Registration Dossier

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Sept 2017 - May 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Animal Breeding, RCC Laboratories India Private Limited, Genome Valley, Turkapally, Shameerpet (Mandal), Ranga Reddy District, Hyderabad -500 078 India.
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 7-8 weeks
- Weight at study initiation: Male: 164.5 to 198.9 g, Female: 132.6 to 158.2 g
- Housing: In groups of two animals of same sex in Polycarbonate cages (Approximate internal dimensions of 365 mm x 202 mm x 180 mm height) with corncob bedding.
- Diet (e.g. ad libitum): Teklad Certified Global 14% Protein Rodent Maintenance Diet (Lot No. 2014C-082417MA) from ENVIGO was provided ad libitum.
- Water (e.g. ad libitum): Aquaguard filtered tap water was provided ad libitum.
- Acclimation period: 9 days

DETAILS OF FOOD AND WATER QUALITY:
Results of analyses for contaminants are archived at RCC Laboratories India Private

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20.9 to 22.8°C
- Humidity (%): 57 to 65%
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES:
From: 4.25. Sept. 2017 To: 1. Feb. 2018

Route of administration:

oral: gavage

Details on route of administration:

The test item was administered to the test system, for the period of 90 consecutive days by oral route using oral gavage needle.

Vehicle:

corn oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:

VEHICLE
- Justification for use and choice of vehicle (if other than water): based on preliminary solubility testing which was performed in the test facility before the study initiation date
- Concentration in vehicle: 5 mg/mL, 20 mg/mL, 40 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Gas chromotography
Samples of the dose formulations were taken immediately after dilution of the test item with the diluent (vehicle) on treatment start date and last week of treatment for homogeneity (mean of homogeneity was given as dose concentration) analyses. On week 7, samples of all dose formulations were assayed for the applied dose in triplicates. The method was developed and validated prior to analysis. The method was either matrix assisted validation or external standard method of validation. The validation and analysis was carried out as per SANCO/3029/99 rev.4.
The test item in corn oil was assayed on three occasions
01 Stability of test item at 0th, 2nd and 4th hour in corn oil matrix for the fortified concentrations of 5 mg/mL and 40 mg/mL.
02 Initial homogeneity test on Top, mid and bottom layers of corn oil fortified with of test item Viz., 5 mg/mL (Low dose), 20 mg/mL (Intermediate dose), and 40 mg/mL (High dose)
03 Dose concentration of test item in corn oil fortified with 5 mg/mL (Low dose), 20 mg/mL (Intermediate dose), and 40 mg/mL (High dose).
04 Final homogeneity test on Top, mid and bottom layers of corn oil fortified with of test item Viz., 5 mg/mL (Low dose), 20 mg/mL (Intermediate dose), and 40 mg/mL (High dose). Calibration solution, CS5 was used as bracketing standard.

Duration of treatment / exposure:

90 days

Frequency of treatment:

daily

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: The doses to be employed for this study were selected, based on the 28 days repeated dose toxicity dose range finding study
- Post-exposure recovery period in satellite groups: 28 days

Positive control:

n.a.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
twice daily for mortality and on the first 3 days of treatment for clinical observations
once daily for clinical observations

DETAILED CLINICAL OBSERVATIONS: Yes
Once prior to the first exposure and weekly thereafter

BODY WEIGHT: Yes
once weekly

FOOD CONSUMPTION AND COMPOUND INTAKE:
once weekly

OPHTHALMOSCOPIC EXAMINATION: Yes
Prior to the administration of the test item (in 20% of total population) and during week 13 in all animals of group 1 and 4. animals during week 17. Tropicamide was used as a mydriatic agent.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: week 13 (main groups), week 17 (recovery groups)
- Anaesthetic used for blood collection: Yes, isoflurane inhalation anaesthesia
- Animals fasted: Yes
- How many animals: all animals
- Parameters checked:
erythrocyte count, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular concentration, platelet count, total leukocyte count, differential leukocyte count, reticulocytes, prothrombin time, activated partial thromboplastin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: week 13 (main groups), week 17 (recovery groups)
- Animals fasted: Yes
- How many animals: all animals
- Parameters checked:
glucose, urea, blood urea nirtogen, creatinine, cholesterol, triglycerides. aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma glutamyltransferase, bilirubin, sodium, potassium, chloride, protein total, albumin, globulin, A G ratio

URINALYSIS: Yes
- Time schedule for collection of urine: week 13 (main groups), week 17 (recovery groups)
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters checked:
volume, specific gravitiy, colour, clarity, pH, erythrocytes, leukocytes, bilirubin, urobiliongen, ketone bodies, proteins and glucose

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations:during week 12 and 17
- Dose groups that were examined: all
- Battery of functions tested: sensory activity / grip strength / motor activity

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
Necropsy: all animals
organ weight (all animals): brain ,heart, liver, thymus, kidneys, adrenals, spleen, testes, epididymides, oavaries, uterus, prostate and seminal vescles with coagulating glands

HISTOPATHOLOGY: Yes
in control and high dose animals (main groups)
addtionally organs that revealed changes in low, mid dose (main groups) and all recovery groups
Adrenal glands, Aorta, Bone marrow (Sternum), Brain (cerebrum, cerebellum, medulla/ pons), Cecum, Cervix, Colon, Duodenum, Epididymides, Eye, Heart, Ileum with Peyer's patches, Jejunum, Kidneys, Liver, Lungs (inflated with NBF at necropsy), Lymph nodes (mesenteric, axillary), Oesophagus, Ovaries, Pancreas, Peripheral nerve (sciatic or tibial), Pituitary, Prostate, Rectum, Salivary gland, Seminal vesical with coagulating glands, Skeletal muscle, Skin with Mammary gland, Spinal cord (cervical, midthoracic, lumbar), Spleen,stomach, Testes, Thymus, Thyroid with parathyroid, Trachea, Urinary bladder (inflated with NBF at necropsy), Uterus, Vagina, All Gross lesions

Statistics:

The following statistical methods were used to analyze the body weight, feed consumption, functional observation battery parameters,organ weights as well as clinical pathology data.
• Data was summarized in tabular form. Statistical analysis was performed using statplus program.
• All the data was checked for normality with Shapiro-Wilk W test
• Statistical significances of differences was calculated with one-way analysis of variance(ANOVA). Values were given as mean ± standard deviation (SD).
• t-test/ all ANOVA with group variables were used to compare the difference between treated and control groups.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

No test item related clinical signs of illness were observed in any of the animals during treatment in groups of control (G1), Low dose (G2), Intermediate dose (G3) and recovery period (G1R). However, Dullness was obesrved in high dose (G4) in one male and one female, similarly one male and two females exhibited dullness in high dose recovery (G4R).

Mortality:

no mortality observed

Description (incidence):

Animals treated with test item at dose levels of 50, 200 and 400 mg/kg bw/d did not show any mortality in both males and females during the experimental period of 90 days and the recovery period.

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

The body weight of male animals of all treatment groups were comparable with the control group animals during the dosing period and recovery period.
In females, significant decreases in body weight gain were observed for (G4) on day 85 when compared to the vehicle treated animals. The body weight gain of female animals of recovery high dose group was comparable with the recovery control group animals during the dosing period and recovery period.
The changes observed were considered to be as biological variation commonly recorded for this species, strain and age.

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

During the 90 consecutive days of dosing period, the quantity of feed consumed by animals across different dose groups was found to be no significantly different as compared with the control animals.

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

Males: Significant increase of NEU and decrease of lymph in intermediate dose (G3) and high dose (G4) as compared with control dose (G1). Significant increase of mono in high (G4), intermediate dose (G3) low dose (G2) as compared with control group (G1). Significant decrease of EOS in low dose (G2) as compared with control group (G1).
Females: There was significant increase in Neut and decrease in Lymph in (G4) as compared with (G1).

In recovery males, there was significant increase of PCV in G4R as compared with control recovery (G1R). In recovery females, there was significant decrease of white blood cell count (WBC) in female as compared with control recovery (G1R). The changes observed in the hematological parameters are marginal and could not be attributed to the test item administration as these values were within biological variation.

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

Males: Significant increase of ALP and AST in intermediate (G3) and high (G4) dose as compared with control (G1) group. Significant increase of ALT in high (G4) dose as compared with control (G1).
Females: there was significant increase of AST in intermediate (G3) as compared with control (G1). Significant increase of CREA in intermediate (G3) dose as compared with (G1) dose.
In recovery males, there was significant decrease in ALP (G4R) when compared with control recovery (G1R). In recovery females, there was significant increase in UREA, BUN, CREA and ALB in (G4R) when compared with control recovery (G1R). The changes observed in the clinical biochemistry parameters are marginal and could not be attributed to the test item administration as these values were within biological variation.

Urinalysis findings:

effects observed, non-treatment-related

Description (incidence and severity):

In males, there was significant decrease of volume in high (G4) dose as compared with control (G1). In females there was significant increase of SG in intermediate (G3) ) and low (G2) as compared with control (G1). The changes observed in the urine analysis parameters are marginal and could not be attributed to the test item administration as these values were within biological variation.

Behaviour (functional findings):

no effects observed

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

There was no significance changes observed in absolute and relative organ weights of treatment and recovery groups. In relative organ weights of males, there was significant increase of liver in high (G4) dose as compared with control (G1).
Despite this macroscopical difference, no test item related change was observed in these organs from treatment groups. Hence, the changes observed in organ weight were considered to be as biological variation.

Gross pathological findings:

no effects observed

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Treatment related histopathological findings included chronic inflammation in the non glandular region of stomach observed in treated males and females. Additionally hyperplasia and hyperkeratosis of squamous epithelium of non glandular stomach was also observed in a few animals in these groups. There was single incidence of erosion in glandular stomach in one male treated at 400 mg/kg bw. Chronic inflammation in non glandular stomach consisted of infiltration in lamina propria and/or submucosa with mostly mononuclear cells and occasional polymorphonuclear cells. Edema or/and necrosis of the squamous epithelium was observed in a few cases. The distribution of the lesion was focal or multifocal. The lesion in non glandular stomach was observed in all the treated groups. The severity of the lesion was minimal to mild in all the treated groups. . The incidence and severity of the lesion in females treated at 400mg/kg was higher when compared to males treated with the same dose. The lesions observed in the forestomach may be attributed to direct local effects of the test item administered orally. In the recovery high dose groups, all the stomach tissues were within normal histological limits indicating complete recovery. The lesion was considered non adverse as the incidence and severity of the lesion was low and complete recovery was at the end of 28 day recovery period.

Incidence of stomach effects
- Males:
Inflammation: 0/10 (0 mg/kg bw/d), 4/10 (50 mg/kg bw/d), 4/10 (200 mg/kg bw/d), 2/10 (400 mg/kg bw/d), 0/10 (0 mg/kg bw/d recovery group), 0/10 (400 mg/kg bw/d recovery group)
Hyperpalsia/Hyperkeratosis: 0/10 (0 mg/kg bw/d), 1/10 (50 mg/kg bw/d), 1/10 (200 mg/kg bw/d), 2/10 (400 mg/kg bw/d), 0/10 (0 mg/kg bw/d recovery group), 0/10 (400 mg/kg bw/d recovery group)
Erosion (glandular, focal): 0/10 (0 mg/kg bw/d), 0/10 (50 mg/kg bw/d), 0/10 (200 mg/kg bw/d), 1/10 (400 mg/kg bw/d), 0/10 (0 mg/kg bw/d recovery group), 0/10 (400 mg/kg bw/d recovery group)

- Females:
Inflammation: 0/10 (0 mg/kg bw/d), 3/10 (50 mg/kg bw/d), 4/10 (200 mg/kg bw/d), 6/10 (400 mg/kg bw/d), 0/10 (0 mg/kg bw/d recovery group), 0/10 (400 mg/kg bw/d recovery group)
Hyperpalsia/Hyperkeratosis: 0/10 (0 mg/kg bw/d), 0/10 (50 mg/kg bw/d), 2/10 (200 mg/kg bw/d), 6/10 (400 mg/kg bw/d), 0/10 (0 mg/kg bw/d recovery group), 0/10 (400 mg/kg bw/d recovery group)

Single incidence of microscopic findings observed in other organs in various groups were considered incidental.

Histopathological findings: neoplastic:

no effects observed

Other effects:

no effects observed

FEED CONSUMPTION (G/ANIMAL/DAY)

MALES

TREATMENT		Group 1	Group 2	Group 3	Group 4
Week 1	Mean	19.15	19.48	19.84	17.55
Week 1	SD	1.018	0.346	0.816	3.400
Week 2	Mean	20.26	18.31	17.31	20.25
Week 2	SD	3.435	1.397	2.147	1.541
Week 3	Mean	22.58	20.68	20.17	19.93
Week 3	SD	1.740	2.347	2.173	2.585
Week 4	Mean	20.18	19.87	16.46	17.54
Week 4	SD	1.448	3.768	1.238	2.397
Week 5	Mean	18.83	17.17	17.07	19.22
Week 5	SD	2.033	1.440	0.901	2.938
Week 6	Mean	17.42	15.25	15.42	17.36
Week 6	SD	1.546	0.801	1.323	4.212
Week 7	Mean	16.71	15.67	14.68	15.76
Week 7	SD	1.635	0.918	0.730	1.610
Week 8	Mean	16.79	16.20	16.49	15.79
Week 8	SD	1.423	1.852	1.205	1.844
Week 9	Mean	17.75	16.70	16.96	16.26
Week 9	SD	1.346	1.471	1.374	1.693
Week 10	Mean	16.11	15.99	15.24	14.87
Week 10	SD	1.202	1.483	0.617	1.380
Week 11	Mean	17.50	17.62	17.56	16.87
Week 11	SD	0.395	1.680	1.425	1.681
Week 12	Mean	18.18	18.72	18.55	17.41
Week 12	SD	1.619	1.219	1.073	2.447
Week 13	Mean	15.37	15.98	15.93	15.87
Week 13	SD	1.046	1.991	1.351	0.729

FEED CONSUMPTION (G/ANIMAL/DAY) (CONTD.)

MALES

Recovery (R)		Group 1R	Group 4R	Recovery (R)		Group 1R	Group 4R
Week 1	Mean	17.77	17.18	Week 10	Mean	17.99	17.06
Week 1	SD	2.256	2.757	Week 10	SD	1.000	0.700
Week 2	Mean	21.57	20.22	Week 11	Mean	17.78	17.36
Week 2	SD	1.711	1.194	Week 11	SD	1.175	0.916
Week 3	Mean	21.55	21.25	Week 12	Mean	18.98	18.70
Week 3	SD	2.132	2.176	Week 12	SD	0.373	0.563
Week 4	Mean	17.68	17.65	Week 13	Mean	19.28	17.99
Week 4	SD	1.097	1.107	Week 13	SD	1.405	1.265
Week 5	Mean	18.40	18.41	Week 14	Mean	19.41	19.22
Week 5	SD	1.468	1.364	Week 14	SD	0.238	0.959
Week 6	Mean	16.98	16.76	Week 15	Mean	19.43	19.13
Week 6	SD	0.822	0.691	Week 15	SD	0.227	0.455
Week 7	Mean	16.39	16.49	Week 16	Mean	19.52	19.24
Week 7	SD	3.537	1.029	Week 16	SD	0.245	0.293
Week 8	Mean	16.56	15.88	Week 17	Mean	21.44	21.79
Week 8	SD	0.927	0.834	Week 17	SD	0.174	1.147
Week 9	Mean	18.80	18.57
Week 9	SD	0.377	0.691

FEED CONSUMPTION (G/ANIMAL/DAY) (CONTD.)

FEMALES

TREATMENT		Group 1	Group 2	Group 3	Group 4
Week 1	Mean	13.28	12.57	11.93	10.70
Week 1	SD	0.631	1.286	2.569	2.459
Week 2	Mean	13.37	11.46	12.84	13.13
Week 2	SD	2.122	1.356	1.356	1.491
Week 3	Mean	16.69	13.41*	14.24	12.42*
Week 3	SD	1.260	2.296	1.679	1.635
Week 4	Mean	11.87	11.54	11.33	9.79^*#&
Week 4	SD	0.774	1.041	0.819	0.496
Week 5	Mean	12.06	11.56	12.35	12.10
Week 5	SD	0.694	1.102	1.081	1.580
Week 6	Mean	12.12	10.97	12.07	10.37
Week 6	SD	0.801	2.383	0.558	1.316
Week 7	Mean	11.88	10.67	11.90	11.76
Week 7	SD	0.650	1.193	0.658	2.780
Week 8	Mean	12.99	11.82	12.22	11.05
Week 8	SD	1.143	2.046	1.538	2.076
Week 9	Mean	13.07	12.14	12.81	11.58
Week 9	SD	1.206	1.391	1.240	1.147
Week 10	Mean	12.44	10.53	11.05	9.53
Week 10	SD	1.870	1.939	1.893	1.193
Week 11	Mean	13.93	11.39	13.50	10.21*
Week 11	SD	1.127	1.645	3.353	1.295
Week 12	Mean	15.57	11.99*	13.24	11.98*
Week 12	SD	1.686	0.726	0.916	1.597
Week 13	Mean	13.61	10.94	11.94	11.15
Week 13	SD	1.490	1.727	1.428	1.643

*-Significant at p ≤ 0.05 with Group 1, # - Significant at p ≤ 0.05 with Groups 2, & - Significant at p ≤ 0.05 with Group 3

FEED CONSUMPTION (G/ANIMAL/DAY) (CONTD.)

FEMALES

Recovery (R)		Group 1R	Group 4R	Recovery (R)		Group 1R	Group 4R
Week 1	Mean	11.36	10.65	Week 10	Mean	11.58	10.13
Week 1	SD	2.607	2.559	Week 10	SD	1.067	1.306
Week 2	Mean	15.15	13.57	Week 11	Mean	12.32	10.74
Week 2	SD	2.458	0.936	Week 11	SD	1.051	1.233
Week 3	Mean	15.43	14.77	Week 12	Mean	13.25	12.14
Week 3	SD	1.916	1.502	Week 12	SD	0.739	1.384
Week 4	Mean	12.65	12.36	Week 13	Mean	15.35	13.21
Week 4	SD	0.408	1.016	Week 13	SD	1.074	2.537
Week 5	Mean	12.69	12.99	Week 14	Mean	14.40	14.57
Week 5	SD	0.944	1.038	Week 14	SD	0.662	0.573
Week 6	Mean	11.39	10.82	Week 15	Mean	13.98	13.78
Week 6	SD	0.841	0.608	Week 15	SD	0.532	1.285
Week 7	Mean	11.95	11.65	Week 16	Mean	13.78	14.04
Week 7	SD	1.156	1.172	Week 16	SD	0.812	1.410
Week 8	Mean	11.15	12.25	Week 17	Mean	15.23	14.77
Week 8	SD	1.357	0.862	Week 17	SD	0.619	0.722
Week 9	Mean	13.56	12.39
Week 9	SD	2.170	1.407

1.1 BODY WEIGHTS (G)

MALES

TREATMENT		Group 1	Group 2	Group 3	Group 4
Day 1	Mean	178.0	181.4	182.4	179.0
	SD	5.28	3.16	7.30	4.88
	Min	170.5	178.4	172.9	171.4
	Max	187.3	189.0	198.9	188.8
	N	10	10	10	10
Day 8	Mean	211.2	209.4	217.0	214.5
	SD	10.91	10.20	8.69	11.50
	Min	197.2	190.7	206.6	196.1
	Max	229.3	223.4	234.3	229.7
	N	10	10	10	10
Day 15	Mean	240.2	237.5	239.9	236.6
	SD	15.56	8.32	8.93	18.79
	Min	219.8	224.9	227.5	215.1
	Max	270.1	254.2	252.5	267.0
	N	10	10	10	10
Day 22	Mean	264.4	257.6	259.4	253.6
	SD	18.55	8.62	11.09	22.02
	Min	246.5	243.3	237.6	224.5
	Max	302.2	273.2	273.4	292.3
	N	10	10	10	10
Day 29	Mean	288.6	276.5	277.1	273.8
	SD	19.44	11.87	13.94	25.40
	Min	265.0	254.6	249.3	238.7
	Max	325.9	301.2	296.4	320.7
	N	10	10	10	10

TREATMENT		Group 1	Group 2	Group 3	Group 4
Day 36	Mean	307.0	290.0	291.1	293.3
	SD	26.28	18.65	16.38	28.65
	Min	272.5	260.8	263.9	257.7
	Max	362.0	334.3	317.0	345.4
	N	10	10	10	10
Day 43	Mean	322.8	308.3	305.5	303.3
	SD	26.48	19.24	18.31	32.64
	Min	287.5	280.4	274.4	258.1
	Max	367.7	354.1	332.6	361.5
	N	10	10	10	10
Day 50	Mean	332.2	320.1	317.5	316.6
	SD	29.26	20.37	20.93	37.44
	Min	292.7	289.3	281.8	260.4
	Max	379.1	367.8	349.9	382.3
	N	10	10	10	10
Day 57	Mean	343.2	331.4	328.3	325.4
	SD	33.77	23.73	23.29	36.23
	Min	293.8	300.6	292.5	273.9
	Max	391.1	389.8	365.7	385.2
	N	10	10	10	10
Day 64	Mean	353.1	343.3	340.6	336.8
	SD	34.42	26.55	24.15	38.57
	Min	303.2	306.3	307.0	284.9
	Max	399.1	404.6	381.7	399.6
	N	10	10	10	10

TREATMENT		Group 1	Group 2	Group 3	Group 4
Day 71	Mean	363.1	352.8	352.9	347.9
	SD	37.12	29.17	26.84	39.29
	Min	308.8	314.6	311.6	297.5
	Max	417.2	422.2	397.2	416.3
	N	10	10	10	10
Day 78	Mean	370.8	362.6	364.1	358.9
	SD	37.21	30.89	26.86	41.69
	Min	316.6	320.9	327.4	296.8
	Max	429.1	430.2	412.9	431.7
	N	10	10	10	10
Day 85	Mean	376.7	371.2	371.8	359.3
	SD	39.46	31.04	29.82	43.68
	Min	320.9	328.9	331.8	294.8
	Max	434.9	438.9	420.3	429.7
	N	10	10	10	10
Day 90	Mean	384.1	378.0	379.0	370.1
	SD	39.17	32.20	27.96	44.67
	Min	331.9	337.4	340.5	305.5
	Max	443.9	451.6	426.4	446.9
	N	10	10	10	10

MALES (Recovery)

RECOVERY (R)		Group 1R	Group 4R	RECOVERY (R)		Group 1R	Group 4R
Day 1	Mean	180.6	177.8	Day 36	Mean	297.2	294.6
	SD	5.01	7.87		SD	17.46	18.29
	Min	172.1	164.5		Min	254.7	256.7
	Max	189.1	189.0		Max	316.8	318.1
	N	10	10		N	10	10
Day 8	Mean	212.6	209.3	Day 43	Mean	313.7	313.3
	SD	7.89	11.72		SD	19.34	20.33
	Min	198.7	196.1		Min	273.9	272.7
	Max	224.0	226.5		Max	336.9	343.9
	N	10	10		N	10	10
Day 15	Mean	236.5	236.3	Day 50	Mean	327.8	322.5
	SD	9.46	11.46		SD	21.69	23.41
	Min	214.6	220.6		Min	284.8	275.9
	Max	249.9	257.4		Max	352.8	359.4
	N	10	10		N	10	10
Day 22	Mean	261.0	259.0	Day 57	Mean	338.3	332.5
	SD	12.75	14.34		SD	23.93	27.96
	Min	232.2	237.8		Min	294.1	285.4
	Max	280.8	280.1		Max	368.6	372.9
	N	10	10		N	10	10
Day 29	Mean	281.5	278.0	Day 64	Mean	349.8	344.5
	SD	14.56	15.18		SD	24.35	26.92
	Min	246.3	252.9		Min	304.4	296.7
	Max	301.1	296.5		Max	382.6	380.5
	N	10	10		N	10	10

RECOVERY (R)		Group 1R	Group 4R	RECOVERY (R)		Group 1R	Group 4R
Day 71	Mean	360.1	356.9	Day 98	Mean	386.7	385.5
	SD	24.83	26.86		SD	26.17	28.46
	Min	313.7	311.0		Min	338.1	329.4
	Max	396.6	391.7		Max	426.7	412.6
	N	10	10		N	10	10
Day 78	Mean	367.6	362.6	Day 105	Mean	394.6	394.5
	SD	24.78	28.60		SD	27.87	29.04
	Min	321.3	311.0		Min	342.9	337.2
	Max	402.9	401.7		Max	437.5	421.3
	N	10	10		N	10	10
Day 85	Mean	373.8	363.1	Day 112	Mean	405.0	404.9
	SD	26.40	30.75		SD	28.67	29.22
	Min	325.0	310.2		Min	352.1	349.6
	Max	413.8	409.3		Max	451.8	436.8
	N	10	10		N	10	10
Day 91	Mean	381.7	379.5	Day 118	Mean	410.9	411.2
	SD	26.10	28.33		SD	31.05	29.69
	Min	337.7	328.7		Min	355.9	356.7
	Max	425.7	410.4		Max	464.1	443.5
	N	10	10		N	10	10

BODY WEIGHTS (G)

FEMALES

TREATMENT		Group 1	Group 2	Group 3	Group 4
Day 1	Mean	142.7	143.1	142.5	141.0
	SD	5.75	4.32	6.97	4.73
	Min	132.6	136.9	133.7	135.2
	Max	153.2	149.1	158.2	150.6
	N	10	10	10	10
Day 8	Mean	160.8	160.7	158.7	155.1
	SD	8.33	6.72	6.46	6.78
	Min	147.5	148.8	146.3	146.1
	Max	172.5	170.5	170.2	168.8
	N	10	10	10	10
Day 15	Mean	171.4	172.5	170.5	168.2
	SD	7.85	6.56	6.55	6.88
	Min	159.7	165.3	159.2	154.7
	Max	184.0	185.7	181.1	181.8
	N	10	10	10	10
Day 22	Mean	182.1	183.6	182.9	178.5
	SD	7.63	9.12	7.35	6.63
	Min	168.4	173.2	171.0	166.7
	Max	193.3	201.6	194.0	189.2
	N	10	10	10	10
Day 29	Mean	191.8	191.6	191.2	184.2
	SD	9.63	11.77	8.25	6.00
	Min	177.8	178.5	180.4	176.5
	Max	202.6	219.8	203.7	195.3
	N	10	10	10	10

TREATMENT		Group 1	Group 2	Group 3	Group 4
Day 36	Mean	195.5	197.9	197.4	187.4
	SD	10.33	13.76	14.21	7.91
	Min	180.3	178.8	175.3	177.7
	Max	206.1	224.1	217.9	203.5
	N	10	10	10	10
Day 43	Mean	203.6	205.0	206.2	191.6*
	SD	8.63	15.85	13.37	8.80
	Min	185.7	184.7	183.6	181.6
	Max	212.1	236.7	225.3	208.2
	N	10	10	10	10
Day 50	Mean	211.2	211.2	212.3	199.0
	SD	9.88	16.82	15.44	9.84
	Min	193.3	189.3	192.9	187.1
	Max	221.0	240.9	238.9	217.2
	N	10	10	10	10
Day 57	Mean	213.9	215.4	214.6	200.5
	SD	9.51	16.66	16.37	11.49
	Min	195.5	191.8	193.1	186.8
	Max	225.8	246.3	244.5	218.1
	N	10	10	10	10
Day 64	Mean	216.3	217.6	216.4	203.0
	SD	8.83	16.20	17.62	11.03
	Min	202.9	197.3	196.4	188.3
	Max	230.7	246.9	252.9	221.1
	N	10	10	10	10

* Significant at p ≤ 0.05 with Group 3,

TREATMENT		Group 1	Group 2	Group 3	Group 4
Day 71	Mean	219.8	222.8	219.6	208.7
	SD	7.42	20.63	18.07	12.81
	Min	210.2	197.5	197.8	190.3
	Max	233.4	270.8	257.2	229.4
	N	10	10	10	10
Day 78	Mean	222.0	224.9	220.7	208.6
	SD	7.33	21.17	19.66	11.54
	Min	212.6	197.8	198.2	192.1
	Max	235.4	274.3	262.9	226.4
	N	10	10	10	10
Day 85	Mean	226.1	226.5	223.8	206.2*
	SD	8.76	21.09	19.26	13.37
	Min	214.5	199.3	202.9	194.3
	Max	242.0	275.4	266.1	229.3
	N	10	10	10	10
Day 90	Mean	228.3	228.3	226.2	212.8
	SD	8.70	21.19	19.34	11.67
	Min	215.3	200.2	203.8	199.1
	Max	242.3	276.6	267.4	231.0
	N	10	10	10	10

Significant at p ≤ 0.05 with Groups 1, 2 and 3

BODY WEIGHTS (G) (Recovery)

FEMALES

RECOVERY (R)		Group 1R	Group 4R	RECOVERY (R)		Group 1R	Group 4R
Day 1	Mean	141.1	141.8	Day 36	Mean	194.5	193.2
	SD	4.46	5.58		SD	10.05	10.45
	Min	133.2	133.3		Min	182.3	174.7
	Max	146.2	150.8		Max	210.2	212.0
	N	10	10		N	10	10
Day 8	Mean	159.4	156.9	Day 43	Mean	202.7	199.9
	SD	8.07	6.82		SD	11.53	8.43
	Min	146.4	144.3		Min	184.1	188.8
	Max	168.7	167.7		Max	221.3	215.7
	N	10	10		N	10	10
Day 15	Mean	172.4	167.5	Day 50	Mean	211.6	206.0
	SD	8.21	5.53		SD	7.90	10.94
	Min	164.1	156.0		Min	197.6	189.4
	Max	191.2	175.2		Max	223.1	224.8
	N	10	10		N	10	10
Day 22	Mean	182.7	181.4	Day 57	Mean	214.1	209.2
	SD	8.71	6.65		SD	9.64	14.64
	Min	174.6	169.3		Min	198.3	188.4
	Max	199.5	190.5		Max	228.4	238.2
	N	10	10		N	10	10
Day 29	Mean	190.6	187.0	Day 64	Mean	215.6	210.2
	SD	10.05	7.01		SD	8.47	15.02
	Min	175.1	174.7		Min	201.6	189.2
	Max	207.3	197.1		Max	229.0	238.4
	N	10	10		N	10	10

RECOVERY (R)		Group 1R	Group 4R	RECOVERY (R)		Group 1R	Group 4R
Day 71	Mean	218.0	214.6	Day 98	Mean	228.5	219.5
	SD	8.04	17.24		SD	11.30	16.21
	Min	202.5	196.2		Min	207.6	200.7
	Max	230.0	249.9		Max	250.1	252.1
	N	10	10		N	10	10
Day 78	Mean	220.8	213.2	Day 105	Mean	231.3	221.2
	SD	9.31	15.77		SD	11.36	17.27
	Min	202.9	192.6		Min	209.6	201.3
	Max	234.8	246.1		Max	251.2	257.2
	N	10	10		N	10	10
Day 85	Mean	221.4	210.6	Day 112	Mean	234.2	223.0
	SD	10.34	18.01		SD	12.05	17.97
	Min	204.3	188.6		Min	210.2	202.9
	Max	235.1	248.1		Max	252.6	260.2
	N	10	10		N	10	10
Day 91	Mean	224.6	217.3	Day 118	Mean	236.4	225.5
	SD	13.15	16.76		SD	12.30	17.82
	Min	206.4	198.7		Min	211.7	205.2
	Max	249.4	251.9		Max	255.2	261.9
	N	10	10		N	10	10

BODY WEIGHT GAIN (%)

MALES

TREATMENT		Group 1	Group 2	Group 3	Group 4
Day 1	Mean	0.0	0.0	0.0	0.0
	SD	0.00	0.00	0.00	0.00
	Min	0.0	0.0	0.0	0.0
	Max	0.0	0.0	0.0	0.0
	N	10	10	10	10
Day 8	Mean	18.6	15.4	19.0	19.8
	SD	3.08	5.29	2.76	4.70
	Min	12.4	6.5	16.5	14.4
	Max	22.4	22.8	25.7	27.1
	N	10	10	10	10
Day 15	Mean	34.8	30.9	31.6	32.1
	SD	5.64	4.27	5.90	9.10
	Min	26.4	24.7	22.9	22.5
	Max	44.2	39.7	43.4	47.7
	N	10	10	10	10
Day 22	Mean	48.4	42.0	42.3	41.6
	SD	6.68	4.78	7.20	10.88
	Min	40.4	35.8	28.4	29.9
	Max	61.3	50.2	55.3	61.7
	N	10	10	10	10
Day 29	Mean	62.0	52.4	52.0	52.8
	SD	7.28	6.26	7.76	12.62
	Min	53.5	42.2	34.7	39.3
	Max	74.0	65.6	61.3	77.4
	N	10	10	10	10

TREATMENT		Group 1	Group 2	Group 3	Group 4
Day 36	Mean	72.3	59.9	59.7	63.7
	SD	11.78	10.06	9.13	14.24
	Min	56.3	45.6	42.6	47.3
	Max	93.3	83.8	71.7	91.0
	N	10	10	10	10
Day 43	Mean	81.3	69.9	67.7	69.3
	SD	13.13	10.62	11.13	16.52
	Min	58.5	56.6	48.2	48.3
	Max	96.3	94.7	83.6	99.9
	N	10	10	10	10
Day 50	Mean	86.6	76.4	74.2	76.7
	SD	14.82	11.27	12.26	19.31
	Min	61.4	61.5	52.2	51.9
	Max	103.3	102.2	93.1	111.4
	N	10	10	10	10
Day 57	Mean	92.7	82.7	80.2	81.7
	SD	17.52	13.33	14.11	18.73
	Min	62.0	67.8	58.0	58.2
	Max	112.9	114.3	101.8	113.1
	N	10	10	10	10
Day 64	Mean	98.3	89.3	86.9	88.0
	SD	17.87	14.99	14.32	19.85
	Min	67.1	71.0	65.9	61.8
	Max	118.2	122.4	110.7	121.0
	N	10	10	10	10

TREATMENT		Group 1	Group 2	Group 3	Group 4
Day 71	Mean	103.9	94.5	93.7	94.2
	SD	19.06	16.31	16.01	20.06
	Min	70.2	75.7	68.3	66.4
	Max	123.6	132.1	119.2	130.3
	N	10	10	10	10
Day 78	Mean	108.2	99.9	99.9	100.4
	SD	19.20	17.35	16.61	21.33
	Min	74.5	79.2	76.9	64.4
	Max	129.4	136.5	127.9	138.8
	N	10	10	10	10
Day 85	Mean	111.6	104.7	104.1	100.5
	SD	20.41	17.46	17.95	21.84
	Min	76.9	83.6	79.3	70.0
	Max	132.8	141.3	132.0	137.7
	N	10	10	10	10
Day 90	Mean	115.7	108.4	108.1	106.6
	SD	19.96	17.97	17.61	22.45
	Min	83.0	88.4	84.0	74.2
	Max	137.0	148.3	135.3	147.2
	N	10	10	10	10

MALES

RECOVERY (R)		Group 1R	Group 4R	RECOVERY (R)		Group 1R	Group 4R
Day 1	Mean	0.0	0.0	Day 36	Mean	64.5	65.8
	SD	0.00	0.00		SD	8.55	9.37
	Min	0.0	0.0		Min	48.0	55.4
	Max	0.0	0.0		Max	76.3	80.2
	N	10	10		N	10	10
Day 8	Mean	17.7	17.8	Day 43	Mean	73.7	76.5
	SD	3.82	5.54		SD	10.36	12.11
	Min	12.5	6.9		Min	59.2	64.2
	Max	23.5	27.4		Max	91.3	94.8
	N	10	10		N	10	10
Day 15	Mean	30.9	33.1	Day 50	Mean	81.5	81.6
	SD	3.83	6.58		SD	11.37	13.43
	Min	23.9	20.3		Min	64.9	67.0
	Max	36.4	44.8		Max	100.3	99.8
	N	10	10		N	10	10
Day 22	Mean	44.4	45.7	Day 57	Mean	87.3	87.3
	SD	4.83	6.59		SD	12.86	16.23
	Min	34.9	31.5		Min	67.8	64.0
	Max	49.8	57.5		Max	109.3	107.7
	N	10	10		N	10	10
Day 29	Mean	55.8	56.5	Day 64	Mean	93.7	94.0
	SD	6.49	8.27		SD	13.22	15.06
	Min	43.1	37.9		Min	73.6	69.6
	Max	65.0	68.0		Max	117.3	113.6
	N	10	10		N	10	10

RECOVERY (R)		Group 1R	Group 4R	RECOVERY (R)		Group 1R	Group 4R
Day 71	Mean	99.4	100.9	Day 98	Mean	114.2	117.0
	SD	13.77	14.49		SD	14.66	14.59
	Min	79.5	76.6		Min	94.0	92.7
	Max	125.2	117.7		Max	142.3	133.8
	N	10	10		N	10	10
Day 78	Mean	103.5	104.0	Day 105	Mean	118.5	122.0
	SD	13.55	14.14		SD	15.76	14.94
	Min	84.4	79.8		Min	97.5	95.8
	Max	128.8	123.3		Max	148.4	138.5
	N	10	10		N	10	10
Day 85	Mean	107.0	104.3	Day 112	Mean	124.3	127.9
	SD	14.42	14.84		SD	16.39	15.54
	Min	86.7	77.5		Min	103.5	99.6
	Max	135.0	127.5		Max	156.6	148.1
	N	10	10		N	10	10
Day 91	Mean	111.4	113.6	Day 118	Mean	127.6	131.5
	SD	14.93	14.69		SD	18.05	16.06
	Min	90.5	86.1		Min	106.0	102.8
	Max	141.7	129.6		Max	163.5	152.3
	N	10	10		N	10	10

BODY WEIGHTS GAIN (%)

FEMALES

TREATMENT		Group 1	Group 2	Group 3	Group 4
Day 1	Mean	0.0	0.0	0.0	0.0
	SD	0.00	0.00	0.00	0.00
	Min	0.0	0.0	0.0	0.0
	Max	0.0	0.0	0.0	0.0
	N	10	10	10	10
Day 8	Mean	12.7	12.3	11.5	10.0
	SD	4.58	4.60	5.14	5.05
	Min	6.0	3.8	4.4	1.1
	Max	20.9	16.9	18.7	15.8
	N	10	10	10	10
Day 15	Mean	20.2	20.6	19.8	19.3
	SD	4.76	5.52	5.55	4.84
	Min	11.3	10.9	10.9	9.3
	Max	28.1	31.1	29.1	25.1
	N	10	10	10	10
Day 22	Mean	27.7	28.4	28.5	26.7
	SD	4.60	7.27	5.96	5.27
	Min	17.3	16.2	15.9	17.8
	Max	34.6	42.4	34.7	32.6
	N	10	10	10	10
Day 29	Mean	34.5	34.0	34.3	30.7
	SD	5.11	9.01	5.72	4.10
	Min	23.8	21.8	22.2	24.7
	Max	40.8	55.2	40.4	36.3
	N	10	10	10	10

TREATMENT		Group 1	Group 2	Group 3	Group 4
Day 36	Mean	37.1	38.4	38.7	32.9
	SD	6.27	10.38	9.64	3.32
	Min	28.6	22.0	18.8	25.6
	Max	46.7	58.3	52.1	37.6
	N	10	10	10	10
Day 43	Mean	42.9	43.4	44.9	35.9
	SD	7.20	12.64	9.53	4.95
	Min	32.8	26.0	29.5	28.3
	Max	52.9	67.2	58.1	44.1
	N	10	10	10	10
Day 50	Mean	48.2	47.7	49.1	41.1
	SD	7.05	12.69	10.02	4.21
	Min	34.6	29.1	32.4	35.5
	Max	57.0	70.1	67.6	46.9
	N	10	10	10	10
Day 57	Mean	50.0	50.7	50.8	42.2
	SD	7.07	12.84	10.98	5.36
	Min	36.1	30.8	32.9	33.4
	Max	60.7	73.9	71.6	49.1
	N	10	10	10	10
Day 64	Mean	51.7	52.1	52.0	43.9
	SD	7.01	11.95	11.85	5.66
	Min	41.7	34.6	33.8	35.8
	Max	64.2	74.4	77.5	51.2
	N	10	10	10	10

TREATMENT		Group 1	Group 2	Group 3	Group 4
Day 71	Mean	54.2	55.8	54.3	48.0
	SD	6.86	15.72	12.36	6.46
	Min	43.4	34.7	36.9	37.2
	Max	66.1	91.2	80.5	58.5
	N	10	10	10	10
Day 78	Mean	55.8	57.3	55.0	47.9
	SD	7.07	15.83	13.40	5.50
	Min	44.3	34.9	37.9	38.5
	Max	67.5	93.7	84.5	55.8
	N	10	10	10	10
Day 85	Mean	58.6	58.4	57.3	46.1*
	SD	7.56	15.95	13.59	6.22
	Min	48.0	35.9	37.7	39.9
	Max	72.2	94.5	86.7	58.5
	N	10	10	10	10
Day 90	Mean	60.2	59.7	59.0	50.9
	SD	7.56	16.06	14.14	5.52
	Min	49.8	36.6	38.1	43.5
	Max	72.5	95.3	87.6	61.0
	N	10	10	10	10

* Significant at p ≤ 0.05 with Groups 1, 2 and 3

FEMALES (Recovery)

RECOVERY (R)		Group 1R	Group 4R	RECOVERY (R)		Group 1R	Group 4R
Day 1	Mean	0.0	0.0	Day 36	Mean	37.9	36.3
	SD	0.00	0.00		SD	8.00	6.49
	Min	0.0	0.0		Min	25.2	29.2
	Max	0.0	0.0		Max	49.8	47.6
	N	10	10		N	10	10
Day 8	Mean	13.0	10.6	Day 43	Mean	43.8	41.1
	SD	4.65	1.66		SD	9.95	6.82
	Min	4.8	8.3		Min	25.9	31.1
	Max	18.7	13.5		Max	57.7	50.2
	N	10	10		N	10	10
Day 15	Mean	22.2	18.2	Day 50	Mean	50.1	45.4
	SD	6.41	2.16		SD	7.03	6.97
	Min	15.8	13.9		Min	37.7	37.5
	Max	37.2	21.4		Max	59.0	56.5
	N	10	10		N	10	10
Day 22	Mean	29.5	28.0	Day 57	Mean	51.8	47.5
	SD	7.46	2.83		SD	7.59	8.60
	Min	20.2	23.5		Min	41.2	39.9
	Max	43.1	31.3		Max	62.8	65.9
	N	10	10		N	10	10
Day 29	Mean	35.2	31.9	Day 64	Mean	52.9	48.2
	SD	8.03	4.04		SD	7.00	8.28
	Min	27.5	25.5		Min	41.7	37.4
	Max	47.8	37.3		Max	63.2	66.0
	N	10	10		N	10	10

RECOVERY (R)		Group 1R	Group 4R	RECOVERY (R)		Group 1R	Group 4R
Day 71	Mean	54.6	51.3	Day 98	Mean	62.1	54.8
	SD	7.19	10.08		SD	10.74	9.16
	Min	43.1	38.3		Min	42.0	43.9
	Max	65.2	74.0		Max	78.3	75.6
	N	10	10		N	10	10
Day 78	Mean	56.6	50.3	Day 105	Mean	64.1	56.0
	SD	8.08	9.37		SD	11.09	9.85
	Min	43.8	40.4		Min	43.4	44.1
	Max	68.7	71.4		Max	79.6	79.1
	N	10	10		N	10	10
Day 85	Mean	57.1	48.5	Day 112	Mean	66.2	57.2
	SD	9.38	10.47		SD	11.67	10.20
	Min	40.8	38.5		Min	43.8	44.5
	Max	71.3	72.8		Max	84.8	81.2
	N	10	10		N	10	10
Day 91	Mean	59.3	53.2	Day 118	Mean	67.7	59.0
	SD	11.19	9.64		SD	11.87	10.04
	Min	41.2	41.4		Min	44.8	47.2
	Max	77.8	75.4		Max	86.5	82.4
	N	10	10		N	10	10

FUNCTIONAL OBSERVATION BATTERY

MALES

TREATMENT(WEEK -13)		Grip Strength	Rotarod	IR Activity
TREATMENT(WEEK -13)		Fore limb	In seconds	S - Movement	F - Movement	S - Steriotypic	F - Steriotypic
Group 1	Mean	469.1	63	54.0	133.0	22.9	111.9
Group 1	SD	31.60	4.0	9.83	38.59	10.77	29.45
Group 2	Mean	462.3	63	53.4	71.0	21.7	89.7
Group 2	SD	96.44	4.5	13.36	45.30	1.67	30.15
Group 3	Mean	442.0	62	65.1	105.8	20.3	102.2
Group 3	SD	57.44	5.7	9.98	56.93	2.52	29.89
Group 4	Mean	478.1	65	56.9	119.7	20.7	103.2
Group 4	SD	80.85	4.0	11.39	71.62	2.04	29.75
Group 1R	Mean	585.9	58	54.9	161.3	38.7	106.8
Group 1R	SD	42.19	9.72	11.20	55.05	23.26	33.45
Group 4R	Mean	602.1	64	80.6	180.4	39.2	124.8
Group 4R	SD	62.81	2.96	35.20	63.33	29.06	26.77

FEMALES

TREATMENT(WEEK -13)		Grip Strength	Rotarod	IR Activity
TREATMENT(WEEK -13)		Fore limb	In seconds	S - Movement	F - Movement	S - Steriotypic	F - Steriotypic
Group 1	Mean	506.3	65	53.2	145.3	20.8	122.3
Group 1	SD	99.43	4.8	10.53	42.91	2.62	23.81
Group 2	Mean	509.6	63	61.5	186.2	21.1	133.1
Group 2	SD	89.24	7.3	9.79	42.69	3.16	16.45
Group 3	Mean	460.8	63	58.4	177.4	20.3	119.9
Group 3	SD	40.00	3.0	10.70	108.63	3.82	40.53
Group 4	Mean	460.3	65	63.1	193.5	21.7	136.2
Group 4	SD	90.85	5.0	8.54	30.51	2.91	19.65
Group 1R	Mean	612.8	62	49.1	180.7	41.9	114.2
Group 1R	SD	50.12	8.44	10.99	40.03	25.88	30.67
Group 4R	Mean	593.9	63	54.5	171.4	42.4	118.0
Group 4R	SD	28.09	6.64	14.53	76.45	33.41	35.66

MALES (Recovery)

TREATMENT(WEEK -17)		Grip Strength	Rotarod	IR Activity
TREATMENT(WEEK -17)		Fore limb	In seconds	S - Movement	F - Movement	S - Steriotypic	F - Steriotypic
Group 1R	Mean	604.1	59	47.4	99.3	50.4	88.7
Group 1R	SD	55.03	6.5	14.73	36.07	27.06	27.13
Group 4R	Mean	630.6	64	85.7	109.1	64.5	94.4
Group 4R	SD	43.63	4.6	18.80	12.96	26.75	17.87

FEMALES (Recovery)

TREATMENT(WEEK -17)		Grip Strength	Rotarod	IR Activity
TREATMENT(WEEK -17)		Fore limb	In seconds	S - Movement	F - Movement	S - Steriotypic	F - Steriotypic
Group 1R	Mean	597.6	63	58.5	134.4	21.2	120.5
Group 1R	SD	50.64	7.0	8.96	48.79	3.12	27.58
Group 4R	Mean	602.8	64	56.2	134.5	23.4	105.9
Group 4R	SD	64.14	5.8	9.03	66.72	4.77	23.82

HEMATOLOGY

MALES

TREATMENT(WEEK -13)		RBC	Hb	PCV	MCV	MCH	MCHC
TREATMENT(WEEK -13)		x10⁶Cells/µL	g/dL	%	fL	pg	g/dL
Group 1	Mean	8.67	15.93	48.12	55.62	18.44	33.14
Group 1	SD	0.596	0.638	2.312	1.936	0.847	0.806
Group 2	Mean	8.60	15.48	47.56	55.28	18.02	32.59
Group 2	SD	0.292	0.516	1.506	0.675	0.551	0.764
Group 3	Mean	8.73	15.90	48.23	55.26	18.21	32.95
Group 3	SD	0.493	0.899	2.733	1.034	0.614	0.789
Group 4	Mean	8.56	15.62	47.10	55.04	18.26	33.15
Group 4	SD	0.397	0.509	2.085	1.004	0.576	0.737

TREATMENT(WEEK -13)		PLT	WBC	RET	DC (%)					PT	APTT
TREATMENT(WEEK -13)		x10³cells/µL	x10³cells/ µL	%	NEUT	LYMPH	MONO	EOS	BASO	Sec	Sec
Group 1	Mean	718.20	5.66	2.07	17.29	77.93^$	1.86	1.42	0.36	15.76	12.16
Group 1	SD	74.730	1.622	0.285	4.176	4.898	0.643	0.447	0.107	1.009	1.709
Group 2	Mean	706.50	6.01	1.91	21.30	73.65	2.77^*	0.83^*	0.30	15.67	12.81
Group 2	SD	86.314	1.839	0.259	6.391	6.094	0.538	0.320	0.067	0.990	1.717
Group 3	Mean	624.10	6.94	1.84	29.37^*#	65.03^*#	2.95^*	1.13	0.32	14.71	11.89
Group 3	SD	181.323	1.527	0.273	6.484	6.872	0.610	0.633	0.092	0.940	1.225
Group 4	Mean	770.80^&	5.77	2.17	26.31^*	68.22^*	2.78^*	1.29	0.26	15.26	12.78
Group 4	SD	87.519	1.322	0.321	6.552	6.697	0.786	0.522	0.084	0.650	1.495

*Significant at p ≤ 0.05 with Group 1, #- Significant at p ≤ 0.05 with Group 2^, &-Significant at p ≤ 0.05 with Group 3

MALES

RECOVERY (WEEK 17)		RBC	Hb	PCV	MCV	MCH	MCHC
RECOVERY (WEEK 17)		x10⁶Cells/µL	g/dL	%	fL	pg	g/dL
Group 1R	Mean	8.31	14.77	44.85	54.45	17.81	32.95
Group 1R	SD	0.568	0.556	2.227	1.258	0.759	0.664
Group 4R	Mean	8.63	15.19	47.38*	54.25	17.63	32.48
Group 4R	SD	0.289	0.341	1.056	0.755	0.414	0.339

RECOVERY (WEEK 17)		PLT	WBC	RET	DC (%)					PT	APTT
RECOVERY (WEEK 17)		x10³cells/µL	x10³cells/ µL	%	NEUT	LYMPH	MONO	EOS	BASO	Sec	Sec
Group 1R	Mean	808.40	4.55	2.13	26.58	68.11	2.69	1.08	0.23	15.72	12.08
Group 1R	SD	96.212	1.138	0.299	7.319	7.226	0.805	0.457	0.309	1.165	1.067
Group 4R	Mean	811.80	4.39	2.04	23.96	70.26	2.93	1.63	0.19	15.93	12.72
Group 4R	SD	77.334	1.193	0.205	6.263	6.861	0.558	0.862	0.099	1.012	1.173

^*Significant at p ≤ 0.05 with Group 1R

FEMALES

TREATMENT(WEEK -13)		RBC	Hb	PCV	MCV	MCH	MCHC
TREATMENT(WEEK -13)		x10⁶Cells/µL	g/dL	%	fL	pg	g/dL
Group 1	Mean	8.08	14.92	45.80	56.76	18.47	32.58
Group 1	SD	0.468	0.464	2.076	1.213	0.633	0.705
Group 2	Mean	7.90	14.69	44.62	56.49	18.57	32.88
Group 2	SD	0.234	0.493	1.352	0.656	0.467	0.620
Group 3	Mean	8.00	15.09	45.67	57.10	18.88	33.05
Group 3	SD	0.295	0.314	1.432	1.140	0.663	0.776
Group 4	Mean	7.90	15.02	45.34	57.43	19.02	33.12
Group 4	SD	0.279	0.421	1.490	1.059	0.555	0.668

TREATMENT(WEEK -13)		PLT	WBC	RET	DC (%)					PT	APTT
TREATMENT(WEEK -13)		x10³cells/µL	x10³cells/ µL	%	NEUT	LYMPH	MONO	EOS	BASO	Sec	Sec
Group 1	Mean	823.80	3.39	1.97	15.77	79.65	2.05	1.16	0.27	15.70	12.74
Group 1	SD	177.020	0.615	0.398	4.606	4.918	0.357	0.237	0.125	0.738	1.695
Group 2	Mean	874.80	3.80	2.25	16.73	78.68	2.15	0.98	0.28	15.17	12.42
Group 2	SD	123.591	0.973	0.368	3.823	4.102	0.562	0.249	0.140	0.962	1.040
Group 3	Mean	834.90	3.39	2.13	19.29	76.10	2.28	0.96	0.25	15.43	12.83
Group 3	SD	66.542	0.684	0.350	6.391	6.784	0.625	0.227	0.085	0.538	1.491
Group 4	Mean	804.20	3.46	2.30	23.41*^#	71.28*^#	2.61	1.25	0.26	15.15	13.03
Group 4	SD	78.816	1.787	0.530	5.895	5.780	0.465	0.631	0.324	0.682	1.411

^*Significant at p ≤ 0.05 with Group 1, #-Significant at p ≤ 0.05 with Group 2

FEMALES

RECOVERY (WEEK 17)		RBC	Hb	PCV	MCV	MCH	MCHC
RECOVERY (WEEK 17)		x10⁶Cells/µL	g/dL	%	fL	pg	g/dL
Group 1R	Mean	7.96	14.81	45.02	56.55	18.57	32.88
Group 1R	SD	0.290	0.463	1.521	0.848	0.490	0.598
Group 4R	Mean	8.15	14.99	46.65	56.63	18.20	32.52
Group 4R	SD	0.476	0.456	1.434	1.821	0.876	0.611

RECOVERY (WEEK 17)		PLT	WBC	RET	DC (%)					PT	APTT
RECOVERY (WEEK 17)		x10³cells/µL	x10³cells/ µL	%	NEUT	LYMPH	MONO	EOS	BASO	Sec	Sec
Group 1R	Mean	833.10	2.78	1.78	13.63	81.82	1.99	1.37	0.14	16.32	13.11
Group 1R	SD	147.485	0.693	0.220	1.310	7.344	0.191	0.744	0.097	0.665	0.971
Group 4R	Mean	758.30	1.68*	1.64	18.22	76.59	2.33	1.73	0.14	15.92	12.93
Group 4R	SD	96.524	0.441	0.179	4.682	5.053	0.576	0.538	0.117	0.866	1.108

*Significant at p ≤ 0.05 with Group 1R

CLINICAL BIOCHEMISTRY (CONTD.)

MALES

TREATMENT(WEEK 13)		GLU	UREA	BUN	CREA	CHOL	TRIGL	AST	ALT	ALP
TREATMENT(WEEK 13)		mmol/L	mmol/L	mmol/L	µmol/L	mmol/L	mmol/L	U/L	U/L	U/L
Group 1	Mean	9.51	5.48	2.56	43.06	1.81	0.53	62.00	34.13	77.42
Group 1	SD	1.754	1.219	0.570	8.132	0.350	0.190	5.219	4.551	26.735
Group 2	Mean	10.21	5.53	2.59	37.86	1.97	0.69	61.70	34.80	88.55
Group 2	SD	2.161	1.512	0.706	5.190	0.284	0.245	5.582	4.177	21.158
Group 3	Mean	9.76	6.01	2.81	47.12^#	2.02	1.03	75.40*^#	41.87	133.35^*
Group 3	SD	1.813	1.199	0.560	6.666	0.460	1.096	13.237	9.319	66.920
Group 4	Mean	9.11	5.76	2.67	46.12	2.00	0.70	81.76*^#	45.30^*	122.34^*
Group 4	SD	1.129	0.899	0.401	9.777	0.407	0.263	14.515	13.528	59.983

TREATMENT(WEEK 13)		BIL	GGT	Na	K	Cl	TPO	ALB	GLB	A/G
TREATMENT(WEEK 13)		µmol/L	U/L	mmol/L	mmol/L	mmol/L	g/L	g/L	g/L	_
Group 1	Mean	1.48	0.00	142.26	3.64	108.03	67.57	44.74	22.84	2.04
Group 1	SD	0.710	0.00	1.095	0.141	0.762	3.973	2.785	3.824	0.554
Group 2	Mean	0.85	0.00	141.21	3.70	107.94	66.82	43.73	23.09	1.90
Group 2	SD	0.620	0.00	1.526	0.259	0.991	3.612	3.176	1.287	0.158
Group 3	Mean	1.03	0.00	142.07	3.79	107.87	67.63	43.83	24.32	1.86
Group 3	SD	0.773	0.00	1.470	0.348	0.913	3.246	1.984	3.275	0.276
Group 4	Mean	1.18	0.00	141.89	3.84	108.28	66.10	44.20	21.90	2.03
Group 4	SD	0.719	0.00	1.265	0.429	0.399	2.692	1.213	1.733	0.125

*-Significant at p ≤ 0.05 with Group 1, # - Significant at p ≤ 0.05 with Group 2.

MALES

RECOVERY (WEEK 17)		GLU	UREA	BUN	CREA	CHOL	TRIGL	AST	ALT	ALP
RECOVERY (WEEK 17)		mmol/L	mmol/L	mmol/L	µmol/L	mmol/L	mmol/L	U/L	U/L	U/L
Group 1R	Mean	10.54	6.33	2.96	41.13	2.10	0.62	60.67	34.29	65.86
Group 1R	SD	1.389	0.800	0.374	4.603	0.318	0.125	14.096	15.181	9.311
Group 4R	Mean	9.76	6.28	2.94	38.20	2.27	0.72	61.80	32.32	53.69*
Group 4R	SD	1.132	1.147	0.536	4.274	0.171	0.174	9.370	5.683	11.373

RECOVERY (WEEK 17)		BIL	GGT	Na	K	Cl	TPO	ALB	GLB	A/G
RECOVERY (WEEK 17)		µmol/L	U/L	mmol/L	mmol/L	mmol/L	g/L	g/L	g/L	_
Group 1R	Mean	1.03	0.00	145.12	3.85	106.84	63.74	41.47	22.27	2.22
Group 1R	SD	0.589	0.00	1.137	0.408	1.206	1.817	5.333	5.806	1.613
Group 4R	Mean	0.76	0.00	145.47	3.89	106.92	64.70	43.52	21.18	2.06
Group 4R	SD	0.540	0.00	0.943	0.300	0.884	1.791	2.282	1.128	0.198

*-Significant at p ≤ 0.05 with Group 1R

FEMALES

TREATMENT(WEEK 13)		GLU	UREA	BUN	CREA	CHOL	TRIGL	AST	ALT	ALP
TREATMENT(WEEK 13)		mmol/L	mmol/L	mmol/L	µmol/L	mmol/L	mmol/L	U/L	U/L	U/L
Group 1	Mean	7.66	6.28	2.94	43.85	1.62	0.48	60.84	26.78	40.67
Group 1	SD	1.816	2.645	1.236	11.498	0.387	0.176	9.058	6.606	29.926
Group 2	Mean	9.18	6.51	3.04	45.10	1.77	0.51	61.47	29.19	39.15
Group 2	SD	1.686	1.475	0.689	5.859	0.358	0.089	7.767	5.150	14.333
Group 3	Mean	8.73	6.89	3.22	50.84^*	1.86	0.59	79.24*^#	29.03	57.04
Group 3	SD	1.079	1.168	0.546	7.642	0.433	0.193	16.562	8.981	40.442
Group 4	Mean	9.39	7.37	3.51	47.81	1.70	0.62	69.84	30.18	44.01
Group 4	SD	1.404	1.354	0.637	6.199	0.317	0.365	10.845	10.816	22.765

TREATMENT(WEEK 13)		BIL	GGT	Na	K	Cl	TPO	ALB	GLB	A/G
TREATMENT(WEEK 13)		µmol/L	U/L	mmol/L	mmol/L	mmol/L	g/L	g/L	g/L	_
Group 1	Mean	1.64	0.00	140.46	3.44	107.85	69.48	50.21	19.27	2.64
Group 1	SD	0.990	0.00	1.153	0.163	1.801	4.462	4.020	2.315	0.390
Group 2	Mean	0.96	0.00	140.20	3.48	107.42	71.99	51.58	20.42	2.62
Group 2	SD	0.398	0.00	1.391	0.246	1.900	3.685	4.016	3.709	0.573
Group 3	Mean	1.39	0.00	140.97	3.49	107.76	71.60	50.55	21.05	2.45
Group 3	SD	0.930	0.00	1.018	0.529	0.706	3.045	3.468	2.581	0.431
Group 4	Mean	1.38	0.00	140.05	3.58	108.20	70.38	51.71	18.68	2.79
Group 4	SD	0.972	0.00	1.400	0.520	1.194	4.139	4.585	1.509	0.408

^*- Significant at p ≤ 0.05 with Group 1,^#-Significant at p ≤ 0.05 with Group 2.

FEMALES

RECOVERY (WEEK 17)		GLU	UREA	BUN	CREA	CHOL	TRIGL	AST	ALT	ALP
RECOVERY (WEEK 17)		mmol/L	mmol/L	mmol/L	µmol/L	mmol/L	mmol/L	U/L	U/L	U/L
Group 1R	Mean	9.58	8.12	3.80	40.86	1.53	0.56	65.66	31.59	25.98
Group 1R	SD	1.565	0.903	0.368	2.538	0.378	0.074	23.576	10.057	10.694
Group 4R	Mean	9.09	9.33*	4.36*	45.81*	2.18	0.47	61.93	26.50	23.38
Group 4R	SD	1.623	0.926	0.433	4.525	0.355	0.115	11.343	5.049	5.666

RECOVERY (WEEK 17)		BIL	GGT	Na	K	Cl	TPO	ALB	GLB	A/G
RECOVERY (WEEK 17)		µmol/L	U/L	mmol/L	mmol/L	mmol/L	g/L	g/L	g/L	_
Group 1R	Mean	1.31	0.00	144.31	3.49	106.33	68.42	49.61	18.81	2.68
Group 1R	SD	0.854	0.00	1.318	0.199	1.193	3.014	3.541	1.981	0.462
Group 4R	Mean	1.00	0.00	144.34	3.50	106.42	72.16	53.58*	18.58	3.26
Group 4R	SD	0.566	0.00	1.582	0.299	2.126	7.046	3.022	4.991	1.698

^*Significant at p ≤ 0.05 with Group 1R.

URINE ANALYSIS

MALES

TREATMENT (Week 13)		Volume	SG	pH	Pro	GLU	KET	UBG	BIL	Color	Clarity
TREATMENT (Week 13)		mL	-	-	mg/dl	-	mg/dl	mg/dl	mg/dl	-	-
Group 1	Mean	5.97	1.02	8.50	37.50	0.00	5.00	0.00	0.00	NA	NA
Group 1	SD	0.691	0.002	0.527	39.528	0.000	4.082	0.000	0.000	NA	NA
Group 2	Mean	5.54	1.01*	8.20	17.50	0.00	3.00	0.00	0.00	NA	NA
Group 2	SD	0.629	0.003	1.033	12.076	0.000	2.582	0.000	0.000	NA	NA
Group 3	Mean	5.43	1.02*	8.15	17.50	0.00	3.50	0.00	0.10	NA	NA
Group 3	SD	0.519	0.005	1.375	12.076	0.000	4.743	0.000	0.316	NA	NA
Group 4	Mean	5.10*	1.02	8.35	50.00	0.00	4.00	0.00	0.00	NA	NA
Group 4	SD	0.556	0.003	0.944	44.096	0.000	4.595	0.000	0.000	NA	NA

TREATMENT (WEEK 13)		ERY	LEU	EPI	RBC	PUS	CAST	CRY	SPER
TREATMENT (WEEK 13)		/µL	/µL	/field	/field	/field	/field	/field	/field
Group 1	Mean	0.00	117.50	0.90	0.00	0.00	0.00	2.30	0.00
Group 1	SD	0.000	138.969	0.316	0.000	0.000	0.000	0.823	0.000
Group 2	Mean	0.00	125.00	0.90	0.00	0.00	0.00	2.50	0.00
Group 2	SD	0.000	135.401	0.316	0.000	0.000	0.000	0.850	0.000
Group 3	Mean	1.00	67.50	1.00	0.00	0.00	0.00	2.20	0.00
Group 3	SD	3.162	42.573	0.000	0.000	0.000	0.000	1.033	0.000
Group 4	Mean	1.00	150.00	1.00	0.00	0.00	0.00	2.10	0.00
Group 4	SD	3.162	187.824	0.000	0.000	0.000	0.000	0.738	0.000

Key: NA = Not Applicable,*Significant at p ≤ 0.05 with Group 1.

MALES

RECOVERY (Week 17)		Volume	SG	pH	Pro	GLU	KET	UBG	BIL	Color	Clarity
RECOVERY (Week 17)		mL	-	-	mg/dl	-	mg/dl	mg/dl	mg/dl	-	-
Group 1R	Mean	5.82	1.02	8.20	70.00	0.00	4.50	0.10	0.00	NA	NA
Group 1R	SD	0.469	0.003	0.789	151.291	0.000	4.378	0.316	0.000	NA	NA
Group 4R	Mean	5.41	1.01	8.40	42.50	0.00	4.50	0.00	0.00	NA	NA
Group 4R	SD	0.769	0.002	0.699	28.988	0.000	4.378	0.000	0.000	NA	NA

RECOVERY (Week 17)		ERY	LEU	EPI	RBC	PUS	CAST	CRY	SPER
RECOVERY (Week 17)		/µL	/µL	/field	/field	/field	/field	/field	/field
Group 1R	Mean	12.50	80.00	0.90	0.00	0.00	0.00	2.60	0.00
Group 1R	SD	17.678	149.443	0.316	0.000	0.000	0.000	0.843	0.000
Group 4R	Mean	1.00	117.50	1.00	0.00	0.00	0.00	2.70	0.20
Group 4R	SD	3.162	138.969	0.000	0.000	0.000	0.000	0.823	0.422

Key: NA = Not Applicable

FEMALES

TREATMENT (Week 13)		Volume	SG	pH	Pro	GLU	KET	UBG	BIL	Color	Clarity
TREATMENT (Week 13)		mL	-	-	mg/dl	-	mg/dl	mg/dl	mg/dl	-	-
Group 1	Mean	5.36	1.02	8.60	85.00	0.00	2.50	0.10	0.00	NA	NA
Group 1	SD	0.602	0.000	0.516	153.749	0.000	4.859	0.316	0.000	NA	NA
Group 2	Mean	5.82	1.02	8.80	37.50	0.00	2.00	0.00	0.00	NA	NA
Group 2	SD	0.643	0.003	0.422	44.49	0.000	4.830	0.000	0.000	NA	NA
Group 3	Mean	5.41	1.02	8.90	20.00	0.00	3.50	0.00	0.00	NA	NA
Group 3	SD	0.540	0.003	0.316	22.973	0.000	4.743	0.000	0.000	NA	NA
Group 4	Mean	5.37	1.02	8.90	82.50	0.00	3.50	0.40	0.00	NA	NA
Group 4	SD	0.699	0.003	0.316	149.560	0.000	4.743	1.265	0.000	NA	NA

TREATMENT (WEEK 13)		ERY	LEU	EPI	RBC	PUS	CAST	CRY
TREATMENT (WEEK 13)		/µL	/µL	/field	/field	/field	/field	/field
Group 1	Mean	0.00	92.50	1.00	0.00	0.00	0.00	2.70
Group 1	SD	0.000	148.160	0.000	0.000	0.000	0.000	0.823
Group 2	Mean	0.00	132.50	1.00	0.00	0.00	0.00	2.50
Group 2	SD	0.000	196.515	0.000	0.000	0.000	0.000	0.850
Group 3	Mean	0.00	50.00	0.90	0.00	0.00	0.00	2.60
Group 3	SD	0.000	44.096	0.316	0.000	0.000	0.000	1.075
Group 4	Mean	2.50	92.50	1.00	0.00	0.00	0.00	2.40
Group 4	SD	7.906	148.160	0.000	0.000	0.000	0.000	1.075

Key: NA = Not Applicable

FEMALES

RECOVERY (Week 17)		Volume	SG	pH	Pro	GLU	KET	UBG	BIL	Color	Clarity
RECOVERY (Week 17)		mL	-	-	mg/dl	-	mg/dl	mg/dl	mg/dl	-	-
Group 1R	Mean	5.13	1.02	8.40	72.50	0.00	2.50	0.00	0.00	NA	NA
Group 1R	SD	0.457	0.002	0.699	151.589	0.000	2.635	0.000	0.000	NA	NA
Group 4R	Mean	5.25	1.02	8.40	162.50	0.00	5.00	0.10	0.00	NA	NA
Group 4R	SD	0.703	0.003	0.699	186.059	0.000	5.774	0.316	0.000	NA	NA

RECOVERY (Week 17)		ERY	LEU	EPI	RBC	PUS	CAST	CRY
RECOVERY (Week 17)		/µL	/µL	/field	/field	/field	/field	/field
Group 1R	Mean	5.00	47.50	0.80	0.00	0.00	0.00	2.60
Group 1R	SD	10.541	36.228	0.422	0.000	0.000	0.000	1.506
Group 4R	Mean	0.00	110.00	0.90	0.00	0.00	0.00	1.80
Group 4R	SD	0.000	142.009	0.568	0.000	0.000	0.000	0.632

Key: NA = Not Applicable

ORGAN WEIGHTS (G)

MALES

Organs		TREATMENT				RECOVERY
Organs		Group 1	Group 2	Group 3	Group 4	Group 1R	Group 4R
Body Weight (G)	Mean	369.47	360.84	364.16	355.55	394.22	396.05
	SD	37.01	32.47	28.26	42.83	29.22	29.14
	N	10	10	10	10	10	10
Adrenals	Mean	0.0882	0.1584	0.0904	0.0916	0.0879	0.0886
	SD	0.0262	0.2056	0.0220	0.0413	0.0181	0.0177
	N	10	10	10	10	10	10
Kidneys	Mean	2.0028	1.9861	2.0283	2.0210	2.0407	2.0053
	SD	0.2367	0.1940	0.1235	0.2424	0.1787	0.1864
	N	10	10	10	10	10	10
Liver	Mean	10.0878	10.0878	10.3265	10.4688	10.8801	11.3502
	SD	1.2526	1.2525	0.9455	1.4035	1.2492	0.8907
	N	10	10	10	10	10	10
Heart	Mean	1.1699	1.1751	1.1510	1.2151	1.2401	1.1954
	SD	0.1596	0.1235	0.1039	0.1373	0.1623	0.1102
	N	10	10	10	10	10	10
Thymus	Mean	0.4725	0.4662	0.4653	0.4646	0.4347	0.4638
	SD	0.0962	0.0700	0.0652	0.1016	0.0933	0.0683
	N	10	10	10	10	10	10

Organs		TREATMENT				RECOVERY
Organs		Group 1	Group 2	Group 3	Group 4	Group 1R	Group 4R
Spleen	Mean	0.7067	0.6786	0.6670	0.6829	0.6731	0.7299
	SD	0.1865	0.1228	0.0676	0.1051	0.0978	0.0997
	N	10	10	10	10	10	10
Testes	Mean	3.9717	3.9355	3.9715	3.9018	3.7815	4.0577
	SD	0.2906	0.2662	0.2892	0.3083	0.3458	0.2464
	N	10	10	10	10	10	10
Epididymides	Mean	1.6972	1.6259	1.6547	1.6242	1.6922	1.7497
	SD	0.1265	0.1590	0.1632	0.1931	0.1807	0.1652
	N	10	10	10	10	10	10
Prostate + Seminal vesicles with coagulating gland	Mean	2.8477	2.7193	2.8320	2.5825	3.0185	3.1790
	SD	0.2176	0.3672	0.4523	0.2898	0.4887	0.4975
	N	10	10	10	10	10	10
Brain	Mean	2.1798	2.1177	2.1717	2.1813	2.1824	2.1123
	SD	0.0894	0.0818	0.1193	0.0941	0.0653	0.1032
	N	10	10	10	10	10	10

FEMALES

Organs		TREATMENT				RECOVERY
Organs		Group 1	Group 2	Group 3	Group 4	Group 1R	Group 4R
Body Weight (G)	Mean	214.8	216.2	215.0	202.0	223.2	214.8
	SD	11.33	19.30	16.90	11.44	11.59	15.96
	N	10	10	10	10	10	10
Adrenals	Mean	0.1049	0.1012	0.1006	0.1022	0.1106	0.1084
	SD	0.0306	0.0397	0.0297	0.0278	0.0147	0.0213
	N	10	10	10	10	10	10
Kidneys	Mean	1.5102	1.4336	1.4582	1.3354	1.4855	1.3923
	SD	0.1102	0.1435	0.1783	0.1618	0.1164	0.1493
	N	10	10	10	10	10	10
Liver	Mean	7.0277	7.0319	7.2701	6.6108	6.9474	6.8568
	SD	0.5982	0.8065	0.9244	0.6169	0.8519	0.8777
	N	10	10	10	10	10	10
Heart	Mean	0.8961	0.8668	0.8401	0.8359	0.8643	0.9149
	SD	0.0950	0.0887	0.1118	0.0798	0.0933	0.2533
	N	10	10	10	10	10	10

Organs		TREATMENT				RECOVERY
Organs		Group 1	Group 2	Group 3	Group 4	Group 1R	Group 4R
Thymus	Mean	0.3618	0.3923	0.3553	0.3340	0.3215	0.2984
	SD	0.0683	0.0499	0.0726	0.0443	0.0645	0.0789
	N	10	10	10	10	10	10
Spleen	Mean	0.4599	0.4720	0.4976	0.4787	0.5188	0.4749
	SD	0.1710	0.0721	0.0668	0.0735	0.0700	0.0737
	N	10	10	10	10	10	10
Ovaries	Mean	0.1880	0.2528	0.1864	0.1662	0.1867	0.1719
	SD	0.0441	0.1440	0.0315	0.0378	0.0399	0.0465
	N	10	10	10	10	10	10
Uterus	Mean	0.7340	0.9319	0.7671	0.8519	0.7735	0.7830
	SD	0.0471	0.3839	0.2269	0.3005	0.1508	0.1130
	N	10	10	10	10	10	10
Brain	Mean	2.0723	2.0588	2.0869	1.9738	2.0267	2.0657
	SD	0.1224	0.1395	0.1025	0.1207	0.0893	0.1561
	N	10	10	10	10	10	10

1.1 ORGAN WEIGHT RATIO (%)

MALES

Organs		TREATMENT				RECOVERY
Organs		Group 1	Group 2	Group 3	Group 4	Group 1R	Group 4R
Body Weight (G)	Mean	369.47	360.84	364.16	355.55	394.22	396.05
	SD	37.01	32.47	28.26	42.83	29.22	29.14
	N	10	10	10	10	10	10
Adrenals	Mean	0.0246	0.0253	0.0249	0.0269	0.0225	0.0225
	SD	0.0097	0.0050	0.0059	0.0144	0.0053	0.0050
	N	10	10	10	10	10	10
Kidneys	Mean	0.5423	0.5507	0.5582	0.5694	0.5185	0.5066
	SD	0.0385	0.0284	0.0302	0.0354	0.0415	0.0359
	N	10	10	10	10	10	10
Liver	Mean	2.7925	2.7926	2.8364	2.9438*	2.7641	2.8677
	SD	0.1806	0.1806	0.1589	0.1594	0.2920	0.1270
	N	10	10	10	10	10	10
Heart	Mean	0.3164	0.3262	0.3161	0.3408	0.3147	0.3019
	SD	0.0243	0.0281	0.0149	0.0264	0.0341	0.0181
	N	10	10	10	10	10	10
Thymus	Mean	0.1276	0.1292	0.1285	0.1308	0.1103	0.1174
	SD	0.0213	0.0151	0.0200	0.0243	0.0237	0.0175
	N	10	10	10	10	10	10

*Significant at p ≤ 0.05 with Group 1

Organs		TREATMENT				RECOVERY
Organs		Group 1	Group 2	Group 3	Group 4	Group 1R	Group 4R
Spleen	Mean	0.1913	0.1871	0.1833	0.1922	0.1713	0.1848
	SD	0.0465	0.0192	0.0129	0.0211	0.0250	0.0244
	N	10	10	10	10	10	10
Testes	Mean	1.0800	1.0951	1.0939	1.1063	0.9610	1.0271
	SD	0.0786	0.0854	0.0894	0.1096	0.0788	0.0620
	N	10	10	10	10	10	10
Epididymides	Mean	0.4620	0.4531	0.4555	0.4597	0.4301	0.4438
	SD	0.0421	0.0555	0.0441	0.0536	0.0428	0.0512
	N	10	10	10	10	10	10
Prostate + Seminal vesicles with coagulating gland	Mean	0.7749	0.7536	0.7794	0.7332	0.7727	0.8093
	SD	0.0672	0.0714	0.1166	0.0972	0.1584	0.1557
	N	10	10	10	10	10	10
Brain	Mean	0.5955	0.5905	0.5986	0.6207	0.5565	0.5359
	SD	0.0657	0.0494	0.0444	0.0699	0.0469	0.0450
	N	10	10	10	10	10	10

FEMALES

Organs		TREATMENT				RECOVERY
Organs		Group 1	Group 2	Group 3	Group 4	Group 1R	Group 4R
Body Weight (G)	Mean	214.8	216.2	215.0	202.0	223.2	214.8
	SD	11.33	19.30	16.90	11.44	11.59	15.96
	N	10	10	10	10	10	10
Adrenals	Mean	0.0492	0.0473	0.0472	0.0511	0.0498	0.0505
	SD	0.0156	0.0204	0.0144	0.0154	0.0081	0.0098
	N	10	10	10	10	10	10
Kidneys	Mean	0.7046	0.6634	0.6778	0.6598	0.6667	0.6487
	SD	0.0585	0.0378	0.0618	0.0535	0.0560	0.0561
	N	10	10	10	10	10	10
Liver	Mean	3.2733	3.2555	3.3861	3.2715	3.1076	3.1889
	SD	0.2270	0.2767	0.4064	0.2250	0.2989	0.2723
	N	10	10	10	10	10	10
Heart	Mean	0.4182	0.4018	0.3904	0.4139	0.3873	0.4273
	SD	0.0483	0.0358	0.0380	0.0340	0.0366	0.1243
	N	10	10	10	10	10	10

Organs		TREATMENT				RECOVERY
Organs		Group 1	Group 2	Group 3	Group 4	Group 1R	Group 4R
Thymus	Mean	0.1681	0.1832	0.1648	0.1649	0.1447	0.1396
	SD	0.0275	0.0314	0.0291	0.0155	0.0320	0.0389
	N	10	10	10	10	10	10
Spleen	Mean	0.2147	0.2186	0.2315	0.2366	0.2321	0.2209
	SD	0.0797	0.0293	0.0268	0.0296	0.0247	0.0283
	N	10	10	10	10	10	10
Ovaries	Mean	0.0881	0.1176	0.0876	0.0829	0.0831	0.0802
	SD	0.0228	0.0680	0.0184	0.0215	0.0170	0.0226
	N	10	10	10	10	10	10
Uterus	Mean	0.3433	0.4333	0.3587	0.4170	0.3486	0.3646
	SD	0.0371	0.1807	0.1134	0.1243	0.0769	0.0463
	N	10	10	10	10	10	10
Brain	Mean	0.9689	0.9570	0.9736	0.9801	0.9104	0.9659
	SD	0.0975	0.0852	0.0526	0.0857	0.0642	0.0943
	N	10	10	10	10	10	10

Conclusions:

In conclusion, under the conditions of this experiment the No Observed Adverse Effect Level (NOAEL) was 400 mg/kg body weight.

Executive summary:

The test item Hordaphos MDB was administered daily by oral route to Wistar rats for a period of 90 consecutive days. A recovery period of 28 days without test item treatment was included in this study.

Three groups consisting of 10 male and 10 female rats each were treated at dose levels of 50, 200 and 400 mg/kg, respectively.Concurrently, vehicle control, vehicle control recovery and test item high dose recovery groups (400 mg/kg) were used in the present study.

All the animals survived up to the scheduled sacrifice.

No test item related changes in clinical signs, detailed clinical observation, functional observation battery, ophthalmic examination findings, organ weight and body weight changes were noted. The dullness observed in the in high dose (G4 & G4R) were considered to be without toxicological relevance.

Minor changes in hematological, clinical biochemistry, body weight, feed consumption and urine analysis were recorded at the end of treatment and recovery periods, but the variations were considered incidental as there was no dose dependent trend in the observed variations and the respective tissues revealed no abnormalities in macroscopic and histopathological examination.

Post-mortem examinations revealed no test item-related macroscopic findings in treated animals.

Microscopic examination revealed abnormalities in non glandular stomach attributable to the test item HORDAPHOS MDB at 50, 200 and 400 mg/kg bw in both sexes. The lesion was considered non adverse as complete recovery was observed at the end of 28 day recovery period.