Registration Dossier

Administrative data

Description of key information

In repeated dose studies, the principle effects of xylenes were adaptive changes in the liver, organ weight and body weight changes. Inhalation studies in rodents have demonstrated a potential to cause ototoxicity.
 

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11th April - 16th July 1986
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
no
Remarks:
There are no significant deviations which affected the quality or the integrity of the study or the interpretation of the results in the report.
Qualifier:
equivalent or similar to
Guideline:
EPA OPPTS 870.3100 (90-Day Oral Toxicity in Rodents)
Deviations:
no
Remarks:
There are no significant deviations which affected the quality or the integrity of the study or the interpretation of the results in the report.
Principles of method if other than guideline:
The aim of this study was to evaluate the subchronic toxicity of m-xylene when administered to rats daily by oral gavage for at least 90 days.
GLP compliance:
yes
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Test material name: m-xylene
- CAS number: 108-38-3
- State: clear colourless liquid
- Source: Fluka Chemical Corporation, Hauppauge, New York
- Sponsor: Dynamac Corporation
- Lot No.of test material: 259 211
- Expiration date of the lot/batch: not specified
- Purity: 99.1% (pure by GC-analysis)

- The test substance was analysed for identity and purity prior to study initiation and for purity after study termination.

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- During transit: stored frozen and protected from light
- Storage condition of formulated test material: room temperature

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: dosing preparations prepared using corn oil (vehicle) and were adjusted to 100% activity, based on a purity of 99%

Species:
rat
Strain:
Sprague-Dawley
Details on species / strain selection:
No further details specified
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories, Inc.
- Total number of animals: 168 males and 168 females
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 46 days
- Weight at study initiation: males = 203.7-246.4g and females = 142.1 - 174.8g
- Housing: rats were uniquely identified by ear tags and cage cards and were housed individually in elevated wire-mesh cages
- Diet (e.g. ad libitum): commercial rodent diet (Purina Certified Rodent Chow® #5002)
- Water (e.g. ad libitum): acidified tap water
- Acclimation period: acclimated to laboratory conditions for approximately 2 weeks

DETAILS OF FOOD AND WATER QUALITY: Contaminant analysis for the rodent chow 5002 diet and water can be found in the 'attached background material'

ENVIRONMENTAL CONDITIONS
- Temperature (°F): 70-77
- Humidity (%): 31-69
> temperature and relative humidity in the animal were recorded twice daily (with few exceptions)
- Photoperiod (hrs dark / hrs light): 12-hour light/dark cycle was maintained daily via artificial illumination

IN-LIFE DATES: From: 11th April To: 14-16th July 1986
Route of administration:
oral: gavage
Details on route of administration:
The formulated test material was given to the animals by oral gavage. The dosing volume was 2.5 mL/kg
Vehicle:
corn oil
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:
- Dosing solutions were prepared weekly.
- For each dose level, the amount of test material required was calculated and then weighed. It was transferred to a volumetric flask and then the vehicle, corn oil, was added to reach the final volume.
- The flask was capped and inverted by hand until thoroughly mixed.
- Groups and doses can be seen in Table 1 below.
Analytical verification of doses or concentrations:
yes
Remarks:
Formulated test material was analysed for purity, stability and dose concentration.
Details on analytical verification of doses or concentrations:
Stability results reveal that there was a small amount of test material lost after 21 days of storage at 5°C.
All samples analysed were within the specific limits ± 10% of target concentration
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Daily
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Group 1 (control) and Group 5 (baseline)
Dose / conc.:
100 mg/kg bw/day (nominal)
Remarks:
Group 2
Dose / conc.:
200 mg/kg bw/day (nominal)
Remarks:
Group 3
Dose / conc.:
800 mg/kg bw/day (nominal)
Remarks:
Group 4
No. of animals per sex per dose:
20 animals/sex/dose
Control animals:
yes, concurrent vehicle
Details on study design:
- Rationale for animal assignment : Sprague-Dawley rats were employed since they have historically been used in safety evaluation studies and were requested by the EPA
- Animal selection: following physical and ophthalmologic examinations, a total of 85 clinically acceptable rats of each sex were randomly allocated to 4 treatment groups using a computer weight randomisation program, each composed of 20 animals/sex/group and 5 animals/sex were assigned to the baseline group employed for pre-treatment bloodwork.
- Animals not selected were assigned to another study or discarded without necropsy.
- Baseline groups were used for pre-treatment bloodwork and later discarded without necropsy.
Positive control:
No positive control was employed.
Observations and examinations performed and frequency:
- All rats were observed daily for clear signs of toxicity, beginning about one hour after dosing
- A cursory examination was performed during dosing

PHYSICAL EXAMINATIONS: Yes
- Time schedule: performed at initiation and weekly thereafter

MORIBUNDITY/MORTALITY CHECKS: Yes
- Time schedule: twice daily

INDIVIDUAL BODY WEIGHTS: Yes
- Time schedule for examinations: recorded during the randomisation procedure, at initiation and weekly thereafter

FOOD CONSUMPTION : Yes
- Food consumption for each animal determined: Yes, recorded weekly

FOOD EFFICIENCY: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OPHTHALMOSCOPIC EXAMINATION: Yes
- Performed by a board-certified veterinary opthalmologist
- Time schedule for examinations: prior to treatment, Week 13
- Dose groups that were examined: all animals prior to treatment and the last 10 survivors per sex per group during Week 13
- Pupils of both eyes were dilated prior to examination using 1% Atropine Sulphate

HAEMATOLOGY: Yes
- Time schedule for collection of blood: prior to study initiation, weeks 5-13
- Anaesthetic used for blood collection: Yes - samples were collected via the orbital sinus while under carbon dioxide anaesthesia
- Animals fasted: Yes, overnight
- How many animals: 5 animals/sex (Baseline group) prior to initiation and on the first 10 survivors/sex/group during weeks 5-13
- Parameters examined: please see Table 2 below

CLINICAL PATHOLOGY: Yes
- Time schedule for collection of blood: prior to study initiation, weeks 5-13
- Animals fasted: Yes - samples were collected via the orbital sinus while under carbon dioxide anaesthesia
- How many animals: 5 animals/sex (Baseline group) prior to initiation and on the first 10 survivors/sex/group during weeks 5-13
- Parameters examined: please see Table 3 below

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
- For all animals that died during the study and all survivors that were sacrificed after 13 weeks of treatment and were given a complete gross necropsy. Please see Table 4 below.
- Euthanasia was exsanguination whilst under sodium pentoarbital anaesthesia.
- At the terminal sacrifice, five animals per sex from the control group and 10 animals per sex from each treated group were selected for electron microscopy, the results of which will be available in a separate report.

HISTOPATHOLOGY: Yes
- All preserved tissues from all animals were embedded in paraffin, sectioned, and stained with haemoxylin and eosin.
- The following tissues were examined microscopically:
(1) All tissues from control and high-dose animals and from all animals not surviving to termination.
(2) Gross lesions, lungs, liver and kidneys from all remaining animals.
Other examinations:
ORGAN WEIGHTS
-The following organs were weighed for all terminally killed animals, except those selected for electron microscopy:
liver, kidneys, spleen, adrenal glands, brain with stem, heart, tested with epididymides, ovaries

TISSUE PRESERVATION
- Please see Table 5 below for all tissues that were preserved from each animal (when present) in 10% neutral buffered formalin.
Statistics:
- Cumulative survival data were analysed using the National Centre Institute Package (Life Table Analysis).
- Trend analysis of survival was evaluated at the 5.0% one-tailed probability level.
- Body weights, food consumption, appropriate clinical pathology data and organ weight data were compared between the control and treated groups of the same sex
- Tests for homogeneity of variances and ANOVA were evaluated at the 5.0% two-tailed probability level.
- Control vs. treated group means were evaluated using Dunnett's t-test at the 5.0% two-tailed probability level.
- Data transformations were performed where variances were heterogenous, which are indicated in the tables with an appropriate superscript next to the control group mean. The list of these superscripts can be seen in Table 6 below.
- When an effect is referred to as 'significant', it means that there is a statistically significant difference between the control group and the specified treated group of the same sex
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
Urine stains were observed weekly in both sexes of Group 4 and thought to be possibly treatment-related.
Other common weekly signs observed were chromodacryorrhea and malocclusion, but these are not thought to be treatment-related.
Salivation was frequently observed in both sexes of Group 4 and only rarely in the other treated groups. It occurred just prior to dosing and the animals returned to normal by the one-hour post-dosing observations.
Other daily observations included hyperactivity, convulsions and epistaxis - they were only observed in the first three weeks of the study at a low frequency
Mortality:
mortality observed, non-treatment-related
Description (incidence):
Males: there was a significantly higher mortality in Group 3 (5 males died), but there was not a significant trend.

Females: there was significant trend and a significantly higher mortality in Group 3 (5 females died) and Group 4 (4 females died).

Males and females: In the 19 unscheduled deaths, all but one of the animals had foreign material in the alveoli and were determined to be related to gavage dosing accidents rather than being related to the test material (please refer to the histopathological findings section). This may also relate to the lung lesions, mottled lungs and failure of the lungs to collapse seen in males and females of Groups 3 and 4.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
The mean body weight gain over the course of the experiment was significantly decreased in Groups 3 and 4 for males and in Group 4 for females.
There was a clear relationship between dose and body weight reduction in both sexes.
The overall total mean body weights (Groups 1-4) were 15% and 8% lower than in the controls for males and females respectively.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Statistical analysis was employed at three intervals - Weeks 1-5, 6-9 and 10-13.
Food consumption was significantly reduced during Weeks 1-5 for Group 4 males and during Weeks 6-9 for Group 3 and 4 males.
Food consumption was comparable for all female groups at all three intervals.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Only two animals (one Group 1 male and one Group 3 male) showed abnormalities at the terminal examination and thus were judged to be unrelated to treatment.
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
The haemotocrit was significantly decreased in Group 4 females.
At Week 13, there were no significant changes to the haematology parameters in either sex.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Significant changes were observed at the Week 5 interval:
- Group 4 males - potassium was increased and total bilirubin was decreased.
- Group 4 females - cholesterol and calcium increased
- Group 3 and 4 females - potassium was increased.

There were fewer significant changes at Week 13:
- Group 4 males - alanine aminotransferase was increased
- Group 4 females - calcium and cholesterol was increased
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
Group 4:
- the terminal body weights were significantly decreased in both sexes
- males - there were few significant changes in organ weights, however heart weight was decreased and relative brain and kidney weights were increased vs. control
- females - no significant changes in absolute or relative organ weights vs. control
Gross pathological findings:
effects observed, non-treatment-related
Description (incidence and severity):
There were few gross lesions observed.
The most commonly seen gross lesions were mottled lungs and failure of the lungs to collapse.
These observations were most frequently observed in both sexes of Group 3 and 4, but were only found in the animals found dead during the experiment.
The other lesions observed were sporadic and showed no apparent dose relationship
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Several spontaneously-occurring incidental lesions were observed in animals at the terminal sacrifice, but were concluded to be not related to treatment.

In the 19 unscheduled deaths (males Group 2-4; females Group 3-4), all but one of the animals had foreign material in the alveoli and were determined to be related to gavage dosing accidents rather than related to the test material.
Histopathological findings: neoplastic:
not examined
Key result
Dose descriptor:
NOAEL
Remarks:
m-xylene
Effect level:
ca. 200 mg/kg bw/day (nominal)
Sex:
female
Basis for effect level:
body weight and weight gain
Remarks on result:
other: It has been reported as NOEL in the report
Key result
Dose descriptor:
NOAEL
Remarks:
m-xylene
Effect level:
ca. 100 mg/kg bw/day (nominal)
Sex:
male
Basis for effect level:
body weight and weight gain
Remarks on result:
other: It was reported as NOEL in the report.
Key result
Critical effects observed:
not specified

Table 7: Summary incidence of clinical signsa

 

Males

Females

Observation

Group 1

Group 2

Group 3

Group 4

Group 1

Group 2

Group 3

Group 4

Malocclusion

2

2

2

0

1

0

0

0

Salivation

0

0

0

0

0

0

0

1

Urine stains

0

0

0

3

0

0

0

4

Alopecia

0

0

0

1

0

0

2

1

Sores

0

1

0

2

2

0

0

0

Bloody crust

1

0

0

2

1

0

0

0

Chromodacryorrhea

3

1

2

2

2

0

0

0

Small moveable tissue mass

0

0

0

1

0

0

0

0

aTotal incidence for entire study

Table 8: Overall mean body weight change (Week 0-13)

Sex

Group

Dose level (mg/kg)

Body weight

Mean

S.D.

N

Male

1

0

293.3

43.46

20

2

100

289.5

30.61

17

3

200

265.2S

30.37

15

4

800

219.5S

26.16

18

Female

1

0

115.9

18.00

20

2

100

113.2

14.06

20

3

200

107.2

14.86

16

4

800

98.1S

16.21

16

Sstatistically significant result

Table 9: Mean total food consumption

Sex

Group

Dose level (mg/kg)

 

Week

1-5

6-9

10-13

Male

1

0

Mean

860.0A

667.8

644.3

S.D.

66.03

48.31

52.29

N

10

20

9

2

100

Mean

831.4

640.6

611.6

S.D.

46.07

42.88

39.33

N

6

14

6

3

200

Mean

849.5

661.7S

614.6

S.D.

25.81

30.73

23.15

N

6

14

4

4

800

Mean

773.6S

612.0S

586.8

S.D.

59.64

55.08

28.92

N

8

14

5

Female

1

0

Mean

634.9

511.4

472.4

S.D.

56.91

46.89

28.32

N

8

13

9

2

100

Mean

650.1

505.4

478.0

S.D.

56.50

48.52

52.80

N

9

17

8

3

200

Mean

678.9

493.4

465.6

S.D.

38.24

50.78

40.62

N

5

15

6

4

800

Mean

656.8

479.7

488.8

S.D.

46.35

33.23

43.20

N

5

10

6

Adata analysed following log10transformation

SStatistically significant result

Table 10: Mean body weight changes and the percentage difference from controls.

Doses (mg/kg bw/day)

0

100

200

800

Males (g)

293.2

289.5

265.2 (s)

219.5 (s)

% difference from controls

--

1.26

9.5

25.1

Females (g)

115.9

113.2

107.2

98.1 (s)

% difference from controls

--

2.33

7.5

15.4

S = significant result

Conclusions:
Based on the results of this study and following the assumption that the vehicle and/or test material aspiration were responsible for the deaths in the treated animals, the no-observed-adverse-effect level (NOAEL) of m-xylene was 100 mg/kg/day for males and 200 mg/kg/day for females.
Executive summary:

The sub-chronic toxicity of m-xylene was investigated in this study on male and female Sprague-Dawley rats. The rats were acclimated for approximately 2 weeks and were 46 days of age at the start of the study. The rats underwent physical and ophthalmologic examinations prior to the test and then 85 clinically acceptable rats of each sex were assigned to treatment groups, each composed of 20 rats/sex plus 5 rats/sex used in the baseline haematology group. The rats were gavaged daily at each sex were tested at each dose daily for at least 90 days.

 

Dosing solutions were prepared weekly. For each dose level, the calculated amount of test substance, m-xylene, was weighed and transferred to a volumetric flask. The vehicle, corn oil, was added to achieve the final volume and then the flask was capped and inverted by hand until thoroughly mixed. M-xylene was administered to the rats daily via oral gavage at the following dose levels; 0, 100, 200 and 800 mg/kg/day (Groups 1 -4 respectively). The control group (Group 1) received the vehicle only and the Baseline group (Group 5 0 mg/m^3) were used for pre-treatment bloodwork and later discarded without necropsy. Several observations were recorded, including clinical signs, body weights, food consumption, opthalmologic examinations, organ weights and organ-to-body weight ratios, mortality, gross pathology, clinical chemistry and histopathology was recorded.

 

Mortality was found to be significantly increased in male Group 3 and female Group 3 and 4, however based on histopathology findings, it is thought that the cause of these deaths was most likely to be due to vehicle and/or compound aspiration. In the 19 unscheduled deaths seen in males (Group 2 -4) and females (Groups 3 -4), all but one of the animals had foreign material in the alveoli and were determined to be related to gavage dosing accidents rather than being related to the test material, which may also relate to the lung lesions, mottled lungs and failure of the lungs to collapse seen in males and females of Groups 3 and 4, which were only observed in the animals found dead. Salivation was frequently observed in both sexes of Group 4 and only rarely seen in the other treatment groups. Urine stains were also observed weekly in both sexes of Group 4 and thought to be possibly-treatment related. Other commonly observed clinical signs were chromodacryorrhea and malocclusion, but were not thought to be treatment-related. Food consumption was significantly reduced for males during Weeks 1-5 for Group 4 males and during Weeks 6-9 for Group 3 and 4 males. The results remained comparable throughout the study for female rats. Only two animals showed abnormalities in the ophthalmological examination and thus it was concluded to be unrelated to the treatment. Few clinical pathology parameters were significantly changed and all were observed in Group 4, except for one seen in female Group 3. Few significant changes were seen in absolute or relative organ weights, all of which were seen in Group 4 males, where heart weight was decreased and relative brain and kidney weights were increased.

The mean body weight change was significantly lower than controls for males in Group 3 (200 mg/kg bw/day) and Group 4 (800 mg/kg bw/day) and for females in Group 4 (800 mg/kg bw/day) (Table 10). The percentage difference in mean body weight change between the control and each dose group for males and females were calculated. The percentage difference for males was 9.5% and 25.1% for Group 3 and 4 males respectively and 15.4% for Group 4 females. Reduced absolute body weight of approximately >10% is generally considered to be a sign of general toxicity, therefore the dose-related reductions in Group 3 and 4 males and Group 4 females are considered to be adverse and are therefore have been listed as NOAELs, whilst they have been reported as NOELs in the report.

Based on the results of this study and following the assumption that the vehicle and/or test material aspiration was responsible for the deaths in the treated animals in groups 3 and 4, the no-observed-effect level (NOAEL) of m-xylene was 100 mg/kg/day for males and 200 mg/kg/day for females.

Endpoint:
sub-chronic toxicity: oral
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
11 July 2019 to 23 January 2020
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Justification for type of information:
Read-across justification is included in the Section 13 reports "Read-Across Justification Document Xylene" and "Xylene Isomers Category Justification Document".
Reason / purpose:
read-across source
Reason / purpose:
read-across: supporting information
Qualifier:
according to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.3100 (90-Day Oral Toxicity in Rodents)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Wistar
Details on species / strain selection:
The rat is a suitable rodent species for toxicity testing, acceptable to regulatory authorities and for which extensive background data are available.
Sex:
male/female
Details on test animals and environmental conditions:
40 male and 40 female rats of the Crl:WI(Han) strain were supplied by Charles River (UK) Limited, Margate, Kent, CT9 4LT, England. The animals were 5 to 6 weeks of age on arrival and on examination were found to be healthy. After 12 days acclimatisation, they were re-examined and confirmed to be suitable for use. On the first day of dosing the males weighed 169 to 231 g and the females weighed 141 to 184 g.

Allocation to groups was performed using the Provantis stratified randomisation procedure based on individual body weights recorded on arrival. The cages were positioned in the battery using a specific cage-plan. Each animal was uniquely identified by a subcutaneously implanted micro-identification device. To aid identification, animals were tail marked with their own unique identification numbers using an indelible pen.

Animals were housed in groups up to five, by sex, in solid-floor cages with bedding provided. Bedding was changed as often as necessary to maintain hygiene.
The study room was illuminated to give a cycle of 12 hours light and 12 hours dark and was air-conditioned. The target ranges for temperature and humidity were 19 °C to 23 °C and 40 % to 70 %, respectively. Recorded values were within, or only marginally outside, these limits.

A pelleted diet, Teklad 2014C Rodent Maintenance Diet, supplied by Envigo RMS (UK) Limited, and mains tap water (in bottles) were freely available.
Certificates of analysis for diet and water are held centrally and retained within the Sequani archives. It was considered that none of the contaminants that were monitored was present at a level that might have prevented the study objective from being achieved.
Route of administration:
oral: gavage
Details on route of administration:
Animals were dosed once daily for at least 90 days, by gavage using a rubber catheter and disposable syringe, at a constant dose volume of 4 mL/kg body weight, until necropsy. Individual doses were adjusted according to the most recent body weight.
Vehicle:
corn oil
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Samples were sent to the Principal Investigator, Dale Burton, at Covance, Eye, Suffolk, IP23 7PX, England for analysis (Covance reference number: WP50YW). The method of analysis used to analyse the dose formulation was validated in Covance Study Number TK45GM
Duration of treatment / exposure:
at least 90 days
Frequency of treatment:
once daily
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Group 1
Dose / conc.:
100 mg/kg bw/day (nominal)
Remarks:
Group 2
Dose / conc.:
300 mg/kg bw/day (nominal)
Remarks:
Group 3
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
Group 4
No. of animals per sex per dose:
10 animals per sex per dose
Control animals:
yes, concurrent vehicle
Details on study design:
A total of 80 animals were used in this study, where animals were dosed once daily for at least 90 days, by gavage using a rubber cather and disposable syringe, at a constant dose volume of 4 mL/kg body weight, until necropsy. Individual doses were adjusted according to the most recent body weight.
Observations and examinations performed and frequency:
Clinical observations
- Animals were examined twice daily for mortality and mobility
- Detailed clinical examination once each week
- Animals were observed approx. 1 and 2 hours after dosing
- Following the onset of clinical signs, animals were observed at approximately 15 minutes and at one and around four hours after dosing and at the weekend they were observed either at one hour after dosing or at the end of the working day, whichever was sooner
Ophthalmoscopy
- Both eyes of all animals were examined before the start of treatment
Body weight
- All animals were weighed at the start of dosing and then weekly until necropsy
Food intake
- During the treatment period, the amount of food consumed by each cage of animals was recorded weekly until necropsy.
Behavioural observations
- All animals were observed once weekly, starting predose, for their behaviour both within their cage and then after placement in an open arena
Functional observation battery
- During Week 12/13, sensorimotor responses to visual, acoustic, tactile or proprioceptive stimuli, grip strength and motor activity were recorded for all animals on two occasions for each animal, one before dosing and the second after dosing on the same day
Sacrifice and pathology:
Sacrifice
At the end of the treatment period, all animals were killed by exposure to carbon dioxide gas in a rising concentration, weighed and examined externally. The abdominal cavity was opened and the animals were exsanguinated from the caudal vena cava. The cranial and thoracic cavities were opened, a full internal examination was performed and all macroscopic abnormalities were recorded and retained.

Pathology
- macroscopic and microscopic pathology on:
Macroscopic and microscopic pathology on adrenal glands, pancreas, aorta, pituitary, bone marrow smear, prostate & seminal vesicles (incl. coagulating gland), brain (10 levels examined), rectum, caecum, salivary gland (submandibular), colon, sciatic nerve (LS and TS), duodenum, site of mammary gland, epididymides, skeletal muscle, eyes (incl. optic nerves), skin, femur & join (incl. Marrow), spinal cord (3 levels examined LS and TS), Harderian glands, spleen, heart, stomach, ileum, (incl. Peyer’s patch), submandibular lymph nodes, jejunum, testes, kidneys, thymus, lacrimal glands, thyroids (incl. parathyroids), larynx, trachea, liver, urinary bladder, lungs (incl. mainstem bronchi), uterus (incl. uterine cervix and oviducts), mesenteric lymph nodes, vagina, oesophagus, all gross lesions, and ovaries.
Other examinations:
Clinical laboratory studies
- Blood sample collection: blood samples (1.5 mL) were taken under isoflurane anaesthesia from the sublingual vein of all animals on the day of necropsy (0.5 mL into EDTA for haematology; 0.5 mLinto 3.2% w/v aqueous trisodium citrate for coagulation; 0.5 mL into lithium heparin for blood chemistry).
1. Haematology:
Haemoglobin concentration (Hb), platelet count (plate), red blood cell count (RBC), total leucocyte count (WBC), packed cell volume (PCV), neutrophils (Neut), mean cell volume (MCV), lymphocytes (Lymph), mean cell haemoglobin (MCH), monocytes (Mono), mean cell haemoglobin concentration (MCHC), eosinophils (Eosin), mean platelet volume (MPV), basophils (Baso), mean corpuscular volume retics (MCHr), large unstained cells (LUC), platelet distribution width (PDW), reticulocytes (Retics), cellular haemoglobin retics (CHr), plateletcrit (PCT), red blood cell distribution width (RDW), absolute reticulocyte count (A Retic), haemoglobin distribution width (HDW), and cell morphology*)
(*blood smears were prepared and were examined when warranted by machine morphology flags. All parameters were measured on the ADVIA 120 haematology analyser)
2. Coagulation (measured on the ACL Elite Pro):
Prothrombin time (PT) and activated partial thromboplastin time (APTT)
3. Blood chemistry:
Urea, globulin (Glob), creatinine (Cluc), albumin/globulin ratio (A/G), glucose (Gluc), total bilirubin (BiliT), alkaline phosphatase (ALP), cholesterol (Chol), alanine aminotransferase (ALT), sodium (Na), aspartate aminotransferase (AST), chloride (Cl), bile acids (B.Acids), calcium (Ca), total protein (T.Prot), potassium (K), albumin (Alb), HDL cholesterol (HDL), and cholesterol (LDL).

Thyroid hormone assessment - blood sampling and analysis

Vaginal smears - taken by lavage, examined under light microscopy and the stage of oestrous cycle was determined by the type of cell present.but

Organ weights: the following organs were weighed after trimming of fat and other contiguous tissue (contralateral organs were weighed together):
Adrenal glands, prostate & seminal vesicls (incl. coagulating gland), brain, epididymides, spleen, heart, testes, kidneys, thymus, liver, thyroids (incl. parathyroids), ovaries, uterus (incl. uterine cervix), and pituitary
Statistics:
Data were processed to give group mean values and standard deviations, where appropriate. Where the data allowed, the following methods were used for statistical analysis, comparing Groups 2, 3 and 4 against Group 1.

Depending on the nature of the data set that was to be analysed, appropriate tests were applied. Where parametric tests were appropriate they were preceded by a check for homogeneity of variance using the Levene test and, where available, the Shapiro-Wilks test for normality. If either of these two assumptions failed, a log transformation was applied before retesting. If the transformation failed, appropriate non-parametric tests were applied.

Probability values of less than 5 % were regarded as providing sufficient evidence to reject the null hypothesis and therefore statistical significance was identified at the p<0.05 level. For illustrative purposes, significance levels of p<0.01 and p<0.001 were also noted.
Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Males and females given 1000 mg/kg/day showed decreased activity and unsteady gait from Days 19/18 of dosing onwards (from approximately 15 minutes after dosing persisting for up to four hours), respectively. The incidence of these clinical signs decreased with time and by Day 72 of dosing, only one male was showing decreased activity and abnormal gait and over Days 84 to 92 of dosing; at the most, eight out of ten females showed these signs after dosing.
Mortality:
no mortality observed
Description (incidence):
There were no deaths during the study
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
For the first 3 weeks of dosing the body weights of males given 300 or 1000 mg/kg/day were very close to those of the Controls but by Week 4 it appeared that they were starting to diverge from the Controls and by Week 5 this was more pronounced. The Week 5 body weights of males given 300 or 1000 mg/kg/day were 5 % lower than those of the Controls and their body weight gains over Weeks 1 to 5 were 13 % and 15 % lower (p≤ 0.05). This separation of the growth curves continued such that by Week 14 males given 300 or 1000 mg/kg/day weighed 6 % and 11 % (p≤ 0.05), less than the Controls, with corresponding reductions in overall body weight gain compared with Controls of 12 % (p≤ 0.05) and 21 % (p≤ 0.01) over the duration of the study.

There was no effect on body weight for males given 100 mg/kg/day or for females.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
There was no adverse effect on food intake in either sex at any dose level. Although there were occasional weeks where the food intake of males given o-xylene was statistically significantly higher than that of the Controls, this clearly reflected an unusual pattern of variation in the Controls. Whereas Controls had some weeks of much lower food intake, male groups given o-xylene maintained a more consistent pattern of food intake overall.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
no effects observed
Description (incidence and severity):
There were no ocular changes or abnormalities that were considered to be related to administration of o-xylene.
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
There were no effects on haematology or coagulation parameters that were considered to be test item related. Although a number of parameters showed statistically significant differences between Control and treated groups the differences were minor, not consistent between sexes, lacked a strict dose-response relationship or biological plausibility and so were considered to reflect normal biological variation or to be fortuitous. Mean cell haemoglobin concentration (MCHC) was negligibly (1 %), but statistically significantly lower (p≤ 0.05) in males given 1000 mg/kg/day than in Controls. However, as most values were within the historical range and there were no changes in the primary red cell variates (red blood cell count, haemoglobin concentration, packed cell volume), this trivial difference in this derived parameter was considered not to be meaningful. Group mean platelet count for males given 1000 mg/kg/day was marginally (11 %), but statistically significantly higher than in Controls (p≤ 0.05) and group mean mean platelet volume (MPV) for males given 300 or 1000 mg/kg/day (p≤ 0.05) and plateletcrit (PCT) for males given 1000 mg/kg/day (p≤ 0.01) were also statistically significantly higher than in Controls. Individual platelet counts were within the historical Control range. Although, MPV and PCT values for all animals, including the Controls were outside the historical range, based on the magnitude of differences, which was marginal to negligible, these findings were considered not to be test item related. Group mean neutrophil counts were statistically significantly (p≤ 0.05) higher for males given 300 or 1000 mg/kg/day, however, this was not dose-related, individual values were within historical Control values and as there were no effects on other white blood cell parameters, these differences were considered not to be test item related. Activated partial thromboplastin time (APTT) was marginally (around one second), but significantly (p≤ 0.05) lower, relative to Controls in all male groups given o-xylene. Prothrombin time was marginally (up to 1.5 seconds), but significantly (p≤ 0.05), lower in males given 300 or 1000 mg/kg/day. Since apparent differences in these clotting times were no greater at 1000 mg/kg/day than at 300 mg/kg/day, all individual values were within the historical Control range and, as an increase in these parameters would normally be sought for evidence of impairment of the intrinsic or extrinsic clotting pathways, these variations were considered not to be meaningful. Females given 1000 mg/kg/day had slightly higher red blood cell count (p≤ 0.05) and packed cell volume (p≤ 0.01) than Controls and a lower MCHC (p≤ 0.05). As decreases in primary red cell variates would be more of a concern, and as values remained within normal range, these variations are considered likely to be fortuitous. Group mean total white blood cell count in females given 1000 mg/kg/day was statistically significantly higher (p≤ 0.05) than in Controls as a result of an increase in lymphocyte count (p≤ 0.01). Therefore, as all individual values were well within historical Control ranges these differences are considered of no consequence. Group mean platelet distribution width (PDW) was statistically significantly higher (p≤ 0.01) for females given 1000 mg/kg/day and there was a non-dose related lowering in PCT (p≤ 0.05) for females given o-xylene when compared with Controls. However, as there was no difference in platelet count, these minor differences in these derived indices are considered not to be test item related.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
Plasma albumin concentration in males and females given 1000 mg/kg/day was slightly higher than the Control means (p≤ 0.001), leading to increases in the albumin/globulin ratio for these animals (p≤ 0.01 for males and p≤ 0.001 for females). All of the individual albumin values were within the historical background ranges and this effect was considered to be non-adverse given that a reduction would more normally be sought as evidence of impaired hepatic protein synthesis or of reduced dietary protein intake. Statistically significant inter-group differences in globulin concentrations between females given test item and those given vehicle alone were due to unusually high (above the upper limit of the historical background range) individual values in the Control group. Other statistically significant inter-group differences were minor, not consistent between sexes or lacked a strict dose relationship and so were considered to either reflect normal biological variation or to be fortuitous. These are detailed below:
Plasma sodium was negligibly (<2 %), but significantly, (p≤ 0.01 to p≤ 0.001), higher in males given 300 or 1000 mg/kg/day than in Controls and plasma chloride was negligibly (<1 %), but significantly (p≤ 0.05) lower in males given 1000 mg/kg/day than in Controls. These trivial differences within the normal range were considered to be fortuitous. Males given 1000 mg/kg/day had a higher group mean alanine aminotransferase (ALT) activity (p≤ 0.05) than Controls, with five out of ten individual values above the historical background range. However, in the absence of degenerative pathology changes in the liver, this increased activity was considered to be non-adverse. The slight increase in plasma ALT in males given 1000 mg/kg/day may reflect the observed hepatic hypertrophy in this group. Glucose was lower in a non-dose related manner in males given o-xylene when compared with Controls (p≤ 0.01). Plasma urea was lower in a non-dose related manner for females given o-xylene (p≤ 0.05 to p≤ 0.01), relative to Controls. Cholesterol (p≤ 0.05) and high density lipoproteins (HDL) (p≤ 0.01) were higher in females given 1000 mg/kg/day than in Controls. These differences are considered likely to be entirely fortuitous as they either lacked a dose
relationship, were very minor in degree or, for urea, moved in the wrong direction for diagnostic evidence of renal dysfunction.
Urinalysis findings:
not examined
Behaviour (functional findings):
effects observed, treatment-related
Description (incidence and severity):
There were no effects on sensory reaction or grip strength elicited by o-xylene. During the weekly functional observations, in the open arena, males given 1000 mg/kg/day exhibited effects on arousal (hypoactivity or hyperactivity), unusual posture and abnormal gait. Twitches were observed in one male given 300 mg/kg/day and one male given 1000 mg/kg/day. Increased episodes of arousal in the arena and abnormal gait were observed between Days 21 to 63 and Days 21 to 56 of dosing, respectively. Excessive salivation in the arena was also noted between Days 14 and 70 of dosing for males given 300 or 1000 mg/kg/day, with incidence generally increasing with time. Excessive rearing was noted in a small number of males given 1000 mg/kg/day but this was also seen in one male given 100 mg/kg/day and one Control male and therefore considered not to be test item related. A few females given 300 or 1000 mg/kg/day salivated on removal from the cage between Days 14 to 42. Females given 1000 mg/kg/day also exhibited effects on arousal in the arena (both hyperactivity and hypoactivity), unusual flattened posture, abnormal gait and salivation. In addition, a small number of females exhibited twitching or excessive rearing. Although incidences generally increased with time similarly to the males, these effects were not seen after Day 77 of dosing. During the last week of treatment when the functional observations were performed before and after dosing three females given 1000 mg/kg/day had abnormal gait after dosing, that had not been seen before dosing. In males given 100, 300 or 1000 mg/kg/day o-xylene, movement counts and distance travelled were statistically significantly lower than in Controls (p≤ 0.05 to p≤ 0.01) during a number of post dose counting periods and this was, on occasions, accompanied by a statistically significant increase in time at rest (p≤ 0.05 to p≤ 0.01). There were fewer instances where there were similar statistically significant effects on movement in females and these effects were not dose-related.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Body weight related liver weights for both sexes given 300 or 1000 mg/kg/day (p≤ 0.05 to p≤ 0.001) and females given 100 mg/kg/day (p≤ 0.05) were statistically significantly higher than Controls. Most of the individual relative liver weights for males and females given 1000 mg/kg/day were also above the upper limit of the historical background range. When adjusted for brain weight liver weight was 19 % heavier than Controls in males and 40 % heavier in females (p≤ 0.001) given 1000 mg/kg/day.
Body weight related kidney weights for both sexes given 300 or 1000 mg/kg/day (p≤ 0.01 to p≤ 0.001) and females given 100 mg/kg/day (p≤ 0.05) were statistically significantly higher than Controls. Most of the individual weights were within the historical background range. When adjusted for brain weight kidney weights for both sexes given 300 mg/kg/day were 15 % and 7 % respectively, heavier than those of Controls (p≤ 0.01) and 26 % and 17 % respectively, heavier at 1000 mg/kg/day (p≤ 0.001). Thyroid weights (relative to body weight or brain weight) of females given 1000 mg/kg/day were all heavier than those of Controls (p≤ 0.01 to p≤ 0.001), however, most of the individual weights were within the historical background range.
The adrenals (relative but not absolute weights), of males given 1000 mg/kg/day were 19 % heavier than those of Controls (p≤ 0.05). However, there were no macroscopic or microscopic findings in the adrenals and as all individual values were within the historical control range this increase in adrenal weight was considered not to be adverse. The remaining organ weight differences for relative brain, heart, testes and epididymis weights between Controls and males given up to 1000 mg/kg/day were considered to be due to the slight growth retardation which was evident by the reduced body weight gains over the duration of the study.
Gross pathological findings:
no effects observed
Description (incidence and severity):
A variety of spontaneous changes was noted in Control and animals given o-xylene with no indication of an effect of treatment. The spectrum of these findings is generally consistent with changes encountered in rats of this age kept under laboratory conditions.
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Findings considered to be related to treatment were seen in the liver (hepatocellular hypertrophy) and thyroid (follicular cell hypertrophy) and in the kidneys (eosinophilic inclusions (hyaline droplets) in the tubular epithelium (males only)). Although there were indications of positive α2μ-globulin staining in 3/10 males given 1000 mg/kg/day, there was also minimal staining in 2/10 Controls, thus failing to conclusively indicate that the increased number of hyaline droplets was related to α2μ-globulin. Thus, the mechanism of the increased incidence/severity of hyaline droplets seen within the cortical tubules of male kidneys in this study remained unestablished. A variety of other spontaneous changes was noted in Control and treated animals with no indication of an effect of treatment. The spectrum of these findings is generally consistent with changes encountered in rats of this age kept under laboratory conditions.
Histopathological findings: neoplastic:
no effects observed
Other effects:
effects observed, non-treatment-related
Description (incidence and severity):
Thyroid Hormone Analysis

As samples were obtained at only a single timepoint, these findings represent only a snapshot of thyroid hormone levels. In addition, due to the relatively small group size (n=10), the analysis lacks sufficient statistical power. There was some variation in thyroid hormones within groups with no evidence of a consistent pattern of change elicited by o-xylene. There were no statistically significant inter-group differences for TSH that showed a clear dose response relationship and there were no statistically differences for triiodothyronine (T3) for either sex at any dose. T4 concentrations in females given 1000 mg/kg/day was actually slightly (25 %), but significantly, (p< 0.05) higher than that in Controls but within normal biological variability and, T4 in males given 1000 mg/kg/day was marginally (10 %) lower than in Controls, this difference did not attain statistical significance compared to Controls, nor did it show a dose response relationship. Any of the changes in circulating T4 concentrations
were considered to be within normal biological variability and likely due to the natural diurnal variation in thyroid hormone concentrations.

Oestrous cycles

There was no effect of o-xylene on the oestrous cycles; the data were variable with the majority of females in all groups being in dioestrus.
Key result
Dose descriptor:
NOAEL
Effect level:
300 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
clinical signs
Key result
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
clinical signs
body weight and weight gain
behaviour (functional findings)
organ weights and organ / body weight ratios
histopathology: non-neoplastic
Key result
Critical effects observed:
yes
Lowest effective dose / conc.:
300 mg/kg bw/day (nominal)
System:
urinary
Organ:
kidney
Treatment related:
yes
Dose response relationship:
yes
Relevant for humans:
no

Table 1. body weights (g) – group mean values

Sex: Male

body weight (week number)

Group

1 (#)

2

3

4

5

6

7

Group 1

Control

0 mg/kg/day

Mean

192.5

232.3

265.9

292.1

315.6

335.4

349.1

SD

11.1

14.9

17.6

19.8

22.2

24.8

25.5

N

10

10

10

10

10

10

10

Trend

-

-

-

-

-

-

Group 2

o-xylene

100 mg/kg/day

Mean

194.2

234.3

265.7

290.1

309.3

327.2

341.7

SD

14.4

19.1

22.8

25.6

29.6

31.8

32.3

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

193.2

231.9

259.8

282.1

299.9

314.9

328.7

SD

13.6

16.4

16.4

19.2

21.6

23.4

24.4

N

10

10

10

10

10

10

10

Trend

-

-

-

-

-

-

-

Group 4

o-xylene

1000 mg/kg/day

Mean

195.5

231.5

261.7

282.6

299.9

313.5

324.8

SD

15.5

16.5

19.4

25.4

33.2

35.9

34.7

N

10

10

10

10

10

10

10

Trend

> 0.05

-

-

-

-

-

-

(#) – Williams, Anova & Dunnett

 

Table 1. body weights (g) – group mean values (Continued)

Sex: Male

body weight (week number)

Group

8

9

10

11

12

13

14 (#)

Group 1

Control

0 mg/kg/day

Mean

362.8

375.6

388.1

397.1

404.6

413.2

420.2

SD

27.6

29.8

31.5

32.7

32.4

32.5

35.2

N

10

10

10

10

10

10

10

Trend

-

0

-

-

-

-

Group 2

o-xylene

100 mg/kg/day

Mean

354.8

366.2

378.4

388.5

396.4

405.3

414.3

SD

33.6

36.1

37.4

36.5

37.3

38.4

39.8

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

342.0

352.3

360.9

371.0

377.8

386.5

393.8

SD

25.0

27.4

29.7

32.5

32.1

34.5

33.0

N

10

10

10

10

10

10

10

Trend

-

-

-

-

-

-

> 0.05

Group 4

o-xylene

1000 mg/kg/day

Mean

336.8

344.4

351.3

360.2

366.4

372.4

375.7

SD

38.3

38.8

39.3

41.1

41.6

42.1

41.6

N

10

10

10

10

10

10

10

Trend

-

-

-

-

-

-

≤ 0.05 *

(#) – Williams, Anova & Dunnett: * = p0.05

Table 1. body weights (g) – group mean values (Continued)

Sex: Female

body weight (week number)

Group

1 (#)

2

3

4

5

6

7

Group 1

Control

0 mg/kg/day

Mean

160.4

178.4

189.5

199.4

210.1

220.3

224.5

SD

14.8

14.6

13.9

15.3

15.4

15.5

15.1

N

10

10

10

10

10

10

10

Trend

-

-

-

-

-

-

Group 2

o-xylene

100 mg/kg/day

Mean

159.4

177.1

186.8

198.9

208.8

216.4

221.2

SD

10.8

12.5

13.6

13.8

12.3

12.4

14.7

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

161.7

178.8

189.6

199.5

208.8

217.1

223.7

SD

13.0

16.1

15.4

17.3

15.0

17.2

18.5

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

160.4

181.9

192.3

205.2

214.5

224.3

226.6

SD

13.0

15.0

16.5

18.9

20.1

21.6

23.3

N

10

10

10

10

10

10

10

Trend

> 0 .05

-

-

-

-

-

-

(#) – Williams, Anova & Dunnett

 

Table 1. body weights (g) – group mean values (Continued)

Sex: Female

body weight (week number)

Group

8

9

10

11

12

13

14 (#)

Group 1

Control

0 mg/kg/day

Mean

228.0

231.6

237.0

239.6

239.9

241.3

247.7

SD

18.9

17.6

19.3

18.0

18.8

19.0

19.2

N

10

10

10

10

10

10

10

Trend

-

-

-

-

-

-

Group 2

o-xylene

100 mg/kg/day

Mean

227.4

232.5

236.7

239.2

240.2

244.0

246.7

SD

13.7

10.1

12.1

13.7

14.5

12.2

13.4

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

225.3

232.5

234.6

238.1

239.0

241.5

245.2

SD

18.9

10.1

21.2

21.0

22.3

21.1

23.8

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

231.3

238.5

241.2

242.2

244.4

249.4

249.0

SD

21.5

21.8

22.1

23.1

23.4

23.7

25.9

N

10

10

10

10

10

10

10

Trend

-

-

-

-

-

-

> 0.05

(#) – Williams, Anova & Dunnett

Table 2. body weight gains (g) – group mean values

Sex: Male

body weight (week number)

 

Group

1 to 5 (#)

5 to 14 (#)

1 to 14 (#)

Group 1

Control

0 mg/kg/day

Mean

123.1

104.6

227.7

SD

15.8

17.0

30.1

N

10

10

10

Trend

Group 2

o-xylene

100 mg/kg/day

Mean

115.1

105.0

220.1

SD

21.5

16.8

35.9

N

10

10

10

Trend

> 0.05

-

> 0.05

Group 3

o-xylene

300 mg/kg/day

Mean

106.7

93.9

200.6

SD

15.1

18.0

30.3

N

10

10

10

Trend

≤ 0.05*

> 0.05

≤ 0.05*

Group 4

o-xylene

1000 mg/kg/day

Mean

104.4

75.8

180.2

SD

24.1

12.4

33.3

N

10

10

10

Trend

≤ 0.05*

≤ 0.001***

≤0.01**

(#) – Williams, Anova & Dunnett: *= p0.05, **= p≤ 0.01, ***=p≤ 0.001

 

Table 2. body weight gains (g) – group mean values (Continued)

Sex: Female

body weight (week number)

 

Group

1 to 5 (#)

5 to 14 (#)

1 to 14 (#)

Group 1

Control

0 mg/kg/day

Mean

49.7

37.6

87.3

SD

3.6

5.9

5.7

N

10

10

10

Trend

Group 2

o-xylene

100 mg/kg/day

Mean

49.3

38.0

87.3

SD

6.1

5.8

9.9

N

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

47.1

36.4

83.5

SD

7.2

10.4

14.4

N

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

54.1

34.5

88.6

SD

8.4

8.8

14.0

N

10

10

10

Trend

> 0.05

> 0.05

> 0.05

(#) – Williams, Anova & Dunnett

Table 3. Food intake (g/animal/day) – group mean cage values

Sex: Male

Food Intake Per Animal / Day (g) (Week Number)

Group

1 to 2 (#)

2 to 3 (#)

3 to 4 (#)

4 to 5 (#)

5 to 6 (#)

6 to 7 (#)

7 to 8 (#)

Group 1

Control

0 mg/kg/day

Mean

22.2

22.8

22.1

21.4

20.5

19.7

19.5

SD

1.4

0.8

0.8

1.5

1.1

1.3

1.2

N

4

4

4

4

4

4

4

Trend

Group 2

o-xylene

100 mg/kg/day

Mean

22.9

22.8

21.6

21.1

20.7

20.7

20.7

SD

1.1

1.6

1.0

1.4

0.6

0.3

0.3

N

4

4

4

4

4

1

4

Trend

-

-

-

-

-

> 0.05

> 0.05

Group 3

o-xylene

300 mg/kg/day

Mean

22.6

22.7

21.8

21.2

21.0

21.4

21.1

SD

0.6

0.8

1.4

1.1

0.9

0.7

0.5

N

4

4

4

4

4

4

1

Trend

-

-

-

-

-

≤ 0.05*

≤ 0.05*

Group 4

o-xylene

1000 mg/kg/day

Mean

22.1

24.3

22.6

22.4

22.3

22.0

21.5

SD

1.3

2.4

2.3

3.1

2.9

1.4

1.6

N

4

4

4

4

4

4

4

Trend

> 0.05

> 0.05

> 0.05

> 0.05

> 0.05

≤ 0.05*

≤ 0.05*

(#) – Williams, Anova & Dunnett: * = p0.05

 

Table 3. Food intake (g/animal/day) – group mean cage values (Continued)

Sex: Male

Food Intake Per Animal / Day (g) (Week Number)

Group

8 to 9 (#)

9 to 10 (#)

10 to 11 (#)

11 to 12 (#)

12 to 13 (#1)

13 to 14 (#)

1 to 14 (#)

Group 1

Control

0 mg/kg/day

Mean

19.1

18.9

18.3

17.8

18.1

17.2

19.8

SD

1.6

1.6

1.3

1.2

1.7

1.3

1.2

N

4

4

4

4

4

4

4

Trend

Group 2

o-xylene

100 mg/kg/day

Mean

20.3

20.2

20.3

20.3

21.1

19.1

20.9

SD

0.6

0.7

0.4

0.3

0.2

1.2

0.6

N

4

4

4

4

4

4

4

Trend

-

-

≤ 0.05*

≤ 0.05*

≤ 0.05*

-

-

Group 3

o-xylene

300 mg/kg/day

Mean

20.6

20.3

20.7

20.5

20.0

19.0

21.0

SD

1.0

0.9

0.6

0.6

0.3

0.4

0.6

N

4

4

4

4

4

4

1

Trend

> 0.05

-

≤ 0.05*

≤ 0.05*

≤ 0.05*

> 0.05

-

Group 4

o-xylene

1000 mg/kg/day

Mean

21.3

20.6

20.9

21.5

21.3

19.7

21.7

SD

1.6

1.8

2.1

2.9

2.4

2.4

2.0

N

4

4

4

4

4

4

4

Trend

≤ 0.05*

> 0.05

≤ 0.05*

≤ 0.05*

≤ 0.05*

≤ 0.05*

> 0.05

(#) – Williams, Anova & Dunnett: * = p0.05

(#1) – Shirley, Kruskal-Wallis & Steel: * = p0.05

Table 3. Food intake (g/animal/day) – group mean cage values (Continued)

Sex: Female

Food Intake Per Animal / Day (g) (Week Number)

Group

1 to 2 (#)

2 to 3 (#)

3 to 4 (#)

4 to 5 (#)

5 to 6 (#)

6 to 7 (#)

7 to 8 (#)

Group 1

Control

0 mg/kg/day

Mean

14.8 n

14.3 n

15.1 n

15.1 n

15.2 n

14.1 n

14.3 n

SD

1.4

0.2

0.5

0.7

0.6

0.6

0.6

N

2

2

2

2

2

2

2

Group 2

o-xylene

100 mg/kg/day

Mean

14.3 n

14.1 n

14.6 n

14.5 n

14.6 n

14.0 n

14.2 n

SD

1.2

0.9

1.2

1.3

1.4

1.7

1.6

N

2

2

2

2

2

2

2

Group 3

o-xylene

300 mg/kg/day

Mean

14.4 n

14.5 n

15.0 n

15.1 n

14.9 n

14.2 n

14.1

SD

0.7

0.6

0.8

0.9

1.2

0.9

1.2

N

2

2

2

2

2

2

2

Group 4

o-xylene

1000 mg/kg/day

Mean

13.7 n

15.4 n

15.7 n

15.3 n

15.4 n

14.6 n

14.8 n

SD

0.0

0.6

0.5

0.5

0.6

0.2

0.2

N

2

2

2

2

2

2

2

(#) – Shirley, Kruskal-Wallis & Steel: n – inappropriate for statistics

 

Table 3. Food intake (g/animal/day) – group mean cage values (Continued)

Sex: Female

Food Intake Per Animal / Day (g) (Week Number)

Group

8 to 9 (#)

9 to 10 (#)

10 to 11 (#)

11 to 12 (#)

12 to 13 (#)

13 to 14 (#)

1 to 14 (#)

Group 1

Control

0 mg/kg/day

Mean

13.9 n

13.9 n

13.3 n

13.2 n

13.6 n

12.2 n

14.1 n

SD

0.4

0.6

0.8

0.9

1.3

0.5

0.6

N

2

2

2

2

22

2

2

Group 2

o-xylene

100 mg/kg/day

Mean

13.8 n

14.1 n

13.2 n

13.2 n

12.9 n

12.2 n

13.8 n

SD

1.2

1.1

1.9

1.9

1.1

2.0

1.4

N

2

2

2

2

2

2

2

Group 3

o-xylene

300 mg/kg/day

Mean

14.0 n

14.0 n

13.9 n

13.7 n

13.4 n

12.6 n

14.1 n

SD

1.0

1.1

1.1

1.5

1.4

1.2

1.0

N

2

2

2

2

2

2

2

Group 4

o-xylene

1000 mg/kg/day

Mean

14.6 n

14.5 n

14.0 n

14.2 n

13.2 n

12.8 n

14.5 n

SD

0.0

0.6

0.3

0.3

0.5

0.0

0.1

N

2

2

2

2

2

2

2

(#) – Shirley, Kruskal-Wallis & Steel: n – inappropriate for statistics

Table 4. Thyroid analysis – group mean values

Sex: Male

Mean Total

T3 (ng/mL)

Mean Total

T4 (ng/mL)

Mean Total

TSH (ng/mL)

Group

(#)

(#)

(#1)

Group 1

Control

0 mg/kg/day

Mean

16.639

427.735

0.498

SD

2.676

82.494

0.210

N

10

10

10

Trend

Group 2

o-xylene

100 mg/kg/day

Mean

16.682

440.194

0.510

SD

5.221

46.662

0.152

N

9

9

9

Group 3

o-xylene

300 mg/kg/day

Mean

19.031

437.204

0.555

SD

5.116

89.539

0.192

N

9

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

18.468

383.893

0.605

SD

4.758

44.318

0.314

N

9

10

10

Trend

> 0.05

> 0.05

> 0.05

(#) – Williams, Anova & Dunnett

(#1) – Shirley, Kruskal-Wallis & Steel

 

Table 4. Thyroid analysis – group mean values (Continued)

Sex: Male

Mean Total

T3 (ng/mL)

Mean Total

T4 (ng/mL)

Mean Total

TSH (ng/mL)

Group

(#)

(#)

(#1)

Group 1

Control

0 mg/kg/day

Mean

17.233*

277.065

0.673*

SD

5.928

37.594

0.303

N

8

10

10

Trend

Group 2

o-xylene

100 mg/kg/day

Mean

12.174

291.000

0.497

SD

3.053

71.508

0.182

N

8

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

14.009

296.776

0.421*

SD

3.301

59.956

0.066

N

9

10

10

Trend

-

> 0.05

-

Group 4

o-xylene

1000 mg/kg/day

Mean

19.130

346.589

0.595

SD

6.317

56.001

0.209

N

10

10

10

Trend

> 0.05

≤ 0.05*

> 0.05

(#) – Williams, Anova & Dunnett: * = p≤ 0.05

(#1) – Williams, Anova & Dunnett(Log): * = p0.05

Table 5. Functional observation battery – rodents – key values

HOME CAGE

ON REMOVAL FROM CAGE

PERFORMANCE IN OPEN FIELD

RESPONSE TO SENSORIMOTOR STIMULI

Arousal upon openingcage

0 = normal

1 = increased

2 = decreased

 

Dispersion in cage

0 = normal

1 = closely bunched

2 = loosely bunched

 

Excessive vocalisation

0 = absent

1 = present

 

Fighting

0 = absent

1 = present

 

Stereotypical behaviour

0 = absent

1 = present

 

Aggressive behaviour

0 = absent

1 = present

 

Convulsions

0 = absent

1 = short

2 = sustained

Reaction to handling

0 = normal

1 = no resistance

2 = vocalising

3 = tense

4 = squirming

 

Lacrimation

0 = normal

1 = excess at eyelid margin

2 = extends beyond margin

 

Salivation

0 = normal

1 = slight

2 = marked

 

Chromodacryorrhea

0 = absent

1 = present

 

Exophthalmos

0 = absent

1= present

 

Arousal in arena

0 = normal

1 = hyperactive

2 = hypoactive

 

Unusual posture

0 = absent

1 = hunched

2 = flattened

 

Abnormal gait

0 = absent

1 = slight

2 = marked

 

Convulsions arena

0 = absent

1 = slight

2 = marked

 

Tremors

0 = none

1 = intermittent

2 = constant

 

Twitches

0 = normal

1 = slight

2 = marked

 

Urination

0 = none

1 = present

2 = excess

Defecation

0 = none

1 = present

2 = soft/liquid faeces

 

Excessive vocalisation

0 = absent

1 = present

 

Piloerection

0 = normal

1 = roughened fur

2 = piloerection

 

Palpebral closure

0 = normal

1 = partially closed

2 = shut

 

Respiration

0 = normal

1 = rapid

2 = slow

3 = laboured

 

Straub tail

0 = absent

1 = present

 

Salivation

0 = absent

1 = present

2 = excessive

 

Excessive rearing

0 = absent

1 = present

Approach response

0 = present

1 = absent

2 = excessive

 

Touch response

0 = present

1 = absent

2 = excessive

 

Aud Startle response

0 = present

1 = absent

2 = excessive

 

Tail pinch response

0 = present

1 = absent

2 = excessive

 

Forelimb reflex

0 = early extension

1 = extension after contact

2 = no extension

 

Righting reflex

0 = present

1 = slow

2 = absent

 

Table 5. Functional observation battery – home and arena – group mean values

Sex: Male

Arousal open cage (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

 

Table 5. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Arousal open cage (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

Table 5.2. Functional observation battery – home and arena – group mean values (Continued)

Sex: Male

Dispersion in cage (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

12

0

0

0

0

0

0

SD

3

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Dispersion in cage (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Fighting (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Fighting (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Excessive vocalisation cage (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Excessive vocalisation cage (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Stereotyped behaviour (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Stereotyped behaviour (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

Tabl e 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Aggressive behaviour (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Aggressive behaviour (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Convulsions cage (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Convulsions cage (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Reacting to handling (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Reacting to handling (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Lacrimation (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Lacrimation (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Salivation (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Salivation (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Chromodacryorrhea (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Chromodacryorrhea (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Exophthalmos (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Exophthalmos (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Arousal in arena (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

4

0

SD

0

0

0

0

0

2

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

2

0

0

0

0

SD

0

0

2

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

4

0

1

2

SD

0

0

0

2

0

1

2

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Arousal in arena (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

1

0

0

1

0

0

SD

1

0

0

1

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

2

0

0

0

0

0

SD

2

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

5

0

3

0

0

0

SD

2

0

2

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Unusual posture (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

4

0

SD

0

0

0

0

0

2

0

N

10

10

10

10

10

10

10

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Unusual posture (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

4

0

0

0

SD

0

0

2

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Abnormal gait (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

1

0

1

0

SD

0

0

0

1

0

1

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Abnormal gait (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

1

0

0

0

0

0

SD

1

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

1

1

0

0

0

0

SD

1

1

0

0

0

0

N

10

10

10

10

10

10

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Convulsions arena (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Convulsions arena (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Tremors (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Tremors (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Twitches (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

1

0

SD

0

0

0

0

0

1

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Twitches (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

1

0

0

0

0

0

SD

1

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Excessive vocalisation arena (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Excessive vocalisation arena (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Piloerection (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Piloerection (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Palpebral closure (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Palpebral closure (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Respiration (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Respiration (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Straub tail (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Straub tail (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Salivation arena (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

6

4

0

3

0

SD

0

0

1

1

0

2

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Salivation arena (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

1

0

0

2

0

0

SD

1

0

0

1

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

1

3

1

0

0

0

SD

1

2

1

0

0

0

N

10

10

10

10

10

10

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Excessive rearing (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

1

0

1

0

SD

0

0

0

1

0

1

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Excessive rearing (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

1

0

0

SD

0

0

0

1

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

1

0

0

SD

0

0

0

1

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

1

0

0

0

0

0

SD

1

0

0

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Urination (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

5

1

0

7

2

3

5

SD

1

1

0

1

1

1

1

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

5

0

1

6

5

3

5

SD

1

0

1

1

2

1

1

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

3

2

0

6

6

6

4

SD

1

1

0

1

2

1

1

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

2

0

0

7

9

3

3

SD

1

0

0

1

2

1

1

N

10

10

10

10

10

10

10

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Urination (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

4

3

1

4

5

7

SD

1

1

1

1

1

1

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

2

4

4

2

5

4

SD

1

1

1

1

1

1

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

1

4

7

3

6

5

SD

1

2

2

1

1

1

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

6

6

4

2

2

8

SD

2

2

1

1

1

1

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Defecation (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

2

0

1

0

0

0

0

SD

1

0

1

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

2

1

0

0

0

0

1

SD

1

1

0

0

0

0

1

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Male

Defecation (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Female

Arousal open cage (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Female

Arousal open cage (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Female

Dispersion in cage (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Female

Dispersion in cage (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Female

Fighting (Day number)

Group

-1

7

14

21

28

35

42

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

Group 4

o-xylene

1000 mg/kg/day

Mean

0

0

0

0

0

0

0

SD

0

0

0

0

0

0

0

N

10

10

10

10

10

10

10

 

 

Table 5.2. Functional observation battery – home and arena – group mean values (continued)

Sex: Female

Fighting (Day number)

Group

49

56

63

70

77

84

Group 1

Control

0 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 2

o-xylene

100 mg/kg/day

Mean

0

0

0

0

0

0

SD

0

0

0

0

0

0

N

10

10

10

10

10

10

Group 3

o-xylene

300 mg/kg/day

Mean

0

0

0

0

0

0