killed for welfare reasons

TR Total resorption

TLL Total litter loss
273

Huntingdon Life Sciences

WAG0015

APPENDIX 17 - continued

Offspring survival indices - individual values - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 TR0

272 FTM

273 TLL 25 330

274 TR0

275 FTM

276W0

277 FTM

278 TR0

279W0

280 NP

TR Total resorption

FTM Failed to mate

TLL Total litter loss W Killed for welfare reasons

NP Not pregnant
274

Huntingdon Life Sciences

WAG0015

APPENDIX 18

Sex ratio - individual values - F1 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 37.5
275

Huntingdon Life Sciences

WAG0015

APPENDIX 18 - continued

Sex ratio - individual values - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Total on Day Live on Day

number115 10

MF %MMF %MMF %MMF %M

2 25168 42.968 42.968 42.968 42.9

25248 33.338 27.336 33.336 33.3

25384 66.784 66.774 63.674 63.6

25468 42.968 42.968 42.968 42.9

25586 57.185 61.534 42.934 42.9

25676 53.876 53.866 50.066 50.0

257 NP

25823 40.023 40.023 40.023 40.0

25954 55.654 55.654 55.654 55.6

260 115 68.8 105 66.7 104 71.4 104 71.4

NP Not pregnant
276

Huntingdon Life Sciences

WAG0015

APPENDIX 18 - continued

Sex ratio - individual values - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Total on Day Live on Day

number115 10

MF %MMF %MMF %MMF %M

3 26148 33.348 33.336 33.335 37.5

262W53 62.5

263 TR

26474 63.674 63.624 33.324 33.3

265 TLL32 60.0

266 TLL31 75.0

267 TLL10 100.0

26844 50.044 50.044 50.044 50.0

26984 66.784 66.774 63.674 63.6

270 TLL68 42.968 42.9

W Killed for welfare reasons

TR Total resorption

TLL Total litter loss
277

Huntingdon Life Sciences

WAG0015

APPENDIX 18 - continued

Sex ratio - individual values - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Total on Day Live on Day

number115 10

MF %MMF %MMF %MMF %M

4 271 TR

272 FTM

273 TLL01 0.001 0.0

274 TR

275 FTM

276W

277 FTM

278 TR

279W

280 NP

TR Total resorption

FTM Failed to mate

TLL Total litter loss W Killed for welfare reasons

NP Not pregnant
278

Huntingdon Life Sciences

WAG0015

APPENDIX 19

Bodyweight - individual litter mean values for male offspring - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Day of age

number15 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

243 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 1.0
279

Huntingdon Life Sciences

WAG0015

APPENDIX 19 - continued

Bodyweight - individual litter mean values for male offspring - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Day of age

number15 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 pregnant
280

Huntingdon Life Sciences

WAG0015

APPENDIX 19 - continued

Bodyweight - individual litter mean values for male offspring - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Day of age 

number15 10

Mean SD Mean SD Mean SD

3 261 7.3 0.4 11.3 0.4 17.2 0.5

262W

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 loss
281

Huntingdon Life Sciences

WAG0015

APPENDIX 19 - continued

Bodyweight - individual litter mean values for female offspring - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Day of age 

number15 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 0.8 243 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 1.7
282

Huntingdon Life Sciences

WAG0015

APPENDIX 19 - continued

Bodyweight - individual litter mean values for female offspring - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Day of age 

number15 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 pregnant
283

Huntingdon Life Sciences

WAG0015

APPENDIX 19 - continued

Bodyweight - individual litter mean values for female offspring - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Day of age 

number1510

Mean SD Mean SD Mean SD

3 261 6.3 0.7 10.0 1.6 16.1 2.2

262W

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 loss
284

Huntingdon Life Sciences

WAG0015

APPENDIX 19 - continued

Bodyweight - individual litter mean values for female offspring - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Animal Day of age

number1

Mean SD

4 271 TR

272 FTM

273 TLL 5.5-

274 TR

275 FTM

276W

277 FTM

278 TR

279W

280 NP

TR Total resorption

FTM Failed to mate

TLL Total litter loss W Killed for welfare reasons

NP Not pregnant
285

Huntingdon Life Sciences

WAG0015

APPENDIX 20

Clinical signs - individual observations for offspring - F1 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 1F 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 presented
286

Huntingdon Life Sciences

WAG0015

APPENDIX 20 - continued

Clinical signs - individual observations for offspring - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

Group Dam

number

Number/Sex

of offspring

Observation Day of age

2 256 1F Bruising on head1

260 1F Pale1

Cold to touch1

Pup missing

3

3 2613M & 5F Dark in colour1

Underactive1

Cold to touch1-2

Slightly cold to touch3

1F Dark in colour1

Underactive1

Cold to touch1

Pup missing

2

1M & 1F Dark in colour1

Underactive1

Cold to touch1-2

Pups missing

3

1F Dark in colour1

Underactive1

Cold to touch1-2

Slightly cold to touch3

Pup missing

9

Only animals with observations are presented
287

Huntingdon Life Sciences

WAG0015

APPENDIX 20 - continued

Clinical signs - individual observations for offspring - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 touch1

No milk present in stomach1

Dark in colour1

Underactive1

Dam and pups killed for welfare reasons 

1

264 2M & 4F Cold to touch2

2M Cold to touch2

Pups missing

3

266 TLL 1M Cold to touch1

Dark in colour1

Inactive1

Killed for welfare reasons

1

267 TLL 1M No food present in stomach1

Dark in colour1

Underactive1

Killed for welfare reasons

1

Only animals with observations are presented W Killed for welfare reasons

TLL Total litter loss
288

Huntingdon Life Sciences

WAG0015

APPENDIX 20 - continued

Clinical signs - individual observations for offspring - F1 generation

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 touch1

Milk present in stomach1

1M Cold to touch1

Milk present in stomach1

Found dead

2

270 TLL 3M & 3F Cold to touch1

Pups missing

2

1MCold to touch1

Pale1

Pup missing

2

1M & 2F Cold to touch1

Found dead

2

1M & 3F Cold to touch1

Dark, underactive, little milk present in stomach2

Killed for welfare reasons

2

Only animals with observations are presented

TLL Total litter loss 
289

Huntingdon Life Sciences

WAG0015

APPENDIX 21

Macropathology - individual findings for offspring killed or dying before scheduled termination - F1 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

GroupF0 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 Milk 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 stomach
290

Huntingdon Life Sciences

WAG0015

APPENDIX 21 - continued

Macropathology - individual findings for offspring dying before scheduled termination (F1)

Group:1234

Compound: Control2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm):0 198 505 2900

GroupF0 Female 

Number

Death 

Code

Day of 

age

Sex of 

offspring

Macroscopic observation

3 261F <1F No milk in stomach

F1F Milk in stomach

262W <1M No milk in stomach

W <1F 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

265F <1 3 x M: 2 x F No milk in stomach

266W <1M No milk in stomach

F <1 2 x M No milk in stomach F <1 F Milk in stomach

267W <1M No milk in stomach

269F2M Milk in stomach

270F2 1 x M: 2 x F No milk in stomach

W2 1 x M: 3 x F No milk in stomach

Only animals with abnormalities presented
291

Huntingdon Life Sciences

WAG0015

APPENDIX 22

Macropathology - individual findings for offspring killed at scheduled termination - F1 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

GroupF0 Female 

Number

Sex of 

offspring

Macroscopic observation

2 254 F Left forepaw, third digit, pale swelling underlying tattoo, 4 mm

Only animals with abnormalities presented
292

Huntingdon Life Sciences

WAG0015

APPENDIX 23

Sperm analysis - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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) (millions/g) (million) (g) (millions/g) (million)

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 0.282 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 error
293

Huntingdon Life Sciences

WAG0015

APPENDIX 23 - continued

Sperm analysis - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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) (millions/g) (million) (g) (millions/g) (million)

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 2.00 208 416
294

Huntingdon Life Sciences

WAG0015

APPENDIX 23 - continued

Sperm analysis - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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) (millions/g) (million) (g) (millions/g) (million)

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 values
295

Huntingdon Life Sciences

WAG0015

APPENDIX 23 - continued

Sperm analysis - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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) (millions/g) (million) (g) (millions/g) (million)

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 1.95 168 328
296

Huntingdon Life Sciences

WAG0015

APPENDIX 24

Sperm motion data - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 10
297

Huntingdon Life Sciences

WAG0015

APPENDIX 24 - continued

Sperm motion data - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 5
298

Huntingdon Life Sciences

WAG0015

APPENDIX 24 - continued

Sperm motion data - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 values
299

Huntingdon Life Sciences

WAG0015

APPENDIX 24 - continued

Sperm motion data - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 6
300

Huntingdon Life Sciences

WAG0015

APPENDIX 25

Sperm morphology - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Group Animal Number of Normal Total abnormal Decapitate Head abnormal 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 1.0
301

Huntingdon Life Sciences

WAG0015

APPENDIX 25 - continued

Sperm morphology - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Group Animal Number of Normal Total abnormal Decapitate Head abnormal 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 0.0
302

Huntingdon Life Sciences

WAG0015

APPENDIX 25 - continued

Sperm morphology - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Group Animal Number of Normal Total abnormal Decapitate Head abnormal 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 values
303

Huntingdon Life Sciences

WAG0015

APPENDIX 25 - continued

Sperm morphology - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Group Animal Number of Normal Total abnormal Decapitate Head abnormal 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 0.0
304

Huntingdon Life Sciences

WAG0015

APPENDIX 26 Request ID: 284410

Organ weights - individual absolute values (g) for males - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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

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 1.82

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

219 439 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 1.98
305

Huntingdon Life Sciences

WAG0015

APPENDIX 26 - continued Request ID: 284410

Organ weights - individual absolute values (g) for males - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 1.99
306

Huntingdon Life Sciences

WAG0015

APPENDIX 27 Request ID: 289382

Macropathology and histopathology - individual findings - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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:

 Prostate, Rt. Epididymis, Rt. Testis
307

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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:

 Prostate, Rt. Epididymis, Rt. Testis
308

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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: 51 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. Testis
309

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 no gross observations and no findings microscopically:

 Prostate, Rt. Epididymis, Rt. Testis
310

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 Death: 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:

 Prostate, Rt. Epididymis, Rt. Testis
311

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (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 OBSERVATIONS

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. Testis
312

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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:

 Prostate, Rt. Epididymis, Rt. Testis
313

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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

 Interstitial Inflammation, Slight

The following examined tissues had no gross observations and no findings microscopically:

 Rt. Epididymis, Rt. Testis
314

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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:

 Prostate, Rt. Epididymis, Rt. Testis
315

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 Subgroup: 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:

 Prostate, Rt. Epididymis, Rt. Testis
316

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 211 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: 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:

 Prostate
317

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Spleen Spleen

 Capsule thickened, Multiple, areas. Capsular/Subcapsular Inflammation, Slight

 Adhesions/Inflammation/Fibrosis, Minimal

The following examined tissues had no gross observations and no findings microscopically:

 Prostate
318

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 213 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: 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:

 -
319

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 observations and no findings microscopically:

 -
320

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 Subgroup: 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:

 Prostate
321

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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/Fibrosis, Slight

The following examined tissues had no gross observations and no findings microscopically:

 Prostate
322

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 and no findings microscopically:

 Prostate
323

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 microscopically:

 -
324

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 Bodyweight: 438.7 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Spleen Spleen

 Capsule thickened, areas. Capsular/Subcapsular Inflammation, Minimal

 Capsular Thickening, Minimal

The following examined tissues had no gross observations and no findings microscopically:

 Prostate
325

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 had no gross observations and no findings microscopically:

 Prostate
326

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 Death: 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:

 -
327

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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:

 -
328

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 223 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: 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:

 -
329

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 224 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

Teeth

 Incisor(s) pale, Lower

The following examined tissues had no gross observations and no findings microscopically:

 -
330

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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:

 -
331

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

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

PATHOLOGY 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:

 Prostate
332

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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:

 -
333

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

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

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Prostate Prostate

 Small No significant lesions

Spleen Spleen

 Capsule 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:

 -
334

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 Acinar 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:

 -
335

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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

 Prominent tubules, A few

Teeth

 Incisor(s) pale, Lower

The following examined tissues had no gross observations and no findings microscopically:

 Prostate
336

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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

MACROPATHOLOGY HISTOPATHOLOGY

Prostate Prostate

 Small No significant lesions

Skin

 Hair loss, cranial region and sides 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 observations and no findings microscopically:

 Rt. Epididymis, Rt. Testis
337

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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. Epididymis
338

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 HISTOPATHOLOGY

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 observations and no findings microscopically:

 Rt. Epididymis, Rt. Testis
339

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 Death: 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. Testis
340

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 235 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: 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. Testis
341

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 Bodyweight: 453.1 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Prostate Prostate

 Small Interstitial Inflammation, Minimal

Spleen Spleen

 Capsule thickened, a few areas. Capsular/Subcapsular Inflammation, Minimal

 Adhesions, Anterior pole, capsule to adipose tissue. Capsular Thickening, Slight

 Adhesions/Inflammation/Fibrosis, Minimal

Teeth

 Incisor(s) pale, Lower

The following examined tissues had no gross observations and no findings microscopically:

 Rt. Epididymis, Rt. Testis
342

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 237 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: 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. Testis
343

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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. Testis
344

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 Spleen

 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. Testis
345

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 sacrifice

Date of Death: 07-Sep-12 Day of Death: 51 Week of Death: 8 Subgroup: 1 Terminal Bodyweight: 422.6 g

PATHOLOGY OBSERVATIONS

MACROPATHOLOGY HISTOPATHOLOGY

Prostate Prostate

 Small No significant lesions

Spleen Spleen

 Capsule thickened, Multiple, areas. Capsular/Subcapsular Inflammation, Minimal

 Capsular Thickening, Minimal

 Adhesions/Inflammation/Fibrosis, Minimal

Teeth

 Incisor(s) pale, Lower

The following examined tissues had no gross observations and no findings microscopically:

 Rt. Epididymis, Rt. Testis
346

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 241 Sex: Female Dose Group: 1 Phase: Lactation 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 gross observations and no findings microscopically:

 Ovaries
347

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 242 Sex: Female Dose Group: 1 Phase: Lactation 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 MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED***

The following examined tissues had no gross observations and no findings microscopically:

 Ovaries
348

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 243 Sex: Female Dose Group: 1 Phase: Lactation 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:

 Ovaries
349

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 244 Sex: Female Dose Group: 1 Phase: Lactation 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:

 Ovaries
350

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 245 Sex: Female Dose Group: 1 Phase: Lactation 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:

 Ovaries
351

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 246 Sex: Female Dose Group: 1 Phase: Lactation Sacrifice Status: Final phase sacrifice

Date of Death: 07-Sep-12 Day of Death: 10 Week of Death: 2 Subgroup: 1 Terminal Bodyweight: 378.7 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:

 Ovaries
352

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 247 Sex: Female Dose Group: 1 Phase: Lactation Sacrifice Status: 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:

 Ovaries
353

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 248 Sex: Female Dose Group: 1 Phase: Lactation 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 observations and no findings microscopically:

 Ovaries
354

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 249 Sex: Female Dose Group: 1 Phase: Lactation 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 RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED***

The following examined tissues had no gross observations and no findings microscopically:

 Ovaries
355

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 250 Sex: Female Dose Group: 1 Phase: Lactation 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:

 Ovaries
356

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 251 Sex: Female Dose Group: 2 Phase: Lactation 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

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:

 Ovaries
357

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 252 Sex: Female Dose Group: 2 Phase: Lactation Sacrifice Status: Final phase sacrifice

Date of Death: 04-Sep-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:

 Ovaries
358

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 253 Sex: Female Dose Group: 2 Phase: Lactation 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:

 Ovaries
359

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 254 Sex: Female Dose Group: 2 Phase: Lactation 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:

 Ovaries
360

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 255 Sex: Female Dose Group: 2 Phase: Lactation 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:

 Ovaries
361

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 256 Sex: Female Dose Group: 2 Phase: Lactation 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 OBSERVATIONS

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:

 Ovaries
362

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal 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:

 -
363

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 258 Sex: Female Dose Group: 2 Phase: Lactation Sacrifice Status: Final phase sacrifice

Date of Death: 05-Sep-12 Day of 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:

 Ovaries
364

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 259 Sex: Female Dose Group: 2 Phase: Lactation 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 gross observations and no findings microscopically:

 Ovaries
365

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 260 Sex: Female Dose Group: 2 Phase: Lactation Sacrifice Status: Final phase sacrifice

Date of Death: 06-Sep-12 Day of Death: 10 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:

 Ovaries
366

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 261 Sex: Female Dose Group: 3 Phase: Lactation 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:

 Ovaries
367

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 262 Sex: Female Dose Group: 3 Phase: Lactation 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, Present

The following examined tissues had no gross observations and no findings microscopically:

 Ovaries
368

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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:

 Ovaries
369

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 264 Sex: Female Dose Group: 3 Phase: Lactation 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

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:

 Ovaries
370

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 265 Sex: Female Dose Group: 3 Phase: Lactation 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

***ANIMAL HAS NO MACROSCOPIC FINDINGS RECORDED*** ***ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED***

The following examined tissues had no gross observations and no findings microscopically:

 Ovaries
371

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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:

 Ovaries
372

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 267 Sex: Female Dose Group: 3 Phase: Lactation 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

PATHOLOGY 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:

 Ovaries
373

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 268 Sex: Female Dose Group: 3 Phase: Lactation 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

MACROPATHOLOGY HISTOPATHOLOGY

Teeth

 Incisor(s) pale, Lower

The following examined tissues had no gross observations and no findings microscopically:

 Ovaries
374

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 269 Sex: Female Dose Group: 3 Phase: Lactation 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:

 Ovaries
375

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 270 Sex: Female Dose Group: 3 Phase: Lactation 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:

 -
376

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure 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 OBSERVATIONS

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:

 Ovaries
377

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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:

 -
378

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 273 Sex: Female Dose Group: 4 Phase: Lactation 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

MACROPATHOLOGY HISTOPATHOLOGY

Teeth

 Incisor(s) pale, Lower

The following examined tissues had no gross observations and no findings microscopically:

 Ovaries
379

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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:

 Ovaries
380

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

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: 241.6 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:

 -
381

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 welfare 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:

 Ovaries
382

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-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 Size of Corpora Lutea, Slight

Spleen Spleen

 Capsule thickened Capsular/Subcapsular Inflammation, Slight

 Adhesions, capsule to adipose tissue. Capsular Thickening, Moderate

 Adhesions/Inflammation/Fibrosis, Slight

Teeth

 Incisor(s) pale, Lower

The following examined tissues had no gross observations and no findings microscopically:

 -
383

Huntingdon Life Sciences

WAG0015

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-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

Exposure level (ppm) : 0 198 505 2900

Animal Number: 278 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: 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:

 Ovaries
384

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Macropathology and histopathology - individual findings - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 and pancreas. Capsular Thickening, Minimal

 Adhesions/Inflammation/Fibrosis, Minimal

 Extramedullary Haemopoiesis, Slight

Teeth

 Incisor(s) pale, Lower

The following examined tissues had no gross observations and no findings microscopically:

 Ovaries
385

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Macropathology and histopathology - individual findings - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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:

 -
386

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APPENDIX 28

Primordial ovarian follicle counts - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 24.5 17.5 3.0 21.0 21.5 21.0 32.0

SD 7.78 2.12 2.12 1.41 18.38 7.78 9.90 25.46

N 2 2 2 2 2 2 2 2
387

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APPENDIX 28 - continued

Primordial ovarian follicle counts - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 11.31 2.83

N 2 2
388

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Primordial ovarian follicle counts - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 16.5 18.0

SD 2.83 2.12 3.54 17.68 16.26 6.36 4.95 0.00

N 2 2 2 2 2 2 2 2
389

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APPENDIX 28 - continued

Primordial ovarian follicle counts - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 2
390

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APPENDIX 28 - continued

Primordial ovarian follicle counts - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 15.5 19.0 25.5 8.0 7.0 12.0 16.0

SD 1.41 12.02 9.90 9.19 0.00 2.83 14.14 4.24

N 2 2 2 2 2 2 2 2
391

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APPENDIX 28 - continued

Primordial ovarian follicle counts - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 14.5

SD 8.49 6.36

N 2 2
392

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APPENDIX 28 - continued

Primordial ovarian follicle counts - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 13.5 8.0 25.5 19.0 16.5 8.0

SD 4.95 2.83 9.19 4.24 20.51 1.41 2.12 7.07

N 2 2 2 2 2 2 2 2
393

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APPENDIX 28 - continued

Primordial ovarian follicle counts - individual values - F0 generation

Group : 1 2 3 4

Compound : Control 2-bromo-3,3,3-

trifluoropropene

2-bromo-3,3,3-

trifluoropropene

2-bromo-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 23.5 23.0

SD 6.36 7.07

N 2 2
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Annex 1 Certificate of analysis
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Annex 2 Aerosol technology and inhalation analysis report
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Working for a better future

Aerosol Technology Report

HLS study number: WAG0015

Author A. Grasiewicz

Version ID: Final

Issue date: 06 March 2013
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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 ............................................................................................................. 401

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 .................................................. 414
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Signature Page

Aerosol Technology Report
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1. Pre-study Atmosphere Characterization

Inhalation exposure system characterized including determinations of:

Achieved vapour concentration

Spatial distribution around the chamber

2. Test Article Handling and Processing for Inhalation 

Generation

As supplied, 2-Bromo-3,3,3-trifluoro-1-propene was easily vapourised using warmed air 

through a sintered glass vapouriser.

3. Exposure System

A schematic of the exposure system is presented in Figure 1. Components of the exposure 

system:

Directed flow Whole body exposure chamber:

0.75 m

3 chamber of stainless steel and glass construction

Animal Restraint:

Wire mesh modular caging

Aerosol Generator:

Sintered glass vaporiser for Groups 2 to 4 all in a water bath set at 40°C

Syringe driver, polypropylene syringe for Group 2

Liquid reservoir and feed rate used to alter concentration Groups 3 and 4

Test groups - 30 L/min

Diluent Airflow:

From in-house compressed air system – breathing quality

Group 1 - 149 L/min

Group 2 to 4 - 119 L/min

Extract Airflow:

Drawn by a fan extract system

Drawn from 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 

airflows

Chamber pressure differential monitored by Magnahelic set at -1 to 

-5 mm H2O to ensure a slight negative pressure within the chamber ~

150 L/min.
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4. Administration

Animals exposed for 6 hours per day, 7 days a week

Exposure system operational parameters monitored continuously

Airflows documented initially and monitored during exposure

5. Atmosphere Analysis

5.1 Vapour Concentration

Vapour samples collected as follows:

Sample frequency – minimum of 3 samples/group/exposure 

Sample location – representative animal level

Sample position remained constant

Sample 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 for calibration

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 spectrophotometer were used to calculate analysed concentration (ppm) using a 2nd order fit subjected to least 

squares regression analysis.

Standards were shared with another concurrent and related study.
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5.3 Nominal Concentration

A mean daily nominal concentration was calculated for all test groups from the mass of test 

article used over the exposure period and the airflow through the exposure chamber. The 

ideal gas equation was used with the molecular weight of the test article and the measured 

chamber conditions to compute the volume 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 

Monitored using a liquid in glass thermometer.

5.5 Chamber Humidity 

Monitored using wet and dry liquid in glass thermometers, with humidity calculated using 

proprietary tables.

6. Data and Calculations

To minimize the cumulative errors from repeated rounding of numbers, data in this report has 

been calculated continuously using unrounded numbers and only rounded for printing. 

Consequently, 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 Tables 1 to 3 and 

summarized below:

Group Concentration (ppm)

Target Mean

2 200 198

3 500 505

4 3000 2900

All groups were close to target.

Additional samples were taken for confirmation of atypical results and after minor generation

rate adjustments.

All samples collected for Group 1 were negative for the presence of test article. This 

information is not tabulated but is retained in the study data files.
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7.2 Chamber Temperature

The chamber temperature was monitored continuously for all groups and recorded at 

30 minute intervals. Daily means are presented in Table 4 and were within expected 

parameters for this type of exposure system.

7.3 Chamber Humidity

The chamber humidity was derived from wet and dry bulb thermometers for all groups. 

These temperatures were recorded at 30 minute intervals and the subsequent humidity 

determined from propriatory tables. Daily means are presented in Table 5 and were within 

expected parameters for this type of exposure system.
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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 1861188119011881

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 1801180118211811

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

48 205 220 216 214
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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

54 205 220 225 217

55 203 207 210 207

56 175 193 201 190

Mean 198

sd 18.2

CV (%) 9.2

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 lactation phase
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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

Sample 

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 5101480150514981

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 61115331508148215341

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

48 540 500 487 509
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Table 2 Chamber atmosphere 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

49 743 511 481 571 577

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 lactation phase
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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 38613148 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 298322193227292307122744

26 2615 2745 3040 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 221322861228092315022758

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 2984
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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 vessel 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 phase
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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

23 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 24.7 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 24.9
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Table 4 Chamber 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)
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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 38.1 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 47.2
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Table 5 Chamber 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)
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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

N M

P

Q

O

Key

A Whole body chamber J Water bath

B Test material delivery tubing K Generation flowmeters

C Elutriator L Control valves

D Secondary generation trolley M Balance for more accurate weighing

E Magnahelics N Harvard Syringe driver

F Pressure Vessels O Sampling port

G Load cells P Extract plenum inside chamber

H Air-line coil Q Drain for washing down

I Sintered glass vaporiser Closed during exposure
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Annex 3 Pathology report
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Working for a better future

PATHOLOGY REPORT 

2-bromo-3,3,3-trifluoropropene: Reproductive/Developmental 

Toxicity Screening Study in the CD Rat by Inhalation 

Administration 

HLS study number: WAG0015 

Version ID: Final 

Issue date: 13 November 2012 

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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 ...................................................................................................................... 9

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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 female exposed 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 kidney and 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. 

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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 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 examined 10 10 10 10 

 

Spleen 

Capsular thickening was seen in the 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 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 

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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 consistent with the common background 

of macroscopic changes seen at 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 was 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 treatment were 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 females exposed to 505 or 2900 ppm. 

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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/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 to mate (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 performed as expected up to the end of 

the study on Day 10 of lactation. 

The examination of the ovaries from the F0 generation females revealed the presence of 

histopathological changes attributable to treatment. A reduction in size of corpora lutea was 

observed in the ovaries of four females exposed to 2900 ppm and one female exposed to 

198 ppm. The corpora lutea were quite small and appeared to be not newly formed. As this 

change was not consistent with the increasing exposure level 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 for the irregular oestrus cycle or the prolonged gestation length 

as reported in the in life phase. 

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The histopathological examination of testes and epididymides from the F0 generation animals 

performed after scheduled treatment period revealed 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 within the 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 findings to correlate with the organ weight 

variation reported at necropsy. 

An increased incidence of small prostates was observed at the macroscopic examination. 

This was consistent with a reduction in prostate weight seen with increasing exposure level. 

No microscopic correlate for the small prostates could be identified. A low incidence of 

inflammatory changes was observed in all groups including 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 capsular/subcapsular inflammation 

and/or capsular thickening and/or adhesions/inflammation/fibrosis seen at the microscopic 

examination and are likely to be related to treatment. However, as no controls and only 

tissues with macroscopic changes were sampled and microscopically examined, conclusions 

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 

at the macroscopic examination could not be clarified as these lesions were not examined 

microscopically. 

Pale incisors and hair loss of the skin are not routinely presented for microscopic evaluation 

as such changes do not usually correlate with changes seen histopathologically. These tissues 

are also not routinely required to be examined for reproductive developmental screening 

studies. 

4. Conclusion 

The microscopic evaluation on this study revealed the presence of histopathological changes 

related to 2-bromo-3,3,3-trifluoropropene only in the ovaries of animals exposed to 

2900 ppm.

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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 Lutea 

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Annex 4 GLP compliance statements
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HUNTINGDON RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2011
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HUNTINGDON RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2012
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EYE RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2011
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EYE RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2012