Registration Dossier

Diss Factsheets

Toxicological information

Repeated dose toxicity: oral

Currently viewing:

Administrative data

Endpoint:
repeated dose toxicity: oral, other
Remarks:
Combined Repeated Dose Toxicity Study with the Reproduction/Developmental Toxicity Screening Test by Oral Gavage in Sprague Dawley Rats
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15 June 2021 to 22 December 2021
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-reference
Reason / purpose for cross-reference:
reference to same study
Reference
Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
key study
Study period:
15 June 2021 to 22 December 2021
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
As per OECD Guideline for Testing of Chemicals, Number 422, adopted on 29 July 2016
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Justification for study design:
- Basis for dose level selection: There was no information or data available from repeated dose/reproduction toxicity and teratology studies for the test item.
The doses of 0, 111, 333 and 1000 mg/kg body weight/day were selected as control, low, mid and high dose groups for the dose range finding study (BIO-DTX 056) in consultation with the sponsor.
There were no test item-related related changes noted in any of the systemic, reproduction and developmental parameters up to the dose level of 1000 mg/kg body weight/day in the Dose Range Finding Study (BIO-DTX 056).
Hence, the same doses of 0, 111, 333 and 1000 mg/kg body weight/day were selected as control, low, mid and high dose groups respectively, for the present study in consultation with the sponsor.
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: 21 to 29ºC
- Stability under test conditions: The test item was stable for 6 hours and 48 hours at room temperature in 0.5% w/v Carboxymethyl cellulose with the concentrations of 5 mg/mL and 100 mg/mL.
- Solubility of the test substance: The test item was insoluble in distilled water and uniformly suspended in 0.5% w/v Carboxy Methyl Cellulose at the concentration of 100 mg/mL (the highest dose concentration selected for the study considering the dose volume of 10 mL/kg body weight) as per in-house solubility/suspendibility test results.
Hence, 0.5% w/v Carboxymethyl Cellulose was used as vehicle for test item formulations in consultation with sponsor.
0.5% w/v Carboxymethyl Cellulose is a routinely used and universally accepted vehicle of choice for the oral toxicity studies.

Species:
rat
Strain:
Sprague-Dawley
Details on species / strain selection:
Rat is one of the standard laboratory rodent species used for toxicity assessment and also recommended by various regulatory authorities.
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Hylasco Biotechnology India Pvt. Ltd, Charles River Technology Licensee,
CPCSEA (Committee for the Purpose of Control and Supervision of Experiments on Animals) Registration No.: 1808/PO/RcBt/S/15/CPCSEA.
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 8 to 10 weeks
- Weight at study initiation: Males: 254.36 to 291.64 g; Females: 200.54 to 211.92 g
- Housing: Animals were housed in a standard polypropylene cage (size: L 430 x B 285 x H 150 mm) with stainless steel mesh top grill having facilities for holding pelleted food and drinking water in water bottle fitted with stainless steel sipper tube. Clean sterilized paddy husk was provided as bedding material.
During acclimatization, two animals of same sex were housed.
i. Main Group Animals:
- Pre mating: Per cage two animals of same sex and group were housed.
- Cohabitation Period (mating): Per cage two animals (one male and one female) of same group were housed.
- Post-mating: After confirmation on presence of sperm in the vaginal smear (Day 0 of pregnancy), the mated pairs were separated. Males were housed with their former cage mates while females were housed individually. Sterilized paper shreds were provided as a nesting material for main group females from gestation day 20 onwards.
ii. Recovery Group Animals:
- Maximum of two animals per cage per group of same sex were housed, tunnels were provided as enrichment for the animals which were housed individually.
- Diet (e.g. ad libitum): Altromin maintenance diet
- Water (e.g. ad libitum): Water was provided ad libitum throughout the experimental period. Deep bore-well water passed through reverse osmosis unit was provided in plastic water bottles with stainless steel sipper tubes.
- Acclimation period: 5 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.9 to 22.9 oC
- Humidity (%): 48 to 63%
- Air changes (per hr): 12 to 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hours fluorescent light and 12 hours dark cycle
IN-LIFE DATES: From: 24 June 2021 To: 16 September 2021
Route of administration:
oral: gavage
Details on mating procedure:
- M/F ratio per cage: 1:1
- Length of cohabitation: 14 days
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy.
- After successful mating each pregnant female was caged (how): After confirmation on presence of sperm in the vaginal smear (Day 0 of pregnancy), the mated pairs were separated. Males were housed with their former cage mates while females were housed individually. Sterilized paper shreds were provided as a nesting material for main group females from gestation day 20 onwards.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Homogeneity and dose formulation analysis for dose concentration verification was performed during week 1 and 5 of the treatment. The results of dose concentration verifications for the prepared test item formulations and the mean results of analysis were within the range of 85 to 115% of the nominal concentration and the relative standard deviation (% RSD) was less than 15%.
Duration of treatment / exposure:
The test item or vehicle were administered to the animals through the oral (gavage) route once daily.
The males were treated for two weeks pre-mating, during mating period and up to the day before sacrifice (total of completion of 31 days of administration).
The females were treated for a two-week pre-mating period (14-days), during mating, pregnancy (gestation) and up to lactation day 13 (ranging from a total period of 50 to 62 days).
The females with positive mating signal, but no-evidence of pregnancy / littering were treated during two-week pre-mating period, during mating and further 24 days from the day of positive mating signal (group G1 - one female until experimental day 39; group G2 - one female until experimental day 52, one female until experimental day 46; G3 - two females until experimental day 49; group G4 - one female until experimental day 39).
The recovery group animals were treated until the first scheduled sacrifice of dams (total of 52 days of administration) and kept without treatment for further 18-days of observation period.
The test item/vehicle was administered by oral (gavage) route using a stainless-steel intubation cannula attached to a disposable syringe. All the doses were administered in an equivolume of 10 mL/kg body weight/day with the concentration of 11.1, 33.3 and 100 mg/mL for low, mid, and high dose groups, respectively. Vehicle was administered to the vehicle control group at an equivolume of 10 mL/kg body weight/day.
The actual dose volume for each animal was calculated based on the most recent body weight. The test item formulations were administered as soon as possible after preparation.
The prepared formulations were stirred using magnetic stirrer during administration to maintain homogeneity.
Frequency of treatment:
Once Daily
Details on study schedule:
- Age at mating of the mated animals in the study: 13 to 14 weeks
Dose / conc.:
0 mg/kg bw/day
Remarks:
Vehicle Control
Dose / conc.:
111 mg/kg bw/day
Remarks:
Low Dose
Dose / conc.:
333 mg/kg bw/day
Remarks:
Mid Dose
Dose / conc.:
1 000 mg/kg bw/day
Remarks:
High Dose
No. of animals per sex per dose:
Main Group: 24 (12 Males + 12 Females)
Recovery Group: 10 (5 Males + 5 Females)
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: There was no information or data available from repeated dose/reproduction toxicity and teratology studies for the test item.
Based on the above available information, the doses of 0, 111, 333 and 1000 mg/kg body weight/day were selected as control, low, mid and high dose groups for the dose range finding study (BIO-DTX 056) in consultation with the sponsor.
There were no test item-related related changes noted in any of the systemic, reproduction and developmental parameters up to the dose level of 1000 mg/kg body weight/day in the Dose Range Finding Study (BIO-DTX 056).
Hence, the same doses of 0, 111, 333 and 1000 mg/kg body weight/day were selected as control, low, mid and high dose groups respectively, for the present study in consultation with the sponsor.

- Rationale for animal assignment: The animals were weighed and arranged in ascending order of their body weights. These body weight stratified animals were distributed to all the main and recovery groups using Microsoft Excel Spreadsheet, such that body weight variation of animals selected for the study does not exceed ±20% (Males: -6.80 to 11.14%; Females: -12.58 to 8.29%) of the mean body weight of each sex. The grouping was done one day prior to initiation of treatment. Body weight of the animals were analyzed statistically for mean body weight to rule out statistical significant differences between groups within each sex.
Parental animals: Observations and examinations:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once daily for general clinical signs of toxicity.
- Cage side observations checked in table [No.1] were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Day 1 before treatment and weekly thereafter during treatment.

BODY WEIGHT: Yes
- Time schedule for examinations: The main group males were weighed at receipt, on the first day of dosing, weekly thereafter and at termination.
The main group females were weighed at receipt, on the first day of dosing, weekly thereafter during pre-mating and until confirmation of mating. All the pregnant animals were weighed on gestation days (GD) 0, 7, 14 and 20 and on lactation days (LD) 1, 4, 7, 13 and 14 (terminal body weight). The non-pregnant animals were weighed weekly once until termination and the terminal body weights were recorded before scheduled sacrifice.

The recovery group animals were weighed at receipt, on the first day of dosing, weekly thereafter and at termination.
Oestrous cyclicity (parental animals):
Oestrus cyclicity was monitored for two weeks after five days of acclimatization to evaluate the normal oestrus cycle (4 to 5 days). Females with normal oestrus cyclicity only were selected for the treatment. Vaginal smears were monitored daily from beginning of the treatment period until evidence of mating. When obtaining vaginal/cervical cells, care was taken to avoid disturbance of mucosa, which may induce pseudopregnancy. The status of oestrus cyclicity of females was determined on termination day.

Recovery group females were not evaluated for oestrus cyclicity
Sperm parameters (parental animals):
Parameters examined in all males: testis weight, epididymis weight.
Litter observations:
STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no

PARAMETERS EXAMINED
The following parameters were examined in [F1] offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities, anogenital distance (AGD), presence of nipples/areolae in male pups.

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities; possible cause of death was not determined for pups born or found dead.
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All animals were sacrificed after completion of 31 days of treatment. The males were necropsied in randomized manner.
- Maternal animals: All pregnant females were sacrificed on lactation day 14 and the non-pregnant females were sacrificed after 24 days from the last day of confirmation of mating.

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.

HISTOPATHOLOGY / ORGAN WEIGHTS
The tissues indicated in Table [22] were prepared for microscopic examination and weighed, respectively.
Postmortem examinations (offspring):
SACRIFICE
- Dead pups and pups which were sacrificed on PND 4/13, examined for gross abnormalities with particular attention to the external reproductive genitals and the findings were recorded.

GROSS NECROPSY
- Gross necropsy consisted of external and internal examinations including the cervical, thoracic, and abdominal viscera.
Statistics:
The raw data was subjected to computer statistical processing. The computer printout of the data (in the form of an appendix) was verified with the raw data. After verification, the data was subjected to various statistical analyses using SPSS software version 22.
All analysis and comparisons were evaluated at 95% with the level of confidence of P<0.05 respectively, indicated by the aforementioned tests and were designated by the superscripts throughout the report as stated below:
* Statistically significant (P<0.05) change than the vehicle control group.
Note: Data of non-pregnant females was excluded for mean calculations and statistical analysis for reproductive end points but considered for systemic parameters. However, the individual animal data is presented in appendices.
The statistical analysis tests followed for the interpretation of data are as mentioned below:

Parametric - One-way ANOVA
• Body weight (weekly/gestation/lactation)
• Percent Change in body weight (weekly/gestation/lactation)
• Feed consumption (weekly/gestation/lactation)
• Copulatory interval
• Gestation length
• Hematology
• Clinical chemistry
• Urinalysis
• Absolute/relative organ weights
• Functional Observation parameters
• Mean pup weight per litter
• Mean pup anogenital distance ratio per litter
• Serum T4 values (adults/pups).

Non Parametric - Kruskal-Wallis
• Implantations/litter
• No. of pups/litter
• Sex ratio/litter
• Litter size
• No. of early resorptions/litter
• No. of late resorptions/litter
• Prenatal loss/litter
• Postnatal loss/litter

Cross Tabs - Chi-square test
• Reproductive performances (frequency occurrences)
- mated/non-mated;
- fertile/infertile;
- pregnant/non-pregnant;
- with/without live pups;
- with/without dead pups


Reproductive indices:
The following reproductive performance indices were calculated as mentioned below and presented in the study report,

Male Mating Index (%) = No. of males with confirmed mating / Total No. of males cohabited x 100
Female Mating Index (%) = No. of sperm-positive females / Total No. of females cohabited x 100
Male Fertility Index (%) = No. of males impregnating a female / Total No. of males mated x 100
Female Fertility Index (%) = No. of pregnant females / No. of sperm-positive females x 100
Pre-coital Interval (Days) = Date of confirmation of mating – Date of initiation of cohabitation
Gestation Length (days) = Date of parturition – Date of positive evidence of mating (GD 0)
Gestation Index (%) = No. of females with live born / No. of females with evidence of pregnancy x 100
Parturition Index (%) = No. of females littered / No. of females with evidence of pregnancy x 100
Pregnancy Index (%) = No. of pregnant females / No. of females with confirmed mating x 100
Offspring viability indices:
Sex ratio (m/f) = No. of male offspring / Number of female offspring
Live Birth Index (%) per litter = No. of pups born alive / Total No. of pups born x 100
Pup Survival index (%) on LD 4/7/13 = Total No. of live pups on LD 4/7/13 / No. of pups born/4/7/13 x 100
Sex ratio (m/f) on LD 4/7/13 = No. of male offspring on LD 4/7/13 / No. of female offspring on LD 4/7/13
Post-Implantation Loss (%) = No. of implantations - No. of viable pups / No. of implantations x 100
Post-implantation Loss (No.) = No. of implantations - No. of live births
Postnatal loss on LD 13 (%) = No. of pups alive on postnatal day 13 /No. of live pups at birth x 100
Postnatal Loss on LD 13 (No.) = Live births - pups alive at lactation day 13
Anogenital Distance (AGD) Ratio = Anogenital distance (mm) / Cube root of PND 4 pup weight x 100
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
no effects observed
Food efficiency:
no effects observed
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
no effects observed
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Other effects:
no effects observed
Reproductive function: oestrous cycle:
no effects observed
Reproductive performance:
no effects observed
Key result
Dose descriptor:
NOAEL
Effect level:
ca. 1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food efficiency
ophthalmological examination
haematology
clinical biochemistry
urinalysis
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic
reproductive function (oestrous cycle)
reproductive function (sperm measures)
reproductive performance
other: Thyroid hormonal levels
Key result
Critical effects observed:
no
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
no effects observed
Gross pathological findings:
no effects observed
Other effects:
effects observed, non-treatment-related
Description (incidence and severity):
There were no changes in mean serum T4 hormone levels of pups at postnatal day 13 in any of the tested dose group litters when compared with vehicle control group litters. However, an incidental statistically significant decrease in mean T4 levels in pups was noted in groups G2, G3 and G4 when compared with vehicle control group.
Description (incidence and severity):
The oral administration of test item Hostaperm Red Violet ER 02 over the time of the study, did not produce any indication of developmental toxicity at the dose levels of 111, 333 and 1000 mg/kg body weight/day.
Key result
Dose descriptor:
NOAEL
Generation:
F1
Effect level:
ca. 1 000 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
viability
clinical signs
mortality
body weight and weight gain
gross pathology
other: Serum Thyroid hormonal levels (T4)
Key result
Critical effects observed:
no
Key result
Reproductive effects observed:
no

TABLE 1.     SUMMARY OF CLINICAL SIGNS OF TOXICITY, DETAILED CLINICAL EXAMINATION AND MORTALITY RECORD

Group, Sex & Dose

(mg/kg body weight/day)

Total No. of Animals

Clinical Signs of Toxicitya:

Observation
(No. of Animals revealed)

Detailed Clinical Examinationb:

Observation
(No. of Animals revealed)

Mortalityc:

No. of Mortalities
(Total No. of Animals)

G1, M & 0

12

N (12)

NAD (12)

0 (12)

G2, M & 111

12

N (12)

NAD (12)

0 (12)

G3, M & 333

12

N (12)

NAD (12)

0 (12)

G4, M & 1000

12

N (12)

NAD (12)

0 (12)

M: Male; N: Normal; NAD: No Abnormality Detected.
a: observed once daily; b: observed once weekly; c: observed twice daily.

Note:Dark purple colored faeces were noted from day 2 observation throughout the experiment period till termination in group G3 & G4 animals.

TABLE 1 (Contd…). SUMMARY OF CLINICAL SIGNS OF TOXICITY, DETAILED CLINICAL EXAMINATION AND MORTALITY RECORD

Group, Sex & Dose

(mg/kg body weight/day)

Total No. of Animals

Clinical Signs of Toxicitya:

Observation
(No. of Animals revealed)

Detailed Clinical Examinationb:

Observation
(No. of Animals revealed)

Mortalityc:

No. of Mortalities
(Total No. of Animals)

G1, F & 0

12

N (12)

NAD (12)

0 (12)

G2, F & 111

12

N (12)

NAD (12)

0 (12)

G3, F & 333

12

N (12)

NAD (12)

0 (12)

G4, F & 1000

12

N (12)

NAD (12)

0 (12)

F: Female; N: Normal; NAD: No Abnormality Detected.
a: observed once daily; b: observed once weekly; c: observed twice daily.

Note:Dark purple colored faeces were noted from day 2 observation throughout the experiment period till termination in group G3 & G4 animals.

TABLE 1 (Contd…). SUMMARY OF CLINICAL SIGNS OF TOXICITY, DETAILED CLINICAL EXAMINATION AND MORTALITY RECORD

Group, Sex & Dose

(mg/kg body weight/day)

Total No. of Animals

Clinical Signs of Toxicitya:

Observation
(No. of Animals revealed)

Detailed Clinical Examinationb:

Observation
(No. of Animals revealed)

Mortalityc:

No. of Mortalities
(Total No. of Animals)

G1R, M & 0

5

N (5)

NAD (5)

0 (5)

G4R, M & 1000

5

N (5)

NAD (5)

0 (5)

G1R, F & 0

5

N (5)

NAD (5)

0 (5)

G4R, F & 1000

5

N (5)

NAD (5)

0 (5)

M : Male; F: Female; R: Recovery; N: Normal; NAD: No abnormality detected.
a: observed once daily; b: observed once weekly; c: observed twice daily.
Note:Dark purple colored faeces were noted from day 2 throughout the experiment period till termination in group G4R animals.         
        

TABLE 2.     SUMMARY OF BODY WEIGHT (g) RECORD

Group, Sex & Dose

(mg/kg body weight/day)

 

Body Weight (g) on Day

1

7

14

21

28

G1, M & 0

Mean

400.75

421.20

438.19

446.01

458.77

±SD

22.07

32.34

36.49

37.08

38.53

n

12

12

12

12

12

G2, M & 111

Mean

401.74

417.26

433.41

442.75

452.56

±SD

20.39

24.35

30.70

31.29

30.16

n

12

12

12

12

12

G3, M & 333

Mean

402.72

419.51

438.54

444.84

456.36

±SD

19.35

23.54

30.11

27.23

24.95

n

12

12

12

12

12

G4, M & 1000

Mean

401.99

422.76

443.74

453.94

463.38

±SD

21.50

27.21

34.32

36.83

39.17

n

12

12

12

12

12

M: Male; SD: Standard Deviation; n: Number of Animals.

TABLE 2 (Contd…). SUMMARY OF BODY WEIGHT (g) RECORD

Group, Sex & Dose

(mg/kg body weight/day)

 

Body Weight (g) on Day

1

7

14

21#

G1, F & 0

Mean

250.88

257.35

267.91

280.34

±SD

14.86

17.17

18.25

26.26

n

12

12

12

3

G2, F & 111

Mean

251.05

256.43

266.53

272.56

±SD

12.78

15.62

15.30

25.98

n

12

12

12

3

G3, F & 333

Mean

251.13

256.32

261.08

260.46

±SD

13.59

14.00

15.94

17.45

n

12

12

12

4

G4, F & 1000

Mean

251.24

261.46

265.51

269.05

±SD

15.56

18.91

20.71

24.37

n

12

12

12

4

F: Female; SD: Standard Deviation; n: Number of Animals.

#: The data obtained from females in cohabitation only considered for mean calculations.

The data of day 21 body weight was not subjected to statistical analysis due to uneven number of variables.

TABLE 2 (Contd…). SUMMARY OF BODY WEIGHT (g) RECORD

Group, Sex & Dose

(mg/kg body weight/day)

 

Body Weight (g) on Day

1

7

14

21

28

35

42

49

56

63

G1R, M & 0

Mean

395.45

407.17

422.63

436.11

448.75

458.35

473.23

489.33

502.87

514.37

±SD

21.72

21.10

31.29

32.45

32.78

34.62

33.67

33.58

34.92

37.99

n

5

5

5

5

5

5

5

5

5

5

G4R, M & 1000

Mean

396.36

412.35

430.15

444.31

454.98

464.82

475.04

493.68

508.20

519.69

±SD

20.86

20.17

12.53

16.96

17.27

14.72

15.50

9.28

11.65

11.69

n

5

5

5

5

5

5

5

5

5

5

G1R, F & 0

Mean

245.24

250.97

262.44

268.72

277.56

284.92

295.96

306.72

315.86

324.75

±SD

10.97

10.11

13.68

15.80

15.60

15.72

19.40

19.59

17.41

15.21

n

5

5

5

5

5

5

5

5

5

5

G4R, F & 1000

Mean

245.66

248.99

264.41

270.43

278.43

286.33

298.35

308.32

318.51

327.87

±SD

12.82

12.43

13.84

12.16

11.19

12.10

12.79

13.72

10.97

12.44

n

5

5

5

5

5

5

5

5

5

5

M: Male; F: Female; R: Recovery; SD: Standard Deviation; n: Number of Animals.

TABLE 3.     SUMMARY RECORD OF GESTATION BODY WEIGHT (g)

Group, Sex

& Dose

(mg/kg body

weight/day)

Body Weight (g) on Gestation Day (GD)

0

7

14

20

G1, F & 0

Mean

272.42

287.38

319.92

375.62

±SD

18.33

21.41

28.89

35.87

n

11

11

11

11

G2, F & 111

Mean

269.81

286.87

328.45

391.71

±SD

11.98

12.37

15.11

22.46

n

10

10

10

10

G3, F & 333

Mean

264.91

282.42

317.51

369.85

±SD

15.69

14.58

20.68

32.90

n

10

10

10

10

G4, F & 1000

Mean

273.49

291.35

327.07

382.89

±SD

17.37

17.37

23.26

39.30

n

11

11

11

11

F: Female; SD: Standard Deviation; n: Number of Dams.

Note: Excluded the data of non-pregnant females for mean calculations and statistical analysis, but presented in individual animal data.

TABLE 4.     SUMMARY RECORD OF LACTATION BODY WEIGHT (g)

Group, Sex

& Dose

(mg/kg body

weight/day)

Body Weight (g) on Lactation Day (LD)

1

4

7

13

G1, F & 0

Mean

294.60

300.48

308.18

323.25

±SD

22.07

22.28

22.06

20.39

n

11

11

11

11

G2, F & 111

Mean

306.24

309.48

314.78

330.57

±SD

22.46

21.39

20.72

19.26

n

10

10

10

10

G3, F & 333

Mean

293.35

297.31

304.48

319.65

±SD

19.47

20.23

20.46

22.22

n

10

10

10

10

G4, F & 1000

Mean

303.31

306.82

313.01

325.64

±SD

22.77

24.10

23.73

23.27

n

11

11

11

11

F: Female; SD: Standard Deviation; n: Number of Dams.

TABLE 5.     SUMMARY OF PERCENT CHANGE IN BODY WEIGHT GAIN (%) WITH RESPECT TO DAY 1 RECORD

Group, Sex & Dose

(mg/kg body weight/day)

 

Percent Change in Body Weight (%) during Day

1 to 7

1 to 14

1 to 21

1 to 28

G1, M & 0

Mean

5.01

9.25

11.19

14.36

±SD

3.10

4.96

4.86

4.98

n

12

12

12

12

G2, M & 111

Mean

3.84

7.80

10.13

12.59

±SD

2.04

3.09

3.48

3.20

n

12

12

12

12

G3, M & 333

Mean

4.15

8.89

10.50

13.42

±SD

2.15

5.14

5.14

5.61

n

12

12

12

12

G4, M & 1000

Mean

5.12

10.29

12.82

15.16

±SD

1.91

3.83

4.55

5.27

n

12

12

12

12

M: Male; SD: Standard Deviation; n: Number of Animals.

TABLE 5 (Contd…). SUMMARY OF PERCENTCHANGE IN BODY WEIGHT GAIN (%) WITH RESPECT TO DAY 1 RECORD

Group, Sex & Dose

(mg/kg body weight/day)

 

Percent Change in Body Weight (%) during Day

1 to 7

1 to 14

1 to 21#

G1, F & 0

Mean

2.56

6.78

10.07

±SD

2.21

3.23

3.01

n

12

12

3

G2, F & 111

Mean

2.12

6.21

9.94

±SD

2.46

4.16

1.49

n

12

12

3

G3, F & 333

Mean

2.08

3.94

5.98

±SD

1.81

2.08

1.64

n

12

12

4

G4, F & 1000

Mean

4.05

5.61

9.37

±SD

3.24

3.20

3.75

n

12

12

4

F: Female; SD: Standard Deviation; n: Number of Animals.

#: The data obtained from females in cohabitation only considered for mean calculations. The data of day 1 to 21 body weight was not subjected to statistical analysis due to uneven number of variables.

TABLE 5 (Contd…). SUMMARY OF PERCENTCHANGE IN BODY WEIGHT GAIN (%) WITH RESPECT TO DAY 1 RECORD

Group, Sex & Dose

(mg/kg body weight/day)

 

Percent Change in Body Weight (%) during Day

1 to 7

1 to 14

1 to 21

1 to 28

1 to 35

1 to 42

1 to 49

1 to 56

1 to 63

G1R, M & 0

Mean

2.98

6.83

10.25

13.47

15.92

19.69

23.73

27.13

30.00

±SD

0.92

4.09

4.67

5.29

6.27

5.81

4.88

4.25

4.30

n

5

5

5

5

5

5

5

5

5

G4R, M & 1000

Mean

4.06

8.66

12.20

14.91

17.44

20.09

24.77

28.45

31.32

±SD

1.73

3.60

3.33

3.84

4.90

6.78

5.51

6.10

5.14

n

5

5

5

5

5

5

5

5

5

G1R, F & 0

Mean

2.35

7.00

9.55

13.20

16.19

20.62

25.02

28.77

32.44

±SD

0.72

2.36

3.15

4.61

4.45

4.14

4.54

3.55

3.44

n

5

5

5

5

5

5

5

5

5

G4R, F & 1000

Mean

1.37

7.66

10.20

13.45

16.65

21.52

25.60

29.78

33.57

±SD

1.07

2.71

4.80

4.07

3.73

3.02

4.10

3.75

3.31

n

5

5

5

5

5

5

5

5

5

M: Male; F: Female; R: Recovery; SD: Standard Deviation; n: Number of Animals.

TABLE 6.     SUMMARY RECORD OF PERCENT CHANGE IN BODY WEIGHT GAIN (%) DURING GESTATION PERIOD

Group, Sex

& Dose

(mg/kg body

weight/day)

Percent Change in Body Weight Gain (%) during Gestation Day (GD)

0 to 7

7 to 14

14 to 20

G1, F & 0

Mean

5.46

11.22

17.44

±SD

1.77

2.82

4.18

n

11

11

11

G2, F & 111

Mean

6.35

14.52

19.23

±SD

2.12

3.20

3.16

n

10

10

10

G3, F & 333

Mean

6.67

12.40

16.35

±SD

1.92

3.55

3.43

n

10

10

10

G4, F & 1000

Mean

6.56

12.23

16.88

±SD

1.60

3.35

4.81

n

11

11

11

F: Female; SD: Standard Deviation; n: Number of Dams.

Note: Excluded the data of non-pregnant females for mean calculations and statistical analysis, but presented in individual animal data.

TABLE 7.     SUMMARY RECORD OF PERCENT CHANGE IN BODY WEIGHT GAIN (%) DURING LACTATION PERIOD

Group, Sex

& Dose

(mg/kg body

weight/day)

Percent Change in Body Weight Gain (%) during Lactation Day (LD)

1 to 4

4 to 7

7 to 13

G1, F & 0

Mean

2.01

2.58

4.96

±SD

0.96

0.73

1.51

n

11

11

11

G2, F & 111

Mean

1.08

1.74

5.07

±SD

0.86

0.95

1.60

n

10

10

10

G3, F & 333

Mean

1.35

2.42

4.98

±SD

0.95

1.29

1.32

n

10

10

10

G4, F & 1000

Mean

1.14

2.04

4.06

±SD

1.00

0.93

0.83

n

11

11

11

F: Female; SD: Standard Deviation; n: Number of Dams.

TABLE 8.     SUMMARY OF FEED CONSUMPTION (g/animal/day) RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Feed Consumption (g/animal/day) during Pre-mating Period

Week 1

Week 2

G1, M & 0

Mean

21.31

24.01

±SD

2.38

1.81

n

12 (6)

12 (6)

G2, M & 111

Mean

21.39

23.12

±SD

1.76

0.75

n

12 (6)

12 (6)

G3, M & 333

Mean

22.25

23.93

±SD

2.39

1.85

n

12 (6)

12 (6)

G4, M & 1000

Mean

22.28

24.17

±SD

1.23

0.67

n

12 (6)

12 (6)

M: Male; SD: Standard Deviation; n: Number of Animals (Number of cages).

TABLE 8 (Contd…). SUMMARY OF FEED CONSUMPTION (g/animal/day) RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Feed Consumption (g/animal/day) during Pre-mating Period

Week 1

Week 2

G1, F & 0

Mean

18.14

19.17

±SD

1.56

1.22

n

12 (6)

12 (6)

G2, F & 111

Mean

17.79

20.11

±SD

1.85

1.70

n

12 (6)

12 (6)

G3, F & 333

Mean

18.26

19.22

±SD

1.99

1.31

n

12 (6)

12 (6)

G4, F & 1000

Mean

18.43

20.07

±SD

2.28

1.22

n

12 (6)

12 (6)

F: Female; SD: Standard Deviation; n: Number of Animals (Number of cages).

TABLE 8 (Contd…). SUMMARY OF FEED CONSUMPTION (g/animal/day) RECORD

Group, Sex

& Dose

(mg/kg body weight/day)

 

Feed Consumption (g/animal/day)

Week 1

Week 2

Week 3

Week 4

Week 5

Week 6

Week 7

Week 8

Week 9

G1R, M & 0

Mean

23.52

24.34

25.11

26.35

27.86

29.05

30.75

31.83

32.47

±SD

3.71

1.67

1.55

1.27

0.97

1.11

0.47

3.66

3.01

n

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

G4R, M & 1000

Mean

22.84

23.75

24.26

27.17

28.86

30.05

31.07

32.19

33.03

±SD

0.77

1.04

1.34

2.09

2.61

2.18

2.12

1.59

1.67

n

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

G1R, F & 0

Mean

17.44

18.36

19.45

10.21

23.17

24.20

25.35

26.19

27.04

±SD

1.79

0.40

0.40

9.72

0.99

0.82

0.62

0.41

0.99

n

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

G4R, F & 1000

Mean

17.21

19.05

20.42

11.29

24.18

25.37

26.19

27.14

27.89

±SD

0.69

1.77

0.91

10.38

0.03

0.80

0.84

0.69

0.68

n

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

5 (3)

M: Male; F: Female; R: Recovery; SD: Standard Deviation; n: Number of Animals(Number of cages).

TABLE 9.     SUMMARY RECORD OF FEED CONSUMPTION (g/animal/day) DURING GESTATION PERIOD

Group, Sex

& Dose

(mg/kg body

weight/day)

 

Feed Consumption (g/animal/day) during Gestation Day (GD)

0 to 7

7 to 14

14 to 20

G1, F & 0

Mean

19.02

21.07

25.81

SD

0.70

1.95

1.63

n

11

11

11

G2, F & 111

Mean

19.23

22.20

26.84

SD

0.88

1.81

1.78

n

10

10

10

G3, F & 333

Mean

19.00

21.49

26.11

SD

0.74

1.39

1.26

n

10

10

10

G4, F & 1000

Mean

19.43

22.01

26.67

SD

0.58

1.70

1.97

n

11

11

11

F: Female; SD: Standard Deviation; n: Number of Dams.

Note: Excluded the data of non-pregnant females for mean calculations and statistical analysis but presented in individual animal data.

TABLE 10.     SUMMARY RECORD OF FEED CONSUMTION (g/animal/day) DURING LACTATION PERIOD

Group, Sex

& Dose

(mg/kg body

weight/day)

 

Feed Consumption (g/animal/day) during Lactation Day (LD)

1 to 4

4 to 7

7 to 13

G1, F & 0

Mean

25.21

35.80

40.88

SD

1.20

7.26

1.43

n

11

11

11

G2, F & 111

Mean

24.58

32.72

41.09

SD

1.34

0.99

0.98

n

10

10

10

G3, F & 333

Mean

25.11

32.75

40.54

SD

1.42

1.16

0.94

n

10

10

10

G4, F & 1000

Mean

23.94

33.14

39.84

SD

5.99

1.28

1.03

n

11

11

11

F: Female; SD: Standard Deviation; n: Number of Dams.

TABLE 11.     SUMMARY OF OPHTHALMOLOGICAL EXAMINATION RECORD

Day -1 (Grouping and randomization)

Group, Sex

& Dose
(mg/kg body

weight/day) →

G1, M & 0

G2, M & 111

G3, M & 333

G4, M & 1000

Number of Animals

12

12

12

12

Observations↓

Eye

Region↓

LE

RE

LE

RE

LE

RE

LE

RE

Eye Lids

N

N

N

N

N

N

N

N

Cornea

N

N

N

N

N

N

N

N

Iris

N

N

N

N

N

N

N

N

Aqueous Humour

N

N

N

N

N

N

N

N

Lens

N

N

N

N

N

N

N

N

Vitreous Humour

N

N

N

N

N

N

N

N

Retina/Optic disc

N

N

N

N

N

N

N

N

M: Male; N: Normal; LE: Left Eye; RE: Right Eye.

TABLE 11 (Contd…). SUMMARY OF OPHTHALMOLOGICAL EXAMINATION RECORD

Day -1 (Grouping and randomization)

Group, Sex

& Dose
(mg/kg body
     

weight/day) →

G1, F & 0

G2, F & 111

G3, F & 333

G4, F & 1000

Number of Animals

12

12

12

12

Observations↓

Eye

Region↓

LE

RE

LE

RE

LE

RE

LE

RE

Eye Lids

N

N

N

N

N

N

N

N

Cornea

N

N

N

N

N

N

N

N

Iris

N

N

N

N

N

N

N

N

Aqueous Humour

N

N

N

N

N

N

N

N

Lens

N

N

N

N

N

N

N

N

Vitreous Humour

N

N

N

N

N

N

N

N

Retina/Optic disc

N

N

N

N

N

N

N

N

F: Female; N: Normal; LE: Left Eye; RE: Right Eye.

TABLE 11 (Contd…). SUMMARY OF OPHTHALMOLOGICAL EXAMINATION RECORD

Day -1 (Grouping and randomization)

Group, Sex

& Dose
(mg/kg body
     

weight/day) →

G1R, M & 0

G4R, M & 1000

Number of Animals

5

5

Observations↓

Eye

Region↓

LE

RE

LE

RE

Eye Lids

N

N

N

N

Cornea

N

N

N

N

Iris

N

N

N

N

Aqueous Humour

N

N

N

N

Lens

N

N

N

N

Vitreous Humour

N

N

N

N

Retina/Optic disc

N

N

N

N

M: Male; R: Recovery; N: Normal; LE: Left Eye; RE: Right Eye.

TABLE 11 (Contd…). SUMMARY OF OPHTHALMOLOGICAL EXAMINATION RECORD

Day -1 (Grouping and randomization)

Group, Sex

& Dose
(mg/kg body
     

weight/day) →

G1R, F & 0

G4R, F & 1000

Number of Animals

5

5

Observations↓

Eye

Region↓

LE

RE

LE

RE

Eye Lids

N

N

N

N

Cornea

N

N

N

N

Iris

N

N

N

N

Aqueous Humour

N

N

N

N

Lens

N

N

N

N

Vitreous Humour

N

N

N

N

Retina/Optic disc

N

N

N

N

F: Female; R: Recovery; N: Normal; LE: Left Eye; RE: Right Eye.

TABLE 11 (Contd…). SUMMARY OF OPHTHALMOLOGICAL EXAMINATION RECORD

Day 30

Group, Sex

& Dose
(mg/kg body

weight/day) →

G1, M & 0

G2, M & 111

G3, M & 333

G4, M & 1000

Number of Animals

12

12

12

12

Observations↓

Eye

Region↓

LE

RE

LE

RE

LE

RE

LE

RE

Eye Lids

N

N

N

N

N

N

N

N

Cornea

N

N

N

N

N

N

N

N

Iris

N

N

N

N

N

N

N

N

Aqueous Humour

N

N

N

N

N

N

N

N

Lens

N

N

N

N

N

N

N

N

Vitreous Humour

N

N

N

N

N

N

N

N

Retina/Optic disc

N

N

N

N

N

N

N

N

Other findings:

N

N

N

N

N

N

N

N

M: Male; N: Normal; LE: Left Eye; RE: Right Eye.

TABLE 11 (Contd…). SUMMARY OF OPHTHALMOLOGICAL EXAMINATION RECORD

Pregnant/Littered females: Lactation day 13;

Non-pregnant females: (Experimental day 39 to 52)

Group, Sex

& Dose
(mg/kg body

weight/day) →

G1, F & 0

G2, F & 111

G3, F & 333

G4, F & 1000

Number of Animals

12

12

12

12

Observations↓

Eye

Region↓

LE

RE

LE

RE

LE

RE

LE

RE

Eye Lids

N

N

N

N

N

N

N

N

Cornea

N

N

N

N

N

N

N

N

Iris

N

N

N

N

N

N

N

N

Aqueous Humour

N

N

N

N

N

N

N

N

Lens

N

N

N

N

N

N

N

N

Vitreous Humour

N

N

N

N

N

N

N

N

Retina/Optic disc

N

N

N

N

N

N

N

N

Other findings:

N

N

N

N

N

N

N

N

F: Female; N: Normal; LE: Left Eye; RE: Right Eye.

TABLE 11 (Contd…). SUMMARY OF OPHTHALMOLOGICAL EXAMINATION RECORD

Day 65

Group, Sex

& Dose
(mg/kg body

weight/day) →

G1R, M & 0

G4R, M & 1000

Number of Animals

5

5

Observations↓

Eye

Region↓

LE

RE

LE

RE

Eye Lids

N

N

N

N

Cornea

N

N

N

N

Iris

N

N

N

N

Aqueous Humour

N

N

N

N

Lens

N

N

N

N

Vitreous Humour

N

N

N

N

Retina/Optic disc

N

N

N

N

Other findings:

N

N

N

N

M: Male; R: Recovery; N: Normal; LE: Left Eye; RE: Right Eye.

TABLE 11 (Contd…). SUMMARY OF OPHTHALMOLOGICAL EXAMINATION RECORD

Day 65

Group, Sex

& Dose
(mg/kg body

weight/day) →

G1R, F & 0

G4R, F & 1000

Number of Animals

5

5

Observations↓

Eye

Region↓

LE

RE

LE

RE

Eye Lids

N

N

N

N

Cornea

N

N

N

N

Iris

N

N

N

N

Aqueous Humour

N

N

N

N

Lens

N

N

N

N

Vitreous Humour

N

N

N

N

Retina/Optic disc

N

N

N

N

Other findings:

N

N

N

N

F: Female; R: Recovery; N: Normal; LE: Left Eye; RE: Right Eye.

TABLE 12.     SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB) RECORD

Day 30

Parameters↓

Group & Sex

G1 & M

G2 & M

G3 & M

G4 & M

Dose (mg/kg body weight/day)

0

111

333

1000

Number of Animals (randomly selected)

5

5

5

5

Home Cage Observations

Home cage posture

1

1

1

1

Respiratory pattern

1

1

1

1

Clonic involuntary movements

1

1

1

1

Tonic involuntary movements

1

1

1

1

Vocalization

1

1

1

1

Palpebral closure

1

1

1

1

Handling Observations

Ease of removal from the cage

2

2

2

2

Ease of handling animal in hand

2

2

2

2

Red or crusty deposits

Eyes

1

1

1

1

Nose

1

1

1

1

Mouth

1

1

1

1

Lacrimation

1

1

1

1

Salivation

1

1

1

1

Fur appearance

1

1

1

1

Piloerection

1

1

1

1

Eye prominence

1

1

1

1

Muscle tone

1

1

1

1

Home Cage Observations:Home cage posture - 1=Normal; Respiratory pattern - 1=Normal; Clonic involuntary movements - 1=None; Tonic involuntary movements - 1=None; Vocalization - 1=None; Palpebral closure - 1=Normal;Handling Observations:Ease of removal from the cage - 2=Normal; Ease of handling animal in hand - 2=Normal; Red or crusty deposits - 1=Absent; Lacrimation - 1=None; Salivation- 1=None; Fur appearance - 1=Normal; Piloerection - 1=None; Eye Prominence - 1=Normal; Muscle tone - 1=Normal

TABLE 12 (Contd…). SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB) RECORD

Day 30

Parameters↓

Group & Sex

G1 & M

G2 & M

G3 & M

G4 & M

Dose (mg/kg body weight/day)

0

111

333

1000

Number of Animals (randomly selected)

5

5

5

5

Open Field Observations

Mobility

1

1

1

1

Gait

1

1

1

1

Arousal

3

3

3

3

Number of rearing

Mean

4.4

4.8

4.8

4.8

±SD

1.1

0.8

0.8

0.8

Number of urine pools

Mean

3.4

3.8

3.8

3.8

±SD

0.5

0.8

0.8

0.8

Number of defecations

Mean

3.4

3.2

3.4

3.2

±SD

0.5

0.8

0.5

0.4

Clonic involuntary movement

1

1

1

1

Tonic involuntary movement

1

1

1

1

Stereotype behaviour

1

1

1

1

Number of grooming

Mean

3.4

3.2

3.4

3.2

±SD

0.5

0.8

0.5

0.8

Sensory Observations

Approach response

2

2

2

2

Auditory response

2

2

2

2

Touch response

2

2

2

2

Pupil reflex

2

2

2

2

Tail pinch response

2

2

2

2

Righting reflex

1

1

1

1

Physiological Observation

Body temperature (°F)

Mean

98.4

98.4

98.1

98.4

±SD

0.5

0.4

0.3

0.5

Open Field Observations: Mobility- 1=Normal;Gait- 1=Normal;Arousal- 3=Normal;Clonic involuntary movement- 1=None;Tonic involuntary movement -1=None;Stereotype behaviour -1=Absent;Sensory Observations:Approach response- 2=Normal;Auditory response- 2=Normal;Touch response -2=Normal;Pupil reflex- 2=Normal;Tail pinch response- 2=Normal;Righting reflex- 1=Present

TABLE 12 (Contd…). SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB)

Lactation Day 13

Parameters↓

Group & Sex

G1 & F

G2 & F

G3 & F

G4 & F

Dose (mg/kg body weight/day)

0

111

333

1000

Number of Animals (randomly selected)

5

5

5

5

Home Cage Observations

Home cage posture

1

1

1

1

Respiratory pattern

1

1

1

1

Clonic involuntary movements

1

1

1

1

Tonic involuntary movements

1

1

1

1

Vocalization

1

1

1

1

Palpebral closure

1

1

1

1

Handling Observations

Ease of removal from the cage

2

2

2

2

Ease of handling animal in hand

2

2

2

2

Red or crusty deposits

Eyes

1

1

1

1

Nose

1

1

1

1

Mouth

1

1

1

1

Lacrimation

1

1

1

1

Salivation

1

1

1

1

Fur appearance

1

1

1

1

Piloerection

1

1

1

1

Eye prominence

1

1

1

1

Muscle tone

1

1

1

1

Home Cage Observations:Home cage posture - 1=Normal; Respiratory pattern - 1=Normal; Clonic involuntary movements - 1=None; Tonic involuntary movements - 1=None; Vocalization - 1=None; Palpebral closure - 1=Normal;Handling Observations:Ease of removal from the cage - 2=Normal; Ease of handling animal in hand - 2=Normal; Red or crusty deposits - 1=Absent; Lacrimation - 1=None; Salivation- 1=None; Fur appearance - 1=Normal; Piloerection - 1=None; Eye Prominence - 1=Normal; Muscle tone - 1=Normal

TABLE 12 (Contd…). SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB)

Lactation Day 13

Parameters↓

Group & Sex

G1 & F

G2 & F

G3 & F

G4 & F

Dose (mg/kg body weight/day)

0

111

333

1000

Number of Animals (randomly selected)

5

5

5

5

Open Field Observations

Mobility

1

1

1

1

Gait

1

1

1

1

Arousal

3

3

3

3

Number of rearing

Mean

4.8

4.8

4.8

4.8

±SD

0.8

0.8

0.8

0.8

Numbers of urine pools

Mean

3.6

3.8

3.6

3.8

±SD

0.5

0.8

0.5

0.8

Number of defecations

Mean

3.4

3.0

2.8

3.2

±SD

0.9

0.7

0.8

0.4

Clonic involuntary movement

 1

 1

 1

 1

Tonic involuntary movement

 1

 1

 1

 1

Stereotype behaviour

 1

 1

 1

 1

Number of grooming

Mean

3.2

3.2

2.6

3.0

±SD

0.4

0.8

0.5

0.7

Sensory Observations

Approach response

2

2

2

2

Auditory response

2

2

2

2

Touch response  

2

2

2

2

Pupil reflex

2

2

2

2

Tail pinch response

2

2

2

2

Righting reflex

1

1

1

1

Physiological Observation

Body temperature (°F)

Mean

98.4

98.4

98.8

98.7

±SD

0.6

0.5

0.5

0.8

Open Field Observations: Mobility- 1=Normal;Gait- 1=Normal;Arousal- 3=Normal;Clonic involuntary movement- 1=None;Tonic involuntary movement -1=None;Stereotype behaviour -1=Absent;Sensory Observations:Approach response- 2=Normal;Auditory response- 2=Normal;Touch response -2=Normal;Pupil reflex- 2=Normal;Tail pinch response- 2=Normal;Righting reflex- 1=Present

TABLE 12 (Contd…). SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB)

Day 65

Parameters↓

Group & Sex

G1R & M

G4R & M

Dose (mg/kg body weight/day)

0

1000

Number of Animals

5

5

Home Cage Observations

Home cage posture

1

1

Respiratory pattern

1

1

Clonic involuntary movements

1

1

Tonic involuntary movements

1

1

Vocalization

1

1

Palpebral closure

1

1

Handling Observations

Ease of removal from the cage

2

2

Ease of handling animal in hand

2

2

Red or crusty deposits

Eyes

1

1

Nose

1

1

Mouth

1

1

Lacrimation

1

1

Salivation

1

1

Fur appearance

1

1

Piloerection

1

1

Eye prominence

1

1

Muscle tone

1

1

Home Cage Observations:Home cage posture - 1=Normal; Respiratory pattern - 1=Normal; Clonic involuntary movements - 1=None; Tonic involuntary movements - 1=None; Vocalization - 1=None; Palpebral closure - 1=Normal;Handling Observations:Ease of removal from the cage - 2=Normal; Ease of handling animal in hand - 2=Normal; Red or crusty deposits - 1=Absent; Lacrimation - 1=None; Salivation- 1=None; Fur appearance - 1=Normal; Piloerection - 1=None; Eye Prominence - 1=Normal; Muscle tone - 1=Normal

TABLE 12 (Contd…). SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB)

Day 65

Parameters↓

Group & Sex

G1R & M

G4R & M

Dose (mg/kg body weight/day)

0

1000

Number of Animals

5

5

Open Field Observations

Mobility

1

1

Gait

1

1

Arousal

3

3

Number of rearing

Mean

5.0

4.4

±SD

1.0

1.1

Numbers of urine pools

Mean

3.6

3.4

±SD

0.5

0.5

Number of defecations

Mean

3.6

3.4

±SD

0.5

0.5

Clonic involuntary movement

 1

 1

Tonic involuntary movement

 1

 1

Stereotype behaviour

 1

 1

Number of grooming

Mean

3.6

3.2

±SD

0.5

0.4

Sensory Observations

Approach response

2

2

Auditory response

2

2

Touch response  

2

2

Pupil reflex

2

2

Tail pinch response

2

2

Righting reflex

1

1

Physiological Observation

Body temperature (°F)

Mean

98.5

98.6

±SD

0.5

0.5

Open Field Observations: Mobility- 1=Normal;Gait- 1=Normal;Arousal- 3=Normal;Clonic involuntary movement- 1=None;Tonic involuntary movement -1=None;Stereotype behaviour -1=Absent;Sensory Observations:Approach response- 2=Normal;Auditory response- 2=Normal;Touch response -2=Normal;Pupil reflex- 2=Normal;Tail pinch response- 2=Normal;Righting reflex- 1=Present

TABLE 12 (Contd…). SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB)

Day 65

Parameters↓

Group & Sex

G1R & F

G4R & F

Dose (mg/kg body weight/day)

0

1000

Number of Animals

5

5

Home Cage Observations

Home cage posture

1

1

Respiratory pattern

1

1

Clonic involuntary movements

1

1

Tonic involuntary movements

1

1

Vocalization

1

1

Palpebral closure

1

1

Handling Observations

Ease of removal from the cage

2

2

Ease of handling animal in hand

2

2

Red or crusty deposits

Eyes

1

1

Nose

1

1

Mouth

1

1

Lacrimation

1

1

Salivation

1

1

Fur appearance

1

1

Piloerection

1

1

Eye prominence

1

1

Muscle tone

1

1

Home Cage Observations:Home cage posture - 1=Normal; Respiratory pattern - 1=Normal; Clonic involuntary movements - 1=None; Tonic involuntary movements - 1=None; Vocalization - 1=None; Palpebral closure - 1=Normal;Handling Observations:Ease of removal from the cage - 2=Normal; Ease of handling animal in hand - 2=Normal; Red or crusty deposits - 1=Absent; Lacrimation - 1=None; Salivation- 1=None; Fur appearance - 1=Normal; Piloerection - 1=None; Eye Prominence - 1=Normal; Muscle tone - 1=Normal

TABLE 12 (Contd…). SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB)

Day 65

Parameters↓

Group & Sex

G1R & F

G4R & F

Dose (mg/kg body weight/day)

0

1000

Number of Animals

5

5

Open Field Observations

Mobility

1

1

Gait

1

1

Arousal

3

3

Number of rearing

Mean

5.0

4.8

±SD

1.0

0.8

Numbers of urine pools

Mean

3.6

3.8

±SD

0.5

0.8

Number of defecations

Mean

3.4

3.0

±SD

0.9

0.7

Clonic involuntary movement

 1

 1

Tonic involuntary movement

 1

 1

Stereotype behaviour

 1

 1

Number of grooming

Mean

3.2

3.2

±SD

0.4

0.8

Sensory Observations

Approach response

2

2

Auditory response

2

2

Touch response  

2

2

Pupil reflex

2

2

Tail pinch response

2

2

Righting reflex

1

1

Physiological Observation

Body temperature (°F)

Mean

98.4

98.3

±SD

0.5

0.7

Open Field Observations: Mobility- 1=Normal;Gait- 1=Normal;Arousal- 3=Normal;Clonic involuntary movement- 1=None;Tonic involuntary movement -1=None;Stereotype behaviour -1=Absent;Sensory Observations:Approach response- 2=Normal;Auditory response- 2=Normal;Touch response -2=Normal;Pupil reflex- 2=Normal;Tail pinch response- 2=Normal;Righting reflex- 1=Present

TABLE 13.     SUMMARY OF LOCOMOTOR ACTIVITY / MOVEMENT COUNTS RECORD

Day 30

Group, Sex & Dose (mg/kg body weight/day)

Movement Counts (no.)

G1, M & 0

Mean

1911.4

±SD

108.9

n

5

G2, M & 111

Mean

1961.8

±SD

138.9

n

5

G3, M & 333

Mean

1954.2

±SD

126.1

n

5

G4, M & 1000

Mean

1931.4

±SD

104.0

n

5

M: Male; SD: Standard Deviation; n: Number of Animals (randomly selected).

TABLE 13 (Contd…). SUMMARY OF LOCOMOTOR ACTIVITY / MOVEMENT COUNTS RECORD

Lactation Day 13

Group, Sex & Dose (mg/kg body weight/day)

Movement Counts (no.)

G1, F & 0

Mean

1923.4

±SD

102.7

n

5

G2, F & 111

Mean

1941.4

±SD

125.1

n

5

G3, F & 333

Mean

1989.0

±SD

103.1

n

5

G4, F & 1000

Mean

1973.2

±SD

84.7

n

5

F: Female; SD: Standard Deviation; n: Number of Animals (randomly selected).

TABLE 13 (Contd…). SUMMARY OF LOCOMOTOR ACTIVITY / MOVEMENT COUNTS RECORD

Day 65

Group, Sex & Dose

(mg/kg body weight/day)

Movement Counts (no.)

G1R, M & 0

Mean

2014.2

±SD

210.7

n

5

G4R, M & 1000

Mean

1980.6

±SD

197.8

n

5

G1R, F & 0

Mean

1943.4

±SD

87.8

n

5

G4R, F & 1000

Mean

1956.2

±SD

91.6

n

5

M: Male; R: Recovery; SD: Standard Deviation; n: Number of Animals.

TABLE 14.     SUMMARY OF AVERAGE GRIP STRENGTH (kgf) MEASUREMENT RECORD

Day 30

Group, Sex & Dose

(mg/kg body weight/day)

Grip Strength (kgf)

Forelimb

Hindlimb

G1, M & 0

Mean

1.529

0.510

±SD

0.023

0.025

n

5

5

G2, M & 111

Mean

1.501

0.539

±SD

0.020

0.021

n

5

5

G3, M & 333

Mean

1.521

0.552*

±SD

0.048

0.008

n

5

5

G4, M & 1000

Mean

1.539

0.544*

±SD

0.040

0.015

n

5

5

M: Male; SD: Standard Deviation; n: Number of Animals (randomly selected); *: Statistically significant (P<0.05) change than the vehicle control group.

TABLE 14 (Contd…). SUMMARY OF AVERAGE GRIP STRENGTH (kgf) MEASUREMENT RECORD

Lactation Day 13

Group, Sex & Dose

(mg/kg body weight/day)

Grip Strength (kgf)

Forelimb

Hindlimb

G1, F & 0

Mean

1.496

0.508

±SD

0.009

0.008

n

5

5

G2, F & 111

Mean

1.493

0.488

±SD

0.010

0.015

n

5

5

G3, F & 333

Mean

1.496

0.493

±SD

0.009

0.021

n

5

5

G4, F & 1000

Mean

1.498

0.499

±SD

0.011

0.020

n

5

5

F: Female; SD: Standard Deviation; n: Number of Animals (randomly selected).

TABLE 14 (Contd…). SUMMARY OF AVERAGE GRIP STRENGTH (kgf) MEASUREMENT RECORD

Day 65

Group, Sex & Dose

(mg/kg body weight/day)

Grip Strength (kgf)

Forelimb

Hindlimb

G1R, M & 0

Mean

1.515

0.534

±SD

0.020

0.035

n

5

5

G4R, M & 1000

Mean

1.528

0.533

±SD

0.029

0.009

n

5

5

G1R, F & 0

Mean

1.516

0.499

±SD

0.031

0.021

n

5

5

G4R, F & 1000

Mean

1.503

0.481

±SD

0.012

0.012

n

5

5

M: Male; F: Female; R: Recovery; SD: Standard Deviation; n: Number of Animals.

TABLE 15.     SUMMARY OF AVERAGE HINDLIMB FOOT SPLAY (cm) RECORD

Day 30

Group, Sex & Dose

(mg/kg body weight/day)

Hindlimb Foot Splay (cm)

G1, M & 0

Mean

9.0

±SD

1.0

n

5

G2, M & 111

Mean

8.8

±SD

1.3

n

5

G3, M & 333

Mean

9.3

±SD

0.6

n

5

G4, M & 1000

Mean

8.6

±SD

0.6

n

5

M: Male; SD: Standard Deviation; n: Number of Animals (randomly selected).

TABLE 15 (Contd…). SUMMARY OF AVERAGE HINDLIMB FOOT SPLAY (cm) RECORD

Lactation Day 13

Group, Sex & Dose

(mg/kg body weight/day)

Hindlimb Foot Splay (cm)

G1, F & 0

Mean

8.0

±SD

0.5

n

5

G2, F & 111

Mean

7.9

±SD

0.5

n

5

G3, F & 333

Mean

8.1

±SD

1.0

n

5

G4, F & 1000

Mean

8.5

±SD

0.9

n

5

F: Female; SD: Standard Deviation; n: Number of Animals (randomly selected).

TABLE 15 (Contd…). SUMMARY OF AVERAGE HINDLIMB FOOT SPLAY (cm) RECORD

Day 65

Group, Sex & Dose

(mg/kg body weight/day)

Hindlimb Foot Splay (cm)

G1R, M & 0

Mean

8.6

±SD

0.3

n

5

G4R, M & 1000

Mean

9.0

±SD

0.4

n

5

G1R, F & 0

Mean

8.4

±SD

0.4

n

5

G4R, F & 1000

Mean

7.6*

±SD

0.2

n

5

M: Male; F: Female; R: Recovery; SD: Standard Deviation; n: Number of Animals; *: Statistically significant (P<0.05) change than the vehicle control group.

TABLE 16.     SUMMARY OF HAEMATOLOGY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Total

Leucocyte

Count

Total

Erythrocyte

Count

Haemoglobin

Haematocrit

Mean

Corpuscular

Volume

Mean

Corpuscular

Haemoglobin

Mean

Corpuscular

Haemoglobin

Concentration

Platelet

Count

Mean

Platelet

Volume

(WBC)

(RBC)

(HGB)

(HCT)

(MCV)

(MCH)

(MCHC)

(PLT)

(MPV)

(103cells/µL)

(106cells/µL)

(g/dL)

(%)

(fL)

(pg)

(g/dL)

(103cells/µL)

(fL)

G1, M & 0

Mean

8.26

9.64

17.56

52.80

55.00

18.30

33.26

776.40

7.72

±SD

1.95

1.09

1.34

4.49

3.09

1.14

0.53

330.86

0.52

n

5

5

5

5

5

5

5

5

5

G2, M & 111

Mean

9.68

9.37

16.82

51.32

54.72

17.94

32.80

1135.20

7.46

±SD

2.59

0.15

0.33

0.89

1.12

0.29

0.58

184.30

0.19

n

5

5

5

5

5

5

5

5

5

G3, M & 333

Mean

8.67

9.40

16.58

50.78

54.12

17.64

32.62

962.60

7.72

±SD

3.67

0.62

0.87

1.91

1.72

0.59

0.70

256.00

0.52

n

5

5

5

5

5

5

5

5

5

G4, M & 1000

Mean

8.14

9.35

16.88

52.02

55.74

18.06

32.46

1017.60

7.50

±SD

2.42

0.54

0.55

1.58

2.71

0.74

0.53

93.02

0.38

n

5

5

5

5

5

5

5

5

5

M: Male; SD: Standard Deviation; n: Number of Animals (randomly selected).

TABLE 16 (Contd…). SUMMARY OF HAEMATOLOGY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Reticulocyte Count

Neutrophils

Lymphocytes

Monocytes

Eosinophils

Basophils

(Retic)

(Neut)

(Lymph)

(Mono)

(Eos)

(Baso)

(%)

(%)

(%)

(%)

(%)

(%)

G1, M & 0

Mean

2.25

23.48

72.02

2.38

0.44

0.80

±SD

1.24

9.09

9.80

1.15

0.17

0.49

n

5

5

5

5

5

5

G2, M & 111

Mean

1.87

19.66

75.74

2.28

1.04

0.54

±SD

0.64

3.44

3.55

0.74

0.59

0.23

n

5

5

5

5

5

5

G3, M & 333

Mean

1.87

21.98

70.58

2.40

1.04

0.62

±SD

0.60

5.35

7.57

2.37

0.34

0.61

n

5

5

5

5

5

5

G4, M & 1000

Mean

2.01

20.86

75.78

1.42

0.72

0.54

±SD

0.79

3.34

2.92

0.23

0.25

0.15

n

5

5

5

5

5

5

M: Male; SD: Standard Deviation; n: Number of Animals (randomly selected).

TABLE 16 (Contd…). SUMMARY OF HAEMATOLOGY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Absolute Reticulocyte Count

Absolute Neutrophils

Absolute Lymphocytes

Absolute Monocytes

Absolute Eosinophils

Absolute Basophils

Prothrombin Time

Activated Prothrombin Time

(Retic)

(Neut)

(Lymph)

(Mono)

(Eos)

(Baso)

(PT)

(APTT)

(109cells/L)

(103cells/µL)

(103cells/µL)

(103cells/µL)

(103cells/µL)

(103cells/µL)

(Seconds)

(Seconds)

G1, M & 0

Mean

212.96

1.83

6.07

0.19

0.04

0.07

15.54

22.26

±SD

101.87

0.42

2.01

0.08

0.02

0.06

0.78

6.12

n

5

5

5

5

5

5

5

5

G2, M & 111

Mean

174.50

1.91

7.33

0.23

0.10*

0.05

15.72

25.28

±SD

56.93

0.64

1.96

0.10

0.04

0.02

1.03

1.56

n

5

5

5

5

5

5

5

5

G3, M & 333

Mean

175.20

1.80

5.98

0.27

0.09*

0.07

16.72

24.30

±SD

59.25

0.52

1.97

0.39

0.03

0.11

1.49

1.59

n

5

5

5

5

5

5

5

5

G4, M & 1000

Mean

184.78

1.70

6.17

0.11

0.06

0.04

15.98

26.78

±SD

67.09

0.55

1.89

0.04

0.02

0.02

0.89

1.29

n

5

5

5

5

5

5

5

5

M: Male; SD: Standard Deviation; n: Number of Animals (randomly selected); *: Statistically significant (P<0.05) change than the vehicle control group.

TABLE 16 (Contd…). SUMMARY OF HAEMATOLOGY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Total

Leucocyte

Count

Total

Erythrocyte

Count

Haemoglobin

Haematocrit

Mean

Corpuscular Volume

Mean

Corpuscular Haemoglobin

Mean

Corpuscular Haemoglobin Concentration

Platelet

Count

Mean

Platelet

Volume

(WBC)

(RBC)

(HGB)

(HCT)

(MCV)

(MCH)

(MCHC)

(PLT)

(MPV)

(103cells/µL)

(106cells/µL)

(g/dL)

(%)

(fL)

(pg)

(g/dL)

(103cells/µL)

(fL)

G1, F & 0

Mean

10.56

7.85

15.74

46.26

59.02

20.04

34.04

752.60

6.02

±SD

3.78

0.45

0.65

3.16

4.16

1.08

0.91

361.58

0.64

n

5

5

5

5

5

5

5

5

5

G2, F & 111

Mean

11.38

7.41

14.66

44.60

60.26

19.74

32.82

1130.00

6.72

±SD

3.45

0.68

1.58

3.35

1.37

0.42

1.34

672.21

0.54

n

5

5

5

5

5

5

5

5

5

G3, F & 333

Mean

9.62

7.95

15.56

46.34

58.34

19.60

33.60

1030.20

6.42

±SD

3.95

0.28

0.72

2.05

3.12

0.81

1.07

104.22

0.57

n

5

5

5

5

5

5

5

5

5

G4, F & 1000

Mean

12.38

7.39

14.68

43.46

59.06

19.98

33.80

1003.00

6.92

±SD

3.58

0.76

0.89

2.75

3.76

1.58

0.63

227.17

0.68

n

5

5

5

5

5

5

5

5

5

F: Female; SD: Standard Deviation; n: Number of Animals (randomly selected).

TABLE 16 (Contd…). SUMMARY OF HAEMATOLOGY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Reticulocyte Count

Neutrophils

Lymphocytes

Monocytes

Eosinophils

Basophils

(Retic)

(Neut)

(Lymph)

(Mono)

(Eos)

(Baso)

(%)

(%)

(%)

(%)

(%)

(%)

G1, F & 0

Mean

2.28

32.72

60.66

3.68

1.46

0.60

±SD

0.87

15.77

16.88

1.06

0.98

0.23

n

5

5

5

5

5

5

G2, F & 111

Mean

4.52

30.00

62.56

4.60

1.18

0.42

±SD

4.56

11.00

11.24

1.35

0.63

0.19

n

5

5

5

5

5

5

G3, F & 333

Mean

1.49

25.26

66.92

4.46

1.22

0.80

±SD

0.67

9.73

12.89

4.23

0.33

1.12

n

5

5

5

5

5

5

G4, F & 1000

Mean

4.16

37.00

57.76

3.10

0.88

0.28

±SD

2.72

20.56

20.80

0.97

0.47

0.11

n

5

5

5

5

5

5

F: Female; SD: Standard Deviation; n: Number of Animals (randomly selected).

TABLE 16 (Contd…). SUMMARY OF HAEMATOLOGY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

 

Absolute Reticulocyte

Count  

Absolute

Neutrophils   

Absolute

Lymphocytes  

Absolute

Monocytes

Absolute

Eosinophils

Absolute

 Basophils       

Prothrombin

Time  

Activated Prothrombin

Time       

(Retic)

(Neut)

(Lymph)

(Mono)

(Eos)

(Baso)

(PT)

(APTT)

(109cells/L)

(103cells/µL)

(103cells/µL)

(103cells/µL)

(103cells/µL)

(103cells/µL)

(Seconds)

(Seconds)

G1, F & 0

Mean

177.36

3.64

6.21

0.40

0.15

0.06

17.60

23.34

±SD

60.94

2.11

2.30

0.19

0.09

0.03

1.02

2.05

n

5

5

5

5

5

5

5

5

G2, F & 111

Mean

313.86

3.59

6.91

0.56

0.12

0.05

22.06

27.48

±SD

276.22

2.17

1.72

0.29

0.04

0.02

8.41

10.47

n

5

5

5

5

5

5

5

5

G3, F & 333

Mean

117.60

2.64

6.29

0.39

0.12

0.06

16.88

22.08

±SD

50.69

2.06

2.26

0.31

0.07

0.08

3.84

7.81

n

5

5

5

5

5

5

5

5

G4, F & 1000

Mean

291.70

4.81

6.94

0.38

0.10

0.04

15.66

19.92

±SD

145.98

3.66

3.06

0.17

0.03

0.02

3.85

5.41

n

5

5

5

5

5

5

5

5

F: Female; SD: Standard Deviation; n: Number of Animals (randomly selected).

TABLE 16 (Contd…). SUMMARY OF HAEMATOLOGY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Total

Leucocyte

Count

Total

Erythrocyte

Count

Haemoglobin

Haematocrit

Mean

Corpuscular Volume

Mean

Corpuscular Haemoglobin

Mean

Corpuscular Haemoglobin Concentration

Platelet

Count

Mean

Platelet

Volume

(WBC)

(RBC)

(HGB)

(HCT)

(MCV)

(MCH)

(MCHC)

(PLT)

(MPV)

(103cells/µL)

(106cells/µL)

(g/dL)

(%)

(fL)

(pg)

(g/dL)

(103cells/µL)

(fL)

G1R, M & 0

Mean

9.13

9.21

16.04

48.76

52.94

17.42

32.92

894.80

6.50

±SD

0.78

0.14

0.43

1.79

1.78

0.39

0.42

134.10

0.31

n

5

5

5

5

5

5

5

5

5

G4R, M & 1000

Mean

9.55

9.14

15.80

48.54

53.08

17.30

32.62

1071.80

6.42

±SD

1.05

0.32

0.65

2.25

0.91

0.30

0.57

268.87

0.37

n

5

5

5

5

5

5

5

5

5

G1R, F & 0

Mean

6.97

7.65

14.26

42.16

55.20

18.64

33.78

873.60

6.32

±SD

2.14

0.43

0.65

2.05

1.99

0.70

0.29

80.79

0.19

n

5

5

5

5

5

5

5

5

5

G4R, F & 1000

Mean

7.59

7.73

14.32

42.78

55.34

18.48

33.46

739.60

6.52

±SD

3.51

0.22

0.57

1.63

2.15

0.70

0.25

253.21

0.13

n

5

5

5

5

5

5

5

5

5

M: Male; F: Female; R: Recovery; SD: Standard Deviation; n: Number of Animals.

TABLE 16 (Contd…). SUMMARY OF HAEMATOLOGY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Reticulocyte Count

Neutrophils

Lymphocytes

Monocytes

Eosinophils

Basophils

(Retic)

(Neut)

(Lymph)

(Mono)

(Eos)

(Baso)

(%)

(%)

(%)

(%)

(%)

(%)

G1R, M & 0

Mean

1.63

25.28

69.00

1.76

2.30

0.88

±SD

0.26

6.62

7.82

0.49

0.89

0.26

n

5

5

5

5

5

5

G4R, M & 1000

Mean

2.03

21.70

73.32

1.48

2.02

0.60

±SD

0.50

5.52

5.80

0.37

0.36

0.29

n

5

5

5

5

5

5

G1R, F & 0

Mean

1.84

26.12

68.68

2.30

1.34

0.68

±SD

0.36

9.06

8.57

1.19

0.45

0.15

n

5

5

5

5

5

5

G4R, F & 1000

Mean

2.48*

29.02

64.16

2.40

2.86*

0.64

±SD

0.39

7.89

6.96

0.87

1.30

0.13

n

5

5

5

5

5

5

M: Male; F: Female; R: Recovery; SD: Standard Deviation; n: Number of Animals; *: Statistically significant (P<0.05) change than the vehicle control group.

TABLE 16 (Contd…). SUMMARY OF HAEMATOLOGY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

 

Absolute

Reticulocyte Count  

Absolute

Neutrophils   

Absolute

Lymphocytes  

Absolute

Monocytes

Absolute

Eosinophils

Absolute 

Basophils       

Prothrombin

Time  

Activated

Prothrombin Time       

(Retic)

(Neut)

(Lymph)

(Mono)

(Eos)

(Baso)

(PT)

(APTT)

(109cells/L)

(103cells/µL)

(103cells/µL)

(103cells/µL)

(103cells/µL)

(103cells/µL)

(Seconds)

(Seconds)

G1R, M & 0

Mean

150.40

2.27

6.35

0.16

0.21

0.08

16.00

21.56

±SD

23.36

0.40

1.23

0.04

0.07

0.02

2.04

2.05

n

5

5

5

5

5

5

5

5

G4R, M & 1000

Mean

186.92

2.06

7.02

0.14

0.19

0.05

15.50

22.48

±SD

52.68

0.53

1.09

0.04

0.04

0.03

1.17

3.12

n

5

5

5

5

5

5

5

5

G1R, F & 0

Mean

140.08

1.78

4.83

0.15

0.09

0.05

14.98

24.56

±SD

23.05

0.73

1.73

0.05

0.02

0.01

0.53

1.98

n

5

5

5

5

5

5

5

5

G4R, F & 1000

Mean

192.10*

2.06

4.98

0.17

0.25

0.05

15.92

19.34*

±SD

30.17

0.60

2.77

0.07

0.24

0.02

1.17

2.34

n

5

5

5

5

5

5

5

5

M: Male; F: Female; R: Recovery; SD: Standard Deviation; n: Number of Animals; *: Statistically significant (P<0.05) change than the vehicle control group.

TABLE 17.     SUMMARY OF CLINICAL CHEMISTRY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Glucose

Urea

Creatinine

Total Cholesterol

Triglycerides

Total Protein

Albumin

(GLU)

-

(CRE)

(CHO)

(TRI)

(TPR)

(ALB)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

(g/dL)

(g/dL)

G1, M & 0

Mean

126.60

30.52

0.58

51.60

25.60

7.74

3.47

±SD

15.47

2.56

0.03

3.36

2.30

0.26

0.19

n

5

5

5

5

5

5

5

G2, M & 111

Mean

123.60

33.30

0.57

60.60

30.40

7.24*

3.55

±SD

5.55

3.91

0.04

4.83

6.07

0.33

0.15

n

5

5

5

5

5

5

5

G3, M & 333

Mean

122.60

33.78

0.60

58.00

29.40

7.44

3.65

±SD

12.92

2.83

0.03

6.71

7.73

0.22

0.29

n

5

5

5

5

5

5

5

G4, M & 1000

Mean

131.00

36.04

0.59

56.00

27.00

7.58

3.68

±SD

19.79

4.26

0.02

9.57

6.16

0.29

0.24

n

5

5

5

5

5

5

5

M: Male; SD: Standard Deviation; n: Number of Animals (randomly selected); *: Statistically significant (P<0.05) change than the vehicle control group.

TABLE 17 (Contd…). SUMMARY OF CLINICAL CHEMISTRY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Alanine
aminotransferase

Aspartate
aminotransferase

Alkaline
phosphatase

Total Bilirubin

Calcium

Phosphorous

(ALT)

(AST)

(ALP)

(BIT)

(CAL)

(PHO)

(U/L)

(U/L)

(U/L)

(mg/dL)

(mg/dL)

(mg/dL)

G1, M & 0

Mean

61.20

111.20

125.80

0.05

9.36

6.48

±SD

16.93

12.34

21.11

0.01

0.11

0.58

n

5

5

5

5

5

5

G2, M & 111

Mean

47.40

93.20

120.00

0.06

9.64

6.54

±SD

8.96

13.46

31.92

0.03

0.47

0.62

n

5

5

5

5

5

5

G3, M & 333

Mean

40.00

81.60*

127.20

0.06

10.40*

6.82

±SD

5.96

12.30

38.01

0.03

1.05

0.62

n

5

5

5

5

5

5

G4, M & 1000

Mean

52.20

91.80

114.80

0.06

9.94

6.62

±SD

24.52

18.69

28.70

0.02

0.30

0.49

n

5

5

5

5

5

5

M: Male; SD: Standard Deviation; n: Number of Animals (randomly selected) ; *: Statistically significant (P<0.05) change than the vehicle control group.

TABLE 17 (Contd…). SUMMARY OF CLINICAL CHEMISTRY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Globulin

Albumin/

Globulin ratio

Blood Urea Nitrogen

Sodium

Potassium

Chloride

(GLO)

(A/G Ratio)

(BUN)

(Na)

(K)

(CLO)

(g/dL)

-

(mg/dL)

(mmol/L)

(mmol/L)

(mmol/L)

G1, M & 0

Mean

4.27

0.82

14.24

141.64

3.96

104.10

±SD

0.30

0.09

1.19

0.94

0.25

2.51

n

5

5

5

5

5

5

G2, M & 111

Mean

3.69*

0.97

15.54

142.16

3.71

105.38

±SD

0.38

0.12

1.83

0.53

0.15

1.40

n

5

5

5

5

5

5

G3, M & 333

Mean

3.79*

0.97

15.77

142.44

3.86

105.78

±SD

0.12

0.10

1.32

0.93

0.36

1.73

n

5

5

5

5

5

5

G4, M & 1000

Mean

3.90

0.95

16.82

142.76

3.85

106.40

±SD

0.29

0.11

1.99

0.66

0.09

0.86

n

5

5

5

5

5

5

M: Male; SD: Standard Deviation; n: Number of Animals (randomly selected); *: Statistically significant (P<0.05) change than the vehicle control group.

TABLE 17 (Contd…). SUMMARY OF CLINICAL CHEMISTRY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Glucose

Urea

Creatinine

Total Cholesterol

Triglycerides

Total Protein

Albumin

(GLU)

-

(CRE)

(CHO)

(TRI)

(TPR)

(ALB)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

(g/dL)

(g/dL)

G1, F & 0

Mean

102.20

48.68

0.54

76.60

78.60

7.32

3.63

±SD

26.22

8.11

0.05

22.67

26.83

1.08

0.67

n

5

5

5

5

5

5

5

G2, F & 111

Mean

121.20

51.14

0.52

67.80

59.40

7.34

3.41

±SD

29.64

13.28

0.03

17.67

23.65

0.75

0.30

n

5

5

5

5

5

5

5

G3, F & 333

Mean

103.80

44.78

0.57

75.60

43.20

7.56

3.58

±SD

13.81

19.81

0.04

14.91

21.48

0.29

0.32

n

5

5

5

5

5

5

5

G4, F & 1000

Mean

114.60

51.66

0.57

75.80

105.20

7.76

3.70

±SD

16.44

16.16

0.07

20.44

68.28

0.58

0.11

n

5

5

5

5

5

5

5

F: Female; SD: Standard Deviation; n: Number of Animals (randomly selected).

TABLE 17 (Contd…). SUMMARY OF CLINICAL CHEMISTRY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Alanine
aminotransferase

Aspartate
aminotransferase

Alkaline
phosphatase

Total Bilirubin

Calcium

Phosphorous

(ALT)

(AST)

(ALP)

(BIT)

(CAL)

(PHO)

(U/L)

(U/L)

(U/L)

(mg/dL)

(mg/dL)

(mg/dL)

G1, F & 0

Mean

84.60

119.40

164.40

0.03

10.06

5.92

±SD

14.43

14.31

75.62

0.04

1.06

0.95

n

5

5

5

5

5

5

G2, F & 111

Mean

113.80

143.80

420.00

0.02

9.42

5.50

±SD

66.08

71.78

367.29

0.02

0.64

1.64

n

5

5

5

5

5

5

G3, F & 333

Mean

96.20

99.40

305.80

0.06

10.04

6.06

±SD

82.35

16.01

412.83

0.02

0.85

1.93

n

5

5

5

5

5

5

G4, F & 1000

Mean

88.60

101.80

212.40

0.03

10.38

6.38

±SD

46.69

22.65

81.17

0.04

0.91

3.11

n

5

5

5

5

5

5

F: Female; SD: Standard Deviation; n: Number of Animals (randomly selected).

TABLE 17 (Contd…). SUMMARY OF CLINICAL CHEMISTRY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Globulin

Albumin/Globulin ratio

Blood Urea Nitrogen

Sodium

Potassium

Chloride

(GLO)

(A/G Ratio)

(BUN)

(Na)

(K)

(CLO)

(g/dL)

-

(mg/dL)

(mmol/L)

(mmol/L)

(mmol/L)

G1, F & 0

Mean

3.69

0.98

22.72

143.04

4.02

106.18

±SD

0.50

0.11

3.79

1.34

0.81

2.96

n

5

5

5

5

5

5

G2, F & 111

Mean

3.93

0.88

23.87

142.18

3.57

107.22

±SD

0.66

0.14

6.20

1.47

0.28

3.01

n

5

5

5

5

5

5

G3, F & 333

Mean

3.98

0.91

20.90

142.36

3.60

107.94

±SD

0.40

0.16

9.25

1.61

0.33

2.63

n

5

5

5

5

5

5

G4, F & 1000

Mean

4.06

0.92

24.11

142.62

3.86

107.06

±SD

0.50

0.09

7.55

1.62

0.40

3.20

n

5

5

5

5

5

5

F: Female; SD: Standard Deviation; n: Number of Animals (randomly selected).

TABLE 17 (Contd…). SUMMARY OF CLINICAL CHEMISTRY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Glucose

Urea

Creatinine

Total Cholesterol

Triglycerides

Total Protein

Albumin

(GLU)

-

(CRE)

(CHO)

(TRI)

(TPR)

(ALB)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

(g/dL)

(g/dL)

G1R, M & 0

Mean

99.80

27.36

0.55

59.60

23.00

6.98

3.10

±SD

6.53

3.92

0.02

16.21

5.52

0.26

0.07

n

5

5

5

5

5

5

5

G4R, M & 1000

Mean

99.40

25.74

0.56

68.80

31.60

7.12

3.05

±SD

10.53

1.19

0.03

8.76

10.78

0.11

0.04

n

5

5

5

5

5

5

5

G1R, F & 0

Mean

120.40

35.68

0.61

106.40

49.60

8.44

4.10

±SD

7.16

7.57

0.02

17.07

34.09

0.56

0.18

n

5

5

5

5

5

5

5

G4R, F & 1000

Mean

104.00*

31.42

0.59

89.20

41.60

8.04

3.70

±SD

13.49

2.25

0.01

16.98

11.87

0.42

0.44

n

5

5

5

5

5

5

5

M: Male; R: Recovery; SD: Standard Deviation; n: Number of Animals; *: Statistically significant (P<0.05) change than the vehicle control group.

TABLE 17 (Contd…). SUMMARY OF CLINICAL CHEMISTRY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Alanine aminotransferase

Aspartate aminotransferase

Alkaline phosphatase

Total Bilirubin

Calcium

Phosphorous

(ALT)

(AST)

(ALP)

(BIT)

(CAL)

(PHO)

(U/L)

(U/L)

(U/L)

(mg/dL)

(mg/dL)

(mg/dL)

G1R, M & 0

Mean

52.00

101.80

99.00

0.02

10.42

5.72

±SD

40.34

53.17

23.40

0.02

0.37

0.34

n

5

5

5

5

5

5

G4R, M & 1000

Mean

42.60

98.40

105.20

0.02

10.02

5.26

±SD

9.86

18.49

29.84

0.01

0.15

0.50

n

5

5

5

5

5

5

G1R, F & 0

Mean

54.00

94.00

70.80

0.02

11.20

4.20

±SD

28.82

36.89

40.92

0.02

0.45

0.38

n

5

5

5

5

5

5

G4R, F & 1000

Mean

53.80

103.20

69.00

0.04

10.90

4.42

±SD

21.53

39.30

60.10

0.02

0.37

0.18

n

5

5

5

5

5

5

M: Male; R: Recovery; SD: Standard Deviation; n: Number of Animals.

TABLE 17 (Contd…). SUMMARY OF CLINICAL CHEMISTRY RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Globulin

Albumin/

Globulin ratio

Blood Urea Nitrogen

Sodium

Potassium

Chloride

(GLO)

(A/G Ratio)

(BUN)

(Na)

(K)

(CLO)

(g/dL)

-

(mg/dL)

(mmol/L)

(mmol/L)

(mmol/L)

G1R, M & 0

Mean

3.88

0.80

12.77

142.82

3.56

105.48

±SD

0.27

0.06

1.83

0.36

0.41

0.92

n

5

5

5

5

5

5

G4R, M & 1000

Mean

4.07

0.75

12.01

143.16

3.59

108.06*

±SD

0.15

0.04

0.56

0.68

0.41

1.65

n

5

5

5

5

5

5

G1R, F & 0

Mean

4.34

0.95

16.65

142.62

3.45

105.76

±SD

0.42

0.08

3.53

0.94

0.17

1.52

n

5

5

5

5

5

5

G4R, F & 1000

Mean

4.34

0.85

14.66

142.26

3.62

106.40

±SD

0.21

0.12

1.05

1.41

0.48

2.88

n

5

5

5

5

5

5

M: Male; R: Recovery; SD: Standard Deviation; n: Number of Animals; *: Statistically significant (P<0.05) change than the vehicle control group.

TABLE 18.     SUMMARY OF URINALYSIS RECORD

Examination

Group, Sex & Dose

(mg/kg body weight/day)

G1, M & 0

G2, M & 111

G3, M & 333

G4, M & 1000

Number of Animals

(randomly selected)

5

5

5

5

Physical Examination

Color

Pale Yellow

5

5

5

5

Appearance

Clear

5

5

5

5

Volume (mL)

Mean

10.6

10.2

11.0

11.4

±SD

0.7

1.4

0.8

0.4

Chemical Examination

pH

Mean

7.5

7.4

7.4

7.5

±SD

0.0

0.4

0.5

0.0

Specific Gravity

Mean

1.008

1.011

1.009

1.009

±SD

0.003

0.002

0.002

0.002

Urobilinogen (mg/dL)

Mean

0.2

0.2

0.2

0.2

±SD

0.0

0.0

0.0

0.0

Bilirubin (mg/dL)

Neg

5

5

5

5

Ketones (mg/dL)

Neg

2

-

2

1

5

3

5

3

4

Blood

(Ery/µL)

Neg

5

4

5

5

Ca80

-

1

-

-

Protein

(mg/dL)

Neg

3

-

3

1

Trace

2

4

1

4

30

-

1

1

-

Nitrite

Neg

4

4

5

1

Pos

1

1

-

4

Leucocytes

Neg

3

-

3

1

Ca15

1

1

1

2

Ca70

1

3

1

2

Ca125

-

1

-

-

Glucose

(mg/dL)

Neg

5

5

5

5

Microalbumin

(mg/dL)

Neg

-

-

2

1

>15

2

5

2

4

15

3

-

1

-

Microscopic

Examination

Epi Cells

0

3

4

2

4

0-1

1

1

2

1

1-2

1

-

1

-

Casts

Absent

4

4

4

5

Present

1

1

1

-

Crystals

Present

5

5

5

5

M: Male; SD: Standard Deviation; Pos: Positive; Neg: Negative; Ca: Calculated.
 >=: Greater than or equal to; >: Greater than.

TABLE 18 (Contd…). SUMMARY OF URINALYSIS RECORD

Examination

Group, Sex & Dose

(mg/kg body weight/day)

G1R,

M & 0

G4R,

M &1000

G1R,

F & 0

G4R,

F & 1000

Number of Animals

(randomly selected)

5

5

5

5

Physical Examination

Color

Pale Yellow

2

5

3

3

Yellow

3

-

2

2

Appearance

Clear

3

2

4

4

Turbid

2

3

1

1

Volume (mL)

Mean

6.4

8.6

7.9

7.8

±SD

1.7

3.1

2.2

0.8

Chemical Examination

pH

Mean

6.7

7.3

6.8

7.6

±SD

0.9

0.4

1.0

1.0

Specific Gravity

Mean

1.020

1.014

1.014

1.012

±SD

0.007

0.002

0.007

0.008

Urobilinogen (mg/dL)

Mean

0.2

0.2

0.2

0.6

±SD

0.0

0.0

0.0

0.8

Bilirubin (mg/dL)

Neg

2

5

5

2

1

3

-

-

3

Ketones (mg/dL)

Neg

1

2

5

5

5

4

3

-

-

Blood

(Ery/µL)

Neg

5

5

5

4

Ca10

-

-

-

1

Protein

(mg/dL)

Neg

2

5

4

2

Trace

2

-

1

-

30

1

-

-

-

100

-

-

-

1

>=300

-

-

-

2

Nitrite

Neg

3

5

5

5

Pos

2

-

-

-

Leucocytes

Neg

-

1

3

2

Ca15

1

4

2

-

Ca70

4

-

-

3

Glucose

(mg/dL)

Neg

5

5

5

5

Microalbumin

(mg/dL)

Neg

-

1

3

1

>15

3

-

1

3

15

2

4

1

1

Microscopic Examination\

Epi Cells

0

3

3

2

2

0-1

1

2

2

1

1-2

1

-

1

-

2-3

-

-

-

2

Casts

Absent

5

2

5

4

Present

-

3

-

1

Crystals

Absent

-

2

1

2

Present

5

3

4

3

M: Male; F: Female; R: Recovery; SD: Standard Deviation; Pos: Positive; Neg: Negative.                              
Ca: Calculated; >=: Greater than or equal to; >: Greater than.

TABLE 19.     SUMMARY OF VAGINAL SMEAR EXAMINATION RECORD FOR DETERMINATION OF OESTRUS CYCLICITY

Group, Sex

& Dose
(mg/kg body weight/day)

Total No. of

Females Evaluated

Females with

Complete

Regular Cycles

Females with at least one Irregular

Oestrus Cycle

Average Length of

Oestrus Cycle (Days)

G1, F & 0

12

n

11

1

Mean

4.65

%

91.7

8.3

±SD

0.41

n

12

G2, F & 111

12

n

11

Mean

4.43

%

91.7

0.0

±SD

0.42

n

12

G3, F & 333

12

n

12

Mean

4.68

%

100.0

0.0

±SD

0.27

n

12

G4, F & 1000

12

n

11

1

Mean

4.53

%

91.7

8.3

±SD

0.34

n

12

F: Female; SD: Standard Deviation; n: Number of Animals.

TABLE 20.     SUMMARY OF DELIVERY AND LITTER OBSERVATIONS RECORD

Group, Sex

& Dose
(mg/kg body weight/day)

Delivery Record At birth

Litter

Size

(No.)

Live Pups (No.)

Dead Pups (No.)

Cannibalized Pups (No.)

Sex Ratio (m/f)

at Birth

Live Birth Index

(%)

Male

Female

Total

Male

Female

Total

Undetermined

Male

Female

Total

G1, F & 0

Mean

12.18

6.27

5.73

12.00

0.18

0.00

0.18

0.00

0.00

0.00

0.00

 

1.14

98.86

±SD

2.68

1.90

1.35

2.41

0.40

0.00

0.40

0.00

0.00

0.00

0.00

0.42

2.54

n

11

11

11

11

11

11

11

11

11

11

11

11

11

G2, F & 111

Mean

12.20

 

6.00

6.00

12.00

 

0.10

0.10

0.20

 

0.00

0.00

0.00

0.00

 

1.12

 

98.70

±SD

3.29

1.94

1.89

3.09

0.32

0.32

0.42

0.00

0.00

0.00

0.00

0.57

2.76

n

10

10

10

10

10

10

10

10

10

10

10

10

10

G3, F & 333

Mean

10.90

 

5.30

5.50

10.80

 

0.10

0.00

0.10

 

0.00

0.00

0.00

0.00

 

1.00

 

99.41

±SD

2.77

2.00

1.27

2.53

0.32

0.00

0.32

0.00

0.00

0.00

0.00

0.39

1.86

n

10

10

10

10

10

10

10

10

10

10

10

10

10

G4, F & 1000

Mean

12.82

 

6.36

6.09

12.45

 

0.18

0.18

0.36

 

0.00

0.00

0.00

0.00

 

1.16

 

96.57

±SD

1.99

2.11

1.64

2.54

0.40

0.60

0.92

0.00

0.00

0.00

0.00

0.71

9.11

n

11

11

11

11

11

11

11

11

11

11

11

11

11

F: Female; SD: Standard Deviation; n: Number of Dams; m/f: Male/Female; %: Percentage.

TABLE 20 (Contd…). SUMMARY RECORD OF DELIVERY AND LITTER OBSERVATIONS

Group, Sex

& Dose
(mg/kg body

weight/day)

LD 1 to 4

Sex Ratio

(m/f)

on LD 4

Pup Survival

Index (%)

[LD 1 to 4]

Live Pups (No.)

Dead Pups (No.)

Cannibalized Pups (No.)

Male

Female

Total

Male

Female

Total

Male

Female

Total

G1, F & 0

Mean

6.27

5.73

12.00

0.00

0.00

0.00

0.00

0.00

0.00

1.14

100.00

±SD

1.90

1.35

2.41

0.00

0.00

0.00

0.00

0.00

0.00

0.42

0.00

n

11

11

11

11

11

11

11

11

11

11

11

G2, F & 111

Mean

6.00

6.00

12.00

0.00

0.00

0.00

0.00

0.00

0.00

1.12

100.00

±SD

1.94

1.89

3.09

0.00

0.00

0.00

0.00

0.00

0.00

0.57

0.00

n

10

10

10

10

10

10

10

10

10

10

10

G3, F & 333

Mean

5.30

5.50

10.80

0.00

0.00

0.00

0.00

0.00

0.00

1.00

100.00

±SD

2.00

1.27

2.53

0.00

0.00

0.00

0.00

0.00

0.00

0.39

0.00

n

10

10

10

10

10

10

10

10

10

10

10

G4, F & 1000

Mean

6.36

6.09

12.45

0.00

0.00

0.00

0.00

0.00

0.00

1.16

100.00

±SD

2.11

1.64

2.54

0.00

0.00

0.00

0.00

0.00

0.00

0.71

0.00

n

11

11

11

11

11

11

11

11

11

11

11

F: Female; SD: Standard Deviation; n: Number of Dams; m/f: Male/Female; %: Percentage.

TABLE 20 (Contd…). SUMMARY RECORD OF DELIVERY AND LITTER OBSERVATIONS

Group, Sex

& Dose
(mg/kg body weight/day)

Pups Sacrificed for Litter Standardization /

Hormonal analysis (No.)

Live Pups (No.) on LD 4 after Litter Standardization / Hormonal analysis (No.)

LD 5 to 7

Sex Ratio (m/f)

on LD 7

Pup Survival Index (%)

[LD 5 to 7]

Live Pups (No.)

Dead Pups (No.)

Cannibalized (No.)

Male

Female

Total

Male

Female

Total

Male

Female

Total

Male

Female

Total

Male

Female

Total

G1, F & 0

Mean

0.00

1.09

1.09

6.27

4.64

10.91

6.27

4.64

10.91

0.00

0.00

0.00

0.00

0.00

0.00

1.55

100.00

±SD

0.00

1.04

1.04

1.90

1.36

1.58

1.90

1.36

1.58

0.00

0.00

0.00

0.00

0.00

0.00

0.86

0.00

n

11

11

11

11

11

11

11

11

11

11

11

11

11

11

11

11

11

G2, F & 111

Mean

0.00

1.20

1.20

6.00

4.80

10.80

6.00

4.80

10.80

0.00

0.00

0.00

0.00

0.00

0.00

1.37

100.00

±SD

0.00

1.03

1.03

1.94

1.23

2.20

1.94

1.23

2.20

0.00

0.00

0.00

0.00

0.00

0.00

0.62

0.00

n

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

G3, F & 333

Mean

0.00

0.70

0.70

5.30

4.80

10.10

5.30

4.80

10.10

0.00

0.00

0.00

0.00

0.00

0.00

1.21

100.00

±SD

0.00

0.95

0.95

2.00

1.03

1.73

2.00

1.03

1.73

0.00

0.00

0.00

0.00

0.00

0.00

0.67

0.00

n

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

10

G4, F & 1000

Mean

0.00

1.64

1.64

6.36

4.45

10.82

6.36

4.45

10.82

0.00

0.00

0.00

0.00

0.00

0.00

2.07

100.00

±SD

0.00

0.81

0.81

2.11

1.57

1.99

2.11

1.57

1.99

0.00

0.00

0.00

0.00

0.00

0.00

2.39

0.00

n

11

11

11

11

11

11

11

11

11

11

11

11

11

11

11

11

11

F: Female; SD: Standard Deviation; n: Number of Dams; LD: Lactation Day; m/f: Male/Female.

TABLE 20 (Contd…). SUMMARY RECORD OF DELIVERY AND LITTER OBSERVATIONS

Group, Sex

& Dose
(mg/kg body

weight/day)

LD 8 to 13

Sex Ratio

(m/f)

on LD 13

Pup Survival

Index (%)

[LD 8 to 13]

Live Pups (No.)

Dead Pups (No.)

Cannibalized (No.)

Male

Female

Total

Male

Female

Total

Male

Female

Total

G1, F & 0

Mean

6.27

4.64

10.91

0.00

0.00

0.00

0.00

0.00

0.00

1.55

100.00

±SD

1.90

1.36

1.58

0.00

0.00

0.00

0.00

0.00

0.00

0.86

0.00

n

11

11

11

11

11

11

11

11

11

11

11

G2, F & 111

Mean

6.00

4.80

10.80

0.00

0.00

0.00

0.00

0.00

0.00

1.37

100.00

±SD

1.94

1.23

2.20

0.00

0.00

0.00

0.00

0.00

0.00

0.62

0.00

n

10

10

10

10

10

10

10

10

10

10

10

G3, F & 333

Mean

5.30

4.80

10.10

0.00

0.00

0.00

0.00

0.00

0.00

1.21

100.00

±SD

2.00

1.03

1.73

0.00

0.00

0.00

0.00

0.00

0.00

0.67

0.00

n

10

10

10

10

10

10

10

10

10

10

10

G4, F & 1000

Mean

6.36

4.45

10.82

0.00

0.00

0.00

0.00

0.00

0.00

2.07

100.00

±SD

2.11

1.57

1.99

0.00

0.00

0.00

0.00

0.00

0.00

2.39

0.00

n

11

11

11

11

11

11

11

11

11

11

11

F: Female; SD: Standard Deviation; n: Number of Dams; LD: Lactation Day; m/f: Male/Female; %: Percentage.

TABLE 21.     SUMMARY RECORD OF REPRODUCTIVE PERFORMANCE

Group, Sex

& Dose  
(mg/kg body weight/day)

No. of Males with Evidence of Mating
(No. of Males used for Mating)

Male Mating Index (%)

No. of Males Capable of Impregnating a Female
(No. of males used for Mating)

Male Fertility Index (%)

G1, M & 0

12 (12)

100.0

11 (12)

91.7

G2, M & 111

12 (12)

100.0

10 (12)

83.3

G3, M & 333

12 (12)

100.0

10 (12)

83.3

G4, M & 1000

12 (12)

100.0

11 (12)

91.7

M: Male.

TABLE 21 (Contd…). SUMMARY RECORD OF REPRODUCTIVE PERFORMANCE

Group, Sex

& Dose

(mg/kg body

weight/day)

Cohabitation Record and Pre-coital Interval (Mean Time of Mating)

 

Gestational Length

(duration of pregnancy)

Pre-coital Interval (Days)

Conceiving Days

(1 to 5)

Conceiving Days

(More than 5 days)

Gestation Length (Days)

G1, F & 0

Mean

3.58

n

8

4

22.55

SD

3.80

%

66.67

33.33

0.52

N

12

11

G2, F & 111

Mean

3.75

n

9

3

22.20

SD

4.45

%

75

25

0.63

N

12

10

G3, F & 333

Mean

4.58

n

8

4

22.50

SD

5.00

%

66.67

33.33

0.53

N

12

10

G4, F & 1000

Mean

4.67

n

8

4

22.36

SD

5.50

%

66.67

33.33

0.50

N

12

11

F: Female; SD: Standard Deviation; n: Number of females with evidence of mating (pre-coital interval); n: Number of pregnant females (gestation length).

TABLE 21 (Contd…). SUMMARY RECORD OF REPRODUCTIVE PERFORMANCE

Group, Sex

& Dose

(mg/kg body

weight/day)

Female Mating Index (%)

Female Fertility Index (%)

No. of Females with Evidence of Mating

(No. of Females used for Mating)

Female Mating Index (%)

No. of Females Confirmed as Fertile
(No. of Females used for Mating)

Female Fertility Index (%)

G1, F & 0

12 (12)

100.0

11 (12)

91.7

G2, F & 111

12 (12)

100.0

10 (12)

83.3

G3, F & 333

12 (12)

100.0

10 (12)

83.3

G4, F & 1000

12 (12)

100.0

11 (12)

91.7

F: Female.

TABLE 21 (Contd…). SUMMARY RECORD OF REPRODUCTIVE PERFORMANCE

Group, Sex

& Dose

mg/kg body

weight/day)

Female Fecundity or Pregnancy Index (%)

Gestation Index (%)

No. of Pregnant Females

No. of Females

Confirmed with Mating

Female Fecundity or Pregnancy Index (%)

Females with Live Born

Pups at Parturition

No. of Females

with Evidence of Pregnancy

Gestation Index (%)

G1, F & 0

11

12

91.7

11

11

100.0

G2, F & 111

10

12

83.3

10

10

100.0

G3, F & 333

10

12

83.3

10

10

100.0

G4, F & 1000

11

12

91.7

11

11

100.0

F: Female.

TABLE 21 (Contd…). SUMMARY RECORD OF REPRODUCTIVE PERFORMANCE

Group, Sex & Dose

 (mg/kg body weight/day)

Parturition index (%)

No. of Females Littered

No. of Females with evidence of pregnancy

Parturition index (%)

G1, F & 0

11

11

100.0

G2, F & 111

10

10

100.0

G3, F & 333

10

10

100.0

G4, F & 1000

11

11

100.0

F: Female.

TABLE 21 (Contd…). SUMMARY RECORD OF REPRODUCTIVE PERFORMANCE

Group, Sex

& Dose
(mg/kg body

weight/day)

 

Post-implantation Loss (%)

 

Post-natal Loss (%)

No. of

Implantations

No. of Viable

(Live)

Pups

Post-implantation Loss

(No.)

Post-implantation Loss

(%)

Total No. of Pups Found Dead/

Cannibalized during lactation period

Post-natal Loss

(%)

G1, F & 0

Mean

12.45

12.00

0.45

3.70

 

0.00

0.00

±SD

2.42

2.41

0.52

4.40

 

0.00

0.00

n

11

11

11

11

 

11

11

G2, F & 111

Mean

12.70

12.00

0.70

6.02

 

0.00

0.00

±SD

2.75

3.09

1.06

9.67

 

0.00

0.00

n

10

10

10

10

 

10

10

G3, F & 333

Mean

11.00

10.80

0.20

1.50

 

0.00

0.00

±SD

2.75

2.53

0.42

3.25

 

0.00

0.00

n

10

10

10

10

 

10

10

G4, F & 1000

Mean

13.27

12.45

0.82

6.72

 

0.00

0.00

±SD

1.74

2.54

1.40

11.56

 

0.00

0.00

n

11

11

11

11

 

11

11

F: Female; n: Number of Dams.

TABLE 22.     SUMMARY OF ABSOLUTE ORGAN WEIGHT (g) RECORD

Group, Sex

& Dose

(mg/kg body weight/day)

Adrenals

Thymus

Spleen

Testes

Epididymides

Heart

Kidneys

Brain

G1, M & 0

Mean

0.0667

0.2891

0.7915

3.3315

1.4198

1.3943

2.9537

2.1193

±SD

0.0045

0.0466

0.2701

0.3035

0.1420

0.2048

0.2510

0.0321

n

5

5

5

12

12

5

5

5

G2, M & 111

Mean

0.0719

0.3589

0.8023

3.4284

1.4112

1.5033

2.8196

2.1336

±SD

0.0094

0.0292

0.0928

0.1782

0.1365

0.1061

0.1723

0.0957

n

5

5

5

12

12

5

5

5

G3, M & 333

Mean

0.0709

0.4450*

0.8041

3.3728

1.4195

1.4754

2.8317

2.1467

±SD

0.0060

0.0665

0.1455

0.2771

0.1776

0.1560

0.1507

0.0446

n

5

5

5

12

12

5

5

5

G4, M & 1000

Mean

0.0716

0.4129*

0.8155

3.3454

1.5221

1.5218

3.0082

2.2033

±SD

0.0115

0.1013

0.1404

0.2101

0.1812

0.3979

0.7565

0.1355

n

5

5

5

12

12

5

5

5

M: Male; SD: Standard Deviation; n: Number of Animals; *: Statistically significant (P<0.05) change than the vehicle control group.

TABLE 22 (Contd…). SUMMARY OF ABSOLUTE ORGAN WEIGHT (g) RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

 

Liver

Prostate

Seminal vesicles with

coagulating glands

Lungs

Pituitary

Thyroid along with

parathyroid#

G1, M & 0

Mean

10.0491

1.1499

1.6300

1.9467

0.0208

0.0296

±SD

1.6192

0.1975

0.4468

0.1258

0.0014

0.0017

n

5

12

12

5

5

12

G2, M & 111

Mean

10.3930

1.1988

1.6027

2.0659

0.0197

0.0290

±SD

0.4389

0.2552

0.3454

0.1175

0.0013

0.0020

n

5

12

12

5

5

12

G3, M & 333

Mean

10.6576

1.3430

1.7846

2.0478

0.0203

0.0301

±SD

0.7376

0.3501

0.3700

0.1024

0.0009

0.0013

n

5

12

12

5

5

12

G4, M &

1000

Mean

10.8065

1.2209

1.9124

2.0837

0.0205

0.0305

±SD

2.8155

0.3931

0.5906

0.1492

0.0015

0.0023

n

5

12

12

5

5

12

M: Male; SD: Standard Deviation; n: Number of Animals; #: Weighed post fixation.

TABLE 22 (Contd…). SUMMARY OF ABSOLUTE ORGAN WEIGHT (g) RECORD

Group, Sex

& Dose

(mg/kg body

weight/day)

Adrenals

Thymus

Spleen

Ovaries

Uterus with Cervix

Heart

G1, F & 0

Mean

0.0741

0.2781

0.7048

0.1738

0.5901

1.1848

±SD

0.0083

0.0317

0.0646

0.0299

0.1167

0.1262

n

5

5

5

12

11@

5

G2, F & 111

Mean

0.0884

0.2632

0.7503

0.1432*

0.5097

1.1940

±SD

0.0110

0.0345

0.1289

0.0332

0.1138

0.0729

n

5

5

5

12

11@

5

G3, F & 333

Mean

0.0728

0.2267*

0.6584

0.1291*

0.5413

1.2001

±SD