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Diss Factsheets

Administrative data

Description of key information

Oral:

14 -day dose range finding study for OECD 407 study: NOAEL: 1000 mg/kg bw

28 -day repeated dose toxicity study (OECD 407): NOAEL: 1000 mg/kg bw

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
05 april 2018 to 08 June 2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)
Version / remarks:
adopted on 03 October 2008
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Limit test:
no
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and batch No.of test material: DEG4 410913
- Expiration date of the batch: 2019-02-22

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient (21 to 29oC)
- Solubility and stability of the test substance in the vehicle: soluble in corn oil and The stability of the test item Hostacor ITD in dose formulations was established under Bioneeds Study No.: BIO-ANM 813. As per the stability results, the test item formulations at the concentrations of 1 mg/mL and 100 mg/mL are stable up to 6 hours at room temperature




Species:
rat
Strain:
Sprague-Dawley
Details on species / strain selection:
Rat is one of the recommended species by regulatory agencies for conducting preclinical toxicological studies among rodents
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: In-house
- Females nulliparous and non-pregnant: yes
- Age at study initiation: 7 weeks
- Weight at study initiation: Males: 130.07 g to 149.99 g
Females: 122.05 g to 140.00 g
- Housing: L430 X B285 X H150 mm
- Diet (e.g. ad libitum): Altromin maintenance diet for rats and mice (manufactured by Altromin Spezialfutter GmbH & Co. KG)
- Water (e.g. ad libitum): Deep bore-well water passed through Reverse Osmosis Unit
- Acclimation period: 06 April 2018 to 11 April 2018


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.4°C to 23.3°C
- Humidity (%): 41% to 66%
- 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: 12 april 2018 To: 09 May 2019
Route of administration:
oral: gavage
Details on route of administration:
The test item was administered through oral route using stainless steel (gavage) cannula as it is the probable route in human
Vehicle:
corn oil
Details on oral exposure:

VEHICLE
- Justification for use and choice of vehicle (if other than water): Corn oil was used as vehicle for the preparation of formulations as evidenced by the in-house miscibility test results. Corn oil is the universally accepted vehicle in preclinical formulations

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 done by Analytical Chemistry department of Bioneeds India Private Limited. The analysis was done as per methods detailed in the Study Plan No. BIO-ANM 813 and the results are presented in the report. Sampling and analysis of formulations was performed during week 1 and week 4 of the treatment. The samples were collected in duplicates from top, middle and bottom layer from low, mid and high dose concentrations, in duplicates from middle layer from vehicle control.

The collected samples were transferred to Analytical Chemistry department of Bioneeds India Private Limited for dose concentration analysis. One set of aliquot of each formulation was analyzed. The second set of samples was discarded as the analysis results of first set of samples were within the range of 85 to 115% of the nominal concentration and the relative standard deviation (% RSD) was ≤10%.
Duration of treatment / exposure:
28 consecutive days
Frequency of treatment:
28 consecutive days
Dose / conc.:
0 mg/kg bw/day (nominal)
Remarks:
Vehicle control / Vehicle control recovery
Dose / conc.:
100 mg/kg bw/day (nominal)
Remarks:
Low dose
Dose / conc.:
300 mg/kg bw/day (nominal)
Remarks:
Mid dose
Dose / conc.:
1 000 mg/kg bw/day (nominal)
Remarks:
High dose/ High dose recovery
No. of animals per sex per dose:
5 males and 5 females per sex per group
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: The doses of 100, 300 and 1000 mg/kg body weight was selected as low (G2), mid (G3) and high dose (G4/G4R) levels for test item Hostacor ITD based on the results obtained from the dose range finding study (Bioneeds Study Number: BIO-TX 3133).

- Rationale for animal assignment: randomised

- Post-exposure recovery period in satellite groups: 14 days recovery period
Positive control:
Not applicable
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Once daily
- Cage side observations checked in table [No.1] were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly once

BODY WEIGHT: Yes
- Time schedule for examinations: weekly once

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: Yes

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: before start of the treatment and during week 4 of treatment for G1 and G4 group animals and during week 6 for recovery group animals (G1R and G4R)
- Dose groups that were examined: G1/G1R and G4/G4R

HAEMATOLOGY: Yes
- Time schedule for collection of blood: main groups on day 29 and from recovery group animals on day 43
- Anaesthetic used for blood collection: Yes
- Animals fasted: Yes
- How many animals: all the animals
- Parameters checked in table [No.11] were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: main groups on day 29 and from recovery group animals on day 43
- Animals fasted: Yes
- How many animals: all the animals
- Parameters checked in table [No.12] were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: main groups on day 29 and from recovery group animals on day 43
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters checked in table [No.13] were examined.

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: during week 4 for all the main group animals and during week 6 for recovery group animals (G1R and G4R)
- Dose groups that were examined: G1 to G4, G1R and G4R
- Battery of functions tested: sensory activity / grip strength / motor activity

IMMUNOLOGY: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes (see table 16)

HISTOPATHOLOGY: Yes (see table 17)
Statistics:
SPSS software version 22. One way ANOVA followed by Dunnett’s post test were done for different treatment and recovery groups comparing with the respective control group data. Data of recovery groups during recovery period was subjected to student’s t-test.
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
No relevant clinical signs of toxicity was observed in any of the vehicle or test item administered animals up to the high dose group. A slight wet perineum was observed in a few high dose test item administered animals (males: 2/5 and females: 2/5 in main group animals; males: 1/5 and females: 2/5 in recovery group animals) which is not considered to be of toxicological relevance
Mortality:
mortality observed, non-treatment-related
Description (incidence):
No relevant clinical signs of toxicity was observed in any of the vehicle or test item administered animals up to the high dose group. A slight wet perineum was observed in a few high dose test item administered animals (males: 2/5 and females: 2/5 in main group animals; males: 1/5 and females: 2/5 in recovery group animals) which is not considered to be of toxicological relevance
Body weight and weight changes:
effects observed, non-treatment-related
Description (incidence and severity):
No treatment related changes in mean body weight and body weight gain (percent change in body weight with respect to Day 1) were noted. However, statistical significant decrease in mean body weight was observed on days 8, 15, 22 and 28 in G4 males and statistical significant decrease in percent change in body weight with respect to Day 1 was observed in G4 males during Day 1-8 to Day 1-28, and in G4R females during Day 1-8. All these changes are considered as incidental due to lack of dose dependency and also no concomitant changes were observed in feed consumption
Food efficiency:
no effects observed
Ophthalmological findings:
no effects observed
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
No treatment related changes in hematology parameters were noted.

A non-dose dependent statistically significant increase in percent and absolute monocytes in G3 males; increase in percent and absolute neutrophils in G4 males; decrease in percent lymphocytes in G4 males was observed. Statistical significant increase in percent and absolute neutrophils in G4 females; increase in percent and absolute monocytes in G4 females; decrease in percent lymphocytes in G4 females was observed. However, in the absence of significant variations in total leukocyte counts, the observed variations are considered incidental and also no such changes were noted at the end of recovery period.

At the end of recovery period, statistical significant increase in Haemoglobin, Mean corpuscular hemoglobin and Mean corpuscular haemoglobin concentration observed in G4 females are considered incidental and not related to treatment as these range of values are minimal and also similar changes were not noticed in main group animals
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
In males, the non-dose dependent decrease in creatinine and A/G ratio in G3; decrease in urea and BUN in G2 and G3; decrease in cholesterol and calcium in G4; increase in ALP in G3 and increase in AST in G4 was observed. In females, statistically significant decrease in cholesterol, total protein, cholinesterase and globulin in G4; statistically significant increase ALP in G2, G3 and G4; increase in total bilirubin and ALT in G4 was noted. All these clinical chemistry variations are considered as incidental as there were no corresponding histopathological findings observed at high dose group animals
Urinalysis findings:
no effects observed
Behaviour (functional findings):
effects observed, non-treatment-related
Description (incidence and severity):
No treatment related changes were observed in neurological/functional examination battery carried out during Week 4 for main group and Week 6 for recovery group animals. Statistical significant decrease in movement counts noted in G4 males was considered incidental due to lack of dose dependency and also none of the neuromuscular parameters
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
In males, statistically significant increase in absolute and relative spleen weight in G2, increase in absolute and relative weight of Prostate+Seminal vesicles and coagulating gland in G3, increase in absolute epididymis and testes weight in G3, increase in absolute heart, kidneys and liver weight in G2 and G3 was noted. The statistical significant decrease in fasting body weight in G4, increase in relative kidneys and liver weight in G2, G3 and G4, increase in relative heart weight in G3 and increase in relative epididymis and testes weight in G4 was noted.

In females, statistical significant increase in absolute and relative kidneys weight in G4, increase in absolute liver weight in G4 and relative liver weight in G3 and G4 was noted.

All these observed changes are considered as incidental and not related to treatment as there were no corresponding histopathological findings observed at the high dose group. Also, a statistical significant increase in absolute and relative adrenals weight was observed in G4R animals is considered as incidental not related to treatment as these effects were not observed in main group animals
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Lesions observed in animals from high dose group were MNC infiltration in liver, tubular basophilia in kidneys and perivascular MNC infiltration in lungs. Similar findings were also noticed in concurrent control group. Hence, these lesions were considered as spontaneous or background lesions.
A detailed qualitative examination of the testes was made taking into account the tubular stages of the spermatogenic cycle. The examination was conducted in order to identify treatment related effects such as missing germ cell layers or types, retained spermatids, multinucleate or apoptotic germ cells and sloughing of spermatogenic cells into the lumen or any cell or stage specificity of testicular findings
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Remarks on result:
not determinable due to absence of adverse toxic effects
Key result
Critical effects observed:
no

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

 

Group, Sex & Dose

(mg/kg body weight/day)

No. of

Animals

Clinical Sign(s)/

Number of Animals revealed

Mortality                               (No. of incidence/

No. of animals)

G1, M & 0

5

N

0/5

G2, M & 100

5

N

0/5

G3, M & 300

5

N

0/5

G4, M & 1000

5

73+/2, N/3

0/5

G1R, M & 0

5

N

0/5

G4R, M & 1000

5

73+/1, N/4

0/5

G: Group; M: Male; N: Normal; 73: Wet perineum; +: Slight; R: Recovery.

 

                  


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

 

Group, Sex & Dose

(mg/kg body weight/day)

No. of

Animals

Clinical Sign(s)/

Number of Animals revealed

Mortality                               (No. of incidence/

No. of animals)

G1, F & 0

5

N

0/5

G2, F & 100

5

N

0/5

G3, F & 300

5

N

0/5

G4, F & 1000

5

73+/2, N/3

0/5

G1R, F & 0

5

N

0/5

G4R, F & 1000

5

73+/2, N/3

0/5

G: Group; F: Female; N: Normal; 73: Wet perineum; +: Slight; R: Recovery.


 

TABLE 2.  SUMMARY OF DETAILED CLINICAL EXAMINATION RECORD

Group: G4                                         Dose: 1000 mg/kg body weight/day                Days: 15, 22 & 28

                                                                                                                                                                    

Examination         

Sex

Male & Female

1. Skin & Fur

0

2. Eye Discharge

0

3. Mucous Membrane

0

4. Occurrence of Secretions & Excretions

a. Salivation

0

b. Urine Staining

1* (2-M & 2-F)

c. Fecal Staining or Diarrhoea

0

d. Nasal Discharge

0

5. Autonomic Activity

a. Lacrimation

0

b. Piloerection

0

c. Pupillary Response

1

d. Un-usual Respiratory Pattern

0

6. Response to Handling

1

7. Changes in Gait

0

8. Posture

0

9. Clonic or Tonic Movements

0

10. Stereotypies

a. Repetitive Circling

0

b. Excessive Grooming

0

11.Bizarre behavior

a. Self-mutilation

0

b. Walking Backwards

0

12. Hair Thinning with Hair Re-growth

0

 0: Absent; 1: Present; *: slight wet perineum; M: Male; F: Female.


 

TABLE 2 (Contd..,)  SUMMARY OF DETAILED CLINICAL EXAMINATION RECORD

Group: G4R                                         Dose: 1000 mg/kg body weight/day              Days: 15, 22 & 28

 

Examination         

Sex

Male & Female

1. Skin & Fur

0

2. Eye Discharge

0

3. Mucous Membrane

0

4. Occurrence of Secretions & Excretions

a. Salivation

0

b. Urine Staining

1* (1-M & 2-F)

c. Fecal Staining or Diarrhoea

0

d. Nasal Discharge

0

5. Autonomic Activity

a. Lacrimation

0

b. Piloerection

0

c. Pupillary Response

1

d. Un-usual Respiratory Pattern

0

6. Response to Handling

1

7. Changes in Gait

0

8. Posture

0

9. Clonic or Tonic Movements

0

10. Stereotypies

a. Repetitive Circling

0

b. Excessive Grooming

0

11.Bizarre behavior

a. Self-mutilation

0

b. Walking Backwards

0

12. Hair Thinning with Hair Re-growth

0

 0: Absent; 1: Present; *: slight wet perineum; M: Male; F: Female.

 


TABLE 3.SUMMARY OF BODY WEIGHTS (g)RECORD

 

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

Body Weight (g) on Days

1

8

15

22

28

35

42

G1, M & 0

Mean

174.07

209.29

236.88

271.53

291.43

-

-

±SD

8.29

12.08

18.65

22.28

27.55

-

-

n

5

5

5

5

5

-

-

G2, M & 100

Mean

172.73

211.66

239.60

266.86

293.48

-

-

±SD

7.91

12.32

18.56

19.06

21.78

-

-

n

5

5

5

5

5

-

-

G3, M & 300

Mean

176.21

212.58

244.39

273.80

299.72

-

-

±SD

7.30

12.37

15.26

14.44

17.50

-

-

n

5

5

5

5

5

-

-

G4, M & 1000

Mean

176.36

189.75*

206.52*

224.04*

251.00*

-

-

±SD

8.17

5.69

13.62

12.48

12.31

-

-

n

5

5

5

5

5

-

-

G1R, M & 0

Mean

174.11

208.03

243.25

267.42

295.11

336.49

344.25

±SD

8.39

15.14

22.49

26.45

26.35

32.11

33.85

n

5

5

5

5

5

5

5

G4R, M & 1000

Mean

173.58

200.74

225.96

257.62

279.12

320.29

326.71

±SD

6.03

9.39

14.13

16.03

20.04

18.47

19.00

n

5

5

5

5

5

5

5

G: Group; M: Male; R: Recovery; SD: Standard Deviation; n: Number of animals; -: Not Applicable

*: Statistically significant (p<0.05)

 

 

 

 

 

 

 

 

 

 


 

TABLE 3 (Contd..,)  SUMMARY OF BODY WEIGHTS (g) RECORD

 

 

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

Body Weight (g) on Days

1

8

15

22

28

35

42

G1, F & 0

Mean

154.53

176.53

195.19

208.48

221.06

-

-

±SD

8.20

11.12

17.10

16.78

18.79

-

-

n

5

5

5

5

5

-

-

G2, F & 100

Mean

156.49

181.40

197.76

209.43

220.60

-

-

±SD

6.28

5.96

6.66

6.75

8.44

-

-

n

5

5

5

5

5

-

-

G3, F & 300

Mean

156.49

180.41

195.66

208.53

223.71

-

-

±SD

9.18

9.88

6.24

7.03

6.92

-

-

n

5

5

5

5

5

-

-

G4, F & 1000

Mean

156.21

171.64

187.95

202.06

215.70

-

-

±SD

8.41

11.37

17.06

16.75

16.90

-

-

n

5

5

5

5

5

-

-

G1R, F & 0

Mean

155.81

179.12

191.93

207.70

218.68

245.02

248.74

±SD

5.45

9.37

9.72

9.29

10.75

9.75

10.14

n

5

5

5

5

5

5

5

G4R, F & 1000

Mean

156.07

169.48

184.50

201.39

219.60

246.26

249.13

±SD

6.50

8.50

6.25

12.48

18.03

15.06

15.46

n

5

5

5

5

5

5

5

G: Group; F: Female; R: Recovery; SD: Standard Deviation; n: Number of animals; -: Not Applicable

 

 

 

TABLE 4.  SUMMARY OF PERCENT CHANGE IN BODY WEIGHT (%) WITH RESPECT TO DAY 1RECORD

 

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

Percent Change in Body Weight (%) during Days

1 to 8

1 to 15

1 to 22

1 to 28

1 to 35

1 to 42

G1, M & 0

Mean

20.19

35.96

55.83

67.20

-

-

±SD

1.64

5.42

6.65

9.42

-

-

n

5

5

5

5

-

-

G2, M & 100

Mean

22.50

38.58

54.39

69.79

-

-

±SD

2.96

5.41

5.54

6.96

-

-

n

5

5

5

5

-

-

G3, M & 300

Mean

20.61

38.66

55.37

70.02

-

-

±SD

3.66

5.24

4.18

4.10

-

-

n

5

5

5

5

-

-

G4, M & 1000

Mean

7.68*

17.31*

27.22*

42.56*

-

-

±SD

2.96

9.75

8.82

9.65

-

-

n

5

5

5

5

-

-

G1R, M & 0

Mean

19.39

39.56

53.39

69.30

93.05

97.47

±SD

4.00

8.03

9.64

8.73

12.03

12.73

n

5

5

5

5

5

5

G4R, M & 1000

Mean

15.64

30.09

48.33

60.68

84.47

88.15

±SD

3.01

4.44

5.06

7.28

7.01

6.57

n

5

5

5

5

5

5

 G: Group; M: Male; R: Recovery; SD: Standard Deviation; n: Number of animals; -: Not Applicable

 *: Statistically significant (p<0.05)

 

 


TABLE 4 (Contd..,).  SUMMARY OF PERCENT CHANGE IN BODY WEIGHT (%) WITH RESPECT TO DAY 1 RECORD  

 

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

Percent Change in Body Weight (%) during Days

1 to 8

1 to 15

1 to 22

1 to 28

1 to 35

1 to 42

G1, F & 0

Mean

14.21

26.24

34.85

42.98

-

-

±SD

2.40

7.10

6.58

7.67

-

-

n

5

5

5

5

-

-

G2, F & 100

Mean

15.96

26.44

33.91

41.04

-

-

±SD

2.47

3.63

3.98

4.66

-

-

n

5

5

5

5

-

-

G3, F & 300

Mean

15.36

25.21

33.44

43.24

-

-

±SD

3.79

4.34

4.57

7.27

-

-

n

5

5

5

5

-

-

G4, F & 1000

Mean

9.84

20.25

29.33

38.09

-

-

±SD

2.31

7.44

7.44

7.89

-

-

n

5

5

5

5

-

-

G1R, F & 0

Mean

14.91

23.14

33.36

40.39

57.35

59.72

±SD

2.94

2.97

5.66

6.36

6.93

6.46

n

5

5

5

5

5

5

G4R, F & 1000

Mean

8.61*

18.30

29.03

40.60

57.77

59.64

±SD

3.58

3.73

5.36

7.90

6.43

7.30

n

5

5

5

5

5

5

 G: Group; F: Female; R: Recovery; SD: Standard Deviation; n: Number of animals; -: Not Applicable

 *: Statistically significant (p<0.05)

 

 

 

 


TABLE 5.  SUMMARY OF FEED CONSUMPTION (g/rat/day)RECORD

 

Group, Sex & Dose

(mg/kg body weight/day)

Week 1

Week 2

Week 3

Week 4

Week 5

Week 6

G1, M & 0

Mean

19.22

19.40

20.06

22.48

-

-

±SD

3.62

3.27

3.96

3.49

-

-

n

2

2

2

2

-

-

G2, M & 100

Mean

17.79

17.97

18.30

21.21

-

-

±SD

2.67

2.62

2.97

2.61

-

-

n

2

2

2

2

-

-

G3, M & 300

Mean

17.98

18.27

18.95

21.04

-

-

±SD

2.80

2.29

2.54

2.10

-

-

n

2

2

2

2

-

-

G4, M & 1000

Mean

16.03

16.10

17.28

20.18

-

-

±SD

1.04

0.51

0.73

1.03

-

-

n

2

2

2

2

-

-

G1R, M & 0

Mean

19.42

19.60

20.31

22.32

20.23

19.44

±SD

3.73

4.15

4.52

4.66

3.82

3.27

n

2

2

2

2

2

2

G4R, M & 1000

Mean

16.31

16.86

17.57

20.50

19.14

18.20

±SD

0.74

0.90

1.78

2.61

3.39

2.99

n

2

2

2

2

2

2

 G: Group; M: Male; R: Recovery; SD: Standard Deviation, n: Number of cages; -: Not Applicable

 

 

 


TABLE 5 (Contd..,).  SUMMARY OF FEED CONSUMPTION (g/rat/day)RECORD

 

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

Week 1

Week 2

Week 3

Week 4

Week 5

Week 6

G1, F & 0

Mean

16.14

17.17

17.65

19.61

-

-

±SD

1.22

1.33

1.36

1.74

-

-

n

2

2

2

2

-

-

G2, F & 100

Mean

14.55

14.55

14.98

16.62

-

-

±SD

1.75

1.55

1.99

1.84

-

-

n

2

2

2

2

-

-

G3, F & 300

Mean

13.74

14.59

15.37

17.52

-

-

±SD

1.47

1.24

1.58

2.24

-

-

n

2

2

2

2

-

-

G4, F & 1000

Mean

12.05

13.05

12.98

14.48

-

-

±SD

1.06

2.17

2.21

1.83

-

-

n

2

2

2

2

-

-

G1R, F & 0

Mean

14.72

15.57

16.46

17.22

16.48

16.57

±SD

0.92

1.09

1.08

1.57

1.74

0.66

n

2

2

2

2

2

2

G4R, F & 1000

Mean

11.88

12.55

12.31

14.67

13.58

14.04

±SD

0.76

1.47

1.30

2.55

2.16

2.52

n

2

2

2

2

2

2

 G: Group; F: Female; R: Recovery; SD: Standard Deviation; n: Number of cages; -: Not Applicable

 

 

 


TABLE 6.  SUMMARY OF OPHTHALMOSCOPIC EXAMINATION RECORD

 

Week 4

Group & Sex

G1 & M

G4 & M

G1 & F

G4 & F

Dose (mg/kg body weight/day)

0

1000

0

1000

No. of Animals

5

5

5

5

Observations

Eye Parameters ↓

L

R

L

R

L

R

L

R

Adnexa

Lids

N

N

N

N

N

N

N

N

Ducts

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

Vessels

N

N

N

N

N

N

N

N

Macula

N

N

N

N

N

N

N

N

Optic disc

N

N

N

N

N

N

N

N

Tapetum lucidum

N

N

N

N

N

N

N

N

Tapetum nigrum

N

N

N

N

N

N

N

N

Other findings

N

N

N

N

N

N

N

N

G: Group; M: Male; F: Female; N: No abnormality detected; L: Left; R: Right.


 

TABLE 6 (Contd..,). SUMMARY OF OPHTHALMOSCOPIC EXAMINATION RECORD

 

Week 6

Group & Sex

G1R & M

G4R & M

G1R & F

G4R & F

Dose (mg/kg body weight/day)

0

1000

0

1000

No. of Animals

5

5

5

5

Observations

Eye Parameters ↓

L

R

L

R

L

R

L

R

Adnexa

Lids

N

N

N

N

N

N

N

N

Ducts

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

Vessels

N

N

N

N

N

N

N

N

Macula

N

N

N

N

N

N

N

N

Optic disc

N

N

N

N

N

N

N

N

Tapetum lucidum

N

N

N

N

N

N

N

N

Tapetum nigrum

N

N

N

N

N

N

N

N

Other findings

N

N

N

N

N

N

N

N

G: Group; M: Male; F: Female; N: No abnormality detected; L: Left; R: Right

 

 

 

TABLE 7.  SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB)RECORD

 

Parameters↓

 

Week 4

Week 6

Group & Sex

G1 & M

G2 & M

G3 & M

G4 & M

G1R & M

G4R & M

Dose (mg/kg body weight/day)

0

100

300

1000

0

1000

Number of Animals

5

5

5

5

5

5

Home cage observations

a. Convulsions

1

1

1

1

1

1

b. Tremors

1

1

1

1

1

1

c. Palpebral closure

1

1

1

1

1

1

Handling observation

a. Ease of removal from the cage

2

2

2

2

2

2

b. Ease of handling animal in hand

2

2

2

2

2

2

c. Lacrimation

1

1

1

1

1

1

d. Red deposits around

 

Eyes

1

1

1

1

1

1

Nose

1

1

1

1

1

1

Mouth

1

1

1

1

1

1

e. Crusty

depositsaround

 

Eyes

1

1

1

1

1

1

Nose

1

1

1

1

1

1

Mouth

1

1

1

1

1

1

f. Salivation

1

1

1

1

1

1

g. Fur appearance

1

1

1

1

1

1

h. Piloerection

1

1

1

1

1

1

i. Palpebral Closure

1

1

1

1

1

1

j. RespiratoryCharacter

1

1

1

1

1

1

k. EyeProminence

1

1

1

1

1

1

l. Muscle Tone

1

1

1

1

1

1

Home cage observations: a. Convulsions -1=Absentb. Tremors-1=Absentc. Palpebral closure -1=Normal, Handling observation : a. Ease of removal from the cage -2=Normal, b. Ease of handling animal in hand -2=Normal, c. Lacrimation -1=None, d. Red deposits around -1=Absente. Crusty deposits around -1=Absentf. Salivation-1=Normal, g. Fur Appearance -1=Normal, h. Piloerection -1=None, i. Palpebral Closure -1=Normal, j. Respiratory character -1=Normal, k. Eye Prominence -1=Normal, l. Muscle tone -1=Normal

G: Group; M: Male; R: Recovery;


TABLE 7 (Contd..,). SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB) RECORD

 

 

Parameters↓

 

Week 4

Week 6

Group & Sex

G1 & M

G2 & M

G3 & M

G4 & M

G1R & M

G4R & M

Dose (mg/kg body weight/day)

0

100

300

1000

0

1000

Number of Animals

5

5

5

5

5

5

Open field Observation

a. Mobility

1

1

1

1

1

1

b. Gait

1

1

1

1

1

1

c. Arousal

3

3

3

3

3

3

d. Rearing

Mean

4.2

3.8

3.6

4.2

4.0

4.0

±SD

0.8

0.8

0.9

0.8

0.7

0.7

e. Urination

Numbers of pools of urine

Mean

2.4

2.4

2.0

2.4

1.6

1.4

±SD

1.1

1.1

0.7

1.1

0.5

0.9

f. Defecation

Number of fecal pellets

Mean

1.8

1.8

1.6

2.0

1.2

1.2

±SD

0.4

0.4

0.5

0.7

0.8

0.8

g. Stereotypies- repetitive circling

1

1

1

1

1

1

h. Excessive grooming

Mean

2.6

2.4

2.4

2.6

3.2

3.4

±SD

0.5

0.5

0.5

0.5

0.8

0.5

Sensory observations

a.Startle response

2

2

2

2

2

2

b. Touch response

2

2

2

2

2

2

c. Pupil response

2

2

2

2

2

2

d. Response to nociceptive stimuli

2

2

2

2

2

2

e. Righting reflex

1

1

1

1

1

1

Physiological observation

Body temperature (°F)

Mean

98.8

99.3

99.0

99.0

99.4

99.3

±SD

0.6

0.4

0.6

0.6

0.7

0.4

Open field Observation: a. Mobility- 1=Normal,b. Gait- 1=Normal,c. Arousal- 3=Normal, g. Stereotypies- repetitive circling - 1=Absent,Sensory Observations: a. Startle Response-2=Normal,b. Touch Response- 2=Normal,c. Pupil Response2=Normal,d. Response to Nociceptive stimuli- 2=Normal, e. Righting Reflex- 1=Present

    G: Group; M: Male; R: Recovery; Slight were perineum was noted in G4M (2/5) and G4F (2/5)


 

TABLE 7 (Contd..,). SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB) RECORD

 

 

Parameters↓

 

Week 4

Week 6

 

Group & Sex

G1 & F

G2 & F

G3 & F

G4 & F

G1R & F

G4R & F

 

Dose (mg/kg body weight/day)

0

100

300

1000

0

1000

 

Number of Animals

5

5

5

5

5

5

 

Home cage observations

a. Convulsions

1

1

1

1

1

1

 

b. Tremors

1

1

1

1

1

1

 

c. Palpebral closure

1

1

1

1

1

1

 

Handling observation

a. Ease of removal from the cage

2

2

2

2

2

2

 

b. Ease of handling animal in hand

2

2

2

2

2

2

 

c. Lacrimation

1

1

1

1

1

1

 

d. Red deposits around

 

Eyes

1

1

1

1

1

1

 

Nose

1

1

1

1

1

1

 

Mouth

1

1

1

1

1

1

 

e. Crusty

depositsaround

 

Eyes

1

1

1

1

1

1

 

Nose

1

1

1

1

1

1

 

Mouth

1

1

1

1

1

1

 

f. Salivation

1

1

1

1

1

1

 

g. Fur appearance

1

1

1

1

1

1

 

h. Piloerection

1

1

1

1

1

1

 

i. Palpebral Closure

1

1

1

1

1

1

 

j. RespiratoryCharacter

1

1

1

1

1

1

 

k. EyeProminence

1

1

1

1

1

1

 

l. Muscle Tone

1

1

1

1

1

1

 

Home cage observations: a. Convulsions -1=Absentb. Tremors-1=Absentc. Palpebral closure -1=Normal, Handling observation : a. Ease of removal from the cage -2=Normal, b. Ease of handling animal in hand -2=Normal, c. Lacrimation -1=None, d. Red deposits around -1=Absente. Crusty deposits around -1=Absentf. Salivation-1=Normal, g. Fur Appearance -1=Normal, h. Piloerection -1=None, i. Palpebral Closure -1=Normal, j. Respiratory character -1=Normal, k. Eye Prominence -1=Normal, l. Muscle tone -1=Normal

G: Group; F: Female; R: Recovery


 

TABLE 7 (Contd..,). SUMMARY OF NEUROLOGICAL/FUNCTIONAL OBSERVATION BATTERY (FOB) RECORD

 

 

Parameters↓

 

Week 4

Week 6

 

Group & Sex

G1 & F

G2 & F

G3 & F

G4 & F

G1R & F

G4R & F

 

Dose (mg/kg body weight/day)

0

100

300

1000

0

1000

 

Number of Animals

5

5

5

5

5

5

 

Open field Observation

a. Mobility

1

1

1

1

1

1

 

b. Gait

1

1

1

1

1

1

 

c. Arousal

3

3

3

3

3

3

 

d. Rearing

Mean

3.8

3.6

3.4

3.8

4.0

4.0

 

±SD

0.8

0.5

0.5

0.8

0.7

0.7

 

e. Urination

Numbers of pools of urine

Mean

1.6

2.0

1.8

1.6

1.4

1.4

 

±SD

1.1

0.7

0.8

1.1

0.5

0.5

 

f. Defecation

Number of fecal pellets

Mean

1.4

1.6

1.6

1.4

1.4

1.4

 

±SD

0.9

0.5

0.5

0.9

0.5

0.5

 

g. Stereotypies- repetitive circling

1

1

1

1

1

1

 

h. Excessive grooming

Mean

2.6

2.8

2.6

2.4

2.8

3.0

 

±SD

0.5

0.4

0.5

0.5

0.4

0.7

 

Sensory observations

a.Startle response

2

2

2

2

2

2

 

b. Touch response

2

2

2

2

2

2

 

c. Pupil response

2

2

2

2

2

2

 

d. Response to nociceptive stimuli

2

2

2

2

2

2

 

e. Righting reflex

1

1

1

1

1

1

 

Physiological observation

Body temperature (°F)

Mean

99.6

99.2

99.2

99.7

99.5

99.3

 

±SD

0.7

0.9

0.8

0.5

0.8

0.7

 

Open field Observation: a. Mobility- 1=Normal,b. Gait- 1=Normal,c. Arousal- 3=Normal, g. Stereotypies- repetitive circling - 1=Absent,Sensory Observations: a. Startle Response-2=Normal,b. Touch Response- 2=Normal,c. Pupil Response2=Normal,d. Response to Nociceptive stimuli- 2=Normal, e. Righting Reflex- 1=Present

    G: Group; F: Female; R: Recovery

 


 


TABLE 8.  SUMMARY OF LANDING FOOT SPLAY (cm) RECORD

 

Group, Sex & Dose

(mg/kg body weight/day)

Hind Limb Foot Splay (cm)

G1, M & 0

Mean

6.1

±SD

0.4

n

5

G2, M & 100

Mean

6.1

±SD

0.3

n

5

G3, M & 300

Mean

6.4

±SD

0.3

n

5

G4, M & 1000

Mean

6.4

±SD

0.3

n

5

G1, F & 0

Mean

5.5

±SD

0.9

n

5

G2, F & 100

Mean

5.4

±SD

0.3

n

5

G3, F & 300

Mean

5.2

±SD

0.2

n

5

G4, F & 1000

Mean

5.2

±SD

0.3

n

5

G1R, M & 0

Mean

6.6

±SD

0.8

n

5

G4R, M & 1000

Mean

6.5

±SD

0.5

n

5

G1R, F & 0

Mean

5.3

±SD

0.8

n

5

G4R, F & 1000

Mean

5.4

±SD

0.5

n

5

G: Group; M: Male; F: Female; R: Recovery; SD: Standard Deviation; n: number of animals

 


TABLE 9.  SUMMARY OF PHOTOACTOMETER READING RECORD

 

Group, Sex & Dose

(mg/kg body weight/day)

Movement counts

G1, M & 0

Mean

619.2

±SD

41.6

n

5

G2, M & 100

Mean

603.6

±SD

22.9

n

5

G3, M & 300

Mean

561.6

±SD

91.1

n

5

G4, M & 1000

Mean

523.0*

±SD

52.2

n

5

G1, F & 0

Mean

665.6

±SD

35.4

n

5

G2, F & 100

Mean

600.4

±SD

45.7

n

5

G3, F & 300

Mean

625.6

±SD

55.6

n

5

G4, F & 1000

Mean

584.6

±SD

70.9

n

5

G1R, M & 0

Mean

653.4

±SD

50.4

n

5

G4R, M & 1000

Mean

580.8

±SD

66.9

n

5

G1R, F & 0

Mean

711.4

±SD

55.0

n

5

G4R, F & 1000

Mean

697.8

±SD

43.1

n

5

G: Group; M: Male; F: Female; R: Recovery; SD: Standard Deviation; n: number of animals;

*: Statistically significant (p<0.05)

 

 

TABLE 10.SUMMARY OF GRIP STRENGTH (kgf) MEASUREMENT RECORD

 

Group, Sex & Dose

(mg/kg body weight/day)

Fore limb

Hind limb

G1, M & 0

Mean

1.511

0.526

±SD

0.041

0.027

n

5

5

G2, M & 100

Mean

1.525

0.518

±SD

0.040

0.025

n

5

5

G3, M & 300

Mean

1.523

0.529

±SD

0.040

0.020

n

5

5

G4, M & 1000

Mean

1.507

0.516

±SD

0.032

0.037

n

5

5

G1, F & 0

Mean

1.518

0.510

±SD

0.023

0.040

n

5

5

G2, F & 100

Mean

1.531

0.536

±SD

0.024

0.025

n

5

5

G3, F & 300

Mean

1.541

0.550

±SD

0.024

0.020

n

5

5

G4, F & 1000

Mean

1.516

0.502

±SD

0.015

0.039

n

5

5

G1R, M & 0

Mean

1.493

0.506

±SD

0.016

0.053

n

5

5

G4R, M & 1000

Mean

1.487

0.505

±SD

0.019

0.043

n

5

5

G1R, F & 0

Mean

1.483

0.489

±SD

0.034

0.042

n

5

5

G4R, F & 1000

Mean

1.478

0.481

±SD

0.026

0.053

n

5

5

 G: Group; M: Male; F: Female; R: Recovery; SD: Standard Deviation; n: number of animals.

 


 


TABLE 11.   SUMMARY OF HAEMATOLOGY RECORD

 

                Group, Sex & Dose

(mg/kg body weight/day)

Total Leucocyte Count

Total Erythrocyte Count

Hemoglobin

Haematocrit

Mean Corpuscular Volume

Mean Corpuscular Hemoglobin

Mean Corpuscular Hemoglobin 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

8.88

7.15

13.72

42.30

59.22

19.20

32.40

917.60

7.44

±SD

3.00

0.38

0.54

1.48

1.70

0.56

0.24

215.31

0.53

n

5

5

5

5

5

5

5

5

5

G2, M & 100

Mean

8.85

7.41

13.96

43.18

58.32

18.88

32.38

890.40

7.54

±SD

3.67

0.40

0.68

1.98

1.52

0.39

0.45

66.20

0.17

n

5

5

5

5

5

5

5

5

5

G3, M & 300

Mean

10.74

7.35

13.18

41.56

56.86

18.08

31.68

916.00

8.58

±SD

1.72

0.48

1.76

3.05

6.98

3.05

2.16

290.00

1.60

n

5

5

5

5

5

5

5

5

5

G4, M & 1000

Mean

10.86

6.94

12.36

40.60

58.58

17.72

30.30

882.20

8.02

±SD

2.93

0.33

2.43

1.50

1.12

3.22

5.56

101.56

0.55

n

5

5

5

5

5

5

5

5

5

G1R, M & 0

Mean

10.64

7.10

13.42

40.18

56.74

18.98

33.42

983.20

7.56

±SD

2.48

0.59

0.73

2.12

2.13

1.30

1.22

110.08

0.21

n

5

5

5

5

5

5

5

5

5

G4R, M & 1000

Mean

9.93

6.85

13.28

39.48

57.66

19.40

33.60

1007.40

7.64

±SD

1.56

0.15

0.26

0.73

0.29

0.30

0.51

68.20

0.38

n

5

5

5

5

5

5

5

5

5

                  G: Group; M: Male; R: Recovery; SD: Standard Deviation; n:number of animals.

 

 

TABLE 11 (Contd..,).  SUMMARY OF HAEMATOLOGY RECORD

 

Group, Sex & Dose

(mg/kg body weight/day)

Reticulocyte Count

Neutrophils

Lymphocytes

Monocytes

Eosinophils

Basophils

Absolute Reticulocyte Count

(Retic)

(Neut)

(Lymph)

(Mono)

(Eos)

(Baso)

(Retic)

(%)

(%)

(%)

(%)

(%)

(%)

(109cells/L)

G1, M & 0

Mean

1.86

21.46

73.04

3.16

0.78

0.58

133.36

±SD

0.52

4.08

4.68

0.58

0.30

0.13

38.89

n

5

5

5

5

5

5

5

G2, M & 100

Mean

1.66

22.36

72.26

3.22

0.56

0.54

122.72

±SD

0.30

4.44

4.17

0.27

0.15

0.11

21.50

n

5

5

5

5

5

5

5

G3, M & 300

Mean

2.11

19.40

72.80

4.78*

1.16

0.48

156.70

±SD

0.66

2.28

3.53

1.29

0.45

0.15

56.89

n

5

5

5

5

5

5

5

G4, M & 1000

Mean

1.43

35.58*

57.80*

4.00

0.52

0.48

98.70

±SD

0.39

10.83

9.60

0.92

0.28

0.25

23.42

n

5

5

5

5

5

5

5

G1R, M & 0

Mean

2.04

23.58

72.16

2.06

0.62

0.48

145.42

±SD

0.27

7.88

8.27

0.63

0.08

0.16

29.01

n

5

5

5

5

5

5

5

G4R, M & 1000

Mean

1.97

17.08

78.26

2.64

0.66

0.44

135.14

±SD

0.22

0.61

1.09

0.55

0.24

0.05

13.87

n

5

5

5

5

5

5

5

                   G: Group; M: Male; R: Recovery; SD: Standard Deviation; n:number of animals; *: Statistically significant (p<0.05)

TABLE 11 (Contd..,).  SUMMARY OF HAEMATOLOGY RECORD

 

Group, Sex & Dose

(mg/kg body weight/day)

Absolute Neutrophils

Absolute Lymphocytes

Absolute Monocytes

Absolute Eosinophils

Absolute Basophils

Prothrombin Time

Activated Prothrombin Time

(Neut)

(Lymph)

(Mono)

(Eos)

(Baso)

(PT)

(APTT)

(103cells/µL)

(103cells/µL)

(103cells/µL)

(103cells/µL)

(103cells/µL)

(Seconds)

(Seconds)

G1, M & 0

Mean

1.87

6.52

0.28

0.07

0.06

20.78

25.86

±SD

0.68

2.28

0.10

0.04

0.02

2.59

2.40

n

5

5

5

5

5

5

5

G2, M & 100

Mean

1.93

6.44

0.29

0.05

0.05

18.72

23.72

±SD

0.79

2.80

0.11

0.02

0.02

1.12

3.61

n

5

5

5

5

5

5

5

G3, M & 300

Mean

2.08

7.80

0.53*

0.12

0.06

17.58

25.48

±SD

0.39

1.23

0.21

0.05

0.02

2.95

3.46

n

5

5

5

5

5

5

5

G4, M & 1000

Mean

4.10*

6.07

0.43

0.05

0.05

18.66

23.20

±SD

2.47

0.77

0.12

0.02

0.02

1.90

3.39

n

5

5

5

5

5

5

5

G1R, M & 0

Mean

2.65

7.52

0.23

0.07

0.05

33.00

26.68

±SD

1.66

0.79

0.12

0.02

0.02

18.75

7.74

n

5

5

5

5

5

5

5

G4R, M & 1000

Mean

1.70

7.76

0.27

0.06

0.04

20.86

31.50

±SD

0.28

1.20

0.09

0.02

0.01

2.23

3.22

n

5

5

5

5

5

5

5

                   G: Group; M: Male; R: Recovery; SD: Standard Deviation; n:number of animals; *: Statistically significant (p<0.05)

 

 

                                       

TABLE 11 (Contd..,).  SUMMARY OF HAEMATOLOGY RECORD

 

Group, Sex & Dose

(mg/kg body weight/day)

Total Leucocyte Count

Total Erythrocyte Count

Hemoglobin

Haematocrit

Mean Corpuscular Volume

Mean Corpuscular Hemoglobin

Mean Corpuscular Hemoglobin 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

8.82

6.90

13.46

40.78

59.12

19.50

33.00

905.80

7.56

±SD

3.75

0.25

0.51

1.11

1.18

0.19

0.51

165.01

0.05

n

5

5

5

5

5

5

5

5

5

G2, F & 100

Mean

9.03

6.73

13.38

40.14

59.74

19.94

33.40

889.80

7.86

±SD

1.51

0.34

0.70

1.69

1.32

0.22

0.58

98.85

0.36

n

5

5

5

5

5

5

5

5

5

G3, F & 300

Mean

8.76

6.42

12.38

38.48

60.04

19.30

32.14

941.20

7.76

±SD

4.34

0.40

0.80

1.41

1.80

0.51

1.31

186.29

0.21

n

5

5

5

5

5

5

5

5

5

G4, F & 1000

Mean

12.08

7.06

13.50

41.38

58.70

19.14

32.60

1020.80

7.96

±SD

3.28

0.72

1.06

3.45

1.25

0.49

0.27

145.92

0.47

n

5

5

5

5

5

5

5

5

5

G1R, F & 0

Mean

8.76

6.81

13.10

38.36

56.42

19.28

34.14

914.80

7.32

±SD

3.15

0.30

0.28

1.11

1.24

0.83

0.70

138.18

0.19

n

5

5

5

5

5

5

5

5

5

G4R, F & 1000

Mean

9.69

6.74

13.80*

39.34

58.34

20.50*

35.18*

1033.20

7.20

±SD

1.64

0.18

0.34

1.05

2.03

0.72

0.18

83.29

0.12

n

5

5

5

5

5

5

5

5

5

                G: Group; F: Female; R: Recovery; SD: Standard Deviation; n:number of animals;*: Statistically significant (p<0.05)

 

 

TABLE 11 (Contd..,).  SUMMARY OF HAEMATOLOGY RECORD

 

Group, Sex & Dose

(mg/kg body weight/day)

Reticulocyte Count

Neutrophils

Lymphocytes

Monocytes

Eosinophils

Basophils

Absolute Reticulocyte Count

(Retic)

(Neut)

(Lymph)

(Mono)

(Eos)

(Baso)

(Retic)

(%)

(%)

(%)

(%)

(%)

(%)

(109cells/L)

G1, F & 0

Mean

1.61

19.76

75.22

2.46

0.84

0.46

110.56

±SD

0.31

2.55

1.87

0.61

0.35

0.13

20.46

n

5

5

5

5

5

5

5

G2, F & 100

Mean

1.58

24.28

70.00

3.40

0.56

0.42

106.28

±SD

0.57

9.05

9.06

0.89

0.25

0.19

38.40

n

5

5

5

5

5

5

5

G3, F & 300

Mean

1.22

21.50

73.50

2.48

0.88

0.38

78.84

±SD

0.41

6.95

7.64

0.80

0.64

0.25

29.53

n

5

5

5

5

5

5

5

G4, F & 1000

Mean

1.71

35.84*

56.92*

5.20*

0.52

0.48

120.26

±SD

0.52

8.33

9.49

1.89

0.08

0.16

36.41

n

5

5

5

5

5

5

5

G1R, F & 0

Mean

1.65

18.14

76.88

2.48

0.98

0.44

111.38

±SD

0.47

4.38

5.31

0.65

0.24

0.11

30.61

n

5

5

5

5

5

5

5

G4R, F & 1000

Mean

1.89

21.12

73.92

2.56

1.02

0.44

126.46

±SD

1.01

5.07

5.53

0.65

0.26

0.09

65.74

n

5

5

5

5

5

5

5

               G: Group; F: Female; R: Recovery; SD: Standard Deviation; n:number of animals; *: Statistically significant (p<0.05)

 

 

 

 

TABLE 11 (Contd..,).  SUMMARY OF HAEMATOLOGY RECORD

 

Group, Sex & Dose

(mg/kg body weight/day)

Absolute Neutrophils

Absolute Lymphocytes

Absolute Monocytes

Absolute Eosinophils

Absolute Basophils

Prothrombin Time

Activated Prothrombin Time

(Neut)

(Lymph)

(Mono)

(Eos)

(Baso)

(PT)

(APTT)

(103cells/µL)

(103cells/µL)

(103cells/µL)

(103cells/µL)

(103cells/µL)

(Seconds)

(Seconds)

G1, F & 0

Mean

1.72

6.65

0.22

0.07

0.05

19.28

23.44

±SD

0.76

2.88

0.09

0.06

0.03

1.81

2.31

n

5

5

5

5

5

5

5

G2, F & 100

Mean

2.19

6.32

0.31

0.05

0.04

18.70

19.72

±SD

0.86

1.36

0.10

0.03

0.02

0.53

1.77

n

5

5

5

5

5

5

5

G3, F & 300

Mean

1.69

6.66

0.20

0.06

0.04

19.94

26.46

±SD

0.57

4.03

0.07

0.03

0.05

3.60

5.13

n

5

5

5

5

5

5

5

G4, F & 1000

Mean

4.44*

6.74

0.65*

0.06

0.06

18.46

26.38

±SD

1.83

1.61

0.37

0.02

0.02

1.68

2.67

n

5

5

5

5

5

5

5

G1R, F & 0

Mean

1.50

6.84

0.21

0.08

0.04

19.68

28.84

±SD

0.28

2.83

0.05

0.03

0.01

3.05

5.11

n

5

5

5

5

5

5

5

G4R, F & 1000

Mean

2.02

7.20

0.24

0.10

0.04

27.44

21.28

±SD

0.47

1.57

0.03

0.04

0.02

7.74

6.50

n

5

5

5

5

5

5

5

               G: Group; F: Female; R: Recovery; SD: Standard Deviation; n:number of animals;*: Statistically significant (p<0.05)

 


 

TABLE 12.    SUMMARY OF CLINICAL CHEMISTRYRECORD

 

Group, Sex & Dose

(mg/kg body weight/day)

Glucose

Urea

Creatinine

Total Cholesterol

Triglycerides

Total Bilirubin

Total Protein

Albumin

Alanine aminotransferase

Aspartate aminotransferase

(GLU)

(CRE)

(CHO)

(TRI)

(BIT)

(TPR)

(ALB)

(ALT)

(AST)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

(g/dL)

(g/dL)

(U/L)

(U/L)

G1, M & 0

Mean

78.00

28.22

0.48

53.40

54.40

0.06

5.86

3.21

48.60

100.80

±SD

29.83

1.79

0.03

5.86

12.76

0.02

0.21

0.20

8.23

8.38

n

5

5

5

5

5

5

5

5

5

5

G2, M & 100

Mean

72.40

21.92*

0.47

42.40

55.00

0.05

6.16

3.25

50.20

105.40

±SD

8.79

3.56

0.03

14.31

8.75

0.03

0.46

0.20

8.14

7.60

n

5

5

5

5

5

5

5

5

5

5

G3, M & 300

Mean

52.80

21.54*

0.44*

49.60

60.80

0.04

5.98

3.10

55.60

112.60

±SD

9.52

3.25

0.01

7.57

16.77

0.03

0.30

0.10

7.44

18.28

n

5

5

5

5

5

5

5

5

5

5

G4, M & 1000

Mean

59.40

28.28

0.45

35.80*

54.60

0.09

5.66

3.11

68.00

136.60*

±SD

7.02

3.50

0.02

5.17

21.36

0.01

0.38

0.14

24.11

22.22

n

5

5

5

5

5

5

5

5

5

5

G1R, M & 0

Mean

90.20

27.82

0.60

44.80

44.20

0.02

6.50

3.17

42.40

75.40

±SD

7.19

2.53

0.05

9.09

5.97

0.00

0.14

0.09

1.95

9.91

n

5

5

5

5

5

5

5

5

5

5

G4R, M & 1000

Mean

88.60

28.86

0.60

49.40

51.60

0.03

6.48

3.16

45.40

86.00

±SD

10.01

2.13

0.02

7.92

11.28

0.01

0.32

0.10

7.23

8.89

n

5

5

5

5

5

5

5

5

5

5

         G: Group; M: Male; R: Recovery; SD: Standard Deviation; n:number of animals;*: Statistically significant (p<0.05)

 

 

 

 


TABLE 12 (Contd..,).  SUMMARY OF CLINICAL CHEMISTRY RECORD

 

Group, Sex & Dose

(mg/kg body weight/day)

Alkaline phosphatase

Calcium

Phosphorous

Cholinesterase

Globulin

Albumin/Globulin ratio

Blood Urea Nitrogen

Sodium

Potassium

Chloride

(ALP)

(CAL)

(PHO)

(CHE)

(GLO)

(A/G Ratio)

(BUN)

(Na)

(K)

(CLO)

(U/L)

(mg/dL)

(mg/dL)

(U/L)

(g/dL)

(mg/dL)

(mmol/L)

(mmol/L)

(mmol/L)

G1, M & 0

Mean

192.80

9.70

7.72

120.40

2.65

1.21

13.17

145.46

3.60

107.20

±SD

27.47

0.29

1.13

33.59

0.10

0.09

0.83

0.96

0.56

0.85

n

5

5

5

5

5

5

5

5

5

5

G2, M & 100

Mean

300.80

9.62

7.62

124.00

2.91

1.12

10.23*

147.06

3.36

107.80

±SD

121.48

0.28

0.13

25.72

0.27

0.06

1.66

1.90

0.14

1.73

n

5

5

5

5

5

5

5

5

5

5

G3, M & 300

Mean

409.00*

9.50

8.42

107.60

2.88

1.08*

10.05*

146.90

3.69

108.28

±SD

90.27

0.12

0.64

26.67

0.23

0.06

1.52

1.18

0.27

1.89

n

5

5

5

5

5

5

5

5

5

5

G4, M & 1000

Mean

229.80

8.94*

7.46

136.40

2.55

1.23

13.20

147.30

3.46

109.52

±SD

81.87

0.52

0.76

39.82

0.25

0.08

1.63

1.92

0.39

2.14

n

5

5

5

5

5

5

5

5

5

5

G1R, M & 0

Mean

149.60

10.02

6.96

173.80

3.33

0.96

12.98

142.24

3.34

106.58

±SD

37.75

0.18

0.23

18.32

0.16

0.06

1.18

1.19

0.33

0.90

n

5

5

5

5

5

5

5

5

5

5

G4R, M & 1000

Mean

168.40

9.90

7.32

167.00

3.32

0.96

13.47

142.68

3.44

106.72

±SD

30.36

0.14

0.28

20.92

0.26

0.07

0.99

0.95

0.52

1.62

n

5

5

5

5

5

5

5

5

5

5

  G: Group; M: Male; R: Recovery; SD: Standard Deviation; n:number of animals;*: Statistically significant (p<0.05)

 


 

TABLE 12 (Contd..,).  SUMMARY OF CLINICAL CHEMISTRY RECORD

 

Group, Sex & Dose

(mg/kg body weight/day)

Glucose

Urea

Creatinine

Total Cholesterol

Triglycerides

Total Bilirubin

Total Protein

Albumin

Alanine aminotransferase

Aspartate aminotransferase

(GLU)

(CRE)

(CHO)

(TRI)

(BIT)

(TPR)

(ALB)

(ALT)

(AST)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

(mg/dL)

(g/dL)

(g/dL)

(U/L)

(U/L)

G1, F & 0

Mean

72.60

26.48

0.50

55.20

41.20

0.06

6.40

3.33

37.40

104.20

±SD

11.93

2.64

0.02

4.71

4.15

0.01

0.32

0.20

6.07

15.43

n

5

5

5

5

5

5

5

5

5

5

G2, F & 100

Mean

80.80

23.94

0.49

56.20

39.40

0.07

6.52

3.38

46.40

101.60

±SD

21.43

4.09

0.02

7.53

8.50

0.02

0.18

0.11

5.86

12.60

n

5

5

5

5

5

5

5

5

5

5

G3, F & 300

Mean

63.20

25.76

0.49

48.20

39.60

0.09

6.28

3.38

50.20

104.20

±SD

12.07

2.66

0.03

12.30

7.50

0.02

0.34

0.19

11.65

15.67

n

5

5

5

5

5

5

5

5

5

5

G4, F & 1000

Mean

62.80

27.80

0.49

26.20*

35.80

0.13*

5.82*

3.19

59.60*

123.00

±SD

7.53

4.18

0.05

4.97

6.91

0.04

0.39

0.25

17.81

10.20

n

5

5

5

5

5

5

5

5

5

5

G1R, F & 0

Mean

100.40

35.60

0.70

63.40

43.00

0.02

7.06

3.57

34.00

74.60

±SD

11.24

3.43

0.03

10.67

4.00

0.01

0.30

0.18

2.55

6.35

n

5

5

5

5

5

5

5

5

5

5

G4R, F & 1000

Mean

112.00

38.52

0.67

53.60

44.80

0.02

7.02

3.46

36.20

74.20

±SD

22.51

4.71

0.05

9.56

15.17

0.00

0.33

0.26

7.05

10.69

n

5

5

5

5

5

5

5

5

5

5

         G: Group; F: Female; R: Recovery; SD: Standard Deviation; n:number of animals;*: Statistically significant (p<0.05)

 

 

 

 


TABLE 12 (Contd..,).  SUMMARY OF CLINICAL CHEMISTRY RECORD

 

Group, Sex & Dose

(mg/kg body weight/day)

Alkaline phosphatase

Calcium

Phosphorous

Cholinesterase

Globulin

Albumin/Globulin ratio

Blood Urea Nitrogen

Sodium

Potassium

Chloride

(ALP)

(CAL)

(PHO)

(CHE)

(GLO)

(A/G Ratio)

(BUN)

(Na)

(K)

(CLO)

(U/L)

(mg/dL)

(mg/dL)

(U/L)

(g/dL)

(mg/dL)

(mmol/L)

(mmol/L)

(mmol/L)

G1, F & 0

Mean

90.80

9.64

6.72

584.80

3.07

1.09

12.36

147.58

3.21

109.32

±SD

14.52

0.15

0.63

113.06

0.21

0.08

1.24

1.10

0.32

1.39

n

5

5

5

5

5

5

5

5

5

5

G2, F & 100

Mean

155.60*

9.80

7.04

420.00

3.14

1.08

11.17

147.62

3.31

108.40

±SD

24.54

0.42

1.19

98.38

0.19

0.09

1.91

1.19

0.35

1.37

n

5

5

5

5

5

5

5

5

5

5

G3, F & 300

Mean

201.20*

9.46

6.84

489.00

2.90

1.16

12.02

147.36

3.12

107.94

±SD

39.75

0.42

0.47

114.72

0.16

0.04

1.24

0.93

0.26

1.82

n

5

5

5

5

5

5

5

5

5

5

G4, F & 1000

Mean

191.00*

9.20

6.40

293.60*

2.63*

1.21

12.97

145.62

3.16

107.96

±SD

40.32

0.25

0.54

158.83

0.23

0.12

1.95

1.95

0.39

2.37

n

5

5

5

5

5

5

5

5

5

5

G1R, F & 0

Mean

88.40

10.16

6.06

746.00

3.49

1.02

16.61

142.98

3.31

107.54

±SD

26.50

0.23

0.45

147.97

0.14

0.03

1.60

0.83

0.08

0.59

n

5

5

5

5

5

5

5

5

5

5

G4R, F & 1000

Mean

101.40

10.02

6.14

607.80

3.56

0.97

17.98

142.82

3.63

106.44

±SD

24.93

0.40

0.30

414.52

0.12

0.07

2.20

0.79

0.50

0.92

n

5

5

5

5

5

5

5

5

5

5

 G: Group; F: Female; R: Recovery; SD: Standard Deviation; n:number of animals;*: Statistically significant (p<0.05)

 

 


TABLE 13.    SUMMARY OF URINE ANALYSIS RECORD

 

 

Group & Sex

G1 & M

G2 & M

G3 & M

G4 & M

G1R & M

G4R & M

Dose(mg/kg body weight/day)

0

100

300

1000

0

1000

Number of animals

5

5

5

5

5

5

Physical

Colour

Pale Yellow

2

4

4

5

4

5

Yellow

3

1

1

-

1

-

Appearance

Clear

3

3

4

5

4

5

Turbid

2

2

1

-

1

-

Volume (mL)

Mean

±SD

6.4

6.2

5.7

6.0

5.4

5.8

1.5

0.9

1.0

1.4

0.4

0.9

Chemical

pH

Mean

±SD

8.0

7.0

7.1

7.2

8.2

8.0

0.7

0.5

0.5

0.8

0.3

0.5

Specific gravity

Mean

±SD

1.011

1.011

1.014

1.010

1.010

1.009

0.005

0.005

0.009

0.000

0.000

0.002

Urobilinogen (mg/dL)

Mean

±SD

0.4

0.2

0.2

0.2

0.2

0.2

0.4

0.0

0.0

0.0

0.0

0.0

Bilirubin (mg/dL)

Neg

5

5

5

5

5

5

Ketones (mg/dL)

5

3

3

1

2

5

-

Neg

2

2

4

3

-

5

Blood (Ery/µL)

 

Neg

3

-

2

-

1

5

Ca10

1

1

1

2

-

-

Ca25

1

4

2

3

3

-

Ca80

-

-

-

-

1

-

Proteins (mg/dL)

Trace

1

1

3

2

4

1

Neg

1

2

1

3

-

4

30

1

2

1

-

-

-

100

1

-

-

-

1

-

>=300

1

-

-

-

-

-

Nitrite

Neg

4

5

1

3

5

5

Pos

1

-

4

2

-

-

Leucocytes (Leu/µL)

Neg

3

5

5

5

4

4

Ca15

2

-

-

-

1

1

Glucose (mg/dL)

Neg

5

5

5

5

5

5

Microscopic

Epithelial cells

0

4

4

3

4

3

3

0-1

1

-

2

1

1

1

1-2

-

1

-

-

1

1

Casts

Absent

5

5

5

5

5

5

Crystals

Present

5

5

5

5

5

5

             G: Group; M: Male; R: Recovery; Ca: Calculated approximately; SD: Standard Deviation;

               -: No observations noted;

 

 

 

 

 

TABLE 13 (Contd..,). SUMMARY OF URINE ANALYSIS RECORD

 

 

 

Group & Sex

G1 & F

G2 & F

G3 & F

G4 & F

G1R & F

G4R & F

Dose(mg/kg body weight/day)

0

100

300

1000

0

1000

Number of animals

5

5

5

5

5

5

Physical

Colour

Pale Yellow

3

5

-

3

2

3

Yellow

2

-

5

2

3

2

Appearance

Clear

4

4

2

3

2

3

Turbid

1

1

3

2

3

2

Volume (mL)

Mean

±SD

5.9

6.2

6.5

5.6

5.6

5.3

1.9

1.2

1.1

1.5

1.2

1.1

Chemical

pH

Mean

±SD

7.9

8.1

7.1

7.8

8.5

8.5

0.4

0.2

0.9

0.8

0.0

0.0

Specific gravity

Mean

±SD

1.011

1.008

1.018

1.012

1.006

1.009

0.007

0.003

0.010

0.008

0.002

0.004

Urobilinogen (mg/dL)

Mean

±SD

0.2

0.2

0.2

0.2

0.5

0.2

0.0

0.0

0.0

0.0

0.4

0.0

Bilirubin (mg/dL)

Neg

5

5

5

5

5

5

Ketones (mg/dL)

5

-

-

4

1

4

5

Neg

5

5

1

4

1

-

Blood (Ery/µL)

 

Neg

2

1

3

4

5

3

Ca10

-

1

1

1

-

1

Ca25

3

3

1

-

-

1

Proteins (mg/dL)

Trace

3

5

-

3

-

1

30

1

-

4

2

2

1

100

1

-

1

-

-

3

>=300

-

-

-

-

3

-

Nitrite

Neg

3

3

1

2

1

5

Pos

2

2

4

3

4

-

Leucocytes (Leu/µL)

Neg

4

5

4

5

-

2

Ca15

1

-

1

-

3

2

Ca70

-

-

-

-

-

1

Ca125

-

-

-

-

2

-

Glucose (mg/dL)

Neg

5

5

5

5

5

5

Microscopic

Epithelial cells

0

2

4

2

2

3

3

0-1

2

1

1

2

1

1

1-2

1

-

1

1

-

1

2-3

-

-

1

-

1

-

Casts

Absent

5

5

5

5

5

5

Crystals

Present

5

5

5

5

5

5

               G: Group; F: Female; R: Recovery; Ca: Calculated approximately; SD: Standard Deviation;

                 -: No observations noted.

          

 


TABLE 14.    SUMMARY OF ABSOLUTE ORGAN WEIGHT (g)RECORD

 

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

Adrenals

Thymus

Spleen

Epididymis

Testes

Heart

Kidneys

Brain

Liver

Prostate+Seminal vesicles with coagulating glands (PSC)

G1, M & 0

Mean

0.0632

0.3659

0.4042

1.0439

2.7126

0.8938

2.0984

1.8804

7.9667

1.6624

±SD

0.0155

0.0929

0.0493

0.0149

0.1284

0.0321

0.0489

0.0680

0.5287

0.2292

n

5

5

5

5

5

5

5

5

5

5

G2, M & 100

Mean

0.0653

0.4366

0.5396*

1.1197

2.6714

1.0447*

2.5895*

1.9089

10.4089*

1.8938

±SD

0.0093

0.0707

0.1257

0.0585

0.2764

0.0886

0.2819

0.0777

1.8151

0.0731

n

5

5

5

5

5

5

5

5

5

5

G3, M & 300

Mean

0.0650

0.5026

0.5181

1.1774*

3.0695*

1.1089*

2.9429*

2.0547

11.0130*

2.3908*

±SD

0.0095

0.0592

0.0359

0.0743

0.1480

0.0819

0.0495

0.2537

0.5488

0.5153

n

5

5

5

5

5

5

5

5

5

5

G4, M & 1000

Mean

0.0557

0.3316

0.4204

1.0225

2.8182

0.8916

2.2137

1.8334

9.6882

1.7699

±SD

0.0053

0.1217

0.0302

0.0683

0.1642

0.0402

0.2962

0.0606

1.5450

0.3296

n

5

5

5

5

5

5

5

5

5

5

G1R, M & 0

Mean

0.0467

0.3566

0.4846

1.2428

2.8844

1.1545

2.3934

1.9366

9.6305

2.8592

±SD

0.0037

0.1127

0.0726

0.0746

0.1545

0.0923

0.3351

0.0947

0.8507

0.3037

n

5

5

5

5

5

5

5

5

5

5

G4R, M & 1000

Mean

0.0569

0.3963

0.5472

1.2226

3.0443

1.2380

2.7301

2.0031

10.0676

2.7223

±SD

0.0098

0.0409

0.0790

0.0690

0.0783

0.0834

0.4558

0.0691

1.0711

0.2980

n

5

5

5

5

5

5

5

5

5

5

G: Group; M: Male; R: Recovery; SD: Standard Deviation; n:number of animals;*: Statistically significant (p<0.05)

 

 


TABLE 14 (Contd..,).  SUMMARY OF ABSOLUTE ORGAN WEIGHT (g) RECORD

 

Group, Sex & Dose

(mg/kg body weight/day)

Adrenals

Thymus

Spleen

Uterus

Ovaries

Heart

Kidneys

Brain

Liver

G1, F & 0

Mean

0.0687

0.4911

0.4541

0.4964

0.1554

0.8303

1.7937

1.8833

7.4558

±SD

0.0077

0.0901

0.0832

0.1060

0.0191

0.1074

0.3057

0.0928

1.3075

n

5

5

5

5

5

5

5

5

5

G2, F & 100

Mean

0.0802

0.4696

0.4551

0.5508

0.1450

0.8371

1.7849

1.7964

7.9453

±SD

0.0080

0.0815

0.0193

0.1484

0.0260

0.0593

0.1003

0.1504

0.1984

n

5

5

5

5

5

5

5

5

5

G3, F & 300

Mean

0.0689

0.4435

0.4854

0.5317

0.1554

0.8306

1.9649

1.9528

8.6017

±SD

0.0101

0.1219

0.0949

0.1248

0.0197

0.0454

0.0877

0.0634

0.4803

n

5

5

5

5

5

5

5

5

5

G4, F & 1000

Mean

0.0803

0.5359

0.5362

0.5131

0.1453

0.8156

2.0959*

1.9882

9.2202*

±SD

0.0154

0.1220

0.1095

0.0434

0.0238

0.1035

0.1080

0.0863

0.3073

n

5

5

5

5

5

5

5

5

5

G1R, F & 0

Mean

0.0590

0.3688

0.4692

0.6375

0.1525

0.8816

1.7360

1.9808

7.2510

±SD

0.0107

0.1099

0.1049

0.1581

0.0252

0.0700

0.1980

0.0704

0.5332

n

5

5

5

5

5

5

5

5

5

G4R, F & 1000

Mean

0.0789*

0.3649

0.4532

0.5133

0.1533

1.0073

1.7925

1.8895

8.0583

±SD

0.0111

0.0650

0.0937

0.1131

0.0257

0.3164

0.1536

0.1411

1.0192

n

5

5

5

5

5

5

5

5

5

G: Group; F: Female; R: Recovery; SD: Standard Deviation; n:number of animals;*: Statistically significant (p<0.05)

       

 


 

TABLE 15.   SUMMARY OF ORGAN WEIGHT RELATIVE TO FASTING BODY WEIGHT(%)RECORD

 

Group, Sex & Dose

(mg/kg body weight/day)

Fasting Body Weight (g)

Adrenals

Thymus

Spleen

Epididymis

Testes

Heart

Kidneys

Brain

Liver

Prostate+Seminal vesicles with coagulating glands (PSC)

G1, M & 0

Mean

266.88

0.0241

0.1390

0.1527

0.3941

1.0233

0.3380

0.7919

0.7112

3.0142

0.6302

±SD

26.22

0.0074

0.0429

0.0250

0.0386

0.1006

0.0410

0.0734

0.0864

0.4135

0.1227

n

5

5

5

5

5

5

5

5

5

5

5

G2, M & 100

Mean

269.28

0.0244

0.1616

0.1987*

0.4168

0.9969

0.3886

0.9621*

0.7129

3.8483*

0.7074

±SD

21.68

0.0044

0.0182

0.0316

0.0191

0.1282

0.0275

0.0845

0.0683

0.4465

0.0700

n

5

5

5

5

5

5

5

5

5

5

5

G3, M & 300

Mean

276.03

0.0236

0.1817

0.1884

0.4271

1.1139

0.4035*

1.0700*

0.7488

3.9980*

0.8656*

±SD

16.65

0.0040

0.0129

0.0189

0.0263

0.0608

0.0463

0.0801

0.1222

0.2607

0.1828

n

5

5

5

5

5

5

5

5

5

5

5

G4, M & 1000

Mean

228.95*

0.0243

0.1446

0.1840

0.4477*

1.2354*

0.3901

0.9643*

0.8038

4.2342*

0.7731

±SD

13.09

0.0010

0.0515

0.0152

0.0382

0.1185

0.0217

0.0830

0.0670

0.6522

0.1370

n

5

5

5

5

5

5

5

5

5

5

5

G1R, M & 0

Mean

321.27

0.0148

0.1134

0.1514

0.3918

0.9080

0.3620

0.7477

0.6088

3.0095

0.8953

±SD

38.28

0.0026

0.0425

0.0195

0.0558

0.1161

0.0356

0.0842

0.0699

0.1788

0.1008

n

5

5

5

5

5

5

5

5

5

5

5

G4R, M & 1000

Mean

307.69

0.0184

0.1286

0.1780

0.3980

0.9921

0.4033

0.8841

0.6536

3.2694

0.8859

±SD

18.09

0.0026

0.0080

0.0248

0.0235

0.0629

0.0352

0.1107

0.0566

0.2560

0.0950

n

5

5

5

5

5

5

5

5

5

5

5

  G: Group;M: Male;R: Recovery; SD: Standard Deviation; n: Number of animals;*: Statistically significant (p<0.05)


TABLE 15 (Contd..,).  SUMMARY OF ORGAN WEIGHT RELATIVE TO FASTING BODY WEIGHT (%) RECORD

 

Group, Sex & Dose

(mg/kg body weight/day)

Fasting Body Weight (g)

Adrenals

Thymus

Spleen

Uterus

Ovaries

Heart

Kidneys

Brain

Liver

G1, F & 0

Mean

206.98

0.0333

0.2371

0.2183

0.2399

0.0758

0.4005

0.8635

0.9150

3.5821

±SD

18.48

0.0036

0.0367

0.0252

0.0495

0.0136

0.0292

0.0861

0.0841

0.3018

n

5

5

5

5

5

5

5

5

5

5

G2, F & 100

Mean

206.48

0.0389

0.2281

0.2204

0.2651

0.0705

0.4051

0.8646

0.8694

3.8530

±SD

8.63

0.0034

0.0420

0.0044

0.0596

0.0143

0.0141

0.0401

0.0537

0.1769

n

5

5

5

5

5

5

5

5

5

5

G3, F & 300

Mean

209.10

0.0331

0.2110

0.2313

0.2536

0.0745

0.3971

0.9407

0.9344

4.1201*

±SD

7.03

0.0054

0.0525

0.0388

0.0558

0.0104

0.0151

0.0547

0.0325

0.3113

n

5

5

5

5

5

5

5

5

5

5

G4, F & 1000

Mean

202.34

0.0399

0.2678

0.2648

0.2543

0.0719

0.4029

1.0393*

0.9854

4.5709*

±SD

14.44

0.0083

0.0746

0.0485

0.0229

0.0118

0.0396

0.0788

0.0611

0.2768

n

5

5

5

5

5

5

5

5

5

5

G1R, F & 0

Mean

231.70

0.0256

0.1578

0.2014

0.2743

0.0657

0.3805

0.7485

0.8564

3.1304

±SD

10.00

0.0053

0.0406

0.0376

0.0619

0.0090

0.0255

0.0677

0.0539

0.2156

n

5

5

5

5

5

5

5

5

5

5

G4R, F & 1000

Mean

231.78

0.0340*

0.1591

0.1959

0.2205

0.0662

0.4384

0.7776

0.8158

3.4799

±SD

15.01

0.0040

0.0358

0.0419

0.0407

0.0108

0.1540

0.0997

0.0446

0.3997

n

5

5

5

5

5

5

5

5

5

5

G: Group; F: Female; R: Recovery; SD: Standard Deviation; n:number of animals;*: Statistically significant (p<0.05)

      


TABLE 16.SUMMARY OF GROSS PATHOLOGY FINDINGS

Organs/ Lesions

Group

G1

G1R

G2

G3

G4

G4R

Dose (mg/kg body  

        weight/day)

0

0

100

300

1000

1000

Sex

M

F

M

F

M

F

M

F

M

F

M

F

No. of Animals

5

5

5

5

5

5

5

5

5

5

5

5

No Abnormality Detected

5

5

5

5

5

5

5

5

5

5

5

5

The numeral indicates number of animal/s; M: Male; F: Female; G: Group.

 


TABLE 17.SUMMARY OFHISTOPATHOLOGY FINDINGS

Sex

Male

Female

Group

G1

G4

G1

G4

Dosage (mg/kg body weight/day)

0

1000

0

1000

Liver

5

5

5

5

MNC infiltration:Minimal

0

0

1

1

Kidneys

5

5

5

5

Cortex; tubular basophilia; unilateral : Minimal

1

0

0

1

Lungs

5

5

5

5

Perivascular MNC infiltration: Minimal

1

1

0

0

Thyroid

5

5

5

5

Ectopic thymus

1

0

0

1

Note: The numeral indicates number of animal/s with lesion; G: group; MNC: Mononuclear cell.

 

Conclusions:
Based on the observed results, the No Observed Adverse Effect Level (NOAEL) of test item Hostacor ITD is 1000 mg/kg/day when administered for a period of 28 consecutive days by oral (gavage) route to Sprague Dawley under the test conditions and doses employed.
Executive summary:

The objective of this studywas to assess the toxic potential of the test item Hostacor ITD when administered for a period of 28 consecutive days repeatedly by oral (gavage) to Sprague Dawley rats. This studyprovidesinformation on major toxic effects, target organs, possibility of cumulative effects and also to assess the reversibility of effects (after 14 days recovery period) and an estimate of the No Observed Adverse Effect Level (NOAEL). This study wasperformed in accordance with OECD Guideline No. 407, “Repeated Dose 28-Day Oral Toxicity Study in Rodents” adopted on 03 October 2008.

A total of 60 Sprague Dawleyrats(30males +30females)were distributed to4mainand2recovery groups. Each main groupand recovery groupconsisted of 5animals/sex/group.The test item was formulated using corn oil.The animals allocated togroupsG2, G3 and G4/G4Rwere administeredwithoral dosesof100, 300 and 1000 mg/kg body weight/day of test item, respectivelythrough oral gavage.The animals allocated togroupG1/G1Rreceived vehicle corn oil. The vehicle/test item formulations were administered at an equivolume of 10 mL/kg.

The dose formulations wereanalyzed duringweek 1 andweek 4of the treatment period and the results indicated that the obtained concentrationswere within the acceptable range of 85 to 115%of the nominal concentrationwith less than 10% RelativeStandardDeviation.

All animals were observed once daily for clinical signs and twice daily for mortality; weekly for detailed clinical examination, body weight and feed consumption; ophthalmoscopic examination was performed during acclimatization and during    week4for main groups (G1andG4) and during week6for recovery groups (G1RandG4R). Functional observational battery was performed during week4forallmain groupanimalsand during week6for recovery groupanimals(G1RandG4R).

At the end of treatment and recovery period (day 29 for main group and day 43 for recovery group), blood and urine samples were collected and analyzed. Subsequently, the animals were sacrificed and subjected to gross pathological examination and the organs were collected and subjected to histopathological examination.

Norelevantclinical signs of toxicity was observed in any of the vehicle or test item administered animals up tothe high dosegroup. Aslight wet perineum was observed ina fewhigh dose test item administered animals (males: 2/5 and females: 2/5 in main group animals; males: 1/5 and females: 2/5 in recovery group animals)which is not considered to be of toxicological relevance.

No treatment related changes in body weight, percent change in body weight with respect to day 1, feed consumption and ophthalmoscopic examination were noted. No adverse treatment related changes were noted in haematology, clinical chemistry and urinalysis parameters. No toxicologically significant changes were noted in organ weights and its ratios.

No treatment related gross and histopathology changes were noted in any organ and/or tissue.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
high quality, GLP sudies according to OECD Test Guideline

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

No adverse effects are observed in limit tests (up to 1000 mg/kg of body weight) according to OECD TG 407.

Justification for classification or non-classification

No relevant toxic effects were observed in oral repeated dose toxicity studies in rats (highest dose tested 1000 mg/kg bw/day) in OECD 407 and 14-day dose range findings studies performed with the submission substance or surrogates). No data on repeated dose toxicity after inhalation or dermal exposure are available. Therefore the submission substance does not have to be classified for specific target organ toxicity – repeated exposure or repeated dose toxicity according to Regulation (EC) No 1272/2008 or to Council Directive 67/548/EEC.