Registration Dossier

Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
GLP compliance:
yes (incl. QA statement)
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Details on test material:
As per IUCLID Sections 1.1. 1.2. and 4.1.

Test animals

Species:
rat
Strain:
Wistar
Details on species / strain selection:
Species: Rat
Strain: Wistar Han™
Sub-strain: RccHan™: WIST
Source of supply: Envigo RMS (UK) Ltd., Oxon, UK
Number: 80
Sex: Males and females
Age: Approximately 6 to 8 weeks at start of treatment
Other: Weight variation must not exceed ± 20% of the mean weight for either sex at start of treatment
Sex:
male/female
Details on test animals or test system and environmental conditions:
Environment
Environmental conditions will be maintained within the following target parameters:
Target temperature: 22 ± 3 °C
Target humidity: 30 to 70%
Lighting: Twelve hours of continuous artificial light in each twenty-four hour period
Ventilation: At least fifteen air changes per hour
Animals are housed in accordance with the 'Code of Practice for the Housing and Care of Animals Used in Scientific Procedures' and in alignment with the United Kingdom Home Office recommendations and requirements.

Housing
Animals are gang-housed in groups of up to four by sex in polypropylene cages with stainless steel mesh lids and furnished with softwood flake bedding (Datesand Ltd., Cheshire, UK). Certificates of analysis for the bedding will be provided by the supplier. The bedding is considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.

Diet and Water
The animals are provided with Rodent 20 l4C Teklad Global Certified Pelleted Diet (Envigo RMS (UK) Ltd., Oxon, UK) and tap water ad libitum.
Drinking water is routinely analyzed (at least annually) and certificates of analysis for diet are provided by the supplier.
The diet and drinking water are considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.

Environmental Enrichment
The animals are provided with environmental enrichment items: wooden chew blocks and cardboard fun tunnels (Datesand Ltd., Cheshire, UK) or suitable alternatives. Certificates of analysis for the environmental enrichment items will be provided by the supplier. These enrichment items are considered not to contain any contaminants that could reasonably be expected to affect the purpose or integrity of the study.

Administration / exposure

Route of administration:
oral: gavage
Details on route of administration:
Once daily, by gavage, using a stainless steel dosing cannula attached to a graduated syringe for ninety consecutive days. Dosing is performed at a similar time each day wherever possible.
Vehicle:
polyethylene glycol
Details on oral exposure:
Animals will be allocated to test groups as follows:
Dose groups Group Size Animal Numbers Treatment Necropsy
(mg/kg bw/d) & Sex Males Females period Day
I. Control 10 male & 10 female 1-10 11-20 90 days 91
2. 10 10 male & 10 female 21-30 31-40 90 days 91
3. 100 10 male & 10 female 41-50 51-60 90 days 91
4. 1000 10 male & 10 female 61-70 71-80 90 days 91

Control animals are treated with the vehicle alone over the same treatment period and at the same dose volume as that of test animals.
The volume oftest and control item administered to each animal is be based on the most recent scheduled body weight and adjusted accordingly.
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
90 days
Frequency of treatment:
7 days a week
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (actual dose received)
Dose / conc.:
10 mg/kg bw/day (actual dose received)
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle

Examinations

Observations and examinations performed and frequency:
Serial Observations

1 General ObservationslMeasurements

1.1 MorbiditylMortality Inspection
Twice daily, early and late during the working period.

1.2 Clinical Observations
Individual clinical observations are performed immediately before dosing, up to 30 minutes after dosing and one hour after dosing. All observations are recorded.

1.3 Body Weights
Individual body weights are recorded on Day 1 (prior to dosing) and at weekly intervals thereafter. Individual body weights are recorded at terminal kill.

1.4 Food Consumption
Dietary intake are recorded weekly for each cage group. Weekly food efficiency (body weight gain/food intake) is calculated.

1.5 Water Consumption
Water consumption is monitored daily by visual inspection of water bottles. Measurement is initiated if a treatment-related effect is suspected, at the discretion of the Study Director.

2 Specialist Evaluations

2.1 Functional Observations
Detailed individual clinical observations are performed on all test and control group animals before the first exposure to the test item and once weekly thereafter. These observations are performed outside the home cage, in a standard arena, at least two hours after dosing (where applicable) to ensure that any transient effects of treatment are identified. All observations are recorded. In addition, sensory reactivity to different stimuli (auditory, visual and proprioceptive), grip strength (fore and hind limb) and motor activity (using a 44 photobeam unit) are measured once during Week 12 of the study, at least two hours after dosing, whenever possible.

2.2 Ophthalmoscopy
Ophthalmoscopic examination is performed on all animals before the start of treatment and all control and high dose animals during Week 12 of the study. Examinations includes observation of the anterior structures of the eye and, following pupil dilation, detailed examination of the internal structures of the eye using a ophthalmoscope. Where treatment related changes are suspected in the high dose group at Week 12 examinations, ophthalmoscopic examination is extended to animals of the other dose groups.

3 In-life Sampling and Analysis
Hematological and blood chemical investigations are performed on all test and control group animals at the end of the treatment period (Day 90). Blood samples are withdrawn from the lateral tail vein. Repeat blood samples are withdrawn by cardiac puncture at termination if required. Animals are not fasted prior to blood sampling.

3.1 Hematology
Hemoglobin (Hb)
Erythrocyte count (RBC)
Hematocrit (Hct)
Erythrocyte indices
- mean corpuscular hemoglobin (MCH)
- mean corpuscular volume (MCV)
- mean corpuscular hemoglobin concentration (MCHC)
Total leucocyte count (WBC)
Differential leucocyte count
- neutrophils (Neut)
- lymphocytes (Lymph)
- monocytes (Mono)
- eosinophils (Eos)
- basophils (Bas)
Prothrombin time (CT)
Platelet count (PL T)
Activated partial thromboplastin time (APTT)
Reticulocyte count' (Retic)
'Blood film will be prepared but only examined at Study Director's discretion

3.2 Blood Chemistry
Urea
Glucose
Total protein (Tot.Prot.)
Albumin
Albumin/Globulin (A/G) ratio (by calculation)
Sodium (Na+)
Potassium (K +)
Chloride (CI-)
Calcium (Ca++)
Inorganic phosphorus (P)
Aspartate aminotransferase (ASAT)
Alanine aminotransferase (ALAT)
Alkaline phosphatase (AP)
Creatinine (Creat)
Total cholesterol (Chol)
Total bilirubin (Bili)
Bile acids
Sacrifice and pathology:
Terminal investigations

1 Necropsy
Post mortem procedures are performed on all animals and include any animal found dead or killed in extremis during the study. All animals are killed by intravenous overdose of a suitable barbiturate agent, followed by exsanguination at termination. Selected tissues are weighed and whole or samples of required tissues are retained in 10% Buffered Formalin. The lungs are inflated to approximately normal respiratory volume with buffered 10% formalin before immersion in fixative.

Tissue/Organ Weigh Retain Process &
Examine

Adrenals X X X
Aorta (thoracic) X X
Bone and bone marrow (femur including stifle joint) X
Bone and Bone marrow (sternum) X X
Brain (including cerebrum, cerebellum and pons) X X X
Caecum X X
Colon X X
Duodenum X X
Esophagus X X
Epididymides (retained in Modified Davidson's Fluid) X X X
Eyes (retained in Davidson's Fluid) X X
Gross lesions X X
Heart X X X
Ileum (including peyer's patches) X X
Jejunum X X
Kidnevs X X X
Liver X X X
Lungs (with bronchi) X X
Lymph nodes (mandibular and mesenteric) X X
Mammary gland X X
Muscle (skeletal) X
Ovaries X X X
Pancreas X X
Pituitary X X
Prostate X X
Rectum X X
Salivary glands (submaxillary) X X
Sciatic nerve X X
Seminal vesicles (including coagulating gland) X X
Skin X X
Spinal cord (cervical, mid-thoracic and lumbar) X X
Spleen X X X
Stomach X X
Testes (retained in Modified Davidson's Fluid) X X X
Thymus X X X
ThvroidlParathyroid X X
Tongue X
Trachea X X
Urinary bladder X X
Uterus (with Cervix) X X X
Vagina X X

1.1 Gross Examination
Full external and internal macroscopic examination of all animals.

1.2 Organ Weights
The tissues listed in the table in Section 1 are weighed for all surviving animals at terminal kill. As far as possible, fat and connective tissues are trimmed from each organ and the organs will be weighed fresh prior to being placed in fixative. Paired organs are weighed together and not separately, but individual weights are recorded if abnormal size of one of the paired organs is suspected. Organ weights are not routinely recorded for decedent animals, however they are recorded if the death occurs towards the end of the study and it is considered that the data collected may be valid.

2 Histology
Samples of the tissues preserved in their appropriate fixatives are dispatched to the histology processing Test Site.
Initially:
a) all tissues from animals that die or are killed in extremis during the study
b) all tissues from control and high dose group animals
c) any abnormal tissues from low and intermediate dose groups
are routinely processed to paraffin wax, sectioned and stained with Haematoxylin and Eosin. In addition, sections of testes are stained with Periodic Acid-Schiff (PAS) stain. The examination of gross lesions covers the normal background incidence of macroscopic gross lesions observed for this study design. In some circumstances, the incidence of findings may be increased due to treatment-related effects (e.g. discoloration of tissues due to strongly colored test item). Where the abnormality observed relates to the contents of a tissue (i.e. abnormal contents within parts of the gastro-intestinal tract) which would most probably be lost during the histology processing procedures, then these abnormalities are not normally processed as gross lesions.

3 Pathology
3.1 Histopathology
Prepared slides are sent to the Study Pathologist, whereupon special staining techniques are used, where appropriate. Where suspected treatment-related lesions are seen in the high dose group, histology processing and histopathological examination of the affected tissue(s) may be extended to animals of other dose groups. Where the Study Pathologist is a Principal Investigator working at a Test Site, a peer review
of the histopathology results for the study are conducted by the Test Facility. Where the Study Pathologist is part of the Test Facility, a review of the histopathology results for the study are conducted by an appointed Responsible Scientist working at a designated Test Site. On completion of histopathology evaluations, a histopathology examination phase report is sent to the Study Director by the Study Pathologist and this represents the consensus view of both pathologists.
Statistics:
Where considered appropriate, quantitative data is subjected to statistical analysis to detect the significance of intergroup differences from control; statistical significance is achieved at a level of p<0.05. Analysis is performed on, but not necessarily restricted to, the following parameters:
Body weight change, motor activity, grip strength, hematology, blood chemistry, absolute organ weights and body weight-relative organ weights
Data is analyzed using the decision tree from the Provantis™ Tables and Statistics Module as detailed below:
If appropriate, transformation of the data is performed using the most suitable method.
The homogeneity of variance from mean values is analyzed using Bartlett's test.
Intergroup variance is assessed using suitable ANOVA, or if required, ANCOV A with appropriate covariates. Any transformed data is analyzed to find the lowest treatment level that shows a significant effect, using the Williams Test for parametric data or the Shirley Test for non-parametric data. If no dose response is found, but the data shows nonhomogeneity of means, the data is analyzed by a stepwise Dunnett's (parametric) or Steel (non-parametric) test to determine significant difference from the control group. Where the data is unsuitable for these analyses, then pair-wise tests are performed using the Student t-test (parametric) or the Mann-Whitney U test (non-parametric).
Other statistical analysis pathways may be implemented at the discretion of the Study Director and will be retained within the raw data for the study.
Flags for significance between control and treated groups (* for p<0.05, ** for p<0.01 and, where appropriate, *** for p<0.001) is indicated in the tables.

Results and discussion

Results of examinations

Description (incidence and severity):
A summary incidence of daily clinical observations is given in Table 1. Individual data is presented in Appendix 1.

Increased salivation was evident in animals of either sex treated with 1000 mg/kg bw/day from Day 5 (females) and Day 6 (males) onwards. Increased salivation was also evident in one male treated with 100 mg/kg bw/day on Day 34. Noisy respiration was evident in animals of either sex treated with 1000 mg/kg bw/day between Days 12 and 79 (males) and Days 13 and 44 (females). Noisy respiration was also evident in one male treated with 100 mg/kg bw/day on Day 26 and 76 and in one male treated with 10 mg/kg bw/day on Day 53.

No such effects were detected in females treated with 100 or 10 mg/kg bw/day.

One female treated with 1000 mg/kg bw/day had scab formation between Days 19 and 35. Observations of this nature are not uncommon in gang housed animals and in isolation, was considered unrelated to treatment.
Mortality:
no mortality observed
Description (incidence):
There were no unscheduled deaths.
Description (incidence and severity):
Group mean weekly body weights and standard deviations are given in Table 6 and are presented graphically in Figure 1 and Figure 2. Group mean weekly body weight gains and standard deviations are given in Table 7 (statistically significant differences are indicated). Individual data are given in Appendix 7 and Appendix 8.

Statistically significant reductions (p<0.05-0.01) in body weight gains were evident in males treated with 1000 mg/kg bw/day during Weeks 1, 4, 6, 8, 10, 11, and 12; with actual body weight losses being evident during Week 12. A slight improvement in these males was evident during Weeks 5 and 13; with statistically significant increases in body weight gains being evident during these weeks, however, a 20% reduction in overall body weight gain was evident when compared to controls. Females treated with 1000 mg/kg bw/day also showed a slight reduction (7%) in overall body weight gain. A statistically significant reduction (p<0.05) in body weight gain was evident in these females during Week 10.

No such effects were evident in animals of either sex treated with 100 or 10 mg/kg bw/day.

Males treated with 100 mg/kg bw/day showed a statistically significant increase (p<0.01) in body weight gain during Week 5 whilst females from this treatment group showed a statistically significant reduction (P<0.05) in body weight gain during Week 10. An increase in body weight gain for males is considered not to reflect an adverse effect of treatment and in the absence of a true dose related response in females the intergroup differences were considered not to be of toxicological significance.
Description (incidence and severity):
Group mean weekly food consumptions are given in Table 8 and are presented graphically in Figure 3 and Figure 4. Weekly food efficiencies are given in Table 9.

Males treated with 1000 mg/kg bw/day showed a slight reduction in overall food consumption. Food conversion efficiency was also reduced in these males and generally followed the reductions that were evident in body weight gains.

No such effects were evident in females treated with 1000 mg/kg bw/day or in animals of either sex treated with 100 or 10 mg/kg bw/day.
Description (incidence and severity):
Visual inspection of water bottles did not reveal any intergroup differences.
Description (incidence and severity):
Individual ophthalmoscopic examination findings are given in Appendix 9.

Ophthalmoscopic examination of animals of either sex from the control and 1000 mg/kg bw/day dose groups during Week 12 of the treatment period did not indicate any treatment related differences.
Description (incidence and severity):
Group mean values and standard deviations for test and control group animals are given in Table 10 (statistically significant differences are indicated). Individual data are given in Appendix 10 and Appendix 11.

Males treated with 1000 mg/kg bw/day showed statistically significant reductions (p<0.05-0.01) in hemoglobin, erythrocyte count, hematocrit, mean corpuscular hemoglobin and mean corpuscular volume. Females treated with 1000 mg/kg bw/day showed statistically significant increases (p<0.01) in total leucocyte count and lymphocytes.

No toxicologically significant effects were detected in animals of either sex treated with 100 or 10 mg/kg bw/day.

Males treated with 1000 and 100 mg/kg bw/day showed a statistically significant increase (p<0.05) in neutrophil counts. The majority of individual values were within historical control ranges and in the absence of a true dose related response the intergroup differences were considered not to be of toxicological significance. Females treated with 1000 mg/kg bw/day showed statistically significant reductions (p<0.05-0.01) in mean corpuscular hemoglobin and mean corpuscular volume. All of the individual values were within historical control ranges and in the absence of any associated histopathological changes, the intergroup differences were considered not to be of toxicological importance.
Description (incidence and severity):
Group mean values and standard deviations for test and control group animals are given in Table 11 (statistically significant differences are indicated). Individual data are given in Appendix 12 and Appendix 13.

Animals of either sex treated with 1000 mg/kg bw/day showed statistically significant reductions (p<0.01) in total protein and albumin and a statistically significant increase (p<0.01) in bilirubin. Males treated with 1000 mg/kg bw/day also showed statistically significant increases (p<0.05-0.01) in albumin/globulin ratio, aspartate aminotransferase, cholesterol, calcium concentration, phosphorus and bile acids whilst females from this treatment group also showed a statistically significant increase (p<0.05) in alanine aminotransferase. Females treated with 1000 mg/kg bw/day also showed an increase in bile acids, however, statistical significance was not achieved. Although the majority of the individual values for total protein, albumin (males), albumin/globulin ratio, calcium concentration and phosphorus were within the normal expected ranges, the remaining individual values for asapartate aminotransferase, cholesterol, bilirubin, bile acids, alanine aminotransferase and albumin (females) were outside of historical control ranges and therefore, the potential association with the histopathological changes in the liver means that a relationship to treatment for all of these parameters cannot be excluded.

No toxicologically significant effects were detected in animals of either sex treated with 100 or 10 mg/kg bw/day.

Males from all treatment groups showed a statistically significant increase (p<0.05-0.01) in urea. Males treated with 10 mg/kg bw/day also showed a statistically significant increase in glucose (p<0.05). All of the individual values for urea and the majority of individual values for glucose were within historical control ranges and in the absence of true dose related responses the intergroup differences were considered not to be of toxicological importance.

Males treated with 100 mg/kg bw/day showed a statistically significant increase in cholesterol whilst females from this treatment group showed a statistically significant increase in bilirubin. Although similar effects were evident at 1000 mg/kg bw/day, no histopathological correlates were evident at 100 mg/kg bw/day, therefore, the intergroup differences were considered not to represent an adverse effect of treatment.
Description (incidence and severity):
Functional Observations
A summary incidence of behavioral assessment observations is given in Table 2 and group mean behavioral assessment scores are given in Table 3. Group mean functional performance test values and standard deviations are given in Table 4 (statistically significant differences are indicated). Individual values are given in Appendix 4 and Appendix 5. Group mean sensory reactivity assessments are given in Table 5. Individual responses are given in Appendix 6.

Behavioral Assessments
There were no treatment-related changes in the behavioural parameters measured.

Functional Performance Tests
There were considered to be no toxicologically significant changes in functional performance.

Males from all treatment groups showed a statistically significant reduction (p<0.05) in overall activity. In the absence of a true dose related response or any clinical signs to suggest a possible neurotoxic effect, the intergroup differences were considered not to be of toxicological significance.

Sensory Reactivity Assessments
There were no treatment-related changes in sensory reactivity.
Description (incidence and severity):
Group mean absolute and relative organ weights and standard deviations for test and control group animals are presented in Table 13 (statistically significant differences are indicated). Individual data are given in Appendix 15 and Appendix 16.

Females treated with 1000 mg/kg bw/day showed a statistically significant increase (p<0.01) in liver weight both absolute and relative to terminal body weight. Males treated with 1000 mg/kg bw/day showed a statistically significant increase (p<0.01) in kidney weights both absolute and relative to terminal body weight.

No treatment-related effects were detected in animals of either sex treated with 100 or 10 mg/kg bw/day.
Description (incidence and severity):
A summary incidence of necropsy findings is given in Table 12. Individual data are given in Appendix 14.

Five males treated with 1000 mg/kg bw/day had pale kidneys. Two of these males also had enlarged kidneys and one of these males had enlarged kidneys and a thickened glandular region of the stomach. Another male from this treatment group also had enlarged kidneys without associated pallor.

No toxicologically significant effects were evident in females treated with 1000 mg/kg bw/day or in animals of either sex treated with 100 or 10 mg/kg bw/day.

The following findings were without histopathological correlates and were considered to be incidental and not related to treatment with the test item:

One control male, two control females, one male and two females treated with 10 mg/kg bw/day, one male and two females treated with 100 mg/kg bw/day and one male and two females treated with 1000 mg/kg bw/day had reddened lungs at necropsy. One control female and one male treated with 100 mg/kg bw/day had increased renal pelvic space in one kidney. One control male had a raised limiting ridge.
Description (incidence and severity):
A complete histopathology phase report is presented in Annex 1.

The following treatment related microscopic abnormalities were detected:

Caecum: Mucosal hypertrophy was evident in eight males treated with 1000 mg/kg bw/day. No such effects were evident in females treated with 1000 mg/kg bw/day or in animals of either sex treated with 100 or 10 mg/kg bw/day.

Small Intestine: Lymphangiectasis was evident in the villi of the jejunum or ileum in five males and six females treated with 1000 mg/kg bw/day. No such effects were evident in animals of either sex treated with 100 or 10 mg/kg bw/day.

Kidneys: Multifocal basophilic tubules were evident in eight males and four females treated with 1000 mg/kg bw/day. Tubular dilation was evident in four males and five females from this treatment group and mineralization in the pelvis of the kidney was also evident in five males and females. No such effects were evident in animals of either sex treated with 100 or 10 mg/kg bw/day.

Liver: Minimal or mild centrilobular hypertrophy was evident in five males and seven females treated with 1000 mg/kg bw/day. No such effects were evident in animals of either sex treated with 100 or 10 mg/kg bw/day.

Ovaries: Interstitial cell hyperplasia was evident in nine females treated with 1000 mg/kg bw/day. No such effects were evident in females treated with 100 or 10 mg/kg bw/day.

No other findings were present at histopathology which could be attributed to treatment with the test item.

Effect levels

Key result
Dose descriptor:
NOAEL
Effect level:
100 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical biochemistry
haematology
histopathology: non-neoplastic

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Dose Levels: Group 1 - 0(Control) Group 2 - 10 mg/kg bw/day Group 3 - 100 mg/kg bw/day Group 4 - 1000 mg/kg bw/day

Table 1 Summary Incidence of Daily Clinical Observations

Day Numbers Relative to Start Date

 

 

 

Sex: Male

Group 1

0(Control) bw/day

Group 2

10 mg/kg bw/day

Group 3

100 mg/kg

Group 4

1000 mg/kg bw/day

Scheduled kill

Number of Animals

Days from - to

 

10

91 91

 

10

91 91

 

10

91 91

 

10

91 91

Increased salivation

Number of Animals

Days from - to

 

.

.

 

.

.

 

1

34 34

 

10

6 90

Noisy respiration

Number of Animals

Days from - to

 

.

.

 

1

53 53

 

1

26 76

 

7

12 79

 

Day Numbers Relative to Start Date

 

 

 

Sex: Female

Group 1

0(Control) bw/day

Group 2

10 mg/kg bw/day

Group 3

100 mg/kg

Group 4

1000 mg/kg bw/day

Scab

Number of Animals

Days from - to .

 

.

.

 

.

.

 

.

.

 

1

19 35

Scheduled kill

Number of Animals

Days from - to

 

10

91 91

 

10

91 91

 

10

91 91

 

10

91 91

Increased salivation

Number of Animals

Days from - to

 

.

.

 

.

.

 

.

.

 

10

5 90

Noisy respiration

Number of Animals

Days from - to

 

.

.

 

.

.

 

.

.

 

3

13 44

Table 6 Group Mean Body Weight Values

Body Weights (g)

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Day Numbers Relative to Start Date

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Group

(Sex)

 

1

8

15

22

29

36

43

50

57

64

71

78

85

91

1(M)

Mean

S.D.

N

201.8

6.6

10

240.9

5.6

10

270.9

6.6

10

295.5

8.8

10

314.2

10.6

10

328.2

13.1

10

343.6

16.1

10

352.8

16.9

10

365.9

18.7

10

372.7

18.3

10

385.8

18.8

10

392.8

20.6

10

400.2

22.6

10

405.9

22.7

10

2(M)

Mean

S.D.

N

200.9

10.5

10

240.4

15.0

10

273.1

22.3

10

299.3

27.6

10

319.6

32.2

10

335.5

34.9

10

350.0

36.4

10

361.2

37.1

10

372.4

41.0

10

381.2

42.8

10

391.2

42.7

10

398.6

42.8

10

404.9

 42.8

10

409.2

42.9

10

3(M)

Mean

S.D.

N

200.5

8.4

10

242.3

9.3

10

278.6

11.7

10

305.1

13.9

10

324.4

16.5

10

343.9

17.4

10

359.1

18.4

10

371.9

21.9

10

385.4

22.8

10

393

22.7

10

405.6

26.9

10

414.7

29.2

10

419.9

26.9

10

425

26.8

10

4(M)

Mean

S.D.

N

198.6

9.4

10

229.2

12.9

10

257.4

20.0

10

282

24.1

10

295.8

28.4

10

316.8

31.4

10

325.3

32.2

10

331.8

33.4

10

337.8

36.6

10

346.7

39.1

10

350.7

39.5

10

352.4

39.5

10

351

41.0

10

361.8

41.9

10

Table 6 Group Mean Body Weight Values

Body Weights (g)

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Day Numbers Relative to Start Date

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Group

(Sex)

 

1

8

15

22

29

36

43

50

57

64

71

78

85

91

1(F)

Mean

S.D.

N

148.3

6.3

10

167.1

9.2

10

183

8.7

10

189.7

8.6

10

198.9

13.9

10

206.2

13.0

10

213.9

13.0

10

218.4

12.3

10

223.1

13.1

10

224.7

13.8

10

231.4

12.8

10

234.2

12.3

10

235.9

14.7

10

238.4

14.4

10

2(F)

Mean

S.D.

N

145.4

4.6

10

169

3.7

10

182.4

5.6

10

192.8

6.4

10

200.9

3.5

10

211.6

8.2

10

218.3

8.9

10

221.7

7.3

10

226.2

5.2

10

232.4

8.9

10

236.6

8.5

10

237.7

7.4

10

241.8

6.1

10

243.8

8.3

10

3(F)

Mean

S.D.

N

148.8

7.5

10

170

10.0

10

180.9

11.8

10

188.5

13.2

10

201

13.3

10

210.5

15.5

10

216.7

18.7

10

219.6

19.8

10

225.9

18.0

10

231

18.6

10

232.3

19.5

10

234.4

18.8

10

240.5

18.3

10

239.8

17.8

10

4(F)

Mean

S.D.

N

147.1

6.8

10

165.4

5.5

10

180.1

8.7

10

188.2

10.4

10

195.4

12.4

10

206.4

10.2

10

211.9

12.3

10

215.2

11.9

10

216.8

12.8

10

219.3

13.4

10

223.6

14.1

10

226.6

14.0

10

227.1

14.6

10

231

14.3

10

Table 7 Group Mean Body Weight Gains

Body Weights (g)

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Day Numbers Relative to Start Date

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Group

(Sex)

From:

To:

1

8

8

15

15

22

22

29

29

36

36

43

43

50

50

57

57

64

64

71

71

78

78

85

85

91

1

91

1

91

1(M)

Mean

S.D.

N

39.1

4.1

10

30

6.3

10

24.6

3.5

10

18.7

3.4

10

14

4.2

10

15.4

5.8

10

9.2

3.3

10

13.1

4.9

10

6.8

3.4

10

13.1

3

10

7

3

10

7.4

4.4

10

5.7

2.9

10

204.1

27.5

10

101.6

16.5

10

2(M)

Mean

S.D.

N

39.5

6.2

10

32.7

8.2

10

26.2

6.2

10

20.3

5.8

10

15.9

4.1

10

14.5

4.2

10

11.2

4.4

10

11.2

5.2

10

8.8

3.8

10

10

4.3

10

7.4

3.4

10

6.3

3.6

10

4.3

3

10

208.3

37.4

10

103.6

17.3

10

3(M)

Mean

S.D.

N

41.8

4.5

10

36.3

6.3

10

26.5

4.1

10

19.3

5.4

10

19.5**

2.7

10

15.2

2.9

10

12.8

5.2

10

13.5

4

10

7.6

4.7

10

12.6

5.8

10

9.1

5.3

10

5.2

3.4

10

5.1

2.1

10

224.5

22

10

112

9.7

10

4(M)

Mean

S.D.

N

30.6**

7.3

10

28.2

9.7

10

24.6

7.2

10

13.8*

5.6

10

21.0**

5.6

10

8.5**

4.1

10

6.5

3.9

10

6.0**

5.6

10

8.9

6.5

10

4.0**

6.1

10

1.7*

6.1

10

-1.4**

6.6

10

10.8**

5.7

10

163.2

36.8

10

82

17.4

10

Table 7 Group Mean Body Weight Gains

Body Weights (g)

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Day Numbers Relative to Start Date

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Abs Gain

% Gain

Group

(Sex)

From:

To:

1

8

8

15

15

22

22

29

29

36

36

43

43

50

50

57

57

64

64

71

71

78

78

85

85

91

1

91

1

91

1(F)

Mean

S.D.

N

18.8

5.3

10

15.9

3.7

10

6.7

2.2

10

9.2

8.3

10

7.3

4.5

10

7.7

3.1

10

4.5

3.4

10

4.7

3.8

10

1.6

5.9

10

6.7

2.5

10

2.8

3.7

10

1.7

4.1

10

2.5

3.0

10

90.1

9.6

10

60.7

5.3

10

2(F)

Mean

S.D.

N

23.6

4.9

10

13.4

5.1

10

10.4

4.5

10

8.1

7.2

10

10.7

6.8

10

6.7

5.7

10

3.4

7.0

10

4.5

6.6

10

6.2

6.5

10

4.2

5.5

10

1.1

6.1

10

4.1

5.3

10

2.0

4.4

10

98.4

10.4

10

67.9

8.7

10

3(F)

Mean

S.D.

N

21.2

5.2

10

10.9

4.2

10

7.6

5.0

10

12.5

4.8

10

9.5

4.8

10

6.2

4.3

10

2.9

7.3

10

6.3

6.8

10

5.1

5.3

10

1.3*

3.1

10

2.1

7.9

10

6.1

6.1

10

-0.7

5.5

10

91.0

14.9

10

61.2

10.1

10

4(F)

Mean

S.D.

N

18.3

5.2

10

14.7

5.9

10

8.1

5.2

10

7.2

5.8

10

11.0

5.5

10

5.5

5.1

10

3.3

3.9

10

1.6

3.9

10

2.5

5.3

10

4.3*

2.6

10

3.0

3.6

10

0.5

3.3

10

3.9

3.2

10

83.9

10.1

10

57.0

6.2

10

Table 8 Group Mean Food Consumptions

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Day Numbers Relative to Start Date

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Group

(Sex)

From:

To:

1

8

8

15

15

22

22

29

29

36

36

43

43

50

50

57

57

64

64

71

71

78

78

85

85

91

1(M)

Mean

N

23.5

10

24.1

10

24.6

10

24.8

10

24.2

10

23.9

10

29.1

10

23.7

10

24

10

23.4

10

23.5

10

23.7

10

23

10

2(M)

Mean

N

23.3

10

24.9

10

24.9

10

25.6

10

24.8

10

24.2

10

24.9

10

24

10

24.8

10

23.7

10

23.4

10

24.4

10

22.9

10

3(M)

Mean

N

23.7

10

24.4

10

24.9

10

26.3

10

25.3

10

25.7

10

25.3

10

24.8

10

24.6

10

23.9

10

24.1

10

24.8

10

23.2

10

4(M)

Mean

N

21.5

10

23.3

10

24.2

10

25.3

10

24.2

10

23.9

10

22.6

10

21.7

10

22.5

10

21.4

10

22

10

20.6

10

21.2

10

Table 8 Group Mean Food Consumptions

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Day Numbers Relative to Start Date

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Group

(Sex)

From:

To:

1

8

8

15

15

22

22

29

29

36

36

43

43

50

50

57

57

64

64

71

71

78

78

85

85

91

1(F)

Mean

N

15.9

10

17.4

10

16.3

10

17.8

10

18.3

10

17.2

10

17.3

10

17.9

10

17.5

10

16.9

10

17.5

10

16.3

10

17

10

2(F)

Mean

N

16.4

10

17.9

10

16.6

10

18.8

10

19.1

10

18.1

10

18.1

10

19

10

18.3

10

18.5

10

18

10

17.6

10

17.6

10

3(F)

Mean

N

16.4

10

17.3

10

16.2

10

18.4

10

18.4

10

17.7

10

16.5

10

18.8

10

17.9

10

17.6

10

17.8

10

16.8

10

16.6

10

4(F)

Mean

N

15.7

10

17.3

10

16.1

10

17.6

10

17.9

10

16.6

10

17.1

10

16.7

10

17.2

10

16.2

10

17

10

15.8

10

16.1

10

Table 9 Food Efficiency

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Day Numbers Relative to Start Date

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Group

(Sex)

From:

To:

1

8

8

15

15

22

22

29

29

36

36

43

43

50

50

57

57

64

64

71

71

78

78

85

85

91

1(M)

Mean

N

23.8

17.8

14.3

10.7

8.2

9.2

4.7

7.9

4.1

8

4.2

4.4

4.2

10

10

10

10

10

10

10

10

10

10

10

10

10

2(M)

Mean

N

24.2

18.7

15

11.2

9.2

8.5

6.4

6.6

5.1

6

4.5

3.7

3.1

10

10

10

10

10

10

10

10

10

10

10

10

10

3(M)

Mean

N

25.2

21.2

15.3

10.5

11

8.5

7.2

7.8

4.4

7.4

5.3

3.1

3.7

10

10

10

10

10

10

10

10

10

10

10

10

10

4(M)

Mean

N

20.3

17.3

14.5

7.8

12.4

5

4.2

4

5.6

2.6

1.1

-1.0

8.5

10

10

10

10

10

10

10

10

10

10

10

10

10

Table 9 Food Efficiency

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Day Numbers Relative to Start Date

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Group

(Sex)

From:

To:

1

8

8

15

15

22

22

29

29

36

36

43

43

50

50

57

57

64

64

71

71

78

78

85

85

91

1(F)

Mean

N

16.8

13.1

6

7.1

5.8

6.5

3.9

3.8

1.4

5.7

2.4

1.4

2.5

10

10

10

10

10

10

10

10

10

10

10

10

10

2(F)

Mean

N

20.6

10.7

8.9

6.2

8

5.3

2.7

3.4

4.8

3.3

0.9

3.3

1.9

10

10

10

10

10

10

10

10

10

10

10

10

10

3(F)

Mean

N

18.5

9

6.7

9.7

7.2

4.9

2.8

4.8

3.9

1

1.8

5.2

ƒ0.7

10

10

10

10

10

10

10

10

10

10

10

10

10

4(F)

Mean

N

16.6

12.2

7.1

5.9

8.8

4.6

2.9

1.3

2

3.8

2.6

0.5

4.1

10

10

10

10

10

10

10

10

10

10

10

10

10

Table 10 Group Mean Hematological Values

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Group

(Sex)

 

Hb

g/dl

RBC

10^12/l

Hct

%

MCH

pg

MCV

fl

MCHC

g/dl

WBC

10^9/l

Neut

10^9/l

Lymph

10^9/l

Mono

10^9/l

Eos

10^9/l

Bas

10^9/l

CT

Seconds

PLT

10^9/l

APTT

Seconds

1(M)

Mean

17.07

9.633

50.67

17.72

52.63

33.67

7.02

1.128

5.837

0.000n

0.057

0.000n

9.16

641.8

14.02

SD

0.78

0.484

2.43

0.53

1.4

0.29

1.2

0.297

1.023

0

0.054

0

0.63

60.4

1.04

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

2(M)

Mean

16.69

9.762

49.85

17.13

51.1

33.47

6.67

1.327

5.279

0.014n

0.052

0.000n

9.42

686.6

13.55

SD

0.55

0.389

1.41

0.58

1.43

0.4

1.18

0.352

1.081

0.03

0.088

0

0.43

106.7

1.12

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

3(M)

Mean

16.67

9.257

49.7

18.01

53.7

33.54

7.06

1.583*

5.429

0.014n

0.036

0.000n

9.48

660.6

14.15

SD

0.54

0.263

1.74

0.55

1.59

0.37

1.29

0.398

1.136

0.03

0.054

0

0.39

63.3

1.22

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

4(M)

Mean

14.87**

8.732**

44.15**

17.06*

50.62**

33.7

7.38

1.494*

5.822

0.000n

0.064

0.000n

9.33

620.9

13.5

SD

0.93

0.709

2.86

0.59

1.3

0.44

1.3

0.550

1.012

0

0.08

0

0.4

103.9

1.09

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

Table 10 Group Mean Hematological Values

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Group

(Sex)

 

Hb

g/dl

RBC

10^12/l

Hct

%

MCH

pg

MCV

fl

MCHC

g/dl

WBC

10^9/l

Neut

10^9/l

Lymph

10^9/l

Mono

10^9/l

Eos

10^9/l

Bas

10^9/l

CT

Seconds

PLT

10^9/l

APTT

Seconds

1(F)

Mean

15.38

8.062

45.11

19.1

55.95

34.15

4.2

0.865

3.3

0.004n

0.033

0.000n

9.43

618.7

14.21

SD

1.48

0.832

4.66

0.5

1.45

0.31

1.37

0.504

0.914

0.013

0.039

0

0.63

55.7

1.64

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

2(F)

Mean

15.9

8.345

47.1

19.06

56.49

33.75

5.04

1.138

3.877

0.005n

0.022

0.000n

9.51

583.6

15.17

SD

0.59

0.381

2.11

0.43

1.2

0.49

1.51

0.551

1.075

0.016

0.037

0

0.66

195.6

1.49

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

3(F)

Mean

15.72

8.186

46.33

19.19

56.61

33.94

4.49

0.929

3.55

0.000n

0.012

0.000n

9.16

649.1

13.8

SD

0.9

0.424

2.85

0.5

1.59

0.23

1.12

0.354

0.929

0

0.027

0

0.56

55.4

1.29

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

4(F)

Mean

14.91

8.105

44.01

18.39**

54.34*

33.86

5.91**

1.241

4.647**

0.000n

0.023

0.000n

9.02

567

14.77

SD

0.83

0.455

2.26

0.48

1.3

0.24

1.16

0.229

1.144

0

0.039

0

0.61

171.8

2.04

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

Table 11 Group Mean Blood Chemical Values

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Group

(Sex)

 

Urea

mg/dl

Glucose

mg/dl

Tot.

Prot.

g/dl

Albumin

g/dl

A/G

Ratio

Na+

mmol/l

K+

mmol/l

Clƒ

mmol/l

Ca++

mmol/l

P

mmol/l

ASAT

IU/l

ALAT

IU/l

AP

IU/l

Creat

mg/dl

Chol

mg/dl

Bili

mg/dl

Bile Acid

μmol/

1(M)

Mean

34.6

154.5

6.957

3.84

1.221

149.2

4.381

103.2

2.658

2.01

86.3

70.2

136.1

0.649

111.4

0.134

35.99

SD

2.6

13.5

0.348

0.18

0.066

2.6

0.465

1.7

0.103

0.38

29.2

33.4

23.8

0.046

17.8

0.027

32.45

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

2(M)

Mean

39.1*

170.9*

6.717

3.7

1.232

147.4

4.25

103.1

2.606

1.8

81.2

58.9

127.9

0.661

100.6

0.121

31.6

SD

3.7

14.9

0.303

0.13

0.054

2.3

0.22

1

0.089

0.33

17.2

7.1

27.8

0.078

11.3

0.026

18.65

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

3(M)

Mean

36.0*

154

6.893

3.74

1.185

148.1

7.615

102.6

2.369

1.85

89

64.3

115.4

0.649

131.4**

0.153

47.32

SD

3.1

14.8

0.289

0.12

0.075

3.3

10.55

1.9

0.836

0.26

18.8

22.3

47.3

0.046

14.5

0.027

25.33

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

4(M)

Mean

39.1**

161.5

6.228**

3.54**

1.327**

149

4.275

102

2.871*

2.32*

116.7*

77.7

109.6

0.658

137.2**

0.227**

109.74**

SD

4.1

15.3

0.602

0.33

0.067

2.6

0.406

1.3

0.179

0.31

34.9

15.7

17.1

0.077

20.8

0.05

40.63

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

Table 11 Group Mean Blood Chemical Values

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Group

(Sex)

 

Urea

mg/dl

Glucose

mg/dl

Tot.

Prot.

g/dl

Albumin

g/dl

A/G

Ratio

Na+

mmol/l

K+

mmol/l

Clƒ

mmol/l

Ca++

mmol/l

P

mmol/l

ASAT

IU/l

ALAT

IU/l

AP

IU/l

Creat

mg/dl

Chol

mg/dl

Bili

mg/dl

Bile Acid

μmol/

1(F)

Mean

49.9

148

6.959

4.11

1.46

148.8

5.116

102.5

3.013

1.92

98.9

63.6

79.7

0.73

98.9

0.108

64.46

SD

14.2

13.7

0.791

0.41

0.116

2.4

1.356

2

0.321

0.41

33

13.5

28.2

0.197

24.4

0.028

20.53

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

2(F)

Mean

50.9

156.2

6.992

4.05

1.404

148.4

5.071

103.4

3.035

1.74

90.2

59.9

67

0.771

98.9

0.103

41.1

SD

23.2

20.9

0.607

0.3

0.134

2.6

2.125

2.4

0.146

0.65

28.6

11.6

19.4

0.368

17

0.024

27.88

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

3(F)

Mean

41.8

145.2

7.434

4.36

1.421

149.6

4.345

103.4

3.145

1.98

80.6

57.9

60.6

0.7

104

0.146**

95.55

SD

6.1

11.7

0.418

0.22

0.082

2.6

0.313

3.2

0.093

0.34

17.3

15.2

15.9

0.081

14.7

0.02

63.3

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

4(F)

Mean

46.6

145.3

6.042**

3.59**

1.47

150.3

6.472

102.6

3.113

2.02

125.6

82.1*

65.9

0.656

116.1

0.174**

110.61

SD

4.7

14.9

0.883

0.54

0.129

6.5

6.47

3.7

0.606

0.26

68

25

20.8

0.109

26.3

0.082

67.07

N

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

Table 2 Summary Incidence of Behavioral Assessments (Observations)

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Day Numbers Relative to Start Date

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Group

(Sex)

Clinical Sign

-1

6

13

20

27

34

41

48

55

62

69

76

82

88

1(M)

ANIMALS ALIVE

10

10

10

10

10

10

10

10

10

10

10

10

10

10

ANIMALS NORMAL

10

10

10

10

10

10

10

10

10

10

10

10

10

10

1(F)

ANIMALS ALIVE

10

10

10

10

10

10

10

10

10

10

10

10

10

10

ANIMALS NORMAL

10

10

10

10

10

10

10

10

10

10

10

10

10

10

2(M)

ANIMALS ALIVE

10

10

10

10

10

10

10

10

10

10

10

10

10

10

ANIMALS NORMAL

10

10

10

10

10

10

10

10

10

10

10

10

10

10

2(F)

ANIMALS ALIVE

10

10

10

10

10

10

10

10

10

10

10

10

10

10

ANIMALS NORMAL

10

10

10

10

10

10

10

10

10

10

10

10

10

10

3(M)

ANIMALS ALIVE

10

10

10

10

10

10

10

10

10

10

10

10

10

10

ANIMALS NORMAL

10

10

10

10

10

10

10

10

10

10

10

10

10

10

3(F)

ANIMALS ALIVE

10

10

10

10

10

10

10

10

10

10

10

10

10

10

ANIMALS NORMAL

10

10

10

10

10

10

10

10

10

10

10

10

10

10

4(M)

ANIMALS ALIVE

10

10

10

10

10

10

10

10

10

10

10

10

10

10

ANIMALS NORMAL

10

10

10

10

10

10

10

10

10

10

10

10

10

10

4(F)

ANIMALS ALIVE

10

10

10

10

10

10

10

10

10

10

10

10

10

10

ANIMALS NORMAL

10

10

10

10

10

10

10

10

10

10

10

10

10

10

Table 3 Group Mean Behavioral Assessments (Scores)

 

 

 

 

 

Day: -1 Relative to Start Date

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(M)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

3(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(M)

Mean

0.0

0.1

0.1

0.0

SD

0.0

0.3

0.3

0.0

N

10

10

10

10

Table 3 Group Mean Behavioral Assessments (Scores)

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Day: 6 Relative to Start Date

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(M)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

3(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Table 3 Group Mean Behavioral Assessments (Scores)

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Day: 13 Relative to Start Date

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Table 3 Group Mean Behavioral Assessments (Scores)

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Day: 20 Relative to Start Date

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(M)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

4(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Table 3 Group Mean Behavioral Assessments (Scores)

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(F)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

Day: 27 Relative to Start Date

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(M)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

Table 3 Group Mean Behavioral Assessments (Scores)

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(F)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

2(F)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

3(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(F)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

Day: 34 Relative to Start Date

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Table 3 Group Mean Behavioral Assessments (Scores)

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(F)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

2(F)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

3(F)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

4(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Day: 41 Relative to Start Date

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(M)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

4(M)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

Table 3 Group Mean Behavioral Assessments (Scores)

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(F)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

3(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(F)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

Day: 48 Relative to Start Date

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Table 3 Group Mean Behavioral Assessments (Scores)

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Day: 55 Relative to Start Date

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(M)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

2(M)

Mean

0.0

0.2

0.1

0.0

SD

0.0

0.4

0.3

0.0

N

10

10

10

10

3(M)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

4(M)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

Table 3 Group Mean Behavioral Assessments (Scores)

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Day: 62 Relative to Start Date

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(M)

Mean

0.0

0.1

0.0

0.0

SD

0.0

0.3

0.0

0.0

N

10

10

10

10

2(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Table 3 Group Mean Behavioral Assessments (Scores)

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Day: 69 Relative to Start Date

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Table 3 Group Mean Behavioral Assessments (Scores)

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Day: 76 Relative to Start Date

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Table 3 Group Mean Behavioral Assessments (Scores)

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Day: 82 Relative to Start Date

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(M)

Mean

0.0

0.1

0.1

0.0

SD

0.0

0.3

0.3

0.0

N

10

10

10

10

2(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(M)

Mean

0.0

0.0

0.1

0.0

SD

0.0

0.0

0.3

0.0

N

10

10

10

10

4(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Table 3 Group Mean Behavioral Assessments (Scores)

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(F)

Mean

0.2

0.0

0.0

0.0

SD

0.6

0.0

0.0

0.0

N

10

10

10

10

4(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Day: 88 Relative to Start Date

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

2(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(M)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

Table 3 Group Mean Behavioral Assessments (Scores)

 

 

 

 

 

Group

(Sex)

 

Transfer arousal

Urination

Defecation

Tail elevation

1(F)

Mean

0.0

0.0

0.1

0.0

SD

0.0

0.0

0.3

0.0

N

10

10

10

10

2(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

3(F)

Mean

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

N

10

10

10

10

4(F)

Mean

0.0

0.2

0.0

0.0

SD

0.0

0.4

0.0

0.0

N

10

10

10

10

Table 4 Group Mean Functional Performance Test Values

General Footnote: Unit = Time (seconds) for Motor Activity Assessments

 

 

 

 

 

 

 

 

 

 

Group

(Sex)

 

Test 1

Forelimb

g

Test 1

Hindlimb

g

Test 2

Forelimb

g

Test 2

Hindlimb

g

Test 3

Forelimb

g

Test 3

Hindlimb

g

Overall

activity

Overall

mobile

Last 20%

activity

Last 20%

mobile

1(M)

Mean

1186

505

1363.5

583.1

1332.9

477.8

604.9

0.0n

57.8

0.0n

SD

303.1

143.9

245.2

141

190.3

200.9

146.7

0

72

0

N

10

10

10

10

10

10

10

10

10

10

2(M)

Mean

1214.2

545.5

1391.5

491.3

1278.3

476.9

412.1*

0.0n

21.2

0.0n

SD

215.1

153.5

138.7

162.2

182.5

126.6

119.9

0

21.6

0

N

10

10

10

10

10

10

10

10

10

10

3(M)

Mean

1084

423.4

1236

445.3

1270.1

512.2

455.8*

0.0n

49.8

0.0n

SD

226.9

199.6

276.7

175.5

320.8

89.6

145.7

0

67.2

0

N

10

10

10

10

10

10

10

10

10

10

4(M)

Mean

1066.5

517.9

1219

506.1

1279.5

492.1

462.2*

0.0n

13

0.0n

SD

274.7

120.8

261.2

122.5

260.2

161.6

129.6

0

13.4

0

N

10

10

10

10

10

10

10

10

10

10

Table 4 Group Mean Functional Performance Test Values

 

 

 

 

 

 

 

 

 

 

 

Group

(Sex)

 

Test 1

Forelimb

g

Test 1

Hindlimb

g

Test 2

Forelimb

g

Test 2

Hindlimb

g

Test 3

Forelimb

g

Test 3

Hindlimb

g

Overall

activity

Overall

mobile

Last 20%

activity

Last 20%

mobile

1(F)

Mean

876.4

351.7

1081.3

373.9

1148.9

366.4

585.6

0.0n

71.1

0.0n

SD

190.9

87.1

153.4

54.2

182.1

120.6

214.5

0

70.1

0

N

10

10

10

10

10

10

10

10

10

10

2(F)

Mean

1117

402.7

1131.9

366.4

1125.2

356.3

645.2

0.0n

43

0.0n

SD

312.2

166.4

124.1

83.9

202.4

80.6

183.2

0

50.1

0

N

10

10

10

10

10

10

10

10

10

10

3(F)

Mean

969.8

396.4

1155.5

435.9

1115.7

372.4

569.7

0.0n

59.6

0.0n

SD

314.9

122.1

321.3

124.6

279.2

177.5

142.7

0

57.5

0

N

10

10

10

10

10

10

10

10

10

10

4(F)

Mean

972.4

381.3

1025.6

400.9

1056.8

371.6

486

0.0n

61.3

0.0n

SD

149

161.3

247.3

139.2

202.8

98.7

131.2

0

67.9

0

N

10

10

10

10

10

10

10

10

10

10

Table 5 Group Mean Sensory Reactivity Assessments (Scores)

General Footnote: Unit = Time (seconds) for Motor Activity Assessments

 

 

 

 

 

 

 

 

 

Group

(Sex)

 

Grasp

response

Vocalisation

Toe

pinch

Tail

pinch

Finger

approach

Touch

escape

Pupil

reflex

Blink

reflex

Startle

reflex

1(M)

Mean

0.0

0.1

0.0

0.0

0.0

0.0

0.0

0.0

0.0

SD

0.0

0.3

0.0

0.0

0.0

0.0

0.0

0.0

0.0

N

10

10

10

10

10

10

10

10

10

2(M)

Mean

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

N

10

10

10

10

10

10

10

10

10

3(M)

Mean

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

N

10

10

10

10

10

10

10

10

10

4(M)

Mean

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

N

10

10

10

10

10

10

10

10

10

Table 5 Group Mean Sensory Reactivity Assessments (Scores)

 

 

 

 

 

 

 

 

 

 

Group

(Sex)

 

Grasp

response

Vocalisation

Toe

pinch

Tail

pinch

Finger

approach

Touch

escape

Pupil

reflex

Blink

reflex

Startle

reflex

1(F)

Mean

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

N

10

10

10

10

10

10

10

10

10

2(F)

Mean

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

N

10

10

10

10

10

10

10

10

10

3(F)

Mean

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

N

10

10

10

10

10

10

10

10

10

4(F)

Mean

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

SD

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

0.0

N

10

10

10

10

10

10

10

10

10

Table 13 Group Mean Organ Weights with Corresponding Relative (% of Body Weight) Organ Weights)

 

males

 

 

 

females

 

 

 

Group

(Sex)

 

0

Control

10

mg/kg bw/day

100

mg/kg bw/day

1000

mg/kg bw/day

0

Control

10

mg/kg bw/day

100

mg/kg bw/day

100

mg/kg bw/day

Terminal Bodyweight

Mean (g)

405.9

409.2

425

361.8

238.4

243.8

239.8

231

SD

22.7

42.9

26.8

41.9

14.4

8.3

17.8

14.3

N

10

10

10

10

10

10

10

10

Adrenals

Mean (g)

0.14254

0.0823

0.07924

0.08482

0.09092

0.08768

0.09016

0.094

SD

0.19824

0.01195

0.01061

0.02121

0.02087

0.0128

0.01002

0.01121

N

10

10

10

10

10

10

10

10

 

Mean (%)

0.035

0.02

0.019

0.024

0.038

0.036

0.038

0.041

SD

0.049

0.003

0.003

0.008

0.008

0.005

0.005

0.006

N

10

10

10

10

10

10

10

10

Brain (Including Cerebrum,

Cerebellum And Pons)

Mean (g)

2.03786

2.02169

2.01967

1.99041

1.88639

1.89455

1.88843

1.9018

SD

0.08495

0.0778

0.09075

0.11528

0.08272

0.09452

0.07901

0.08255

N

10

10

10

10

10

10

10

10

 

Mean (%)

0.504

0.498

0.477

0.555

0.793

0.777

0.791

0.825

SD

0.038

0.05

0.031

0.057

0.041

0.032

0.059

0.044

N

10

10

10

10

10

10

10

10

Epididymides

Mean (g)

1.67461

1.74198

1.64516

1.4918

 

 

 

 

SD

0.08524

0.22788

0.09166

0.15543

 

 

 

 

N

10

10

10

10

 

 

 

 

 

Mean (%)

0.414

0.429

0.388

0.415

 

 

 

 

SD

0.031

0.061

0.029

0.047

 

 

 

 

N

10

10

10

10

 

 

 

 

Heart

Mean (g)

1.29981

1.28729

1.36371

1.10857

0.83674

0.88342

0.88545

0.78914

SD

0.19976

0.20968

0.22931

0.16105

0.08941

0.094

0.09108

0.08512

N

10

10

10

10

10

10

10

10

 

Mean (%)

0.322

0.314

0.321

0.306

0.351

0.363

0.369

0.342

SD

0.057

0.039

0.053

0.025

0.03

0.039

0.031

0.031

N

10

10

10

10

10

10

10

10

Kidneys

Mean (g)

2.5076

2.4324

2.49301

2.84881**

1.62093

1.62368

1.49202

1.6022

SD

0.2686

0.30265

0.27463

0.38253

0.12945

0.13374

0.36762

0.11979

N

10

10

10

10

10

10

10

10

 

Mean (%)

0.62

0.594

0.587

0.791**

0.681

0.666

0.623

0.694

SD

0.079

0.031

0.061

0.088

0.05

0.048

0.144

0.04

N

10

10

10

10

10

10

10

10

Liver

Mean (g)

14.0973

13.5228

13.5072

12.7207

8.37531

8.61163

8.62506

8.89275**

SD

0.80738

1.4142

1.39253

1.74243

0.72176

0.48659

1.41325

0.84432

N

10

10

10

10

10

10

10

10

 

Mean (%)

3.477

3.313

3.18

3.516

3.512

3.533

3.585

3.849**

SD

0.192

0.241

0.29

0.214

0.191

0.168

0.395

0.254

N

10

10

10

10

10

10

10

10

Ovaries

Mean (g)

 

 

 

 

0.11128

0.11889

0.10826

0.10047

SD

 

 

 

 

0.02625

0.01757

0.02406

0.01776

N

10

10

10

10

10

10

10

10

 

Mean (%)

 

 

 

 

0.047

0.049

0.045

0.044

SD

 

 

 

 

0.01

0.008

0.01

0.008

N

10

10

10

10

10

10

10

10

Spleen

Mean (g)

0.83122

0.78951

0.72391

0.77071

0.54942

0.56923

0.55243

0.52266

SD

0.14367

0.1681

0.09731

0.06243

0.06112

0.04592

0.06587

0.05779

N

10

10

10

10

10

10

10

10

 

Mean (%)

0.205

0.193

0.171

0.215

0.231

0.233

0.231

0.226

SD

0.034

0.037

0.025

0.023

0.022

0.015

0.023

0.02

N

10

10

10

10

10

10

10

10

Testes

Mean (g)

3.9234

3.89172

3.80977

3.67888

 

 

 

 

SD

0.25766

0.2624

0.35554

0.31946

 

 

 

 

N

10

10

10

10

 

 

 

 

 

Mean (%)

0.97

0.958

0.898

1.03

 

 

 

 

SD

0.084

0.093

0.084

0.157

 

 

 

 

N

10

10

10

10

 

 

 

 

Thymus

Mean (g)

0.54632

0.55843

0.55512

0.44133

0.39568

0.44845

0.38762

0.39191

SD

0.08738

0.08574

0.08877

0.11014

0.07312

0.05263

0.08178

0.10918

N

10

10

10

10

10

10

10

10

 

Mean (%)

0.134

0.137

0.131

0.122

0.168

0.184

0.162

0.168

SD

0.019

0.017

0.024

0.026

0.039

0.021

0.031

0.041

N

10

10

10

10

10

10

10

10

Uterus (With Cervix)

Mean (g)

 

 

 

 

0.85005

0.75992

0.76975

0.88066

SD

 

 

 

 

0.26502

0.21015

0.31299

0.29646

N

10

10

10

10

10

10

10

10

 

Mean (%)

 

 

 

 

0.359

0.313

0.32

0.381

SD

 

 

 

 

0.115

0.091

0.121

0.122

N

10

10

10

10

10

10

10

10

Table 12 Summary Incidence of Necropsy Findings

Males

 

 

 

Removal Reason: TERMINAL SACRIFICE

 

Number of Animals Examined :

0

Control

10

10

mg/kg bw/day

10

100

mg/kg bw/day

10

1000

mg/kg bw/day

10

Kidneys;

Submitted

(10)

(10)

(10)

(10)

No Visible Lesions

10

10

9

4

Discolouration; pale

0

0

0

5

Enlarged

0

0

0

4

Increased Pelvic Space; left

0

0

1

0

Lungs (With Bronchi);

Submitted

(10)

(10)

(10)

(10)

No Visible Lesions

9

9

9

9

Discolouration; red

1

1

1

1

Stomach;

Submitted

(10)

(10)

(10)

(10)

No Visible Lesions

9

10

10

9

Raised; limiting ridge

1

0

0

0

Thickened; glandular region

0

0

0

1

 

Females

 

 

 

Removal Reason: TERMINAL SACRIFICE

 

Number of Animals Examined :

0

Control

10

10

mg/kg bw/day

10

100

mg/kg bw/day

10

1000

mg/kg bw/day

10

Kidneys;

Submitted

(10)

(10)

(10)

(10)

No Visible Lesions

9

10

10

10

Increased Pelvic Space; left

1

0

0

0

Lungs (With Bronchi);

Submitted

(10)

(10)

(10)

(10)

No Visible Lesions

8

8

8

7

Discolouration

1

0

0

1

Discolouration; red

1

2

2

2

Applicant's summary and conclusion

Conclusions:
The oral administration of 4,4’-Isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane, reaction products with fatty acids, C18-unsatd., dimers, ERRCF #62 to rats for a period of ninety consecutive days, at dose levels of 10, 100 or 1000 mg/kg bw/day resulted in treatment related effects in animals of either sex treated with 1000 mg/kg bw/day. The No Observed Effect Level (NOEL) for animals of either sex was therefore considered to be 100 mg/kg bw/day.
Executive summary:

Introduction

The study was designed to investigate the systemic toxicity of the test item and is compatible with the following regulatory guideline:

• The OECD Guidelines for Testing of Chemicals No. 408 "Subchronic Oral Toxicity - Rodent: 90 Day Study” (Adopted 21 September 1998).

This study was also designed to be compatible with Commission Regulation (EC) No 440/2008 of 30 May 2008, laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH).

Methods

The test item was administered by gavage to three groups, each of ten male and ten female Wistar Han™:RccHan™:WIST strain rats, for ninety consecutive days, at dose levels of 10, 100 and 1000 mg/kg bw/day. A control group of ten males and ten females was dosed with vehicle alone (Polyethylene glycol 400).

Clinical signs, functional observations, body weight change, dietary intake and water consumption were monitored during the study. Hematology and blood chemistry were evaluated for all animals at the end of the study. Ophthalmoscopic examination was also performed on control group and high dose animals.

All animals were subjected to gross necropsy examination and histopathological evaluation of selected tissues was performed.

Results

Mortality

There were no unscheduled deaths.

Clinical Observations

Increased salivation and noisy respiration was evident in animals of either sex treated with 1000 mg/kg bw/day during the treatment period. An isolated incident of increased salivation was evident in one male treated with 100 mg/kg bw/day. Noisy respiration was also evident in one male treated with 100 mg/kg bw/day on two days and an isolated incident was evident in one male treated with 10 mg/kg bw/day. No such effects were detected in females treated with 100 or 10 mg/kg bw/day.

Behavioral Assessment

There were no treatment-related changes in the behavioral parameters at 10, 100 or 1000 mg/kg bw/day.

Functional Performance Tests

There were no treatment-related changes in functional performance considered to be related to treatment at 10, 100 or 1000 mg/kg bw/day.

Sensory Reactivity Assessments

There were no inter-group differences in sensory reactivity scores that were considered to be related to treatment at 10, 100 or 1000 mg/kg bw/day.

Body Weight

Males treated with 1000 mg/kg bw/day showed a reduction in overall body weight gain. Females from this treatment group also showed a slight reduction in overall body weight gain. No such effects were evident in animals of either sex treated with 100 or 10 mg/kg bw/day.

Food Consumption

Males treated with 1000 mg/kg bw/day showed a slight reduction in overall food consumption and reductions in food conversion efficiency. No such effects were evident in females treated with 1000 mg/kg bw/day or in animals of either sex treated with 100 or 10 mg/kg bw/day.

Water Consumption

No adverse effect on water consumption was evident in treated animals.

Ophthalmoscopy

Ophthalmic examination of the eyes from rats treated with 1000 mg/kg bw/day did not indicate any effect of treatment.

Hematology

Males treated with 1000 mg/kg bw/day showed reductions in hemoglobin, erythrocyte count, hematocrit, mean corpuscular hemoglobin and mean corpuscular volume. Females treated with 1000 mg/kg bw/day showed increases in total leucocyte count and lymphocytes. No toxicologically significant effects were detected in animals of either sex treated with 100 or 10 mg/kg bw/day.

Blood Chemistry

Animals of either sex treated with 1000 mg/kg bw/day showed reductions in total protein and albumin and an increase in bilirubin. Males treated with 1000 mg/kg bw/day also showed increases in albumin/globulin ratio, aspartate aminotransferase, cholesterol, bile acids, calcium concentration and phosphorus whilst females from this treatment group also showed increases in alanine aminotransferase and bile acids. No toxicologically significant effects were detected in animals of either sex treated with 100 or 10 mg/kg bw/day.

Necropsy

Five males treated with 1000 mg/kg bw/day had pale kidneys. Two of these males also had enlarged kidneys and one of these males had enlarged kidneys and a thickened glandular region of the stomach. Another male from this treatment group also had enlarged kidneys without associated pallor. No toxicologically significant effects were evident in females treated with 1000 mg/kg bw/day or in animals of either sex treated with 100 or 10 mg/kg bw/day.

Organ Weights

Females treated with 1000 mg/kg bw/day showed an increase in liver weight both absolute and relative to terminal body weight. Males treated with 1000 mg/kg bw/day showed an increase in kidney weights both absolute and relative to terminal body weight. No treatment related effects were detected in animals of either sex treated with 100 or 10 mg/kg bw/day.

Histopathology

The following treatment related microscopic abnormalities were detected:

Caecum: Mucosal hypertrophy was evident in eight males treated with 1000 mg/kg bw/day. No such effects were evident in females treated with 1000 mg/kg bw/day or in animals of either sex treated with 100 or 10 mg/kg bw/day.

Small Intestine: Lymphangiectasis was evident in the villi of the jejunum or ileum in five males and six females treated with 1000 mg/kg bw/day. No such effects were evident in animals of either sex treated with 100 or 10 mg/kg bw/day.

Kidneys: Multifocal basophilic tubules were evident in eight males and four females treated with 1000 mg/kg bw/day. Tubular dilation was evident in four males and five females from this treatment group and mineralization in the pelvis of the kidney was also evident in five males and females. No such effects were evident in animals of either sex treated with 100 or 10 mg/kg bw/day.

Liver: Minimal or mild centrilobular hypertrophy was evident in five males and seven females treated with 1000 mg/kg bw/day. No such effects were evident in animals of either sex treated with 100 or 10 mg/kg bw/day.

Ovaries: Interstitial cell hyperplasia was evident in nine females treated with 1000 mg/kg bw/day. No such effects were evident in females treated with 100 or 10 mg/kg bw/day.

Conclusion

The oral administration of 4,4’-Isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane, reaction products with fatty acids, C18-unsatd., dimers, ERRCF #62 to rats for a period of ninety consecutive days, at dose levels of 10, 100 or 1000 mg/kg bw/day resulted in treatment related effects in animals of either sex treated with 1000 mg/kg bw/day. The No Observed Effect Level (NOEL) for animals of either sex was therefore considered to be 100 mg/kg bw/day.